Open access peer-reviewed chapter

Benefits of Probiotics on Aflatoxin Infected Birds

Written By

Muhammed Jimoh Ibrahim

Submitted: March 24th, 2021Reviewed: August 3rd, 2021Published: April 6th, 2022

DOI: 10.5772/intechopen.99800

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Abstract

Aflatoxin are transferred from feed to animal products (Eggs, Meats and Milk). There is need to find alternative chemicals that is economically friendly to reduce the impact of aflatoxins. Probiotics additives especially Lactobacillus and Bacillus spp. biodegradation generally decreases aflatoxin residues in milk, egg and meat. They are low cost, economically friendly and accessible additives which could mitigate aflatoxin formation in feed and food. There is need for aggressive public health awareness on the implication of aflatoxin residues and as well as detoxification strategy that can reduce toxin absorption into animal feed.

Keywords

  • Probiotics
  • birds
  • aflatoxin
  • residues
  • implication

1. Introduction

Food safety is effectively achieved when the food pillars, such as; food availability, food access, food utilization, and food stability which permit individual at any time to have access to affordable, safe and healthy food to meet daily nutrient requirement [1]. Weakens of this four pillar pose a treat to food security. Human health and animal welfare are influenced by food insecurity and contaminant, which reflect on social and economic status of a society. Mycotoxin during pre, processing and post-harvest are driving factors of food insecurity since contamination occurs along the food value chain from farm to fork [2]. Poultry products are important international food commodity. Economic losses may occur due to the presence of natural feed contaminants, such as mycotoxins, which are secondary metabolites produced by certain toxigenic aflatoxins [3], poultry-derived products such as meat and eggs are carry-over of aflatoxin into the human food value chain which serve as potential threat to human health [4, 5, 6, 7]. Contaminated food and feeds with aflatoxin prohibit trade of international concern [8]. The regulations on “acceptable health risk” usually depend on a country’s level of economic development, extent of consumption of high-risk crops, and the susceptibility to contamination of crops to be regulated [9]. Safety limit of aflatoxin consumption for human ranges 4–30 mg/kg. European Union has set the strictest standards, which establishes that any product for direct human consumption cannot be marketed with a concentration of AF-B1 and total AFs greater than 2 mg/kg and 4 mg/kg, respectively [10, 11, 12]. Likewise, US regulations have specified the maximum acceptable limit for AFs at 20 mg/kg [13, 14, 15, 16]. Worldwide European Union aflatoxin standard is adopted, meeting this standard Sub-Sahara Africa and Asia encounter both economic losses and financial costs. This situation requires alternative technologies at pre- and post-harvest levels aimed to minimize contamination of commercial foods and feeds, at least to ensure that AF levels remain below safe limits [15, 16].

Physical, chemical and biological approaches have been conducted to degrade mycotoxin. Most of these method are unsafe due to losses in the nutritional value, cost of equipment, and formation of intermediate metabolite [17]. Biological detoxification using microorganisms or enzymatic preparations is promising [18]. Probioitcs such as Rhodococcus erythropolis, Armillariella tabescens, and Myxococcus fulvu, Rhizopus oryzaes, Pseudomonas spand Bacillus subtilis, have been reported to have different AF-degrading ability [19, 20, 21]. Bacillus subtilisapplied directly on the feedstuffs degrade 81.5% AFB1 and 85% ZEA in naturally contaminated feed in vitro [22, 23]. B. subtilishad protective effects against aflatoxicosis in layers and broilers fed naturally AF-contaminated diets [24, 25, 26]. It is therefore, important to identify benefits of probiotics on aflatoxin contaminated poultry products to effectively monitor carry-over of residues to sustain healthy living and socioeconomic development.

1.1 Mycotoxin

Mycotoxin refers to harmful secondary metabolites produced by fungi in food and feed products that negatively impact animal and human health, by themselves or through synergistic interactions with each other [27]. Mycotoxins are structurally diverse low-molecular weight secondary metabolites produced by fungal growth [27]. Aspergillus, Penicillium, and Fusarium contaminate feed and food consumed by animals and humans. Globally, millions of dollars are losses annually on mycotoxins, on agricultural products, animal and human health [15].

