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微生物学报

植物乳杆菌培养上清对不同血清型沙门氏菌的抑制效果及作用机理
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国家自然科学基金(32002212);浙江省自然科学基金(LQ21C170001);浙江农林大学科研发展项目(2020FR003);浙江省领军型创新创业团队项目(2020R01015);浙江农林大学学生科研训练项目(2021KX0114)


Inhibitory mechanism of Lactobacillus plantarum culture supernatant on Salmonella of different serotypes
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    摘要:

    【目的】探究植物乳杆菌培养上清(Lactobacillus plantarum culture supernatant,LPC)对3种血清型沙门氏菌猪霍乱(Salmonellacholerae,SC)、肠炎(Salmonella enteritidis,SE)和鸡白痢(Salmonella pullorum,SP)的生长和致病性的抑制作用效果及机理。【方法】将2%LPC与3种沙门氏菌分别共培养后,采用比浊法及牛津杯抑菌圈试验检测沙门氏菌生长情况及LPC中的主要抑菌物质,使用实时荧光定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)探究沙门氏菌致病性相关基因表达水平,最后通过结晶紫染色法检测沙门氏菌的生物被膜。【结果】2%LPC能够显著抑制3种沙门氏菌的生长,其作用效果与庆大霉素(gentamicin,GM)相近且对SE的生长抑制效果优于GM,其主要抑菌物质为有机酸;2%LPC对3株沙门氏菌SPI-1编码的主要毒力基因(InvAInvFSopESopBSipBHilASipA)、SPI-2毒力基因(SopD2)、菌毛相关基因(FliFLpfASefAFimF)和鞭毛相关基因(FlhDFliCFliD)表达均有显著抑制效果且抑制效果与GM相近;2%LPC对3株沙门氏菌的生物被膜形成均有显著抑制效果,抑菌率12h为40%-50%、24h为60%-80%,且作用效果与GM相似;对比发现,2%LPC对SE的生长及致病性的抑制效果优于SC与SP。【结论】2%LPC对3种血清型沙门氏菌的生长性能及致病性均有显著抑制效果,且对肠炎沙门氏菌抑制效果最佳。

    Abstract:

    [Objective] Our study investigated the inhibitory effect and pathogenicity of Lactobacillus plantarum culture supernatant (LPC) on Salmonella cholerae (SC), Salmonella enteritidis (SC) and Salmonella pullorum (SP). [Methods] Three serotypes of Salmonella enterica were co-cultured with 2% LPC. Salmonella growth and antibacterial compounds were identified using turbidimetry and the Oxford cup antimicrobial zone. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of pathogenicity-related genes in Salmonella, and crystal violet staining was used to identify the biofilm of Salmonella. [Results] 2% LPC reduced the development of three Salmonella strains considerably, with an efficacy comparable to gentamicin (GM) and even better than SE. Additionally, the main antibacterial substance was organic acids. 2% LPC markedly suppressed the expressions of the main virulence genes encoded by SPI-1 (InvA, InvF, SopE, SopB, SipB, HilA and SipA), SPI-2 (SopD2), pili related genes (FliF, LpfA, SefA and FimF) and flagellum related genes (FlhD, FlIC and FliD) and the inhibitory effect was similar to GM. 2% LPC had significant inhibitory effect on biofilm formation of the three Salmonella strains, the inhibition rate was 40%–50% at 12 h and 60%–80% at 24 h. Compared with SC and SP, 2% LPC was superior to inhibit SE. [Conclusion] 2% LPC has significant inhibitory effects on the growth and pathogenicity of Salmonella serotypes, particlarly SE.

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李东昱,胡爱心,黄雯霞,舒鑫,刘金松,张瑞强,许英蕾,杨彩梅,吴艳萍. 植物乳杆菌培养上清对不同血清型沙门氏菌的抑制效果及作用机理. 微生物学报, 2023, 63(1): 220-232

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  • 收稿日期:2022-04-16
  • 最后修改日期:2022-05-20
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  • 在线发布日期: 2023-01-13
  • 出版日期: 2023-01-04