Occurrence and exposure assessment of Aflatoxin M1 in milk and milk products in India.

Milk containing Aflatoxin M1 (AFM1) poses a serious health risk to consumers. Present study was undertaken to determine levels of AFM1 in 146 milk and value added dairy products sold in retail markets of Chhattisgarh, India using HPLC coupled with fluorescence detector. A total of 52 samples (35.6%) were found to contain AFM1 with overall concentrations ranging from nd - 2.608 µg/L. The contamination levels were higher in non-fermented milk products than fermented milk products samples, although this difference was statistically non-significant (p > 0.05). AFM1 concentrations above maximum permissible limits established by the European Commission were found in 94.2% of positive samples. Health risk assessments ascertained that the estimated daily intakes for AFM1 is higher than the established tolerable daily intakes for both adults and children (Hazard Index > 1), there by implying a potentially high risk to consumer's health. Current investigation provides valuable information regarding contamination of raw as well as value added milk products sold in Indian markets. Therefore, to protect consumer's health and promote dairy trade; there is an urgent need to increase farmer's knowledge on good storage practices of feed and fodder. Further, stringent enforcement of food safety regulations is imperative to safeguard and promote human health.

Detection of aflatoxin M1 in bovine milk from different agro-climatic zones of Chhattisgarh, India, using HPLC-FLD and assessment of human health risks.

Concerns regarding food safety and 'One Health' are increasing globally. Aflatoxin M1 (AFM1), a human carcinogenic toxin, is excreted by lactating animals in their milk after consumption of feed contaminated with aflatoxin B1. The present cross-sectional study aimed to determine the occurrence of AFM1 in cattle and buffalo milk produced in rural and peri-urban areas under different agro-climatic conditions of Chhattisgarh, India, and assesses human health risks. Analyses of 545 milk samples by validated high-performance liquid chromatography revealed high level of AFM1 contamination in 224 (41.1%) samples with mean concentration of 0.137 ± 0.029 µg/L. Statistically significant differences (p< 0.05) were observed in the levels and frequency of AFM1 occurrence among different agro-climatic zones. AFM1 was more frequently detected in milk samples from Northern hills (64%) followed by Bastar plateau (40.7%) and Chhattisgarh plain (27.3%), with mean concentration levels of 0.396 ± 0.099 µg/L, 0.081 ± 0.025 µg/L and 0.013 ± 0.002 µg/L, respectively. Species wise no significant difference was observed in the detection frequency and concentration of AFM1 in milk from cattle and buffalo. AFM1 contamination above maximum permissible limits established by European commission and Food Safety and Standard Authority of India was detected in 21.3% and 4.4% of samples, respectively. The estimated daily intakes for AFM1 were found to be higher than tolerable daily intakes for both adults and children, especially of Northern hills implying a potentially high risk to consumer's health. This study provides valuable information on the contamination status of milk in one of the fastest developing state of India. It also highlights the importance and need for continuous farmers' awareness on good animal husbandry practices, routine surveillance of mycotoxins in animal feeds and food commodities to safeguard human health.

Assessment of aflatoxin contamination in dairy animal concentrate feed from Punjab, India.

Aflatoxins are one of the major environmental contaminants in animal feed and pose a potential threat to human health due to their secretion in the milk of lactating animals. The present study was conducted with the objectives to determine the occurrence of aflatoxins (B1, B2, G1, and G2) in dairy animal concentrate feed and to evaluate the effect of season, spatial variation, and dairy farm size on the levels of aflatoxins contamination. A total of 189 dairy animal concentrate feed samples were tested for aflatoxins with enzyme-linked immunosorbent assay (ELISA) as screening and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) as confirmatory techniques. Of the total, 59% feed samples were found positive for aflatoxins, while 44% samples were detected with total aflatoxins levels higher than the tolerance limit established by the Food and Drug Administration (FDA) and 58% samples were found with aflatoxins B1 (AFB1) levels above the European Commission (EC) legal limit. AFB1 levels in dairy animal concentrate feed were found significantly higher during rainy (41.6 µg kg-1) and winter (35.9 µg kg-1) seasons as compared to the summer season (25.5 µg kg-1). The theoretical extrapolation of the AFB1 carry-over from animal feed to milk (aflatoxins M1) in different seasons may lead to 50-100% contamination of milk at levels above the EC tolerance limit. The incidence and levels of aflatoxins especially AFB1 in animal feed, not only pose a direct effect on animals but may also pose a concern for food safety in relation to the occurrence of aflatoxins M1 in milk. Therefore, continuous surveillance of aflatoxins in dairy animal feeds is required to reduce animal and consequently human exposure.

Potential risk factors associated with the occurrence of aflatoxin M1 in raw milk produced under different farm conditions.

