J亚群禽白血病病毒与粪肠球菌混合感染的诊断及相关基因分析

doi:10.16431/j.cnki.1671-7236.2021.12.035

Abstract

Abstract: In order to understand the variation of gp85 gene of Avian leukemia virus (ALV) subgroup J and the carrying of virulence gene of Enterococcus faecalis (EF) in a farm in Guiyang city, etiological investigation and analysis were carried out on the cases of co-infection of ALV and EF in this farm.Samples were collected for virus nucleic acid detection, gp85 gene cloning and sequencing analysis, nucleotide sequence similarity analysis, bacterial isolation and identification, virulence gene detection, etc.The results showed that one ALV strain and one Enterococcus faecalis strain were successfully isolated and identified, which were named GZM52021 and GZEF2021 respectively.The specific amplification band of ALV only appeared at about 422 bp, indicating the existence of ALV-J infection.ALV gp85 gene was successfully cloned, and the similarity between GZM52021 strain and gp85 gene of subgroup J reference strain (GenBank accession No.:KM655820, KF796654, JN378888 and Z46390) was high, ranging from 97.9% to 98.2%.The construction results of genetic evolutionary tree of nucleotide sequence of gp85 gene were consistent with the similarity results.GZEF2021 strain grew round convex and opaque milky white colonies on LB solid plate, and there was obvious hemolysis on blood agar plate.Gram staining microscopy showed spherical or oval positive Streptococcus.16S rRNA sequencing showed that the similarity between GZEF2021 strain and Enterococcus faecalis (GenBank accession No.:KJ626240, MT611693, KY399248, MH919370, MN379663 and KU937389) was more than 99%.The results of drug sensitivity test showed that GZEF2021 strain was sensitive to cefoperazone, penicillin, piperacillin and carbenicillin, moderately sensitive to compound sulfamethoxazole, cefradine and cefazolin and resistant to neomycin, amikacin, doxycycline and minocycline.The detection of virulence genes showed that GZEF2021 strain carried ace, gelE, asa1, EF3314 and efaA five virulence genes, indicating that the Enterococcus faecalis isolated from chicken might be pathogenic.The co-infection of ALV subgroup J and Enterococcus faecalis existed in the dead chicken.The results of this experiment could provide some references for understanding the epidemic trend of ALV subgroup J and Enterococcus faecalis and the diagnosis and treatment of mixed infection in Guizhou.

Key words: Avian leukosis virus (ALV); Enterococcus faecalis; mixed infection; virulence gene

CLC Number:

S852.65⁺7

LIU Manman, WEN Guilan, WEN Ming, ZHANG Xiaobo, HU Xuan, SUN Yun, ZHANG Xiuli. Diagnosis and Related Gene Analysis of Co-infection of Avian Leukemia Virus Subgroup J and Enterococcus faecalis[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4671-4680.

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Diagnosis and Related Gene Analysis of Co-infection of Avian Leukemia Virus Subgroup J and Enterococcus faecalis

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