Wet steam method for reducing attached cells of Salmonella and Listeria monocytogenes on Hass avocado.

The consumption of avocados and their products has been linked to outbreaks of illness caused by Salmonella enterica and Listeria monocytogenes. These pathogens have been isolated from avocados collected from farms and markets. After contact with the avocado epicarp, the cells of Salmonella and L. monocytogenes can become loosely attached (LA) by suspension in a film of water and attraction by electrostatic forces, or strongly attached (SA) by physical and irreversible attachment mechanisms. Attached cells may have greater resistance to agents used to decontaminate the fruit. The effect of applying wet steam (WS) to the epicarp of Hass avocados on the reduction LA and SA counts of Salmonella and L. monocytogenes was evaluated as a function of the exposure time. The inoculated avocados were washed and exposed to WS for 30, 45, and 60 s inside a treatment chamber. Salmonella was found to be more susceptible to WS than L. monocytogenes. The efficacy of steam in reducing LA and SA cell numbers was similar for both pathogens. Steaming avocados for 60 s reduced LA Salmonella and L. monocytogenes cells by 4.6 and 4.8 log CFU/avocado, whereas SA cells were decreased by 5.2 and 4.4 log CFU/avocado, respectively.•Steaming the avocados for 60 s produced the greatest reduction in loosely and strongly attached cells for both pathogens.•Wet steam treatment efficiently eliminated the loosely and strongly attached cells of both pathogens.•The Listeria monocytogenes attached cells showed greater resistance to steam treatment than Salmonella.

Genomic Insights into Listeria monocytogenes: Organic Acid Interventions for Biofilm Prevention and Control.

Listeria monocytogenes is an important pathogen that has been implicated in foodborne illness. The aim of the present study was to investigate the diversity of virulence factors associated with the mechanisms of pathogenicity, persistence, and formation of biofilm L. monocytogenes by tandem analysis of whole-genome sequencing. The lineages that presented L. monocytogenes (LmAV-2, LmAV-3, and LmAV-6) from Hass avocados were lineages I and II. Listeria pathogenicity island 1 (LIPI-1) and LIPI-2 were found in the isolates, while LIPI-3 and Listeria genomic island (LGI-2) only was in IIb. Stress survival island (SSI-1) was identified in lineage I and II. In the in silico analysis, resistance genes belonging to several groups of antibiotics were detected, but the bcrABC and transposon Tn6188 related to resistance to quaternary ammonium salts (QACs) were not detected in L. monocytogenes. Subsequently, the anti-L. monocytogenes planktonic cell effect showed for QACs (MIC = 6.25 ppm/MBC = 100 ppm), lactic acid (MBC = 1 mg/mL), citric acid (MBC = 0.5 mg/mL) and gallic acid (MBC = 2 mg/mL). The anti-biofilm effect with organic acids (22 °C) caused a reduction of 4-5 log10 cfu/cm2 after 10 min against control biofilm L. monocytogenes formed on PP than SS. This study is an important contribution to understanding the genomic diversity and epidemiology of L. monocytogenes to establish a control measure to reduce the impact on the environment and the consumer.

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

Introduction: Staphylococcus aureus 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 S. aureus. 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 S. aureus (SA1-SA84) strains were collected from 293 dairy industry FCS (FCS-stainless steel [n = 183] and FCS-polypropylene [n = 110]) for this study. The isolates harbored the genes sigB (66%), sar (53%), agrD (52%), clfB/clfA (38%), fnbA/fnbB (20%), and bap (9.5%). 99. In particular, the biofilm formed by bap-positive S. aureus onto SS showed a high cell density in all culture media at 192 h in comparison with the biofilms formed at 24 h (p < 0.05). Epifluorescence microscopy and SEM revealed the metabolically active cells and the different stages of biofilm formation. CLSM analysis detected extracellular polymeric of S. aureus 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 (p < 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 S. aureus biofilms present in the dairy industry, which promotes the development of more efficient composition-specific disinfection strategies.

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

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.

Salmonella, Listeria monocytogenes, and Indicator Microorganisms on Hass Avocados Sold at Retail Markets in Guadalajara, Mexico.

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 frequently sold in bulk at retail markets, where they are stored at room temperature for several hours or days and exposed to potential sources of microorganisms. These conditions may favor the entry, adhesion, survival, and biofilm formation of Salmonella and L. monocytogenes. The aim of this study was to determine the occurrence of Salmonella, L. monocytogenes, and other Listeria species and the levels of indicator microorganisms on the surface of avocados sold at retail markets. A total of 450 samples (Persea americana var. Hass) were acquired from retail markets located in Guadalajara, Mexico. One group of 225 samples was evaluated for the presence of Salmonella and for enumeration of aerobic plate counts, yeasts and molds, Enterobacteriaceae, coliforms, and Escherichia coli. The other 225 samples were processed for isolation of L. monocytogenes and other Listeria species. Microbial counts (log CFU per avocado) were 4.3 to 9.0 for aerobic plate counts, 3.3 to 7.1 for yeasts and molds, 3.3 to 8.2 for Enterobacteriaceae, 3.3 to 8.4 for coliforms, and 3.3 to 6.2 for E. coli. Eight samples (3.5%) were positive for Salmonella. Listeria spp. and L. monocytogenes were detected in 31 (13.8%) and 18 (8.0%) of 225 samples, respectively. Listeria innocua, Listeria welshimeri, and Listeria grayi were isolated from 7.6, 1.3, and 0.9% of samples. These results indicate that avocados may carry countable levels of microorganisms and could be a vehicle for transmission of Salmonella and L. monocytogenes.