Molecular screening and characterization of Shiga toxin-producing Escherichia coli in retail foods

Shiga toxin-producing Escherichia coli (STEC) strains are one of the most important recently emerged groups of food borne pathogens. This study investigated the prevalence of molecular markers for STEC and characteristics of E. coli O157 isolates from foods sold at retail markets in Wuhan, China. A total of 489 samples (350 meat products and 139 raw vegetables) were purchased from 22 large scale markets between July of 2011 and September of 2013. The meat samples consisted of frozen chicken products, raw pork, raw beef, frozen fish products and processed duck products. The raw vegetable samples consisted of lettuce, bok choy, radish, spinach, cucumber, and tomato. Shiga toxin genes (stx1 and stx2) and an O-group marker of the seven main pathogenic STEC serogroups (O157, O26, O45, O103, O111, O121, and O145) were detected in the samples by using PCR. 100% agreement was obtained between the results of the PCR targeting for wzy_O157 and the PCR targeting for rfbE_O157 gene. The result demonstrated that PCR assay targeting for wzy_O157 gene can be employed as an effective screening method for E. coli O157 in food sample. In the study, E. coli O157 and non-O157 STEC were detected in 55 (11.2%) and 75 (15.3%) samples by PCR screening, respectively. There was significant difference in the occurrence of STEC contamination between supermarkets (19/127, 15.0%) and open markets (111/362, 30.7%) (P < 0.05). Out of 489 samples, 5 samples carried O45, 1 sample carried O145 and 1 sample carried O111. Markers for O103, O26 and O121 were not detected. This result differed from other reports. Immunomagnetic separation based cultivation technique was used to isolate E. coli O157 from 27 food samples collected in 2013. Finally 7 E. coli O157 isolates were obtained. Among the 7 isolates, the prevalent stx genotype was stx_1a and stx_2a. Four E. coli O157 strains exhibited toxic effects on Vero cells, while 3 isolates had no detectable cytotoxicity effects even though they contained stx genes. All E. coli O157 isolates were sensitive to the 12 antimicrobials tested except for roxithromycin. There are some inconsistencies between the PCR screening and culture results. Characteristics of STEC isolates should be evaluated and considered for monitoring STEC contamination in foods.     

Effects of mild and pasteurizing heat treatments on survival of generic and verotoxigenic Escherichia coli from beef enrichment cultures

At many North American beef packing plants, hot water washes and pasteurizing treatments are used to clean and decontaminate carcasses. The aim of this study was to determine whether reduction in numbers of total Escherichia coli found on beef could be regarded as indicative of the reduction in numbers of verotoxigenic E. coli (VTEC) resulting from the same heat treatment. Swab samples were collected from hide-on beef carcasses and were enriched for E. coli in E. coli broth supplemented with novobiocin. Suspensions containing cells in the stationary phase were mixed with equal volumes of meat juice medium and the mixtures were not heated or were heated at temperatures between 55 and 70 °C. All preparations were treated with deoxycholate and propidium monoazide (PMA), then DNA was extracted. Real-time PCR was performed using primers targeting the uidA gene for total E. coli, the stx₁ and stx₂ genes for VTEC, and the eae gene for O157. For samples that were not subjected to heat treatment, cycle threshold (Ct) values were from 13.53 to 17.93 for uidA, 17.94 to 26.77 for stx₁, 21.57 to 29.36 for stx₂, and 23.53 to 28.31 for eae. Ct values for all genes were higher for heat treated than for not treated portions of samples. Differences between Ct values for not treated and heat treated portions of each sample were 5.06–7.57 for uidA, 4.35 to 7.03 for stx₁, 4.49 to 7.12 for stx₂ and 4.75 to 6.77 for eae. Differences between the increases of Ct values for pairs of genes in the sample were 0.32–0.83 for uidA and stx₁, 0.19–0.57 for uidA and stx₂ and 0.31–0.80 for uidA and eae. The maximum change in the ratio of Ct values for uidA and stx₁, stx₂ or eae as a result of heating corresponded to a change of 0.3 log units, which is less than the 0.5 log units generally considered to be microbiologically significant. Therefore, the findings indicate that reductions as a result of mild or pasteurizing heat treatments of total E. coli populations derived from beef can be regarded as indicative of the reductions in sub-populations of O157-VTEC and total VTEC resulting from the treatments.     

