家禽定点屠宰场不同屠宰区域空气的微生物结构
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摘要
为了解家禽屠宰场空气中微生物分布情况及其结构特点,在浙江选择一家禽定点屠宰场,以菌落总数为指标,比较分析屠宰前和屠宰后各屠宰区域空气沉降微生物的数量,并提取微生物DNA作为模板,经聚合酶链式反应扩增细菌16S rRNA基因的V3～V4区,利用高通量测序技术分析其菌群结构。结果发现家禽屠宰场的不同屠宰区域空气沉降微生物数量有所差异,同时屠宰后微生物数量比屠宰前有所增加,特别是在屠宰后挂禽间菌落总数比屠宰前高出71%。高通量测序显示屠宰前和屠宰后不同屠宰区域空气沉降菌多样性、菌群结构、相对丰度等方面均无显著性差异(P>0.05),主要分布于6个门,优势菌门为厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)和放线菌门(Actinobacteria),总的相对丰度在84%以上。在属的水平上,葡萄球菌属(Staphylococcus)和不动细菌属(Acinetobacter)为优势菌属,分别占36.90%和13%(在前10个属中),此外,在前35个属中,葡萄球菌属(Staphylococcus)、寡养单胞菌属(Stenotrophomonas)和埃希菌属(Escherichia)等条件致病菌属分别占33.75%、4.65%和0.90%。由此说明屠宰场不同屠宰区域沉降菌数量存在一定差异,各屠宰区域均分布有一定数量的条件致病菌,研究为鸡肉微生物污染的溯源和完善家禽屠宰场卫生管理提供科学依据。
In order to understand the distribution and structure of airborne microbial communities in poultry slaughterhouses, the quantity of airborne microorganisms in different areas of a slaughterhouse in Zhejiang province before and after slaughter was compared by measuring the total number of colonies, and total bacterial genomic DNA was extracted from the air samples and used as a template to amplify and sequence the V3–V4 region of the 16 S rRNA gene by high throughout sequencing. The results showed that there was a difference in the number of airborne microorganisms in different slaughter areas of the poultry slaughterhouse and the microbial quantity after slaughter was higher than before slaughter. Especially, the total number of colonies after slaughter was 71% higher than before slaughter in the poultry hanging area. High throughput sequencing showed no significant differences in the diversity, structure and abundance of bacterial communities before and after slaughter for all slaughter areas(P > 0.05). The majority of the identified bacteria belonged to 6 phyla, and the dominant bacteria were Firmicutes, Proteobacteria and Actinobacteria, with total relative abundance of more than 84%. In the top 10 genera, Staphylococcus and Acinetobacter were the dominant bacteria, accounting for 36.90% and 13%, respectively. In addition, Staphylococcus, Stenotrophomonas and Escherichia as conditional pathogenic bacteria accounted for 33.75%, 4.65% and 0.90% in the top 35 genera, respectively. These data suggested that there were some differences in the number of airborne bacteria in different slaughter areas, and a certain number of conditional pathogens were detected in air samples from each area. In conclusion, this study provides a scientific basis for the traceability of microbial contaminants in chicken and improving poultry slaughterhouse hygiene management.
In order to understand the distribution and structure of airborne microbial communities in poultry slaughterhouses, the quantity of airborne microorganisms in different areas of a slaughterhouse in Zhejiang province before and after slaughter was compared by measuring the total number of colonies, and total bacterial genomic DNA was extracted from the air samples and used as a template to amplify and sequence the V3–V4 region of the 16 S rRNA gene by high throughout sequencing. The results showed that there was a difference in the number of airborne microorganisms in different slaughter areas of the poultry slaughterhouse and the microbial quantity after slaughter was higher than before slaughter. Especially, the total number of colonies after slaughter was 71% higher than before slaughter in the poultry hanging area. High throughput sequencing showed no significant differences in the diversity, structure and abundance of bacterial communities before and after slaughter for all slaughter areas(P > 0.05). The majority of the identified bacteria belonged to 6 phyla, and the dominant bacteria were Firmicutes, Proteobacteria and Actinobacteria, with total relative abundance of more than 84%. In the top 10 genera, Staphylococcus and Acinetobacter were the dominant bacteria, accounting for 36.90% and 13%, respectively. In addition, Staphylococcus, Stenotrophomonas and Escherichia as conditional pathogenic bacteria accounted for 33.75%, 4.65% and 0.90% in the top 35 genera, respectively. These data suggested that there were some differences in the number of airborne bacteria in different slaughter areas, and a certain number of conditional pathogens were detected in air samples from each area. In conclusion, this study provides a scientific basis for the traceability of microbial contaminants in chicken and improving poultry slaughterhouse hygiene management.
