深圳地区动物肉源沙门菌的分离鉴定及耐药性分析
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摘要
在改进沙门菌分离方法的基础上,分离并鉴定深圳市动物肉源沙门菌,并进行耐药性分析。结果:采集自深圳地区的1 055个动物肉源样品的沙门菌总阳性率为21. 04%,其中:鸡肉(28. 71%)>猪肉(24. 95%)>牛肉(15. 00%)>虾肉(2. 82%)。基于最小抑菌浓度(MIC)测定去除重复的耐药表型,得到370株非重复沙门菌株;其中的75. 2%多重耐药,53株(14. 10%)表现ACSSuT~R耐药表型。经血清型鉴定,得到38种血清型及327株确定血清型的沙门菌,优势血清型包括Derby、1,4,[5],12: i: -、Rissen、Agona和8,20: z23,z4: -。在分离动物肉源沙门菌的过程中,四硫酸盐肉汤(TT)相比于氯化镁孔雀绿肉汤(RV)更有效。本研究表明,深圳地区动物肉源样品中,沙门菌分离率鸡肉和猪肉高于牛肉和虾肉,血清型以Derby、1,4,[5],12: i: -、Rissen、Agona和8,20: z23,z4: -为主,且多重耐药比例较高。
This study was to understand the prevalence and distribution of Salmonella in animal meat in the Shenzhen area currently,so as to provide theoretical basis for controlling the transmission of Salmonella from animal meat. Firstly,we isolated Salmonella from 1055 meat samples of the livestock and poultry,and found that the total positive rate of the samples was 21. 04% ( 222/1055),and the isolation rates for four kinds of samples were 28. 71% ( 60/209) in chicken,24. 95% ( 142/569) in pork,15. 00% ( 15/100) in beef and 2. 82% ( 5/177) in shrimp,respectively. Moreover,84. 68% ( 188/222) of positive samples could be detected through TT enrichment,but 30. 63% ( 68/222) through RV enrichment. Secondly,370 non-repeated strains were obtained in this study via MIC technique. 75. 2% of the strains showed multiple drug resistance (MDR),and a total of 53 ( 14. 10%,53/370) strains showed an ACSSu TRdrug resistance phenotype. The data also showed that 286 (77. 29%) strains could be obtained from the samples enriched through TT broth,but only 84( 22. 70%) from samples through RV. Thirdly,we analyzed 370 non-repeated strains through serotyping technology and found that 38 serotypes could be obtained and 327 strains were identified as specific serotypes. Derby,1,4,[5],12: i:-,Rissen,Agona and 8,20: z23,z4:-were among the most prevalent serotypes and 35 of 38 serotypes could be identified from the TT enriched broth,which was almost two times those from the RV broth (35/19),especially for the major serotypes including Derby,Agona and Rissen. alto sum up,among animal meat samples collected in the Shenzhen area,the isolation rate of Salmonella in chicken meat was the highest,followed by pork,beef and shrimp meat samples,and the most common serotypes contained Derby,1,4,[5],12: i:-,Rissen,Agona and 8,20: z23,z4:-. Higher multiple drug resistance rate of Salmonella was observated,and TT enrichment broth was likely more effective than RV broth in isolating Salmonella from animal meat.
