Bovine subclinical mastitis was one of the most important health problems facing dairy industry, its impact exceeded the economic aspects and extended to potential negative effects on human health. The current study aimed to determine the prevalence of E. coli as an important mastitic pathogen and identify some of its most important virulence gene as well as their antimicrobial resistance profile. In the present study E. coli was isolated and biochemically identified whereas out of 100 subclinically mastitic milk samples was nine samples were positive for E. coli with 9% prevalence rate. Serotyping of these isolates declared that 3 isolates were serotype O26:H11, 2 isolates in serotype O91:H21 and 1 isolate in each of serotypes O55:H7, O128:H2, O146:H21 and O124. Antimicrobial resistance profile of the obtained isolates showing that all the isolates were 100% resistant to both erythromycin and streptomycin, while 88.9% (8/9) were sensitive to gentamicin. The presence of 3 important virulence factors including shiga toxin1(stx1), shiga toxin 2 (stx2) and intimin (eae) genes, among the obtained isolates was reported using PCR. Molecular investigation revealing that 2 isolates contain all studied virulence genes (stx1, stx2 and eae), 3 isolates contain (stx1 and stx2), while stx1 was detected solely in 2 isolates, also 1 isolate contain only stx2 and lastly 1 isolate was negative for any of the studied virulence factors. In a conclusion, there was a 9% prevalence rate of E. coli in subclinically mastitic milk samples in the current study, indicating its importance as a mastitic pathogen. The shiga toxin genes (stx1 & stx2) are widely distributed among E. coli isolates, while the intimin (eae) gene is less prevalent in comparison to shiga toxin genes. Also the recorded high multidrug resistance rate among the isolates posing threat to human health though entrance of these strains into the human being food chain whereas the isolated E. coli strains had the highest resistance to erythromycin and Streptomycin (100%), followed by Clindamycin (77.8%), Nalidixic acid (66.7%), and Gentamicin (11.1%) was the lowest.

Pseudomonas is a food-poisoning microorganism that poses a threat to people's health. This study's goal was to assess the variety and occurrence of Pseudomonas species, with a focus on Pseudomonas aeruginosa, which was found to be contaminating frozen poultry products during storage in some markets. 200 frozen chicken product samples, including 40 of each type (breast, thigh, burger, pane, and kofta), were gathered from various stores in the Faiyoum government, Egypt, throughout 2022. Pseudomonas species were isolated from a total of 39.5% of all samples tested, according to the findings. Bacteriological and biochemical analyses revealed the main isolated pseudomonas spp. to be P. aeruginosa (36.7%), followed by P. fluorescens (30.4%), P. putida (15.2%), and P. diminuta (6.3%). Antibiotic sensitivity affirmed the higher sensitivity of the isolates to various antibacterial drugs utilized in Egypt, comprising Tetracycline, Ampicillin, and Penicillin (100%) and being followed by Sulfamethoxazole (86.2%), Chloramphenicol (62%), and Streptomycin (51.15%). On the other hand, Amikacin (86.2%) and Norfloxacin (74.9%) were found to have the highest sensitivity. P. aeruginosa strains that were analyzed by polymerase chain reaction (PCR) were positive for the 16S rDNA unique to P. aeruginosa and carried the toxR (50%) and exoS (30%) virulence genes. Chicken fillets dipped in lemon juice and pomegranate peel extract (PPE) at 5% for 36 hours decreased the P. aeruginosa count by 62.4% and 56.4%, respectively. In conclusion, the current research confirms the contamination of frozen chicken products by Pseudomonas species. Immersion of chicken fillet in lemon juice (5% w/v) and PPE (5% w/v) can increase its quality and lengthen its shelf life by improving its sensual characteristics and implementing a successful approach for reducing P. aeruginosa in chicken products.

Campylobacter is the most common bacterial cause of gastroenteritis globally. A total of 182 fully identified strains of Campylobacter species (42 C. coli and 140 C. jejuni) collected from 6 broiler farms were subjected to studying the antimicrobial resistance pattern and molecular typing of virulence (cadF, ctdA, dnaJ waaC,iam, and fla) and antimicrobial resistance genes (blaOXA-61, gyrA, tetA, tetO, and IR) with sequencing of gyrA region of one strain of fluroquinolones resistant C. coli and C. jejuni. The identified isolates were highly resistant to erythromycin and sulfamethoxazole-trimethoprim. Furthermore, both meropenem and imipenem were effective against the recovered isolates. The tested C. jejuni and C. coli strains had 100%, 83.3%, and 83.3% prevalence of cadF, ctdA, and dnaJ virulence genes, respectively, while waaC, iam, and fla genes couldn’t be detected. The blaOXA-61 resistance gene was found in all of the Campylobacter spp. examined. Furthermore, gyrA, tetA, tetO, and IR resistance genes were found in 100%, 83.3%, 83.3%, and 66.7% of the retrieved C. jejuni strains, respectively.  Likewise, resistance genes were found in 83.3%, 83.3%, 83.3%, and 66.7% of the retrieved C. coli strains, respectively. Approximately 58% (7/12) of the Campylobacter spp. recovered were MDR. Furthermore, 50% (3/6) of the C. jejuni strains recovered were MDR, while 66.7% (4/6) of the C. coli isolates recovered were MDR with MARI(0.22-0.55). For detection of mutations of the gyrA gene, the sequence data of two isolates (C. jejuni and C.coli) were analyzed against the reference sequence on the gene bank where the C. jejuni strain had six mutations, while the C. coli strain had twenty-three. The current findings suggest that MDR Campylobacter strains in poultry may be able to transmit highly virulent Campylobacter as a foodborne pathogen.