Chemistry of meat

Chemical composition of meat

meat can be defined as ―the muscle tissue of slaughter animals‖.

In general, meat is composed of water, fat, protein, minerals and a

small proportion of carbohydrate. The most valuable component from
the nutritional and processing point of view is protein.

 Nitrogenous compounds
Protein 18%
Non Protein 1.5%

 Non nitrogenous compounds

Water 75%
Fat 3%
Carbohydrates 1%
Ash 1.5%
 Water
70-78%
water is a variable of these components, and is closely and inversely related to the fat
content. Adipose tissue contains little moisture; therefore, the fatter the animal, the
lower the total water content of its carcass or cuts. The fat content is higher in entire
carcasses than in lean carcass cuts. The fat content is also high in processed meat
products where high amounts of fatty tissue are used.

They are in following form

―bound water―
―Immobilized water‖
―free water’

The water- holding capacity of the muscle can be decreased by disruptions of muscle
structure. Grinding, chopping, freezing, thawing, salting, degradation of connective
tissue by enzymatic or chemical means, application of other chemicals or organic
additives that change acidity (pH), and heating are treatments that can affect the final
water content of meat products.
 Chemistry of meat

Proteins:
Protein contents and values define the quality of the raw meat
material
and its suitability for further processing. Protein content is also
the
criterion for the quality and value of the finished processed meat
products.
As a source of protein, meat protein is the second largest
component after the water one.

Muscle protein is divided into 3 groups based on their solubility

properties;
 sarcoplasmic protein (30%),
 myofibril protein (55%)
 stromal protein or muscle tissue (15%).
 Sarcoplasmic proteins
are soluble in water and dilute salt solution contained in the
sarcoplasmic.Sarcoplasmic proteins include glyceraldehyde,
aldose, enolase, creatine kinase, lactate dehydrogenase,
pyruvate kinase, phosphorilase, myoglobin, calpain, chatepsin,
extracellular proteins and membrane protein

The sarcoplasma is a soft protein structure and contains

amongst others the red muscle pigment myoglobin. Myoglobin
absorbs oxygen carried by the small blood vessels and serves as
an oxygen reserve for contraction of the living muscle. In meat
the myoglobin provides the red meat colour and plays a
decisive role in the curing reaction. The sarcoplasma constitutes
about 30 percent of the muscle cell.
 Myofibril protein
Myofibrils are solid protein chains and have a diameter of 0.001 –
0.002 mm.

These proteins, which account for the major and nutritionally most
valuable part of the muscle cell proteins, are soluble in saline solution
with a concentration of 1.5% or more.

Proteins are composed of myosin, actin, troponin, tropomyosin, M-

protein, C-protein, Titin, Nebulin and Desmin.

Myosin is the largest component in myofibril and dissolves into the

solution with high ionic strength (greater than 0.3 M).

Myosin has an important functional properties in meat, namely (a) is an

enzyme with ATP-ase activity, (b) myosin form a complex with actin
and (c) myosin form aggregate with each other to form filaments.
 Stromal proteins
Are main constituent of connective tissue that is not soluble in salt solutions but
soluble in alkali or acid treatment. These proteins contain the greatest quantities of
amino avid hydroxyproline.
Connective tissue proteins are found in the following tissues:
1-Sarcolemma elastic sheath that surround the muscle fiber
2- Reticulin fibers: fibrous network around cells that hold tissue together
3- Mitochondria: filamentous or granular organelles of the cytoplasm
Stromal proteins include
1-Collagen: white connective tissue that can be converted into gelatin upon heating
2- Elastin: yellow connective tissue that is almost no destructible
3- Reticulin: connective tissue that resist conversion to gelatin upon moist heating

This protein directly affects meat quality by:

 Lowering the tenderness of meat and it depends on the number of stromal protein
and the degree of cross-linking other stromal proteins.
 Influencing meat emulsion capacity due to this protein is not soluble in water.
 Lowering Water Binding Capacity of meat because of the low content of
hydrophilic and charged amino acids.
 Lowering meat nutritional value.
 Meat colouring
The red pigment that provides the characteristic colour of meat is called myoglobin.
Similar to the blood pigment haemoglobin it transports oxygen in the tissues of the live
animal. Specifically, the myoglobin is the oxygen reserve for the muscle cells or muscle
fibres. Oxygen is needed for the biochemical process that causes muscle contraction in
the live animal.
Factors affecting meat colour:
1. Amount of myoglobin: The greater the myoglobin concentration, the more intense
the colour of the muscle Myoglobin concentration in muscles also differs
a. among animal species. Beef has considerably more myoglobin than pork, veal or
lamb, thus giving beef a more intense colour.
b. The maturity of the animal also influences pigment intensity, with older animals
having darker pigmentation.
c. Sex: bull has more myoglobine than cow
d. Types of muscle:operating muscles as diaphragm have larger percentage of
myoglobin than other muscles
e. Muscle activity:free ranged animals have a higher myoglobin content
f. Dietary iron: low level of iron in the diet lowers the myoglobin level
g. Blood supply; the more sufficient the blood supply the less myoglobin required in
the muscle tissue
h. Oxygen supply: the poor the oxygen supply, the more the myoglobin required
2. Valence of the iron atom 3. Atoms attached to the iron atom
 Fats
Although relatively few in amount, it is the 3rd largest chemical component in the flesh.
One of the disadvantages is the oxidation.
It affects meat processing.
Fat is an important energy source
Fat in meat is generally in triglycerides form.
The composition of triglycerides significantly determines meat tenderness and
roughness.
Fatty acid composition in each species is different and it is also has different effect on
fat properties in each species. Total saturated fatty acids in sheep, cattle and pigs
respectively 53%, 45% and 40%, while the unsaturated fatty acids respectively were
47%, 55% and 60%.
Main types of fatty acids:
1- Saturated (palmitic, stearic)
2- Un saturated (oleic, linoleic, linolenic, arachidonic)
 Carbohydrates
 1%
 Carbohydrates are stored as glycogen

 Stored in the liver and in muscles.

