Identification of Dairy Fungal Contamination and Reduction of Aflatoxin M1 Amount by Three Acid and Bile Resistant Probiotic Bacteria.

Aflatoxins (AFs) released by fungi are observed in the cow’s milk even after pasteurization. Aflatoxin M1 (AFM1) has particularly an incredible clinical significance, as a critical carcinogenic agent for humans. Several strategies have been implemented for lowering the AFM1 amount, such as the employment of probiotics, particularly lactobacilli or lactic acid bacteria (LAB). However, this strategy has not been applied routinely until today. This study aimed to evaluate the effect of three LABs on the reduction of AFM1 in traditional milk and cheese samples. In total, 85 milk (n=45) and cheese (n=40) samples were obtained from the open markets of Shiraz, Iran, from February to June 2018. Additionally, the AFM1 levels were evaluated, compared to those of the National Iranian Standard. The data were then analyzed in SPSS software (version 20) through the Chi-square test. Statistical analysis was performed at a 95% confidence level (p-value of <0.00001). Out of 50 purchased LABs, the efficient antifungal property and resistance to bile salts were observed in five strains. The mean value of these five strains was calculated after adding 5 ppm AFM1, compared to natamycin. The strains with a reduction in AFM1 level were sequenced and registered in the NCBI database.In total, 15 samples with contamination higher than the allowed limit included Penicillium spp, Aspergillus niger, Saccharomyces cerevisia, Saccharomyces paradoxus, and Yarrowia lipolytica.The results also showed reduced AFM1 levels in three LAB-treated strains. Lactobacillus fermentum CECT562 (T), Lactobacillus brevis ATCC14869 (T), and Enterococcus faecium LMG 11423 (T) had this capability to 0.05, 0.03, and 0.03 respectively. The National Iranian Standard should be implemented to have control over traditional dairy products with more care. The three LABs selected in the current study revealed a significant effect on reducing AFM1 levels in traditional milk and cheese.

Aflatoxin M1-Binding Ability of Selected Lactic Acid Bacteria Strains and Saccharomyces boulardii in the Experimentally Contaminated Milk Treated with Some Biophysical Factors.

There is a growing concern regarding the recurrent observation of aflatoxins (AFs) in the milk of lactating animals. Regarding this, the present study was conducted to assess the aflatoxin M1 (AFM1)-binding ability of three species, namely Lactobacillus rhamnosus, L. plantarum, and Saccharomyces boulardii, inAFM1-contaminatedmilk. The mentioned species were administeredatthe concentrations of107 and 109 CFU/mLto skimmed milk contaminated with 0.5 and 0.75 ng/mL AFM1 within the incubation times of 30 and 90 min at 4°C and 37°C. Lactobacillus rhamnosus was found to have the best binding ability at the concentrations of 107 and 109 (CFU/ml), rendering 82% and 90% removal in the milk samples with 0.5 and 0.75 ng/ml AFM1, respectively. Accordingly, this value at 107 and 109 CFU/ml of L. plantarum was obtained 89% and 82% with 0.75 ng/ml of AFM1, respectively. For S. boulardii at 107 and 109 CFU/ml, the rates were respectively estimated at 75% and 90% with 0.75 ng/ml of AFM1. The best AFM1-binding levels for L. rhamnosus, L. plantarum, and S. boulardii were 91.82±10.9%, 89.33±0.58%, and 93.20±10.9, respectively, at the concentrations of 1×109, 1×107, and 1×107 CFU/ml at 37, 4, and 37°C, respectively. In this study, the maximum AFM1 binding (100.0±0.58) occurred while a combination of the aforementioned probiotics was employed at a concentration of 1×107 CFU/ml at 37°C with 0.5 ng/ml AFM1, followed by the combination of L. rhamnosus and L. plantarum (95.86±10.9) at a concentration of 1×109 CFU/ml at the same temperature with 0.75 ng/ml AFM1. It was concluded that the use of S. boulardii in combination with Lactobacillus rhamnosus and L. plantarum, which bind AFM1 in milk, can decrease the risk of AFM1 in dairy products.