This study was to understand the prevalence and distribution of Salmonella in animal meat in the Shenzhen area currently,so as to provide theoretical basis for controlling the transmission of Salmonella from animal meat. Firstly,we isolated Salmonella from 1055 meat samples of the livestock and poultry,and found that the total positive rate of the samples was 21. 04% ( 222/1055),and the isolation rates for four kinds of samples were 28. 71% ( 60/209) in chicken,24. 95% ( 142/569) in pork,15. 00% ( 15/100) in beef and 2. 82% ( 5/177) in shrimp,respectively. Moreover,84. 68% ( 188/222) of positive samples could be detected through TT enrichment,but 30. 63% ( 68/222) through RV enrichment. Secondly,370 non-repeated strains were obtained in this study via MIC technique. 75. 2% of the strains showed multiple drug resistance (MDR),and a total of 53 ( 14. 10%,53/370) strains showed an ACSSu TRdrug resistance phenotype. The data also showed that 286 (77. 29%) strains could be obtained from the samples enriched through TT broth,but only 84( 22. 70%) from samples through RV. Thirdly,we analyzed 370 non-repeated strains through serotyping technology and found that 38 serotypes could be obtained and 327 strains were identified as specific serotypes. Derby,1,4,[5],12: i:-,Rissen,Agona and 8,20: z23,z4:-were among the most prevalent serotypes and 35 of 38 serotypes could be identified from the TT enriched broth,which was almost two times those from the RV broth (35/19),especially for the major serotypes including Derby,Agona and Rissen. alto sum up,among animal meat samples collected in the Shenzhen area,the isolation rate of Salmonella in chicken meat was the highest,followed by pork,beef and shrimp meat samples,and the most common serotypes contained Derby,1,4,[5],12: i:-,Rissen,Agona and 8,20: z23,z4:-. Higher multiple drug resistance rate of Salmonella was observated,and TT enrichment broth was likely more effective than RV broth in isolating Salmonella from animal meat.
引文
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[17]Zhang J,Yang X,Kuang D,et al.Prevalence of antimicrobial resistance of non-typhoidal Salmonella serovars in retail aquaculture products[J].Int J Food Microbiol,2015,210:47-52.
[18]Yang B,Qu D,Zhang X,et al.Prevalence and characterization of Salmonella serovars in retail meats of marketplace in Shanxi,China[J].Int J Food Microbiol,2010,141(1-2):63-72.
[19]Hammack T S,Amaguana R M,June G A,et al.Relative effectiveness of selenite cystine broth,tetrathionate broth,and RappaportVassiliadis medium for the recovery of Salmonella spp.from foods with a low microbial load[J].J Food Prot,1999,62(1):16-21.
[20]June G A,Sherrod P S,Hammack T S,et al.Relative effectiveness of selenite cystine broth,tetrathionate broth,and rappaport-vassiliadis medium for recovery of Salmonella spp.from raw flesh,highly contaminated foods,and poultry feed:collaborative study[J].JAOAC Int,1996,79(6):1307-1323.
[21]Rhodes P,Quesnel L B.Comparison of Muller-Kauffmann tetrathionate broth with Rappaport-Vassiliadis (RV) medium for the isolation of salmonellas from sewage sludge[J].J Appl Bacteriol,1986,60(2):161-167.
[22]Hopkins K L,Kirchner M,Guerra B,et al.Multiresistant Salmonella enterica serovar 4,[5],12:i:-in Europe:a new pandemic strain?[J].Euro Surveill,2010,15(22):19580.
[23]Yang X,Wu Q,Zhang J,et al.Prevalence and characterization of monophasic Salmonella Serovar 1,4,[5],12:i:-of food origin in China[J].PLoS One,2015,10(9):e0137967.
[24]Kim A,Lim S K,Lee K,et al.Characterization of Salmonella enterica Serovar 4,[5],12:i:-isolates from Korean food animals[J].Foodborne Pathog Dis,2015,12(9):766-769.
[25]Ailes E.The Emergence of Salmonella Serotype I 4,[5],12:i:-in the FoodNet sites,1996-2004[J].Idsa,2005.
[26]Lopes M,Galhardo J A,Oliveira J T D,et al.Pesquisa de Salmonella spp.e microrganismos indicadores em carca9as de frango eágua de tanques de pré-resfriamento em abatedouro de aves[J].Química Nova,2007,28(3):465.