 Affect meat quality

Changes of pH
Immediately post-mortem the muscle contains a small amount of muscle
specific carbohydrate, called glycogen (about 1%), most of which is broken
down to lactic acid in the muscle meat in the first hours (up to 12 hours) after
slaughtering. This biochemical process serves an important function in
establishing acidity (low pH) in the meat.
The so-called glycolytic cycle starts immediately after slaughter in the muscle
tissue, in which glycogen, the main energy supplier to the muscle, is broken
down to lactic acid. The build up of lactic acid in the muscle produces an
increase in its acidity, as measured by the pH. The pH of normal muscle at
slaughter is about 7.0 but this will decrease in meat. In a normal animal, the
ultimate pH (expressed as pH24 = 24 hours after slaughter) falls to around pH
5.8-5.4. The degree of reduction of muscle pH after slaughter has a significant
effect on the quality of the resulting meat
The typical taste and flavour of meat is only achieved after sufficient drop
in pH down to 5.8 to 5.4
The pH is also important for the storage life of meat. The lower the pH, the
less favourable conditions for the growth of harmful bacteria. Meat of
animals, which had depleted their glycogen reserves before slaughtering
(after stressful transport/handling in holding pens) will not have a sufficient
fall in pH and will be highly prone to bacterial deterioration
PSE and DFD
In stress susceptible animals pH may fall very quickly to pH 5.8 –
5.6 while the carcass is still warm. This condition is found most
often in pork. It can be recognized in the meat as a pale colour, a
soft, almost mushy texture and a very wet surface (pale, soft,
exudative = PSE meat). PSE meat has lower binding properties
and loses weight (water) rapidly during cooking resulting in a
decrease in processing yields.
A reverse phenomenon may arise in animals which have not been
fed for a period before slaughter, or which have been excessively
fatigued during transportation and lairage. In these cases, most of
the muscle glycogen has been used up at point of slaughter and
pronounced acidity in the meat cannot occur. The muscle pH24
does not fall below pH 6.0. This produces dark, firm, dry (DFD)
meat. The high pH cause the muscle proteins to retain most of their
bound water, the muscle remain swollen and they absorb most of
the light striking the meat surface, giving a dark appearance.
 Minerals and Vitamins

In addition to protein and fat, meat (beef, veal, pork

and lamb) is a significant source of several other
nutrients.

These include the minerals iron and zinc, Calcium,

magnesium, phosphate, sodium, potassium, sulphar,
chlorine

and most of the B—vitamin complex (B1, B2,

niacin, B6 and B12).
A,C in organ tissue
 Vitamins
 Meat and meat products are excellent sources of the B-complex vitamins
 Lean pork is the best food source of Thiamine (vitamin B1) with more than 1 mg /
100 g as compared to lean beef, which contains only about 1/10 of this amount.
 Meat is a good source of vitamin B12
 On the other hand, meat is poor in the fat soluble vitamins A, D, E, K and vitamin
C. However, internal organs, especially liver and kidney generally contain an
appreciable percentage of vitamin A, C, D, E and K.
 Most of the vitamins in meat are relatively stable during cooking or processing,
although substantial amounts may be leached out in the drippings or broth.
 The drip exuding from the cut surface of frozen meat upon thawing also contains
an appreciable portion of B-vitamins. This indicates the importance of conserving
these fractions by making use of them in some way, for example through direct
processing of the frozen meat without previous thawing (which is possible in
modern meat processing equipment). Thiamine (vitamin B1) and to a lesser extent
vitamin B6 are heat-labile. These vitamins are partially destroyed during cooking
and canning.
 Minerals
The mineral contents of meat include calcium, phosphorus,
sodium, potassium, chlorine, magnesium with the level of
each of these minerals above 0.1%, and trace elements such
as iron, copper, zinc and many others.

Blood, liver, kidney, other red organs and to a lesser extent

lean meat, in particular beef are good sources of iron. Iron
intake is important to combat anaemia, which particularly
in developing countries is still widespread amongst children
and pregnant women.

Iron in meat has a higher bio-availability, better resorption

and metabolism than iron in plant products.
 Enzymes

They play an important role in rigor mortis and

ripening of meat

Enzymes as lipase, amylase, maltose,

phosphates, carboxylase, katepsine and
peroxidase which are important for meat
freshness
 Non protein nitrogenous
Non protein nitrogenous (1.5 % of muscle) are salt water soluble and not
precipitated by trichloroacetic acid
It is composed of nitrogen containing compounds other than protein as follows:
1. creatine: the phosphorylated form stores high energy phosphate
2. Free amino acids: unconnected subunits of protein
3. Nucleotides: basic unit of nucleic acid, ribonucleic acid, deoxyribonucleic
acid, consisting of a base, sugar and phosphate unit
Inosine monophosphate (IMP)
Adenosine triphosphate (ATP)
Adenosine diphosphate (ADP)
Adenosine monophosphate (AMP)
4. carnosine: a dipeptide
5. Anserine; a basic substance found in goose muscle
Extractives are responsible for:
1. Secretion of gastric juice
2. Stimulates digestion and assimilation of food
3. Activate nervous system
4. Responsible for meat aroma
 NON-MEAT INGREDIENTS