1.2 Aflatoxins

Aflatoxins are polyketide secondary metabolites produced by toxigenic strains of Aspergillus, Penicillium, Fusariumand Alternariafungi [28, 29]. They grow on a variety of nutritional substrates like cereals which is the main active ingredient of poultry and human food [30]. They are extremely harmful to the health of humans and animals, showing changes in biochemical and hematological indices effecting metabolism via alteration of enzymatic pathways of starch, proteins, lipids and nucleic acids. Hence, serum glutamate pyruvatate transaminase, serum gluatamate oxaloacetate tranferase and γ-glutamyl transferase activities are increased, inciting; hepatotoxic, carcinogenic, mutagenic, teratogenic, immunosuppressive actions and in severe intoxications may cause death [31, 32, 33, 34, 35, 36, 37, 38]. Acute or chronic aflatoxicosis in poultry results in retarded growth, decreased production and egg quality, impaired immune response, increased mortality and liver and intestine damage [39, 40]. AF is also known to interfere with metabolism of vitamin D, iron and copper and can cause leg weakness. Aflatoxin has caused serious destructions in Africa, which has caused significant financial losses in agricultural commodities contaminated with toxins and consequently having effects on animal and human health point of view [41, 42]. Although most countries of the world has been affected by aflatoxin, it is sub-saharan Africa (SSA) that has suffered most [43]. Most of SSA agriculture occurs in impoverished rural areas and a lack of technical infrastructure in many African countries does not allow for routine quality control of even commercially produced commodities, never mind those produced by rural population for their own consumption [43]. Ultimately, the transmission of AF and its metabolites from feed to animal edible tissues and products, such as liver and eggs, becomes a potential hazard for human health.

1.2.1 Types of aflatoxins

Among the 18 different types of aflatoxins identified, the major members are aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2), which are produced by Aspergillus flavusand Aspergillus parasiticus A. nomius[44]. M1 (AFM1) and M2 (AFM2) are metabolites of AFB1 and AFB2 in human and animal milk fed on contaminated food. Aflatoxin B1 (AFB1) being the most toxic among other species. Additionally other species which produce aflatoxin are A. pseudotamarii, A. ochraceoroseus, A. rambellii, A. toxicarius[45]. In addition other fungi of the genera Aspergillus(e.g. A. ochraceusand A. carbonarius) produces another important mycotoxin ochratoxin A (OTA) [38, 46]. A. flavusand A. parasiticusvaries from highly toxigenic to non-toxigenic forms and are produced by AFB1 than AFG1. A. parasiticusare produce by AFB1 and varying amounts of AFB2, AFG1 and AFG2 with variable toxigenicity [47]. Aflatoxins B occur more frequently as contaminants, and are also believed to be more potent, than Aflatoxins G [48].

1.2.2 Chemical structure

Chemically aflatoxin B occur they are difuro-coumorins –cyclopentenone and difurocoumaro lactone series which are freely soluble in chloroform and methanol [49, 50], Other aflatoxins have different substitutes but share basic coumarine structure. The epoxidation of the 8, 9-double bond and cyclopentenone ring of B series is responsible for the order of acute and chronic toxicity as compared with the six-membered lactone ring of the G series AFB1 > AFG1 > AFB2 > AFG2 (Figure 1) [49].

Figure 1.

Chemical properties of aflatoxin B and G (A–F). Source: Adapted from Agriopoulou et al. [38].

1.2.3 Physical structure

Structurally they are dihydrofuran-coumorins moiety containing double bond which are freely soluble in chloroform and methanol. They are stable at high temperatures but unstable to UV light or polar solvents [49, 51]. Aflatoxins are toxic secondary metabolites upon exposure to fluorescence ultra violet (UV) light, aflatoxin B appear blue in color and G appear green in color (Table 1) [49, 52].

AflatoxinMolecular formularMolecular weightMelting point °C
B1C17H12O6312268–269
B2C17H14O6314286–289
G1C17H12O7328244–246
G2C17H14O7330237–240
M1C17H12O7328299
M2C17H14O7330293
B2AC17H14O7330240
G2AC17H14O8346190

Table 1.

Physical properties of aflatoxins.

Source: International Crop Research Institute for Semi-Arid Tropics.

Adapted from: Reddy et al. [49]

1.3 Occurrence of aflatoxin in food and feed

Eggs, milk and meat are sometimes contain residues of aflatoxins because of consumption of aflatoxin contaminated feed ingredients such as peanuts, cottonseed, nuts, almonds, figs, spices, soybean, rice and maize [53].

