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MICROBIOLOGICAL EVALUATION OF PROCESSED AND PRESERVED FEED

Article · June 2020

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Adya Prakash Rath Sudhanya Nath

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MICROBIOLOGICAL EVALUATION OF PROCESSED AND PRESERVED FEED

Dr. Sudhanya Nath1*, Dr. Adya Prakash Rath2, Dr. Saurabh Karunamay3, Dr. Smita Patil4 and
Dr. Diptanu Das5
1
PhD Scholar, Department of Animal Nutrition, West Bengal University of Animal & Fishery
Sciences, Kolkata, West Bengal – 700037, India
2
PhD Scholar, Department of Veterinary Pathology, Guru Angad Dev Veterinary and Animal
Sciences University, Ludhiana, Punjab – 141001, India
3
PhD Scholar, Department of Livestock Products Technology, West Bengal University of
Animal & Fishery Sciences, Kolkata, West Bengal – 700037, India
4
PhD Scholar, Department of Livestock Production Management, West Bengal University of
Animal & Fishery Sciences, Kolkata, West Bengal – 700037, India
5
PG Scholar, Department of Animal Nutrition, College of Veterinary Sciences and Animal
Husbandry, Selesih, Aizawl, Mizoram - 796014

*Correspondence:sudhanyanath@yahoo.com

INTRODUCTION:

Animal feeds are blended or processed products of plant and animal origin whose
principal purpose is to meet animal’s nutritional needs. A number of feed and feed
ingredients are imported and feeds are often produced as processed finished or compound
feeds.
In recent years, there has been a diversification in feed ingredients available and
feeding practices. For example, there has been an increased use of compound feeds in the
dairy industry, reflecting the rapid growth and intensification in that sector. In addition, there
is a growing range of imported feed and feed ingredients from a variety of overseas sources,
which poses an additional risk for the introduction of pathogens and contaminates into the
food chain. There has been interest in further elucidating the relationship between
pathogenic/spoilage bacterial contamination of animal feed and human foodborne illness.
A wide range of microbes occurs naturally on, or as contaminants of forages, cereal
grains, oilseed by-products and compound feeds. Animal feeds may become contaminated
with harmful bacteria such as Salmonella, Listeria and E. coli. In the case of the latter
organism, faecal sources and slurry have been identified as the primary routes of
contamination both in pastures and compound feeds. Cereal grains and oilseed by-products
are regularly contaminated with fungi occurring as plant pathogens or developing during
storage. Major adverse effects arise in farm animals due to the production of mycotoxins by
certain species and strains of these fungi.
Beneficial effects of microbes in feed can result from the occurrence of the lactic acid
bacteria during the fermentation of forages in the process of ensilage. These bacteria favour
the production of lactic acid, which helps in reducing the pH to around 4.0, thus preserving
the forage for winter-feeding to ruminants.
Animal feed, due to its composition, provides a favourable environment for the
growth of microorganisms. Microorganisms found in the feedstuffs can be saprophytic,
pathogenic, conditionally pathogenic and toxic. Their growth and proliferation in the feed

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depends on numerous factors, such as moisture, temperature, type of feed, aerobic and
anaerobic conditions, chemical and physical properties of raw material, feed pH value,
presence of feed supplements, storage periods and conditions as well as feed decomposition
products.
Feed may be contaminated during processing, storage or transport. Some
microorganisms introduced during storage, primarily moulds, can negatively affect feed
quality including reducing dry matter and nutrients, causing musty or sour odours, and
producing toxins. Mouldy raw materials are not appetizing and can considerably reduce feed
consumption. Contaminated feed frequently causes zoonoses and for that reason, it is
necessary to establish surveillance programs for microbiological feed hazards.
The importance of microbial quality of the feed is not being considered in animal
feeding till they affect the life or performance of the host animal. These microbes not only
affect the animals but also the human with respect to salmonellosis, scrapie, listeriosis. The
contamination with fungus leads to not only change in colour, flavor and palatability, but also
causes hepatic and kidney damages due to production of mycotoxins. These toxins also
possess tumour causing properties. The presence of microbial population in silage is very
much important for the preparation of good quality silages. However, this area of animal
nutrition is rarely under studied except the mycotoxins. The importance of microbiological
quality is gaining importance over the past two decades since the report of bovine spongiform
encephalopathy in Europe.
The microbiology of animal feeds emerged as an important issue in the wake of the
Salmonella, E. coli, Campylobacter and Bovine Spongiform Encephalopathy(BSE) crises in
the EU and elsewhere and the foot and mouth epidemic of 2001-02 in United Kingdom.