chemical substances

 of plant origin

 of animal origin

Other classification criteria for non-meat ingredients are,

whether they are additives or full foods (―food by itself‖) or
whether they have functional properties or not.
Most ingredients are functional, which describes
their ability to introduce or improve certain quality
characteristics. The functional properties of
ingredients include their impact on:
 taste
 flavour
appearance
colour
texture
water binding
 counteracting fat separation
 preservation
Ingredients which are solely functional without any other effect
such as filling or extending the volume of the product, are
normally used in small amounts (e.g. common salt 1,5-3%, nitrite
0.01-0.02%, phosphates 0.05-0.5%, ascorbic acid 0.03%, isolated
soy protein or non-fat dried milk proteins 2%) The criteria for the
utilization of functional non-meat ingredients are:
• safe for consumers, and • improve of processing technology
and/or sensory quality of the products.

In contrast to the exclusively functional substances, there is

another group of ingredients that are not primarily intended for
change of appearance or quality improvements but serve to add
volume to the meat products. They are called meat extenders and
fillers. Their main purpose is to make meat products lower-cost.
Meat extenders and fillers include cereals, legumes, vegetable,
roots and tubers and are used in larger quantities, on average
between 2 and 15%
A-Chemical substances used as ingredients

 Salt (for taste, impact on meat proteins, shelf-life)

 Nitrite (for curing colour, flavour, shelf-life)
 Ascorbic acid (to accelerate curing reaction)
 Phosphates (for protein structuring and water binding)
 Chemical preservatives (for shelf-life)
 Antioxidants (for flavour and shelf-life)
 Monosodium glutamate MSG (for enhancement of flavour)
 Food colouring substances (synthetic and of plant origin)

Chemical additives have exclusively functional properties,

they are used in small amounts usually below 1% (with nitrate
as low as 0.05%). Only salt is in the range of 2% (with up to
4% in some fermented dried products).
 B- Non-meat ingredients of animal origin
Ingredients of animal origin are not commonly applied but may be
useful
for specific meat preparations. They all have functional properties
(except whole milk), in particular improvement of water binding and
prevention of fat separation during heat treatment. Apart from their
functional properties, some of them can also be considered meat
extenders, as mentioned below.
 Milk caseinate (90% protein; used in small quantities (2%);
have functional water and fat binding properties)
 Whole milk or non-fat dried milk (=skim milk) (sometimes
used in indigenous meat preparations as a protein extender)
 Gelatin (binding properties and meat extender)
 Blood plasma (predominantly binding properties)
 Eggs (extender and binding ingredient for meat pieces and fried
sausages)
 Transglutaminase* (exclusively binding properties)
 C. Ingredients of plant origin
 All spices:They are predominantly functional and used in small
quantities to provide or add flavour and taste to meat products.

Binder
functional substances of plant origin with high protein content to
increase water binding and fat retention, in particular in intensively
heat treated products
• isolated soy protein (90% protein)
• wheat gluten (80% protein)
They are not extenders in the first place due to the low quantities added
(approx. 2%), but act through their high quality proteins that are
instrumental in water binding and protein network structuring. On the
other hand, some substances with little or no protein level, like
starches and flours mentioned above under ―fillers‖, can bind water
and fat by means of physical entrapment and could also be considered
―binders‖.
Meat extenders / Plant products with high protein
content are
􀂾 Soy flour (50% protein)
􀂾 Soy concentrate (70% protein)
Other food legumes (beans, peas, lentils), used for
special products only.

These cheaper plant proteins “extend” the more

expensive meat proteins, resulting in acceptable
overall protein contents of lower cost meat products.
Extenders are added in sizeable amounts that increase
the bulk of the meat products, but this may also alter
their quality.
 Fillers

Carbohydrate products with low protein content (usually added

in quantities of 2%-15%). These are the typical fillers. Apart from cost
reduction and adding to volume, some flours and starches belonging to
this group of fillers also act to some extend as binders. This property
serves important functions such as increasing water binding for more
juiciness or fat binding for improved texture.

􀂾 Cereal flours from wheat, rice and corn

􀂾 Starches from wheat, rice, corn, potato and cassava
􀂾 Breadcrumbs
􀂾 Rusk (derived by mixing and baking wheat flour)
􀂾 Cereals to be added without milling, e.g. rice, corn
􀂾 Roots and tubers, e.g. cassava, sweet potato
􀂾 Vegetable and fruits, e.g. onions, bell pepper, carrots, green
vegetables, bananas
􀂾 Polysaccharides (Hydrocolloids):
Chemical Residues in
Meat
Introduction
Veterinary medicines are substances used to treat and prevent disease in
animals and thus have the potential to enter the food chain through their
occurrence as residues in food.

Animals may also be exposed to a range of other chemicals such as

pesticides and environmental contaminants (organohalogens, chemical
elements, mycotoxins, etc.) occurring in air, soil, water or in feed.

A further category of substances of potential concern is growth promoting

agents and non-approved veterinary medicines that are prohibited for use
in food-producing animals.

It is of critical importance that residues of veterinary medicines,

contaminants and prohibited substances are either not present in animal
products destined for the human food chain, or are present at such a level
that adverse effects on the health of consumers cannot occur.

Detection of these unwanted residues presents a new challenge to meat

hygienists.
Chemical residues are the traces of a chemical or its breakdown
products that remain in or on treated product after a particular time.