[27]Sousa M ? B,Mendes E N,Penna F J,et al.Acute diarrhea associated with Salmonella enterica in Belo Horizonte-MG:prevalence and characterization of isolates Diarreia aguda associada a Salmonella enterica em Belo Horizonte-MG:prevalência e caracteriza92o das amostras isoladas[J].Jornal Brasileiro De Patologia E Medicina Laboratorial,2013,49(1):34-38.
[28]Ran L,Wu S,Gao Y,et al.Laboratory-based surveillance of nontyphoidal Salmonella infections in China[J].Foodborne Pathog Dis,2011,8(8):921-927.
[29]Liang Z,Ke B,Deng X,et al.Serotypes,seasonal trends,and antibiotic resistance of non-typhoidal Salmonella from human patients in Guangdong Province,China,2009-2012[J].BMC Infect Dis,2015,15:53.
[30]Cray P J,Frye J G,Haro J H,et al.Acssut multi-drug resistance among salmonella isolates of animal origin[C].International Conference on Emerging Antimicrobial Agents in Veterinary Medicine,Prague,Czech Republic,2008.
[2]Jackson B R,Griffin P M,Cole D,et al.Outbreak-associated Salmonella enterica serotypes and food commodities,United States,1998-2008[J].Emerg Infect Dis,2013,19(8):1239-1244.
[3]Kaye K S,Fraimow H S,Abrutyn E.Pathogens resistant to antimicrobial agents.Epidemiology,molecular mechanisms,and clinical management[J].Infect Dis Clin North Am,2000,14(2):293-319.
[4]Arslan S,Eyi A.Occurrence and antimicrobial resistance profiles of Salmonella species in retail meat products[J].J Food Prot,2010,73(9):1613-1617.
[5]Naik V K,Shakya S,Patyal A,et al.Isolation and molecular characterization of Salmonella spp.from chevon and chicken meat collected from different districts of Chhattisgarh,India[J].Vet World,2015,8(6):702-706.
[6]Brands D A,Inman A E,Gerba C P,et al.Prevalence of Salmonella spp.in oysters in the United States[J].Appl Environ Microbiol,2005,71(2):893-897.
[7]Chao G,Zhou X,Jiao X,et al.Prevalence and antimicrobial resistance of foodborne pathogens isolated from food products in China[J].Foodborne Pathog Dis,2007,4(3):277-284.
[8]Gebreyes W A,Thakur S.Multidrug-resistant Salmonella enterica serovar Muenchen from pigs and humans and potential interserovar transfer of antimicrobial resistance[J].Antimicrob Agents Chemother,2005,49(2):503-511.
[9]Yan H,Li L,Alam M J,et al.Prevalence and antimicrobial resistance of Salmonella in retail foods in northern China[J].Int J Food Microbiol,2010,143(3):230-234.
[10]Aslam M,Checkley S,Avery B,et al.Phenotypic and genetic characterization of antimicrobial resistance in Salmonella serovars isolated from retail meats in Alberta,Canada[J].Food Microbiol,2012,32(1):110-117.
[11]Waltman W D,Mallinson E T.Isolation of Salmonella from poultry tissue and environmental samples:a nationwide survey[J].Avian Dis,1995,39(1):45-54.
[12]Bouchrif B,Paglietti B,Murgia M,et al.Prevalence and antibioticresistance of Salmonella isolated from food in Morocco[J].J Infect Dev Ctries,2009,3(1):35-40.
[13]Patel J B.Performance standards for antimicrobial susceptibility testing:twenty-fourth informational supplement[S].Clinical and Laboratory Standards Institute.2014.
[14]Vismanos M F C,Baldrias L R.Occurrence of Salmonella in fresh pork,beef,and poultry in Laguna [Philippines][EB/OL](1998-08)[2018-08-27].http://nco.uplb.edu.ph/xmlui/handle/123456789/272?show=full.
[15]Zhao C,Ge B,De V J,et al.Prevalence of Campylobacter spp.,Escherichia coli,and Salmonella serovars in retail chicken,turkey,pork,and beef from the Greater Washington,D.C.,area[J].Appl Environ Microbiol,2001,67(12):5431-5436.