Detection of Escherichia coli O157:H7 in ground beef using immunomagnetic separation and multiplex PCR

In the study 251 fresh ground beef samples sold in Ankara were analyzed to evaluate the prevalence of Escherichia coli O157:H7 by immunomagnetic separation (IMS) based cultivation technique. Virulence factors of the isolates were determined by multiplex PCR. Nineteen (7.6%) of 251 ground beef samples were found as contaminated with E. coli O157. By PCR analyse in two of the isolates fliC_h7 gene was detected and identified as E. coli O157:H7. According to the multiplex PCR one of the isolate has all stx₁, stx₂, eaeA, hly and fliC_h7 genes and the other has stx₁, eaeA, hly and fliC_h7 genes.     

Prevalence and characterization of Escherichia coli O157:H7 from samples along the production line in Chinese beef-processing plants

This work investigated the prevalence of Escherichia coli O157:H7 and the antibiotic susceptibility, genetic diversity and virulence genes of isolated strains from four beef slaughter plants in China. A total of 510 samples (feces, hide, carcasses and raw meat) were tested for E. coli O157:H7 using enrichment, immunomagnetic separation, and plating on selective media. The prevalence of E. coli O157:H7 in the feces, hide, pre-evisceration carcass, post-evisceration carcass, post-washing carcass, chilled carcass, and raw meat samples was 1.43%, 1.43%, 0%, 0%, 0%, 1.25%, and 0%, respectively. Multiplex PCR assays were used for serotype confirmation and virulence gene detection. stx₂, eaeA and EHEC-hlyA genes were present in all six of the strains, and the stx₁ gene was not present. Fluorescence amplified fragment length polymorphism (FAFLP) was used to determine the genetic diversity of E. coli O157:H7 and revealed a high similarity between the strains isolated from feces and those isolated from carcasses. None of the isolated strains were found to be resistant to sixteen commonly used antimicrobial agents. The results of this study indicate that although E. coli O157:H7 contamination in the Chinese beef industry is sporadic and not as common as reported in other counties, all of the isolates contained three major virulence genes, presenting a high risk of disease for humans. The current research provides baseline information on E. coli O157:H7 prevalence and character profiles in Chinese beef-processing plants that can be used for future studies.     

Occurrence,serotypes and virulence genes of non-O157 Shiga toxin-producing Escherichia coli in fresh beef,ground beef,and beef burger

This study determined the prevalence, serotypes and virulence genes distribution of non-O157 Shiga toxin-producing Escherichia coli in meat products collected from butchers shops and supermarkets in Mansoura city, Egypt. We have characterized 18 non-O157 STEC strains among the identified 100 E. coli isolates recovered from the examined 87 meat product samples. The prevalence of non-O157 STEC strains in fresh beef, ground beef and beef burger samples were 11.1% (3/27), 16.7% (5/30), and 33.3% (10/30), respectively. The eighteen non-O157 STEC isolated strains were serotyped into seven (38.9%) O111:H8, six (33.3%) O26:H11, two (11.1%) O111:H–, and one (5.56%) for each of O55:H7, O126:H5 and O128:H2. PCR assays for different virulence genes showed that nine (50%), eleven (61.1%), and nine (50%) strains carry stx1, stx2, and eae genes, respectively. The distribution of shiga toxin genes among the isolated strains indicated that seven (38.9%) strains harbored stx1 only, nine (50%) strains harbored stx2 only, and two (11.1%) strains harbored both stx1 and stx2. The eae gene was present in association with five (27.8%), three (16.7%), and one (5.6%) strains that harbored stx1 only, stx2 only, and both stx1 and stx2, respectively. This study concluded that the examined meat products, particularly beef burger, consumed in Egypt are considerably contaminated with a variety of non-O157 STEC serotypes, and hence consumption of such products may constitute a potential health risk for consumers.     

