山东省鸡源沙门氏菌的分离鉴定及毒力基因分析

doi:10.16431/j.cnki.1671-7236.2021.08.040

Abstract

Abstract: This experiment was aimed to investigate the prevalence and virulence genes distribution of Salmonella from chicken in Shandong province.Samples were collected from large-scale chicken farms in Shandong province, and tetrathionate broths (TTB) were used as Salmonella selective enrichment broths before bacterial isolation.The isolated strains were purified and cultured, and Gram staining, biochemical tests, colony morphology, PCR amplification with Salmonella and serotype specific primers were carried out.33 kinds virulence genes were detected by PCR to study the virulence genes distribution.The results showed that the isolated strains had grown on LB and MacConkey media with colorless, translucent, smooth and neat edged colonies, and the bacterial morphology were Gram-negative brevibacterium with blunt ends at both ends.25 Salmonella strains were isolated, including 9 Salmonella Enteritis, 8 Salmonella Pullorum strains and 8 Salmonella Typhimurium strains.The detection rates of 20 genes that virulence island (invJ, virK, sopA, mogA, hilA, hisJ, ssaB, ssaQ, ssiD, misL, mgtC, orf319, siiE, siiD, bcfA, pipC, sopB, araB, spoB and avrA genes), enterotoxin gene (stn gene) and fimbriae virulence genes (fimA gene) were 100%.The detection rates of 2 virulence island genes (sipA and sodC genes) and 4 virulence plasmid genes (spvA, spvC, spvD and spvR genes) were 96%.The detection rate of spoE gene was 68%, fliC gene was 36%, gipA and spvB genes were 32%, and sseL gene was 4%.In the 25 Salmonella strains, the numbers of carrying virulence genes were 31(8 strains), 30(1 strain), 29(15 strains) and 24(1 strain), respectively.gipA and spvB genes could be detected only in Salmonella Typhimurium, and the positive rate was 100%.The above results showed that Salmonella Enteritis, Salmonella Pullorum and Salmonella Typhimurium were the main pathogens of salmonellosis in Shandong province.All the chicken Salmonella carried many virulence genes, gipA and spvB genes detected only in Salmonella Typhimurium, it could be used as one of the candidate genes for the identification of Salmonella Typhimurium.

Key words: chicken; Salmonella; isolation; identification; virulence gene

CLC Number:

S852.61⁺2

YANG Wenwen, LI Yubao, LU Jianbiao, SI Zhenshu, ZHANG Kaiyue, GUAN Yutang, PANG Zheyu, XU Xinlu, MENG Fanda, DING Ningning. Isolation,Identification and Virulence Gene Analysis of Salmonella from Chickens in Shandong Province[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 3069-3078.

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Isolation,Identification and Virulence Gene Analysis of Salmonella from Chickens in Shandong Province

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