Preface Animal protein sources such as meat, egg and milk are complete proteins as they provide all essential amino acids in highest quality. To provide high quality protein to humans, livestock have to be fed with toxin free quality feedstuffs. Animal feed plays a major role in the global food industry, as it has highest returns to the producer throughout the world. Animal feeds are produced either by livestock farmers themselves locally or feed manufacturing companies. Feeds produced have to be secured and stored in a proper manner to avoid microbial as well as any other biotic agents. Mycotoxins are such naturally occurring toxin produced by the fungal growth. The molds can grow in any of the feed stuffs like maize, other cereals, Groundnut cake and other oil cakes in warm humid climatic conditions. Mycotoxins can have variety of serious health hazards ranging from mild irritation to deadly cancers in livestock and humans. Addressing the issues related to mycotoxin is the need of the hour. With globalisation of the feed industry and rapid climatic changes, it is very difficult to enumerate the occurrence and presence of the toxin in the feedstuff. These unnoticeable toxin effects may have direct and indirect impact on animal production system. It is very essential to have adequate knowledge about the metabolism, growth and reproduction of various mycotoxins and the toxicological properties of their intermediate products produced during metabolism to neutralize these toxins. This book gives a basic idea about the mycotoxins, its various types, conditions suitable for their growth, effect of mycotoxins on rumen fermentation, effect of mycotoxin on animal performance, af latoxin M1 in milk, estimation of mycotoxins, their control measures and application of HACCP in mycotoxin control.

Mycotoxins are the secondary metabolites produced by moulds, which grow on food and feedstuffs causing pathological or physiological changes to humans and animals. They are toxic to vertebrates in low concentrations. Symptoms depend on the amount of toxin in the feed, the period for which the infested feed is ingested, the nutritional status of the feed and the susceptibility of the animal species. Toxic effects of chronic consumption of mycotoxins in human are carcinogenicity, hepatotoxicity, oestrogenicity, genotoxicity, nephrotoxicity, immunosuppression, reproductive disorders and dermal irritation (Anfossi et al., 2010). The most extensively studied mycotoxins viz., Aflatoxins, ochratoxins, deoxynivalenol (DON, vomitoxin), T-2 toxin, zearalenone, fumoninsins, mycophenolic acid, cyclopiazonic acid and ergot toxins are produced by the moulds Aspergillus, Fusarium, Pennicillium and Claviceps (Iqbal et al., 2015).

Various physical, chemical and biological factors interdependently affect the fungal colonization or production of the mycotoxin. Physical factors such as temperature, relative humidity and insect infestations create ideal environment for fungal colonization and mycotoxin production. Stresses such as drought, an increase in temperature, and an increase in relative humidity may selectively alter colonization and metabolism of mycotoxigenic fungi and thus alter mycotoxin production (Russell et al., 1991). Cool and damp weather favours the germination of the sclerotia and thus ergot alkaloid formation in fescue and ryegrass. Biological factors are categorized into intrinsic factor such as fungal species, strain specificity, strain variation and instability of toxigenic properties. Interaction between the toxigenic fungal species and substrate affect the fungal colonization and mycotoxin production. Some plant species are more susceptible to colonization and some resistant plant species may also get infestation due to favourable environmental conditions.

The ingestion of mycotoxin-contaminated feedstuffs by animals leads to adverse effect on animal health and the effects are more serious in monogastric animals depending on the species and the susceptibility to toxins within the species. Ruminants are less susceptible to mycotoxins than monogastric animals because of biotransformation of mycotoxins to non-toxic metabolites in rumen by rumen microbiota (Fink-Gremmels, 2008).

Mould growth in raw materials or finished feeds result in production of mycotoxins. Mycotoxins are a large group of toxic chemicals causing wide range of pathological and physiological effects in animals. Symptoms depend on the amount of toxin present in the feed, the period for which the feed is ingested, the nutritional status of the feed and the susceptibility of the animal. Feed efficiency, growth rate and production are affected in animals consuming sub-acute doses. Ruminants are generally considered less susceptible to mycotoxins than monogastric animals because of degradation of mycotoxin in rumen by rumen microorganism. However, certain mycotoxins resist rumen degradation and cause clinical mycotoxicosis. In high yielding dairy animals, during transition period, animals who are in negative energy balance (NEB), low body condition score and decreased resistance to infectious agents cause adverse effects in animal health with exposure of mycotoxins present in animal diet. Effect of mycotoxin in ruminants In cattle during metabolism, mycotoxin present in stored or ensiled feeds produce microbial volatile organic compounds (MVOCs) which are responsible for mouldy odour, reduced feed intake and increased feeding times. This insufficient feed intake further produces negative energy balance (NEB).