FUNGAL CONTAMINATION OF CONCENTRATES AND FORAGES

Contamination may occur during processing and storage of harvested products and
feed whenever environmental conditions are appropriate for spoilage fungi. Moisture content
and ambient temperature are key factors affecting fungal colonization of and mycotoxin
production in concentrates and compound feeds. The risks arise primarily from the ability of
particular species and strains of fungi to produce harmful compounds known as mycotoxins.
Fungal contamination of animal feeds is a regular occurrence on a worldwide scale
and detrimental effects have been observed in all classes of farm animals due to the
production of mycotoxins by certain species and strains of moulds. There are few grounds for
complacency as regards animal feed safety and vigilance should now be the watchword for
all those involved in the livestock industry.

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 Toxigenic fungi of concentrates and forages

Fungi Occurrence Mycotoxins

Aspergillus flavus; A.parasiticus Peanut meal, Aflatoxins

cottonseed cake,
palm kernel cake,
maize,compound feeds
A. flavus Oilseed meals, compound Cyclopiazonic acid
Feeds
A. ochraceus; Barley and wheat grains Ochratoxin A
Penicilliumviridicatum; P.
cyclopium
P. citrinum; P. expansum Cereal grains Citrinin

P. citreo-viride Cereal grains Citreoviridin

Fusarium culmorum; Cereal grains Deoxynivalenol

F.graminearum

F. sporotrichioides; F.poae Cereal grains T-2 toxin

F. sporotrichioides; Cereal grains Diacetoxyscirpenol

F.graminearum; F.poae
F. culmorum; F. graminearum; Cereal grains Zearalenone
F. sporotrichioides
F. moniliforme Maize kernels Fumonisins;moniliformin;
fusaric acid

The Aspergillus genus dominates all other fungi in respect of mycotoxin production in
cereals and oilseeds. Some researchers observed that Aspergillus was the most significant
genus in dairy and other feeds in the tropics. Three species are responsible for virtually all
mycotoxin production by this genus: Aspergillus flavus, Aspergillus parasiticus and
Aspergillus ochraceus. A. flavus, and A. parasiticus synthesise the aflatoxins, while A.
ochraceus produces the ochratoxins.
The aflatoxins include aflatoxin B1, B2, G1 and G2 (AFB1, AFB2, AFG1 and AFG2,
respectively). In addition, aflatoxin M1 (AFM1) may well occur in the milk of dairy cows
consuming AFB1-contaminated feeds. The aflatoxigenic Aspergilli are generally regarded as
storage fungi, proliferating under conditions of relatively high moisture / humidity and
temperature.

Salmonella species

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 Many species of Salmonella have been implicated in diseases of farm animals. Of
these, S.typhimurium is universally distributed while S. enteriditis has emerged as a regular
pathogen ofpoultry and contaminant of eggs and chicken meat. Animal feeds are thought to
be an importantsource of these bacteria. Meat and bone meal and fish meal are frequently
contaminated with Salmonella. Intensive pasture utilisation provides an additional source
through contamination offaeces from infected animals. Furthermore, the practice of spreading
cattle slurry on to pasturesin conventional and organic farms is another potentially significant
source of infection.

