Molecular Detection of Some Virulence Genes in Pseudomonas aeruginosa Isolated from Chicken Embryos and Broilers with regard to Disinfectant Resistance

Abstract

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.

Phenotypic and Genotypic Characteristics of Antimicrobial and Disinfectant Resistance of Gram-negative Bacteria Involved in Early Broiler Chick Mortality

“…Quaternary Ammonium Compounds also are supposed to destruct the outer membrane of Gram-negative microorganism, consequently enhancing their own uptake (McDonnell and Denver 1999). In our study, qacED1 gene was found in all Gram-negative strains (Salmonella, Escherichia coli, and Pseudomonas aeruginosa) with the same 100% percentage, these results agreed with results reported by various workers (Nabil et al 2019;Shahat et al 2019) as they detected qacED1 gene between E. coli, Salmonella, and P. aeruginosa isolates in 100% case. Between the Gram-negative bacteria, QACs resistance genes are frequently associated with plasmid-mediated class 1 integrons that possess a variety of antimicrobial resistance genes.…”

“…In another report, P. aeruginosa strains isolated from 216 yolk sacs of baby chicks were resistant to florfenicol (El-Sawah et al 2016). All P. aeruginosa isolates recovered from the liver, heart, and yolk sacs of 50 samples of baby chicks were 100% resistant to ampicillin (Shahat et al 2019).…”

“…In Egypt, many studies (EL-Sawah et al 2016;El-Tawab et al 2017;Saad et al 2017;Ibrahim et al 2019) considered E. coli as the most common 2013) find E. coli with the highest percentage followed by S. aureus and P. mirabilis from chicks in Ethiopia. On the other hand, Shahat et al (2019) reported that the incidence of Pseudomonas species in broiler chicks (1-10 days) suffered from diarrhea, yellowish nasal secretion, ruffled feather, and conjunctivitis from different farms was 42%. Single and mixed infections were observed as the most predominant single infections were in E. coli (19%), followed by Salmonella (16%).…”

Evaluation of the Synergistic Effect of Amikacin with Cefotaxime against Pseudomonas aeruginosa and Its Biofilm Genes Expression

“…Exposure to each antibiotic caused a decreased level of biofilm expression ranging between 0.1-and 0.7-fold changes, while the repression was strong and most significant with amikacin-cefotaxime combination treatment with fold change reaching 0.08, i.e., the consequence of treatment on the average expression profile among all biofilm involving genes constituting the bacterial communities studied. As described in this paper and by others [59][60][61], sub-MICs of combinations have potent effects on attenuating biofilm formation which are totally different from each antibiotic alone.…”

Phenotypic characterization of the Egyptian isolates “extensively drug-resistant Pseudomonas aeruginosa” and detection of their metallo-β-lactamases encoding genes

“…According to the results, PCR asserted the presence of P. aeruginosa DNA in all the selected isolates identified conventionally. The confirmation of P. aeruginosa identification via molecular approach turned out to be increasingly imperative to diminish the issues of culture method such as a false negative culture result that may be owing to the sample overgrowth by other bacteria (Shahat et al 2019). Of the acquired metallo-βlactamase genes investigated, NDM-1 had been the most frequently detectable gene among the selected XDR-PA isolates, although the rare occurrence in Egypt.…”