FREEZING TECHNOLOGY

Preservation of Fish by Chilling and Freezing Technology

This course deals with the preservation of sea foods by chilling and freezing techniques.
As we know, fish is a perishable commodity, its mainly because of the composition. Nearly 70%
of the fish body constitutes water and 30% consist of protein, fat, minerals, and vitamin and
non-protein nitrogen extractives. At ambient temperature fish muscle undergo rapid biochemical
changes and creates a favorable environment for microorganisms to grow. This in turn
responsible for the production of fouls smell and makes muscle spoiled and unsuitable for human
consumption. Fish contains all essential amino acids (EAA) that is necessary for human body.
Poly unsaturated a fatty acid (PUFA) (Omega-3-fatty acids) of fish is good for human health.
Therefore it is imperative to preserve the fish immediately after catch to get the EAA & PUFA
without any deterioration. How it can be preserved?. It is possible by lowering the temperature
by direct (icing) or indirect (chill storage) methods. The main principle of chilling by ice is, it
lowers the temperature of fish body from 30°C to 5°C. This greatly affect the mesophilic
bacterial flora of fish and its growth is completely arrested by lowering of temperature and also
slows down the biochemical activity, there by preserving quality to the extended time. Various
methods of chilling is explained in detail.
To consume the fish as similar to fresh fish, it has to be subjected to freezing process.
Different methods of freezing fish, equipments used, refrigeration load and refrigerants used are
discussed with illustration in detail.
Before consuming, frozen fish has to be thawed to obtain fish in normal form. Various
methods of thawing including latest techniques are described.

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 Normally frozen fish will be stored in cold store where the temperature is maintained at
-18°C. During storage due to various factors frozen fish undergo physical & chemical changes.
Quality changes during storage, its preservative methods are explained cleanly storage condition
is very important for best quality sea food. Therefore, the correct layout of cold store with all
factors, mobile store (containers) was also described.
Packaging of food is important in maintaining the wholesomeness of the product. Various
types of packaging materials and their requirements were elaborated.
To export the sea food it is essential to register the processing unit to European Union
under their law. Therefore every approved unit must have a HACCP plan for that particular firm.
Before preparing a plan one must know the classification of physical, chemical and biological
hazards including Bio-toxin. All the above hazards are discussed in detail. Extension of shelf life
by vacuum packing & Modified atmospheric packaging (MAP) were also documented.
By undergoing this course one must learns the preservation technique by freezing and
how to main the hygienic condition of the fish and export formalities, rules and regulations of
importing countries.

Unit 1: Introduction to fish muscle function

Chapter 1: Introduction and historical developments in low temperature preservation of

fish

1.1.1.Introduction

Several food preservation techniques were evolved in order to enable man to use food
produced at various places from age old times. Such methods of preservation of food are sun
drying, salting, keeping food in ice, smoking etc.

Fish is a highly perishable commodity as it contains good amount of nutrients and hence
there is a greater need to preserve it. Without preservation it is impossible to transport and
distribute fish to interior places in good condition.

1.1.1.1. Historical developments in low temperature preservation of foods

In olden days, in places which were covered with snow or ice, the flesh of hunted animals
and fish were covered with ice thereby fish was frozen and quality is preserved. However,
freezing of fish/ foods by artificial methods is a recent development.

Towards the end of 19th century compressors were developed and because of this
machine, novel methods of fish freezing were developed. First patent on freezing of fish using
cold air was given to V.Benzamine of England in 1842. In the year 1861, similar patent was
awarded to E. Piper of U.S.A. Who used salt ice mixture to freeze food. Russians were pioneers

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in freezing fish by using machines. They used compressors on board the fishing vessel to freeze
fish and also built a freezing plant (factory) in a place called Astrakhan. In 1898 immersion
freezing system was developed for freezing fish. Nikhoidahl, a scientist from Norway developed
spray freezing method. In 1925, a technician named Cook developed contact plate freezer which
was further improved by Karensbirdseye in 1929. Similarly methods of freezing and packing it
in consumer packs were developed subsequently.

1.1.1.2. Demand for low temperature preserved fish

In most of the processing methods such as salting, drying, canning etc., changes in the
sensory characteristics take place. Most people like fish with characteristics as existing in freshly
caught fish. Therefore, the principal problem encountered by fishing industry is to preserve fish
in such a way that it does not lose its original sensory characteristics. Even for processing by
other methods, initial preservation of fish is done by low temperature preservation technique
such as icing. Hence low temperature preservation of fish has become increasingly popular
in recent years.

Chapter 2: Structure and function of fish muscle

1.2.1. Introduction

Most of the fish muscle tissue is white but, depending on the species, many fish will have a
certain amount of dark tissue of a brown or reddish colour located just under the skin along the
side of the body. In pelagic fish, i.e., species such as herring and mackerel which swim more or
less continuously, up to 48% of the body weight may consist of dark muscle (Love, 1970). In
demersal fish, i.e., species which feed on the bottom and only move periodically, the amount of
dark muscle is very small.From technology point of view, the high lipid content of dark muscle is
important because of problems with rancidity.

The energy source for ATP generation in the light muscle is glycogen, whereas the dark
muscle may use lipids. A major difference is, that the dark muscle contains much more
mitochondria than light muscle, thus enabling the dark muscle to operate an extensive aerobic
energy metabolism resulting in CO2 and H2O as the end products. The light muscle, mostly
generating energy by the anaerobic metabolism, accumulates as lactic acid which has to be
transported to the liver for further metabolization. In addition, the dark muscle is reported to
possess functions similar to those are found in the liver.

1.2.1.1. Lipids

The lipids present in teleost fish species can be divided into two major groups: the
phospholipids and the triglycerides. The phospholipids make up the integral structure of the unit

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membranes in the cells; thus, they are often called structural lipids. The triglycerides are lipids
used for storage of energy in fat depots, usually within special fat cells surrounded by a
phospholipid membrane and a weak collagen network. The triglycerides are often termed depot
fat. A few fish have wax esters as part of their depot fats.

The mobilization of energy is much faster in light muscle than in dark muscle, but the
formation of lactic acid creates fatigue, leaving the muscle unable to work for long periods at
maximum speed. Thus, the dark muscle is used for continuous swimming activities and the light
muscle for quick bursts, such as when the fish is about to catch a prey or to escape a predator.

In elasmobranchs, such as sharks, a significant quantity of the lipid is stored in the liver
and may consist of fats like diacyl-alkyl-glyceryl esters or squalene. Some sharks may have
liver oils with a minimum of 80% of the lipid as unsaponifiable substance, mostly in the form of
squalene.

Fish lipids differ from mammalian lipids. The main difference is that fish lipids include up
to 40% of long-chain fatty acids (14-22 carbon atoms) which are highly unsaturated. Mammalian
fat will rarely contain more than two double bonds per fatty acid molecule while the depot fats of
fish contain several fatty acids with five or six double bonds.

2.1.2. Proteins

The proteins in fish muscle tissue can be divided into the following three groups:

1. Structural proteins (actin, myosin, tropomyosin and actomyosin), which constitute

70-80% of the total protein content (compared with 40% in mammals). These proteins are
soluble in neutral salt solutions of fairly high ionic strength (> 0.5M).
2. Sarcoplasmic proteins (myoalbumin, globulin and enzymes) which are soluble in
neutral salt solutions of low ionic strength (<0.15M). This fraction constitutes 25-30% of
the protein.
3. Connective tissue proteins (collagen), which constitute approximately 3% of the protein
in teleost and about 10% in elasmobranch (compared with 7% in mammals).

The majority of the sarcoplasmic proteins are enzymes participate in the cell metabolism,
such as the anaerobic energy conversion from glycogen to ATP. If the organelles with in the
muscle cells are broken, this protein fraction may also contain the metabolic enzymes localized
inside the endoplasmic reticulum, mitochondria and lysosomes. The fact that the composition
of the sarcoplasmic protein fraction changes when the organelles are broken was suggested
as a method for differentiating fresh from frozen fish, under the assumption that the
organelles were intact until freezing. The proteins in the sarcoplasmic fraction are
excellently suited to distinguish between different fish species, as all the different species
have their characteristic band pattern, when separated by the isoelectric focusing method. Fish
protein is an excellent source of all essential amino-acids. In diets based mainly on cereals, a
supplement of fish can raise the biological value significantly.

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 1.2.1.3. N – containing extractives

The N-containing extractives can be defined as the water-soluble, low molecular

weight, nitrogen-containing compounds of non-protein nature. This NPN-fraction (non-protein
nitrogen) constitutes from 9 to 18% of the total nitrogen in teleosts. The major components
in this fraction are: volatile bases such as ammonia and trimethylamine oxide (TMAO), creatine,
free amino-acids, nucleotides and purine bases, and urea in the case of cartilaginous fishes.

Fish Crustacean oultry Mammalian

Compound in mg/100g s muscle
wet weight Cod Herrin Shark Lobster Leg
g specie muscl
s e
1) Total extractives 1200 1200 3000 5500 1200 3500
2) Total free amino – acids 75 300 100 3000 440 350
Arginine <10 <10 <10 750 <20 <10
Glycine 20 20 20 100-1000 <20 <10
Glutamic acid <10 <10 <10 270 55 36
Histidine <1.0 86 <1.0 -- <10 <10
Proline <1.0 <1.0 <1.0 750 <10 <10
3) Creatine 400 400 300 0 -- 550
4) Betaine 0 0 150 100 -- --
5)Trimethylamine oxide 350 250 500-10 100 0 0
00

The amount of TMAO in the muscle tissue depends on the species, season, fishing
ground, etc. In general, the highest amount is found in elasmobranchs and squid (75-250 mg
N/100g) followed by cod (60-120mg N/100g),while flatfish and pelagic fish have the least.
Pelagic fish (sardines, tuna, and mackerel) have their highest concentration of TMAO in the dark
muscle while demersal, white fleshed fish have a much higher content in the white muscle.
Quantitatively, the main component of the NPN-fraction is creatine. In resting fish, most of the
creatine is phosphorylated and supplies energy for muscular contraction.

1.2.1.4. Vitamins and Minerals

In general, fish meat is a good source of vitamin B and in the case of fatty species A and D
vitamins. Some freshwater species such as carp have high thiaminase activity so the thiamine
content in these species is usually low. Fish meat is regarded as a valuable source of calcium and
phosphorus in particular iron, copper and selenium in general. Saltwater fish have a high
content of iodine.

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 Some Mineral constituents of fish muscle

Elements Content (mg/100 g)

Sodium 30-134

Potassium 19-502

Calcium 19-881

Magnesium 4.5-452

Phosphorus 68-550

Iron 1-5.6

Chlorine 3-761

Iodine 0-2.73

Chapter 3: Postmortem changes in fish

1.3.1 Changes in raw fresh fish

Immediately after death the muscle is totally relaxed and the elastic texture usually
persists for some hours, thereafter the muscle will contract. When it becomes hard and stiffs the
whole body becomes inflexible and the fish is in rigor mortis.The rate in onset and resolution of
rigor varies from species to species and is affected by temperature, handling, size and physical
condition of the fish. Rigor mortis starts immediately or shortly after death, if the fish is starved
and the glycogen reserves are depleted, or if the fish is stressed during fishing.

In rigor the fish body will be completely stiff; the filleting yield will be very poor,
and rough handling can cause gaping. If the fillets are made from pre-rigor muscle, it contract
freely and the fillets will shorten following the onset of rigor. Dark muscle may shrink up to 52%
and white muscle up to 15% of the original length (Buttkus, 1963). If the pre-rigor fish is cooked
the texture will be very soft and pasty. In contrast, the texture is tough but not dry when the fish
is cooked in rigor. Post-rigor the flesh will become firm, succulent, elastic and best suited for
processing.

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 1.3.2 Changes in eating quality

Changes in the eating quality of chilled fish during storage can be assessed by daily
organoleptic examination of the cooked flesh.

Phase 1: The fish is very fresh and has a sweet, sea weedy and delicate taste. The taste
can be very slightly metallic. In cod, haddock, whiting and flounder, the sweet taste is
maximized 2-3 days after catching.

Phase 2: There is a loss of the characteristic odour and taste. The flesh becomes neutral
but has no off-flavours. The texture is still pleasant.

Phase 3: There is sign of spoilage and a range of volatile, unpleasant-smelling

substances are produced depending on the fish species and type of spoilage (aerobic, anaerobic).
One of the volatile compounds may be trimethylamine (TMA) derived from the bacterial
reduction of trimethylaminoxide (TMAO). TMA has a very characteristic “fishy” smell. At the
beginning of the phase the off-flavour may be slightly sour, fruity and slightly bitter, especially
in fatty fish. During the latter stages sickly sweet, cabbage-like, ammoniacal, sulphurous and
rancid smells develop. The texture becomes either soft or watery of tough and dry.

Phase 4: The fish can be characterized as spoiled and putrid.

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8
 1.3.3 Autolytic changes

Autolysis means “self-digestion”. It has been known for many years that there are at least
two types of fish spoilage: enzymatic and bacterial.

Muscle enzymes and their activity: At the point of death, the supply of oxygen to the
muscle tissue is interrupted because the blood is no longer pumped by the heart and is not
circulated through the gills. Since no oxygen is available for normal respiration, the production
of energy from ingested nutrients is greatly restricted. Figure illustrates the normal pathway for
the production of muscle energy in most living teleost fish (bony finfish). Glycogen (stored
carbohydrate) or fat is oxidized or “burned” by the tissue enzymes in a series of reactions which
ultimately produce carbon dioxide (CO2), water and the energy-rich organic compound
adenosine triphosphate (ATP). This type of respiration takes place in two stages: an anaerobic
and an aerobic stage. The latter depends on the continued presence of oxygen (O2) which is
only available from the circulatory system. Most crustaceans are capable of respiring outside the
aquatic environment by absorption of atmospheric oxygen for limited period.

1.3.3.1 Cytoplasm anaerobic metabolism

Aerobic and anaerobic breakdown of glycogen in vertebrate & invertebrate muscle

tissue.

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 Under anaerobic conditions, ATP may be synthesized by two other important pathways
from creatine phosphate or from arginine phosphate. The former source of energy is restricted to
vertebrate muscle (teleost fish) while the latter is characteristic of some invertebrates such as the
cephalopods (squid and octopus). In either case, ATP production ceases when the creatine or
arginine phosphates are depleted. It is interesting to note that octopine is the end-product from
the anaerobic metabolism of cephalopods and is not acidic (unlike lactate), thus any changes in
post mortem pH in such animals are not related to the production of lactic acid from glycogen.
For most teleost fish, glycolysis is the only possible pathway for the production of energy once
the heart stops beating.

After death, when the regeneration ceases, the ATP is rapidly degraded i.e. after death,
the anaerobic muscle cannot maintain its normal level of ATP, and when the intracellular ATP
level declines from 7 - 10 µmoles/g to <1.0 µmoles/g tissue, the muscle enters rigor mortis. Post
mortem glycolysis results in the accumulation of lactic acid which in turn lowers the pH of the
muscle. In cod, the pH drops from 6.8 to an ultimate pH of 6.1- 6.5. In some species of fish, the
final pH may be lower: in large mackerel, the ultimate rigor pH may be as low as 5.8 - 6.0 and as
low as 5.4 - 5.6 in tuna and halibut, however such low pH levels are unusual in marine teleosts.
In general, fish muscle contains a relatively low level of glycogen compared to mammals, thus
far less lactic acid is generated after death. Also, the nutritional status of the fish and the amount
of stress and exercise encountered before death will have a significant effect on the levels of
stored glycogen and consequently on the ultimate post mortem pH. As a rule, well-rested,
well-fed fish contain more glycogen than exhausted fish. It is show that bleeding of fish
significantly reduced the post mortem production of lactic acid.

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 The post mortem reduction in pH of fish muscle has an effect on the physical properties
of the muscle. As the pH drops, the net surface charge on the muscle proteins is reduced, causing
them to partially denature and lose some of their water-holding capacity. Muscle tissue in the
state of rigor mortis loses its moisture when cooked and is particularly unsuitable for further
processing which involves heating, since heat denaturation enhances the water loss. Loss of
water has a detrimental effect on the texture of fish muscle and it has been shown by Love
(1975) that there is an inverse relationship between muscle toughness and pH, unacceptable
levels of toughness (and water-loss on cooking) occurs at lower pH levels.

1.3.3.2 Autolysis and nucleotide catabolism

Autolysis and nucleotide catabolism

According to the present understanding, rigor mortis is caused by bonding of the

myosin heads, extending radially from the thick microfibrils, to the active centers in actin units
of the thin filaments. This leads to the formation of a rigid structure of interconnected
myofilaments. This reaction between these major muscle proteins is made possible by changes in
the regulatory proteins. These changes are in turn induced by an increase in the concentration of
Ca2+ in the sarcoplasm. In a resting muscle, the concentration of free Ca2+ in the sarcoplasm is
lower than 10-7M. This level is maintained because of the action of various calcium pumps,
located in the cell membrane and in the sarcoplasmic reticulum, which drives the Ca2+ out of the
sarcoplasm, against the concentration gradient. The calcium pumps work at the expense of
energy from ATP hydrolysis. After the death of the animal, a gradual depletion of ATP in the
muscles takes place, due to exhaustion of the creatine phosphate and glycogen reserves. This
impairs the action of the calcium pumps and increases the concentration of Ca2+ in the
sarcoplasm. At ATP concentrations below 10-4M and that of Ca2+ above 10-6M, rigor mortis sets
in. As the individual muscle fibers contain different quantities of ATP, they do not enter rigor at
the same time. Thus, stiffness in a muscle sets in gradually.

The resolution of rigor is a process still not completely understood but always results
in the subsequent softening (relaxation) of the muscle tissue and is thought to be related to the
activation of one or more of the naturally-occurring muscle enzymes, digesting away certain
components of the rigor mortis complex. The softening of the muscle during resolution of
rigor (and eventually spoilage processes) is coincidental with the autolytic changes. Among
the changes, one of the first to be recognized was the degradation of ATP-related compounds in a
more-or-less predictable manner after death.

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 Degradation of ATP to hypoxanthine (Hx) and ribose
Freshness is expressed by K-value. This K-value expresses the relationship between
inosine and hypoxanthine and the total amount of ATP-related compounds:
HxR + Hx
K(%) = ------------------------------------------------------------------------- x 100
ATP + ADP + AMP + IMP + HxR + Hx

Very fresh fish, have low K-values. K-values increase gradually which is species
dependent. Inosine is said to be more or less flavourless, while hypoxanthine has been reported
to impart a bitter flavour in spoiling fish (Spinelli, 1965). Hx is considered to have a direct
effect on the perceived bitter off-flavour of spoiled fish (Hughes and Jones, 1966). It is now
widely accepted that IMP is responsible for the desirable fresh fish flavour which is only
present in top quality seafood.

Chapter 4: Bacteriological changes

1.4.1 Native bacterial flora of fishes

The flesh and body fluids of live healthy fish are generally free from bacteria, i.e. sterile.
But, even when the fish is alive, they harbour bacteria, mainly on three sites of their body (i.e.)
the slime on the skin surface the gill tissue and the intestine. Bacteria, which are naturally
present on fishes, are called the native bacterial flora of fish. The population and nature of
such flora depend on the waters from where the fish were caught, i.e. whether seawater, brackish
water or fresh water. Seawater has more dissolved salts, i.e. a higher salinity than both brackish
and fresh water.
Salinity range of water bodies

Water Salinity range

Sea water 30-36 ppt

Brackish water 10-28 ppt

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 Fresh water 0.2-1.0 ppt

Depending on the salinity and pollution range of waters, the bacterial flora of fishes varies from
water to water. Generally the bacterial population of the fishes from tropical water is in the
following ranges .
Bacterial population of marine fishes from tropical waters

Sample Oil sardine (Sardinella Indian mackerel

longiceps) (Rastrelliger kanagurta)

Skin with slime/cm2 103-107 104-106

Gills/g 105-108 104-109

Intestine with contents/g 105-109 105-108

Bacterial population of brackish water fishes

Sample Pearl spot Milkfish

(Etroplus suratensis ) (Chanos chanos)

Skin with slime/cm2 103-104 103-105

Gills/g 104-108 105-108

Intestine with contents/g 105-108 106-108

Bacterial population of fresh water fishes

Sample Mrigal (Cirrhinus

Rohu (Labeo rohita) mrigala)

Skin with slime/cm2 104-105 104-105

Gills/g 104-106 105-107

Intestine with 105-107 104-106

contents/g

Generally, the bacterial populations on the skin surface are the least, while bacterial
counts in the intestine are the highest. Bacterial counts in the gill tissue are more or less
between the bacterial counts of skin surface and the intestine. Bacterial populations exhibit
seasonal variations. During warmer months, the counts used to be higher while during colder
seasons, the bacterial counts are lower. Qualitatively, the composition of bacterial flora of the
fishes from the three waters differ considerably. Majority of the bacterial flora of marine
fishes are gram negative, non-spore forming (asporogenous) rods or cocci. Bacterial flora of
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fresh water fishes is predominantly gram positive in nature. The microflora of the brackish
water fishes are evenly composed of both gram +ves and gram –ves.
From these examples, it can be seen that in the case of marine fishes, not only there is a
larger proportion of gram-negative bacteria, but also, there is a large number of bacterial types.
But for fresh water fishes, the number of bacterial genera is quite limited.
a. Microbial flora on the skin surface of marine fish:
Gram negative: Vibrio , Pseudomonas , Moraxella , Acinetobacter , Flavobacteria , Aeromonas ,
Photobacteria.
Gram positive: Arthrobacter , Micrococcus , Bacillus.
b. Bacterial flora on the skin surface of brackish water fish:
Gram negative: Pseudomonas , Alcaligenes , Flavobacter , Moraxella , Vibrio , Acinetobacter ,
Coliforms.
Gram positive: Micrococcus , Bacillus , Arthrobacter , Lactobacillus , Streptococci.
c. Bacterial flora on the skin surface of freshwater fish:
Gram negative: Acinetobacter ,Pseudomonas , Coliforms.
Grampositive: Micrococcus , Bacillus.

1.4.2 Factors that influence the growth of microorganisms

There are a number of external and internal factors (intrinsic to the substrate, that is, the
foods in which they grow) that affect the microbial growth.
Of the different external factors, temperature is the most important one.
(1) Thermophiles (high-temperature loving kind): growth optimum above
45 o C (35 o to 65 o C).
(2) Mesophiles (medium- temperature loving kind): growth optimum above
25o to 37 o C (15 o to 45 o C).
3) Psychrotrophs (cold-temperature tolerant kind): growth temperature is between 0 o -
25 C (optimum 15 o - 25 oC).
o

The bacteria on temperate water fish are all classified according to their growth
temperature range as either psychrotrophs or psychrophiles. Psychrotrophs (cold-tolerant) are
bacteria capable of growth at 0oC but with optimum around 25oC. Psychrophiles
(cold-loving)are bacteria with maximum growth temperature around 20oC and optimum
temperature at 15oC (Morita, 1975). In warmer waters, higher numbers of mesophiles can be
isolated. The microflora on temperate water fish is dominated by psychrotrophic gram-negative
rodshaped bacteria belonging to the genera Pseudomonas, Moraxella, Acinetobacter, Shewanella
and Flavobacterium. Members of the Vibrionaceae (Vibrio and Photobacterium) and the

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Aeromonadaceae (Aeromonas spp.) are also common aquatic bacteria and typical of the fish
flora. Gram-positive organisms are Bacillus, Micrococcus, Clostridium, Lactobacillus and
Coryneforms can also be found in varying proportions, but in general, Gram-negative bacteria
dominate the microflora.

Interaction between growth factors

1) Microbial growth is arrested even when one of the growth factor is controlled. For
example, bacterial growth slows down under reduced pH or acidic conditions.

(2) If more than one factor becomes limiting, microbial growth is drastically curtailed or
even stopped. For example bacterial growth would stop under acidic conditions combined with
low temperature and lower water activity(aw).

Chapter 5: Spoilage of fish

1.5.1 Spoilage of fish

Spoilage of fish, post mortem, is mainly due to (1) oxidation, (2) autolysis and (3)
bacteria. The major cause of spoilage of fish is bacteria, particularly in the case of marine
fishes.

The flesh and body fluids of newly caught fish are free from bacteria (except when the
fish has bacterial disease). The bacteria present on skin, adhering slime gills and intestine are
normally saprophytic. Once the fish is dead, these bacteria invade the fish tissue.There are three
main routes for this attack.

1. From the gills into the flesh through the vasculatory (circulatory) system.
2. Through the skin by penetration.
3. Through the peritoneal lining (from the intestinal cavity).

Invation of bacteria through the first and second routes is faster. Entry through the
peritonial lining can take place only after perforation of stomach and intestinal walls, which
normally takes longer time.

The fish muscle contains 15 to 18% protein. Bacteria attack the protein and break it down
to peptides and amino acids. Initially, bacteria live and multiply in the fish tissue, utilizing the
low molecular weight compounds like carbohydrates and amino acids present in small quantities
in the muscle. Due to post-mortem enzymatic break down of the macromolecules in the muscle,
enough low molecular weight compounds are formed, which serve as the food of bacteria.
Subsequently bacteria elaborate proteolytic enzymes, which break down proteins to peptone,
polypeptides, lower peptides and finally to amino acids. Bacteria will metabolize amino acids, in

15
different ways leading to the production of odouriferous and foul smelling compounds like
ammonia, hydrogen sulphide, mercaptans, indole, amines and organic acids.

When the fish is left in ambient temperature, which is usually 28 ±4°C, tropical fishes get
spoiled within 6 to 12 hrs. depending on their size. In order to prevent such spoilage, many
methods are in practice. Drying, icing, freezing and use of chemicals are some of the usual
methods. The basic principle involved in these methods of preservation of fish is to control the
activities of the microorganisms.

