Hass avocados may become contaminated with Salmonella and Listeria monocytogenes at the farm and the packing facility or later during transportation and at retail. In Mexico, avocados are ... HIGHLIGHTS Avocados sold in retail markets in Mexico contain countable levels of microorganisms. Salmonella and L. monocytogenes were isolated from the surface of retail Hass avocados. Salmonella w...

Tomatoes (Solanum lycopersicum) are one of the most commonly consumed fruits worldwide. The fruit can become contaminated with Salmonella and Listeria monocytogenes at various stages of the production and supply chain, and these pathogens may survive under various storage conditions. The effects of relative humidity, temperature, and duration of storage on the attachment and survival of both pathogens on the surface of tomatoes were investigated. Fresh whole Roma tomatoes were inoculated with a cocktail of Salmonella or L. monocytogenes strains and stored at 5, 12, 25, 30, or 35°C for up to 10 days. Every day during storage, relative humidity and temperature were measured and tomatoes were removed to enumerate pathogens cells that were loosely attached (LA; cells were detached from the tomato surface by rinsing) and strongly attached (SA; sonication was required to detach cells from the tomato surface). The attachment strength (SR) was calculated to express the proportion of surviving SA cells on the tomato surface. The initial levels of Salmonella and L. monocytogenes on the tomato surface after inoculation were 6.6 and 6.5 log CFU per tomato for LA cells and 5.1 and 5.6 log CFU per tomato for SA cells, respectively. For both pathogens, the LA levels were higher (P < 0.05) than the SA levels. The LA and SA levels differed significantly as a function of temperature, relative humidity, and duration of storage. The SR for Salmonella was affected by storage time but not temperature, whereas the SR for L. monocytogenes was affected by storage time and temperature and relative humidity (P < 0.05). An understanding of the attachment and survival of Salmonella and L. monocytogenes on tomatoes stored under various temperature conditions may be useful for preventing or reducing the establishment of pathogens and for designing improved decontamination methods.HIGHLIGHTSSurvival of LA cells was highest at 30°C for Salmonella and at 25°C for L. monocytogenes.LA cells of both pathogens declined on the tomato surface at 5°C.Storage at 25 and 30°C and high relative humidity favored the attachment of pathogens.Attachment of L. monocytogenes was high at 12, 30, and 25°C.Attachment of Salmonella was not affected by storage temperature.

Background/Objectives: Listeria monocytogenes is a foodborne pathogen that can infect both humans and animals and cause noninvasive gastrointestinal listeriosis or invasive listeriosis. The objectives of this study were to determine the genetic diversity of L. monocytogenes; the genes associated with its resistance to antibiotics, benzalkonium chloride (BC), and cadmium chloride (CdCl₂); and its biofilm formation. Methods: A total of 132 fresh fruits (44 samples) and vegetables (88 samples) were selected for this study. The genetic diversity of the isolates and the genes associated with their antibiotic resistance were determined using PCR amplification; meanwhile, their levels of susceptibility to antibiotics were determined using the agar diffusion method. Their levels of resistance to BC and CdCl₂ were determined using the minimum inhibitory concentration method, and their capacity for biofilm formation was evaluated using the crystal violet staining method. Results: A total of 17 L. monocytogenes strains were collected: 12.8% (17/132) from fresh fruits and vegetables in this study. The isolates of L. monocytogenes belonged to phylogenetic groups I.1 (29.4% (5/17); serotype 1/2a) and II.2 (70.5% (12/17); serotype 1/2b); strains containing Listeria pathogenicity islands (LIPIs) were also identified at prevalence rates of 100% for LIPI-1 and LIPI-2 (17/17), 29.4% for LIPI-3 (5/17), and 11.7% for LIPI-4 (2/17). The antibiotic susceptibility tests showed that the L. monocytogenes isolates exhibited six different multiresistant patterns, with multiple antibiotic resistance (MAR) index of ≥0.46 (70.5%; 12/17); additionally, the genes Ide, tetM, and msrA, associated with efflux pump Lde, tetracycline, and ciprofloxacin resistance, were detected at 52.9% (9/17), 29.4% (5/17), and 17.6% (3/17), respectively. The phenotypic tests showed that 58.8% (10/17) of cadmium-resistant L. monocytogenes isolates had a co-resistance of 23.5% (4/17) to BC. Finally, all strains of L. monocytogenes exhibited moderate biofilm production. Conclusions: The results of this study contribute to our understanding of the persistence and genetic diversity of L. monocytogenes strains isolated from fresh fruits and vegetables; in addition, their resistance to CdCl₂, which is correlated with co-resistance to BC disinfectant, is helpful for the food industry.

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.