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SOLID-STATE FERMENTATION IN ANIMAL NUTRITION

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 Volume 3 – Issue 12
Online ISSN: 2582-368X

SOLID-STATE FERMENTATION IN ANIMAL NUTRITION

Article Id: AL202202
1
Rashika Srivastava*, 1Parul Rana and Shubham Singha2
1
Animal Nutrition Division, ICAR-National Dairy Research Institute, Karnal-132001, India
2
Veterinary Gynecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal-
132001, India
Email: [email protected]

S
olid-state fermentation (SSF) is a fermentation process occurring in a solid matrix in
the presence of a little or negligible amount of free water, provided that the substrate
must have sufficient moisture to support the growth and metabolism of microbes. It
can be delineated as a process of the growth of micro-organisms on a solid substrate without
a free-flowing aqueous phase.The solid matrix under consideration might be a reservoir of
nutrients or merely a support infused with nutrients allowing microorganisms to grow.It is a
fermentation method used by industries like the pharmaceuticals, food, textile etc., to produce
biologically active secondary metabolites from microbes; and is emerging as a potential
alternative to submerged/liquid fermentation(Nigam and Pandey, 2009).The advantage of
SSF is accountable to the fact that it brings the microorganisms under cultivation in close
proximity of the substrate and hence maximal substrate concentration is achieved. It mimics
natural microbiological processes of fermentation like composting and ensiling. SSF provides
a favourable habitat to microbes, replicating their natural environment they grow in, hence is
highly preferred to grow and produce useful value added products in biotechnological
industries. Processes where SSF finds application are:

Research in the field of SSF

dates back to 1960–1970,
when production of protein
enriched cattle feed by SSF
was done utilizing agro-
industrial residues, thus
offering a perfect solution for
the development of nutritionally enriched roughages for livestock, and simultaneously
reduction of air pollution. Recently, it is gaining more popularity in the solid waste
management, biomass energy conservation and biotechnology industries (Pandey, 2007).

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Rice straw, sugarcane bagasse, wheat straw, rice hulls, and corn cobs are among the
agricultural wastes utilised as SSF substrate (Gonzalez et al., 1993).

Steps in Solid State Fermentation

SSF is normally multistep processes involving the following steps:

i. The process is initiated with the pre-treatment of substrate raw material by

mechanical, chemical or biochemical processing to increase the surface area and
increase nutrient availability.
ii. Deposition of a solid culture substrate, such as rice or wheat bran, on flatbeds after
seeding it with microbes for fermentation
iii. The substrate is then left in a room with temperature-control system for some
days.
iv. Hydrolysis of primarily polymeric substrates, e.g., polysaccharides and proteins.
v. Utilization (fermentation) of hydrolysis products.
vi. Separation and purification of end products.

Substrate (Bran, expeller cake)

Energy Mechanical and heat pre-treatment

Pre-treated substrate

Energy, water
inoculum Solid fermentation

Fermented product
(biocatalyst)

Nutrition and Animal Bio-protection for

Biofuel
human health feed grain crops

Fig.1: Solid-state fermentation process for production biomolecules and value-added

products (Srivastava et al., 2019)

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Applications of SSF

Solid-state fermentation has emerged as a potential methodology for the production of

microbial products such as feed, fuel, industrial chemicals, and pharmaceutical products.

It is widely employed to manufacture several enzymes, organic acids, which are

subjected to extraction and purification, employed to produce different products.

It can also be used in bioremediation and bio-leaching.

Application of SSF in production of certain active secondary metabolites are listed below.

Product Use Source Substrate

Pharmaceuticals
Zearalenone Growth promoter Fusariummoniliforme Corn
Bacterial
Insecticide Bacillus thuringensis Coconut waste
endotoxin
Sugarcane
Penicillin Antibiotic Penicilliumchrysogenum
bagasse
Cephalosporin Antibiotic Cephalosporiumarmonium Barley
Oxytetracycline Antibiotic S. rimosus Corn cob
Immuno suppressive
Cyclosporin A Tolypocladiuminflautum Wheat bran
drug
Enzymes
Aspergillus niger,
Gingelly oil cake, coconut cake, babassu oil
Lipase Candida rugosa,
cake,
Penicilliumrestrictum
Bacillus subtilis,
Cellulases Banana fruit stalk wastes, soyabean meal,
Aspergillus sp
Talaromycesflavus, Citrus wastes, soy bran and wheat bran,
Pectinases
Aspergillus niger apple pectin.
Polymers
Agrobacterium Spent malt grain or ivory nutshaving or
Succinoglycan tumefaciens, Rhizobium grated carrots, impregnated spent malt
hedysaris grains
Spent malt grains, citrus peels, apple
Xanthan gum Xanthomonascampestris pomace, or grape pomace, Impregnated
spent malt grains.