Escherichia coli
It is widely recognised that cattle feeds contain E. coli through contamination with
faeces. There is particular concern over the occurrence of E. coli O157 since this form has
been definitively linked with specific outbreaks of illness in humans.
The application of slurry on to pastures means that there is potential for the transfer of
faecal E.coli to grazing animals, a practice that has caused some disquiet among those
concerned withfood safety. It was suggested that attention should focus on the replication of
E. coli in moist feeds and duration of storage in feed bunks.

Foot and mouth disease:

Foot and mouth disease has been a major problem in the UK during 2001-02, with
devastating effects on the livestock industry. The disease is spread by inhalation and by
ingestion of contaminated materials. The outbreak in the UK has been tentatively attributed to
the feeding of catering waste to pigs. While this association may never be confirmed, it is
clear that in the intensive rearing of animals due recognition should be given to the need to
correctly process animal feeds. However, in the EU, the feeding of catering waste containing
meat products is banned under a recent directive.

Bovine Spongiform Encephalopathy(BSE)

The prion proteins of processed animal proteins have recently emerged as important
feed contaminants implicated in the development of BSE in cattle. Prion proteins are normal
animal tissue components with the capacity to transform into agents causing fatal
neurological syndromes in a wide range of species. The initial onset of BSE was attributed to
the feeding of cattle with meat-and-bone meal prepared from carcasses of scrapie-infected
sheep. The latter disease is also caused by prion proteins as is the human equivalent, new
variant Creutzfeldt-Jakob Disease (vCJD). The incidence of vCJD in humans has been linked
with the consumption of BSE-contaminated beef. It is this association that has led to
extensive and stringent legislation in the European Union concerning the use of specified
animal products in livestock feeding.

SILAGE MICROBIOLOGY

The process of silage making plays an important role in feeding of green forages
during post-monsoon seasons throughout the continents. Successful preservation of high-

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moisture forage and other crops depends upon the controlling the activities of microbes,
particularly bacteria. Conditions favouring optimal preservation include the rapid imposition
and subsequent maintenance of anaerobic conditions at all stages in the process of ensilage.
Under these conditions, lactic acid bacteria proliferate, using endogenous plant sugars to
produce sufficient quantities of the lactic acid to depress pH to around 4, the optimum value
for successful preservation. Wet forages are difficult to preserve in this way and often
provide ideal conditions for the growth of undesirable bacteria such as the Clostridia.
During ensilage either one of the two types of fermentation (homo- and hetero-
fermentation) takes place and ultimately determine the silage quality. Among these two
fermentation types, the homo-fermentative is more desirable as maximum acidity could be
obtained which inhibits the growth of aerobic organisms, mould and yeast and protect the
silage form spoilage. The maintenance of anaerobic environment during silage making not
only favours the growth of lactic acid bacteria but also the growth of obligate anaerobe
clostridial organisms. The lactic acid bacteria (LAB) are of two types. The homofermentative
species include Lactobacillus plantarum, Pediococcus pentosaceus and Enterococcus
faecalis. The heterofermentative species comprise Lactobacillus brevis and Leuconostoc
mesenteroides. The homofermentative group are more efficient in converting forage sugars
into lactic acid .
Two groups of Clostridia are also recognised. The saccharolytic group includes
Clostridium butyricum and C. tyrobutyricum. These species ferment residual sugars as well
as lactic acid to butyric acid, causing a rise in pH. The proteolytic group includes C.
bifermentans and C. sporogenes. These bacteria ferment amino acids to amines and ammonia
generation which causes ammonia odour as well as increase in the pH toward alkaline side
(>5). This raise in pH accompanied with high moisture facilitates the growth of fungus which
changes the smell, taste and colour which in turn affect the acceptability by the animals. The
production of butyric acid as a result of clostridial fermentation caused reduced feed intake in
ruminants.
The Enterobacteria include Escherichia coli and Erwinia herbicola and are also
considered to be undesirable in that they compete with LAB for plant sugars fermenting them
to acetic acid, ethanol, CO2 and H2. They are also capable of catabolising amino acids to
NH3.
Listeria monocytogenes is widely distributed in nature and also occurs in silage,
particularly big bale silage. The increased incidence of listeriosis in sheep and cattle has been
partly linked with the introduction of big bale silage. The relatively low density and limited
fermentation characteristics of big bale silage and susceptibility of the bags to damage all
favour the growth of L. monocytogenes. This organism is of particular significance because
of it potential to contaminate animal products destined for human consumption.
Fungi occur in silages as yeasts and moulds. The yeasts include species of Candida,
Saccharomyces and Torulopsis. The moulds associated with silages include various species
of Aspergillus, Penicillium and Fusarium. The occurrence of these fungi is of particular
concern due to their potential to produce harmful mycotoxins.
Wilting the crop before ensiling is a common method of restricting fermentation,
allowing the growth of LAB but inhibiting the activities of undesirable organisms such as
Clostridia and Enterobacteria.

