Background and Aim: Raw milk is considered an essential source of nutrition during all stages of human life because it offers a valuable supply of protein and minerals. Importantly, milk is considered a good media for the growth and contamination of many pathogenic bacteria, especially food-borne pathogens such as Escherichia coli. Thus, the objective of this study was to characterize E. coli and detect its virulence factors and antibiotic resistance from raw milk samples. Materials and Methods: Raw milk samples (n=100) were collected from different localities in Qena, Egypt, and investigated for the presence of E. coli using different biochemical tests, IMViC tests, serotyping to detect somatic antigen type, and molecularly by polymerase chain reaction (PCR) tests. The presence of different virulence and antimicrobial genes (hly, eae, stx1, stx2, blaTEM, tetA(A), and tetB genes) in E. coli isolates was evaluated using PCR. Results: The results demonstrated that 10 out of 100 milk samples were contaminated with E. coli. Depending on serology, the isolates were classified as O114 (one isolate), O27 (two isolates), O111 (one isolate), O125 (two isolates), and untypeable (five isolates) E. coli. The sequencing of partially amplified 16S rRNA of the untypeable isolates resulted in one isolate, which was initially misidentified as untypeable E. coli but later proved as Enterobacter hormaechei. Moreover, antibacterial susceptibility analysis revealed that nearly all isolates were resistant to more than 3 families of antibiotics, particularly to β-lactams, clindamycin, and rifampin. PCR results demonstrated that all E. coli isolates showed an accurate amplicon for the blaTEM and tetA(A) genes, four isolates harbored eae gene, other four harbored tetB gene, and only one isolate exhibited a positive stx2 gene. Conclusion: Our study explored vital methods for identifying E. coli as a harmful pathogen of raw milk using 16S rRNA sequencing, phylogenetic analysis, and detection of virulence factors and antibiotic-resistant genes.

Bacteria of the genus Pseudomonas are pathogens in both humans and animals. The most prevalent nosocomial pathogen is P. aeruginosa, particularly strains with elevated antibiotic resistance. In this study, a total of eighteen previously identified Pseudomonas species strains, were isolated from chicken. These strains were screened for biofilm formation and antibiotic resistance. In addition, we evaluated clove oil’s effectiveness against Pseudomonas isolates as an antibiofilm agent. The results showed that Pseudomonas species isolates were resistant to most antibiotics tested, particularly those from the β-lactamase family. A significant correlation (p < 0.05) between the development of multidrug-resistant isolates and biofilms is too informal. After amplifying the AmpC-plasmid-mediated genes (bla_CMY, bla_MIR, DHA, and FOX) and biofilm-related genes (psld, rhlA, and pelA) in most of our isolates, PCR confirmed this relationship. Clove oil has a potent antibiofilm effect against Pseudomonas isolates, and may provide a treatment for bacteria that form biofilms and are resistant to antimicrobials.

Introduction: Listeria monocytogenes is an important foodborne pathogen of public- and animal-health concern globally. The persistence of L. monocytogenes in the dairy-processing environment has multifactorial causes, including lack of hygiene, inefficient cleaning, and improper disinfection practices. Materials and Methods: A total of 300 dairy-product and environmental samples were collected from dairy-cattle facilities and local dairy shops and vendors in Qena, Egypt. Samples were screened for the incidence of Listeria spp. and to detect virulence determinants and disinfectant-resistance genes. Three marine algal species — Caulerpa racemosa, Jania rubens, and Padina pavonica — were collected from Hurghada on the Red Sea coast. Algal extracts were screened using gas chromatography–mass spectrometry. The antimicrobial activity of some marine algal extracts, nanoparticles derived therefrom, and some disinfectants against L. monocytogenes strains were assessed in vitro using agar-well diffusion and liquid-broth methods. The impact of P. pavonica extract on the growth and survival of virulent L. monocytogenes in cheese and whey were clarified. Results and Discussion: The incidence of L. monocytogenes in dairy products and environmental samples was 15.5% and 19%, respectively. The most common toxigenic gene profile found among the isolates was hlyA⁺–inlA⁺–prfA⁺. The sensitivity pattern of L. monocytogenes strains to disinfectant containing alkyl (C_{12– 16}) dimethyl BAC was high compared to other tested quaternary ammonium compounds (QAC) disinfectants tested, which showed lower log reductions against resistant strains. The QAC disinfectant–resistance gene qacH was detected in 40% of the isolates. Potent bactericidal activity of a petroleum ether extract of P. pavonica and silver nanoparticles of P. pavonica were obtained against the virulent L. monocytogenes strain. The population of L. monocytogenes in cheese curd and whey after 14 days was reduced at a rate of 9 log CFU/g and 8 log CFU/mL, respectively due to the effect of P. pavonica extract. After 28 days of storage, L. monocytogenes was completely inactivated in those dairy products. Conclusion: P. pavonica extract showed promising antimicrobial properties, calling for further comprehensive studies prior to it being applied in the food industry to enhance the safety, quality, and shelf life of products and protect public health.