Typical spoilage compounds during spoilage of fresh fish stored aerobically or

packed in ice or at ambient temperature

Specific spoilage organism Typical spoilage compounds

Shewanella putrefaeciens TMA, H2S, CH3SH, (CH3)2S, Hx

Photobacterium phosphoreum TMA, H2S

Pseudomonas spp.
Ketones, aldehydes, esters, non-H2S
Sulphides
Vibrionaceae TMA, H2S

Anaerobic spoilers NH3, acetic, butyric and propionic acid

Substrate and off-odour / off-flavour compounds produced by bacteria during

spoilage of fish

Substrate Compounds produced by bacterial action

TMAO TMA

Cysteine H2S

Methionine CH3SH, (CH3)2S

Carbohydrates and lactate Acetate, CO2, H2O

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Inosine, IMP
Hypoxanthine
Amino-acids (glycine, serine, leucine) Esters, ketones, aldehydes

Amino-acids, urea NH3

17
Unit 2: Chilling of fish

Chapter 1: Fresh fish handling

2.1.1. Introduction to Icing

Icing is the most prevalent method of preserving fish. Ideal icing involves packing crushed
ice and fish alternatively in insulated boxes, in the fish to ice ratio of 1:1 (w/w). By this, the
temperature of the fish is lowered to near 1 to 2C in about 2-3 hrs (the melting of the ice needs
80 calories of heat/g and this heat is removed from the fish in contact with ice and hence, the fish
get cooled). This lowering of temperature brings about

1. arrest of almost all enzymatic changes,

2. killing of about 50-60% of the mesophilic bacteria and
3. slowing down of the activities and growth of all other bacteria, which are cold-loving
(psychrophilic) and cold-tolerant (psychrotrophic).

As a combined effect of all these three factors, the spoilage of fish is delayed to a
considerable length of time in ice. During iced storage of fish, there is an initial drop of bacterial
count due to the death of the cold sensitive mesophiles. The surviving cold tolerant bacteria,
however, get adapted to growth in low temperature. Consequently, there is a gradual increase in
population, which takes about 6 to 8 days to reach a count of one million per gram or above. By
that time, the fish has reached the stage of incipient spoilage.

Qualitatively, there is a selection of bacterial flora during iced storage of fish. Irrespective
of the composition of the initial flora, the Pseudomonas/ Alteromonas group emerges as the
predominant group of bacteria at the time of spoilage. This is because most of the
psychrotrophic bacteria capable of spoilage belong to these genera.

In the case of tropical fishes, it is not the psychrophiles, but the psychrotrophs, which are
the actual spoilers during iced storage. These psychrotrophs, whose population is very low
in the fresh tropical fish, easily adopt to grow at low temperature during iced storage and
grow very rapidly and spoil the fish. Further, psychrotrophs have a shorter generation time
compared with psychrophiles.

2.1.2. Advantages of chilling of fish with ice

i) Temperature reduction. It reduces the temperature to about 0oC. The growth of spoilage and
pathogenic micro-organisms is reduced, thus reducing the spoilage rate.
ii) Melting ice keeps fish moist. This action mainly prevents surface dehydration and reduces
weight losses. Melting water also increases the heat transport between fish and ice surfaces
(water conducts heat better than air): the quickest practical chilling rate is obtained in a slurry of
water and ice (e.g., the CSW system).
iii) Advantageous physical properties. Ice has some advantages when compared with other
cooling methods, including refrigeration by air. The properties can be listed as follows:

18
a) Ice has a large cooling capacity.The latent heat of fusion of ice is about 80 Kcal/kg. This
means that a comparatively small amount of ice will be needed to cool 1 kg of fish.
b) Ice melting is a self-contained temperature control system. Ice melting is a change in the
physical state of ice (from solid to liquid), and it occurs at a constant temperature (0oC).
iv) Convenience. Ice has a number of practical properties that makes its use advantageous. They
are:
a) It is a potable cooling method. It can be easily stored, transported and used. Depending on
the type of ice, it can be distributed uniformly around fish.
b) Raw material to produce ice is widely available. Although clean, pure water is becoming
increasingly difficult to find, it is still possible to consider it a widely available raw material
and it should be properly treated, e.g., chlorination.
c) Ice can be a relatively cheap method of preserving fish. This is particularly true if ice is
properly produced (avoiding wastage of energy at ice plant level), stored (to avoid losses)
and utilized properly (not wasted).
d) Ice is a safe food grade substance. If produced properly utilizing drinking water, ice is a safe
food substance and does not cause any harm either to consumers or those handling it. Ice
should be handled as food.
v) Extended shelf life.
The overall reason for icing fish is to extend shelf life fresh fish of in a relatively simple way
as compared to storage of un-iced fish at ambient temperatures above 0oC. Extension of shelf life
means producing safe fresh fish of acceptable quality.

2.1.3. Chilled storage

In general, two types of plastic fish boxes are used: stack-only and nest/stack boxes.

1. The handling rate is necessary to prevent quality loss because of delayed icing.
Pre-chilling can be of advantage to compensate lack in handling rate.
2. Handling methods, which make it possible to guarantee that the icing procedure is
sufficient to chill the fish to 0oC and maintain this temperature until landing.
3. The hold must be constructed in such a way, that safe and easy stacking of the boxes can
take place.
4. Hold insulation of a relatively high quality should be considered. A small mechanical
refrigeration plant can be of advantage. Air temperature in the hold should be +1o or -3oC.

Use of antimicrobial agents in ice and cold liquids

By ordinary method of icing, fish can be preserved for 10-12 days. However, studies have shown
that use of antimicrobial agents in ice or cold liquids, enhance the shelf life of fish by 60 to
100%. The following antimicrobial agents were used.

a) Chlorine compounds

If liquid chlorine or bleaching powder is mixed with ice or cold liquids, shelf life of fish can be
enhanced. If the cooling media (ice or chilled liquid) contains less than 30 ppm of chlorine, no
damage is done to fish. However if the proportion is more than this, the fish gets acrid smell.

19
Also due to oxidation, the colour of fish may change. By this method fish can be preserved for
18-20 days.

b)Antibiotics

By the use of antibiotics in ice or cold liquids, shelf life of chilled fish can be increased.
Among the antibiotics, chlorotetracycline and oxytetracycline are important. These are powerful
broad spectrum antibiotics and are very effective even at low concentrations. Antibiotics were
used for chilling fish as follows.

1. By dip treatment in antibiotic solution and then chilling.

2. Incorporating antibiotic in water and then ice is prepared and used for chilling.
3. Incorporating in chilling liquids (CSW or RSW)
4. It has been found that dip treatment of fish in water containing 30 to 100 mg of CTC for
1-5 min and then icing increases the shelf life by 5-10 days. Ice containing 5 mg/kg of
CTC, when used, increase the shelf life of fish from 10 days to 20-22 days. However at
present use of antibiotics in banned as per Codex Alimentarius standards.

2.1.4 Storage method

There are three methods of storing fish in ice on fishing vessels

Bulking:

The fish hold is a insulated chamber located at the near side of the Boat. It is divided
into different compartments using wooden boards supported by an upright beam. A layer of ice
at least 5 cm thick is spread over the bottom of a compartment followed by a layer of fish. Ice is
then spread over the fish and around the edges so that the fish are not in direct contact with the
sides of the board.

Further layers of fish and ice are added until a depth of about 45cm ice and fish is
achieved, with a layer of 5 cm of ice at the top. A horizontal wooden board is now placed over
the section. The wooden board must be supported by the stanchion structures, not by the fish and
ice in the lower compartment. More fish and ice are added in the same way, again to a depth of
45cm. The operation is repeated until the compartment is full. Wooden boards and stanchions
must be kept clean and out of direct contact with the fish.

Shelving: The fish hold is divided into sections as it is for bulking but this time removable
shelves spaced at about 23 cm are used for holding the fish. The lowest shelf is covered with a
layer of at least 5 cm of ice. Fish are placed in rows on the ice and more ice is used to cover the
fish to about 5 cm. Only one layer of fish is to be put on to each shelf. Shelves must be supported
by stanchions, not by the fish and ice below.

20
Boxing:Fish boxes come in a variety of sizes and materials. Ideally, a box should:

i. Be strong and robust.

ii. Be able to be stacked so that the weight of the top boxes are taken by the boxes
below, notby the fish in the box below.

iii. Be able to nest to save on stowage space when empty.

iv. Be easily cleaned and, if necessary, sterilized.

v. Allow ice melt-water to flow away outside the box below and not through it on to the
fish in the lower box.

vi. Have good thermal insulation.

21
Fish boxes should be used as follows
A layer of ice 5 cm thick should be placed in the bottom of the box, followed by a
layer of fish. A thin layer of ice follows, interlacing fish and ice, until the box is almost full. The
ice should be placed around the sides of the box as well as amongst the fish and the top layer of
fish should be covered with at least 5 cm of ice. The box must not be over filled. This prevents
crushing the fish when the boxes are stacked into the fish hold. When boxing fish, the fish hold
does not require any compartments or Stanchions.

Refrigerated fish rooms

Some fish rooms not only have insulation but also have refrigeration facilities. Earlier
this refrigeration is usually in the form of cooled grids and pipes on the roof of the fish room.
Now-a-days a unit cooler with fan is placed in the chill room. It is important to get the
maximum cooling effect from the ice that is allowed to melt. For this reason the temperature in
the fish room should not fall below 0oC. In order to prevent melting of ice, it is common practice
for refrigeration to be used only before fishing starts when the vessel is carrying ice alone. Under
these circumstances, the temperature can be below 0oC.

22
Insulated boxes
In small boat fisheries, insulated boxes are used to carry ice to sea, and for storing ice
and fish, when fishing occupies only a short period. The size of the boxes depends on the size of
the boat and the amount of fish normally caught in a day’s fishing. The ambient temperature will
govern the amount of insulation required in the box, though about 10-15cm of expanded
polystyrene is common. Insulated boxes for small fishing boats can often be made locally at low
cost.

Chapter 2: Calcultation of the ice requirement for cooling fish

2.2.1.Calculation of the ice requirement of cooling fish

The mass of ice needed to cool fish from the initial temperature to the final holding
temperature can be calculated from an expression, which equates the heat taken up by the ice
with the heat lost by the fish.

Heat taken up by Ice = Heat lost by fish

(Mi) (Li) = (Mf) (Cpf) (ti-tf) (1)

Where

Mi = mass of ice which melts (kg)

Li = latent heat of fusion of ice (80 kcal/kg)

Mf = mass of fish (kg)

Cpf = specific heat of fish (kcal/kgoC)

ti = initial temperature of fish (oC)

tf = final temperature of fish (0oC)

From equation (1) the ice requirement will therefore be:

(Mf) (Cpf) (ti-tf)

Mi = ----------------------- (2)

(Li)

The specific heat of lean fish is 0.8 kcal/kg oC and this value should be used if there is a
species mix or if there is a possibility that all the fish are of a lean species.

23
 The specific heat value, however, may be refined to take account of variations in the oil
content of the fish and this refined value may be used if the fish composition is reasonably
consistent.

Cpf = 0.5 Xl + 0.3 Xs + 1.0 Xw (3)

Where Cpf = specific heat of fish (kcal/kg)

Xl = mass fraction of lipids (oil)

Xs = mass fraction of solids

Xw = mass fraction of water

To illustrate the effect of oil content on the quantity of ice required for chilling the
following comparison is made between lean and fatty fish.

Example (1) – 100 kg lean fish with 1% lipids, 19% solids and 80% water at an initial
temperature of 20oC:

Cpf = (0.5 x 0.01) + (0.3 x 0.19) + (1.0 x 0.8) = 0.862 kcal/kg oC

100 x 0.862 x (20-0)

Mi = ----------------------------------- = 21.55 kg of ice

80

Example (2) – 100 kg of fatty fish with 21% lipids, 19% solids and 60% water at an initial
temperature of 20oC.

Cpf = (0.5 x 0.21) + (0.3 x 0.19) + (1.0 x 0.6) = 0.762 kcal/kgoC

100 x 0.762 x (20-0)

Mi = ----------------------------------- = 19.05 kg of ice

80

Since the calculation for fatty fish shows only a small reduction in ice requirement and,
with most species the oil content is variable, it is advisable to treat all fish as lean fish.

2.2.1.1. Heat Requirements

The heat required to change from a solid to a liquid is known as the latent heat; 1 kg of
ice requires 80 kilocalories (kcal) of heat to melt it. This figure of 80 kcal/kg is known as the

24
latent heat of fusion. It is this property of requiring a large amount of heat to melt ice that
makes it such a good cooling agent. One kilocaloryis the amount of heat required to raise the
temperature of 1kg water by 1oC.

More heat is required to warm water than almost any other substance. This capacity of
substances to hold heat, when compared to water, is known as the specific heat. Specific
heat of water is 1, for other substances it is less than 1.

Ice About 0.5

Wet fish About 0.96 (usually

taken as 1)

Frozen fish About 0.4

Air About 0.25

Most metals About 0.1

Specific heat can be used to discover how much heat has to be removed to cool a substance, i.e.
Heat to be removed = weight of substance x temperature change x the specific heat,
To cool 60 kg ice from – 5 to 10oC requires the removal of:

60 x [ - 5 – (-10)]oC x 0.5 (sp. Heat of ice) = 150 kcal

We can now calculate how much ice is needed theoretically to cool a given weight of fish:

If we want to cool 10 kg fish from 25 to 0oC, we would need to remove

10 x 25 x 1 = 250 kcal

But when ice melts it adsorbs 80kcal/kg.

250

Thus the weight of ice required = -------- = 3.12 kg.

80

This is strictly a theoretical calculation and does not take the following factors
consideration

25
 1. Ice is also melted by the surrounding air; thus a lot of ice is lost, particularly at high
ambient temperatures, unless the fish and ice are protected from the ambient heat,
preferably with insulating materials.
2. How the fish are packed in ice.
3. The length of time that the fish needs to be kept chilled once cooled.
4. How quickly the fish are chilled.

2.2.1.2. Calculation of the ice requirement for the storage of fish

Even if you are concerned with only one batch of fish held in identical containers, there are
likely to be variations in ice melting rates, which make it difficult to calculate the ice requirement
accurately. For instance, the containers are stacked, and then ice melting may be different in
containers located at the top, bottom, sides and within the stack.
All containers should be treated equally and the assumption made that each
container is fully exposed to the surrounding air.

As a first step, heat transfer may be calculated using the following simple expression:

q = A.U. (t o – tc) kcal/day (4)

Where q = heat entering the container (kcal/day)

A = surface area of the container (m2)

U = overall heat transfer coefficient (kcal/day m2oC)

to = temperature outside the container (oC)

tc = temperature inside the container (oC)

This overall calculation of heat transfer may require to be done in parts, for instance, the
lid or base of the container is made of a different material or has a different thickness. The heat
transfer through the various areas are then added together to give the total heat transfer.

The heat entering melts the ice, therefore it follows that:

q = Li .mi (kcal/day) (5)

Where q = heat required to melt ice (kcal/day)

Li = latent heat of fusion of ice (usually taken as 80 kcal/kg)

mi = mass of ice melted (kg/day)

26
 In order to develop a mathematical expression for ice melting rate during the storage
period it is assumed that ice melting inside the containers is only due to heat transferred from the
surrounding air. In this steady state approach, quantities (4) and melting (5) should be equal,
therefore it follows that:

Li . mi = A.U. (t o-tc) (6)

Thus, the ice requirement will be:

A.U. (t o-tc)

mi = -------------------- = kg/day (7)

Li

To include the element of ice melting due to radiated heat will make the calculation
extremely difficult. Therefore, if the containers cannot be protected from direct sunlight or any
other radiated heat source during the storage period, the calculated values for the ice requirement
should be upgraded.

Chapter 3: Manufacturing of different types of Ice

2.3.1. Introduction

There are many methods of producing ice. The manufacturing process of one may involve
efficiencies or advantages over another or the ice produced by one process may be better suited
to the intended application than another. Commonly used ices are block ice, flake, plate and
tube ice.

2.3.1.1. Block Ice

In the manufacture of block ice, tapered rectangular cans filled with potable water are immersed
in a tank of refrigerated brine solution .

27
28
The brine solution which is cooled to about –30oC by a refrigeration process extracts the
heat from the water and produce block ice within the can. The cans are then removed from
the tank and thawed for a short time in a tank of water to release the block from the can. The
blocks are then stored in a chill room and can be crushed on demand. The freezing period is
typically between is 16 and 24 hours although plants known as rapid blocks are available that
have freezing period of only a few hours. The quick freeze is achieved by the direct evaporation
of the refrigerant in a jacketed mould fitted with finger evaporators. The blocks are released by a
hot gas defrost and are sub-cooled below 0°C. Rapid block plant requires less floor space than
brine tank system.

Chapter 4: Super chilling (0 ºC to - 4ºC)

2.4.1. Introduction

Storage of fish at temperatures between 0oC and –4oC is called super chilling or partial
freezing. The shelf life of various fish and shellfish can be extended by storage at sub-zero
temperatures. Sub-zero storage temperatures in fishing vessels can also be obtained in
refrigerated sea water (RSW) where the freezing point of water is reduced by NaCl or
refrigerant. Compared to ice storage, the RSW systems chill fish more rapidly, reduce the
exposure to oxygen, reduce the pressures that often occur when fish are iced and also give
significant labour-saving. Super chilling extends product shelf life, but a negative effect on
freshness/prime quality has been observed for some fish species. Merritt (1965) found that cod
stored at –2oC for 10 days had an appearance and texture inferior to fish stored at 0oC in ice. The
drip of the super chilled fish was increased and at –3oC the texture of whole cod made them
unsuitable for filleting. RSW storage gives several fish species a salty taste due to the up take of
salt from sea water.

2b.4.1.1. Refrigerated Sea Water (RSW)

Seawater has a salt content of around 3 to 3.5%. At 3.5% salt, seawater has a freezing
point of about –2oC. Thus, if seawater is refrigerated, it is possible to reduce the temperature so
that a storage temperature of –1oC can be achieved. The most important advantage of RSW over
icing is the ease of handling and storage onboard, with a resultant saving of labour. In a purse -
seiner, the fish can be brailed from the net direct into the tank and brailed out of the tank at the
landing point. Pumping systems have been developed for unloading fish as large as 10kg each at
a rate of more than 25 tonnes/hour. In the most sophisticated systems, the vessel can be equipped
with a pump which pumps the fish out of the sea into holding tank and the same pumps can be
used for unloading. RSW is popular on large vessels, while CSW can be applied even on small
boats. For RSW, larger capital investment is required for the mechanical refrigeration system. In
order to avoid inefficient, too slow chilling of the catch, not more than 800kg of fish should be
packed per kilo litre of the brine. RSW may also be used to cool the catch by spraying over the
top surface of ungutted fish or shrimp in the hold. Rapid chilling and holding of the catch in
RSW is a popular practice on board large vessels which catch shrimps.

29
 Chilling of the catch in RSW before freezing on board freezer trawlers is a very
effective method of preventing rapid quality loss of the fish on subtropical and tropical
grounds. The shelf life and quality seafoods chilled or stored in CSW or RSW depends upon the
species characteristics and the condition of chilling.

Generally, it is a few days longer than that of iced fish. Some species are vulnerable
to loss of appearance due to bleaching or leaching. The uptake of salt may significantly
decrease the market value of some seafoods, like shrimps, snappers. The advantages of RSW are

1. Labour saving in storage

2. Rapid chilling, 2 to 3 times as fast as with ice
3. Elimination of crushing and ice pitting
4. Lower chill temperatures
5. Washing effect
6. Ease of unloading

2.4.1.2. Chilled sea water (CSW)

It is also possible to cool seawater by mixing ice with it. These systems are usually
referred to as chilled seawater (CSW) and can be extremely simple. A mixture of seawater and
ice, usually 1:1 to 1:2 on a volume basis at the start, forms an ice slurry that has a
temperature of about –1.5oC. In such ice slurry or chilled seawater (CSW), the heat transfer
between the fish and the cold medium occurs by convection. Thus, the rate of chilling is higher
than in ice. Ice and salt should be added to the slurry during chilling to compensate for the loss of
ice due to melting and to maintain the salt concentration at about 3%. Agitation in tank prevents
accumulation of the ice at the surface and formation of a large temperature difference between
the upper and bottom part of the container. Furthermore, forced convection increases the rate of
heat transfer.

The rate of chilling of fish in CSW is especially high in the range from ambient
temperature to about 5oC, while below 5oC, it is not significantly higher than that in crushed ice.
Thus, CSW is often used only for rapid initial chilling of highly valued fish onboard. Another
use of CSW is for chilling and holding the catch onboard in insulated containers. The shelf life of
ungutted fish kept in CSW containers is a few days longer than that of fish carefully chilled with
flake ice. Furthermore, no textural damage occurs except for some loss of scales. In the case of
small fatty fish, the CSW treatment offers some protection against rancidity. Compressed air or
nitrogen is injected into the bottom of the tank to ensure rapid and even cooling, the gas flow rate
should be 2-4kg/h at a pressure of about 35kN/m2. The contents should be agitated at least for 6
hours.

The recommended ratio of fish to seawater is between 3:1 to 4:1. In RSW systems, it
is usually necessary to provide pumped circulation of the water to ensure even mixing. This is
sometimes done with CSW systems and the motion of the vessel providing adequate mixing. A

30
rough calculation of the amount of ice needed to provide adequate cooling in a CSW system can
be made quite easily in the following manner:

Suppose 8 tonnes of fish (8,000 kg) are to be cooled from 24 to –1oC and 4:1 ratio of fish
to seawater is used, the mixture of seawater and fish would weigh 10 tonnes (10,000 kg) and the
amount of heat to be removed would be:

10,000 x 25 x 1 (taking 1 to be the specific heat of both fish and water) = 2,50,000
kcal

But, when ice melts, it absorbs 80 kcal, thus the weight of ice required:

2,50,000
---------------- = 3,125 kg
80
Thus, just over 3 tonnes of ice would be required to provide the initial cooling. This
would only reduce the temperature of the fish and would not be sufficient to maintain the lower
temperature.
Additional ice would be required to provide against heat leak. The amount of additional ice
needed would depend on the efficiency of the insulation of the tank or fish room, the
outside ambient temperature and the duration of the trip.
As a guide it may be noted that if the same 10 tonnes mixture of fish and seawater is to be
cooled from 24 to 1oC, it would impose a load of 1,000 mega joules on the refrigeration system.
If this amount of heat is to be removed in about 6 hours, a plant will have to be installed with a
power requirement of about 12 kw for the compressors.

2.4.1.3. Slurry Ice in Fish Preservation

The introduction of Deep Chill ice for direct contact cooling of fish made significant
improvements in cooling and handling processes over other forms of ice. These systems have
daily ice capacities ranging from 7 to 200 mt.
Deep Chill ice

The Deep Chill ice consists of small crystals, either in dry form or pumpable slurry, and has
the following beneficial effects:

● provides the best chilling of fish compared to other types of ice while avoiding
freezing
● the cooling process is maintained at a constant level as the ice melts
● prevents fish from bruising, because of the soft and flexible nature of ice
● easy to mix, handle and transport due to pumpability of ice slurry
● possibility of brine drainage to provide almost salt free ice for extended storage of fish

2.4.1.4. Cooling and Storage Tests

31
While studying the chilling effect of Deep Chill ice slurry with flake ice, it was found that, ice
slurry provides more efficient chilling than flake ice. This is because ice slurry effectively
eliminates hot spots in the fish container and provides maximum contact of ice surrounding
the fish.

1. It significantly retards bacterial growth and reduces fish tissue degradation.

2. The temperature drop in the fish body is extremely significant in the early stage of the
cooling process.
3. The final equilibrium temperature of the fish is approximately 10C lower, using the same
amount of ice slurry compared to flake ice.
4. The difference in cooling level occurs due to the larger enthalpy change of the ice slurry.
No uptake of salt was detected while using slurry with 0.8% salinity.

The quality of the product, such as the texture, colour and bacteria retardation is greatly
improved. Automated fish handling with ice slurry improves the productivity of the plant, and
the plant is able to deal with higher load without incurring too many shifts per day.

Chapter 5: Application of ozone / chlorine in seafood processing

2.5.1. Application of ozone in seafood processing

Pathogenic bacteria and viruses present in fish are highly sensitive to residual ozone in
water. 99.9% or higher inactivation of Aeromonas spp., Vibrio anguillarum and Yersinia ruckeri
is obtained at residual ozone concentrations of 0.15 – 0.20 mg / l within 60 seconds in natural
lake, brackish and sea water. In fish farm effluent, a residual concentration of 0.3-0.4 mg/l is
required for Aeromonas inactivation. Among fish pathogenic viruses, high sensitivity toward
ozone has generally been reported. This also applies to viruses with high UV resistance, i.e.
IPNV and WSBV as reported by many researchers.

Because of its instability, ozone must be generated and used on-site. The most efficient
method is by the electric discharge technique, which involves the passage of oxygen gas, or air,
across the gap of narrowly spaced electrodes under high voltage. Due to the energy costs of
producing ozone, it is important to optimize the transfer efficiency from gas to liquid phase.

Ozone in seafood processing and fisheries

The method of use of ozone started in late 1996 onboard purse - seiners with an idea to decrease
bacteriological problems in the circulation of RSW system.

Today, ozone is used in the seafood processing and fishery industry in areas such as:

● Purse-seiners – seawater is ozonised and circulated through RSW tanks and circulation
systems so that it comes in contact with every surface and carries out clearing.

32
 ● Well boats – when transporting live fish you have to obtain a high level of hygiene in
order to prevent mortality by pathogens. By using Ozonised Sea water to reduce
contaminants from tanks and circulation systems, and disinfects them in accordance with
applicable requirements.
● Processing plants – ozonised sea/fresh water is used for cleaning and disinfection of
process area, equipment and pipelines.
● Inlet water for fish hatcheries
● Waste water treatment – effluent from aquaculture, fish processing and fish
slaughtering. The waste water treatment systems must be tailor-made according to
regulations and are based on a combination of methods such as filtration, flotation and
separation, and ozonisation.
● Odour-ozone has been successfully used in several applications to treat odour from fish
or waste storage.

