Conditions of In Vitro Biofilm Formation by Serogroups of Listeria monocytogenes Isolated from Hass Avocados Sold at Markets in Mexico

墨西哥市场上销售的哈斯鳄梨中分离的单核细胞增生李斯特菌血清群体外生物膜形成情况

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作者:María Guadalupe Avila-Novoa, Velia Navarrete-Sahagún, Jean Pierre González-Gómez, Carolina Novoa-Valdovinos, Pedro Javier Guerrero-Medina, Ramón García-Frutos, Liliana Martínez-Chávez, Nanci Edid Martínez-Gonzáles, Melesio Gutiérrez-Lomelí

Abstract

Listeria monocytogenes is an important pathogen that has been implicated in foodborne illnesses and the recall of products such as fruit and vegetables. This study determines the prevalence of virulence-associated genes and serogroups and evaluates the effects of different growth media and environmental conditions on biofilm formation by L. monocytogenes. Eighteen L. monocytogenes isolates from Hass avocados sold at markets in Guadalajara, Mexico, were characterized by virulence-associated genes and serogroup detection with PCR. All isolates harbored 88.8% actA, 88.8% plcA, 83.3% mpl, 77.7% inlB, 77.7% hly, 66.6% prfA, 55.5% plcB, and 33.3% inlA. The results showed that 38.8% of isolates harbored virulence genes belonging to Listeria pathogenicity island 1 (LIPI-1). PCR revealed that the most prevalent serogroup was serogroup III (1/2b, 3b, and 7 (n = 18, 66.65%)), followed by serogroup IV (4b, 4d-4e (n = 5, 27.7%)) and serogroup I (1/2a-3a (n = 1, 5.5%)). The assessment of the ability to develop biofilms using a crystal violet staining method revealed that L. monocytogenes responded to supplement medium TSBA, 1/10 diluted TSBA, and TSB in comparison with 1/10 diluted TSB (p < 0.05) on polystyrene at 240 h (p < 0.05). In particular, the biofilm formation by L. monocytogenes (7.78 ± 0.03-8.82 ± 0.03 log10 CFU/cm2) was significantly different in terms of TSBA on polypropylene type B (PP) (p < 0.05). In addition, visualization by epifluorescence microscopy, scanning electron microscopy (SEM), and treatment (DNase I and proteinase K) revealed the metabolically active cells and extracellular polymeric substances of biofilms on PP. L. monocytogenes has the ability to develop biofilms that harbor virulence-associated genes, which represent a serious threat to human health and food safety.

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