1.4 Mode of action

Cytochrome P450 enzymes (phase I metabolisation) convert aflatoxins to a reactive 8,9-epoxide form, which is essential for the toxicity. In mammals CYP1A2 and CYP3A4 are the enzymes responsible for conversion [54] in chicken and turkeys, the corresponding enzymes are CYP2A6 and to a lesser extent CYP1A1 orthologs [55, 56]. DNA and protein binds to guanine residues of nucleic acids to produced epoxide metabolite causing genotoxicity and cytotoxicity [57]. Aflatoxin B1-DNA adducts result in guanine-cytosine (GC) to thymine-adenine (TA) transversions [48], which leads to irreversible DNA damage, therefore results to hepatocellular carcinomas [58]. Gluthatione conjugation or hydrolysis detoxified the toxic epoxide metabolite and epoxide hydrolase to phase II metabolisation and AFB1–8,9-dihydrodiol (AFB1-dhd) respectively. AFBI Metabolisation to less toxic compounds such as aflatoxin M1 (AFM1) or Q1 (AFQ1) [54, 56]. AFM1 metabolite possesses carcinogenic properties which are 10 times lower than AFB1. These metabolites obtained from cattle milk. The maximum limits in milk permissible for human consumption have been established (0.05 μg/kg) [12, 59], 20 ppb in grain and 4 ppb in food and agricultural commodities [59].

1.4.1 Carcinogenesis

The International Agency for Research on Cancer [60] classify aflatoxin as class 1 carcinogen, transversion of G to T occur in guanine codon 249 of tumor suppressor gene p53 of DNA that induce mutagenesis by alkylation of nuclear DNA, leading to carcinogenesis and teratogenesis [61]. 8, 9,-epoxide is a potent carcinogen and induces chromosomal aberrations, mutation and cell toxicity [62].

1.4.2 Immunesuppression

Immunosuppressive effects on NK cell activity, humoral and cellular immune function are impair by aflatoxin through reducing the primary and secondary immune responses [63, 64, 65, 66]. AFB1 induces; thymic aplasia, reduce T-lymphocyte function, lymphokines, suppress phagocytic and complement activity [67, 68]. Aflatoxin suppresses the levels of IL-1, IL-2, IL-6, IFN, TNF alpha, mRNA and proinflammatory cytokines [69, 70]. Embryonic chicks exposed to AFB1 showed a depressed graft-versus-host response, thymic bursal involution, delayed cutaneous hypersensitivity, macrophages function, reduced antibody titers to vaccines for Newcastle, Mareks and infectious bursal disease [32, 52, 71, 72].

1.4.3 Nutritional

In poultry a drop in feed conversion efficiency and decreased growth rate is observed following a chronic exposure to aflatoxin feed [73]. Aflatoxin modifies vitamin A nutrition in poultry halving the serum retinol and Plasma concentration of 25-hydroxyvitamin D and 1,25- dihydroxyvitamin D concentrations [48, 74]. Bennett and Klich [8], toxin has been a factor modulating the rate of recovery from protein malnutrition. Toxin contaminated diet affect zinc and selenium which are essential for healthy immune systems [75].

1.4.4 Aflatoxin control

Contamination of feed and food with aflatoxins occur during the preparation value chain. Several methods have been adopted in the prevention of aflatoxicosis in animal origin. Application of Good Agricultural Practices (GAP) are important strategy during pre-harvest. Appropriate GAP includes crop rotation, soil cultivation, irrigation and proper use of chemicals. Crop rotation is important and focuses on breaking the chain of infectious material, for example by maize/legume rotations. Any crop husbandry that includes destruction, removal or burial of the infected crop is seen as good soil cultivation. The deeper the soil is inverted (plowing), [76]. Reducing plant stress by irrigation is also valuable to prevent fungi infestation [77]. Damages caused by insects, birds and rodents increases susceptibility of aflatoxin invasion. Successive fungal infection must by controlled by appropriate use of critical pest management system and application of fungicides [77]. Climate change such as high temperature, relative humidity and drought influenced mold infection and mycotoxin production [17].

