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从食品接触表面分离出的生物膜的遗传和成分分析。

Genetic and compositional analysis of biofilm formed by isolated from food contact surfaces.

作者信息

Avila-Novoa María Guadalupe, Solis-Velazquez Oscar Alberto, Guerrero-Medina Pedro Javier, González-Gómez Jean-Pierre, González-Torres Berenice, Velázquez-Suárez Noemí Yolanda, Martínez-Chávez Liliana, Martínez-Gonzáles Nanci Edid, De la Cruz-Color Lucia, Ibarra-Velázquez Luz María, Cardona-López Marco Antonio, Robles-García Miguel Ángel, Gutiérrez-Lomelí Melesio

机构信息

Centro de Investigación en Biotecnología Microbiana y Alimentaria, Departamento de Ciencias Básicas, Centro Universitario de la Ciénega, Universidad de Guadalajara, Ocotlán, Jalisco, Mexico.

Laboratorio Nacional para la Investigación en Inocuidad Alimentaria (LANIIA), Centro de Investigación en Alimentación y Desarrollo, A.C. (CIAD), Culiacán, Sinaloa, Mexico.

出版信息

Front Microbiol. 2022 Dec 2;13:1001700. doi: 10.3389/fmicb.2022.1001700. eCollection 2022.

Abstract

INTRODUCTION

is an important pathogen that can form biofilms on food contact surfaces (FCS) in the dairy industry, posing a serious food safety, and quality concern. Biofilm is a complex system, influenced by nutritional-related factors that regulate the synthesis of the components of the biofilm matrix. This study determines the prevalence of biofilm-associated genes and evaluates the development under different growth conditions and compositions of biofilms produced by .

METHODS

Biofilms were developed in TSB, TSBG, TSBNaCl, and TSBGNaCl on stainless-steel (SS), with enumeration at 24 and 192 h visualized by epifluorescence and scanning electron microscopy (SEM). The composition of biofilms was determined using enzymatic and chemical treatments and confocal laser scanning microscopy (CLSM).

RESULTS AND DISCUSSION

A total of 84 (SA1-SA84) strains were collected from 293 dairy industry FCS (FCS-stainless steel [ = 183] and FCS-polypropylene [ = 110]) for this study. The isolates harbored the genes (66%), (53%), (52%), (38%), (20%), and (9.5%). 99. In particular, the biofilm formed by -positive onto SS showed a high cell density in all culture media at 192 h in comparison with the biofilms formed at 24 h ( < 0.05). Epifluorescence microscopy and SEM revealed the metabolically active cells and the different stages of biofilm formation. CLSM analysis detected extracellular polymeric of biofilms on SS, such as eDNA, proteins, and polysaccharides. Finally, the level of detachment on being treated with DNase I (44.7%) and NaIO 4(42.4%) was greater in the biofilms developed in TSB compared to culture medium supplemented with NaCl at 24 h; however, there was no significant difference when the culture medium was supplemented with glucose. In addition, after treatment with proteinase K, there was a lower level of biomass detachment (17.7%) of the biofilm developed in TSBNaCl ( < 0.05 at 24 h) compared to that in TSB, TSBG, and TSBGNaCl (33.6, 36.9, and 37.8%, respectively). These results represent a deep insight into the composition of biofilms present in the dairy industry, which promotes the development of more efficient composition-specific disinfection strategies.

摘要

引言

是一种重要的病原体,可在乳制品行业的食品接触表面(FCS)上形成生物膜,引发严重的食品安全和质量问题。生物膜是一个复杂的系统,受营养相关因素影响,这些因素调节生物膜基质成分的合成。本研究确定了生物膜相关基因的流行情况,并评估了在不同生长条件下由产生的生物膜的发育情况以及生物膜的组成。

方法

在不锈钢(SS)上于胰酪大豆胨肉汤(TSB)、胰酪大豆胨葡萄糖肉汤(TSBG)、胰酪大豆胨氯化钠肉汤(TSBNaCl)和胰酪大豆胨葡萄糖氯化钠肉汤(TSBGNaCl)中培养生物膜,在24小时和192小时进行计数,通过落射荧光显微镜和扫描电子显微镜(SEM)观察。使用酶处理、化学处理和共聚焦激光扫描显微镜(CLSM)确定生物膜的组成。

结果与讨论

本研究从293个乳制品行业的食品接触表面(FCS - 不锈钢[ = 183]和FCS - 聚丙烯[ = 110])收集了总共84株(SA1 - SA84)菌株。分离株携带基因(66%)、(53%)、(52%)、(38%)、(20%)和(9.5%)。99. 特别是,与24小时形成的生物膜相比,在192小时时,阳性在SS上形成的生物膜在所有培养基中显示出高细胞密度(<0.05)。落射荧光显微镜和SEM揭示了代谢活跃的细胞和生物膜形成的不同阶段。CLSM分析检测到SS上生物膜的细胞外聚合物,如胞外DNA、蛋白质和多糖。最后,在24小时时,与添加氯化钠的培养基相比,用脱氧核糖核酸酶I(44.7%)和高碘酸钠(42.4%)处理后,TSB中形成的生物膜的脱落水平更高;然而,当培养基添加葡萄糖时没有显著差异。此外与TSB、TSBG和TSBGNaCl(分别为33.6%、36.9%和37.8%)相比,用蛋白酶K处理后,TSBNaCl中形成的生物膜的生物量脱落水平较低(24小时时<0.05)(分别为17.7%)。这些结果深入了解了乳制品行业中生物膜的组成,有助于开发更有效的针对特定组成的消毒策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8fa/9755592/90b096f42c7d/fmicb-13-1001700-g001.jpg

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