2.5.2. Use of Chlorine in Fish Processing

Chlorine is used in the fish processing sector as a water disinfectant and is probably the
most widespread disinfectant in use. Its uses include washing fishery products, addition to water
for making ice for chilling fish, and in water for thawing frozen products. It is also used in water
to cool canned fish after retorting to prevent "leaker” spoilage. Chlorine is commercially
available in several forms, the most common being a granular or powdered form as calcium
hypochlorite or in liquid form as sodium hypochlorite (NaOCl) or bleach. In any of these
forms it acts as a powerful oxidizing agent and reacts with a wide variety of compounds.
Chlorine is a gas under normal pressure and temperature. It can be compressed into a liquid and
distributed in cylinders and fed automatically into water supplies of fish processing plants or
on-board fishing vessels.

The use of chlorine dioxide is less common in fish processing. Some of the reported
advantages of chlorine dioxide over aqueous chlorine as a disinfection agent are that, it is seven
times more potent than aqueous chlorine in killing bacteria; the bactericidal activity of
chlorine dioxide is not affected by alkaline conditions and/or the presence of high levels of
organic matter. For these reasons, it is under investigation by many authorities for use in the fish
processing sector.

2.5.2.1.Concentration of chlorine used in Fish processing

It has been established in the fish processing industry that the injection of chlorine into a
supply cold water used for general wash-up helps to control microbial contamination. Chlorine
dosage should be around 10 ppm during the normal use and 100 ppm of residual concentration
during the clean – up.

33
 Codex recognize that up to 10 mg/l active chlorine in fish processing water and ice
that contacts seafood to be generally recognized as safe (GRAS).

An on-line hypochlorite injection system recommended for seawater treatment is a

dosage of 20ppm. This system, together with improved handling and storage of cooked shrimp,
has been found to double the chilled storage life while improving both the eating and
microbiological quality of products. Concentrations of 20-30 mg/L in running water have been
used to reduce the number of Listeria monocytogenes, while thawing frozen salmon. The
removal of this pathogen from the surface of fish destined for cold smoking is a major health
gain for the industry as the temperature of the cold smoking process are insufficient to kill L
monocytogenes. Chlorine at levels of 20-25 mg/L were shown to be effective in killing strains of
both Escherichia coli and L monocytogenes in a fish model system under laboratory conditions.

2.5.2.2. Marginal Chlorination

There are two important chlorination methods: marginal chlorination and

super-chlorination with or without dechlorination. The latter includes break-point chlorination
and in-plant chlorination.

Marginal chlorination is the addition of just sufficient chlorine to water to produce a

residual level. The residual chlorine may be either free or combined depending on the amount of
ammonia present in the water. This level of chlorination usually destroys pathogens.

2.5.2.3. Super Chlorination

The chlorine dosage applied is much greater than that of marginal chlorination. The
chlorine dosage depends on the character of the water and the purpose for which the water is
used. A considerable amount of chlorine may be present after a given contact period. Therefore,
for certain purposes, the water is dechlorinated. Super chlorination includes break-point
chlorination and in-plant chlorination.

2.5.2.3.1. Break-point chlorination

When small amounts of chlorine are added to water, first, chlorine reacts with certain
impurities present in the water. These impurities are substances responsible for chlorine demand.
Chlorine reacts with nitrogenous compounds to form chloramines or other chloronitrogen
compounds. On further addition of chlorine, a free residual chlorine appears. On still further
addition, the free residual chlorine gradually increases until it reaches a particular concentration.
The concentration of free chlorine is determined by the physical and chemical nature of the
water. When the level of free chlorine is increased beyond this concentration, oxidation reaction
occurs between free chlorine and chloronitrogen compounds.

The level of the free residual chlorine is decreased during the oxidation of chloronitrogen
compounds. When the oxidation is completed, the addition of chlorine to water results in
34
corresponding increase in chlorine concentration. The point after the first rise in concentration at
which the free residual reaches its lowest level is known as the breakpoint.

The advantages of break point chlorination are the following:

● Complete oxidation of ammonia and other compounds in the water

● Correction of tastes and odours of biological origin
● Correction of tastes and odours due to phenol and other substances
● Reduction of colour due to organic matter
● Improvement of bacterial quality of water

2.5.2.3.2. In-plant Chlorination

In-plant chlorination has been defined as break-point chlorination of all water as it enters the
plant to a degree where a good persisting residual chlorine occurs. In-plant chlorination of
processing water is employed in many food industries.

Chlorine gas is generally considered the best source for in-plant chlorination where large
volumes of water are to be chlorinated. But for chlorination by chlorine gas, a chlorination
equipment is needed. Hypochlorites are a good source when only small amounts of chlorine are
required. Since seafood processing industry requires only small amounts of chlorine,
hypochlorites can be used by the industry.

35
Unit 3: Freezing preservation of fish

● Chapter 1: Principle of freezing

1. Heat Units

3.1.1. Specific heat

Specific heat is the quantity of heat that must be supplied or removed from a gram of
substance to bring about change in temp of 1ºC. It is expressed in terms of kilojoules (KJ) or kcal
in applied science.

Heat capacity/specific heat of fish depends on its chemical composition; that is the
content of water, fat and minerals. It is the total heat capacity of individual components which is
given by the following formula.

Cf = Cw X W+L+Cm X m

Where,

Cf= heat capacity of fish (kcal/kg)

Cw = specific heat of water (kcal/kg)
CL = specific heat of fat/lipid (kg)
Cm = specific heat of minerals (kg)

W, L and M are contents of water, lipid and mineral in Kgs in a kg of fish.

Example : Specific heat of fish also depends on temperature. There is no applicable

change in specific heat. When fish is cooled from 30ºC to 0ºC (prior to freezing)and in this range
of temperature, specific heat of major components of fish are given as follows.

Water - 4.18 KJ/kgºC or 1 kcal/kg ºC

Fat - 2.075 KJ/kgºC or 0.5 kcal/kg ºC
Dry matter other than fat - 1.505 KJ/ kg ºC or 0.36 Kcal/ kg ºC (i.e.,
protein + minerals)

Based on these values specific heat of various fishes were calculated and found to be
between 0.74 to 0.91 Kcal / kg ºC between 30 to 0ºC. As fish gets cooled further its specific heat
goes on decreasing. Chilled fish has sp. heat of 0.845 kcal/ kg ºC which gets reduced to 0.440
kcal/ kg ºC, when the same fish is frozen. This indicates less quantity of heat is required to
change the temperature by 1ºC in frozen fish when compared to unfrozen fish. This difference is
large especially when all the water in the fish is frozen (i.e., to below -8ºC).

3.1.2. Thermal conductivity

36
 In an unevenly heat body, where there is always a difference in temperature, there is
always a flow of heat from warmer to colder layers. This type of heat exchange depends on
direct heat transfer between adjoining particles and this continues until the temperature of body
is uniform. The rate of heat transfer depends on the co-efficient of thermal conductivity of
the substance which is defined as thermal conductivity of medium in which a unit of heat
(calorie) flows through a unit surface area of unit thickness at a temperature gradient of 1
unit. Consequently expresses the amount of heat (calories) passing through a unit area of flat
surface having unit thickness at a temp difference of 1 unit in unit time. The units of thermal
conductivity are erg/cm. sec. ºC or cal/cm sec, ºC or kcal/m. Lr. ºC.

When fish is heated or cooled, the rate of conduction of heat within the fish or rate of
removal of heat depends on the co-efficient of thermal conductivity. It depends on chemical
composition of fish, physical structure, specific gravity, specific heat and temperature of fish.
The thermal conductivity in fish is important it determines the chilling, freezing and thawing
process in it. Effect of temperature on the co-efficient of their mass conductivity can be
illustrated by the following table.

Temp ºC Co-efficient of thermal

conductivity

+30°C 0.4 kcal/m.hr. ºC

0ºC 0.401-405 kcal/m.hr. ºC
-1 ºC 0.522 kcal/m.hr. ºC
-2 ºC 0.638 kcal/m.hr. ºC
-4.5 ºC 0.94 kcal/m.hr. ºC
-30 ºC 1.5 kcal/m.hr. ºC

Co-efficient of thermal conductivity in fish can be theoretically calculated if its water and dry
matter, content is known .lf=lo x XW +ln X N

lf = coefficient of thermal conductivity of fish

lo = coefficient of thermal conductivity of water in fish

ln = coefficient of thermal conductivity of dry matter in fish ( 0.22 kcal/m.hr.ºC)

W and N are respective water and dry matter %age in fish.

Co-efficient of thermal conductivity of ice is 2.07 kcal/m.hr.ºC which is almost 4 times that of
water (0.52 kcal/m.hr. ºC). Hence heat transfer in frozen fish is faster than unfrozen chilled fish.

37
 3.1.3 Thermal diffusivity

Thermal conductivity values alone will not provide accurate indication of rate of
temperature change in any given substance as these values apply solely to the rate of heat
(calories) conducted across at a constant temperature difference. Therefore, heat capacity of a
substance must be considered, as this will influence change in temperature per unit change in
heat energy. As thermal conductivity values expressed as volume basis and heat capacity on mass
basis, it is necessary to introduce a third term that is density (Mass/volume) to reconcile the two
terms. By combining all the 3 terms, we get a new term known as thermal diffusivity.
The co-efficient of thermal diffusivity “” characterizes the rate of temperature
equalization in an unevenly heated material. The C.T.D may be defined as increase in temp in
unit time when temperature gradient per unit length is equal to 1 unit (ºC)

Thermal diffusivity is related to thermal conductivity, heat capacity and specific gravity
of a material by the following expression.
= λ
----- m2/hr
cν

Where λ = coefficient of thermal conductivity k cal/m.hr.ºC

C = Specific heat in kcal/kgºC

ν = density kg/m3
Thermal diffusivity of any substance is directly proportional to its thermal conductivity
and inversely proportional to sp. heat and density of the substance. It increases as thermal
conductivity increases and density and specific. heat decreases. Also thermal diffusivity of a
substance indicates the rate at which the substance will undergo a temperature change.

Thermal diffusivity “” of a substance characterizes the speed with which the
temperature changes upon cooling or heating. The higher the coefficient, the quicker is the
change in internal temperature of the substance.

In unfrozen fish it remains almost a constant during the process of chilling (in 30ºC to
º
O C). However it changes considerably during the process of freezing and the values are very
high in completely frozen fish. This is important in frozen storage of fish because any change in
the cold storage temperature brings about marked change in temp of frozen fish. Co-efficient of
thermal diffusivity in unfrozen state is in the range of 0.000375 to 0.000475 m2/hr. The
values for fish at OoC to -3.8OC is not considered as accurate due to drastic changes in the
physical properties of fish meat in this temperature range and values are minimum in this
temperature range. In frozen fish values range from 3.75X10-4 to 15X10-4 .

3.1.4 Latent heat

It is the quantity of heat required to change the state or condition or phase under which a
substance exists without change in temp. Eg. a definite quantity of heat must be transferred to ice
38
at OOC to change it to water at the same temperature. This definite quantity of heat is known as
latent heat of fusion of water/ice.

Latent Heat of fusion of ice/water = 79.7 cal/g or 80 cal/g

It is also expressed in terms of BTU/lb
1 BTU/lb = 0.5556 kcal/kg or/k cal/kg = 1.80 BTU/lb
Latent heat of fusion of any food can be calculated if its water content is known.
Latent heat = 80X% water in food
--------------------------
100
Latent heat of fusion of some food stuffs.

Food Latent Heat of fusion

Product

BTU/lb kcal/kg

Eggs 100 55.56

Fish 101 56
Green peas 106 58.88
Milk 124 68.85
Oyster meat 125 69.44
Pork 86 47.77
Water 144 80

3.1.5 Principles of freezing

It is a common fact that the foods with high moisture content rapidly deteriorate in
quality. During freezing, the water in the food is separated out from other food components, and
is frozen. Thus, a food is protected, preserved from deteriorating influences such as temperature
and water. The lower temperature slows down the rate of chemical reaction and water is also
removed from the sphere of activity. The water is reabsorbed by the food components upon
thawing and food is restored to its original quality. However, when a food is frozen, ie. When the
water undergoes transition from liquid to solid state and also in subsequent thawing ie., when
there is transition from solid to liquid state, there is a damaging effect on food. The texture and
the structure of the food may be affected to such an extent that it may become irreversible and
upon thawing the food will not be restored to its original condition.

Fresh fish flesh normally contains 60-80% water and the freezing process converts most
of this water into ice. At normal atmospheric pressure, pure water will change from liquid to
solid (ice) at 0oC, i.e., it will freeze. The water in fish flesh contains salts and chemicals, which
have the effect of lowering the temperature at which the water begins to freeze. The water in fish
flesh begins to freeze at about –1oC and as the temperature drops below –1oC, more water is
frozen out and the concentration of salts in the remaining water rises, so that its freezing point is
39
lowered further. At –5oC, it would appear that all the water is frozen, but over 20% of the water
in fish muscle is still unfrozen. Even at –30oC, approximately 10% of the water remains
unfrozen.The above defined heat units are applied here.

In order to change the physical state of a substance from a liquid to a solid, energy or
latent heat has to be removed from the substance. In order to lower the temperature of 1g of
water by 1oC, at temperatures above 0oC, 1 calorie of heat must be removed this is known as
the “specific heat”. However, to change water at 0oC to ice at 0oC, 80 calories must be
removed for each g of water. In other words, the specific heat of liquid water is 1 and the latent
heat of changing liquid water to ice is 80. The specific heat of ice at temperatures below 0oC is
0.5, which means that to lower the temperature of ice by 1oC, 0.5 calories of heat needs to be
removed. For all practical purposes, it is assumed that fish have the same values for specific heat
and latent heat as water. All this means that, if heat is removed from fish at a constant rate, there
will be a period during the fish freezing, where the temperature of fish will not drop. This period
lasts until approximately 75% of the water is frozen, when the temperature begins to drop again.
There are three stages to freezing fish fig.1). During stage 1, the temperature falls fairly rapidly
to just below 0oC (and it is called as initial falling rate period), during stage 2, the temperature
remains fairly constant at about –1oC as the bulk of the water (three quarters of the water) in the
fish freezes (this stage is known as the ‘thermal arrest’ period) and during stage 3 (final falling
rate period), the temperature again drops and most of the remaining water becomes frozen. The
temperature at –1.1oC, freezing begins, and –5oC, about 80% of water is frozen, this range is
termed at ‘Critical range’. The ideal thermal arrest is only 30min. Freezing is complete only
when the equilibrium temperature reaches –18oC(ie. the process should not be regarded as
complete unless and until the product temperature has reached –18oC at the thermal centre after
thermal stabilization). It also shows that even at temperatures as low at –30oC, a portion of water
in the fish muscle still remains in the unfrozen state.

Under ideal conditions, fish should be frozen to –30oC in 2 hours.

Using simple mathematics, the amount of energy required to freeze fish can be
calculated.
Suppose, if 1 kg fish at 25oC is to be frozen at –30oC. During stage 1, one calorie of
energy has to be extracted for each gram material for each drop of 1oC. In an example, the
temperature of 1000g of fish will be lowered from 25oC to –1oC, ie., by 26oC. The energy
required will be equal to 1000 x 26 x the specific heat of water (1) = 26000 calories or 26
kilocalories (kcal).
During stage 2, 80 calories of energy needs to be extracted for each gram material frozen. In the
example, 1000g of fish are to be frozen. The energy required will be equal to 1000 x the latent
heat for freezing water (80), which equals 80,000 calories or 80kcal.
During stage3, 0.5 calories of energy needs to be extracted for each g material for each
1oC drop in temperature. In the example, the temperature of 1000g of fish will be lowered from
–1 to –30oC, ie., by 29oC. The energy required will be equal to 1000 x 29 x the specific heat of
ice (0.5 = 14500 calories =14.5 kcal.

To summaries:
Stage 1, 1000 x 26 x 1 = 26 kcal

40
 Stage 2, 1000 x 80 = 80 kcal
Stage 3, 1000 x 29 x 0.5 = 14.5 kcal
Adding these three figures gives 120.5 kcal, ie., to freeze 1kg of fish from 25oC to
–30oC, 120.5 kcal of energy is required.

From the above example, it is apparent that more than 50% of the energy extraction
during freezing of fish is taking place in stage 2. The thermal arrest period, where little or no
drop in temperature is occurring, and this period is a critical one, if a good frozen product is to be
produced. Ideally, fish should pass through the thermal arrest period as quickly as possible, due
to the following reasons:

1. It is postulated that slow freezing produces large ice crystals in the cells of the fish, which
can be larger than the cells themselves and so break the cell walls. But it is not so,
because it is evident that the walls of fish muscle cells are sufficiently elastic to
accommodate the larger ice crystals without excessive damage. Also, most of the water in
fish muscle is bound to the protein in the form of a gel and little fluid would be lost.

2. As the water begins to freeze in the flesh, the concentration of salts and chemicals in the
remaining water rises. This high concentration of salts and enzymes can cause
accelerated autolysis and denaturation of protein during slow freezing, This result in an
inferior quality product and it is mainly due to denaturation of the protein. Changes take
place in some fractions of the protein as a result of freezing and since the proteins are
altered from their “native” state they may be said to be “denatured”, hence the term
“protein denaturation”.

3. At temperature around 0oC, certain types of bacteria are still active and bacterial spoilage
can still occur.

4. Textural changes occur in fish, which have been frozen slowly, caused by the presence of
large ice crystals and denaturation of protein during the accelerated autolysis. In addition,
a phenomenon known as ‘thaw drip’ occurs, when slowly frozen fish are thawed. On
thawing, the water which was originally bound within the cells are released and
considerable loss in weight can occur.

However, from a textural point of view, it is unlikely that a highly trained taste panel
could detect the difference between fish frozen in 1 hour and those frozen in 8 hours, Freezing
times of 24 hours or more will almost certainly result in an inferior product and very long
freezing times can result in bacterial spoilage making the fish unfit for consumption.

41
Chapter 2: Physical, chemical and thermodynamic properties of refrigerants

3.2.1. Refrigerant

It is any substance that absorb heat through expansion or vaporization and loses it threw
condensation in a refrigeration system. It is otherwise called as working fluid in a
refrigeration system

Desirable properties of an ideal Refrigerant

1. Low boiling point
2. High critical temperature
3. High latent heat of vaporization
4. Low specific volume of vapour
5. Non-corrosive to metal
6. Non-flammable and non-explosive
7. Non – toxic
8. Low cost
9. Easy to liquefy at moderate pressure and temperature
10. Easy of locating leaks
11. Mixes well with oil
A refrigerant is chosen which has larger advantages and less disadvantages.

3.2.2. Thermodynamic properties

1. Boiling temperature

Should be low at atmospheric pressure. If it is high, compressor should be operated at high

vaccum, reduces capacity and increase the cost of the system.

e.g.

Refrigerant Boiling temperature (ºC)

R-12 (CCl2F2) - 29

R-717 (NH4) -33.3

R.744 (CO2) -73.6

R-764 (SO2) -10

2. Freezing temperature:

42
 Should be below the operating temperature of evaporator. Freezing temperature of most of
refrigerants are below -35ºC

Refrigerant Boiling temperature (ºC)

R-12 - 157.5

R-717 -35

R.744 - 56.7

R-764 - 75.6

3. Evaporator and condenser pressure

Both the evaporating and condensing pressure should be +ve (i.e. above atmospheric
pressure). The + ve pressure are necessary to prevent leakage of air, moisture into the
refrigeration systems. It also permits easier detection of leaks.

Critical temperature & pressure:

It is the highest temperature at which refrigerant can be condensed to liquid, regardless

of high pressure.

Refrigerant Critical temperature (ºC) Critical pressure (bar)

R-12 112 41.2

R-717 133 113.86

R.744 31 73.8
R-764 157 78.7

Coefficient of performance and power requirements

For an ideal refrigerant operating between -15ºC evaporator temperature and 300C
condenser temperature. The theoretical coefficient of performance for the reversed cycle is 5.74.

Refrigerant Coefficient of performance Power HP/TR

R-12 4.70 1.00

R-11 5.09 0.93
R-717 4.76 0.99
R.744 2.56 1.84
R-764 4.87 0.97

43
 3.1.3 Thermal diffusivity

Thermal conductivity values alone will not provide accurate indication of rate of
temperature change in any given substance as these values apply solely to the rate of heat
(calories) conducted across at a constant temperature difference. Therefore, heat capacity of a
substance must be considered, as this will influence change in temperature per unit change in
heat energy. As thermal conductivity values expressed as volume basis and heat capacity on mass
basis, it is necessary to introduce a third term that is density (Mass/volume) to reconcile the two
terms. By combining all the 3 terms, we get a new term known as thermal diffusivity.

The co-efficient of thermal diffusivity “” characterizes the rate of temperature

equalization in an unevenly heated material. The C.T.D may be defined as increase in temp in
unit time when temperature gradient per unit length is equal to 1 unit (ºC)

Thermal diffusivity is related to thermal conductivity, heat capacity and specific gravity
of a material by the following expression.
= λ
----- m2/hr
cν

Where λ = coefficient of thermal conductivity k cal/m.hr.ºC

C = Specific heat in kcal/kgºC

ν = density kg/m3

Thermal diffusivity of any substance is directly proportional to its thermal conductivity

and inversely proportional to sp. heat and density of the substance. It increases as thermal
conductivity increases and density and specific. heat decreases. Also thermal diffusivity of a
substance indicates the rate at which the substance will undergo a temperature change.

Thermal diffusivity “” of a substance characterizes the speed with which the
temperature changes upon cooling or heating. The higher the coefficient, the quicker is the
change in internal temperature of the substance.

In unfrozen fish it remains almost a constant during the process of chilling (in 30ºC to
º
O C). However it changes considerably during the process of freezing and the values are very
high in completely frozen fish. This is important in frozen storage of fish because any change in
the cold storage temperature brings about marked change in temp of frozen fish. Co-efficient of
thermal diffusivity in unfrozen state is in the range of 0.000375 to 0.000475 m2/hr. The
values for fish at OoC to -3.8OC is not considered as accurate due to drastic changes in the
physical properties of fish meat in this temperature range and values are minimum in this
temperature range. In frozen fish values range from 3.75X10-4 to 15X10-4 .

3.1.4 Latent heat

44
 It is the quantity of heat required to change the state or condition or phase under which a
substance exists without change in temp. Eg. a definite quantity of heat must be transferred to ice
at OOC to change it to water at the same temperature. This definite quantity of heat is known as
latent heat of fusion of water/ice.
Latent Heat of fusion of ice/water = 79.7 cal/g or 80 cal/g
It is also expressed in terms of BTU/lb
1 BTU/lb = 0.5556 kcal/kg or/k cal/kg = 1.80 BTU/lb
Latent heat of fusion of any food can be calculated if its water content is known.
Latent heat = 80X% water in food
--------------------------
100
Latent heat of fusion of some food stuffs.

Food Latent Heat of fusion

Product

BTU/lb kcal/kg

Eggs 100 55.56

Fish 101 56
Green peas 106 58.88
Milk 124 68.85
Oyster meat 125 69.44
Pork 86 47.77
Water 144 80

3.1.5 Principles of freezing

It is a common fact that the foods with high moisture content rapidly deteriorate in
quality. During freezing, the water in the food is separated out from other food components, and
is frozen. Thus, a food is protected, preserved from deteriorating influences such as temperature
and water. The lower temperature slows down the rate of chemical reaction and water is also
removed from the sphere of activity. The water is reabsorbed by the food components upon
thawing and food is restored to its original quality. However, when a food is frozen, ie. When the
water undergoes transition from liquid to solid state and also in subsequent thawing ie., when
there is transition from solid to liquid state, there is a damaging effect on food. The texture and
the structure of the food may be affected to such an extent that it may become irreversible and
upon thawing the food will not be restored to its original condition.

Fresh fish flesh normally contains 60-80% water and the freezing process converts
most of this water into ice. At normal atmospheric pressure, pure water will change from liquid
to solid (ice) at 0oC, i.e., it will freeze. The water in fish flesh contains salts and chemicals,
which have the effect of lowering the temperature at which the water begins to freeze. The water

45
in fish flesh begins to freeze at about –1oC and as the temperature drops below –1oC, more water
is frozen out and the concentration of salts in the remaining water rises, so that its freezing point
is lowered further. At –5oC, it would appear that all the water is frozen, but over 20% of the
water in fish muscle is still unfrozen. Even at –30oC, approximately 10% of the water
remains unfrozen.The above defined heat units are applied here.

In order to change the physical state of a substance from a liquid to a solid, energy or
latent heat has to be removed from the substance. In order to lower the temperature of 1g of
water by 1oC, at temperatures above 0oC, 1 calorie of heat must be removed this is known as
the “specific heat”. However, to change water at 0oC to ice at 0oC, 80 calories must be
removed for each g of water. In other words, the specific heat of liquid water is 1 and the latent
heat of changing liquid water to ice is 80. The specific heat of ice at temperatures below 0oC is
0.5, which means that to lower the temperature of ice by 1oC, 0.5 calories of heat needs to be
removed. For all practical purposes, it is assumed that fish have the same values for specific heat
and latent heat as water. All this means that, if heat is removed from fish at a constant rate, there
will be a period during the fish freezing, where the temperature of fish will not drop. This period
lasts until approximately 75% of the water is frozen, when the temperature begins to drop again.
There are three stages to freezing fish fig.1). During stage 1, the temperature falls fairly rapidly
to just below 0oC (and it is called as initial falling rate period), during stage 2, the temperature
remains fairly constant at about –1oC as the bulk of the water (three quarters of the water) in the
fish freezes (this stage is known as the ‘thermal arrest’ period) and during stage 3 (final falling
rate period), the temperature again drops and most of the remaining water becomes frozen. The
temperature at –1.1oC, freezing begins, and –5oC, about 80% of water is frozen, this range is
termed at ‘Critical range’. The ideal thermal arrest is only 30min. Freezing is complete only
when the equilibrium temperature reaches –18oC(ie. the process should not be regarded as
complete unless and until the product temperature has reached –18oC at the thermal centre after
thermal stabilization). It also shows that even at temperatures as low at –30oC, a portion of water
in the fish muscle still remains in the unfrozen state.
Under ideal conditions, fish should be frozen to –30oC in 2 hours.

