This study aimed to investigate the prevalence and antimicrobial resistance of enterococci- and ESBL-producing E. coli isolated from milk of bovine mastitis cases in Egypt. Fifty milk samples of dairy animals were collected from localities in the Nile Delta region of Egypt. Isolates were identified using MALDI-TOF MS, and antibiotic susceptibility testing was performed by the broth microdilution method. PCR amplifications were carried out, targeting resistance-associated genes. Seventeen Enterococcus isolates and eight coliform isolates could be cultivated. Vancomycin resistance rate was high in Ent. faecalis. The VITEK 2 system confirmed all E. coli isolates as ESBL-producing. All Ent. faecalis isolates harbored erm(B), tetL and aac-aphD genes. The vanA gene was detected in Ent. faecalis isolate, vanB was found in other Enterococcus, while one isolate of E. casseliflavus exhibited the vanA gene. E. coli isolates exhibited high prevalence of erm(B) and tetL. E. coli isolates were analyzed by DNA microarray analysis. Four isolates were determined by O-serotyping as O8 (n = 1), O86 (n = 2) and O157 (n = 1). H-serotyping resulted in H11, H12, H21 (two isolates each) and one was of H16 type. Different virulence-associated genes were detected in E. coli isolates including lpfA, astA, celB, cmahemL, intI1 and intI2, and the iroN gene was identified by DNA microarray analysis.
One hundred samples of imported Brazilian frozen meat (50 frozen cubic meat and 50 minced meat) were collected from different supermarkets in El-Menoufia Governorate, Egypt to be examined bacteriologically for detection of Pseudomonas species. The incidence of Pseudomonas species were (35/50) 70% in frozen cubic meat. While, the incidence of such organism in the examined frozen minced meat samples was 80% (40/50). Psychrotrophic bacterial count in the examined frozen cubic meat ranged from 6 x 10 2 to 1.9 x 10 5 with mean value 2.24 x 10 4 cfu/g. In addition, Psychrotrophic bacterial count in frozen minced meat ranged from 7 x 10 2 to 9 x 10 5 with mean value 1.7 x 10 5 cfu/g. The incidence of identified Pseudomonas species (number and percentages) detected in the examined samples of frozen meat represented by Ps. aeruginosa, Ps. fluorescence, Ps. diminuta, Ps putida and Ps. fragi were 15(30%), 40(80%), 8(16%), 5(10%) and 4(8%), respectively. Regarding the minced meat samples the incidence of identified Ps. aeruginosa, Ps. fluorescence, Ps. diminuta, Ps. putida, Ps. fragi were 20(40%), 45(90%), 5(10%), 7(14%) and 8(16%), respectively. The Pseudomonas species were resistant to Oxacillin. They were sensitive to Gentamycin except Ps. fluorescence. PCR is rapid and reliable tool for identification of different bacterial species.
The present study was performed on 250 random samples of fresh meat and meat products. Beef burger, kofta, minced meat and sausage (50 for each) were collected from different shops (25gm of each sample) at Kaliobia Governorate, Egypt, to detect the prevalence of some toxigenic food-borne bacteria, beside the phenotypic characterization and detection of some virulence genes. Bacteriological examination of the collected samples resulted in an isolation of Staph. aureus isolates (41/16.4%), E. coli (25 /10.0%), B. cereus (21/8.4%) and Salmonella (3/1.2%). The antibiotic sensitivity tests for the isolated strains showed multiple antibiotic resistances (ampicillin, methicillin, oxytetracycline, amoxicillin, streptomycin, erythromycin, doxycycline and cefotaxime). Therefore, E. coli; Staph. aureus and B. cereus strains especially antibiotic resistances ones are meat-borne pathogens of public health important Antibiotic resistant Food borne pathogen Meat products
The study was applied on 175 random samples of milk and milk products (white cheese, kareish cheese, yoghurt and ice cream) collected from different shops (35 of each), at Kaliobia governorate, Egypt, for detection of Y. enterocolitica strains, beside the phenotypic characterization and detection of some antibiotic resistant virulence genes in the examined samples. The results revealed that 9(10.9%) Y. enterocolitica isolates, bio types 1A and 1B only were isolated from milk and Kareish cheese samples (5/14.3% for each), (4/11.4%) white cheese, (3/8.6%) ice cream, and (2/5.7%) from yoghurt samples. The antibiotic sensitivity profile showed that, the isolated Y. enterocolitica isolates were very high resistant for Penicillin-G followed by methicillin, ampicillin, oxytetracycline, amoxicillin, ampicillin, streptomycin and erythromycin. Meanwhile, they were highly sensitive to meropenem and norfloxacin followed by gentamycin, ciprofloxacin, cefotaxime and fluorophenol. PCR declared that blaTEM and tetA genes were detected in all eight studied Y. enterocolitica isolates. So, it was concluded that, the presence of antimicrobial resistant Y. enterocolitica strains in dairy products could be a public health concern for the consumers.
Mycoplasma species are the main important causative agents causing pneumonia in cattle. There are about 200 species or more. The current work aimed to investigate various mycoplasma spp. isolated from the respiratory tract of cattle by microbial culture, conventional polymerase chain reaction technique, and target gene sequencing with phylogenetic analysis. An initial screening was done to confirm the presence of mycoplasma spp. by culture on PPLO's agar, digitonin sensitivity and biochemical tests. 129 isolates were characterized by fried egg with depressed center colonies, digitonin sensitive and negative to glucose fermentation and arginine utilization test. Out of 305 samples, 20 samples were selected for amplification by PCR technique using Mycoplasma 16S rRNA primer. Seven samples were positive to Mycoplasma species and gave amplified band at 1013 bp. subsequently, the seven isolates were sequencing. Four sequenced isolates (EGS1, EGL6, EGL1 and EGS2) were closely related to each other and very close to Mycoplasma bovis strains and far from M. leachii 99/014/6. Two sequenced isolates (EGL2 and EGL5) were closely related to each other and very close to Mycoplasma bovirhinis strains and far from M. leachii 99/014/6. One sequenced isolate (EGL3) was very close to Mycoplasma arginini strain and far from M. bovis 99/014/6. From these results, we can conclude that conventional culture methods for diagnosis either by isolation and identification of Mycoplasma is a timeconsuming method to diagnose mycoplasma infection. So, these methods can be replaced by PCR and genome analysis technology.
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