Prevalence assessment of Coxiella burnetii and verocytotoxin-producing Escherichia coli in bovine raw milk through molecular identification

Raw milk has become increasingly appreciated by consumers and in the EU there is general acceptance of this product, the sale of which is regulated by EU Directives. However, unpasteurised milk can be contaminated by zoonotic agents, thus representing a risk for human health. The aim of our work was the assessment of Coxiella burnetii and verocytotoxin-producing Escherichia coli (VTEC) presence in bovine bulk tank milk of Marche region (Central Italy) destined to direct human consumption. PCR and real-time PCR well-established protocols have been used for pathogen identification, showing high sensitivity and specificity. C. burnetii prevalence was 27%, while VTEC serogroup were found as follows: 3.5% for both O157 and O145; 2.3% for O26; 4.7% for O103. In contrast, the immunomagnetic separation-based protocol (IMS) was able to identify only one sample (1.2%) contaminated by VTEC O157.These results indicate that a combination of real-time PCR and IMS proved to be more appropriate for VTEC serogroup identification, thanks to improved sensitivity. Moreover, the prevalence of these pathogens would suggest that non-O157 VTEC and C. burnetii should be included among microbiological criteria for raw milk, implementing control strategies to limit possible negative impact to consumers' health.     

Development of a real-time PCR procedure for quantification of viable Escherichia coli in populations of E. coli exposed to lactic acid,and the acid tolerance of verotoxigenic E. coli (VTEC) from cattle hides

The aims of this study were to develop a real-time PCR procedure for determining the effects on Escherichia coli of treatment for decontaminating beef carcasses with lactic acid solution, and determining if there were differences in the acid tolerance of E. coli generally and verotoxigenic E. coli (VTEC). Suspensions of E. coli were incubated with 4% lactic acid at pH 3.6. The numbers of surviving E. coli at different incubation times were determined from plate counts and from quantification by real-time PCR of the uidA gene in DNA preparations. The numbers of viable E. coli progressively declined, by about 4 log units during incubation for 6 h. The mean cycle threshold (Ct) values for uidA in DNA from samples collected at different times and treated or not treated with propidium monoazide (PMA) before DNA extraction were similar. Treatment with 1% sodium deoxycholate (SD) before PMA treatment resulted in an increase of >6 Ct when the reduction in viable cell number was around 1 log. When E. coli incubated with 4% lactic acid solutions of pH 2.4, 2.8, 3.2 or 3.6 were resuscitated in half strength brain heart infusion (BHI) for 2 h before treatments with SD and PMA, the slope of the plot relating Ct values to the numbers of viable E. coli was 1.85 Ct log cfu⁻¹ with the correlation coefficient (R²) being 0.80. The findings indicate that while the membranes of E. coli inactivated by 4% lactic acid were largely impermeable to PMA, the membranes of both dead and injured cells were rendered permeable to PMA by treatment with 1% SD. Resuscitation in BHI restored the membrane barrier properties of the injured cells. Treatment with lactic acid resulted in increases in Ct values of 4.1, 3.7, 2.5 and 1.5 for the uidA, stx₁, stx₂ and eae genes, respectively; and the increases in Ct values for the latter two genes were significantly different (p < 0.05) from that for the uidA gene. This indicates that VTEC carrying stx₂ and/or eae were more acid resistant than other E. coli. Thus, caution should be exercised when using generic E. coli as an indicator for VTEC for assessment of the antimicrobial efficacy of organic acid decontaminating treatments at abattoirs.     

Incidence of Listeria monocytogenes in beef,pork, raw-dried and raw-smoked sausages in Bulgaria

For a 5-years period (2002–2007) 786 samples (505 samples of beef, pork, minced beef, minced pork and 281 samples of raw-dry and raw-smoked sausages) were analyzed. From beef and pork 39 strains Listeria monocytogenes (7.7%), three strains Listeria ivanovii (0.6%), 23 strains Listeria innocua (4.6%) and four strains Listeria welshimeri (0.8%) were obtained. L. monocytogenes were isolated in 28 samples (10.0%), L. ivanovii, L. innocua and L. welshimeri – in 0.7%, 4.3% and 0.7%, respectively, from investigated raw-dried and raw-smoked sausages.     

Seasonal study of aflatoxin M1 contamination in milk of four dairy species in Yazd,Iran

This survey was conducted to determine the occurrence of aflatoxin M₁ (AFM₁) in samples of raw milk obtained from cow, sheep, goat, and camel herds in Yazd province during different seasons. Aflatoxin M₁ was analyzed using the competitive enzyme-linked immunosorbent assay technique for screening and high-performance liquid chromatography with fluorescence detection for confirmatory purposes. The detection rates of AFM₁ in cow, sheep, goat, and camel milk samples were 46.5%, 21.6%, 20.1%, and 4.03%, respectively. Levels of the toxin in 15.4% of cow milk, 11.5% of sheep milk, and 9.15% of goat milk samples exceeded the legal limit (0.050 μg/kg) recommended by the Institute of Standards and Industrial Research of Iran; while none of the camel milk samples exceeded the legal limit. The occurrence and levels of AFM₁ in cow milk samples from industrial dairy farms was significantly lower (P ≤ 0.05) than those from traditional ones. Seasonal variations influenced the occurrence and levels of AFM₁ in cow, sheep, and goat milk; however, no statistically significant seasonal effect was found for camel milk. This study indicates a high occurrence of AFM₁ in cow milk especially those obtained from traditional dairy farms. Therefore, more supervision is required on these farms; and traditional dairy farms should be gradually replaced by industrial ones.     