Using simple mathematics, the amount of energy required to freeze fish can be calculated.

Suppose, if 1 kg fish at 25oC is to be frozen at –30oC. During stage 1, one calorie of

energy has to be extracted for each gram material for each drop of 1oC. In an example, the
temperature of 1000g of fish will be lowered from 25oC to –1oC, ie., by 26oC. The energy
required will be equal to 1000 x 26 x the specific heat of water (1) = 26000 calories or 26
kilocalories (kcal).
During stage 2, 80 calories of energy needs to be extracted for each gram material frozen. In the
example, 1000g of fish are to be frozen. The energy required will be equal to 1000 x the latent
heat for freezing water (80), which equals 80,000 calories or 80kcal.

During stage3, 0.5 calories of energy needs to be extracted for each g material for each
1oC drop in temperature. In the example, the temperature of 1000g of fish will be lowered from
–1 to –30oC, ie., by 29oC. The energy required will be equal to 1000 x 29 x the specific heat of
ice (0.5 = 14500 calories =14.5 kcal.

46
 To summaries:
Stage 1, 1000 x 26 x 1 = 26 kcal
Stage 2, 1000 x 80 = 80 kcal
Stage 3, 1000 x 29 x 0.5 = 14.5 kcal

Adding these three figures gives 120.5 kcal, ie., to freeze 1kg of fish from 25oC to
–30oC, 120.5 kcal of energy is required.

From the above example, it is apparent that more than 50% of the energy extraction
during freezing of fish is taking place in stage 2. The thermal arrest period, where little or no
drop in temperature is occurring, and this period is a critical one, if a good frozen product is to be
produced. Ideally, fish should pass through the thermal arrest period as quickly as possible, due
to the following reasons:

1. It is postulated that slow freezing produces large ice crystals in the cells of the fish, which
can be larger than the cells themselves and so break the cell walls. But it is not so,
because it is evident that the walls of fish muscle cells are sufficiently elastic to
accommodate the larger ice crystals without excessive damage. Also, most of the water in
fish muscle is bound to the protein in the form of a gel and little fluid would be lost.
2.
3. As the water begins to freeze in the flesh, the concentration of salts and chemicals in the
remaining water rises. This high concentration of salts and enzymes can cause
accelerated autolysis and denaturation of protein during slow freezing, This result in an
inferior quality product and it is mainly due to denaturation of the protein. Changes take
place in some fractions of the protein as a result of freezing and since the proteins are
altered from their “native” state they may be said to be “denatured”, hence the term
“protein denaturation”.

4. At temperature around 0oC, certain types of bacteria are still active and bacterial spoilage
can still occur. .

5. Textural changes occur in fish, which have been frozen slowly, caused by the presence of
large ice crystals and denaturation of protein during the accelerated autolysis. In addition,
a phenomenon known as ‘thaw drip’ occurs, when slowly frozen fish are thawed. On
thawing, the water which was originally bound within the cells are released and
considerable loss in weight can occur.

However, from a textural point of view, it is unlikely that a highly trained taste
panel could detect the difference between fish frozen in 1 hour and those frozen in 8 hours,
Freezing times of 24 hours or more will almost certainly result in an inferior product and
very long freezing times can result in bacterial spoilage making the fish unfit for
consumption.

Chapter 2: Physical, chemical and thermodynamic properties of refrigerants

47
 3.2.1. Refrigerant

It is any substance that absorb heat through expansion or vaporization and loses it threw
condensation in a refrigeration system. It is otherwise called as working fluid in a
refrigeration system

Desirable properties of an ideal Refrigerant

1. Low boiling point
2. High critical temperature
3. High latent heat of vaporization
4. Low specific volume of vapour
5. Non-corrosive to metal
6. Non-flammable and non-explosive
7. Non – toxic
8. Low cost
9. Easy to liquefy at moderate pressure and temperature
10. Easy of locating leaks
11. Mixes well with oil
A refrigerant is chosen which has larger advantages and less disadvantages.

3.2.2. Thermodynamic properties

1. Boiling temperature

Should be low at atmospheric pressure. If it is high, compressor should be operated at high

vaccum, reduces capacity and increase the cost of the system.

e.g.

Refrigerant Boiling temperature (ºC)

R-12 (CCl2F2) - 29

R-717 (NH4) -33.3

R.744 (CO2) -73.6

R-764 (SO2) -10

2. Freezing temperature:

48
 Should be below the operating temperature of evaporator. Freezing temperature of most of
refrigerants are below -35ºC

Refrigerant Boiling temperature (ºC)

R-12 - 157.5

R-717 -35

R.744 - 56.7

R-764 - 75.6

3. Evaporator and condenser pressure

Both the evaporating and condensing pressure should be +ve (i.e. above atmospheric pressure).
The + ve pressure are necessary to prevent leakage of air, moisture into the refrigeration systems.
It also permits easier detection of leaks.

Critical temperature & pressure:

It is the highest temperature at which refrigerant can be condensed to liquid, regardless of high
pressure.

Refrigerant Critical temperature (ºC) Critical pressure (bar)

R-12 112 41.2

R-717 133 113.86

R.744 31 73.8
R-764 157 78.7

Coefficient of performance and power requirements

For an ideal refrigerant operating between -15ºC evaporator temperature and 300C condenser
temperature. The theoretical coefficient of performance for the reversed cycle is 5.74.

Refrigerant Coefficient of performance Power HP/TR

R-12 4.70 1.00

R-11 5.09 0.93
R-717 4.76 0.99
R.744 2.56 1.84
R-764 4.87 0.97

49
 3.2.3. Latent heat of vaporization

A refrigerant should have a high latent heat of vapour at the evaporator temperature. It
results in high refrigerating effect per kg of refrigerant and which reduces the mass of refrigerant
to be circulated per ton of refrigerants.

Refrigerant Latent heat of vapour at 15ºC

(Kcal/kg)

R-12 (CCl2F2) 37.97

R-717 (NH4) 314.42
R.744 (CO2) 65.44
R-764 (SO2) 94.27

Specific volume

It indicates the theorentional displacement of the compressor (i.e volume of suction vapour to
compressor).

3.2.4 Physical properties of refrigerants

Stability and inertness

An ideal refrigerant should not decompose at any temperature normally encountered in the
refrigerating system. This is due to the reaction with metal. To avoid this, a refrigerant should be
inert with respect to all material used in refrigerated system.

Corrosive property

Freon groups are non-corrosive with all metals. NH4 is used only with steel and Sn. S02 is
non-corrosive to all metals in the absence of water because it reacts with water and forms
sulphuric acid.

Viscosity:

The refrigerant in the liquid and vapour states should have low viscosity because the heat
transfer through condenser and evaporator is improved at low viscosities.

Thermal conductivity

The refrigerant in the liquid and vapour states should have high thermal conductivity. This is
required in finding the heat transfer coefficient in evaporator and condensers.

Leakage tendency:

50
It should be low and the leakage of refrigerant should be easily detected by its pungent odour and
also by using burning sulphur candle which forms white fumes of ammonium sulphate in the
presence of ammonia.

Cost

It is not so important for small refrigerant units but it is very much important in high capacity
refrigerant system like industry and commercial. Ammonia is the cheapest one. So it is widely
used in large industrial plant such as cold storage and ice plants. R-22 is costlier than R-12. The
cost of loss due to leakage is also important.

3.2.5. Refrigerant

It is a substance in the vapour state which is used as a working fluid in refrigerators. In a

refrigerating machine, heat is transferred from cold body to hot body through refrigerants. It is
also a cooling agent of refrigerating system. The choice of refrigerant depends on ability to
remove heat, toxicity, density, availability, cost etc.

Classification

1. Primary refrigerants
2. Secondary refrigerants

Primary refrigerants

These are the refrigerants which cool the substance or space directly by absorbing latent heat. It
is also known as direct expansion system Eg. Ammonia, Freon, SO2, CO2 etc.

Ammonia (NH3)

o Used for commercial purposes. Mainly in cold stored and ice plants.
o The boiling temperature of NH3 is -33oC
o It is colorless gas with a sharp pungent smell
o Has good thermodynamic properties
o It is neutral to all metals, highly soluble in oil.
o It’s solubility increases with increasing pressure and decreasing temperature
o Used free small and medium refrigerating capacities
o Volatile and non toxic but in higher concentration suffocation occur due to lack of
O2

SULPHUR DIOXIDE (SO2)

51
 o Previously used in household refrigerators
o Toxic, non-explosive and non-flammable, non-corrosive
o Irritant to human body
o Non mixable with oil
o Has pungent odour and low latent heat value

CARBON DI OXIDE (CO2)

o Used as dry ice (solid from of CO2)

o Not used as refrigerant
o Colourless, odourless, non-toxic, non explosive, non-corrosive, non-flammable
o Operating pressure is very high
o Not mixable with oil
o As it is colourless and odourless gas, leaks cannot be detected easily
o Safest of all
o Its thermodynamic properties are not desirable for refrigeration except in
cryogenic applications
o Two forms of CO2 used to freeze foods are spraying liquid CO2& CO2 snow.

3.2.6 Freon group

o They are fluorocarbons of methane and ethane series.

o They contain 1 or more of these halogens (chlorine, bromine, fluorine)
o Non toxic, non-flammable, non-explosive, non- corrosive, non-irritant to human
body and eyes.
o Odourless, colourless
o Will not react with food product stored in the refrigerated space.
o Will not react with lubricating oil.
o Has excellent thermodynamic properties
o Only disadvantage is ozone layer is damaged.

FREON 11 (Trichloro monofluro methane – CCl3F)

o Has the boiling point of 74.8ºF.

o Used in air conditioning plants in a larger level factories, theatres

FREON 12 (Dichloro-difluro methane –CCl2F2)

o Mostly used in domestic and commercial refrigeraters (in ice cream cabinets,
display cabinets, deep freezer)
o It is very widely used, colourless gas with mild odour
o Heavier than air
o Does not dissolve in water, moisture content should not exceed 0.0025% by
weight
o It does not react with metals like ferrous, aluminium, phosphor bronze

52
 o It attacks copper, copper alloys, zinc and bronze and dissolves in water
o It does not react with lubricating oils in the absence of moisture, but oxidizes
them in the presence of water vapour

FREON 13 (Mono chloro-trifluromethane) (CCl F3)

o Widely used
o Somewhat inferior in thermodynamic properties
o FREON 21 – (dichloro - monofluoroethane) (CHCl2F)
o FREON 22 – (monochloro – diflueroethane) (CHClF2)
o There are used in small refrigeration plants
o Used to obtain moderately low temperature (-50ºC)

3.2.7 Secondary refrigerant

● Substances that take away heat from the medium to be cooled and give it to the boiling
refrigerant are called secondary refrigerants. They do not change their physical condition
● Cheap and harmless, chemically neutral to metals and packing material
● Should have a low freezing point and large heat capacity
● The refrigerants are brine which is used as intermediate fluid between evaporator and the
substance or space to be cooled. They cool the substance and the space by absorbing their
sensible heat, which is also being called having indirect expansion system.
● Eg. Brine solution made of calcium chloride or sodium chloride
● Choice of brine depends on temperature to which a material is to be cooled and industrial
process in which it is to be used
● For lower temperature (-18 to -23oC), CaCl2 is used, it is very costly
● NaCl is used when it is desired to have direct contact between brine and product.
● Freezing point of brine depends on its concentration
● Cheapest secondary refrigerants are water and air but their application is limited. Since,
water has a high freezing point (0oC) and air has a low heat capacity.

3.2.8. Physical characteristics of refrigerants

o Boiling and condensing temperature and pressures

o Freezing temperature
o Critical temperature
o Discharge temperature
o Latent heat of vapourisation
o Specific heat
o Density
o Viscosity

53
Boiling and condensing temperature and pressures

o The evaporator and condensing temperatures determine the pressures

o The maximum condensing temperature is largely affected by climatic condition
o It is desirable to select a refrigerant whose saturation pressure (at minimum
operating temperature) is a few pounds above atmospheric pressure.

Freezing temperature

o Should have low freezing temperature to avoid operational obstruction by the

refrigerant itself

Critical temperature

o Should be well above the maximum condensing temperature

Discharge temperature

o High discharge temperatures from the compressor should be avoided

o It causes some refrigerant breakdowns as well as poor lubrication effectiveness

Latent heat of vapourization

o Heat which converts the refrigerant from the liquid state to vapour
o It should have a higher value

Specific heat

o Higher specific heat is not desirable

Density

o Low vapour density refrigerants are preferred

Viscosity

o Low viscosity of the liquid refrigerant is desired to reduce the pressure drop in the
lines

3.2.9. Chemical properties of refrigerants

o Toxicity
o Flammability and explosion hazard
o Refrigerant odours

54
Toxicity

o Rated based on its effect on human beings over specified periods

o Should be non toxic and non irritation

Flammability and explosion Hazard

o Should be non-flammable and non-explosive

Refrigerant Odours

o Can be both an asset and a hazard

o Makes it easy to detect the leaks but at the same time may contaminate foodstuffs
in storage

Chemical properties

o Should be non-flammable
o Should be non-explosive
o Should be not-toxic
o Should not react with lubricating oil
o Should not react with moisture
o Should not contaminate the food materials kept inside the refrigerating system

3.2.10. Chemical properties

1. Flammability

It is an important characteristic in the selection of refrigerant. Ammonia and methyl chloride

refrigerant will burn but are explosive under unusual conditions only. CO2, N2, NO2 and
fluorocarbon are nonflammable and non-explosive.

2. Toxicity

All fluorocarbon refrigerant become toxic when mixed with certain percentage of air. Ammonia
and SO2 are highly toxic and not used in air conditioning. The use of toxic refrigerant is limited
to cold storage.

3. Miscibility

The ability of refrigerant to mix with oil is called miscibility. The degree of miscibility depends
upon the temperature of oil and pressure of refrigerating vapour. The Freon group are highly
miscible refrigerant while ammonia, CO2, SO2, and methyl chloride are relatively non-miscible.

55
The non – miscible refrigerant require larger heat transfer surface due to poor heat conduction
properties of oil. The miscible refrigerant give better lubrication.

4. Effect on perishablematerials:

The refrigerant used in cold storage and in domestic refrigerators should be such that in case of
leakage, it should have no effect on the perishable material.

The Freon group have no effect on dairy products, meats, vegetable, flowers & fruits. Methyl
chloride also have no effect on materials. But SO2, ammonia affect the material.

NH4 easily dissolve in water and becomes alkaline in nature.

Since most fruits and vegetables are acidic in nature, therefore ammonia reacts with these
products and spoils the taste.

3.2.11. Economic and thermodynamic properties

● Should have high value of latent heat of vaporization

● Should have high thermal conductivity
● Should have high electrical resistance
● Should have low specific volume
● Should have low specific heat
● Should have low freezing point (than the maximum temperature of the refrigeration
system)
● Should have low boiling temperature
● Should have low leakage tendency. Leakage should be detectable
● Above all the refrigerants should not be expensive

56
Chapter 3: Types of freezer

3.3.1. Introduction

The three basic methods of freezing fish are:

1. Blowing a continuous stream of cold air over the fish – air blast freezers
2. Direct contact between the fish and a refrigerated surface – contact or plate freezers
3. Immersion in or spraying with a refrigerated liquid – immersion or spray freezers

4. 3.3.1.1. Air blast freezers

The advantage of the blast freezer is its versatility. It can cope with a variety of irregularly
shaped products and whenever there is a wide range of shapes and sizes to be frozen, the
blast freezer is the best choice. Before going on to describe the various types of air blast
freezer, it is necessary to deal with some of the basic principles of air blast freezer design and
operation.

Designing air blast freezers

The use of air to transfer heat from the product being frozen to the refrigeration system is
probably the most common method used in commercial refrigeration. The natural convection of
the air alone would not give a good heat transfer rate; therefore, forced convection by means of
fans has to be introduced. To enable the product to be frozen in a reasonable time the air flow
rate should be fairly high. Also, in order to obtain uniform freezing rates throughout the freezer,
the air flow requires to be consistent over each fish or package.

Frost build-up on the cooler is also more prolific on the upstream coils; therefore a cooler with a
large frontal area will be able to operate longer before a defrost is necessary.

Continuous air blast freezers

The belt has to be flexible, easily cleaned, non corroding, suitable for use in direct contact with
food and should not interfere unduly with either the freezing time or adversely affect product
quality. Stainless steel mesh link belts or chain link belts are mainly used for this purpose but
they have certain disadvantages. Apart from being expensive, they affect the appearance of the
product. If fish are loaded directly on the belt, the crinkled or indented appearance of the frozen
product is not always acceptable. Open mesh belts can also give rise to difficulty when removing
the product after freezing and some weight loss may be incurred due to slight physical damage.
Skin-on fillets can usually be removed quite easily but skinless fillets and fish portions can stick
to the belt and cause unacceptable weight losses.

Continuous belt freezers can be constructed with either cross-flow or series-flow air circulation.
In the series-flow arrangement, the direction of air flow must be such that the coldest fish meet

57
the coldest air. The design of the belt entry and exit must keep the rate of air infiltration to a
minimum.

Fluidized and semi-fluidized freezers

One type of air blast freezer fluidizes the product with a strong blast of air from below. The
product then behaves like a fluid and when poured into the trough at the input, it moves along the
length of the freezer without mechanical assistance and over-flows at the output. This type of
freezer has not had a wide application for fish or fishery products. Small cooked and shelled
shrimp is one of the few fish products that have been successfully frozen by this method.

3.3.1.2. Plate freezers

Plate freezers can be arranged with the plates horizontal to form a series of shelves and,
as the arrangement suggests, they are called horizontal plate freezers (HPF). When the plates are
arranged in a vertical plane they form a series of bins and in this form they are called vertical
plate freezers (VPF). Modern plate freezers have their plates constructed from extruded sections
of aluminium alloy arranged in such a manner as to allow the refrigerant to flow through the
plate and thus provide heat transfer surfaces on both sides. Plate freezers are fitted with hydraulic
systems which move the plates together and apart.

Horizontal plate freezers

At present in India all the processing plants have HPF. Capacity of this freezer varies
between 500kg to 1ton/load. Each plates are connected with pressure resistant hose on both ends,
i.e. suction line and discharge line. A hot gas defrost arrangement is the quickest method to

58
defrost an HPF, but even with this method, it may take 30 min or more. The defrosted plates
must be completely free from frost or ice and dried before the freezer is used again.

Vertical plate freezers

The main advantage of this type of freezer is that fish can be frozen in bulk without the
requirement to package or arrange on trays. This type of freezer is therefore particularly suitable
for bulk freezing and it has also been extensively used for freezing whole fish at sea. The
maximum size of block made by this method is usually 1070 nm x 535 mm. Other dimensions
however, can be produced in which the thickness can vary from 25 to 130 mm, and it will
depend on the fish to be frozen. The maximum weight and dimensions are also limited by the
physical effort required from the operator to lift the block, and by the ease with which it can be
handled so that damage to the fish is kept to a minimum.

In most cases, fish can be loaded between the plates without wrappers and water need not
be added either to strengthen the frozen block or improve the contact with the plates. Fish such
as cod and haddock produce compact blocks with a block density of approximately 800 kg/m3. It
has certain operational disadvantages.

Defrost is essential to prevent fish sticking to the plates which are at a temperature below
0oC, and thus failing to form a compact block. Freezing times are longer due to the poor contact
being made with the plates and because of the lower block density, more storage space is
required for a given quantity of fish.

3.3.1.3. Liquid nitrogen freezer

In this freezer, the product is brought into direct contact with the refrigerant. The fish on
the stainless steel conveyor belt initially come into contact with the counter current flow of
nitrogen gas at a temperature of about –50oC. As the fish progress through the pre-cooling stage
of the freezer, the gaseous nitrogen partially freezes the fish and up to 50 percent of the product
heat is extracted. The product then passes below the liquid spray where freezing is completed by
the boiling liquid. The last stage in the freezer provides a few minutes for the fish temperature to
reach equilibrium before the fish are discharged.

The main advantage of the liquid nitrogen freezer LNF is that freezing is very quick and
the physical size of the freezer is correspondingly small. The freezer is operated without the
need for compressors, condensers or coolers; therefore maintenance requirements are
minimal and the power required to operate the freezer is very low. Liquid nitrogen must be
retained in a vacuum insulated pressure vessel with continuous venting to keep the contents cool
and the internal pressure down. It is found that 0.5 percent of the stored contents is lost each day
by this method. In addition, about 10 percent has been estimated to be lost during the transfer of
liquid from the tanker to the storage vessel. This method of freezing is more expensive than most
others. It is four times more costly than conventional air blast freezing. Although the freezer
is small and there is no refrigeration machinery requirement, storage space and access is required
for the liquid nitrogen tank. The main disadvantage of this type of freezer in most developing

59
countries like India is that delivery of nitrogen could be expensive and there may be no guarantee
of regular supplies.

3.3.1.4. Carbon dioxide freezer

This type of freezer has been known for a long time and uses liquefied carbon dioxide
which is usually a by-product of another industrial process.

The liquefied carbon dioxide is injected into the freezer has direct contact with the
product. In this respect, it is similar in operation to an LNF. With large units, it is economically
feasible to recover the carbon dioxide and about 80 percent of the refrigerant used can be
reliquefied. Carbon dioxide can be contained in insulated vessels at a moderate pressure and
losses during storage are therefore negligible. High levels of carbon dioxide in the factory air are
dangerous, therefore a freezer using this refrigerant must be vented and the gas discharged
outside the building. Again, as in the case with other types of freezer which rely on regular
supplies of refrigerant, carbon dioxide freezers would not be suitable for use in remote areas.

3.3.1.5. Immersion freezers

In this method a liquid refrigerant is used for the removal of heat from a product.
Favourable freezing rates can be achieved. Liquid can remove more heat per unit volume than
gas (eg. air) but, like gas, a stagnant boundary layer is form which slows the transfer of the heat.
Liquids used for heat transfer must therefore be circulate over the product. Difficulties due to
high viscosity often arise when a low temperature liquid is used.

60
Chapter 4: Methods of protective treatments

3.4.1. Brining

To minimize drip loss, fillets/steaks of fish are immersed in brine for a short time and this
process is known as firming of fillets/steaks. There are several advantages in this type of firming.

1. When fish fillets are dipped in strong brine, the proteins in the outer surface of the fillet
gets coagulated due to which firming takes place.
2. The layer of salted out proteins insulates the muscle tissues from the negative influence
of outside factors.
3. It favours the retention and preservation of natural extractives and flavour compounds.
4. It minimized drip loss.

Effect of salt concentration in brine and brining time

The treatment of fish fillets with brine is a precision operation. When the salt
concentration is less than 5% proteins get solubilized and lost in salt solution. If it is more than
15%, tissue proteins coagulate due to “salting out” of the proteins and the texture becomes tough.
If the duration of dip treatment is more, the product becomes salty. Therefore, brining treatment
should be done in right concentration for a brief period of time.

Method of brining: Before the treatment with brine, the fillets are washed properly,
because brine treatment on a clean surface is more effective. Salt concentration in brine should
be 10-12% (sp. gr. 1.07 to 1.09). Brining temperature should be less than 15OC. In dip treatment
the brining time is 1 to 2 min. However, there is scope to change the salt concentration and
duration of time for treatment depending on species of fish. The brine should be changed after
every dip treatment. Addition of antimicrobials such as sodium benzoate, benzoic acid, parabens,
sodium nitrite and antioxidants like ascorbic acid can be added to brine in order to maximize its
effectiveness. However in fatty fish bringing promotes fat oxidation in it and hence the
results are not encouraging.

Effect of salt concentration in brine and brining time

The treatment of fish fillets with brine is a precision operation. When the salt
concentration is less than 5% proteins get solubilized and lost in salt solution. If it is more than
15%, tissue proteins coagulate due to “salting out” of the proteins and the texture becomes tough.
If the duration of dip treatment is more, the product becomes salty. Therefore, brining treatment
should be done in right concentration for a brief period of time.

Method of brining: Before the treatment with brine, the fillets are washed properly,
because brine treatment on a clean surface is more effective. Salt concentration in brine should
be 10-12% (sp. gr. 1.07 to 1.09). Brining temperature should be less than 15OC. In dip treatment
the brining time is 1 to 2 min. However, there is scope to change the salt concentration and

61
duration of time for treatment depending on species of fish. The brine should be changed after
every dip treatment. Addition of antimicrobials such as sodium benzoate, benzoic acid, parabens,
sodium nitrite and antioxidants like ascorbic acid can be added to brine in order to maximize its
effectiveness. However in fatty fish bringing promotes fat oxidation in it and hence the
results are not encouraging.

3.4.2. Advantages of brine treatment

1. Treatment with brine destroys the microorganisms present on the fish fillet to a great
extent. However, salt should not contain Ca++ and Mg++ impurities which are responsible
for salt odour and bitter taste.
2. Prevention of belly bursting by brine treatment: During chilled storage and during
freezing and thawing, belly bursting is observed in sardines (poor appearance, viscera
comes out) which is a major problem in small fish such as sardine. If sardine is dipped in
15% brine for 30 min and then frozen, the problem of belly bursting can be minimized.
By this method, due to coagulation of proteins the belly wall becomes tough. However
the treatment promotes oxidation of fat and hence the shelf life in frozen storage gets
reduced.

Phosphate treatment : PolyPO4 treatment minimizes drip loss in frozen fish and fishery
products. PolyPO4 (Polyphosphate) treatment may be given separately or along with brine
treatment. In cod fillets drip treatment in 0.0844 < (4%) polyPO4 solution minimized significant
amount of drip loss.
Role of Polyphosphate in minimizing drip loss :
In frozen fish, water holding capacity of tissues comes down due aggregation and protein
denaturation and hence results in increased drip loss. The important reasons for drip loss are
given below.

1. Freezing changes ionic environment of fish muscle.

2. Reduction in pH due to production of lactic acid.
3. Degradation of nucleotide which contains active phosphorus.
4. Formation of actomyosin from actin and myosin.

The role of polyphosphate in minimizing driploss and preservation of quality of fish is explained
as follows.