Maximum residue limits (MRLs)

The maximum residue limit (MRL) is the maximum concentration of a

chemical residue that is legally permitted to be present in a food. This
concentration is expressed in milligrams per kilogram (mg/kg) of the food.

The detection limit, lower limit of detection, or LOD (limit of detection)

is the lowest quantity of a substance that can be distinguished from the absence
of that substance (a blank value) with a stated confidence level (generally
99%).The detection limit is estimated from the mean of the blank, the standard
deviation of the blank and some confidence factor
Residues may include

1-Therapeutic products:

a- Antimicrobials
b- Antiparasitic
c- Hormones

2- Environmental contaminants: including herbicides, heavy

metals and fungicides.

3-Food additives and food processing intoxicant

4- Natural toxins (Mycotoxin)

 Antimicrobials

The most numerous and most frequently used drugs in

this group are the antibiotics and sulphonamides.

An antibiotic is a chemical substance, produced wholly

or partly by a microorganism (usually a fungus or
bacterium) which has the capacity to inhibit the growth
of or to kill bacteria.

These drugs can be used therapeutically in short courses

of treatment to control disease in animals or, at lower
concentration but over a longer time, to promote growth.
The latter use occurs most frequently in young calves
and poultry.
Antibiotic residues are considered undesirable for several reasons

 They produce unsightly lesions when administered by injection

 During meat inspection all carcasses with injection sites should be

retained and judgments made according to case history, the time of
treatment and laboratory results.

 Antibiotics may interfere with further food processing if this

depends on fermentation reaction.

 They may cause allergic reactions in sensitized consumers.

 A small number of antimicrobials are suspected of having

carcinogenic properties.

 Bacterial resistant
Tests for antimicrobial agents

1- Detection of residual antimicrobial activity. The basic microbiological

method is the four plate test (FPT). It is an agar diffusion test. Meat samples
are applied to four plates of agar medium, three of which are inoculated with
Bacillus subtilis spores at pH 6 , 7,2 and 8. Diffusion of the active antibiotic is
detected by the formation of zones of inhibition on one or more plates after
overnight incubation. The reliability and sensitivity of the test is monitored by
applying 6 mm-diameter filter paper discs containing standard quantities of
known antibiotics in each turn.

2- High voltage electrophoresis (HVE) bio autography. Two gels, agar and
agarose are prepared, a piece of meat is placed on each and the antibiotic is
allowed to diffuse into the medium. The high voltage is passed through the
medium for a period of 2.5 hrs. The plates are then over-laid with media
containing sensitive species of bacteria similar to those in the FPT and
incubation is carried out overnight. The antibiotics inhibit the growth of
bacteria over the area in which they are concentrated.

3- Other methods of detection includes LC-mass/mass- HPLC.

 Hormones
Hormones have been used for a variety of therapeutic and growth –modifying
purposes in animals.

The most commonly used hormones are:

 Natural sex steroid hormones - Oestradiol, progesterone, testosterone.

 Synthetic steroid androgens - Nandrolone, norethandrolone,

nortestosterone, phenylpropionate, ethinyloestradiol, laurate.

 Synthetic non-steroidal oestrogens -Stilbene estrogens

(diethylstilbestrol) (DES), hexoestrol), zeranol, trenbolone acetate

 Synthetic steroidal progestens - Melengestrol acetate (MGA).

 Peptide hormones - Growth hormone (GH), growth hormone-releasing

factor thryotrophin-releasing hormone (TRH).

 β-Adrenoceptor agonists (beta-agonists) - clenbuterol, cimateratol.

When the ban was first introduced, producers had access to other
growth promoting hormone implants ( trenbolone, zeranol, natural
hormones) which gave better responses at slightly higher costs.

Tests for growth promoting hormones have been implemented:

 In the live animal which is tested on farm, blood or feces are the
most convenient samples to collect.
 At slaughter, blood, rectal feces, liver, kidney and muscle can be
obtained from all animals.
 Screening tests for residues of hormonal growth promoters are
based on immunoassays. These tests are rapid, sensitive, selective
and cost-effective.
Effect of hormones
May be carcinogens
 Anthelmentic
Used to remove internal parasites such as liver fluke and nematodes
are important in animal production systems.

The salicylanide flukecides, oxyclosanide, closantel and rafoxanide

are active against Fasciola hepatica.

Thiabendazole was the first highly effective broad spectrum

anthelmintic and is being used in the treatment of nematode infections.
In general these therapeutic drugs are used to control infections in
farm animals therefore they are unlikely to be administered close to
slaughter.

A number are administered by injection which can be irritant, and

many animals require to be examined so that the active drug, if any
residue remains, can be identified.
 Pesticides
Pest control chemicals must be toxic to some living organisms to
fulfill their role. Depending on the pest being controlled they may be
termed insecticides, fungicides, etc. The insecticides that are directly
applied to food animals and the anthelmintics are regarded as the
most important subgroups
Types:
Organochlorine
Organophosphate
Pesticides are detected by chemical techniques. In the laboratory,
spectrographic methods of pesticide analysis using color producing
reactions were the first to reach sensitivities at the ppm level but these
methods have been replaced by chromatographic techniques.
Effect:
Carcinogenic, developmental and growth problems, Liver and kidney
failure
 HEAVY METALS:
Excessive intakes of heavy metals in food have caused intoxications in man.

Sources:

 Contaminated cereals or by accidental additions during processing.

 Soils naturally high in the element or through environmental contamination
from local industry and are cumulative in animal tissues.
 Feeding grain treated with the toxic metal
 previous use in paints, etc.

These toxic chemicals are detected by atomic absorption spectrometry.