Prevalence of Yersinia enterocolitica from food and pigs in selected states of Malaysia

The aim of this study was to determine the prevalence of Yersinia enterocolitica and its bioserotypes from food and pigs in Malaysia. Fifty-eight raw porcine (raw pork meat, internal organs and other parts) and 48 non-porcine food (raw beef, poultry products, seafood, vegetables, tofu, and pasteurised milk) from wet markets located in Kuala Lumpur, Selangor, Perak, and Pahang were examined for the presence of Y. enterocolitica. Specimens (nasal, oral and rectal swabs) from 165 pigs (from nine farms) located at central and northern parts of Malaysia were also collected for Y. enterocolitica detection. Presumptive isolates were characterised biochemically and further confirmed by PCR. Out of 58 raw porcine food, Y. enterocolitica was detected in 7 (12.1%) samples in which raw pork meat (whole meat) had the highest prevalence 5/21 (23.8%), followed by raw pork liver 1/5 (20.0%) and raw pork intestine 1/8 (12.5%). No Y. enterocolitica was isolated from the 48 non-porcine foods. Overall, two pathogenic (bioserotypes 3 variant/O:3 and 1B/O:8) and one non-pathogenic (bioserotype 1A/O:5) Y. enterocolitica strains were isolated from food. Out of 165 pigs examined, 3 (1.8%) pigs were carriers for Y. enterocolitica. All 3 pigs were asymptomatic grower pigs from Penang, carried Y. enterocolitica bioserotype 3 variant/O:3. Post-enrichment PCR approach gave a higher prevalence, 60.3%, 41.7% and 27.9% for porcine food, non-porcine food and pigs, respectively. Both pathogenic and non-pathogenic Y. enterocolitica were present in our domestic pigs and food. Improper food handling and processing may cause cross contamination of this pathogen to humans, affirms a potential risk for public health.     

Development of a controlling method for Escherichia coli O157:H7 and Salmonella spp. in fresh market beef by using polylysine and modified atmosphere packaging

Escherichia coli O157:H7 and Salmonella spp. often contaminate fresh beef. In Japan, an E coli outbreak caused by raw beef made 181 people ill and 5 individuals dead in 2011. Responding to this outbreak, an effective sterilization method for fresh beef is expected to be developed. In this study, ε-polylysine combined with CO₂-packaging method was examined for controlling these pathogens in fresh beef. At an incubation temperature of 4 °C, approximately 4.3 log and 2.4 log reduction in bacterial numbers were observed after 7-day incubation for E. coli O157:H7 and Salmonella, respectively, in ε-polylysine-added beef. When effectiveness of CO₂-packaging combined with ε-polylysine was investigated, CO₂ did not have additional inhibiting effect on bacterial growth compared to only-ε-polylysine-treated samples when incubated at 4 °C. However, effectiveness of CO₂ was observed when incubated at 10 °C where approximately 2.9 log and 4.4 log reduction in E. coli cell numbers were observed in only-ε-polylysine-treated samples and polylysine- and CO₂-treated group, respectively, and approximately 1.7 log and 3.5 log reduction in Salmonella cell numbers were observed in only-ε-polylysine-treated samples and polylysine and CO₂-treated group, respectively. This study confirmed that ε-polylysine or ε-polylysine combined with CO₂ packaging are effective in preventing foodborne diseases caused by raw beef.     