1. Balanced action of tripoly PO4, organic phosphate such as ATP and other phosphates
leads to break down of the complex protein actomyosin to constituent proteins actin and
myosin resulting in increased water holding capacity of muscle.
2. Cell fluids of fish (salted) dipped in brine require -22OC for complete freezing due to
increase in salt concentration. Therefore liquid present in such cell fluids remain unfrozen
and atmospheric oxygen dissolve in such a liquid and penetrates inside the tissues. This
accelerates oxidation process. However, if 12% tripolyPO4 is mixed with brine, the

62
 freezing point of tissue fluids will only be -3OC. Therefore proportion of unfrozen liquid
will be greatly reduced and also concomitant oxidation. This is very much useful in fatty
fishes.
3. Polyphosphate acts as a chelating agent and chelate proxidant metal ions, thereby
minimizing catatytic action of metal ions in fat oxidation.

3.4.3. Use of Polyphosphate in freezing of fish

Dip treatment in phosphate solution prior to freezing gives better results. In prawns, in addition
to dip treatment in phosphate solution, it may be mixed with glaze water.

Quantity of polyPO4 to be used

Fairly high proportion of phosphate ion is required for breakdown of complex acto-l-myosin into
its constituent proteins. Among various phosphates, tripolyPO4 and pyropolyPO4 gives the best
results.

Use of tripolyphosphate solution of 0.16 M ionic strength (8%) and 4.5% pyropolyPO4 along
with salt to increase the ionic concentration to 0.4 M and 0.35 M respectively increases the water
holding capacity of muscle tissues effectively. Among the above, tripoly phosphate is extensively
used but its effect depends on the species of fish. Though tripolyPO4 solution and brine can be
used separately, fish and fillets are treated with pyropolyPO4 and brine together very good effect
was observed.

Polyphosphate should be used in such a way that it is absorbed quickly into fish muscle. It has
been found that the enzymes and digestive juices convert the polyPO4 in to orthoPO4 in the body
and hence it does not pose any health hazard. However it is better to take proper care while using
depending on its desired effect on the product.

Antioxidant treatment

Rancidity is the commonly observed storage problem in fatty fish. To minimize the problem of
rancidity, antioxidants which are commonly used in edible oils and fats have to be used in fish.
Rancidity in oils and fats is controlled by following methods.

1. Removal of pro-oxidant chemical compounds or inactivation of these factors.

2. Packaging of fish so that oxygen is prevented from coming into contact with fish surface
and subsequent penetration inside the tissues.
3. Before or after freezing fish may be treated with antioxidants. Enzymes responsible for
fat oxidation may be inhibited.

Phenolic antioxidants, even though very effective, they are not soluble in water. Some of the
methods used for application of antioxidants for frozen fish and fillets are as follows.

63
 1. Water soluble antioxidants such as ascorbic acid penetrates the tissues of fish through
tissue fluids and helps in arresting oxidation process. It is effective against salmon, trout,
herring and tuna during frozen storage. It is also effective against colour change in
pomfrets. Ascorbic acid also acts as a synergist when used along with BHA, tocopherol
and hydroquinone. Ascorbic acid in glaze increases the shelf life of mackerel and pomfret
during chilled and frozen storage.
2. Glaze containing ascorbic acid and citric acid mixture is effective against fatty fish and
onset of rancidity can be delayed for a long time.
3. Oil soluble antioxidants are useful in minimizing rancidity in fatty fishes. This is because
the oil of fish is on the surface and the antioxidants dissolve quite easily into such oil and
penetrates into the tissues. The antioxidant power of BHA and Ascorbic acid double
when used as a mixture.

3.4.4. Methods of using antioxidants

1. Dip treatment prior to freezing

2. In glaze water

Water soluble antioxidants can be easily dissolved in water and can be directly used. However,
oil soluble antioxidants are first emulsified in small quantity of oil in water type and used. When
gallates are used there should be no contact with Fe++ and pro oxidant metal ions may be bound
by chelating agents and make them unavailable in oxidation reactions. Nitric acid, phosphoric
acid, sodium phosphate citrates, tartarates and ethylene diamine tetra acetic acid (EDTA) are
some of the important chelating agents. These may be used separately or along with anti
oxidants.
Commonly used antioxidants
Antioxidants are of 2 types 1) Water soluble and 2) Oil soluble.
Antioxidants used in fish processing are as follows

Compound Nature Max. Percentage used

1) BHA Oil 0.02

2) Salts of gallic acid
a) Propyl gallate Water 0.01
b) Octyl gallate Oil 0.01
c)Do decyl gallute Oil 0.01
3) Nor-dihydrogui acetic acid Oil 0.01
4) Ascorbic acid Water 0.25-2
5) Sodium ascorbate Water 0.25-2
6) Sodium sulphide Water 0.25-2
7) Tocopherols Oil 0.03

64
 Amino acids (cystine, Water 0.03
8)
histidine, tryptophan)
Synergist (citric Water 0.03
9)
acid,sorbitol)

As antioxidants break the chain of free radical formation, oxidation is prevented. However, this
is easy in the case of almost pure oils but in the case of fish it is quite difficult because the action
of antioxidant depend on several factors such as

1. Property of oil in the tissues.

2. Penetration of antioxidants into tissues .
3. Presence of Pro-oxidant materials.

Therefore some antioxidants are effective only against certain species of fish. Some problems
associated with the use of antioxidants in fish are

1. Determination of an antioxidant for a particular species of fish.

2. Medium and method of application of the antioxidant.

65
Chapter 5: Calculation of freezer refrigeration load

3.5.1. Calculation of freezer refrigeration load

The individual items to be taken into account in a refrigeration load calculation depend on the
type of freezer. It would be impossible to include all the eventualities in one sample calculation;
therefore, a relatively simple one is given below for a HPF and some notes have been added to
help with other freezer calculations.

Load calculation

I. Number of freezer

20 t/day = 20,000 kg/day

20,000 ¸ 10 = 2,000 blocks/day

24 ¸ 2 = 12 cycles/day

2,000 ¸ 12 = 167 blocks/cycle

II. Fish load

20,000 ¸ 24 = 833 kg/h

Enthalpy at –30oC = 4.6 kcal/kg

Change in enthalpy = 81.3 kcal/kg

An approximate figure can also be calculated by using the following values:

a) Specific heat of fish above freezing, 0.9 kcal/kgoC

b) Latent heat of the fish, 60 kcal/k

c) Specific heat of fish below 0oC, 0.4 kcal/kgoC

Using these values, the above calculation for fish refrigeration load would be:

Heat to remove on cooling to 0oC

833 x 0.9 x 10 = 7497 kcal/h

Latent heat to remove 833 x 60 = 49980 kcal/h

66
Heat to remove on cooling to –30oC

833 x 0.4 x 30 = 9996 kcal/h

Total heat to remove from fish = 67473kcal/h

Total refrigeration requirement with allowances:

Add 30% = 142681 kcal/h

The calculation of fish load gives refrigeration requirement to freeze the fish only. Depending on
the type of freezer used, other heat loads such as fan heat, pump heat from circulating pump, heat
leak through freezer insulation, heat load due to pallets, trays, trolleys, defrost procedure, air
infiltration,internal lighting etc. have to be taken into account and added to this value to
determine the total refrigeration requirement.

3.5.1.1. Freezing time

Following are the Variables which affect freezing time

1. Freezer type
2. Freezer operating temperature
3. Refrigeration system and operating condition
4. Air speed in an air blast freezer
5. Product temperature
6. Product thickness
7. Product shape
8. Product contact area and density
9. Product packaging
10. Species of fish

Freezer type: The type of freezer will greatly influence the freezing time. For example, due to
improved surface heat transfer, a product will normally freeze faster in an immersion freezer than
in an air blast freezer operating at the same temperature.

Operating temperature: The colder the freezer, the faster the fish will freeze.

Air speed in blast freezers: Freezing time is reduced as the air speed increased.

Product before freezing: The warmer the product, the longer it will take to freeze. Fish should
therefore be kept chilled (+4oC) before freezing both to maintain quality and reduce freezing
time and refrigeration requirement.

Product thickness: The thicker the product, the longer is the freezing time.

67
Product shape: The shape of a fish or package can have a considerable effect on its freezing
time and this is dependent on the ratio of surface area to volume.

Product contact area and density In a plate freezer, poor contact between product and plate
results in increased freezing time.

Product packaging: The method of wrapping and the type and thickness of the wrapping
material can greatly influence the freezing time of a product. Air trapped between wrapper and
product has a greater influence on the freezing time than the resistance of the wrapping material
itself.

Species of fish The higher the oil content of the fish the lower is the water content. Most of the
heat extracted during freezing is to change the water to ice; therefore, if there is less water, then
less heat will require to be extracted to freeze the fish. Correspondingly freezing time will be
less.

3.5.1.2. Calculation of freezing times

The more general from of Plank’s equation for calculation freezing time is:

L PD RD2

Freezing time = -----{------ + ------}

V D fk

Where

L = Heat to be extracted between the initial freezing point and final temperature (kcal/kg).

V = Specific volume of fish (m3/kg)

D = Temperature difference between the initial freezing point of the

fish and the refrigerating medium (oC)

D = Thickness of product in direction or prevailing heat transfer (m)

f = Surface coefficient of heat transfer (including effect of

packaging) (kcal/h.m oC)

k = Thermal conductivity of frozen fish (kcal/h moC)

68
P and R= Constants which depend on shape

Values for shape constants P and R

Shape P R

Sphere 0.167 0.042

Infinite Cylinder 0.167 0.042

Infinite Slab 0.500 0.250

Calculated freezing time – Example 1: What is the freezing time if all other conditions remain
the same but the operating temperature is –25oC.

Fish freezers at about –1oC, therefore in the measured freezing time, the effective temperature
difference is 34ºC (the difference between –35oC and –1oC). The effective temperature difference
for the freezing time required is 24ºC (the difference between 25oC and –1oC). Freezing time is
inversely proportional to the temperature difference, therefore the freezing time with an
operating temperature of –25oC will be longer than for a temperature of –35oC and can be
calculated as follows:

200 x 34 % 24 = 283 min or 4h 43min

Calculated freezing time – Example 2 What is the approximate freezing time if all other
conditions remain the same and the block thickness is reduced to 75mm?

Freezing time is directly proportional to the square of the thickness since in this case the surface
heat transfer coefficient is high and the factor relating to the thickness of the block, PD/f, will be
small. The new freezing time will therefore be calculated as follows:

200 x 752 ¸ 1002 = 200 x 5625 ¸ 10000 = 112 min (1h 52 min)

3.5.1.3. Coding, Packing and Storage

A code slip bearing the marking of the numerical code of the processor, abbreviation of the name
and type of products, year, month and date of production shall be embedded on the frozen block.
In the case of individually Quick frozen (IQF) packs the code slips shall be placed in the primary
containers.

An illustration of the coding is given below:

69
520 FSPD

11A10

Where in the above illustration

520 – the numerical code of the processor/ exporter

FS – Name of the product (Frozen shrimps)

PD – Type of the product (Peeled and Deveined)

11 – Year of processing (here 2011)

A – Month or processing (here January)

10 – Date of processing (here tenth day of the month)

The following abbreviations shall be used for the months of the year:

January - A

February - B

March - C

April - D

May - E

June - F

July - G

August - H

September - J

October - K

November - L

December - M

The frozen products shall be packed in primary container other wise known as wax coated board
or duplex canton and such primary containers shall be packed in seaworthy fibre board cardboard

70
cartons of 5 ply minimum or any other better type of cartons. In the case of Individually Quick
Frozen products the pieces shall be arranged/wrapped in moisture proof film. As per the specific
requirements of the buyer the packing of frozen fish and Fishery products without primary
container/Duplex cartons shall be permitted. In such cases the products shall be placed in a
master poly bag and placed inside the master carton. The frozen material shall be stored in a
room maintained at or below a temperature of minus 18o C.

71
Chapter 6: Freeze drying fish

3.6.1. Introduction

Heat is used to dry foods or concentrates liquids by boiling to remove water and therefore
preserves the food by reduction in water activity. However, heat causes a loss of sensory and
nutritional qualities. In freeze drying a similar preservative effect is achieved by reduction in
water activity without heating the food and sensory and nutritional qualities are consequently
better retained. However both, operations (freezing & drying) are slower than conventional
dehydration. Freeze drying has become an important commercial operation to dry expensive
foods which have delicate aroma or texture (for example instant coffee, mushrooms, fruit juices
and sea foods).

3.6.2. Theory of freeze drying

The boiling point of water depends on the overhead atmospheric pressure. At normal
atmospheric pressure of 1 bar (i.e., 760 mg of Hg) it boils at 100ºC. However, if it is held in a
sealed container and overhead atmospheric pressure is lowered than one by using vacuum
pumps, then it boils at less than 100ºC. It is observed that the lower the pressure (or the better the
vacuum) the lower is the boiling point of water but the energy needed for evaporation of the
same quantity of water is more. The pressure, the corresponding boiling point of water and latent
heat of evaporation are presented in the following table.

Pressure (bar) Boiling Point of Water Latent heat of

O
C vaporization (kj/kg) of water

1 100 2257

0.5 81 2305

0.2 60 2358

0.1 46 2392

0.01 7 2485

At OOC water exists an ice (solid phase) and if the overhead pressure is sufficiently low,
solid ice will directly sublime as water vapour without going into liquid phase. To understand
the phenomenon more clearly it is essential to refer to the phase diagram of water.

Pressure units conversion table :

72
 Torr mm Hg Pascal

1 1 133.3

10-1 0.1 13.33

10-2 0.01 1.333

From the phase diagram it is clear that if the temperature of water and pressure is below the
triple point it exists as ice and it will directly pass into vapour phase.

If the overhead pressure of a food is held below 4.58 Tor (610.5 Pa) and the food is
frozen, when the frozen food is heated solid ice sublimes directly to vapour phase without
melting. Water vapour is continuously removed from the food by keeping the pressure in the
freeze drier cabinet below the vapour pressure at the surface of ice by removing the water vapour
with a vacuum pump and condensing it on refrigeration coils or plate or drum. As the drying
proceeds, the sublimation process continues as long as latent heat of sublimation is
provided/supplied. The latent heat of sublimation is either conducted through the food to the
sublimation front or produced in the food by microwaves. The pressure, sublimation temperature
in the frozen state and corresponding latent heat of sublimation are presented below.

In freeze drying, foods are dried in two stages, first by sublimation to approximately 15%
moisture content (on wet weight basis) and then by evaporative drying of unfrozen water to 2%
of moisture content (W.W.B). Desorption is achieved by raising the temperature in the drier to
near ambient temperature while retaining the low pressure.

The rate of drying depends mostly on the resistance of the food to heat transfer and on the
resistance to vapour flow.

3.6.3. Principle of the equipment

Effectiveness of a freezes drier depends on rate of heat transfer to the sublimation front and rate
of mass transfer.There are three methods of heat transfer.

Heat transfer through the frozen layer. The rate of heat transfer depends on the thickness and
thermal conductivity of frozen layer. As the drying proceeds, the thickness of ice is reduced and
the rate of heat transfer increases.
Heat transfer through dried layer. The rate of heat transfer to sublimation front depends on the
thickness of the dried layer and the surface area of food, thermal conductivity of dry layer and
the temperature difference between the surface of the food and ice front. The dried layer has very

73
low thermal conductivity and therefore offers high resistance to heat flow. As drying proceeds,
this layer becomes thicker and the resistance increases.
Heating by microwaves : Heat is generated at the ice front and the rate of heat transfer is not
influenced by thermal conductivity of ice or dry matter or the thickness of dry layer. However it
is difficulty to control microwave heating.

Rate of mass transfer: It depends on the water vapour pressure gradient inside the drying
chamber which is controlled by

1. The pressure in drying chamber

2. Temperature of the vapour condenser – both should be as low as possible.
3. Temperature of ice at the sublimation front and it should be on high as possible (at OoC)
without melting.

Equipment : freeze drier consists of a vacuum chamber containing trays to hold food during
drying, source of heat to supply latent heat of sublimation, refrigeration coils to condense vapour
with automatic devices to keep maximum area of the coil free of ice for vapour condensations
and vacuum pump to remove non expensible vapour.
Types of equipments :

1. Accelerated freeze driers : Food is held between expanded metal mesh for heat transfer.
2. Radiation freeze driers : In paired radiations from radiant are used. Heating is more
uniform. Close contact between food and the heat is not necessary; float trays can be used
and cheaper. Easier to clean.
3. Microwave and dielectric freeze driers

3.6.4. Freeze drying process

Freeze drying cycle consists of 3 stages.

1. Freezing.
2. Primary drying to remove water ice.
3. Secondary drying to remove bound water molecules from the primary dried materials.

1) Freezing : Freezing the product temperature is reduced to -50OC at one atmospheric pressure
to ensure that it is below the eutectic point.

2) Primary drying : This is a dynamic process involving transits of ice phase to vapour phase
directly. This leaves the solid matrix formed in the freezing phase but without the ice crystals.
During drying, an interface is set up which comes from the exposed surface to vacuum system to
the surface where heat source is in contact or moves deeper in radiant heat. At this interface,
actual drying takes place. There are number of factors which will indicate the completions of
primary drying process.

74
1) Pressure stability : During drying, pressure changes slowly until it stabilizes at an ultimate
low level. The process is considered as complete where the constant pressure is maintained for 1
to 2 hours.

2) Product temperature rise. When heat energy is not required for sublimation, the overall
product temperature will rise until equal or close to the chamber temperature. This can be
detected with a temperature probe and displayed.

3) Pressure rise test. If an isolation valve is available, it will be possible to isolate the drying
chamber. If the material is not dry, then the pressure will rise quickly (due to vapour pressure
of the product).

Secondary Drying: This is part of the process where remaining small proportion of “locked in”
water is removed ( Usually 2.5% by volume). This is not always considered necessary and
depends on the product.The locked in or bound water is removed form the products by
supplying additional heat energy to the system. This is achieved either by increasing the
temperature or by improving the thermal contact between the product and the existing of heat
source.

Effect of freeze drying on food : Freeze dried foods have high retention of sensory and
nutritional qualities and shelf life of more than 12 months when correctly packed. Aroma
retention is 80-100%. Shrinkage is very little and texture is well maintained. Allows rapid and
full rehydration but the product is fragile and require protection from mechanical pressure.
However, the open porous structure of the food may allow oxygen to enter and cause oxidative
deterioration of lipids and subsequent browning. Changes in thiamine and ascorbic acid content
during freeze drying are moderate and there are negligible losses of other vitamins. Therefore
vacuum or inert gas packaging is required.

75
Unit 4: Thawing of fish

Chapter 1: Methods of fish thawing

4.1.1 Introduction

Thawing is a process whereby heat is introduced into the frozen product. In order to raise its
temperature above freezing point, the rate of heat input required to thaw 1 ton of fish per hour is
approximately 85 KW. Thawers or methods of thawing can be divided into two broad classes.
1) Those which depend only on the conduction of heat through the thawing material.
E.g. Thawing in air, thawing in water and thawing between heated plates.
2) Those which do not depend on the conduction of heat through the thawing material.
E.g. Dielectric thawing and electric resistance thawing.
There are also other methods such at infra red and microwave heating, but they are not of
interest for thawing on a large scale

4.1.1.1 Thawing in air

a) Natural convection:

It is sometime convenient to thaw at room condition, if conditions are suitable. Temperatures

above 18ºC should not be used. Lot of space and labour is required for thawing on a large scale
by this method and the introduction of humidification to prevent slight drying of the fish is
difficult.
b) Air blast:
In this method, a current of air is directed on the frozen block and the increased air
velocity reduces thawing time. The rate of thawing is mainly dependent on the resistances of the
fish itself to the flow of heat. At the air velocity of 6m/sec, there is very little resistance, which
is optimum. In modern air-blast thawers with the recirculation of humid air at 20oC( the
maximum, temperature that should be employed), the 10cm block of frozen fish requires about 5
hours for thawing before they can be filleted. The fish can be separated easily after about 3
hours, which reduces the thawing time to about 4 hours and gives more even thawing. Complete
humidification of the air is necessary in order to prevent drying of the fish and to achieve ideal
condition for heat transfer. Weight losses in the fish normally do not exceed 1%. The problem
of cleaning is greater with the air blast thawers when compared with most other types. Two
types of air blast thawers, i) continuous ii) batch.

i) Continuous:

Continuous air-blast thawers have been built in two designs. Cross flow and parallel
flow. In the cross flow design, the direction of air flow is across the conveyor at right angle.
Typically there are five conveyors, one above the other. The fish travels horizontally along the
top conveyor until when it reaches the end, it falls to the conveyor below which is traveling in
the opposite direction and so on. In order to achieve high velocity, capacity of the fan should be

76
high with this arrangement and use of fan power is more than 30 KW for a unit fish. Thus a
large portion of the heat for thawing is obtained from the fan.

In the parallel flow thawers, the air travels in the same direction as the conveyor. With
humid air, most of the heat transfer to the fish takes place by condensation at water vapour and
an air temperature drop at 300C can be tolerated. The fan capacity is relatively low and
therefore, the fan power is about 7.5 KW for the 1 ton/hour unit and the face area of the heater
and humidifier in conveyor is, in a single line occupying a greater floor area, than a cross flow
thawers. In the other, the conveyor is divided into three sections, one above the other.

ii) Batch:

The largest model of batch air-blast thawers has five compartments each holding 1 ton of
10cm blocks of whole fish on two trolleys with suitably- spaced shelves, giving an output at
about 1 ton/hour. Air flow is in series through the five compartments in a line. Periodic reversal
at air flow can be employed to even out differences between compartments. The thawers can
also be used on a semi continuous basis by inserting and withdrawing trolleys at intervals of 1
hours. More labour for loading and unloading is required than for continuous thawers, but the
batch thawer is simple and compact. The 20 KW fan heaters and humidifier are located above
the fish compartments. Batch thawers with small fans giving moderate velocities, in which
blocks of fish are placed on racks or shelves, have been used for thawing frozen fish for over
periods of up to 10 hours.

4.1.1.2 Water thawing

a) Immersion:

The thawing of blocks of whole in water has been shown to give good results, but has
been used only to a limited extent for thawing on a large scale. The thawed product can be as
high in overall quality and suitable for processing. Pump capacities for water thawers need not
be great, because the water velocity required for good heat transfer is 0.5 cm/sec, giving thawing
rates equal to those for air-blast. It is probably necessary, to use somewhat higher velocities,
perhaps 2cm/sec or more, in order to give a good degree of mixing of the water, preventing
stratification and to provide reasonably quick response to the heater, particularly for warm up
with initially cold water. Too high velocities induce turbulence, causing foaming at the surface
of the water and possibly lead to water logging. The temperature of the water should conserve
heat, it is better to change the water once or twice in 24 hour so as to limit the degree of
contamination of the water.

b) Spray:

In this, larger thawers are used, in which heated water is sprayed directly on the fish. The
blocks of fish travel on a conveyor, which is made up of a series of open mesh, wire baskets in a
housing ventilated by a small fan. The water is recirculated through a special filter and cleaner
section. The thawing time is claimed to be at least as short as in air, blast thawing. Hygienic

77
condition is maintained in the thawers because of the working action of the sprays of clean water.
It takes about 5 hours for the 10cm block of whole fish to be thawed completely, in a thawer with
are output of 1ton/hour.

4.1.1.3 Thawing between heated plates

This type of thawer is similar to a plate freezer except that warm fluid, normally at 20ºC is
circulated throughout the (metal) plate. The product is held in contact with the plates throughout
the thawing period which is about 8 hours for complete thawing of the 10cm block of whole fish.
Thawing is more even than on air blast thawing, with the result that the fish can be filleted
sooner, typically 7 hours after an initial period at 4 hours in the thawer as opposed to 12 hours in
air-blast thawing. Little power is required excluding the heat for thawing, the plant is compact
and robust

a) Dielectric thawing:

In this method, the frozen material is placed in a high frequency electrical field between two flat
metal plates. An alternating voltage at 5,000 V and 40MHZ is applied to the plates and hence heat
is produced in the electrically charged particles in the food. The plates are fed from a generator
and suitably shielded to confirm the field. Unless the food is homogenous and regular in shape,
there is possibility for uneven heating so that the parts of the material are cooked while other
parts remain frozen. This can be minimized in block at fish continuously conveying them
through the field and restricting the power density according to the degree of thawing. Thus, a
typical industrial thawer for blocks at fish includes a conveyor belt and six generators each with
a rating of 20KW giving a nominal output of 1 ton /hr with 10 cm blocks of whole fish. Thawing
is very rapid so that the total length of conveyor has been much shorter than that for continuous
air-blast thawers. The thawing time for blocks of large white fish is about 80 min, and for blocks
of small fatty fish is about 20min.

b) Dielectric and water:

The dielectric thawer has a thermal efficiency of about 70%. Cooling water for the generators
absorbs the waste heat. It has been suggested that heated water be used for the thawing either
before or after the dielectric thawer, a hybrid system, in order to increase the output and thermal
efficiency and reduce the risk of overheating. Since the heat conduction is rapid during the early
stages, the 10 cm block of whole fish can be 30% thawed in less than 30min in water at 15oC but
the complete thawing takes up to 80 min at the temperature at 15oC.

c) Electric resistance thawing:

In this method, fish can be heated by placing it between two electrodes used applying a low
voltage so that a current flows through. Like dielectric heating, it is difficult to achieve even
heating unless the item is homogenous and regular in shape. It is convenient to partially heat the
fish by a heat conduction method such as immersion in circulating tap water and thawing by heat
conduction is rapid during the early stage. Usually the time taken for water immersion and
78
electric heating should be roughly equal. The total thawing time is between 20 and 60 min
depending on the type at blocks, at least twice as fast as thawing in air or water. A complete
batch thawer of this type has plates and blocks of fish arranged in layers one above the other in
the same way as a horizontal plate freezer. The plates are heated electrically or by passing worm
fluid through them during the early stage at thawing when heat conduction is employed and act
as electrodes during the stage of electric resistance thawing.

d) Vacuum thawing

Vacuum thawers consist of airtight chambers in to which the fish is loaded using trolleys. The
temperature of the water vapour is typically between 18oC and 20oc and the vapour condenses on
the cold surface of the fish where the latent heat of vaporization is absorbed by the fish meat
transfer to the product is much more rapid than the previous method of thawing. Equivalent
thawing fishes may be reduced from 20hr in air to less than 1 hr in a vacuum thawing unit and
the water usage at such units is low. However, care must be taken that gases released from the
fish as thawing proceed do not cause rupturing at flesh.

