ASSESSMENT OF AFLATOXINS IN FEEDS AND FEED INGREDIENTS OF BOTH LIVESTOCK AND POULTRY

Assiut University web-site: www.aun.edu.eg

ASSESSMENT OF AFLATOXINS IN FEEDS AND FEED INGREDIENTS OF BOTH LIVESTOCK AND POULTRY

AHMED A. SHARKAWY EL-SHERIF and MOHAMMED A.M. ALI

Forensic Medicine and Toxicology Dept.- Fac. of Vet. Medicine - Assiut Univ. - Egypt.

Received: 30 September 2018;    Accepted: 30 October 2018

INTRODUCTION

Aflatoxins (AFs) are one of mycotoxins produced by many different types of fungi such as asperigillus (Aspergillus parasiticus, Aspergillus flavus, and Aspergillus nominus). Aflatoxins are secondary   metabolites   of   these     fungi     which

Corresponding author: Dr. Ahmed A. Sharkawy El-Sherif

E-mail address: ahmedsharkawy61@yahoo.com

Present address: Forensic Medicine and Toxicology Dept.- Fac. of Vet. Medicine - Assiut Univ. - Egypt.

contaminate a large number of feedstuffs (especially cottonseed, corn, maize, grains, cereals and peanuts) and food. Aflatoxins are difuranocoumarin chemical compounds. According to the chemical structure, there are many types of aflatoxins like B1, B2, G1, G2, M1 and M2. The main aflatoxins types found in milk are M1 and M2. AFM1 as a metabolite of AFB1 and AFB2 can found in eggs. Aflatoxins cause many toxic effects for plants, animals, human as well as microorganisms (Talebi et al., 2011, Chen et al., 2013, Wacco et al., 2014). The production of AFs occurs in field due to the effect high temperature, stress, and drought and (Villers, 2014).

Chemical structures of aflatoxin B1, B2, G1, G2, M1 and M2 (Talebi et al., 2011).

Many of world organizations as FAO found that myctoxins contaminate more than 25% of the agricultural commodities and as a result for this contamination many economic losses were occur (Kabak et al., 2006). Mycotoxins are responsible for many different toxic effects for both animals and human and considered as the most dangerous problem for feeds and food (Sforza et al., 2006). Over 400 well known mycotoxins, aflatoxins are the most famous one as they toxic to human beings (they have high toxicity like hepatotoxicity, carcinogenicity, teratogenicity as well as mutagenicity effects). Patients who already infected with hepatitis B and Hepatitis C are in a high risk for induction of hepatic cancer when consuming foods contaminated with aflatoxins. International Agency for Research on Cancer classified AFB1 as a human carcinogen group 1 (IARC, 2000, Costanzo et al., 2015).

The fungi which responsible for production of aflatoxins utilize the nutrient materials necessary for their growth from the feed and food leading to minimizing the nutritional quality of these types of foods (Akande et al., 2006). In general, birds exposed to feeds contaminated with aflatoxins will suffer from exhausted immune system which reflected in high affections with large number of diseases (Dhanasekaran et al., 2009). Cereal grains, cotton seed, peanut and protein rich meals such as cotton seed meal, corn gluten meal, copra meal, palm kernel meal, rapeseed meal, soybean meal, and sunflower meal considered as the major suitable media for growth and propagation of Aspergillus flavus (Anjum et al., 2012).

Nowadays poultry industry (broilers, duckling, layers, quails and turkeys) is suffering from severe economic loss due to consumption of feed ingredients highly contaminated with aflatoxins (CAST, 2003). Aflatoxins in poultry reduce feed intake and feed conversion rate which leading to decrease body weight gain, decrease egg production and reproduction performance of both males and females (Hussain et al., 2010). As a common rule, poultry should not get more than 20 μg/kg TA in the feed.

Consumption of food or feed stuffs highly contaminated with aflatoxins can induce severe acute hepatic necrosis which ends by hepatic cirrhosis or hepatocarcinoma. This can result in acute hepatic failure which manifested by disorders in absorption, metabolism of nutrient materials, edema, bleeding and mental abnormalities and coma. Chronic exposure to highly contaminated ingredients with aflatoxins can increase the possibility for hepatocicity and cancer in the gallbladder and by that can result in induction of mutation as the metabolite of aflatoxins intercalate into DNA and alkylate the bases via epoxide moiety (Nogueira et al., 2015).

Aflatoxins are natural toxins that contaminate various types of food and feed stuffs leading to health risk in both human and animals. The aim of this study was to determine the prevalence of aflatoxins contamination and then levels in animal (cattle and poultry) feed samples at Assiut Governorate.

MATERIALS AND METHODS

[1] Samples and sampling: One hundred and forty one feed ingredient (n=58, 26 samples in summer and 32 samples in winter) and complete feeds (n=83, 41 samples in summer and 42 samples in winter) used for cattle and poultry nutrition were collected from Assuit Governorate during winter and summer 2017. The feed ingredient samples were cotton seed cake (n=7, all in winter), maize (n=13, 10 samples in summer and 3 samples in winter), sorghum (n=4, one sample in summer and 3 samples in winter), soybean cake (n=8, 3 samples in summer and 5 samples in winter) and sunflower cake (n=26, 12 samples in summer and 14 samples in winter).

[2] Methods for estimation of aflatoxins levels in examined samples:

Aflatoxins were detected in qualitativly by TLC according to Park et al. (1994) and quantitatively for positive samples by UPLC according to Benvenuti and Burgess (2010).