Mycotoxin are prevented during storage by improving the post-harvest storage conditions [78]. Jard et al. [79], reported storage of grain at less than 15% moisture, removal of infected grain by insect and visibly damaged this prevent favorable condition for mold growth, combination of multiple strategies to reduced moisture content of grain and prevent mold formations. Mycotoxin are destroyed, inactivated, or generate non-toxic products which do not altered the nutritional quality of the food or feed [79]. There are several decontamination processes which include radiation, oxidation, reduction, ammonization, alkalization, acidification and deamination [17]. These chemical methods are not allowed in the European Union [12] as chemical transformation might lead to toxic derivatives. In the United States, only ammonization is licensed for detoxifying aflatoxins.

1.4.5 Detoxifying

Detoxification of agricultural commodities through; radiation, oxidation, reduction, ammonization, alkalization, acidification and deamination is restricted due to problems associated with incomplete detoxification, cost implication and unavailability of equipment. Commonly used method to reduce mycotoxin exposure in the field is the inclusion of mycotoxin detoxifying agent in feed (mycotoxin detoxifiers) which decreases the bioavailability of the toxin [79, 80]. There are two different class of detoxifiers, namely mycotoxin binders and mycotoxin modifiers. The modes of action differs; mycotoxin binders adsorb the toxin in the gut, resulting in the excretion of toxin-binder complex in the feces, whereas mycotoxin modifiers transform the toxin into non-toxic metabolites [34]. Detoxifier are extensively use as feed additives for the reduction of contamination of feed by mycotoxin; which modify their mode of action, reduce absorption and secretion of metabolites [34]. Detoxifier does not mean that animal feed exceeding maximal regulatory limits used. Quality of feed can be improve by adding detoxifier making the product acceptable in market and providing safety for animal health [80].

1.4.6 Organic binder

Lactic acid bacteria (LAB), are divided into four genera: Lactococcus, Lactobacillus, Leuconostoc and Pediococcus. They are Gram-positive, catalase-negative, non-sporulating, usually non-motile rods, cocci, ferment carbohydrates, produced lactic acid [81]. Lactic acid bacteria are used in food processing industry for fermentation, preservation and mycotoxin binding abilities [82]. The mechanism of interaction involves the peptidoglycan structure (amino acid) which are common site for binding. However, different mycotoxin have different binding sites [82].

1.4.7 Probiotics

Application of biotechnological tools to reduced chemical residues and improved production efficiency that does not create any harm to poultry as well as consumers of the value chain [83]. Recent advancement in biotechnology on poultry feeds, banning of harmful growth promoters and antibiotics. Globally, probiotics is gaining acceptance in feed formulation [83]. Antimicrobial resistance is now a worldwide threat [84] with alteration of immune response due to feeding of antibiotic growth promoters, Probiotics are considered as an important tool as regard to antimicrobial resistance [85]. Chick gut are usually sterile immediately after hatch, colonization of microflora on the gut occur on the hatching tray, hatcher, feed and water intake. These Microorganisms in the gut could either be beneficial or harmful based on their response to the host immune system. The beneficial organisms maintain gut equilibrium, improve health and production of the birds. However, harmful bacteria like E. coli, Salmonella, Coliformand Campylobacteradjust the gut equilibrium to favor spread of infection. Probiotics supplementation mitigate the spread of infection on poultry. Commercial probiotics preparation can be administer as a single or multi-strain where they positively improved production and egg shell quality [86]. Probiotics depends on several factors for their survival on the host, this include; dose frequency, type of host animal, strain and stability of organism, genetic component of host, nutritional status of host age and physiological levels [87, 88]. Research findings showed that use of probiotics in layer diets enhanced egg production, improve body weight [89, 90, 91, 92], reduced serum low density lipoprotein (LDL) cholesterol [93], decrease cholesterol and triglycerides in blood [94, 95]. Probiotics improved shell quality hardness and bone strength in laying hens [96]. Improvement in the production of darker yolk color Sobczak and Kozłowski [90].

