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Background Meat-products are considered an enriched media for mycotoxins. This study aimed to investigate the prevalence of toxigenic Aspergillus species in processed meat samples, HPLC-quantitative measurement of aflatoxin B1 and ochratoxin A residues, and molecular sequencing of aflR1 and pks genes. One hundred and twenty processed beef meat specimens (basterma, sausage, and minced meat; n = 40 for each) were collected from Ismailia Province, Egypt. Samples were prepared for total mold count, isolation, and identification of Aspergillus species. All samples were analyzed for the production of both Aflatoxin B1 and Ochratoxin A mycotoxins by HPLC. Molecular identification of Aspergillus flavus and Aspergillus ochraceus was performed using PCR amplification of the internal transcribed spacer (ITS) region; furthermore, the aflR1 and pks genes were sequenced. Results The total mold count obtained from sausage samples was the highest one, followed by minced meat samples. The prevalence of A. flavus was (15%), (7.5%), and (10%), while the prevalence of A. ochraceus was (2.5%), (10%), and (0%) in the examined basterma, sausage, and minced meat samples, respectively. Using PCR, the ITS region was successfully amplified in all the tested A. flavus and A. ochraceus strains. Aflatoxin B1 was detected in six basterma samples (15%). Moreover, the ochratoxin A was detected only in four sausage samples (10%). The aflR1 and pks genes were amplified and sequenced successfully and deposited in the GenBank with accession numbers MF694264 and MF694264, respectively. Conclusions To the best of our knowledge, this is the first report concerning the HPLC-Molecular-based approaches for the detection of aflatoxin B1 and ochratoxin A in processed beef meat in Egypt. The production of aflatoxin B1 and ochratoxin A in processed meat constitutes a public health threat. Aflatoxin B1 is commonly associated with basterma samples. Moreover, ochratoxin A was detected frequently in sausage samples. The routine inspection of mycotoxins in processed meat products is essential to protect human consumers.

Fungal and mycotoxin contamination of milk products constitute a potential hazard to human health and food safety. Isolation and identifications of mold and yeast out of 140 milk products samples collected from dairy shops in Qena, Egypt were done through conventional microbiological methods. Aflatoxin-M1, aflatoxin-B1 and ochratoxin-A were characterized by thin-layer chromatography; aflR regulatory gene identified by using PCR. Marine algal extracts of Halimeda opuntia, Padina pavonica and Turbinaria decurrens species were studied for their antimicrobial activity. Overall of 80 and 64% dairy products samples were positive for mold and yeast contamination. A total of 38 mold and 15 yeast species were isolated. Aspergillus and Candida spp. were the most abundant isolated species. Furthermore 25, 40 and 27% of cheese and 71, 78 and 73.3 of dairy desserts samples were contaminated with AFM1, AFB1 and OTA, respectively; with average estimated dietary intake level much more than the acceptable daily intake for infant and adult. PCR identified aflR gene among four selected aflatoxigenic A. flavus. The major constituents of H. opuntia extract were 2,4-Decadienal, (E,E)-(21.56%) and 9,12-Octadecadienoic acid (Z,Z)-(36.16%). Ethyl acetate extract of Halimeda opuntia (3mg/ml) exhibited the strongest fungicidal activity with inhibition zones of 16.5 and 22.3 mm against A. flavus and A. niger. It exhibited potent candidacidal activity against C. tropicalis; 11 log10 orders of killing at 750 µg/ml. The discovered antimicrobial activity of H. opuntia is a promising candidate for designing novel antifungal agents which can be used in food preservatives and medicine industry.

This study aimed to evaluate the incidence of fungi in soft skimmed milk cheese and manufacturing of soft skimmed milk cheese from pasteurized milk with trials to control fungal growth by application of some substances (chitosan, natamycin and thyme oil) which known to have antifungal activity . Seventy-five soft skimmed milk cheese samples were collected randomly from supermarkets, groceries, small dairies and street-vendors at Gharbia Governorate for mycological examination. In addition, soft skimmed milk cheese was manufactured from pasteurized milk in lab with addition of natamycin (0.02%, 0.015%), chitosan (1.5%, 1%) and thyme oil (1.5%, 1%) and check the quality of cheeses during storage period. The obtained results revealed that 36% of examined samples were contaminated with mould with a mean count of 4.4×10 3 ±6.8×10 2 cfu/g. The profile of the genera and species of mould isolated from soft skimmed milk cheesesamples were A. flavus 8 (32%), Penicillium spp 14 (56%) P. decumbens 7 (28%), P.citrinum 2 (8%), P. coryphilum 2(8%) and P. concentricum 3(12%) and Byssochlamys nivea 3 (12%). While 68%of examined soft skimmed milk cheese samples were contaminated with yeast with a mean count of 1.8×10 4 ± 2.6×10 3 cfu/g.The frequencydistribution of yeast isolated from the examined soft skimmed milk cheese samples wereCandida22.2% ( C. famta 100% ) Rhodotorulla 19.5% (R. mucilaginosa 42.8% and R. glutinis 57.2%), Saccharomyces13.8%,Debromyceshansenii 19.5%,Yarrowia lipolytica 2.8%and Torulopsis 22.2%. The results of this study showed that manufactured cheese containing natamycin and chitosan had better effect on organoleptic properties and total mould count than cheese containing thyme oil. .

Aflatoxins (AFs), widely distributed food-borne mycotoxins, affect quality and safety of food and cause economic losses in livestock. In this study, the protective effect of Bee Pollen (BP) against some immunotoxic hazards elucidated from eating of AFs-containing diet was investigated in Wistar rats. Rats were randomly classified intofour groups and treated for 30 days, Group 1; control negative, Group 2; Total AFs (3 mg kg(-1) basal diet), Group 3; BP (20 g kg(-1) basal diet) and Group 4; AFs+BP in basal diet. The immunoprotective effect of BP was revealed in terms of increasing (relative to levels seen in Group 2 rats that consumed the AFs diet) serum total protein and globulin levels, restored normal neutrophil (PMN)/lymphocyte ratio, increased PMN phagocytic activity and increased lymphocyte proliferative capacity. Also, the use of the BP reduced spleen H2O2 levels and increased GSH content while maintaining normal levels of NO formation. Histopathologic analysis showed thatthe AFs caused lymphocytic depletion in the spleen; however, BP induced lymphocytic hyperplasia and reduced the levels of AFs-inducible cellular exhaustion or depletion. These results provide evidence of a protective effect of BP against some immunotoxic actions induced in situ by consumption of AFs.