Lead:
Acute cases are rare and occur most commonly after ingestion of lead-
containing paint.
In these the highest concentrations are found in liver and kidney.
Acutely- affected animals should be detected during ante-mortem inspection.
At post-mortem the muscle of acutely-poisoned bovines is unusually pale.
Arsenic:
Fed, forage or liquids contaminated with arsenical herbicides, rodenticides
or insecticides.
Arsenic-containing compounds have been used for parasite control, and for
the treatment and control of swine dysentery, but these have largely been
removed from the market.
Only the liver approaches the hazard level for man
Shellfish can accumulate particularly high concentrations if taken from
polluted waters. Bottom feeders from these areas also accumulate the metal
but free- swimming fin fish are less affected.

Mercury:
widely in agricultural and in veterinary medicines.
Although mercury is extremely toxic, cases of poisoning are rare.
They have been most frequently associated with feeding to animals of seed
grain treated with mercury-containing dressings to prevent fungal growth.
High concentrations may also occur when industrial pollution contaminates
grazing areas or when sewage sludge is used intensively as a fertilizer.
Cadmium:
cause kidney failure. In farm animals the greatest concentrations occur in kidney and
liver. The origin of these residues may be sewage sludge or organocadmium fungicides.

Copper:
Copper-supplemented feeds
Copper-supplemented feed prepared for pigs has accidentally been fed to sheep and led
to chronic copper poisoning in this species

Selenium:
Selenium is an essential element for animals and man. In some cases acute selenium
poisoning may occur in cattle grazing pasture that contains plants which accumulate
this element

Fluorine:
Cases of fluorosis have been reported in cattle grazing pasture contaminated with
industrial discharges.
This chronic disease is associated with staining of the teeth and excessive wear and
degenerative changes in the skeletal system and internal organs.
It has not been associated with illness in man.
 Dioxins ( sources)
• Dioxins are mainly by products of industrial processes but can also result from natural
processes, such as volcanic eruptions and forest fires
• Dioxins are by products of a wide range of manufacturing processes including
smelting, chlorine bleaching of paper pulp and the manufacturing of some
herbicides and pesticides. uncontrolled waste incinerators (solid waste and hospital
waste) are a major source of environmental realse.
• Although formation of dioxins is local, environmental distribution is global..
The highest levels of these compounds are found in some soils, sediments and
food, especially dairy products, meat, fish and shellfish. Very low levels are found
in plants, water and air.
 Dioxins ( Expousure)
•Short-term exposure of humans to high levels of dioxins may result in skin lesions,
such as chloracne and patchy darkening of the skin, and altered liver function
•Long-term exposure: impairment of the immune system, the developing nervous
system, the endocrine system and reproductive functions
•Chronic exposure of animals to dioxins has resulted in several types of cancer.
However, TCDD does not affect genetic material and there is a level of exposure
below which cancer risk would be negligible.
•The developing fetus is most sensitive to dioxin exposure and also the newborn.
•individuals at risk: 1- high consumers of fishes and 2- occupations (e.g., workers in
the paper industry, in incineration plants)
 Prevention and control of dioxin
exposure
•Proper incineration of contaminated materials ( 850-1000oC)
•Prevention or reduction of human exposure is best done via source-
directed measures, i.e. strict control of industrial processes to reduce
formation of dioxins as much as possible.
•protecting the food supply is critical ( 90% of human exposure is due
food).
•Food contamination monitoring systems.
 NATURAL TOXINS

Mycotoxins:

 Mycotoxins are products of toxigenic moulds (fungi) growing in food

and foodstuffs.
 These agents have caused many problems in livestock
 The potential for residues in meat, poultry or dairy products is a cause
for public concern.
 The risk to human health from direct consumption of contaminated
grain is much higher than that arising from animal products.
Aflatoxins

Produced by Aspergillus flavus and Aspergillus parasiticus.

There are four major types of toxin labeled AFB1, AFB2, AFG1, and
AFG2.

AFG1 is the most commonly produced and the most toxic.

Much of the ingested toxin is excreted within 24 hours and excretion is

almost complete within 96 hours after ingestion ceases.

Liver and kidneys retain detectable quantities for longer periods than
other tissues.

Dairy products are considered to be the most vulnerable to residue

accumulation (Shank, 1981)

Transmission of toxic amounts into human food through meat and meat
products does not appear likely.
Ochratoxins

 Produced by some Penicillium spp. and some Aspergillus strains.

 Ochratoxin A is the most common and the most toxic to mammals,

birds and fish.

 The kidney is the primary target organ, but liver damage has also been
recorded at high concentrations.

 It has been shown to be a teratogen in laboratory animals but has not

been proved to be carcinogenic (Busby and Wogan, 1981).

 The highest risk for consumers is the potential for residues to

accumulate in kidney.

 Lower concentrations occur in liver, fat and muscle.

Detection

 Immunoassay kits

 Positive animals are identified, they should be retained on a toxin-free diet

for 4 weeks prior to slaughter to ensure that the levels in kidney have
decreased.

 In poultry, residues have been detected in liver, kidney and muscle but not in
eggs. A 48-hour withholding period is sufficient to clear muscle.

 In ruminants, ochratoxin A is detoxified in the rumen; hence accumulation in

their tissues is highly unlikely.

 Biotoxins

 Biotoxins are toxic substances of biological origin accumulated in certain marine

species.

