| 31 | 0 | 177 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 探究食品中乳球菌定性及定量检测方法。方法 从市售食品样品中计数并分离乳球菌，结合菌株在平板培养基上和显微镜下形态、生化反应、物质代谢、16S rRNA基因和基质辅助激光解吸电离飞行时间质谱等分析技术，实现对乳球菌分离株的系统鉴定。结果 本研究共采集含乳球菌的食品样品47份。酸奶类样品中乳球菌分离率最高为87.5%(7/8),其次是干酪类为58.3%(14/24)。酸奶和乳酸菌饮料中乳球菌含量均高于1×107 CFU/mL,干酪中有64.3%(9/14)的样品乳球菌含量在1×106～1×107 CFU/g,另35.7%(5/14)处于1×103～1×105 CFU/g范围。饼干中的乳球菌含量高于1×106 CFU/g,黄油和酸奶油在1×104 CFU/g左右。47份样品中共分离出184株菌，其中乳球菌75株，包括59株乳酸乳球菌乳亚种、13株乳酸乳球菌乳亚种(双乙酰型)、3株乳脂乳球菌。结论 建立的乳球菌检测方法可实现食品中乳球菌的计数和鉴定。传统微生物鉴定方法结合分子生物学、质谱等分析技术可实现样品中乳球菌的种、亚种甚至生物型的精确区分。
Abstract：OBJECTIVE To explore qualitative and quantitative detection method for Lactococcus in food samples.METHODS Lactococcus were enumerated and isolated from commercially available food samples,systematic identification of isolates was achieved by combining techniques of colony morphology on plates,microscopic examination,biochemical reactions,metabolic substances,16S rRNA gene sequencing,and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) analysis.RESULTS A total of 47 food samples including yogurt and cheese,that containing Lactococcus were collected in this study.The result indicated that the highest isolation rate of Lactococcus was observed in yogurt samples at 87.5%(7/8),followed by cheese at 58.3%(14/24).The Lactococcus content in both yoghurt and lactic acid beverage was higher than 1×10~7CFU/mL.In cheese,64.3%(9/14) contained Lactococcus between 1×10~6and 1×10~7CFU/g,while the remaining 35.7%(5/14) were within the range of 1×10~3to 1×10~5CFU/g.In biscuit,it was exceeded 1×10~6CFU/g,while butter and sour cream contained approximately 1×10~4CFU/g.184 strains were isolated from the aforementioned samples.59 isolates were identified as L.lactis subsp.lactis,13 were L.lactis subsp lactis biovar diacetylactis and 3 were L.cremoris.CONCLUSION The Lactococcus detection method established in this study enables the enumeration and identification of Lactococcus in food.The combination of traditional microbiological identification method with molecular biology and mass spectrometry techniques enables precise differentiation of Lactococcus species,subspecies,and even biological types in samples.
[1] PASSERINI D,CODDEVILLE M,LE B P,et al.The carbohydrate metabolism signature of Lactococcus lactis strain A12 reveals its sourdough ecosystem origin[J].Appl Environ Microbiol,2013,79(19):5844-5852.
[2] RALLU F,GRUSS A,MAGUIN E.Lactococcus lactis and stress[J].Antonie van Leeuwenhoek,1996,70(2-4):243-251.
[3] ABE K,YUKI M,IMAGAWA Y,et al.Genome-based reclassification of the genus Lactococcus and two novel species Pseudolactococcus yaeyamensis gen.nov.,sp.nov.and Lactovum odontotermitis sp.nov.isolated from the gut of termites[J].Int J Syst Evol Microbiol,2025,75(6):006803.
[4] POUDEL R,THUNELL R K,OBERG C J,et al.Comparison of growth and survival of single strains of Lactococcus lactis and Lactococcus cremoris during Cheddar cheese manufacture[J].J Dairy Sci,2022,105(3):2069-2081.
[5] SCHLEIFER K H,KRAUS J,DVORAK C,et al.Transfer of Streptococcus lactis and related streptococci to the Genus Lactococcus gen.nov[J].Syst Appl Microbiol,1985,6(2):183-195.
[6] TESSIER S,EMENGO I,YODER N,et al.Massive empyema due to Lactococcus garvieae[J].GERMS,2022,12(3):414-418.
[7] KAHALA M,MÄKI M,LEHTOVAARA A,et al.Characterization of starter lactic acid bacteria from the Finnish fermented milk product viili[J].J Appl Microbiol,2008,105(6):1929-1938.
[8] 国家卫生健康委员会，国家市场监督管理总局.食品安全国家标准食品微生物学检验金黄色葡萄球菌检验：GB 4789.10—2016 [S].北京：中国标准出版社，2016.
[9] SMIT G,SMIT B A,ENGELS W J.Flavour formation by lactic acid bacteria and biochemical flavour profiling of cheese products[J].FEMS Microbiol Rev,2005,29(3):591-610.
[10] DRIDER D,BEKAL S,PRÉVOST H.Genetic organization and expression of citrate permease in lactic acid bacteria[J].Genet Mol Res,2004,3(2):273-281.
[11] LOLKEMA J S,POOLMAN B,KONINGS W N.Role of scalar protons in metabolic energy generation in lactic acid bacteria[J].J Bioenerg Biomembr,1995,27(4):467-473.
[12] GARCÍA-QUINTÁNS N,MAGNI C,DE M D,et al.The citrate transport system of Lactococcus lactis subsp.lactis biovar diacetylactis is induced by acid stress[J].Appl Environ Microbiol,1998,64(3):850-857.
[13] SINGHAL N,KUMAR M,KANAUJIA P K,et al.MALDI-TOF mass spectrometry:an emerging technology for microbial identification and diagnosis[J].Front Microbiol,2015,6:791.
[14] NGUYEN D T,VAN H K,CNOCKAERT M,et al.A description of the lactic acid bacteria microbiota associated with the production of traditional fermented vegetables in Vietnam[J].Int J Food Microbiol,2013,163(1):19-27.
[15] ANGELAKIS E,MILLION M,HENRY M,et al.Rapid and accurate bacterial identification in probiotics and yoghurts by MALDI-TOF mass spectrometry[J].J Food Sci,2011,76(8):M568-72.
基本信息:
DOI：10.19813/j.cnki.weishengyanjiu.2026.04.006
中图分类号:R155.5
引用信息:
[1]李鑫,白莉,陈纤蕙,等.食品中乳球菌的计数及精准鉴定技术[J].卫生研究,2026,55(04):567-572+588.DOI:10.19813/j.cnki.weishengyanjiu.2026.04.006.
2026-07-15
2026-07-15