Different type of mycotoxins commonly present in different feed and fodder crops are produced by mould during cultivation and storage and are specific to certain region according to climatic and environmental conditions. Most of the toxins produced in crops are consumed when fed to animals. Among the mycotoxins, aflatoxin B-1 is present in highest concentration which enters rumen following ingestion of aflatoxin contaminated feed stuffs. In the rumen, AFB-1 is degraded into aflatoxicol and the remaining portion of the aflatoxin is absorbed in the digestive tract by passive diffusion and metabolized in the liver into aflatoxin M-1 (Kuilman et al., 1998). AFM-1 circulates in the blood as a stable compound until excreted in milk, urine and bile. The following are the sources of AFM-1 contamination in milk and milk products and it occurs through two modes of contamination viz., indirect contamination and direct contamination.

Mycotoxins are toxic secondary metabolites produced by different types of moulds which are present in soil, plant and feed ingredients. Ingestion of toxins will result in reduced growth and production performance due to reduced nutrient utilization, illness or death of animals. Analysing the presence of mycotoxin and its concentration in grain and feeds is necessary to eliminate the adverse health effects of mycotoxins in animals. Estimation of mycotoxin in feed ingredients is a challenging task because the concentration of toxin is at µg/kg or parts-per billion levels. At the same time, analysing inappropriate samples is a waste of resources. Mycotoxins are not evenly distributed in feeds or grains. If the specimen collected for analysis includes contaminated grain, the mycotoxin will be detected. If not, the mycotoxin will not be detected. Sample collection for mycotoxin analysis The objective of sample collection is to collect the specimen samples from as many parts of the feed lot as possible. The size of the collected specimen depends on the size of the load received. Sample collecting probes can be used to collect the samples from bags.

The economic loss in livestock arising due to mycotoxicosis is enormous apart from being a health risk. Indian economy is heavily affected by mycotoxins. It was estimated that 10 million dollars were lost in India’s export within a decade due to groundnut contamination with mycotoxins (Vasanthi and Bhat, 1998). Many developing countries have realized that reducing mycotoxin contamination in foods will not only improve the health and production in animal, but also increase the raw material exports (international trade) to other countries. The following strategies are to be adhered strictly to reduce the mycotoxins contamination of crops in the field. 1. Early harvesting Early harvesting reduces the fungal infection of crop in the field. Early harvesting and threshing of groundnuts results in lower aflatoxin levels and higher gross returns of 27% than in delayed harvesting (Rachaputi et al., 2002).

Hazard Analysis Critical Control Points (HACCP) is an internationally recognized method of identifying and managing food safety related risk and, when central to an active food safety program, can provide customers, the public and regulatory agencies assurance that a food safety program is well managed. HACCP is a management system in which food safety is addressed through the analysis and control of biological, chemical, and physical hazards from raw material production, procurement and handling, to manufacturing, distribution and consumption of the finished product. (FAO, 2003) Mycotoxin hazard analysis and identification of possible control measures I. Hazard Analysis a) Identification of mycotoxin hazard For a given commodity system in a particular location, the HACCP team need to first consider which, if any, of the mycotoxins known to constitute a food safety hazard are likely to be present. Over 300 mycotoxins are known, but only a relatively few of these are widely accepted as presenting a significant food or animal feed safety risk. The following mycotoxins have regulatory limits set by one or more countries: the aflatoxins (including aflatoxin M1), ochratoxin A, zearalenone, patulin, ergot alkaloids, and deoxynivalenol. Guideline limits exist for fumonisin B1 and regulatory limits are likely to be set in the near future. The regulatory limits are taken as the target levels and should be included in the Product Description table. Mycotoxin limits can also be set by the customer in specific contracts and it is possible that these may include mycotoxins not subject to regulatory limits.

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