The advantages of this method of defrosting are that with the exception at a vacuum pump there
are no moving parts, hence the possibility of mechanical breakdown is reduced compared to the
previous methods. Also the vapour is distilled from water in the tank it condenses on to the fish
in a pure state, so there is no risk of contamination from bacteria which may accumulate within
the system.

4.1.1.4 Choice of method

Many factors should be considered when choosing between the various methods. The choice
depends largely on the type of product and the form in which it was frozen. In some uses, it is
difficult to provide sufficient shielding in the dielectric thawer. The air-blast thawer might be at a
disadvantage in the amount of cleaning required to keep bacterial counts within the statutory
limits. All the main methods described above can produce satisfactory results with blocks of
whole fish. Both heat conduction and electric methods fail with large fish. The electric methods
give more uniform thawing with compact block at small fish. 'Continuous thawers’ would appear
to have advantage over batch thawers for blocks at whole fish. For blocks of fillets, water
thawing is unsuitable and block thickness probably should not exceed 5 cm for thawing in humid
air blast and between heated plates. The electric methods give good results with blocks of fillets.
Block upto 10cm thick have been thawed successfully dielectric thawer using only three
generators. Fillets, unprotected by the skin of the fish, are more susceptible than whole fish to
deterioration on thawing, particularly shrinkage, discolouration and dehydration. Pronounced
thaw-rigor accompanied by shrinkage and loss of fluid sometimes can be produced when
thawing is rapid.

79
Unit 5: Quality changes during frozen storage

● Chapter 1: Quality changes in fish during freezing & frozen storage-physical

changes

5.1.1 Quality changes in fish during freezing

The physical changes which occur during freezing & storage of frozen products comprise
crystallization of ice with expansion of the volume, and desiccation starting from the surface of
the frozen fish.

5.1.1.1 Ice formation

The crystallization of a ice is initiated when the temperature of the fish is lowered about
0
–1 C. At the same time, concentration of various inorganic salts and organic components present
in the fluid of the fish occurs and consequently the freezing point falls. The temperature of the
fish must be lowered further before additional water freezes. There is also an increase in the
volume of the fish when the water is converted to ice. At -30C about 70% of the water is frozen.
At -50C, about 85% is frozen; at-250C about 95% and at -500C to -600C, almost all the water in
the fish is frozen. The larger part of the water consequently freezes between -10C and -50C and it
is the rate of cooling this temperature interval determines the size of the ice crystals. The
preserving effect of freezing however is due to the extent of reduction in the rate of chemical and
biological processes, when the temperature of the fish is lowered. The crystallization or freezing
of the water has certain un favorable effects. Rapid freezing, results in small ice crystals, and the
quality of quick frozen fish may be practically equivalent to that of fresh unfrozen fish.

Microscopic examination of frozen fish reveals that the changes occurring in freezing and
thawing are caused rather by irreversible changes in the colloidal state of the proteins induced by
the increased concentration of dehydrates in the unfrozen part of the fluid in the fish
tissue. Small ice crystals are certainly a feature of quick freezing whereas large ice crystals can
be the result not only of slow freezing but also of considerable post mortem changes in the tissue
prior to freezing. Histological investigations on frozen fish revealed that when freezing fish with
considerable post mortem changes, the ice crystals destroy the sarcolemma more easily and grow
large sizes causing more histological changes to the tissue. The rate of freezing therefore has
been considered of vital importance for obtaining a high quality product. The DNA content in
the drip also varied with rate of freezing. With more rapid freezing, there is a drop in DNA
content in the expressible fluid. In ultra rapid freezing, however, there is a rise in expressible
DNA, but probably due to different kind of cell damage.

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 5.1.1.2 Desiccation

Desiccation from the surface of the frozen fish takes place during freezing and storage
because there is always transport of water vapour from the fish to the evaporator or cooling
coils. The greater the temperature difference between fish and evaporator, and greater the
velocity of the circulating air, the more moisture will be transported and the more frost will be
formed resulting in the decreased efficacy of the evaporator. The loss of water from the product
and the formation of snow on the evaporator coils are greater at high storage temperature,
because the warmer air will take up more moisture. Change in fluctuation in storage temperature
influences the desiccation, loss of weight and quality of the fish contributes to a poor appearance
or results in “freezer burn”. The critical limit of dehydration or rate of weight loss of frozen fish
in cold storage is 50g/m2/ 24h.

Medium sized white fish stored in a rather dry room lost 7, 3.5 & 1.5% in weight per
month at -9, -21 & -29oC , respectively. The air surrounding the frozen product is usually at a
lower moisture vapour pressure than the product and therefore acts as a sponge in removing
moisture from the product. The loss of moisture results in the dehydration of the flesh to a point
where chalky and fibrous texture develops, discolouration takes place and off flavours develops.
The phenomenon is also know as “Freeze burn”. When frozen fish is in contact with air,
oxidation of fat or oil in the fish takes place and these results in “rusting” or discolouration of the
flesh and development of rancid odours and flavours.

If the desiccation is pronounced, the fish surface may become dry and fibrous. In some
cases the skin may change the colour, several other factors influence the loss are weight, the
kind of wrapping, it’s sealing and moisture transmission characteristics. The rate of moisture
loss or weight loss or water vapour transmission rate (WVTR) from fish in cold storage is
expressed in g/m2/day. The denaturation of the protein or reduced protein extractability is also
due to the electrostatic interaction between protein and fatty acid. Freezing increases toughness
of fish muscle, which proceeds progressively during subsequent storage. Toughness can not be
measured in terms of percentage of soluble protein in the cold stored fish. The proportion of fish
muscle protein soluble in 5% Nacl solution was not correlated with the development of
toughness during cold storage at or below -200C. Assessment by a taste panel is time consuming
and inaccurate. Since fish muscle fibres are extremely short and interspaces. Instruments such
as penetrometers have limited usefulness. But by using a new method of treating the fish muscle
with 1% formaldehyde, it was found that the unfrozen fish muscle fibres were broken up & there
by reducing the light transmission. Frozen fish showed a higher proportion of intact fibres,
letting through more light.

Toughness:

Toughness or changes in texture can be due to denaturation of proteins. The denaturation may be
due to the concentration of inorganic salts which results in damage to the flesh proteins or due to
lipid hydrolysis and myoglobin in the cells, are the main colouring pigments in fish flesh. The
dark parts of the muscles contain much more pigment than the light or white muscle.

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 5.1.1.3 Discolouration

In freshly caught fish the blood has a brick red color of oxy hemoglobin. This color gradually
changes to the red brown or brown color of met hemoglobin. Myoglobin suffers the same kind of
changes. Frozen tuna and sword fish may exhibit green and brown discolouration on looking.
Uncooked fish meat contains three derivatives of myoglobin. The pigment responsible for the
pink colour in normal cooked meat of tuna is hemochrome, derived from the reaction of
myoglobin with non-heme constituents. Greening is due to pigments resulting from the oxidation
of hemochrome that occurs when the meat is unduly exposed to oxidative condition during and
after cooking. Greening of frozen sword fish may be related to take H2S produced by
putrefactive bacteria. Proper evisceration and removal of blood immediately after the catch
reduce the risk of discolouration. The undesirable discolouration in yellow fin tuna meat can be
averted, if the fish is frozen at full rigor, stored at a temperature of -230C to -270C, and defrosted
by still air at 100C.

a. Carotenoid pigments:

Some fishes like salmon, and some crustaceans, contain carotenoid pigments in the flesh. The
red color astaxanthin tends to fade away during frozen storage. In crustacean, like lobster,
pigment is limited to the surface of the meat, the changes during frozen storage lead to yellow
discolorations.

b. Fish oils:

The oils & fats naturally occurring in fish vary with species from colourless, as in herring
through yellow to red, as in salmon. Fat herring, with a layer of oil just under the skin, may
develop a yellow color during poor storage condition & which can be seen through the skin.
Such fish is said to have “rusted”.

c. Black spots:

Shrimp, lobster etc, develop so called black spot when stored fresh. Bruises and rough handling
increase the occurrence of this discolouration, which is caused by an enzyme, tyrosinase acting
on polyphenolic compounds, present in free amino acids like tryptophan,tyrosine and form black
'melanin' pigments.

Flavor changes:

Systematic studies of flavor changes in frozen fish are lacking. Freshly processed fish, frozen
and cold stored, is generally considered first to lost it’s characteristic fresh fish flavour,
becoming bland & insipid; there after it develops flavors variably described as “cold stored”
“Salt fishy’ etc.

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Break down of connective tissue:

During some occasion, fish fillets are produced in which the muscle flakes will separate when
the fish is filleted. This defect, often called “gaping” appears to result from a breakdown in the
connective tissue and associated components which hold the muscle cells together. Although the
main myofibrillar proteins, actin and myosin are believed to remain largely intact, many of the
cytoskeletal protein like connectin, nebulin and desmin are believed to be degraded. The problem
is, by and large, connected with freezing at sea and fillet production from frozen whole fish.

5.1.1.4 Fat oxidation during freezing

The changes in lipids are directly and indirectly responsible for the quality deterioration in frozen
seafoods. They involve lipolysis, lipid oxidation, and interactions of the products of these
processes with non-lipid components. The endogenous fish lipases are relatively resistant to low
temperatures and retain much of their activity in the frozen tissues. Some of them may even be
activated in the freezing process eg. by release from lysosomes. Fish lipids undergo two main
types of changes during storage of fish viz, hydrolytic changes and oxidative changes. These
changes result in rancidity.
Lipid hydrolysis
Lipases bring about hydrolysis of lipids producing free fatty acids and resulting in hydrolytic
rancidity. Free fatty acids trigger protein insolubilization and texture degradation in frozen stored
fish.
Lipid oxidation
It is a very series problem in fish, in view of the highly unsaturated nature of fish lipids. The
double bonds of unsaturated fatty acids are highly susceptible to oxidation and this leads to the
production of various carbonyls and other secondary oxidation products, which impart the
characteristic rancid off flavour to the product. These products, besides producing off flavour
reduce the shelf life and nutritional value of the product also. Some of them are toxic in nature.
These reactions are initiated by free radicals generated from unsaturated bonds, which start chain
reactions resulting in the production of various undesirable compounds like peroxides,
hydroperoxides, aldehydes, ketones etc. Finally, the free radicals form non-radical polymers,
which terminate the chain reaction.
Oxidised lipids interact with proteins reducing the nutritive value of the proteins considerably.
Melonaldehyde is one of the major oxidation products and estimation of this compound by
forming the thiobarbituric acid (TBA) complex is the accepted method for monitoring the extent
of lipid oxidation. In lipid oxidation, the first step leads to formation of hydroperoxides, which
are tasteless but can cause brown and yellow discolouration of the fish tissue. The degradation of
hydroperoxides gives rise to formation of aldehydes and ketones. These compounds have a
strong rancid flavour. Lipid oxidation primarily non enzymatic in nature, recently the
involvement of microsomal enzymes and lipoxygenase has been reported. This lipid oxidation
takes place in fishes having more than 2% of the lipids eg. fatty fishes.

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Factors affecting the oxidation

1. Content and composition of unsaturated fatty acid

2. Oxygen availability
3. Light radiation
4. pH
5. Temperature
6. Moisture content
7. Content of pro and anti oxidant.

5.1.1.5 Factors limiting storage life

Protein changes: Fish proteins undergo permanent change during freezing and cold storage and
the speed at which this denaturation occurs depends upon temperature. At temperatures not very
far below freezing point, -2oC for example, serious changes occur rapidly; even at –10oC, the
changes are so rapid than an initially good quality product can be spoilt within a few weeks. The
rate of deterioration due to protein denaturation, can be reduced by storage at as low a
temperature as possible.

5.1.1.6 Freeze Denaturation-I

Unfolding of molecules due to secondary reaction between the reactive groups of different
proteins and other components of fish muscle, leading to cross linking and formation of
aggregates

● The proteins lose part of their solubility and reduced enzyme activity
● As a result of these changes significant deterioration of the functional properties of the
fish meat may occur
● It is manifested by a decrease in water retention, gel forming ability and lipid emulsifying
capacity worsening of texture and the increased dryness of fish meat
● Fish that are stored for longer period becomes tough, chewy, rubber and fibrous
● Causative factors comprise the denaturing are

1. Catalytic effect of ice and inorganic salts

2. The binding of fatty acid

3. Lipid oxidation products

● Cross linking induced by formaldehyde generated in the muscle of some fish (Gadoid)
results in the formation of new covalent bonds in denatured products
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 ● Ice crystals disturb the hydrophobic interaction in protein. Increase in concentration of
salt in unfrozen food and water decrease the hydration of protein as the concentrated
inorganic ions compete for the water molecules
● They participate in freeze denaturation of protein by catalyzing the hydrolysis and auto
oxidation of lipids
● Free fatty acid decrease the solubility of myofibrillar protein and contribute to the
formation of aggregates by binding to proteins due to hydrophobic interaction
● Oxidised fatty acid and secondary products of autoxidation especially aldehydes are
much more damaging to protein solubility as they are able to form covalent bonds with
reactive protein groups.

TMAO —>TMA + formaldehyde (HCHO)

So aldehydes is very reactive compound bind to some groups in protein side chain and form intra
and intermolecular methylene bridges. By removing water soluble substrates and enzyme system
responsible for the degradation of TMAO the undesirable protein changes can effectively be
reduced.

5.1.1.7 Freeze Denaturation-II

Fat changes:

Fatty fish may become unpleasantly altered during cold storage but they can be protected to
some extent either by glazing or by packaging in plastic bags sealed under vacuum. These
oxidation changes take place more rapidly at higher temperatures and storage at a low
temperature is an effective means of slowing the rate of spoilage by this method.

Color changes:

The quality of fish is often judged by appearance, and color changes which are not otherwise
significant can result in fish being downgraded. The changes in the fish flesh which bring about
these color changes are also retarded at lower temperatures.

Dehydration changes:

Dehydration of the product is probably the major concern of the cold store operation and the rate
of drying can be linked with a number of factors in cold store design and operation.

When fish get badly dehydrated in cold storage, the surface becomes dry, opaque and spongy. As
time progresses, these conditions penetrate deeper into the fish until it becomes a fibrous, very
light material. Visible effects of severe dehydration on the surface of the fish are known by the
term “freezer burn”. This is an unfortunate choice of term since the effect is unlikely to result

85
from freezing in a properly designed freezer, and appears only after periods of storage in a cold
store.

Frozen fish may dry slowly in cold storage even under good operating conditions. This is un
desirable for reasons other than the obvious one that the product will lose weight. Drying also
accelerates denaturation of the protein and oxidation of the fat in the fish. Even totally
impervious wrappers used to protect the product do not give full protection if the cold store
operating conditions are favorable for desiccation within the pack. In-pack desiccation prevails
when there is some free space within the wrapper and temperature of the store fluctuates.

Weight loss during freezing will depend on: Type of freezer, freezing time, Type of product, Air
velocity and Freezer operating conditions.

Cold Store Weight Loss Depends on Temperature, Temperature fluctuation, Humidity, Heat
transfer, Air flow over the product, Radiation effects of lighting, The product, Shape and size of
the product and Type of wrapper.

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Chapter 2: Microbial growth at low temperature

5.2.1 Microbial growth at low temperature

Freezing of fish is done at – 400C and the frozen fish is further stored at – 180C. During
freezing, 80 to 90% of the Gram-negative bacteria die out and the residual bacteria cannot grow
in the temperature of frozen storage. So, during freezing preservation of fish, there is no
bacterial spoilage. But, before cooking, the frozen fish has to be thawed. During the thawing
process the residual bacteria, which are predominantly Gram positive, can cause spoilage of the
thawed fish. Hence, frozen fish will have to be thawed within the shortest possible time. The
relation between microbial growth and temperature is still considered as complex phenomena
governed by inter-related factors such as substrate composition, freezing rate, microbial type.
As the temperature falls, bacterial growth rate is reduced and the lag period is extended
until the minimum temperature limit is reached when growth ceases. Some microbes will cease
growing at 00C or even higher while others continue to grow below the freezing point of food.
Growth rate below 00C is very slow. The reported minimum growth of microorganism varying
from 100C to - 100C. But for practical cases, the lower limit of growth for bacteria, yeast or
moulds can take as -70C.

Most food stuffs contain low molecular weight compounds such as salts, sugars, etc.
which decrease the freezing point of water and lower the water activity. As more and more water
is converted to ice with the fall in freezing point, the water activity in the remaining unfrozen
water is reduced. Hence, microorganisms as they approache their minimum growth temperatures
are subjected to two factors mainly, i.e. (i) the concentration of the solutes in the unfrozen
water which destroys the osmotic equilibrium of cell membrane leading to metabolic
diffusion and (ii) strong inhibitory action due to low water activity. Additional stress
imposed on the cell prior to freezing causes changes in the enzyme activities of microbial
populations and reduces the efficiency of their metabolic pathways and makes them more
susceptible to low temperatures. This is of particular importance to food such as fish, which
is chilled prior to freezing. Hence, the ultimate effect of freezing is thought to be due to ice
crystal formation and solute concentration caused by the damage to the semi permeable
properties of the cell membrane. At temperature above -10°C, freezing occur only externally and
a cell that could make osmotic adjustments and escape death. On the other hand, below -10°C,
cell membrane fails to act as an osmotic barrier to the proliferation of ice already formed outside
the cell resulting in intracellular ice formation and death. Damage to DNA is also involved.

When a food product having a mixed flora is exposed to low temperature, a small change
in temperature leads to substantial variation in the relative growth rates of the micro-organisms
present. It has been conclusively proved that Gram-positive bacteria particularly Gram-positive
cocci are most resistant to deleterious action of freezing. Yeasts and molds that are able to grow
at relatively lower water activities are also a common component in frozen foods. Cells those are

87
not killed by freezing may be sub-lethally injured and such injury is reversible. The repaired
cells regain all the normal characteristics of the original cell within a few hours and this happens
in the lag period. Injured cells often exhibit increased nutritional needs, impaired membrane
permeability, reduced resistance to environmental stress (e.g. low pH) and increased sensitivity
to inhibitory agents.

Qualitative changes in bacterial flora during freezing

Freezing imparts a selective action on the bacterial flora of fish and various bacterial
species are affected at different levels. But Gram-positive bacteria are more resistant to freezing
and frozen storage than Gram negatives. But among Gram positives, differing sensitivity to
freezing is noticed. Bacillus,Lactobacillus and Micrococcusare more susceptible to freezing
than ‘Coryneform’ bacteria. Among the gram negatives, sensitivity to freezing is almost
comparable except the case of Vibrio species that are very much sensitive. Pseudomonas and
Aeromonas, Moraxella and Flavobacterium species are more resistant to the lethal action of
freezing.

Effect of thawing
Generally frozen sea foods are considered highlystable and less prone to bacterial
decomposition. But they can never be considered as sterile and completely free from bacteria. If
there is no contamination after thawing the micro-flora on the thawed seafood will have only the
bacterial population consisting of the freezing resistant species only. This dormant population is
inactive during the frozen storage period. But when the product is defrosted (thawed) surviving
bacteria are liberated and immediately begin multiplying resulting in the chemical breakdown of
the product.

During thawing process, bacteria start multiplying within the temperature limit
characteristic for the different species. The higher the external temperature the more
favorable for outgrowth of most of the bacteria. Hence, defrosting at a lowered temperature
ensures a lower multiplication rate. Multiplication rate of bacteria will be slower at low
temperatures of thawing. As in the case of defrosting temperature, defrosting rate is also found to
have a decisive role in bacterial multiplication and the resulting spoilage. A slow rate of thawing
allows the bacteria to multiply considerably causing the spoilage of the product. Compared to
other foodstuffs, there are greater chances of spoilage of seafood during and after thawing
because fish harbours a psychrophilic bacterial flora even from the beginning. Though freezing
and immediate storage kill as much as 50% of the psychrophilic organisms, sufficient numbers
remain to promote spoilage of thawed frozen product. When frozen fish is defrosted and kept in
the refrigerator, they undergo spoilage similar to that of unfrozen fish.

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Unit 6: Cold stores and containers

● Chapter 1: Construction of cold store

● 6.1 Construction of cold store

A cold store is any building or part of a building used for storage, at temperatures controlled by
refrigeration at - 18ºC or lower.

Recommended Storage Temperature

Reducing the storage temperature can slow all the spoilage of fish due to protein denaturation, fat
changes and dehydration. The FAO Code of Practice for Frozen Fish recommends that frozen
fish products should be stored at temperatures appropriate for the species, type of product and
intended time of storage. The International Institute of Refrigeration recommends a storage
temperature of -18ºC for lean fish such as cod and haddock and -24ºC for fatty species such as
herring and mackerel. The code also recommends that for lean fish intended to be kept in cold
storage for over a year, the storage temperature should be -30ºC.

Practical storage lives (PSL) of fish products

Storage life in months

Product
-18ºC -24ºC -30ºC

Fatty fish (glazed) 5 9 >12

Lean fish (fillet) 9 12 24

Flatfish 10 18 > 24

Shrimp 5 9 12

(cooked/peeled)

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 6.1.1 Shape and size

Cold stores can be divided according to construction into single-storey and multi-storey
buildings. They can be used as production stores, bulk stores, distribution stores or retail stores.
For a long time, the most appropriate shape was a cube for which the ratio of surface to volume
is a minimum. This resulted in multi-storey buildings, with a number of disadvantages, e.g.,
costly foundations, heavy framework, congested handling areas.

The main considerations which have resulted in the appearance and success of single-storey
buildings are cost reduction together with mechanized handling techniques.

A single-storey cold store can be easily designed to meet the specific requirements of stacking
and handling equipment. Wall and roof constructions can be made lighter as they do not have to
support the weight of the product stored, as in a multi-storey building. The main disadvantages
are the relatively large ground area covered and the high ratio of surface to volume. The
advantages however, normally override the disadvantages.

6.1.2 Insulation

The choice of insulation is very important as it accounts for a large proportion of the total
construction cost. The insulation material and thickness is also important from an energy point of
view. Besides a satisfactory thermal conductivity coefficient the insulation material should also
be odour-free, anti-rot, vermin and fire-resistant and impermeable to water vapour.
Typical insulation thickness for chill and cold stores utilizing different insulation materials

Calculated thermal conductivity

Insulation Thickness (mm)
(kcal/m hoC

Polystyrene 0.033 220

Styrofoam FR 0.030 200

Polyurethane 0.025 170

Vapour Barriers
Water vapour in the air gives rise to a pressure and together with the other gases present. The
partial pressure exerted by the water vapour is proportional to the quantity of vapour present and
the vapour in the air will tend to migrate from areas of high partial pressure to areas of low
partial pressure. Hence, there is a tendency for moisture in the ambient air to pass through the
insulation of a cold store to the area of low partial pressure within. When this vapour is cooled, it

90
condenses and the build-up of ice will eventually affect the insulation properties of the cold store
wall and also weaken the structure of the wall or building.
Foundations and Frost Heave
Low temperature stores built directly on the ground may require special precautions to prevent
the build-up of ice below the cold store floor. The ice formation causes distortion known as
“frost heave” (fig.) and in particularly bad cases, it can lead to the complete destruction of the
structure of the building. The conditions that give rise to frost heave are rather complex, since
they are related to the type and texture of the soil, the insulation properties, the availability of
moisture, the dimensions of the store, seasonal climatic variations and other factors. Two
methods of preventing frost heave are commonly used. The ground below the store can be heated
either by a low voltage electrical mat in the cold store foundation or by circulating a heated
liquid such as glycol through a pipe grid built into the foundation. The heat for the glycol is
usually obtained from the compressor hot gas through a heat exchanger.

6.1.3 Air ingress

Outside air entering the store adds heat and moisture. This moisture will be deposited as frost on
any cold surface and will eventually cover the surface of the cooler. Excessive air exchange
should be prevented to keep the cold store temperature steady and reduce the frequency of
defrosting. Small air locks have been used to prevent the free flow of air in and out of the store.

A curtain of air blown downward or from the side of the doorway can reduce the exchange of air
when the door is open. These air curtains, as they are called, can be a useful aid when the door is
opened for short intervals. However, they are often abused and doors are often left open for long
periods.

Floors

The ground loads from a cold store are in the order of 5500-8000 kg/m2. This consists of static
loads due to merchandise, structure and concentrated rolling loads transmitted by e.g., forklift
trucks and other handling equipment. It is of importance that those loads are investigated in
detail for each special project. Most commonly the floor wearing surface is a concrete slab cast
on the floor insulation with a thickness of 100-150mm. In cases where intensive traffic is
foreseen a special hardwearing top-finish is recommended. Before casting the wearing surface,
the floor insulation should be protected by bituminous paper or plastic sheeting, the function of
which is twofold.

6.1.4 General layout

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A single-storey building can either have one single room or it can be divided into a number of
rooms. Normally all the rooms are operated at the same temperature, for fish preferably in the
range of –24o to -30oC. Most stores are built at a higher level than the surrounding yard with a
special loading ramp at one or more sides. The loading ramp level corresponds to the height of
the most commonly used vehicles.

The engine room should be as close as possible to the position of the air cooling equipment
within the store.

Modern large or medium cold stores are built as one-storey buildings designed for mechanical
handling, e.g., forklift trucks and automatic stacker cranes. Manual handling is, however, still
used for most small-sized stores.

A cold store can be built as an ordinary building using conventional building material, such as
bricks, concrete or concrete precast sections to which a vapour barrier and insulation is fitted
internally. Modern insulation material, in particular polyurethane, has a strength that can be
utilized structurally. Today, this is used for panel designs suitable for all sizes of cold rooms from
20m3 to 250 000m3. Factory made insulation panels are delivered to the site complete with a
vapour barrier and internal cladding, thus reducing the site work to a minimum. There are two
basic principles for panel-built cold stores. A common system has an external structure and
cladding with wall insulation inside of the columns and the insulated ceiling hanging from the
outer roof structure.