The present study was carried out with the objectives to determine the occurrence of aflatoxin M1 (AFM1) in raw milk and identification of associated risk factors from three different types of animal farming systems. A total of 189 pooled raw milk samples were tested for AFM1 with enzyme linked immunosorbant assay (ELISA) as screening and high performance liquid chromatography with fluorescence detection (HPLC-FLD) as confirmatory techniques. AFM1 was found positive in 58% pooled milk samples with the mean level of 0.917 µg/L, while 50.8% and 36.5% samples were found above the legal limits set by European Commission (EC) and Food safety and standard authority of India (FSSAI), respectively for AFM1 in milk. Among all investigated risk factors, feeding of readymade concentrate feed and leftover household cereals, longer feed storage duration and feed storage quality were found significantly associated with presence of AFM1 in farm milk. High prevalence as well as levels of AFM1 in farm milk poses a health risk to the consumers especially children. Therefore, efforts are urgently required to create awareness among dairy farmers about aflatoxins contamination of feed and milk, to improve farm management practices and to control AFM1 level in milk below the legal limits.

Characterization of Salmonella Gallinarum from an outbreak in Raigarh, Chhattisgarh.

AIM: The present investigation was conducted to isolate and characterize Salmonella Gallinarum from an outbreak of fowl typhoid in layer birds. MATERIALS AND METHODS: Clinically ill and dead layer birds from an outbreak were investigated. History, clinical signs, and postmortem lesions were suggestive of fowl typhoid. Postmortem samples including heart blood, intestinal contents, pieces of ovary, and liver were collected and processed immediately for bacterial culture, serotyping and antibiotic sensitivity tests. Isolates were further screened for the presence of extended spectrum beta lactamase (ESBL) (blaTEM) gene by polymerase chain reaction. RESULTS: On the basis of cultural, staining and biochemical characteristics; three bacterial isolates were confirmed as S. Gallinarum. On serotyping, somatic antigen O: 9 and 12 with nonflagellated antigen were detected in all three isolates. Isolates were intermediate sensitive to amoxycillin, amoxyclav, gentamicin and ciprofloxacin and resistant to most of the antibiotics including chloramphenicol, ampicillin, ceftazidime, cefexime, cefepime, azithromycin, nalidixin, tetracycline, oxytetracycline, and streptomycin. Two isolates were found to harbor ESBL (blaTEM) gene. CONCLUSION: Beta lactamase producer S. Gallinarum was confirmed as cause of increased mortality in layer birds during present investigation. Existence of multi drug resistant Salmonella poses serious threat to poultry industry in Chhattisgarh.

Occurrence and characteristics of extended-spectrum ß-lactamases producing Escherichia coli in foods of animal origin and human clinical samples in Chhattisgarh, India.

AIM: To assess the prevalence of antimicrobial resistance producing extended-spectrum ß-lactamases (ESBL) (blaTEM, blaSHV, and blaCTX-M) genes in Escherichia coli isolated from chicken meat, chevon meat, raw milk, and human urine and stool samples collected from tribal districts of Chhattisgarh, viz., Jagdalpur, Dantewada, Kondagaon, and Kanker. MATERIALS AND METHODS: A total of 330 samples, comprising 98 chicken meat, 82 chevon meat, 90 raw milk, and 60 human urine and stool samples, were processed for isolation of E. coli. Isolates were confirmed biochemically and further tested against commonly used antibiotics to know their resistant pattern. The resistant isolates were tested for ESBL production by phenotypic method followed by characterization with molecular method using multiplex-polymerase chain reaction technique. RESULTS: Overall 57.87% (191/330) samples were found positive for E. coli, which include 66.32% (65/98) chicken meat, 46.34% (38/82) chevon meat, 81.11% (73/90) raw milk, and 25% (15/60) human urine and stool samples. Isolates showed the highest resistance against cefotaxime (41.36%) followed by oxytetracycline (34.03%), ampicillin (29.31%), cephalexin (24.60%), cefixime (16.75%), and ceftazidime (13.08%). Phenotypic method detected 10.99% (21/191) isolates as presumptive ESBL producers, however, molecular method detected 3.66% (7/191), 2.09% (4/191), and 0.00% (0/191) prevalence of blaTEM, blaCTX-M, and blaSHV, respectively. CONCLUSION: The present study indicates a high prevalence of E. coli in raw chicken meat, chevon meat, and milk due to poor hygienic practices. The antibiotic susceptibility test detected the presence of the resistance pattern against ESBL in E. coli isolated from raw chicken meat, chevon meat, milk, and also in human clinical samples is of great concern. The appearance of E. coli in the human food chain is alarming and requires adaptation of hygienic practices and stipulate use of antibiotics.