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 Microbiology of ensilage
Organisms Conditions required Major products/effects

Lactic acid bacteria Anaerobic; wilting of crop is Homofermentative pathway: lactic

(LAB) desirable; crop should be acid and some acetic acid.
chopped for rapid Heterofermentative pathway:
establishment of LAB. lactic acid, ethanol, mannitol,
acetic acid and CO2.
Clostridia Anaerobic; wet forage Saccharolytic species: butyric
acid, CO2 and H2.
Proteolytic species: butyric acid,
acetic acid, amines, CO2 and
NH3.

Enterobacteria Anaerobic; optimum pH 7.0; Acetic acid, ethanol, CO2 H2 and

active in early stages of NH3.
fermentation.

Listeria Aerobic; pH above 5.5; growth Listeriosis, especially in sheep.

possible at low temperatures
and in high-dry matter silages

Fungi Aerobic; active on surface Spores and mycotoxins.

layers of silage.

MICROBIAL ASSESSMENT :

The assessment of microbial contamination in animal feeds needs to be rapid,

sensitive and representation of diverse population.
The approaches includecollection of samples, isolation of microorganisms bythe
complete enumeration by total plate count (both aerobic and anaerobic organisms),
identification of bacterial and fungal isolates;bacterial isolates were identified based on their
gram staining and biochemical characteristics (indole test, sugar fermentation, methyl red
test, voges- proskauer test, catalase test, coagulase test, citrate test etc.), fungal isolates were
identified based on their morphological characteristics on SDA(Sabouraud Dextrose Agar)
and Lactophenol cotton blue stain identification.Thin layer chromatography (TLC)
quantifies mycotoxins formed and indirectly measure the extent of fungal contamination
during the post-harvest and storage periods.

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 These conventional procedures are less sensitive and time consuming; hence, the
modern techniques of polymerized chain reaction could be used in assessment of feed
microbial quality. This test will be rapid, more sensitive and also representation of diverse
microbial contamination in the feed.

CONCLUSION:

Microbiological safety and cleanness of animal feeds is essential not only due to the
sanitary conditions of animals, but also because of indirect impact on health of consumers.
Therefore, it is important that the producers of animal feeds, having in mind health
consequences in animals and humans who consume food products of animal origin, provide
the highest possible microbiological cleanness of their products. An essential role is also
played by the farmers, who are producers of plant resources – the basic animal feeds for
animals. Both farmers and producers within the framework of hygiene and good practice
must pay attention particularly to the stage of drying and storing feed resources, in order to
prevent microbiological contamination. With continuous collection and evaluation of
microbial safety-related data during processing, distribution and use of animal feed, and
application of adequate agricultural and management practices, microbial feed safety hazards
may be considerably reduced and adequate feed quality ensured. Feed quality and safety are
important prerequisites for sustainable development of livestock production.

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