 They classified into two groups

1- Toxins which are produced by certain marine algae like paralytic shellfish,
diarrhetic shellfish, neurotoxic shellfish, amnesic shellfish poisoning and ciguatera.
Where the toxins are produced by certain species of naturally occurring algae when
bloom under favourable conditions
• Filter-feeding molluscan shellfish (clams, oysters, mussels and
scallops) accumulate the toxins when utilizing toxic algae as a food
source. As the conditions become less favourable, the bloom
subsides and with time, shellfish rid themselves of toxin and are
once again safe to eat.

2- Toxins which are naturally occurring in certain species of fish that do

not involve marine algae, like Tetrodotoxin, Gempylotoxin and
Tetramine
 Melamine
 Melamine is an organic compound commonly used to
produce various products, including dishes, plastic
resins, in fertilizer

 The average concentration of melamine in food from

approved industrial uses is estimated to be less than
0.015 parts per million (USDA, 2009).

 These levels of melamine in food are extremely

minute and do not pose a public health concern.
 Food processing toxicants
 During the processing of foods, products may be produced that, if present in
large amounts, could potentially adversely affect health.

 For example, cooking certain meats at high temperatures creates chemicals

that are not present in uncooked meats.

 A few of these chemicals may increase cancer risk, such as polycyclic

aromatic hydrocarbons and heterocyclic amines.

 Another example is when nitrates and nitrites react with secondary amines
to form nitrosamine.

 Nitrosamines are mutagens which have been linked to cancers.

 Nitrates and nitrites are used to preserve meats and contribute to prevention
of growth of Clostridium botulinum, the bacterium responsible for
producing the highly potent botulinum toxin.
a. Polycyclic aromatic hydrocarbons

Polycyclic aromatic hydrocarbons (PAH) are produced when any

incomplete combustion occurs.
Thus, they are found in cooking oil fumes, smoked foods and foods
cooked at high temperature.
Most PAHs are not carcinogenic, although a few are, for example benzo (a)
pyrene.
They appear mainly in meats cooked during high temperature grilling.

Microwaving does not produce PAHs and foods other than meats contain
negligible amounts of PAHs.

Foods low in fat, or cooked beneath the source of heat, contain many fewer
PAHs, so the type of food cooked and the method of cooking are important
determinants of PAHs.

Breathing air containing PAHs can occur in the workplace of smokehouses.

Eating grilled or charred meats, meats and processed or pickled foods
increases an individual's exposure to PAHs.
Heterocyclic amines Heterocyclic amines (HCAs)

 Carcinogenic chemicals formed from the cooking of muscle

meats such as beef, pork, fowl, and fish.

 HCAs form when amino acids (the building blocks of

proteins) and creatine (a chemical found in muscles) react
at high cooking temperatures.

 Some 17 different HCAs have been found, resulting from

the cooking of muscle meats that may pose human cancer
risk.
C. N-nitrosamines

Approximately 300 of these compounds have been tested and 90% of

them have been found to be carcinogenic in a wide variety of
experimental animals.

Dimethylnitrosamine (DMNA) (also called N-nitrosodimethylamine or

NDMA) is a member of a group of chemicals known as nitrosamines
which are recognized as cancer-causing substances.

Cured meats can contain nitrosamines because meats contain amines and
sodium nitrite, a source of nitrosating agents added to cured meats as a
preservative.
Of all the cured meats,bacon has received the most attention.

Removal of sodium nitrite would prevent nitrosamine formation, but it

might also increase the risk of botulism poisoning.

Sodium nitrite and sodium chloride together are particularly effective

against Clostridium botulinum.
 Detection, evaluation and determination
techniques and equipment
1. Microbiological Assays:
The microbiological methods used for detecting antimicrobial residues
in foodstuffs and based on inhibiting microbial growth, microbial
receptor activity and enzymatic reactions. Microbial inhibition assays
involve culturing a microorganism from a standard strain, usually
Bacillus stearothermophilus, Bacillus subtilis, Bacillus cereus,
Micrococcus luteus, Escherichia coli.

1. Immunological Techniques
These methods are based on the interaction antigen-antibody which is
very specific for a particular residue. The most common technique is
enzyme-linked immunosorbent assay (ELISA). ELISA kits are available
for a specific residue such as sulpametazine or a group of related
compounds such as sulphonamides. ELISA kits have shown good
performance for the analysis of antibiotic residues in meat
2. Biosensors
Different types of biosensors have been developed in recent years as an
alternative approach to screen residues in animal products. In general The
biosensor is an instrument combining, in close contact, a recognition element
with an antibody/antigen pair, a receptor and its specific ligand, or even living
cells and an analyt that binds specifically to them. The biochemical signal of
biosensors are converted by a transducer into an electronic signal. Then, these
signals Public Health Importance: Worldwide national and are processed by a
microprocessor that gives the final result .
3.High Performance Liquid Chromatography (HPLC) HPLC allows the
qualitative and quantitative detection of multi-residues in meat and fish
products

4. Charm II Technology Charm II technology is a new technique which is

rapid, comprehensive and semi quantitative testing system capable of detecting
residual compounds. This technology can provide the desired selectivity and
sensitivity. The use of Charm II along with HPLC separation provides an
excellent method for the detection and identification of individual chemical
and biological residues in animal tissue.
 Hazard Control and Prevention
Successful chemical control program should include:

• Train employees to follow safe handling and application procedures

for sanitation, maintenance or pesticides chemicals.

• Store chemicals in designated areas away from food, ingredients,

packaging and food contact surfaces.

• Make it standard practice for staff, after maintenance, to properly

clean and remove all chemical residues from food contact surfaces.

• Do not use excessive grease or lubricants on equipment. Regularly

re-evaluate all procedures to ensure they effectively remove chemicals.