Prevalence and antimicrobial resistance of Listeria species isolated from milk and dairy products in Iran

The objective of this study was to determine the prevalence of Listeria spp. in milk and dairy products in Isfahan province, Iran. From March 2007 to September 2009, a total of 594 samples of various milk and dairy products were obtained from randomly selected retail stores. Using conventional bacteriologic method, 55 samples (9.3%) were positive for Listeria spp. The highest prevalence of Listeria was found in raw sheep milk samples (22.6%), followed by cheese samples (18.9%). The most species recovered was Listeria innocua (58.2%); the remaining isolates were Listeria monocytogenes (32.7%) and Listeria seeligari (9.1%). Overall, 54 Listeria isolates (98.2%) were resistant to one or more antimicrobial agents. Resistance to nalidixic acid was the most common finding (96.4%). All Listeria isolates were susceptible to vancomycin. The results of this study indicate the potential risk of infection with Listeria in people consuming raw and unpasteurized milk and dairy products.     

Occurrence of aflatoxin M1 in raw milk in South Korea using an immunoaffinity column and liquid chromatography

The level of aflatoxin M₁ (AFM₁) contamination in raw milk produced in South Korea was investigated using immunoaffinity column chromatography and high performance liquid chromatography with fluorescence detector. A total of 100 raw milk samples were collected from 100 cattle ranches located in three different provinces of South Korea. Forty eight out of 100 raw milk samples contained AFM₁ at low level (0.002–0.08 μg/L) with mean value of 0.026 μg/L. Among the AFM₁ contaminated samples, 29 raw milk samples contained only traceable amount of AFM₁ below the limit of LOQ, 0.02 μg/L. None of samples exceeded the maximum level (0.5 μg/L) of Korean regulation for AFM₁ in milk. The limit of detection was 0.002 μg/L. The result of recovery test with 0.5 μg/L AFM₁ in raw milk sample was 96.3% (SD 3.6, n = 5). This is the first pioneering study to investigate the level of AFM₁ in raw milk used in dairy industries in South Korea.     

Incidence and genetic variability of Listeria species from three milk processing plants

The presence of Listeria in three milk processing environments as a potential source of milk contamination was assessed. Swab samples (n = 210) taken from milk processing plants were examined. Sample sites included the milk processing equipment, besides areas handling raw and pasteurized milk. The USDA Listeria-selective enrichment procedure was used to process the samples. Forty one (19.52%) Listeria isolates were recovered. The isolates were further subjected to biochemical and genotypic characterization. Out of 41 isolates, 16 (7.62%) were confirmed as Listeria monocytogenes, 2 (0.95%) as L. ivanovii, 19 (9.05%) as L. innocua. 1 (0.48%) as L. seeligeri and 3 (1.43%) as L. grayi. All the L. monocytogenes isolates were positive for the hlyA gene. PCR based serotyping revealed all L. monocytogenes to be of 1/2a, 1/2c, 3a and 3c serovar group. AscI and ApaI restriction analysis yielded four PFGE clusters for 16 L. monocytogenes isolates obtained from raw milk collector, milk silos, buttermilk mixer, cheese and other milk product processor. No predominant PFGE cluster was observed among these L. monocytogenes isolates. The main sources of L. monocytogenes were found to be raw milk collector and milk silos. In the present study L. monocytogenes was isolated from milk and milk products processing plants which could cross-contaminate the processed products and may possess a potential threat to public health.     

Aflatoxin M1 in milk and traditional dairy products from west part of Iran: Occurrence and seasonal variation with an emphasis on risk assessment of human exposure

In the present study, a total of 358 samples consisting of raw milk of cow (n = 64), goat (n = 56) and sheep (n = 52); traditional cheese (n = 40), yoghurt (n = 42), Kashk (n = 40), Doogh (n = 44) and Tarkhineh (n = 20) were analyzed for aflatoxin M₁ (AFM₁) by using an enzyme linked immunosorbent assay (ELISA). Frequency of AFM₁ and its concentration ranges in the ELISA positive samples were determined by high performance liquid chromatography with fluorescence detection (HPLC-FD). AFM₁ contamination was 84.3%, 44.6% and 65.3% for cow, goat and sheep raw milks, respectively. Moreover, AFM₁ was in 65.5%, 23.8%, 14%, 13.6% and 35.0% of cheese, yoghurt, Kashk, Doogh and Tarkhineh samples, respectively. Percentages of cow milk, goat milk, sheep milk and cheese samples exceeding the EU limit were 35.9%, 11.1%, 26.9% and 10%, respectively. HPLC analyses confirmed the ELISA results although the percentages of AFM₁ contamination in raw milk and dairy products were lower than that of ELISA. There were significant differences (P < 0.05) between the mean AFM₁ contents of raw milk, cheese and yoghurt samples during winter and summer seasons. Our study demonstrated that there is a potential risk for liver cancer due to the consumption of milk and dairy products in Iranian consumers.     