1.4.8 Lactobacillus spp.and Bacillus spp.

Physical and chemical detoxification are associated with some disadvantages such as undesirable effects on products, loss of nutritional quality and altered organoleptic properties, high cost of production and time consumption [97]. Antibiotic are used in poultry to treat an infection, growth promoter and productivity thus causes antimicrobial resistance to the health of livestock and consumers of the bye products [98]. Multi-drug resistance genes (MDRG) occurs due to under administration, overdose, drug residues and extra label use of drugs which is emerging in both animal and human due to continuous use of antibiotic in the diet of poultry. However, biological methods based on competitive exclusion where probiotics colonized adhesive sites on the intestinal epithelium thereby, prevent colony formation of pathogenic bacteria, non-toxigenic fungal strains have been reported promising method for lessening the formation of mycotoxins and preventing their absorption animal to human [87, 99]. Lactobacillus, Bifidobacterium, Propionibacterium, and Lactococcus are found to be active in terms of binding AF-B1 and AF-M1 [97, 100, 101]. Probiotics are alternatives for growth promotion, food safety, enhanced nutrient assimilation, improve production and reducing harmful bacterial concentration of the gut [87, 102, 103]. Binding of aflatoxin depend on several factors such as temperature, incubation time, pH, matrix and strain of probiotics [104]. Probiotics act as antagonist against aflatoxin, by altering metabolism of gastrointestinal tract, production of volatile fatty acid, organic acid, antibacterial (lactocidin, acidophillin, bacteriocins and hydrogen peroxide), stimulation of essential nutrient for immune responses and inhibiting bacteria growth [105, 106]. Absorption of nutrient and digestive activity are increase with decreased in ammonia production and bacteria enzyme activity (glucoronidase, nitroreductase, azoreductase) produced by pathogenic bacteria. They stimulate immune system by higher production of immunoglobulins, macrophages, lymphocytes, γ-interferon increase villus height, goblet cells and crypt depth to create environment unfavorable to agent [107]. Strain composition and doses determines the potentiality of probiotics [86]. Single strain probiotics exact direct mechanism of action but for multi-strain, it exact synergistic synergistic action among different strains and in such condition, it is supposed that multi-strain probiotics have more adhesive power than single strain [108].

1.4.9 Intestine

Intestine and the intestinal epithelial cell layer are selective barrier between external and internal environment. The first barrier layer prevent exposure of high concentration of foreign antigens, natural toxins, pathogens and mycotoxin [109, 110]. Intestine are maintained by well-organized intercellular structures including tight junctions, adherence junctions and desmosomes surrounding the apical region of epithelial cells [111]. Physical and chemical factors can dynamically alter the structure and function of tight junctions. The trans-epithelial electrical resistance (TEER) of cell monolayers can be considered as a good indicator of the epithelial integrity and of the degree of organization of the tight junctions over the cell monolayer [112]. The primary function of intestinal cells are to act as a physical barrier, separating the contents of a harsh luminal environment from the layers of tissue comprising the internal milieu [113]. Intestinal epithelial cell studies performed on rats indicate that aflatoxin B1 decreases intestinal cell proliferation throughout the intestine [114]. The intestinal epithelial cells barrier function as both on innate and adaptive components of immunity [113].

1.5 Immune response

Various mycotoxins affect immune-related organs and cells, and influence host defenses against infectious agents and related microbial toxins [115]. Aflatoxins suppress immune functions, particularly cell-mediated immune responses [116]. For instance, high levels of aflatoxin B1 (AFB1)-albumin adducts change T-cell phenotypes and reduce the percentage of B cells in human immunodeficiency virus-positive individuals [117]. In addition to lymphocytes, embryonic exposure to AFB1 impairs the functions of phagocytes such as macrophages and neutrophils, via the depression of phagocytic potential, inhibition of antiviral activity, and reduction in chemotactic responses [118, 119, 120]. AFB1 also interferes with the innate immunity of macrophages by suppressing tumor necrosis factor-α (TNF- α), interleukin (IL)-1, and IL-6, resulting in the disruption of pulmonary and systemic host defenses [67, 121].

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2. Conclusion

Probiotics significantly counteract the adverse effect of aflatoxins which effectively reduced accumulation of aflatoxin residues in milk, meat and eggs [122]. In conclusion, feed and food industry could benefit from the use of probiotics to mitigate aflatoxin residues in eggs, milk and meats. Hence, probiotics might be promising tools in decreasing economic and health damage caused by aflatoxin in poultry industry. The prevalence of aflatoxin residues in poultry products call for public health attention of food safety along the value chain, by creating awareness on the presence of aflatoxins on poultry products and health implication to both animal and human.

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Written By

Muhammed Jimoh Ibrahim

Submitted: March 24th, 2021Reviewed: August 3rd, 2021Published: April 6th, 2022