• Ensure chemical containers and measuring tools are clearly labeled

or color coded, and that they are used only for chemicals.
• Use designated tools for handling allergens and scheduling
products using allergens last in the production cycle.

• Store allergens to prevent cross-contamination with other

ingredients (ex: tightly close containers, use separate storage room,
or ensure adequate physical separation).

• Receive incoming materials and raw ingredients from reputable

suppliers that effectively control chemical hazards, so you prevent
the introduction of these hazards in your plant.

• Ensure restricted ingredients and additives are correctly measured.

Regularly re-evaluate all recipes to ensure they meet the Food and
Drug Act and its regulations.

• Follow good storage practices (ex: uncontrolled moisture levels

during grain storage can produce mycotoxins).
Identification of animal
species
Identification of animal species include
• Identification of the stamp (shape & color)
• Differentiation of carcasses of slaughtered animals
• Characteristics of muscles
• Characteristics of fat
• Anatomical variations of organs and bones
• Characteristics of hair and wool
• Physical and chemical means for identifications of
meat and fat
• Biological means for identification of meat
 1. Stamping

1- young animal

2- old animals

3- Imported

4- Imported slaughtered
at the country of origin
 2. Differentiation of carcasses
Differentiation between bull and cow
Item Bull Cow
• M. Of neck, shoulder well developed ill developed
& fore quarter
• External genitalia Removing testicles & Removing udder
spermatic cord leave leave triangular space
a ring covered with fat with large amount of fat

• Pelvic cavity Narrow Wide

• Gracilis M. Triangular Bean shape
•Bulbo cavernous M. Present absent
•Root of penis Present absent
•Pubic tubercle Large & rounded Small &flattend
 Differentiation between sheep and goat
Item Sheep Goat
• Back Rounde & well fleshed Sharp & little flesh
• Thorax Barrel- shaped flattened
• Tail fatty Thinner
•External lacrimal notch present absent
•Spinal process of cerv. Vert. Broad & blunt Pointed & Sharp
•Sacral Vertebrae 4 3
•Sacral border Thick Thin
•Pelvic opening wide narrow
•Hair 7 wool some wool coarse hair attached to carcass
 Differentiation between Cat and Rabbit
Item Cat rabbit

• Head Rounded, 4 canines elongated, no canine.

• Radius & ulna Separated except united
or tibia & fibula at extremities
• Liver 7 lobes 4 lobes
• kidney pinkish brown chocolate
•Tail long, fleshy short
•Thorax long short
•Fore arm heavy, muscular light, bony
•Xyphoid cartilage short, fibrous cartilaginous
•Fat white Honey yellow
•Meat Pale pale-red to grey-red
•Transverse process Directed downward & fore with forked end
of lumber vertebrae with pointed end
Camel
Very long extremities and neck
Chest pad and elbow pad
Single or double hump
Short trunk & rounded belly
Soft and greasy fat
• Horse
Long extremities, neck and thorax
Massive development of pectoral M.
18 pairs of ribs
Oily golden yellow fat
3,4. Characteristics of meat and
fat of various animals
 Cattle
Meat Fat
• Red to reddish brown • Colorless or
• Firm in consistency yellow ,
creamy & firm
• Meat intermixes with fat
(Marble appearance) • Become hard
• Young cattle (6-15 M) after chilling
has fine fibers, pale to
brick in color & slightly
intermixed with fat.
 Buffalo
Meat Fat
• Fresh light to dark • Pure white
brown • Dry, not oily, sticky
when rubbed between
• M.F. coarse, fingers
become tough and • Renal fat is poorly
not easy to cut developed, of dull
after cooking appearance & and
• Not intermixed shrinks when cooked
with fat
 Camel
Meat Fat
• Rose- red, coarse • Creamy yellow,
M.F. soft & greasy
• Accumulated in
• Very tough on
hump and
cooking
sternum
• M. Not intermixed
with fat
 Horse
Meat Fat
• Dark red to brown • Soft, oily,
• On exposure to air a quire golden yellow
bluish or even black luster • Remain soft
• M. Not intermixed with fat after chilling
• Rich in glycogen
• C.T is strongly developed
 Veal
Meat
• Pale grey to
Fat
grayish red • Milky and
• Fine fibers, not white
intermixed with
fat
• Bones are soft and
bluish red
 Mutton
Meat Fat
• Light red to dark red • Pure white
• Fine texture & hard
• Not intermixed with fat
• In fatty animals fat is
found in between groups
of M., S/C& kidney
capsule
• Slight ammonical odor
 Goat
Muscle Fat
• Light red to dark red •S/C fat is little
• Buck urine odor and sticky
• Sticky •Kidney has a
thick fat capsule
• During skinning hair
attached to meat
 Pork
Meat Fat
• White grey to pale •White, soft, greasy
grey & granular
• Fine fibers, •S/C fat is very
intermixed with fat much
•More preferable in
• Become white on industry ( easy in
cooking emulsification)
• high cholestrol
 Fetal Flesh
Meat Fat
• Very soft & flabby •Undeveloped,
• Watery & gelatinous jelly like or
• High glycogen content gelatinous
 Dog
Meat Fat
• Dark red • White to whitish
• firm grey, oily &
greasy
• M.F. slightly
intermixed with fat
 Rabbit
Meat Fat
• Pale red to grey red to • White yellow
grey concentrated
• Fine fibers, not in body cavity
intermixed with fat
 Poultry
Meat
• Fine& firm fibers, not intermixed with fat
• White Chicken & turkey
• Dark red duck, geese & pigeons
5. Anatomical variations in
organs and bones
Cattle
Camel
Sheep
Pig
6. Characteristics of hair
and wool
Physical and chemical means for
identification of meat
a. Fat crystal test:
2.5 melted fat + 7.5 ml (mix. Of alcohol & ether
1:1) leave in refrigerator 12 hr.
Examine cystals under microscope