VacA and cagA genotypes status and antimicrobial resistance properties of Helicobacter pylori strains isolated from meat products in Isfahan province, Iran.

Although Helicobacter pylori has a significant impact on the occurrence of severe clinical syndromes, its exact ways of transmission and origin have not been identified. According to the results of some previously published articles, foods with animal origins play a substantial role in the transmission of H. pylori to humans. The present investigation was carried out to study the vacuolating cytotoxin A (vacA) and cytotoxin associated gene A (cagA) genotypes status and antibiotic resistance properties of H. pylori strains recovered from minced-meat and hamburger samples. A total of 150 meat product samples were collected from supermarkets. All samples were cultured and the susceptive colonies were then subjected to nested-PCR, PCR-based genotyping and disk diffusion methods. 11 out of 150 samples (7.33%) were positive for H. pylori. All the isolates were further identified using the nested-PCR assay. Prevalence of H. pylori in hamburger and minced-meat samples was 1.42% and 12.5%, respectively. S1a, m1a and cagA were the most commonly detected genotypes. The most commonly detected combined genotypes in the H. pylori strains of minced-meat were s1am1a (10%), s1am1b (10%) and s2m1a (10%). Helicobacter pylori strains of meat products harbored the highest levels of resistance against ampicillin (90.90%), erythromycin (72.72%), amoxicillin (72.72%), trimethoprim (63.63%), tetracycline (63.63%), and clarithromycin (63.63%). Hamburger and minced-meat samples may be the sources of virulent and resistant strains of H. pylori. Meat products are possible sources of resistant and virulent strains of H. pylori similar to those vacA and cagA genotypes. Using healthy raw materials and observation of personal hygiene can reduce the risk of H. pylori in meat products.

Prediction of Listeria spp. growth as affected by various levels of chemicals, pH, temperature and storage time in a model broth.

The effects of concentration of NaCl (0.5 to 12.5%), methyl paraben (0.0 to 0.2%), sodium propionate (0.3%), sodium benzoate (0.1%), potassium sorbate (0.3%), pH (> 5.9) temperature (4 to 30 degrees C), storage time (up to 58 d) and inoculum (> 10(5) to > 10(-2) per ml) on the log10 probability percentage of one cell of Listeria spp. to initiate growth in a broth system were evaluated in a factorial design study. At pH 5.96 and temperature ranging from 4 to 30 degrees C the concentrations of sodium propionate, potassium sorbate, and sodium benzoate examined allowed growth of L. monocytogenes with lag phases at 4 degrees C of 18, 27 and 21 days, respectively. For 0.1 and 0.2% methyl paraben growth of all Listeria spp. was initiated at 8 degrees C and 30 degrees C, respectively. At pH 6, concentration of 12% NaCl supported the growth of L. monocytogenes at 8 to 30 degrees C, whereas 12.5% inhibited all Listeria species. Four regression equations were derived relating probability of growth initiation to temperature, concentrations of NaCl and preservatives storage time, and Listeria species specific effects. From these equations, the number of cells needed for growth initiation can be calculated. The impact of this type of quantitative study and its possible application on the development of microbial standards for foods is discussed.

Growth and toxigenesis of C. botulinum type E in fishes packaged under modified atmospheres.

Modified atmosphere packaging of fresh fish is used to market high quality products in some European countries. The potential risk of C. botulinum growth in these extended shelf-life foods is still a concern; especially since toxigenesis may precede organoleptic spoilage. This paper will present toxigenic data from rockfish, salmon and sole muscle tissues which were inoculated with a pool of non-proteolytic C. botulinum type E at seven levels (10(-2)-10(4) spores/sample), and stored under vacuum and 100% CO2, at incubation temperatures between 30 and 4 degrees C, for up to 60 days. Factorial experimental design allowed predictive formulae to be developed able to describe the lag time prior to C. botulinum toxigenesis and the probability of one spore to initiate toxigenesis based upon the storage conditions. Accurate characterization of the microbial ecology of C. botulinum in modified atmosphere-packaged fish, will support safe exploitation of these packaging systems in the market place, and identify critical control points for potential product or process abuses.