[3] Chemicals: Aflatoxin standards (B₁, B₂ and total) used in this study were of analytical grade and obtained from Merck Company (Germany).

[4] Apparatus:

- Blender (15000 rpm) with a 1L glass jar and cover.

- Rotary evaporator system Cole-Parmer; Diagonal, 115 VAC

- Shaker, Model EL680, Eberbach Co.

- UV light Chamber with UV lamb at 365 excitation wave length.

- AVICAM Afla Test® Affinity column was used for Cleanup of samples.

- An ACQUITY® UPLC H-class system from Waters Company, USA.

RESULTS

The results of the analyzed samples in this survey were summarized in tables 1-6.

In table 1, out of 7 cotton seed cake samples, one contains TAF (14.285%) and had 300 ppb. Among 13 maize samples, only one had TAF (7.692%) and contains 14 ppb. From 4 sorghum grain samples, 2 were contaminated with AFB1 (50%) and had 12.5±5.303 ppb. Among 8 soybean cake, one (12.5%) contains AFB1 in summer (10 ppb) and 2 in winter (25%) with mean of 4±0.707ppb (3-5). One soybean sample was contaminated with AFB2 (12.5%) and had 3 ppb. From sunflower feed samples (n=26), 11 were contaminated with AFB1, 6 in summer (23.076%) with mean of 89.666±22.188ppb (80-150), and 5 in winter (19.230%) with mean level 78.6±17.226ppb (40-150ppb). 4 samples were contaminated with AFB2, 2 in summer (7.692%) with 54±32.526ppb (8-10) and 2 samples in winter (7.692%) with 77.5±1.767ppb (75-80). 6 samples had TAF, 2 in summer (7.692%) had140.5±77.428 ppb (31-250) and 4 in winter (15.384%) with 65±5.303 ppb.

Table 1: Estimation of aflatoxins levels (ppb) in feed ingredients used for cattle and poultry nutrition.

In table 2, among 30 analyzed cattle feeds, 9 were contaminated with AFB1, 4 with AFB2 and 3 with TAF. In samples contaminated with AFB1, 5 samples were in summer (16.666%) with 15-53 ppb (33.6±6.896) and 4 in winter (13.333%) with 10-75ppb (38.75±13.074). For samples contaminated with AFB2, 2 in summer (6.666%) with 50-53 ppb (51.5±1.060) and 2 in winter (6.666%) with 60-75 ppb (67.5±1.303). 3 samples (11.538%) contaminated with TAF in winter had 70-300 ppb (185±54.211).

In table 2, out of 53 poultry feed samples, 7 had AFB1, 5 in summer (9.433%) with 7.5-30 ppb (13.5±3.714), 2 in winter (3.773%) with 60-70 ppb (65±3.535). 3 samples were contaminated with AFB2, 2 in summer (3.773%) with 7.5-10 ppb (8.75±0.883), 1 in winter (1.886%) had 60 ppb. 3 samples contaminated with TAF. 2 in summer (3.773%) with 16-28 ppb (22±4.424) and 1 in winter had 5 ppb. Among these 53 poultry feeds, 23 samples were for layers, 5 contaminated with AFB1, 3 in summer (13.043%) with 10-30 ppb (16.666±5.443) and 1 in winter (4.347%) with 70 ppb. One sample contaminated with AFB2 in summer (4.347%) which contains 10 ppb. 2 samples contaminated with TAF, 1 in summer (4.347%) which had 16 ppb and 1 in winter (4.347%) which had 15 ppb. For broiler feeds, 3 samples contaminated with AFB1, 2 in summer (7.692%) with 7.5-10 ppb (8.75±0.883) and 1 in winter (3.846%) had 60 ppb. 2 samples contaminated with AFB2, 1 in summer (3.846%) had 7.5 ppb and 1 in winter (3.846%) had 60 ppb. One sample contains 28 ppb TAF in summer.

Table 2: Estimation of aflatoxins levels (ppb) in complete feeds used for cattle and poultry nutrition.

In table 3, for feed ingredient samples, AFB1 and AFB2 were high in summer than in winter. AFB1 in summer samples was 8-150 ppb (78.285±21.742) while in winter was 3-150 ppb (47.333±15.155). AFB2 in summer samples was 8-100 ppb (54±32.526) but in winter were 3-300 ppb (52.666±20.310). For samples contaminated with TAF, winter samples contained high TAF levels ranged from 50-300 ppb (112±42.251) while in summer contained 98.333±62.047 ppb (14-250).

Table 3: Estimation of aflatoxins levels (ppb) in feed ingredients used for cattle and poultry nutrition.

In table 4, for complete feed samples, levels of B1, B2 and TAF was high in winter. B1 in winter samples was 42±11.798 ppb while in summer was 16.562±3.027 ppb. B2 in winter was 65±4.082 ppb while in summer was 36.833±11.996 ppb. TAF in winter samples was 140±56.319 ppb while was 22±4.242 ppb in summer.

Table 4: Estimation of aflatoxins levels (ppb) in complete feeds used for cattle and poultry nutrition.

Table 5: Incidence of the total analyzed samples from feed ingredients and complete feeds used for cattle and poultry nutrition.

Table 6: Estimation of aflatoxins levels (ppb) in all analyzed samples used for cattle and poultry nutrition.