[16]Abouzeed Y M,Hariharan H,Poppe C,et al.Characterization of Salmonella isolates from beef cattle,broiler chickens and human sources on Prince Edward Island[J].Comp Immunol Microbiol Infect Dis,2000,23(4):253-266.
[17]Zhang J,Yang X,Kuang D,et al.Prevalence of antimicrobial resistance of non-typhoidal Salmonella serovars in retail aquaculture products[J].Int J Food Microbiol,2015,210:47-52.
[18]Yang B,Qu D,Zhang X,et al.Prevalence and characterization of Salmonella serovars in retail meats of marketplace in Shanxi,China[J].Int J Food Microbiol,2010,141(1-2):63-72.
[19]Hammack T S,Amaguana R M,June G A,et al.Relative effectiveness of selenite cystine broth,tetrathionate broth,and RappaportVassiliadis medium for the recovery of Salmonella spp.from foods with a low microbial load[J].J Food Prot,1999,62(1):16-21.
[20]June G A,Sherrod P S,Hammack T S,et al.Relative effectiveness of selenite cystine broth,tetrathionate broth,and rappaport-vassiliadis medium for recovery of Salmonella spp.from raw flesh,highly contaminated foods,and poultry feed:collaborative study[J].JAOAC Int,1996,79(6):1307-1323.
[21]Rhodes P,Quesnel L B.Comparison of Muller-Kauffmann tetrathionate broth with Rappaport-Vassiliadis (RV) medium for the isolation of salmonellas from sewage sludge[J].J Appl Bacteriol,1986,60(2):161-167.
[22]Hopkins K L,Kirchner M,Guerra B,et al.Multiresistant Salmonella enterica serovar 4,[5],12:i:-in Europe:a new pandemic strain?[J].Euro Surveill,2010,15(22):19580.
[23]Yang X,Wu Q,Zhang J,et al.Prevalence and characterization of monophasic Salmonella Serovar 1,4,[5],12:i:-of food origin in China[J].PLoS One,2015,10(9):e0137967.
[24]Kim A,Lim S K,Lee K,et al.Characterization of Salmonella enterica Serovar 4,[5],12:i:-isolates from Korean food animals[J].Foodborne Pathog Dis,2015,12(9):766-769.
[25]Ailes E.The Emergence of Salmonella Serotype I 4,[5],12:i:-in the FoodNet sites,1996-2004[J].Idsa,2005.
[26]Lopes M,Galhardo J A,Oliveira J T D,et al.Pesquisa de Salmonella spp.e microrganismos indicadores em carca9as de frango eágua de tanques de pré-resfriamento em abatedouro de aves[J].Química Nova,2007,28(3):465.
[27]Sousa M ? B,Mendes E N,Penna F J,et al.Acute diarrhea associated with Salmonella enterica in Belo Horizonte-MG:prevalence and characterization of isolates Diarreia aguda associada a Salmonella enterica em Belo Horizonte-MG:prevalência e caracteriza92o das amostras isoladas[J].Jornal Brasileiro De Patologia E Medicina Laboratorial,2013,49(1):34-38.
[28]Ran L,Wu S,Gao Y,et al.Laboratory-based surveillance of nontyphoidal Salmonella infections in China[J].Foodborne Pathog Dis,2011,8(8):921-927.
[29]Liang Z,Ke B,Deng X,et al.Serotypes,seasonal trends,and antibiotic resistance of non-typhoidal Salmonella from human patients in Guangdong Province,China,2009-2012[J].BMC Infect Dis,2015,15:53.
[30]Cray P J,Frye J G,Haro J H,et al.Acssut multi-drug resistance among salmonella isolates of animal origin[C].International Conference on Emerging Antimicrobial Agents in Veterinary Medicine,Prague,Czech Republic,2008.