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. 2020 Aug 5;7(3):452-463.
doi: 10.5455/javar.2020.g441. eCollection 2020 Sep.

Virulence and resistance determinants in Pseudomonas aeruginosa isolated from pericarditis in diseased broiler chickens in Egypt

Walid Hamdy Hassan  1 , Ahmed Mohamed Kamel Ibrahim  2 , Salama Abohamra Sayed Shany  3 , Hala Sayed Hassan Salam  1
Affiliations

Virulence and resistance determinants in Pseudomonas aeruginosa isolated from pericarditis in diseased broiler chickens in Egypt

Walid Hamdy Hassan et al. J Adv Vet Anim Res. .

Abstract

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.

Keywords: MDR; P. aeruginosa; PCR; Resistance; Sequence; Virulence.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1.
Figure 1.. PCR amplification of the blaCTX gene at 593-bp. Lane (1–10): Showed +Ve isolates. M = DNA ladder; Pos = Control positive; Neg = Control negative.
Figure 2.
Figure 2.. PCR amplification of the fox gene at 190-bp. Lane(2,3,5,6,7,8,9,10): Showed +Ve isolates, Lane (1,4): Showd -Ve isolates, M = DNA ladder; Pos = Control positive; Neg = Control negative.
Figure 3.
Figure 3.. PCR amplification of the mexR gene at 637-bp. Lane (1–10): Showed +Ve isolates. M = 100 bp DNA ladder; Pos = Control positive; Neg = Control negative.
Figure 4.
Figure 4.. PCR amplification of toxA gene of 396pb fragment. Lane (1–10): Showed +Ve isolates. M = 100 bp DNA ladder; Pos = Control positive; Neg = Control negative.
Figure 5.
Figure 5.. PCR amplification of exoY gene of 289bp fragment. Lane (1,3,4,6,7,8): Showed +Ve isolates, Lane (2,5,9,10): Showed -Ve isolates, M = 100 bp DNA ladder; Pos = Control positive; Neg = Control negative.
Figure 6.
Figure 6.. PCR amplification of lasB gene of 1220 bp fragment, Lane (1,2,3,4,5,6,8,9): Showed +Ve isolates, Lane (7,10) : Showed -Ve isolates, M = DNA ladder; Pos = Control positive; Neg = Control negative.
Figure 7.
Figure 7.. PCR amplification of lasIgene of 606bp fragment. Lane (1,2,3,4,5,6,7,9): Showed +Ve isolates, Lane (8,10): Showed -Ve isolates, M = 100 bp DNA ladder; Pos = Control positive; Neg = Control negative.
Figure 8.
Figure 8.. Phylogenetic analysis of P. aeuroginosa toxA gene. Maximum likelihood tree of 1,000 bootstrap value was conducted after the Clustal W alignment algorithm using MEGA X program. Strain presented in this study is labeled with red triangle while other Egyptian strains are marked with blue square.
Figure 9.
Figure 9.. Phylogenetic analysis of P. aeuroginosa mexR gene. The maximum likelihood tree of 1,000 bootstrap value was conducted after the Clustal W alignment algorithm using MEGA X program. Strain presented in this study is labeled with a red triangle, while other Egyptian strains are labeled with blue square.
Figure 10.
Figure 10.. Phylogenetic analysis of P. aeuroginosa lasI gene. The maximum likelihood tree of 1,000 bootstrap value was conducted after the Clustal W alignment algorithm using the MEGA X program. Strain presented in this study is labeled with a red triangle while other Egyptian strains are labeled with blue square.
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