This study is designed to discuss the antimicrobial resistance, virulence determinants and biofilm formation capacity of Enterococcus spp. isolated from milk of sheep and goat with subclinical mastitis in Qena, Egypt. The obtained isolates were identified by the VITEK2 system and 16S rDNA sequencing as E. faecalis, E. faecium, E. casseliflavus and E. hirae. Overall, E. faecalis and E. faecium were the dominant species recovered from mastitic milk samples. The antimicrobial susceptibility test evidenced multidrug resistance of the isolates against the following antimicrobials: oxacillin (89.2.%), followed by vancomycin (75.7%) and linezolid (70.3%). Also, most of these isolates (73%) could form biofilms. For example, 18.9% of Enterococcus strains formed strong biofilm, whereas 32.4% of isolates formed moderate biofilm and 21.6% of isolates formed weak biofilm. The most prevalent resistance genes found in our isolates were blaZ (54%), vanA (40%), ermB (51.4%), tetM (13.5%) and optrA (10.8%). Moreover, asa1 (37.8%), cylA (42.3%), gelE (78.4%), esp (32.4%), EF3314(48.6%) and ace (75.5%) were the most common virulence genes. A significant correlation was found between biofilm formation, multidrug resistance and virulence genes of the isolates. This study highlights several aspects of virulence and harmfulness of Enterococcus strains isolated from subclinical mastitic milk, which necessitates continuous inspection and monitoring of dairy animals.

Aim:This study was accomplished to test raw milk and certain dairy products sold in local markets of Qena, Egypt, for the presence of Campylobacter coli and Campylobacter jejuni.Materials and Methods:A total of 150 samples of raw milk, kareish cheese, and yoghurt (50 samples each) were subjected first to enrichment in Bolton broth at 42°C for 2 days under a microaerobic condition, subsequently campylobacter blood free selective agar plates were cultured and incubated in the same condition of the broth. Based on the morphological and biochemical themes of the growing colonies, it was further classified into Campylobacter spp. The identified isolates were later affirmed by polymerase chain reaction using primers that were designed to locate hipO genes in C. jejuni and glyA in C. coli.Results:Of the total 150 examined samples of raw milk and soft cheese samples; 37 (24.6%) samples were contaminated with Campylobacter spp. C. jejuni was dominating in this study in 20%, 14%, and 8% of the examined raw milk, kareish cheese, and yoghurt samples, respectively. No sample harbored C. coli.Conclusion:Campylobacter spp. could be detected in 24.6% of the investigated samples. C. jejuni isolated from 14% of the total tested samples, while C. coli could not be detected from the examined samples. Campylobacter spp. is rampant in the areas of poor hygienic conditions making products made from raw milk of public health hazard.

The objective of this study was to investigate the antimicrobial activity of ethanol extract of propolis against enterotoxigenic strain of MRSA which inoculated into lab prepared ice cream. EEP was added to ice cream in 3 concentrations (150, 300 and 600 mg/L). The prepared ice cream was divided into 2 groups, one stored at freezer temp. at (-5°C), while the other was kept in deep freezer temp. at (-20°C). MRSA could not be counted from the th nd st 4 , 2 and 1 week of storage at freezer temp, while at deep freezer temp. MRSA could not be enumerated from rd st rd the 3 , 1 week and 3 day of storage in portions contained 150, 300 and 600mg/L EEP, respectively.

Objective:The aim of the present work was to investigate the mutual role that may be played by the served dairy food and food handlers in the transmission of methicillin- and vancomycin-resistant Staphylococcus aureus and coagulase-negative Staphylococci to patients who were hospitalized in Qena City, Egypt.Materials and Methods:A total of 210 samples including 90 dairy food samples which offered to the patients in the hospital, 60 nasal and hand swabs from food handlers working in the hospital, and 60 nasal and diarrheal swabs from patients suffering from diarrhea were investigated for the presence of coagulase-positive S. aureus and coagulase-negative Staphylococci, then isolates were screened for methicillin and vancomycin resistance phenotypically and genotypically. 16s rRNA gene sequencing was employed to construct the neighbor-joining tree.Results:Unlike food samples, both coagulase-positive S. aureus and coagulase-negative Staphylococci occurred in human samples. Methicillin- and vancomycin-resistant coagulase-negative Staphylococci could be detected in 41.7% & 20.8%, 68% & 31.9%, and 81.3% & 55.2% of isolates obtained from dairy food, food handlers, and patients’ samples, respectively. Whereas 81% & 64.3%, and 75.4% & 38.6% of coagulase-positive S. aureus obtained from food handlers and patients’ samples exhibited resistance to methicillin and vancomycin, respectively. Phenotypic resistance was confirmed molecularly through detection of mecA and vanA genes.Conclusion:A significant role can be played by food and food handlers in the transmission of methicillin- and vancomycin-resistant Staphylococci to patients, which has been proved in this study through the close phylogenetic relation between S. epidermidis isolated from food, food handlers, and patients’ diarrheal samples.

A total of 60 powdered milk infant formula and milk powder samples were obtained from different localities in Qena Governorate, Egypt, to be evaluated for presence of Enterobacter and closely related species using FDA procedure. Ten out of 60 samples were contaminated with Enterobacter spp. and other Enterobacteriaceae. The obtained isolates were biochemically identified as Enterobacter spp., Klebsiella pneumoniae, Escherichia coli, Citrobacter spp. Further characterization was performed using 16S rRNA sequence analysis; it was a pivotal to confirm the identity of the isolates especially in imprecise identification of phenotypic method in some isolates like Enterobacter hormaechei, Klebsiella quasipneumoniae, and Acinetobacter vivianii. Results revealed the contamination of powdered milk infant formula with E. hormaechei which makes it a matter of concern. According to the best of our knowledge, Enterobacter hormaechei and Klebsiella quasipneumonia were isolated for the first time in Egypt. The phylogenetic analysis proved the close relation between our isolates in amplification of partial 16S rRNA gene. Practical applications As many neonates depend on powdered infant formula in their feeding as well as milk powder is a fundamental component of many infants diet, our study evaluated these products for the presence of pathogens of safety concern. This study focused on Enterobacteriaceae family members other than Salmonella and Cronobacter sakazakii using 16S rRNA gene for identification and differentiation of the closely related species.