Conditions for SSF

The technical and economic success of SSF is determined by a number of factors,

some of which are listed below:

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i) The choice of substrate: It determines the fermentation by-products and

biomolecules.

Material used for substate: For production of enzymes like amylase, starch-based
substrates are used as material for SSF, while for the production of cellulase enzymes
cellulosic or lignocellulosic substrates are used.The morphology and chemical
composition of the substrate play crucial roles for enzyme production. (Soccol et
al.,2017). Based on the type of substrate under consideration, SSF can be of two types:
SSF with non-reactive (inert) materials acting as mere support and;

Noninert materials, such as biomass which act as support, and serve as carbon and
nutrient sources to promote microbial growth (Carbou et al., 2018)

Particle size of substrate: Substrates with finer particle sizes provide a fixed geometry
and a greater surface area: volume ratio, show better enzyme production than substrates
with larger particle sizes. But if the particle is too small, it causes agglomeration and
interferes with microbial respiration, decreasing the microbial growth (Oriol et al., 1988).

Moisture: Substrate moisture influences the SSF process significantly. Low substrate
moisture is unfavourable for microbes, resulting in their poor growth, while high moisture
content serves as an obstacle to oxygen penetration and hence slow down the process.
Since the substrate is the fundamental parameter for microbial growth,kineticsdepend on
it (Thomas et al., 2013).

ii) The choice of microorganisms:

The choice of the microbes is apparently based on the selection of the substrate and
desired product.Fungi and yeast are the most preferred microbes for SSF, as they thrive
on solid medium and are able penetrate it with their hyphae and rhizoids, and their water
activity is suited for SSF.Bacterial contamination can be avoided in fungal SSF by
increasing the substrate: moisture ratio.The microbiological components of SSF can occur
as single pure cultures, mixed identifiable cultures or totally mixed indigenous
microorganisms. In bacterial species, Bacillus and Clostridium are the potential bacteria,
while Aspergillus, Trichoderma, and Mucor are well-known fungal species for the SSF
process (Sangsurasak et al.,1996). Additionally, filamentous fungi are best suited to
produce industrially important enzymes by solid state fermentation.

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iii) Physio-chemical characteristics:

Volume of inoculum volume, viability, and vegetative cells’ ability are the first
physiochemical parameters that determine the growth of mycelium and microbes and
their interactions with the substrate in medium.

Moisture: The moisture level is another important parameter. A moisture level in range of
60-70% is generally suited for both fungi and bacteria under the SSF condition. The
requirements for water by microorganisms is expressed as the water activity (Aw) of the
microorganisms and not the amount of water present in the solid substrate (Desgranges et
al., 1991). Bacteria mainly grow at higher aw values, while filamentous fungi and some
yeasts can grow at lower aw value (0.6-0.7)

The optimum pH for action of fungal metabolic activity is 4-5. pH: It is a significant
parameter which makes the SSF process efficient. However, the accumulation of organic
acids as fermentation by-products causes a decline in pH, it can be maintained by other
salts present in the medium.

Temperature:In general, SSF is carried outat optimum levels by mesophilic

microorganisms. Particularly, fungi can survive in the wide range of temperature of 20°C-
55°C. The maximum product output is obtained as microbes function best at their
optimum temperature (Penaloza et al., 1991).

Nutrient requirements: The fulfilment of nutritional demands of the microorganism

during the SSF is a strong determinant of the output. Macro- and microelements are a pre-
requisite and improve the metabolic activities of microorganisms during the fermentation
(Krishna et al., 2001). A Carbon/Nitrogen ratio of 16 is best suited for the composition of
the substrate and is used for fermentation processes (Krishna et al., 2005).

SSF Treatment of Crop Residues for Ruminant Feeding

Crop residues are commonly known as ―lignocellulosics‖ because they have high
content of cellulose and are associated with the biopolymer lignin. Even with the assistance
of hydrolytic enzymes, the rumen microbiota (bacteria, protozoa, and fungus) are not capable
of efficiently cleaving these bonds. White rot fungi (WRF) can break the ligno-cellulose

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complexes in such crop residues, releasing free cellulose and thus increasing their feeding
value for ruminants. Compared to untreated roughages, biologically treated roughages exhibit
greater digestibility for most nutrients (both cell walls and cell solubles) and a greater crude
protein content and more fermentable carbohydrates.

