Genotyping of enterotoxigenic methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Staphylococcus aureus (VRSA) among commensal rodents in North Sinai, Egypt.

AIM: This study aimed to identify genotype enterotoxigenic antimicrobial-resistant Staphylococcus aureus species, mainly methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) among commensal rodents. METHODS AND RESULTS: A total of 280 samples were collected from nasal and mouth swabs, heart blood, intestinal content and lung tissues of 56 commensal rodents trapped from North Sinai, Egypt. Antimicrobial susceptibility testing was performed to bacteriologically identified S. aureus isolates against 15 antimicrobial agents by disc diffusion method. Detection was conducted for identifying coagulase gene (coA), antimicrobial-resistant genes (mecA and vanA/B), enterotoxigenic and virulence determinant genes (hlg, seb, sed and see) among the MRSA and VRSA isolates. RESULTS: Staphylococcus aureus species were isolated from 24 (42.86%) out of 56 rodents. Phenotypic examination revealed that all the isolates were multidrug-resistant, whereas two isolates were multiple antibiotic resistant (MAR). Out of 33 examined isolates, 33 (100%) were resistant to oxacillin and amoxicillin, 31 (93.93%) to cefoxitin and 12 (36.36%) to vancomycin. PCR assay revealed that 24 isolates revealed (100%) positivity to coA gene, 17 (70.83%) to mecA gene and 12 (50%) to vanA/B genes. Enterotoxin genes and haemolysin genes were detected among MRSA and VRSA isolates. There was a strong positive correlation between the tested antimicrobial-resistant genes and virulence genes (p > 0.05). CONCLUSIONS: This study demonstrated the occurrence of MRSA and VRSA strains among commensal rodents in North Sinai, Egypt. The detection of enterotoxigenic and virulence genes of the isolated MRSA and VRSA strains indicated the health hazards of food contamination and zoonotic infections. SIGNIFICANCE AND IMPACTS OF THE STUDY: This study emphasizes the role of commensal rodents in maintaining and disseminating multidrug-resistant MRSA and VRSA strains to the environment, animals and human beings.

Prevalence, Antimicrobial Resistance Profiles, Virulence and Enterotoxins-Determinant Genes of MRSA Isolated from Subclinical Bovine Mastitis in Egypt.

Subclinical mastitis caused by Staphylococcus aureus has worldwide public health significance. Here, we aimed to determine the prevalence of S. aureus, antimicrobial resistance profiles, and the virulence and enterotoxins determinant genes of MRSA strains that caused subclinical bovine mastitis. Milk samples were collected from 120 lactating animals (50 buffaloes and 70 dairy cattle) from different farms located in Ismailia Province (Egypt). The collected samples were investigated for subclinical mastitis using a California mastitis test. The total prevalence of S. aureus was 35.9% (84/234) with 36.3% (53/146) in cattle and 31% (31/88) in buffaloes. Antimicrobial susceptibility testing showed that 35.7% (30/84) of the isolated strains were resistant to cefoxitin, defined as methicillin-resistant S. aureus (MRSA), with 37.7% (20/53) in cattle and 32.2% (10/31) in buffaloes. Using PCR, 100% of the tested strains harbored coa and mecA genes, while 86.6% were positive for spa gene, with remarkable gene size polymorphism. Additionally, 10% of the tested strains contained the pvl gene. Further, using multiplex PCR, 26.6% of the tested samples had sea gene, two strains had sec gene and only one strain had sea and sec genes. The seb and sed genes were absent in the tested strains. In conclusion, mecA, coa and spa virulence genes were widely distributed in MRSA strains isolated from bovine milk, whereas the sea gene was the most predominant enterotoxin gene. Notably, this is the first report that emphasizes the prevalence of pvl gene of MRSA isolated from bovine milk in Egypt.

The occurrence of the multidrug resistance (MDR) and the prevalence of virulence genes and QACs resistance genes in E. coli isolated from environmental and avian sources.

Colibacillosis is a major disease affecting poultry leads to high morbidity and mortality which causing tremendous economic losses worldwide. These economic disparities are amplified among low and middle-income where sanitation and hygiene are challenged by the increasing demand for quality sources of animal protein. With a view to investigating the prevalence of virulence genes and QACs resistance genes as well as monitoring the antibiogram of E. coli strains, a total of 368 specimens were collected from diseased broiler chickens (n = 226) and environmental sources (n = 142) at large-scale poultry farms in Ismailia Governorate, Egypt. The bacteriological examination proved that E. coli prevalence was 26.76% and 50.44% in the farm environment and diseased broilers, respectively. In tandem, the isolated E. coli strains were serogrouped, determining the most common serotypes were O78, O1:H7, O91:H21 and O126. Isolates were tested for antimicrobial susceptibility against 12 antibiotics, screened for 4 virulence genes (iss, papC, eaeA, and cfaI), and screened for 3 QACs resistance genes (qacEΔ1, qacA/B, and qacC/D). All the tested strains were positive for iss and papC genes, only 20.3% of the tested strains were positive for eaeA gene, moreover, the examined strains were negative to CFAI gene. Furthermore, all the tested strains were positive for qacEΔ1, qacA/B, and qacC/D genes. In conclusion; virulence genes (iss, papC) as well as QACs resistance genes are common in avian Pathogenic E. coli and environmental strains and are mainly associated with multi-drug resistance phenomena.

Prevalence of antibiotic resistant mastitis pathogens in dairy cows in Egypt and potential biological control agents produced from plant endophytic actinobacteria.

Dairy production is threatened by antibiotic resistant pathogens worldwide, and alternative solutions to treat mastitis are not available. The prevalence of antibiotic resistant strains is not well known in less developed countries. The prevalence of pathogenic bacteria and their resistance to 21 commercial antibiotics were studied in milk samples taken from 122 dairy cows suffering from the symptoms of mastitis in Egypt. The bacterial species were identified with molecular methods, and antibiotic resistance was studied with disc diffusion method. The prevalence of Streptococcus aureus, Escherichia coli and Pseudomonas aeruginosa were 30%, 17% and 3.5%, respectively. Most (90%) of the S. aureus strains showed resistance to penicillin whereas only 10% of the strains were resistant to oxacillin. Nearly half (40%) of E. coli strains showed resistance to streptomycin. Six P. aeruginosa strains showed resistance to several antibiotics, including ceftriaxone, enrofloxacin and levofloxacin. This points out that despite P. aeruginosa was not common, it should be followed up carefully. Potential biocontrol agents against antibiotic resistant mastitis bacteria were searched among 30 endophytic actinobacterial strains derived from wild medicinal plants. Three plants, namely Mentha longifolia, Malva parviflora and Pulicaria undulata were chosen for a more detailed study; their endophytic actinobacteria were used to prepare metabolic extracts. The crude metabolites of the actinobacteria were extracted with ethyl acetate. All metabolic extracts inhibited the growth of S. aureus, methicillin-resistant Staphylococcus aureus (MRSA), E. coli and P. aeruginosa in vitro. The 16S rRNA sequence analysis revealed that the most efficient actinobacterial strains were two Micromonospora sp. and one Actinobacteria bacterium. We conclude that the combination of the metabolites of several endophytic actinobacteria derived from several medicinal plants would be the most efficient against pathogens. Different metabolite cocktails should be studied further in order to develop novel biocontrol agents to treat antibiotic resistant mastitis bacteria in dairy cows.