鸽源鼠伤寒沙门菌的分离鉴定及致病性分析

doi:10.11843/j.issn.0366-6964.2023.11.042

Abstract

Abstract: The aim of this study was to investigate the phylogenetic grouping, drug resistance, virulence and resistance gene carriage and pathogenicity of Salmonella spp. carried by diseased pigeons in a region of Guangdong, and to propose a basis for the prevention and control of Salmonella spp. diseases in the region. In this study, 72 samples of diseased pigeon tissues were isolated, purified and identified. We determined the strain type by 16S rRNA sequencing, serotype identification, and PCR identification of Salmonella typhimurium-specific primers, and detected the drug resistance, resistance gene, and virulence genes of isolated bacteria, and carried out animal pathogenicity tests using mice. We found that the morphology, biochemical indexes, 16S rRNA sequence ratio, serotype identification and specific primer PCR results of the 12 isolates were consistent with the characteristics of Salmonella typhimurium; Resistance results showed that 12 strains were resistant to nalidixic acid and rifampicin, and highly sensitive to trimethoprim, co-trimoxazole, enrofloxacin, florfenicol, ampicillin, and meropenem; We found that 15 drug resistance gene test results showed that 7 strains contained tetA, 5 strains contained sul-11, and other resistance genes were not detected. The results of 17 virulence gene tests showed that the virulence gene carrier rate of 5 strains was 100%, and the virulence gene carrier rate of the remaining strains was also high; In addition, we found that in the animal pathogenicity test, the body weight and feed intake of mice in the experimental group decreased significantly, HE staining observed that the number of liver withered cells increased, the number of jejunal goblet cells decreased, and VH/CD decreased. The 12 strains isolated in this study all belonged to Salmonella typhimurium, all of which showed multi-drug resistance and high virulence gene carrier rate. Animal tests have observed that pathogenic bacteria can cause strong inflammatory damage to animal body organs and are highly pathogenic. We recommend the use of highly sensitive drugs such as trimethoprim for the treatment of salmonellosis in pigeons, or treatment with antibiotic substitutes, regular detection of bacterial resistance, and timely adjustment of treatment regimens.

Key words: Salmonella typhimurium, pigeon, resistance genes, virulence genes, pathogenicity

CLC Number:

S852.61

YANG Menglin, ZHENG Shiqi, PENG Kai, WANG Wei, HUANG Yanhua, PENG Jie. Isolation and Identification of Pigeon-derived Salmonella Typhimurium and Pathogenic Analysis[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4880-4888.