The panels normally used in these systems are either polyurethane or polystyrene insulated
panels with or without frames. They are manufactured as sandwich panels, one face being the
vapour barrier of light-gauge galvanized steel sheet and the other face being the internal finish of
plastic-coated galvanized sheet or aluminum sheet. A decorative external cladding is erected on
the outside of the columns. The roof insulation is constructed as a suspended ceiling. The roof
panels are, in principle, the same as the wall panels, but are sometimes equipped with wooden
frames.

The latest development are panels with a rib profile on the external face of aluminum – with
polyurethane foam insulation and an internal face of low profile corrugated aluminum. The space
between the panels is filled with one component polyurethane foam. The internal cladding is
sealed with a PVC strip. One component foam is also used to join wall panels to roof panels and
seal around doors, etc, maintaining good insulation properties throughout the building. With this
design both the insulation and external vapour barrier are entirely sealed units enveloping the
whole building. This means that losses via heat bridges or air leakage are completely eliminated,
which gives practical insulation properties closer to theoretical values than normally expected.

Chapter 2: Refrigerated containers

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 6.2.1 Refrigerated Containers

In a refrigerated transport container or insulated vehicle it is difficult to maintain a low and stable
produce temperature. Added to this, the effect of fans, the opening of the door and the presence
of workmen is proportionately greater in refrigerated transport than in a large, well-designed
storage chamber. Under operating conditions the best temperature obtainable is approximately
-180C to -150C.

It follows that produce should be kept in transport vehicles and containers for as short a time as
possible. Very often parked containers are used for cold storage to avoid the expense of
constructing a cold store. However one has to keep in mind that parked transport containers and
vehicles are not suitable for use as stores for frozen produce except for very short periods and in
emergencies. There is such greater risk of loss of quality through temperature fluctuation and
desiccation in refrigerated containers than in large, well-designed and well-operated cold store.
Storage in transport containers is also very expensive when compared with properly designed
cold stores.

The Cold Chain

Temperature is the single most important factor influencing fish spoilage. The cold chain is a
temperature chain. It means 00C for fresh fish and -200C for frozen fish from the producer to the
consumer. Temperature rise and temperature fluctuation will accelerate quality deterioration and
it should to be avoided. Especially sensitive to temperature abuse are fresh, live and lightly cured
(e.g. smoked fish) products. These should be kept at 0-10C, and their temperature should never
rise above 30C.

Temperature fluctuations accelerate the reformation of small ice crystals into larger crystals
damaging the cells. Also, desiccation will increase because of temperature fluctuations.

The insulated transport vehicles and containers shall be provided with adequate quantities of
coolant or good mechanical refrigeration. Refrigeration requirements vary considerably
depending on the duration of transportation, the temperature at which a product has to be
maintained, the quality of insulation material, the dimensions of a container and the temperature
of the environment.

6.2.2 Loading

The transport vehicle has to be pre-cooled for several hours prior to loading. With pre-cooling a
large rise of product temperature will be avoided. Furthermore frozen and chilled fish should not
be left outside for a considerable length of time and exposed to high temperatures. To avoid such
exposure fast loading and unloading is required.

Transit Points

93
This shipper should take into consideration that transit points like airports often do not have
proper refrigeration facilities, or because of mismanagement, chilled consignments are not stored
in refrigerated rooms while in transit. When no refrigeration facilities are available, more coolant
should be used. Alternatively the consignment might be re-iced during transit. The best way to
avoid temperature abuse at transit points is to have good organization and planning.

Temperature Monitor

To check on temperature abuse, continuously working monitors have been developed. These give
proof of proper temperature control or abuse during road transport, at airport terminals and so on.
Because of the financial risks involved it is necessary to use such monitors especially with large
or high value consignments.

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Chapter 3: Good handling andshipping practices

6.3.1 Fresh fish

The important factors in the distribution of fresh fish are speed, cleanliness and the need to keep
the fish well chilled. The fish must not be handled repeatedly.
Everything likely to come into contact with the fish has to be clean. Bacteria may be picked up
from dirty surfaces, tools, uniforms and hands. This will accelerate, fish spoilage and may
endanger public health. Fish boxes, trucks, loading platforms have to be cleaned and disinfected
before use.
For shipping fresh fish, refrigerated transport is required. Temperature is the single most
important factor influencing fresh fish spoilage. Chill fresh fish with ice immediately and keep it
cool. Mix the fish and ice well. Ice must always remain in sufficient quantities for effective
chilling and to separate the fish from each other and from the supporting surfaces. The air
temperature in the container should be 2-30C to allow the ice to melt slowly.
Fresh Fish Spoilage

● Coldwater fish spoils faster than warm water fish.

● Fatty fish spoils faster than lean fish.
● Smaller sized fish spoil faster than larger fish of the same species.
● Round fish spoils faster than flat fish.
● Shark spoils faster than lean fish.

Fresh Fish Fillets

Direct contact between fillets and ice is not recommended. Water will soften the texture and
affect appearance. Separate the ice from the fillets e.g. by using plastic sheeting. Be sure to chill
the fillets properly before packing. Chilling after packing is difficult because of the tightly
packed mass of fish flesh.
Frozen Fish

For shipping frozen fishery products refrigerated transport is required. The cold chain has
to be maintained at all times. The normal working temperatures in modern cold stores for quick
frozen foodstuffs are -30 to -250C. Retail cabinets, in which the product should be stored for no
more than a few weeks, usually operate at about -180C or a little lower.

Do not try to freeze fresh produce or try to lower the temperature of frozen fish when
shipping. Prior to shipment always check the product temperature with a thermometer. Frozen
fish should be handled with care. Rough handling, such as throwing, will cause the frozen flesh
to crack, damage the skin and fins and these defects will inevitably show up when thawed.

Unit 7: Packaging methods

95
Chapter 1: Packaging for fresh fish handling

7.1.1 Packaging for Fresh Fish Handling

To maintain the quality of fresh fish, use of good fish boxes and packaging materials is required.
Specially designed and constructed fish boxes are now widely used, such as insulated containers
for air shipment. Appropriate packaging also makes handling easier, which reduces labour costs.

Important Requirements for Fish Boxes and Containers

● They must be of a suitable size for the range of fish to be handled or the products to be
put into them.
● They should be of a convenient size for manual handling or lifting by mechanical
equipment.
● They should be stackable such that the weight of the containers on top rests on the
containers underneath and not on the fish.
● They should be constructed of impervious non-tainting materials.
● They should be easy to clean.
● They should provide drainage for the melt –water.

Polyethylene Fish Boxes

High density polyethylene is most commonly used for fish boxes as it is strong and light in
weight and easy to clean. Polyethylene fish boxes are very popular onboard fishing vessels.

Disposable Fish Boxes

The disposable boxes are commonly used. These hold up to approximately 25 kg of fish or fish
plus ice. They may or may not be insulated. Insulated boxes include fiberboard boxes with
polystyrene liners.Non-insulated fish boxes are normally fiber board cartons, waxed or otherwise
water-proofed. A road transport, disposable packaging can be a water-proofed carton with or
without insulation depending on whether the transport vehicle is refrigerated or not. Packaging
for air transport, has to be light, strong and leak proof, may consist of a heavy duty
water-proofed corrugated carton with two polyethylene liners and insulating material between
the liners.

7.1.1.1 Moulded polystyrene Fish Box

Moulded polystyrene fish boxes are commonly used for delivery of chilled fish and frozen fish.
In a typical range the wall thickness varies with the box size, for example a 6 kg capacity box has
a 15 mm thick wall, a 10 kg box a 19 m wall, a 25 kg box a 25 mm wall. The major
disadvantages of polystyrene boxes are their lack of strength. This limits their size and use. They

96
are easily damaged or broken by rough handling. Polystyrene is difficult to clean. Polystyrene
boxes are difficult to re-use, and are normally non-returnable .

Use of Insulating Linings

Wooden or fiber board boxes can be insulated with polystyrene sheeting cut to the required
dimensions. Alternatively waxed fiber board which has good water resistance can be used. The
empty box can be shipped in a knocked down form. This is cheaper than using moulded
polystyrene boxes.

Light-Weight Insulated Packaging

A recent development in light-weight insulated packaging makes use of the ability of metallised
surfaces to reflect practically all (97%) radiant heat and so to reduce the thermal insulation
thickness. The thermal guard packaging system consists of a metallised plastic bag in which the
produce is packed, a bubble-pack insulating wrapper, provides the insulation. The thermal guard
bag can be sealed airtight and can thus be used for frozen fish.

Returnable Versus Disposable Containers

The environmental pollution, authorities in many countries try to reduce the use of disposable
containers. The returnable containers are the freight costs for returning empty containers, the
logistics involved and the high investment in good quality containers. Using a ‘knock –down’
returnable container will reduce return freight costs. The returnable containers are to be preferred
because in the long run these will then be cheaper and generate less trash.

7.1.1.2 Refrigerants and Coolants

Ice should be properly mixed with the product and applied in sufficient quantities to last for the
duration of the shipment. When fish are in close contact with each other or with a smooth
impervious surface such as the inside of a box, anaerobic bacteria producing offensive odors may
develop. The air temperature in the container should be 2-3ºC to allow the ice to melt slowly.
Melt-water keeps the fish well chilled and moist. The melt-water also helps to wash away the
products of spoilage. The melt-water should be drained away from the produce.

Alternative refrigerants to wet ice are dry ice and gel ice. Dry ice is solid carbon dioxide.
Evaporation of the dry ice gives the cooling effect. Because of its low temperature (-78.9ºC), dry
ice should not be in direct contact with chilled fish, to avoid cold burns.

Gel ice is made by freezing a water based gel. The advantage of gel ice is that all the water is
bound with no danger of water leakage during thawing. It is often claimed that gel packs release
the ‘cold’ more slowly and equally than other coolants so that they are superior in maintaining
chilled temperatures.

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Chilling air cargo

With air cargo often preference is given to dry ice or gel packs, as these do not cause
leakage. However the use of dry ice is subject to restrictions because it expands from a solid
form into a gas and it may ‘drive-out’ oxygen. This obviously poses a hazard to the safety of
airplanes. The international identification number for dry ice is UN 1845.

7.1.1.3 Guidelines for air shipment of sea food

● Boxes, cartons or other outer packaging materials must be multiwall, was impregnated or
waterproof coated on both inside and outside. Gusseted corners must be on both top and
bottom with a top that extends fully over to the bottom.
● Outer packaging must be banded in two directions.
● Use dry-ice or gel ice preferably.
● Product must be sealed in a polyethylene bag with wet ice a minimum 4 mm gauge
polyethylene bag or liner is required.
● Use an absorbant pad.
● Styrofoam boxes are to be protected by wet lock cartons.
● The cartons must clearly indicate the airway bill number, name of the shipper,
consignee’s name and address, plus marking like “This Side Up” and “Perishable Fresh
Sea foods”.
● With dry ice follow the IATA Dangerous Goods Regulations to declare use and amount of
dry ice.

Chapter 2: Packaging of frozen fishery products

7.2.1 Introduction

The factors that must be taken into account while selecting a package for frozen foods are,
protection of the contents from

● atmosphere oxygen
● loss of moisture
● flavor contamination
● entry of microorganisms
● mechanical damage
● exposure to light

In addition to the above, the materials used must have a high heat transfer rate to facilitate
rapid freezing. The package shall have an aesthetic appeal and promotes sales of product and
hence a well designed package is sometimes called the ‘silent salesman’. Quick freezing food
require transparent, moisture vapour proof, wet strength, tasteless and odourless wrapping
materials.

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 Modern frozen food processors can select from a wide range of protective materials such
as variety of papers (Kraft papers coated with wax or plastic and laminated papers), films
(moisture proof, heat sealable cellophane and various thermoplastic films such as saran, mylar,
pliofilm and plastic wrap) and foils (heavy aluminum foil) in designing the package.
The films are transparent, lightweight, resistant to puncture and flexible at low temperatures, free
from off odours and off flavours, non-toxic, heat sealable and have a low moisture vapour
transmission rate.

7.2.1.1 Primary packages

They are three forms a plastic packaging:

1. Flexible films,
2. Semi rigid packages,
3. Rigid packages,

It is estimated that more than 600 flexible film combinations are commercially available today.
Terms frequently used in describing flexible film packages are coatings and laminations.

7.2.1.2 Flexible films

1. Flexible films

A variety of flexible films is used in frozen food packaging as bags, pouches and over
wraps. Almost all of these films are made from organic polymers. Some of the natural polymers
such as starch, cellulose and rubbers are also useful in food packaging.

Polyethylene (PE): Polyethylene is a long chain polymer of ethylene, a gas derived from
natural gas or petroleum. It is an useful film in frozen food packaging. It is low in cost, has
excellent clarity and gloss, is resistant to tearing, has relatively low WVTR, is flexible at low
temperatures and is heat sealable. Because of the last property, it is frequently used as coating or
as layer in a laminate with heat sealable films. Important properties such as heat seal ability,
moisture resistance and flexibility depend on the density of the polyethylene resin from which it
is made. Polyethylene is classified into three density ranges and expressed as g/cc as follows.
(1) Low – 0.910 to 0.925
(2) Medium – 0.926 to 0.940
(3) High – 0.941 to 0.965
Most polyethylene films used in frozen food packaging are in the low and medium density
ranges.
Nylon (Polyamide PA):It is a group of long chain polymers with re occurring amide groups as a
part of the chain. Nylon films have excellent properties of toughness, tear and breaking strength.
They are useful both as single films and in many laminates.

Plio films:It is the trade name for rubber hydrochloride film. It was the first transparent film
which could be heat sealed.

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Polypropylene (PP):It is relatively new film in frozen food packaging, but its clarity, strength at
low temperatures and low WVTR make it useful in laminates, and in coating paper or aluminum
foil. It is heat sealable, when it is heated with a variety of coatings.

Cellophanes:Cellophane is made from regenerated cellulose. When it is coated with other

plastic resins such as polyethylene or saran, a wide variety of cellophanes result, each designed
for a specific use. These make excellent packaging films. They are strong, moisture proof, heat
sealable and flexible at low temperatures.

Poly vinylidene chloride (PVDC):Polymers of vinylidene chloride are better known as saran
and are available not only as films, but as water based emulsions in which form they are
frequently used as coating resins for paper substrates. Saran is superior to polyethylene as a
water vapour barrier material and has low oxygen permeability. The film is not heat sealable.
The film is excellent for use as a wrap around odd shaped meat (shrink wrapping).

Polyesters (PES):This strong film is well known under the trade name of Mylar. It is frequently
used in lamination with other films, especially withpolyethylene to make pouches which can be
filled and heat sealed. These laminates are now used for boil in the bag pouches. Polyester films
are strong, durable, transparent, non-toxic, inert and easily stretched. They also have the
advantage of being flexible at very low temperatures (-710C). They are sometimes known as
nylon type films and because of their roughness,they are useful in packaging sharp pointed,
irregular foods such as lobsters.

2. Plastic Pouches
The pouch is usually made from a laminate of polyethylene, nylon and Mylar films. One
of the weakest links in plastic bags is the seal, both from the stand point of vapor leakage and
that of bag breakage at the seal.

7.2.1.3 Shipping container

Individual packages must be further packed in a properly engineered container for shipping and
storage. Corrugated containers are the most widely used form of containers for shipping frozen
foods. Wooden crates and foam polystyrene shipping containers are also used. The use of a
corrugated sheet provides exceptional crush resistance. It also has a high capacity for absorbing
shock.

Corrugated fiber board is made of fluted (grooved) sheet glued between two liners. Four
types of corrugated board are commercially used and they are A,B,C and E flute. The difference
lies in the number of flutes and board thickness. ‘A’ flute -35 flutes per inch and 3/16 inch
thickness; ‘B’ flute -50 flutes per inch and 1/8 inch thickness; ‘C’ flute -41 flutes per inch and
5/32 thickness; ‘E’ flute -90 flutes per inch and 5/64 inch thickness. Each type has some
specifications and advantage over the others.

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 Though various coating methods have been used, the most recent technique is called
‘curtain coating’, wherein the coating formulation is applied by a machine designed for that
purpose. The continuous film coating acts as an effective barrier, not only against free water, but
also against water vapor, gas, grease, oil, and other foreign substances. It is necessary to use a
coated shipping container, if there is a possibility of the container being exposed to high
humidity or excessive moisture conditions during handling.

The foam polystyrene container for shipping fresh frozen foods is relatively new. It has the
advantage of being water resistant and having excellent insulating properties. After the frozen
product is placed inside, the foam lid is applied, and the container sealed off. It has the
disadvantage of being somewhat more costly and hardens to handle. The empty containers are
bulky to handle and store. However, because of its light weight and insulating properties, the
polystyrene container is particularly adaptable to the air transit of frozen foods.

7.2.1.4 Shrink packaging

This type of packaging was introduced by the Cryovac company in 1948 as a protective
package for frozen poultry. The saran-type film which was first used for this kind of packaging
offered excellent protection against freezer burn, and loss of weight by desiccation. In the
cryovac process, the whole bird, or a large piece of red meat is inserted into a plastic bag, air is
removed by vacuum so that the bag is drawn in tightly against the bird, the bag is sealed by
twisting and clamping and film is shrunk into a skin-tight wrap by dipping the package into hot
water (about 900C) for a few minutes.

7.2.1.5 Requirements for packaging materials

The requirements for packaging materials for frozen foods to be measured include

1. tensile strength and elongation

2. tear strength
3. impact strength
4. stiffness
5. bursting strength
6. fold endurance
7. compression
8. grease resistance
9. water vapor transmission.

A packaging material may be water proof without being water vapor proof. A water proof
material will hold water and will not go to pieces in water. A water-vapor proof material prevents
vapor from passing through it.

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Packaging Machinery
The packaging operation can be divided into four major steps

1. Forming the package

2. Filling the package
3. Closing and sealing the package
4. Placement in the shipping container

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Chapter 3: Vaccum packaging and modified atmosphere packaging and storage

7.3.1 Vaccum packaging

● It is a viable technology in seafood processing.

● It is wide spread in the meat industry for fresh, processed and cured products but has
faced difficulties in its application to seafood.

Condition in the vaccum package

It is brings several changes

1. A drastic reduction in O 2 content

2. An increase in CO2
3. Increase in acidity

Advantages:

● It increase the shelf life of fish (ie., It is very good barrier to O2)
● Variety of spoilage organisms is substantially decreased
● Microbial growth is prevented entirely (Partially)

Packaging Materials

Light, water, heat, O2, bacteria and are consider to be an affecting factors in this packaging.

Light

The length of the exposure, the wave length and intensity of light and permeability of the
package to the light affect nutrients.

Temperature

This type of packaging material greatly affect the rate at which heat i.e. transferred to and from
the food.

This is important for both refrigerated and frozen seafood.

Insulating containers like Styrofoam food trays greatly reduce the transfer of heat of the fish.

Oxygen

Excessive amount of O2 promote seafood deterioration through the oxidation of fats, the
degradation of some amino acids and the destruction of certain vitamins.

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The effects of O2 are related to the atmosphere pressure,humidity and temperature.

Exposure of the package contents to O2 may occur through loose closure, puncture and or
diffusion through the packaging materials itself. Permeability of O2 also affected by the moisture
and temperature.

Disadvantages of vaccum packing

 Doesn’t ensure free from microbial activity.

 The O2 impermeable materials required are relatively expensive and appropriate
equipment is needed.

7.3.2 Modified atmospheric packaging and storage

● In 1932, conye reported the beneficial effects of CO2 in inhibiting the growth of bacteria
from fish.
● They have also been applied to other storage systems, namely CO2 saturated sea water
and CO2 enriched atmospheres.
● Increased storage life of fresh fish and shell fish using refrigerated sea water saturated
with CO2 become evident.
● The results appear to apply to a wide variety of species including fatty species of salmon,
low fat rock fish and shrimp.
● When the air surrounding the fish is changed in composition usually by enriching it with
CO2 and the container then sealed the atmosphere is called “modified”.
● Fish and shell fish from modified atmosphere storage have lower bacterial counts, greater
acidity level, reduced trimethylamine formation and lower NH3 production.
● There is also residual and positive effect on the keeping quality after removal from the
CO2 enriched environment.
● Temperature abuse results in bacterial spoilage and possible development of toxins. The
accumulation of unpleascent odour in the package owing to the activity of anaerobic
organisms.
● The more serious problem is the threat of C. botulinum activity
● The odour are not the usual type of “fishy” odours produced by sea food because of the
circulation of fresh gas removes them.

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Unit 8: HACCP

● Chapter 1: Hazard analysis and critical control point- A detailed approach

8.1.1 Introduction to the codex Alimentarius commission

The Codex Alimentarius Commission (CAC) was established by FAO in 1961. CAC is
an intergovernmental body with 158 member Governments as on 31st August, 1997. The Codex
Alimentarius (‘Food Code’ or ‘Food Law’ in Latin) is a collection of food standards, codes of
practice and other recommendations presented in a uniform way. Codex standards, guidelines
and other recommendations ensure that food products are not harmful to the consumer and can
be traded safely between countries. The Codex Alimentarius has resulted in evaluation of the
safety of over 760 food additives and contaminants and setting of more than 2500 maximum
limits for pesticide residues and more than 150 for veterinary drug residues.

CAC has adopted the guidelines for the application of the Hazard Analysis Critical
Control Point (HACCP) system through its committee on Food Hygiene. It has recognized
HACCP as a tool to assess hazards and establish control systems that focus on preventive
measures instead of relying primarily on end product testing.

The Uruguay Round of Multi lateral Trade Negotiations which concluded in 1994 established
the World Trade Organization (WTO) to replace the General Agreement on Tariffs and Trade
(GATT).

1. The Agreement on the Application of Sanitary and Phyto Sanitary Measures (SPS
Agreement) The SPS Agreement confirm the right of WTO member countries to apply
measures necessary to protect human, animal and plant life and health.
2. The Agreement on Technical Barriers to Trade (TBT Agreement). The TBT Agreement
basically provides that all technical standards and regulations.

8.1.2 Work of the codex alimentarius commission

The work of the codex Alimentarius Commission – including the guidelines for the
application of HACCP system – has become reference for international food safety requirements.
The application of the General Principles of Food Hygiene and Good Manufacturing Practices
(GMPs) allows the producer to operate with in environmental conditions favourable to the
products.

The main objectives of the Codex General Principles of Food Hygiene are

1. Identify the essential principles of food hygiene, applicable throughout the food chain to
achieve the goal of ensuring that food is safe and suitable for human consumption
2. Recommended an HACCP based approach as a means to enhance food safety
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 3. Indicate how to implement those principles and
4. Provide guidance for specific codes which may be needed for sectors of the food chain,
processor, or commodities and to amplify the hygiene requirements specific to those
areas.

5. 8.1.3 History of HACCP

Dr.Deming and others developed total quality management (TQM) systems for Japanese
products in the 1950’s.NASA wanted a “Zero defects” programme to guarantee the safety of the
foods that astronauts would consume in space. Pillsbury presented the HACCP concept publicly
at a conference for food protection in 1971. The use of HACCP principles in the promulgation
of regulations for low-acid canned food was completed in 1974 by the United States Food and
Drug Administration (FDA). In the early 1980’s the HACCP approach was adopted by other
major food companies.

The International Commission on Microbiological Specifications for Foods (ICMSF) and

the International Association of Milk, Food and Environmental Sanitaries (IAMFES), have
recommended the broad application of HACCP to food safety.

ADVANTAGES OF HACCP

The HACCP system, as it applies to food safety management, uses the approach of
controlling critical points in food handling to prevent food safety problems. The system, which
is science based and systematic, identifies specific hazards and measures for their control to
ensure the safety of food. The application of the HACCP system can aid inspection by food
control regulatory authorities and promote international trade by increasing buyers’ confidence.

Guidelines for the application of the HACCP system

Management commitment is necessary for implementation of an effective HACCP

system. During hazard identification, evaluation and subsequent operations in designing and
applying HACCP systems, consideration must be given to the impact of raw materials,
ingredients, food manufacturing practices, role of manufacturing process to control hazards,
likely end use of product, categories of consumers of concern, and epidemiological evidence
relative to food safety.

The intent of the HACCP system is to focus control at CCPs by all means.

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 8.1.4 Principles of the HACCP system

1. Conduct a hazard analysis

2. Determine the critical control points (CCPs)

3. Establish critical limit(s)

4. Establish a system to monitor control of the CCP

5. Establish the corrective action to be taken when monitoring indicates that a

particular CCP is not under control

6. Establish procedure for verification to confirm that the HACCP system is working
effectively

7. Establish documentation concerning all procedures and records appropriate to these

principles and their applications

Logic sequence of application of HACCP

Assemble HACCP

Describe product

Identify intended use

Construct flow diagram

On-site confirmation of flow diagram

List all potential hazards conduct a hazard analysis, consider control measure

Determine CCPs
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 

Establish critical limits for each CCP

Establish a monitoring system for each CCP

Establish corrective actions

Establish verification procedures

Establish documentation and record keeping

8.1.5 The HACCP team

Team composition:

When selecting the team, the coordinator should focus on:

o Person who will be involved in hazard identification

o Person who will be involved in determination of critical control points
o Person who will monitor CCPs
o Person who will verify operations at critical control points
o Person who will examine samples and perform verifications procedures

Knowledge required:

Selected personnel should have a basic understanding of:

o Technology and equipment used on the processing lines

o Practical aspects of the food operations,the flow and technology of the process
o Applied aspects of food microbiology and HACCP principles and techniques

Product description:

1. Product name (common name) or group of product names (the grouping of like products
is acceptable as long as all hazards are addressed)

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 2. Important end-product characteristics: properties or characteristics of food under review
those are required to ensure its safety (e.g. aw, pH, preservatives)
3. How the product is to be used (i.e. ready to-eat, further processing required, heated prior
to consumption)
4. Type of package, including packaging material and packaging conditions (e.g. Modified
atmosphere)
5. Shelf-life, including storage temperature and humidity if applicable
6. Where the product will be sold ( e.g. Retail, institutions, further processing)
7. Labelling instructions (e.g. Handling & usage instructions)
8. Special distribution control ( e.g. Shipping conditions)

8.1.6 Form. 2 Product ingredients and incoming material

List the product ingredients and incoming materials (including raw material, product ingredients,
processing aids, and packaging materials) that are used during the manufacturing process. This
exhaustive listing is required for proper identification of all potential hazards that could apply.