Furthermore, recent findings have shown that feedstuffs exposed to solid state
fermentation (SSF) using fungi result in lower methanogenesis as a result of enhanced
digestion and nutrient absorption, and a decrease in structural carbohydrates. Kamra and
Zadrazil (1988) elucidated that when the product is meant for ruminant feeding, the
bioconversion procedure should increase lignocellulose digestibility. The biological
upgradation of crop residues into animal feed should be characterised by considerable lignin
degradation and nutrient liberation from the matrix and the accumulation of digestible
components (Zadrazil et al., 1999) as enhancing the nutritional status of the finished product
using microbial protein. Silva et al. (2002) reduced substrate fibre and CP levels treated with
Pleurotuspulmonaris. However, all fungi do not improve the digestibility of straw. Jalc et al.
(1994) reported that during bioconversion of wheat straw with Polyporus ciliates,
digestibility was improved whereas with Lentinustigrinus, it was reduced.

A major proportion of animal trials on the application of fungal-treated agro wastes

elucidated favourable nutrient utilisation response, nitrogen (N) balance and gain in body
weight (Walli et al., 1988; Mahesh 2012; Shrivastava et al., 2012) although it is not
consistent with all types of fungi. It is an established fact that good quality forage NDF in
lactating cow diet is needed to maintain rumen functioning and optimum milk yield
(Robinson and McQueen, 1992), so fungal treatment of fibrous feed improves digestible NDF
and hence improves milk yield. Ward and Perry (1982) reported an improved digestibility of
DM and NFE of corn cobs treated with Trichodermaviride in lambs. Fazaeli et al. (2004)
observed that inclusion of fungal treated straw upto 30% of the total mixed ration in lactating
Holstein cows improved the digestibility and an increase in fat corrected milk yield by 13%
and average body weight gain. Ruminants release more CH4 on fibrous diets. Mahesh (2012)
observed a linear reduction in CH4 (%) from fungal treated wheat straws which contained
lesser fibre fractions (NDF and ADF) than untreated straw. Abo-Donia et al. (2005) and
Omer et al. (2012) reported a substantial improvement in ruminal pH and the NH3-N
concentration of the rumen liquor in biologically treated groundnut hulls and sugarcane
bagasse, and Trichodermaressi treated corn stalks (Omer et al., 2012).

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Differences between Solid State Fermentation (SSF) and Submerged Liquid

Fermentation (SLF)

Solid State Fermentation (SSF) Submerged Liquid Fermentation (SLF)

Preferred organisms need less water for Water content is much higher than the media
growth. Eg: filamentous fungi. concentration
The inert support (natural or artificial),
The essential processed ingredients are
containing all components for growth are in
expensive.
the form of solution.
Less probability of contamination as there is Higher water activity is the major cause of
lesser availability of water contamination in SLF.
Large-scale bioreactors are required as the
Small size bioreactors can be used.
volume of the media is more
Less consumption of energy is needed for Power consumption is more due to high air
aeration and gas transfer. pressure
The limiting factor for growth is diffusion of
Vigorous mixing makes diffusion easy
nutri
Downstream processing is easier, cheaper and The aqueous form makes downstream process
less time consuming. difficult and expensive
Liquid waste is not produced High quantity of liquid waste is produced
(Manpreet et al., 2005)
Limitations of SSF

 Microorganisms that can only withstand low moisture levels can be employed.
 Precise monitoring of SSF conditions (e.g., O2 and CO2 levels, moisture content) is
not possible.
 As a result of the sluggish growth of the organisms, product creation is limited.
 Heat generation causes issues, and controlling the growing environment is quite
challenging.

Conclusions

SSF is a potential clean technology for producing microbial metabolites from solid
substrates like agro-industrial waste. SSF is a cost-effective green technology for the
production of high-efficiency metabolites. Many value-added compounds are produced using
SSF at the industrial level. To make the process more efficient, this technology must be
improved in the areas of mass transfer, aeration, agitation, and maximal substrate conversion
into products. This field has enormous potential, with the ability to expand to a bigger scale

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in additional sectors in the near future to generate more industrially relevant metabolites and
produce cost effective animal feed.

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