References 32

[1]	PORTES A B, RODRIGUES G, LEITAO M P, et al. Global distribution of plasmid-mediated colistin resistance mcr gene in Salmonella:A systematic review[J]. J Appl Microbiol, 2022, 132(2):872-889.
[2]	亓玉卓, 梁琳, 林伟东, 等. 鸽源鼠伤寒沙门氏菌的分离鉴定[J]. 动物医学进展, 2021, 42(2):122-127.QI Y Z, LIANG L, LIN W D, et al. Isolation and identification of pigeon-origin Salmonella typhimurium[J]. Progress in Veterinary Medicine, 2021, 42(2):122-127. (in Chinese)
[3]	谈笑, 王娉, 李睿, 等. 动物源性食品中病原菌的耐药性研究进展[J]. 食品科学, 2017, 38(19):285-293.TAN X, WANG P, LI R, et al. Recent advances in understanding antibiotic resistance of pathogens in animal-derived foods[J]. Food Science, 2017, 38(19):285-293. (in Chinese)
[4]	OGUNREMI D, NADIN-DAVIS S, DUPRAS A A, et al. Evaluation of a multiplex PCR assay for the identification of Salmonella serovars enteritidis and typhimurium using retail and abattoir samples[J]. J Food Prot, 2017, 80(2):295-301.
[5]	赵巧雅, 刘娜, 潘华, 等. 2株鸽源沙门氏菌病原的分离鉴定及诊治分析[J]. 家禽科学, 2020(2):17-21.ZHAO Y Q, LIU N, PAN H, et al. Isolation, identification, diagnosis and treatment of two Salmonella strains isolated from pigeon[J]. Poultry Science, 2020(2):17-21. (in Chinese)
[6]	邓珊珊, 张海, 蔺俐仲, 等. 规模化猪场猪大肠杆菌的耐药性及相关耐药基因检测[J]. 养猪, 2021(1):113-115.DENG S S, ZHANG H, LIN L Z, et al. Drug resistance and related gene retection of escherichia coli in large-scale pig farms[J]. Swine Production, 2021(1):113-115. (in Chinese)
[7]	OLESEN I, HASMAN H, AARESTRUP F M. Prevalence of β-lactamases among ampicillin-resistant Escherichia coli and Salmonella isolated from food animals in Denmark[J]. Microb Drug Resist, 2004, 10(4):334-340.
[8]	ARLET G, ROUVEAU M, PHILIPPON A. Substitution of alanine for aspartate at position 179 in the SHV-6 extended-spectrum β-lactamase[J]. FEMS Microbiol Lett, 1997, 152(1):163-167.
[9]	CAVACO L M, FRIMODT-MØLLER N, HASMAN H, et al. Prevalence of quinolone resistance mechanisms and associations to minimum inhibitory concentrations in quinolone-resistant Escherichia coli isolated from humans and swine in Denmark[J]. Microb Drug Resist, 2008, 14(2):163-169.
[10]	CATTOIR V, WEILL F X, POIREL L, et al. Prevalence of qnr genes in Salmonella in France[J]. J Antimicrob Chemother, 2007, 59(4):751-754.
[11]	WATERS S H, ROGOWSKY P, GRINSTED J, et al. The tetracycline resistance determinants of RP1 and Tn1721:nucleotide sequence analysis[J]. Nucleic Acids Res, 1983, 11(17):6089-6105.
[12]	SENGELØV G, AGERSØ Y, HALLING-SØRENSEN B, et al. Bacterial antibiotic resistance levels in Danish farmland as a result of treatment with pig manure slurry[J]. Environ Int, 2003, 28(7):587-595.
[13]	张启龙, 栗云鹏, 傅彩霞, 等. 1株信鸽源鼠伤寒沙门氏菌的分离鉴定及其耐药性和毒力分析[J]. 中国畜牧兽医, 2021, 48(1):338-347.ZHANG Q L, LI Y P, FU C X, et al. Isolation, identification, drug resistance and virulence analysis of one Salmonella typhimurium strain from racing pigeons[J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(1):338-347. (in Chinese)
[14]	CHEN S, ZHAO S H, WHITE D G, et al. Characterization of multiple-antimicrobial-resistant Salmonella serovars isolated from retail meats[J]. Appl Environ Microbiol, 2004, 70(1):1-7.
[15]	程琼, 庞瑞亮, 王若晨, 等. 不同源沙门氏菌对小鼠致病力的比较与毒力基因检测[J]. 中国人兽共患病学报, 2013, 29(5):460-465.CHENG Q, PANG R L, WANG R C, et al. Comparative study on pathogenicity of Salmonella isolates from differentsources of laboratory mice and the detection of their virulence genes[J]. Chinese Journal of Zoonoses, 2013, 29(5):460-465. (in Chinese)
[16]	HAI D, YIN X P, LU Z X, et al. Occurrence, drug resistance, and virulence genes of Salmonella isolated from chicken and eggs[J]. Food Control, 2020, 113:107109.