引文
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[22]WAKINO S,IMAI E,YOSHIOKA K,et al.Clinical importance of Stenotrophomonas maltophilia nosocomial pneumonia due to its high mortality in hemodialysis patients[J].Therapeutic Apheresis&Dialysis,2009,13(3):193-198.DOI:10.1111/j.1744-9987.2009.00693.
[23]BROOKE J S.Stenotrophomonas maltophilia:an emerging global opportunistic pathogen[J].Clinical Microbiology Reviews,2012,25(1):2-41.
[24]JANG T N,WANG F D,WANG L S,et al.Xanthomonas maltophilia bacteremia:an analysis of 32 cases[J].Journal of the Formosan Medical Association,1992,91(12):1170-1176.
[25]VICTOR M A,ARPI M,BRUUN B,et al.Xanthomonas maltophilia bacteremia in immunocompromised hematological patients[J].Scandinavian Journal of Infectious Diseases,1994,26(2):163-170.
[26]陈之航.要重视肠道致病性大肠埃希菌的检测[J].福州总医院学报,2007(3):148-150.
[27]谢朝云,孙静,熊芸,等.泌尿道与呼吸道感染大肠埃希菌的耐药性分析[J].中华医院感染学杂志,2014,24(16):3933-3935.
[28]李虹敏,冯宪超,徐幸莲,等.肉鸡屠宰加工及冷藏中的微生物污染来源及菌相分析[J].肉类工业,2008(6):33-37.
[29]BAUERMEISTER L J,BOWERS J W,TOWNSEND J C,et al.Validating the efficacy of peracetic acid mixture as an antimicrobial in poultry chillers[J].Journal of Food Protection,2008,71(6):1119-1122.DOI:10.4315/0362-028X-71.6.1119.
[30]SCOTT B R,YANG X,GEORNARAS I,et al.Antimicrobial efficacy of a sulfuric acid and sodium sulfate blend,peroxyacetic acid,and cetylpyridinium chloride against Salmonella on inoculated chicken wings[J].Journal of Food Protection,2015,78(11):1967-1972.DOI:10.4315/0362-028X.JFP-15-170.
[2]华绪川,徐威,朱丽洁.公共卫生安全视角下的家禽交易管理[J].中国家禽,2016,38(17):1-5.
[3]LI Y,ZHUANG S,MUSTAPHA A.Application of a multiplex PCR for the simultaneous detection of Escherichia coli O157:H7,Salmonella and Shigella in raw and ready-to-eat meat products[J].Meat Science,2005,71(2):402-406.DOI:10.1016/j.meatsci.2005.04.013.
[4]MATARAGAS M,SKANDAMIS P,NYCHAS G J E,et al.Modeling and predicting spoilage of cooked,cured meat products by multivariate analysis[J].Meat Science,2007,77(3):348-356.DOI:10.1016/j.meatsci.2007.03.023.
[5]朱恒文,方艳红,王元兰,等.肉鸡屠宰加工生产链中沙门氏菌的污染情况调查及ERIC-PCR溯源[J].食品科学,2012,33(17):48-53.
[6]朱国良,徐景野,章丹阳,等.宁波地区食品中食源性病原菌检测与分析[J].中国公共卫生管理,2014,30(6):907-909.
[7]MASCLAUX F G,SAKWINSKA O,CHARRIERE N,et al.Concentration of airborne Staphylococcus aureus(MRSA and MSSA),total bacteria,and endotoxins in pig farms[J].Annals of Occupational Hygiene,2013,57(5):550-557.DOI:10.1093/annhyq/mes098.
[8]NEHME B,GILBERT Y,LETOURNEAU V,et al.Cultureindependent characterization of archaeal biodiversity in swine confinement building bioaerosols[J].Applied and Environmental Microbiology,2009,75(17):5445-5450.DOI:10.1128/AEM00726-09.
[9]李红梅,白林,姜冬梅,等.基于16S rDNA高通量测序方法检测猪舍空气微生物多样性[J].中国畜牧杂志,2015,51(3):81-84.
[10]SEGATA N,BOERNIGEN D,TICKLE T L,et al.Computational meta’omics for microbial community studies[J].Molecular Systems Biology,2013,9(1):1-15.DOI:10.1038/msb.2013.22.