Beef fat Lard

Sun shape with pointed end Brush shape with blunt edge
b. Physical test for lard:
Melt fat in container or spoon, then leave
to solidify
Beef fat solidify in smooth surface
Lard solidify in the form of depression
surrounded with concentric rings.
c. Sulphoric acid test:
Addition of H2SO4 during cooking of horse meat
repulsive odor of horse stable due to volatile
F.A with formation of yellow oil globules on
the soap surface.
d. Ehirilich test
soaking horse flesh in formaline for 48 h.
Result in intensive characteristic odor resemble that
of roasted geese.
e. Glycogen test
5 g minced meat + 5 times its volume D.W
Boiling
15 min (pure meat
30 min. (meat products)

Cooling then filtration

Small part of filtrate + drops of

lugols iodine

Dark brown Bluish Starch

PPT of starch by conc. Glacial acetic
acid, filter and apply the test (filtrate
+ lugols)
glycogen
Dark brown Dark yellow
glycogen starch
 +ve glycogen test

Liver Fetal flesh

Horse Can not be det. Due to
protein is not well
develop. So no Ag-Ab
Histological sec. reaction.
PPT. test
 8. Biological means for identification of
meat
1. Immuno-chemical method
a. Precipitation test Tube ring test
Agar gel diffusion test

b. Hemagglutination test
c. ELISA
d. Complement fixation test
2. Electrophoresis fractionation of proteins
3. Polymerase chain reaction (PCR) identification
of DNA recovered from meat samples
4 gas liquid chromatography
5. Histo-chemical method
 Precipitation test
• It is a biological test for differentiation of
meats of various animal species.
• The inspector may identify certain cut of
meat but can not identify the meat if it is
present in the form of minced meat or
sausage.
Significance
1. It is valuable in identification of fresh, frozen,
pickled, smoked and cooked meat as well as bone.
2. It is accurate (decisive test).
Disadvantage
1.The result is doubtful in case of smoked or cooked
meat, if the temp. in its center exceeds 70ْ C.
2. It is of no value in differentiation of meats of closely
related animals as horse and donkey.
• Idea of the test
It depends on Ag (unknown meat)-Ab
reaction
• Procedure
1. Preparation of unknown meat extract
2. Preparation of known antiserum
3. Reaction
Preparation of unknown meat
extract
M.E. must be free fat and salt
• Fat extraction
50 gm minced meat + 100 ml ether &
chloroform 1:1 shake every 0.5 hr.
for 24 hr. to extract fat discard the
mixture wash the meat by addition
of normal saline
• Desalting (salt dissolving)
50 gm mined meat + 50 ml dist. H2O shake
for 5 min., pour the supernatant fluid and
repeat washing for several times
• Meat Extract preparation
50 gm minced meat free from fat & salt mixed
with 100 ml sterile saline keep in
refrigerator for 12 hr. at 2-3ْ C filter by
ordinary & bacterial filter.
 Protein content investigation
a. Nitric acid heating test:
2 ml M.E + 1 drop 25 % nitric acid (sp. G.
1.153) heating
If marked cloudiness and precipitate indicate
high protein The meat extract must be
diluted by sterile saline, till the M.E. give
only slight turbidity. Slight turbidity after 5
min. means that the ratio between protein &
normal saline is 1: 350
b. Shaking test:
• 2 ml M.E. Shake well till formation
of foam, the longer the time of
formation of foam, the suitable
concentration of protein for biological
examination (1: 350).
• The final diluted M.E. examined for pH.
Which must be 7 (neutral).
• The M.E. is now ready for precipitation
test
Preparation of known antiserum by
rabbit injection with known M.E.
The M.E. of known animal is prepared & injected I/V into a
rabbit in the following ways
0.2 ml 3 times with an interval of 2 days
0.4 ml 2 times with an interval of 2 days
0.6 ml 2 times with an interval of 2 days
0.8 ml once after elapse of 24 hr.
1 ml once after elapse of 24 hr.
Rest the animals for 14 days
The rabbit is inoculated with 0.2 ml of serum from
animal in question for 2 times with 24 hr. interval.
Bleeding of the rabbit (is better after a
period of fastening to obtain clear
antiserum of known animal)
Rabbit antiserum should be tested toward
different kinds of M.E. to ensure specificity.
 Technique of the test
• Tube ring test
Using aggl. Tube, add equal amounts of
suspected M.E. and antiserum
Suppose the meat in question is horse meat
H.A.S B.A.S H.A.S B.A.S N.R.S H.A.S

S.M.E S.M.E H.M.E B.M.E S.M.E Saline

Positive control Negative control

Positive result is indicated by distinct precipitation ring within

5 min.
• Agar gel diffusion test
M.E M.E
Nobel agar

Pure Horse B.A.S H.A.S

B.M.E H.M.E B.M.E H.M.E

M.E M.E

H.A.S
B.A.S
Pure Beef

B.M.E H.M.E B.M.E H.M.E

M.E M.E

Mix B.A.S H.A.S

B.M.E H.M.E B.M.E H.M.E

• Histo-chemical method
For detection of plant protein using calleja stain
Muscle green with striation & dark brown
nuclei on the periphery
Soya protein Tuft or network of un special
structure without striations or nuclei & dark
green color
Spices Brownish
C.T. Blue
Fat Empty vaccules