Flow diagram: - (will identify the important process steps)

o All ingredients and packaging used (biological, chemical, physical data)

o Sequence of all process operations (including raw material addition)
o Time/ temperature history of all raw materials and intermediate and final
products, including the potential for delay.
o Flow conditions for liquids and solids
o Product cycle/rework loops
o Equipment design features

Plant schematic:

This plan should aid in the identification of any areas of potential cross-contamination with in
the establishment. The plant schematic/floor and equipment layout should be considered in detail
and assessed. Data may include but is not restricted to:

o Personnel routes
o Routes of potential cross – contamination
o Area segregation
o Flow of ingredients and packaging materials
o Location of changing rooms, wash rooms, lunch rooms and hand-washing stations

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 8.1.7 Hazard analysis: (Principle 1)

Hazard will vary among firms making the same products because of differences in:

● Source of ingredients
● Formulations
● Processing equipments
● Processing and preparation methods
● Duration of process
● Storage conditions
● The experience, knowledge and attitude of personnel

Therefore hazard analysis must be done on all existing and new products. Changes in raw
materials, product formulations, processing or preparation procedures, packaging, distribution
and/or use of the product will require review of the original hazard analysis.

Examples of biological hazards: Viruses

Bacteria (Spore forming): Hepatitis A & E

Clostridium botulinum Norwalk virus group

C.perfringens Rotavirus

Bacillus cereus Protozoa & parasites:

Bacteria (Non-spore forming): Diphyllobolthriumlatum

Brucella abortis Entamoeba histolytica

B.suis Giardia lamblia

Campylobacter spp Ascaris lumbricoides

Cryptosporidium parvum Taenia solium

(E.coli 0157; H7, EHEC,EIEC, Taenia saginata

TEC, EPEC) Trichinella spiralis

Listeria monocytogenes

Salmonella spp.

S.typhimurium S. enteriditis

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 Shigella spp .

Staphylococcus aureus

S. pyogenes

Vibrio cholerae

V.Parahaemolyticus

V.vulnificus

Yersinia enterocolitica

8.1.8 How to conduct a hazard analysis

After listing all the hazards (biological, chemical or physical) that may be reasonably
expected at each step from primary production, processing, manufacturing and distribution until
the point of consumption, the HACCP team should assess the potential significance or risk of
each hazard by considering its livelihood occurrence and severity. The estimate of the risk of a
hazard occurring is based upon a combination of experience, epidemiological data and
information in the technical literature. Severity is the degree of seriousness of the
consequences of a hazard if the hazard is not controlled.

Hazard addressed under the HACCP system must be of such a nature that their
prevention, elimination or reduction to acceptable levels is essential to the production of safe
foods. Hazards of a low probability of occurrence and a low severity should be addressed under
the HACCP system but may be addressed through the good manufacturing practices (GMPs)
contained in the codex general principles of Food Hygiene.

8.1.9 Hazard assessment

Severity:

Severity is the magnitude of a hazard or the degree of consequences that can result when a
hazard exists. Disease causing hazards can be categorized according to their severity. One system
uses the categories of:

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 ● High (life-threatening ) ex. include illness caused by C.botulinum, S.typhi, Listeria
monocytogenes, E. coli 0157 H7, V.cholerae, V. vulnificus, paralyticshell fish
poisoning and amnesic shell fish poisoning.
● Moderate (severe or chronic) eg. include illness caused by Brucella sp. Campylobacter
sp. Salmonella sp, Shigella sp, Streptococcus sp, Yersinia entrocolitica, hepatitis A virus,
mycotoxins, Ciguatera toxin
● Low (moderate or mild) – ex. include illness caused by Bacillus sp. C.perfringens,
Staphylococcus aureus. Norwalk virus, most parasites, histamine – like substances and
most heavy metals that cause mild acute illnesses.

Risk is a function of the probability of an adverse effect and the magnitude of that effect,
consequential to a hazard(s) in food. Degrees of risk can be categorized as high (H), Moderate
(M) low (L) & negligible (N).

8.1.10 Critical control points: Determination (Principle 2)

The determination of CCPs is the 2nd principle of HACCP. The codex guidelines define a
CCP as a step at which control can be applied and is essential to prevent or eliminate a food
safety hazard or reduce it to an acceptable level. The determination of a CCP in the HACCP
system can be facilitated by the application of a decision tree such as that included in the codex
HACCP system and guidelines for its application which indicates a logical reasoning approach.

Fig. Example of decision tree to identify critical control points.

Q1. Do control preventive measures exist? 

 

Yes No

 

 Is control at this step  Yes  Modify step, process

or

products

¯ Necessary for safety?

 

 No  not a CCP  Stop *

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Q2. Is the step specifically designed to eliminate or reduce the likely occurrence of hazards to an
acceptable level?

 

No Yes  CCP.

Q3. Could contamination with identified hazards occur in excess of acceptable levels (or) could
this increase to unacceptable levels? **

 

Yes No  Not a CCP  Stop *

Q4. Will a subsequent step identified hazard(s) or reduce likely occurrence to an acceptable level
?

 

Yes No  CCP

Not a CCP  Stop *

*Proceed to the next identified hazard in the described process

** Acceptable and unacceptable levels need to be defined within the overall objectives in
identifying the CCPs of the HACCP plan.

8.1.11 Establish critical limits for each CCP (Principle 3)

Critical limits:

Critical limits are defined as criteria that separate acceptability form unacceptability. A critical
limit represents the boundaries that are used to judge whether an operation is producing safe
products. Critical limits may be set for factors such as temperature, time, physical product
dimensions, water activity, moisture level, etc. These parameters, if maintained within
boundaries, will confirm the safety of the product.

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Sources of information on critical limits include

● Scientific publications/Research data

● Regulatory requirements and guidelines
● Experts (eg. Thermal process authorities, consultants, food scientists, microbiologists,
equipment manufactures, sanitarians, academicians)
● Experimental studies (eg. In house experiments, contract laboratory studies)

Table Examples of critical limits

Hazard CCP Critical limit

Bacterial pathogens Pasteurization 72oC for at least 15 sec.

(Non-sporulating)

Metal fragments Metal detector Metal fragments larger than 0.5mm

Bacterial pathogens Drying over Aw< 0.85 for controlling growth in

dried food products

Excessive nitrite Curing room/brining Max. 200 ppm sodium nitrite in

finished product

Bacterial pathogens Acidification step Max. pH of 4.6 to control C. botulinum

in acidified food

Food allergens Labelling Label that is legible and contains a

listing of correct ingredients

Histamine Receiving Max. of 25 ppm histamine levels in

evaluation of tuna for histamine

Table Critical limits Vs Operating limits

Process Critical limit Operating limit

Acidification pH 4.6 pH 4.3

Drying 0.84 A w 0.80 A w

Hot fill 80oc 85oc

Slicing 2.0cm 2.5cm

8.1.12 Establish a monitoring system for each CCP (Principle 4)

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What will be monitored?
Monitoring may mean measuring characteristics of the product or of the process to
determine compliance with a critical limit.
Examples are:

● Measurement of the time and temperature of a thermal process

● Measurement of cold-storage temperature
● Measurement of pH
● Measurement of A w

Monitoring may also mean observing whether a control measure at a CCP is being implemented.
Eg. i) Visual examination of sealed cans, and ii) Verification of vendor’s certification of analysis
Establish corrective Actions (Principle 5)
The codex HACCP system and guidelines for its application defines corrective action as “any
action to be taken when the results of monitoring at the CCP indicate a loss of control”
Loss of control is considered as a deviation from a critical limit for a CCP. Deviation
procedures are predetermined and documented. All deviations must be controlled by taking
actions to control the non-compliant product and to correct the cause of non-compliance. Product
control includes proper identification, control and disposition of the affected product. The control
and disposition of the affected product and corrective actions taken must be recorded and filed.

8.1.13 Establish corrective actions (Principle 5)

The codex HACCP system and guidelines for its application defines corrective action as
“any action to be taken when the results of monitoring at the CCP indicate a loss of control”

Loss of control is considered as a deviation from a critical limit for a CCP. Deviation
procedures are predetermined and documented. All deviations must be controlled by taking
actions to control the non-compliant product and to correct the cause of non-compliance. Product
control includes proper identification, control and disposition of the affected product. The control
and disposition of the affected product and corrective actions taken must be recorded and filed.

8.1.14 Establish verification procedures (Principle 6)

Verification:

Verification is embodied in HACCP principle 6. The codex guidelines define verification as “the
application of methods, procedures, tests, and other evaluations, in addition to monitoring to
determine compliance with the HACCP plan”, Verification and auditing methods, procedures

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and tests, including random sampling and analysis, can be used to determine if the HACCP
system is working correctly.

8.1.15 Establish Documentation and Recorded keeping (Principle 7)

Four types of records should be kept as part of the HACCP programme:

● Support documentation for developing the haccp plan

● Records generated by the HACCP system
● Documentation of methods and procedures used
● Records of employee training programme

Chapter 2: Physical, chemical and biological hazards in seafood

8.2.1 Introduction

Hazard analysis is the assessment of all hazards that are associated with the operational steps
of the process. Three hazards under the food safety are

● Physical
● Chemical and
● Biological

Physical hazards: Bones, staples, wood, metal pieces. Other related hazards are short weight,
incorrect labels, species substitution, short counts and decomposition.

Chemical hazards: Heavy metals, pesticides, detergents, disinfectants, diesel oil, machine oils,
PSP and DSP .

Biological Hazards:Various bacteria of public health significance, virus and metabolites formed
by bacterial action.

8.2.2 Hazard Analysis

Physical hazards-Analysis

a. stone, sand and mud

● The critical control point is receiving

● This can be controlled by proper verification of the raw material at time of receipt
● Also by washing the raw material thoroughly with water(GMP).

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 b. Glass pieces:

● The CCP is raw material receiving

● This can be controlled by avoidance of glass from premises of seafood processing plants.
● By providing cover to bulbs, tubes etc.

c. Bones, shell pieces, legs, veins, antennae etc

● This is due to improper dressing of the material Rectification – proper dressing of the
material and improving the quality of supervision(GMP)..

8.2.2.1 Health Hazards from Pests

Health hazards from pests:

● Effective pest control programme play a key role in overall sanitation of seafood
processing factories.
● Insects, rodents and birds can easily enter the processing premises and contaminate the
food
● Pest may be divided into 8 groups. Insects and other invertebrates, mammals and birds.
● Relatively few species are encountered in food plant.

Cockroaches Flying insects Rodents Birds

They transfer Flies – good carriers of Health risks from rodents – They transmit
microorganisms to food. bacteria like Salmonella, plague, salmonellosis, microorganisms to foods
Vibrio cholera etc. Jaundice etc. through their droppings.
Objectionable also due to
their offensive smell.

Identification and control measures of pests can be effected at following levels

● Proper plant design and operation

● Design and operation
● Destruction within the plant
● Proximity to a refuse dump increases chances of pests
● By proper design pests can be controlled
● Final line of defense is destruction

Best method of control

Cockroaches Flying insects Rodents Birds

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 Avoid garbage, Avoid Repellents
To eliminate habitat, Removal of breeding chances for entry, Use of and physical barriers
Remove food sources, habitat, Removal of traps and rodenticides
Keeping premises dry, garbage, Fly screens, Air
Sealing of cracks and curtains at entry,
crevices and Application Electrocutors and
of insecticides insecticides

e. Metal f. Filth g.
pieces Decomposition
of fishery
products

From Those engaged Enzymatic

machines, in action,
harvest,transport
From outside, and processing Oxidative
to be are careful. changes,
Metal
detectors – Due to bacteria
final solution

CCP Packaging Raw material,

Receiving-
monitoring
needed

8.2.2.2 Prevention

Prevention

● Checking at the raw material receiving stage

Short weight

● Due to drip loss

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CCP: Packaging

Preventive measures

● Use calibrated balance/weight

● Check each days production for weight

Grade size misrepresentations

● Product size has significant impact on its value

Prevention

● Hourly grade checking

● Calibration of weighing scales

Species substitution

● Affect maintenance of product integrity

Control measure

● Checking at the time of receipt and at the time of packing and labelling

8.2.3 Chemical hazards-analysis

Presence of a chemical may not always be a hazard. The amount of chemical present is
important, Regulatory limits are set for some of these chemicals.
Chemical hazards belong to 3 groups
i. Naturally occurring chemical hazards

● Scombroid (histamine) poisoning

● Ciguatoxin
● PSP / DSP

ii. Unintentionally added chemical

● Pesticides
● Fungicides
● Toxic elements

iii. Intentionally added chemical

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 ● Antibiotics -Aquaculture
● Sulphites -Melanosis control
● Nitrites -Preservatives
● Detergents -Cleaning
● Colouring agents -For bright colour
● Antioxidants -prevention of rancidity

Among the Naturally occurring chemical hazards Histamine production is significant in

scombroid fishes.

8.2.4 Biological hazards-analysis

Biological Hazards-Analysis

It includes:

● Pathogenic bacteria
● Viruses
● Parasites

These hazards come from Raw materials and food processing steps

● Pathogenic bacteria cause illness in human being either by Infection (by swallowing
bacteria) or by Intoxication ( by swallowing free formed toxins). Pathogenic bacteria –
mostly absent in fish caught from off shore waters. Contamination occurs during
handling / processing. Time – temperature conditions, if favorable multiplication is at
faster rate.

Bacteria of public health significance

● E.coil
● Faecal streptococcus
● Staphylococcus aureus
● Salmonella
● Shigella
● V. cholerae
● Parahaemolyticus
● V. vulnificus
● L. monocytogenes
● Camphylobacter
● B. cereus
● Cl. botulinum
● Cl. perfringens

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Chapter 3: Important biotoxin in seafood

8.3.1 Tetradotoxin

This toxin occurs in different animal species such as pufferfish (Tetraodontidae),a goby, Atelopid
frogs Taricha salamanders, octopus and two Japanese shellfishes. This toxin is mainly found in
the liver, ovaries and intestines in various species of pufferfish. The muscle tissue of the fish is
normally free of toxin but there are exceptions. The occurrence is mainly from ovaries and
liver. In octopus, the toxin is present in the posterior salivary venom glands and will be used for
paralyzing the prey. The toxin was isolated as maculotoxin, chemically similar to tetradotoxin
but distinct.
The poisoning leads to neurological symptoms 10-45 minutes after ingestion. It acts by
preventing nerve conduction by an internally specific and reversible blockage of the inward
movement of sodium ions through the cell membrane of an activated neuron. Symptoms are
tingling sensation in face and extremities, paralysis of respiratory system and cardiovascular
collapse. In fatal cases, death takes place within 6 hours.

Tetrodatoxin(TTX) distribution
1. Fish: pufferfish, Fugu(Tetraodontidae) , Goby (Gobius criniger)

2.Amphibians: Newts, Taricha (Salamandridae) frogs, Atelopus varius,

Atelopus chiriquiensis (Atelopidae)

3. Octopus, Haplaochlae maculosa (Octopoda)

4.Shellfish(Gastropoda) Ivory shell, Babylonia japonica; trumpet shell,

Charonia sauliae

5.Star fish , Astropeeton polyacanthus (Echinodermata)

8.3.2 Ciguatera toxin

This poisoning result from the ingestion of fish that have become toxic by feeding on
toxic dinoflagellate. Gambierdiscus toxicus, which is living around coral reef closely attached to
macroalgae. Increased production of toxic dinoflagellates are seen when reefs are disturbed (
hurricanes, blasting of reefs etc.). The toxin ciguatera accumulates in fish that feed on the toxic
algae. Toxin can be detected in gut, liver and muscle tissue by means of mouse assay and
chromatography. The pharmacological action of ciguatoxin is due to cholinesterase inhibition.

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According to Taylor (1988), some fish may be able to clear the toxin from their systems. The
clinical picture varies but onset time is a few hours after ingestion of toxin.

Gastrointestinal and neurological systems are affected due to the toxin. Duration of illness
may be 2-3 days but some may also persist for weeks on even years in severe cases. Death
results from circulatory collapse. The most consistently reported symptoms are

1. Moderate to severe neurological disorders that may persist for days, weeks,or months
and/or
2. Moderate to severe gastrointestinal disorders of relatively short duration
3. In some cases, death due to respiratory failure.

Cigutera may be caused by over 400 species of marine fishes, including many that are highly
priced for food. Five species of organisms namely Gambierdiscus toxicus, Prorocentrum
concavum, P. mexicanium, Gymnodinum sanguineum and Gonyaulax polyedra, produced one or
more toxic fractions which killed mice within 48 hours.

8.3.3 Paralytic shellfish poinsoning

Intoxication after consumption of shellfish is a syndrome that has been known for
centuries, the most common being paralytic shell fish poisoning, PSP is caused by a group of
toxins [saxitoxins and derivatives] produced by dinoflagellates of the genera Alexandrium.,
Gymnodinium and Pyrodinium.

Historically, PSP has been associated with the blooming of dinoflagellates [106 cells/liter],
which may cause reddish or a yellowish discoloration of the water.

The dinoflagellate blooms are a function of water temperature, light , salinity, presence of
nutrients and other environmental condition. However, the precise nature of factors eliciting a
toxic colour is unknown. Water temperature must be >5-8oC for blooms to occur. When
temperature decrease to below 4oC, the dinoflagellate will survive as cysts buried in the upper
layer of sediments.

Mussels, clams, cockles and scallops that have fed on toxic dinoflagellates retain the toxin
for varying periods of time depending on the shellfish. Some clear the toxin very quickly and are
only toxic during the actual bloom, other retain the toxin for a long time, even years.

PSP is a neurological disorder and the symptoms are tingling, burning and numbness of
lips and fingertips, ataxia, drowsiness and incoherent speech. In severe cases, death occurs due
to respiratory paralysis. Symptoms develop within ½ hour to 2 hours after consumption and
victims who survive more than 12 hours generally recover.

Toxin is found in their siphon. Gonyaulox tamarensis& Alexandrium catanella the toxin
is known as saxitoxin. Toxin is heat stables not destroyed by cooking. One gram toxic meat can

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kill 5 people. Estimated lethal dose for man is 1-4mg, respiratory failure followed by death.
Occurrence of this toxin in tropical water is scarce. US FDA limit is 80mg poison /100g.

8.3.4 Diarrhetic shellfish poinsoning(DSP)

Thousands of cases of gastrointestinal disorders caused by diarrhetic shell fish poisoning

(DSP) have been reported in Europe, Japan and Chile [WHO 1984]. The causative
dinoflagellates which produce the toxins are within the genus Dinophysis and Aurocintrum.
These dinoflagellates are widespread which means that this illness could also occur in other parts
of the world. At least seven toxins have been identified including Okadoic acid. Onset of disease
is within 1/2 hour to a few hours following consumption of shellfish which have been feeding on
toxic algae. Symptoms are gastrointestinal disorder and victims recover within 3-4 days. No
fatalities have ever been observed.

8.3.5 Neurotoxic shellfish poisoning(NSP)

Neurotoxic shellfish poisoning [NSP] has been described in people who consumed
bivalves that have been exposed to red tides of the dinoflagellate [Phycodiscus breve]. The
disease has been limited to the Gulf of Mexico and areas of Florida. Brevetoxins are highly lethal
to fish and red tides of this dinoflagellate is also associated with massive fish kills. The
symptoms of NSP resembles PSP except that paralysis does not occur, NSP is seldom fatal.

8.3.6 Amnesic shellfish poisoning(ASP)

Amnisic shellfishpoisoning [ASP] has only recently been identified [ Todd 1990,
Addison and Slenart 1989]. The intoxication is due to domoic acid, an amino acid produced by
the diatom Nitzschia pungens. The first reported incidence of ASP occurred in the winter of
87/88 in eastern Canada, where, over 150 people were affected and 4 deaths occurred after
consumption of cultured blue mussels.
The symptoms of ASP vary greatly from slight nausea and vomiting to loss of equilibrium
and central neural deficit including confusion and memory loss. The short-term memory loss
seems to be permanent in surviving victims, thus the term amnesic shellfish poisoning.

8.3.7 Control of disease caused by biotoxin

The control of marine biotoxins is difficult and disease cannot be entirely prevented. The toxins
are all of non-protein nature and extremely stable (Gill et al.1985). Thus, cooking, smoking,

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drying, salting does not destroy them, and one cannot tell from the appearance of fish or shellfish
flesh whether it is toxic or not.

The major preventive measure is inspection and sampling from fishing areas and shellfish
beds and analysis for toxins. The mouse bioassay is often used for this purpose and confirmatory
HPLC is done if death occurs after 15 min.
To be effective, the monitoring required reliable sampling plans and efficient means of
detection of the toxins. Reliable chemical methods for detection of all toxins are at present
available. Also the toxicity can vary within a growing location of shellfish according to
geography of water currents and tidal activity.

Toxin Tolerance Method of analysis

Ciguatera Control not possible No reliable method
PSP 8.0 ug/100g Mouse bioassay, HPLC
DSP 0.60 ug/100g Mouse bioassay HPLC
NSP Any detectable level/100 g is unsafe Mouse bioassay no chemical method
ASP 20 ug/g demoic acid HPLC

8.3.8 Toxic metals

Toxic metals like Cd, Hg, Se, Pb etc. above normal level affect the safety and marketability of
foods. Many countries have standards. Older ones have higher levels than younger ones of the
same species. Presence of cadmium in cephalopods exported from India is reported. Control by
minimizing pollution & environmental monitoring
Organochlorine pesticides & polychlorinated biphenyls

● OCP & PCBs now common is all environments

Important compounds of concern are

● DDT and its derivatives

● Aldrin and Dieldrin
● Benzene hexachloride
● Polychlorinated biphenyl

It pose special problems in the environment because they are

● Extremely persistent
● They are lipophilic is nature

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 ● Filter feeders like oysters, mussels & clams accumulate these compounds

Permitted levels in seafood

Compounds Max. residue limit (mg/kg)

DDT and its 5

derivatives

Dieldrin 3

PCB 2

Mercury 0.5

Cadmium 3-Crustacean; 4- mollusc

Lead 1-5-crustacean; 1-7 bivalves

Control

● Prevention of contamination and Environmental monitoring

8.3.9 Antibiotic Residues

Widely used in shrimp culture to prevent disease. Usual practice to withdraw the drug in
advance before harvest to enable the animal to clear off the residue. Sometime cultural shrimp
contains high quantities. In-1992 – 1994 Japan rejected many shrimp consignments from
Thailand. Japanese food sanitation act prohibits antibiotic residues in food. From 1994
November onwards US FDA insists testing antibiotic residues in fish

Control:

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 ● Avoid misuse of antibiotics
● Give sufficient time to clear – off the residue

Food additives:

● Food additives including colours are used in various fish & fishery products ( Council
directive 95/2 EC on Food Additives )

Chapter 4: Histamine

8.4.1 Biogenic Amines

Histamine poisoning is a chemical intoxication following the ingestion of foods that

contain high levels of histamine. Historically, this poisoning was called scombroid fish poisoning
because of the frequent association with scombroid fishes including tuna and mackerel. The most
common symptoms are cutaneous such as facial flushing, utricaria, edema, but also the
gastrointestinal tract may be affected ( nausea, vomiting, diarrhea) as well as neurological
involvement ( headache, tingling, burning sensation in the mouth).
Histamine is formed in the fish post mortem by biological decarboxylation of the amino
acid histidine.

The histamine-producing bacteria are certain enterobacteriaceae, some Vibrio sp. A few
Clostridium and lactobacillus sp. The most potent histamine producers are Morganella morgani,
Klebsiella pneumoniaeand Hafinia alviei ( Etratten and Taylor, 1991). These bacteria can be
found on most fish, probably as a result of post-harvest contamination they grow well at 10o C
but at 5o C growth is greatly retarded.

The principal histamine producing bacteria M.morganii grow best at neutral pH, but they
can grow in the pH range 4.7-8.1.The organism is not very resistant to NaCl, but at otherwise
optimal conditions growth can take place in upto 5% NaCl. It should be emphasized that at once
the histamine has been produced in the fish, the risk of provoking disease is very high.
Histamine is very resistant to heat, so even if the fish are cooked, canned or otherwise
heat-treated before consumption, the histamine is not destroyed.

The human body will tolerate a certain amount of histamine without any reaction ingested
histamine will be detoxified in the intestinal tract by at least 2 enzymes the diamine oxidase
(DAO) and histamine N-methyl transferase (HMT) (Taylor, 1986).

Their protective mechanism can be eliminated if intake of histamine and / or other

biogenic amines is very higher if the enzymes are chocked by other compounds as shown in
figure. Other biogenic amines such as cadaverine and putriscine which are known to occur in
spoiled fish may therefore act as potentiators of histamine toxicity. Presumably inhibition of
intestinal histamine catabolism will result in greater transport of histamine across cellular
membrane and into the blood circulation.

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 8.4.2 Control of Disease caused by biogenic amines

Low temperature storage and holding of fish at all times is the most effective preventive
measure. All studies seems to agree that storage at Oo C or very near to 0o C limits histamine
formation in fish to negligible levels. Several countries have adopted regulations governing the
maximum allowable levels of histamine in fishes. e.g.

Regarding limit for histamine in fish

Hazard action Defect action Maximum

allowable level allowable level allowable level
mg/100g mg/100g mg/100g

USA (FDA) 50 10-20 --

EEC ----- 10 20

Toxin Where/when Animals/organisms

produced involved
Tetradotoxin Fish Puffer fish, mostly
ovaries, liver,
intestine
Cignatera Marine algae Tropical/subtropical
fish
PSP Marine algae Filter feeding
shellfish
DSP Marine algae Filter feeding
shellfish

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