[11]CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QIIMEallows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.DOI:10.1038/nmeth.f.303.
[12]BOKULICH N A,SUBRAMANIAN S,FAITH J J,et al.Qualityfiltering vastly improves diversity estimates from Illumina amplicon sequencing[J].Nature Methods,2013,10(1):57-59.DOI:10.1038/nmeth.2276.
[13]吕坚成,陈立.畜禽舍空气中灰尘和微生物的危害及防控[J].现代农村科技,2017(1):45.
[14]刘秀萍,唐雨顺,刘永华,等.生猪屠宰加工过程中微生物污染因素及关键控制点分析[J].中国兽医杂志,2014,50(9):72-74.
[15]赵慧,甄少波,任发政,等.生猪屠宰环节微生物状况调查及建议[J]农产品加工(学刊),2012(5):5-6;13.
[16]KIM S A,PARK S H,LEE S I,et al.Assessment of chicken carcass microbiome responses during processing in the presence of commercial antimicrobials using a next generation sequencing approach[J].Scientific Reports,2017,2(23):1-14.DOI:10.1038/srep43354.
[17]KRISTIANSEN A,PEDERSEN K H,NIELSEN P H,et al.Bacterial community structure of a full-scale biofilter treating pig house exhaust air[J].Systematic&Applied Microbiology,2011,34(5):344-352.DOI:10.1016/j.syapm.2010.11.022.
[18]KOTULA K L,PANDYA Y.Bacterial contamination of broiler chickens before scalding[J].Journal of Food Protection,1995,58(12):1326-1329.DOI:10.4315/0362-028X-58.12.1326.
[19]李宗良,赖春颜,梁敏锋,等.葡萄球菌属的临床分布与耐药性分析[J].中华医院感染学杂志,2015,25(12):2684-2686.
[20]ARNOLD H M,SAWYER A M,KOLLEF M H.Use of adjunctive aerosolized antimicrobial therapy in the treatment of Pseudomonas aeruginosa and Acinetobacter baumannii ventilator-associated pneumonia[J].Respiratory Care,2012,57(8):1226-1233.
[21]DENTON M,KERR K G.Microbiological and clinical aspects of infection associated with Stenotrophomonas maltophilia infection[J].Clinical Microbiology Reviews,1998,11(1):57-80.
[22]WAKINO S,IMAI E,YOSHIOKA K,et al.Clinical importance of Stenotrophomonas maltophilia nosocomial pneumonia due to its high mortality in hemodialysis patients[J].Therapeutic Apheresis&Dialysis,2009,13(3):193-198.DOI:10.1111/j.1744-9987.2009.00693.
[23]BROOKE J S.Stenotrophomonas maltophilia:an emerging global opportunistic pathogen[J].Clinical Microbiology Reviews,2012,25(1):2-41.
[24]JANG T N,WANG F D,WANG L S,et al.Xanthomonas maltophilia bacteremia:an analysis of 32 cases[J].Journal of the Formosan Medical Association,1992,91(12):1170-1176.
[25]VICTOR M A,ARPI M,BRUUN B,et al.Xanthomonas maltophilia bacteremia in immunocompromised hematological patients[J].Scandinavian Journal of Infectious Diseases,1994,26(2):163-170.
[26]陈之航.要重视肠道致病性大肠埃希菌的检测[J].福州总医院学报,2007(3):148-150.
[27]谢朝云,孙静,熊芸,等.泌尿道与呼吸道感染大肠埃希菌的耐药性分析[J].中华医院感染学杂志,2014,24(16):3933-3935.
[28]李虹敏,冯宪超,徐幸莲,等.肉鸡屠宰加工及冷藏中的微生物污染来源及菌相分析[J].肉类工业,2008(6):33-37.
[29]BAUERMEISTER L J,BOWERS J W,TOWNSEND J C,et al.Validating the efficacy of peracetic acid mixture as an antimicrobial in poultry chillers[J].Journal of Food Protection,2008,71(6):1119-1122.DOI:10.4315/0362-028X-71.6.1119.
[30]SCOTT B R,YANG X,GEORNARAS I,et al.Antimicrobial efficacy of a sulfuric acid and sodium sulfate blend,peroxyacetic acid,and cetylpyridinium chloride against Salmonella on inoculated chicken wings[J].Journal of Food Protection,2015,78(11):1967-1972.DOI:10.4315/0362-028X.JFP-15-170.