[17]	黄秀梅, 张倩, 盖文燕, 等. 山东地区屠宰场猪肉污染沙门氏菌菌株毒力基因筛查与ERIC-PCR分型[J]. 中国人兽共患病学报, 2018, 34(8):697-702.HUANG X M, ZHANG Q, GAI W Y, et al. Genetic screening and ERIC-PCR genotyping of Salmonella from pork in slaughterhouses in Shandong Province, China[J]. Chinese Journal of Zoonoses, 2018, 34(8):697-702. (in Chinese)
[18]	LI Q, YIN J, LI Z, et al. Serotype distribution, antimicrobial susceptibility, antimicrobial resistance genes and virulence genes of Salmonella isolated from a pig slaughterhouse in Yangzhou, China[J]. AMB Express, 2019, 9(1):210.
[19]	JOHNSON T J, NOLAN L K. Pathogenomics of the virulence plasmids of Escherichia coli[J]. Microbiol Mol Biol Rev, 2009, 73(4):750-774.
[20]	张临, 卢芳, 付恒峰, 等. 鼠伤寒沙门菌BaeSR对氟喹诺酮类药物耐药性的调控机制[J]. 畜牧兽医学报, 2022, 53(3):894-903.ZHANG L, LU F, FU H F, et al. Regulation mechanism of BaeSR on fluoroquinolones resistance in Salmonella typhimurium[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3):894-903. (in Chinese)
[21]	张璐, 沈青春, 张纯萍, 等. 全基因组测序技术对沙门氏菌血清型和耐药性的预测能力分析[J]. 微生物学报, 2021, 61(12):4038-4047.ZHANG L, SHEN Q C, ZHANG C P, et al. Predictive analysis of whole genome sequencing for Salmonella serotype and antimicrobial resistance phenotypes[J]. Acta Microbiologica Sinica, 2021, 61(12):4038-4047. (in Chinese)
[22]	ENNE V I, KING A, LIVERMORE D M, et al. Sulfonamide resistance in Haemophilus influenzae mediated by acquisition of sul2 or a short insertion in chromosomal folP[J]. Antimicrob Agents Chemother, 2002, 46(6):1934-1939.
[23]	RÅDSTRÖM P, SWEDBERG G. RSF1010 and a conjugative plasmid contain sulII, one of two known genes for plasmid-borne sulfonamide resistance dihydropteroate synthase[J]. Antimicrob Agents Chemother, 1988, 32(11):1684-1692.
[24]	ENNE V I, LIVERMORE D M, STEPHENS P, et al. Persistence of sulphonamide resistance in Escherichia coli in the UK despite national prescribing restriction[J]. Lancet, 2001, 357(9265):1325-1328.
[25]	CERNY O, HOLDEN D W. Salmonella SPI-2 type III secretion system-dependent inhibition of antigen presentation and T cell function[J]. Immunol Lett, 2019, 215:35-39.
[26]	刘书宏, 吴科敏, 周辰瑜, 等. 食源性沙门氏菌分离株致病性研究和毒力相关基因检测[J]. 食品科学, 2018, 39(17):182-186.LIU S H, WU K M, ZHOU C Y, et al. Foodborn Salmonella isolates:pathogenicity evaluation and detection of virulence genes[J]. Food Science, 2018, 39(17):182-186. (in Chinese)
[27]	FÀBREGA A, VILA J. Salmonella enterica serovar typhimurium skills to succeed in the host:virulence and regulation[J]. Clin Microbiol Rev, 2013, 26(2):308-341.
[28]	IBRAHIM D, SEWID A H, ARISHA A H, et al. Influence of Glycyrrhiza glabra extract on growth, gene expression of gut integrity, and Campylobacter jejuni colonization in broiler chickens[J]. Front Vet Sci, 2020, 7:612063.
[29]	PATRA A K, AMASHEH S, ASCHENBACH J R. Modulation of gastrointestinal barrier and nutrient transport function in farm animals by natural plant bioactive compounds- A comprehensive review[J]. Crit Rev Food Sci Nutr, 2019, 59(20):3237-3266.
[30]	GART E V, SUCHODOLSKI J S, WELSH JR T H, et al. Salmonella typhimurium and multidirectional communication in the gut[J]. Front Microbiol, 2016, 7:1827.
[31]	NOTTI R Q, STEBBINS C E. The structure and function of type III secretion systems[J]. Microbiol Spectr, 2016, 4(1), doi:10. 1128/microbiolspec. VMBF-0004-2015.
[32]	LLORENTE C, SCHNABL B. Fast-track clearance of bacteria from the liver[J]. Cell Host Microbe, 2016, 20(1):1-2.

Isolation and Identification of Pigeon-derived Salmonella Typhimurium and Pathogenic Analysis

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