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Objectives: This study aimed to prove the effects of Escherichia coli isolates isolated from diseased broilers to form biofilms, describe their antimicrobial sensetivity, and determine the effect of allicin and cinnamon essential oils on the expression of some genes (fimH, int1, and luxS) through quantitative polymerase chain reaction (q-PCR). Materials and Methods: 140 samples were obtained from diseased broilers in Beni-Suef Governorate, Egypt. These samples were examined by conventional bacteriology methods to detect the causative agent. The antimicrobial susceptibility of the isolated bacteria was assessed using the disc diffusion method, The ability of yeast extract-casamino acids Congo Red Agar to generate phenotypic biofilms was next tested. The presence of resistance and virulence genes in some multidrug resistant isolates was genotypically investigated. The antibacterial effects of allicin and cinnamon oil were evaluated against the growth of multidrug-resistant E. coli. Finally, q-PCR was utilized to assess changes in some genes' expression. Results: Escherichia coli was isolated from 61 samples (43.6%). An antimicrobial susceptibility test revealed that multidrug-resistance (MDR) (could resist more than three antimicrobial classes) E. coli prevalence was 100%. 40.8% of isolates phenotypically produce biofilms. The detection of resistance and virulence genes by PCR showed that all tested isolates carry aadB, fimH, int1, qnrS, and luxS genes, while only 40% harbor iss genes. q-PCR showed that after treatment with allicin and cinnamon oils, gene expression went down. Conclusion: This investigation highlights that E. coli showed resistance against most of the tested antimicrobials; all isolates were MDR. The study showed wide dissemination of virulence and resistance genes among E. coli. Allicin and cinnamon oils have antimicrobial activities and could be used as alternatives to synthetic antimicrobial agents.
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OBJECTIVE: The objectives of this study were to determine the biofilm-forming capability and antimicrobial susceptibility of Escherichia coli recovered from bovine endometritis samples. MATERIALS AND METHODS: A total of 120 uterine specimens were collected from cows suffering from endometritis for bacteriological examination. Antimicrobial susceptibility testing was carried out for all isolated E. coli by using the disc diffusion method. The isolates were phenotypically studied for biofilm-forming ability by cultivation on yeast extract -casamino acids Congo red agar (CRA). Some randomly selected isolates were chosen for the molecular identification of some virulence and resistance genes. RESULTS: A total of 58(48.3%) E. coli isolates could be isolated from the 120 samples. Antimicrobial susceptibility testing exhibited that 91.4%, 79.3%, 79.3%, 74.1%, and 58.6% of the isolates were sensitive to gentamicin, amoxicillin-clavulanic acid, ciprofloxacin, cephalexin, and sulfamethoxazole- trimethoprim, respectively. On the other hand, 91.4% and 70.7% isolates were resistant to cefotaxime and doxycycline, respectively. Cultivation on CRA revealed that 46.6% of isolates were biofilm producers. The molecular detection of resistance and virulence genes declared that all isolates harbored bla TEM, sul1, tetA, qnrS, bla CTX-M , and fimH with a percentage of 100%, papC (40%), and hlyA (10%). FimH was the most prevalent biofilm-associated gene. CONCLUSION: The present study highlights the high prevalence of multi-drug- resistant E. coli associated with bovine endometritis. The detection of the fimH gene is circumstantial evidenced that this gene has a crucial role in biofilm formation in intrauterine pathogenic E. coli.
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OBJECTIVE: This study was performed to probe the antimicrobial resistance and virulence genes profiling in Pseudomonas aeruginosa recovered from the cases of pericarditis in broiler chickens. MATERIALS AND METHODS: The samples (n = 250) collected from the cases of pericarditis in broiler chickens were bacteriologically examined. Antimicrobial susceptibility was tested by disc diffusion technique. The isolates were genotypically studied for the presence of antimicrobial resistance and virulence gene traits. Finally, the nucleotide sequence of representative resistance gene (mexR gene) and virulence genes (toxA and lasI genes) was analyzed. RESULTS: P. aeruginosa was isolated from 45 samples (18%). Antimicrobial susceptibility testing revealed multidrug resistance in most of the recovered P. aeruginosa isolates, whereas colistin and imipenem were the furthermost in vitro-sensitive antibiotics. Antimicrobial resistance genes, such as bla CTX, fox, and mexR, were prevalent in 100%, 80%, and 100% of the isolates, respectively. PCR confirmed virulence genes such as toxA, exoY, lasB, and lasI in 100%, 60%, 80%, and 80% of the isolates, respectively. Nucleotide sequence analysis of representative resistance gene (mexR gene) and virulence genes (toxA and lasI genes) revealed a high correlation between P. aeruginosa recovered from pericarditis in broiler chickens in the present study with PAO1 (reference strain) and with other sequences published on the GenBank representing different localities worldwide. CONCLUSION: It could be concluded that P. aeruginosa recovered from pericarditis in broiler chickens in the current study is highly virulent bacteria, resisting most of the therapeutic agents which not only bear hazards for poultry industry but also represent a public health concern.