Prevalence and antimicrobial resistance of Staphylococcus aureus isolated from raw milk and dairy products

The objectives of this study were to determine the prevalence and antimicrobial resistance of Staphylococcus aureus isolated from raw milk (cow and sheep) and dairy products (traditional cheese and kashk) in Mazandaran Province, Iran. A total of 2650 samples, including 1930 raw milk and 720 dairy products were purchased from retail stores. Out of 2650 samples, S. aureus was detected in 328 samples (12.4%) in which 53 (16.2%) were positive for methicillin-resistant S. aureus. The S. aureus isolates showed resistance to tetracycline (56.1%), followed by penicillin G (47.3%), oxacillin (16.2%), lincomycin (11.9%), clindamycin (11.3%), erythromycin (7.9%), streptomycin (5.8%), cefoxitin (5.5%), kanamycin (4%), chloramphenicol (3.7%), and gentamicin (2.1%). A high frequency of blaZ (46%) and tetM (34.8%) resistance genes was found in S. aureus isolates. The findings of this study revealed consumption of raw milk and dairy products as a potential risk of foodborne infection in this region.     

Seasonal variation of aflatoxin M1 in raw milk from the Yangtze River Delta region of China

The objective of this study was to evaluate the occurrence of aflatoxin M₁ (AFM₁) in raw milk samples from 18 dairy farms in the Yangtze River Delta region during four different seasons. A total of 72 tank milk samples was collected with 18 samples for each season. Milk AFM₁ was detected using LC-MS/MS. The AFM₁ was detected in 43 milk samples (59.7%) ranging in concentration from 10 to 420 ng/L. The concentration of AFM₁ in raw milk was significantly higher during the winter (123 ng/L) than during other seasons (P < 0.05). There was no significant difference between the spring (29.1 ng/L), summer (31.9 ng/L), and autumn (31.6 ng/L) (P > 0.05) seasons. This indicates that raw milk collected during the winter is at high risk for AFM₁ and that seasonal factors should be considered for the management of aflatoxins in both the feed and milk.     

Incidence of Listeria monocytogenes and Escherichia coli O157:H7 in two Kasar Cheese processing environments

This study was designed to determine the presences of two environmental pathogens in two dairy factories in Sakarya, Turkey. A total of 264 environmental samples, raw milk and cheese samples were taken at four different seasons. According to the results, Listeria monocytogenes or Escherichia coli O157:H7 was isolated from 26 or 2.7% of the samples collected from both factories, respectively. None of the cheese or curd samples were found to be positive for Listeria or E. coli O157:H7. However, 50% of raw milk samples contained Listeria innocua. Listeria was mostly isolated from the swap samples taken from the drains or the floors in processing or packaging areas. However, E. coli was also isolated from the swap samples taken from the workers’ hands and gloves as well as the drains and the floor. Only one raw milk sample contained E. coli O157:H7. A higher prevalence of both pathogens was observed in the summer months than in the other months.     

Variation of aflatoxin M1 contamination in milk and milk products collected during winter and summer seasons

Total 221 samples of milk and milk products were collected during winter (November 2011–February 2012) and 212 samples were collected during summer (May–August 2012) from central areas of Punjab, Pakistan. The samples were analyzed for the presence of aflatoxin M₁ (AFM₁) with a validated HPLC method equipped with florescence detector. The results revealed that from winter season almost 45% samples of milk and milk products were found to be contaminated with AFM₁ i.e. 40% of raw milk, 51% of UHT milk, 37% of yogurt, 60% of butter and 43% of ice cream samples and 27, 24, 25, 34 and 17% of samples were found above the recommended limit for AFM₁, respectively. However, from summer season 32% samples of milk and milk products were found to be contaminated i.e. 36% of raw milk, 31% of UHT milk, 29% of yogurt, 40% of butter and 24% of ice cream and 23, 23, 18, 20 and 5% of samples were found above the permissible limit for AFM₁, respectively. The levels of contamination in winter milk and milk product samples were significantly higher (α ≤ 0.05) than in summer season. The occurrence of AFM₁ in milk and milk products were higher, demanding to implement strict regulations and also urged the need for continuous monitoring of milk and milk products in order to minimize the health hazards.