Pseudomonas is a communal motif of environmental associated disease and causes a serious problem in poultry farms, so this study was deliberated to investigate the quandary of Pseudomonas species especially Pseudomonas aeruginosa (P. aeruginosa) which has multifarious virulence genes and plays a major role in poultry outbreaks. Also, it focuses the light on the problem of antimicrobial and disinfectant resistance. A total of 200 samples (100 from dead in shell chicken embryos and 100 from broilers at different ages) were collected from different hatcheries and farms in Luxor governorate, 40 isolates (20%) of Pseudomonas species were isolated and identified serologically as P.cepacia, P.fluorescens, P.putida, P.fragi and P.aeruginosa. PCR inveterate the existence of P.aeruginosa DNA in seven isolates by using 16SrDNA primers at 956bp. P.aeruginosa isolates have different virulence genes such as toxA, exoS, lasB, lasI and oprL gene with incidence rate 71.42% for each of them, except oprL was 100%. Also, Quaternary Ammonium Compounds resistant genes (QACs) were detected in P. aeruginosa isolates with incidence rate (14.28%) for each of qacAB and qacCD genes, while the qacED1 gene incidence was (100%). P. aeruginosa isolates showed an obstacle of antimicrobial resistance for different antimicrobials while most of these isolates cleared susceptibility for ciprofloxacin and norfloxacin. In conclusion, this work described the problem of P. aeruginosa as it proved a high virulence repertoire owned by the P. aeruginosa that confirming its pathogenicity for chicken embryos and broilers. Also, our study is fuelling the concern on disinfectant resistance problem and displaying the relation between QACs and antibiotic resistance. So, the deterrence of the Pseudomonas infection in the poultry housing becomes necessary by applying strict bio-security measures.
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.
In the last 40 years, novel viruses have evolved at a much faster pace than other pathogens. Viral diseases pose a significant threat to public health around the world. Bovines have a longstanding history of significant contributions to human nutrition, agricultural, industrial purposes, medical research, drug and vaccine development, and livelihood. The life cycle, genomic structures, viral proteins, and pathophysiology of bovine viruses studied in vitro paved the way for understanding the human counterparts. Calf model has been used for testing vaccines against RSV, papillomavirus vaccines and anti-HCV agents were principally developed after using the BPV and BVDV model, respectively. Some bovine viruses-based vaccines (BPIV-3 and bovine rotaviruses) were successfully developed, clinically tried, and commercially produced. Cows, immunized with HIV envelope glycoprotein, produced effective broadly neutralizing antibodies in their serum and colostrum against HIV. Here, we have summarized a few examples of human viral infections for which the use of bovines has contributed to the acquisition of new knowledge to improve human health against viral infections covering the convergence between some human and bovine viruses and using bovines as disease models. Additionally, the production of vaccines and drugs, bovine-based products were covered, and the precautions in dealing with bovines and bovine-based materials.
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.
The aim of this study was to investigate the dominant species of Aeromonas in naturally infected African Sharptooth catfish, Clarias gariepinus, in Qena, Egypt and the distribution of two virulence genes among the isolates to assess their pathogenicity. Twenty-five isolates of Aeromonas were recovered from infected fish showing signs of septicemia. Restriction-fragment-length-polymorphism (RFLP) analysis of the 16S-rDNA amplified products demonstrated that the specie isolated were Aeromonas hydrophila (56%) and Aeromonas veronii (44%). Isolates were screened for the cytotoxic enterotoxin, act, and aerolysin, aerA, genes. The act gene was detected only in A. hydrophila, while the aerA gene was more frequently found among all isolates. Catfish challenged with an A. hydrophila isolate that have both the act and aerA genes showed higher mortalities (80.9%) and more severe signs of septicemia than those challenged with an isolate that lacks both genes studied.
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 (blaCMY, blaMIR, 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.
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.
Acinetobacter baumannii (A. baumannii) is an opportunistic pathogen associated with nosocomial infections. In this study, 100 raw milk samples were collected from Qena, Egypt, and subjected to conventional and molecular assays to determine the presence of A. baumannii and investigate their antimicrobial resistance and biofilm formation. Our findings revealed that, among the 100 samples, Acinetobacter spp. were found in 13 samples based on CHROM agar results. We further characterized them using rpoB and 16S-23SrRNA sequencing and gyrB multiplex PCR analysis and confirmed that 9 out of the 13 Acinetobacter spp. isolates were A. baumannii and 4 were other species. The A. baumannii isolates were resistant to β-lactam drugs, including cefotaxime (44%), ampicillin-sulbactam and levofloxacin (33.3% for each), imipenem, meropenem and aztreonam (22.2% for each). We observed different antimicrobial resistance patterns, with a multi-antibiotic resistant (MAR) index ranging from 0.2 to 0.3. According to the PCR results, blaOXA-51 and blaOXA-23 genes were amplified in 100% and 55.5% of the A. baumannii isolates, respectively, while the blaOXA-58 gene was not amplified. Furthermore, the metallo-β-lactamases (MBL) genes blaIMP and blaNDM were found in 11.1% and 22.2% of isolates, respectively, while blaVIM was not amplified. Additionally, eight A. baumannii isolates (88.8%) produced black-colored colonies on Congo red agar, demonstrating their biofilm production capacity. These results showed that, besides other foodborne pathogens, raw milk should also be examined for A. baumannii, which could be a public health concern.
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