Genomic diversity of Salmonella enterica isolated from raw chicken at retail establishments in Mexico.

The genomic diversity of circulating non-typhoidal Salmonella in raw chicken was investigated in three states of central Mexico. A total of 192 S. enterica strains from chicken meat samples collected at supermarkets, fresh markets, and butcher shops were analyzed by whole-genome sequencing. The serovar distribution, occurrence of genes encoding for antimicrobial resistance, metal resistance, biocide resistance, plasmids and virulence factors, and clonal relatedness based on single nucleotide polymorphism (SNP) analysis were investigated. Serovars Infantis, Schwarzengrund and Enteritidis predominated among twenty identified. The distribution of serovars and proportion of AMR genes was different according to the state, year, season, and retail establishment (p < 0.001). Genes encoding metals resistance were identified in all the strains. A total of 145 virulence genes were identified and strains were classified into 32 virulotypes; serovars Infantis, Typhimurium, and Enteritidis showed the highest number of virulence genes. The strains matched 34 SNP clusters in the NCBI Pathogen Detection server and 59 %, which corresponded to Infantis, Schwarzengrund, Saintpaul, and Enteritidis, were associated with five major clusters and matched with chicken, environmental and clinical isolates from at least three countries. These results provide useful information to understand the epidemiology of Salmonella, conduct microbial risk assessment, and design risk-based control measures.

Genomic analysis of the MLST population structure and antimicrobial resistance genes associated with Salmonella enterica in Mexico.

Salmonella enterica is one of the most commonly reported foodborne pathogens by public health agencies worldwide. In this study, the multilocus sequence typing (MLST) population structure and frequency of antimicrobial resistance (AMR) genes were evaluated in S. enterica strains from Mexico (n = 2561). The most common sources of isolation were food (44.28%), environment (27.41%), animal-related (24.83%), and human (3.48%). The most prevalent serovars were Newport (8.51%), Oranienburg (7.03%), Anatum (5.78%), Typhimurium (5.12%), and Infantis (4.57%). As determined by the 7-gene MLST scheme, the most frequent sequence types were ST23, ST64, and ST32. The core genome MLST scheme identified 132 HC2000 and 195 HC900 hierarchical clusters, with the HC2000_2 cluster being the most prevalent in Mexico (n = 256). A total of 78 different AMR genes belonging to 13 antimicrobial classes were detected in 638 genomic assemblies of S. enterica. The most frequent class was aminoglycosides (31.76%), followed by tetracyclines (12.53%) and sulfonamides (11.91%). These results can help public health agencies in Mexico prioritize their efforts and resources to increase the genomic sequencing of circulating Salmonella strains. Additionally, they provide valuable information for local and global public health efforts to reduce the impact of foodborne diseases and AMR.

Risk ranking of food categories associated with Salmonella enterica contamination in the central region of Mexico.

To prevent and control foodborne diseases, there is a fundamental need to identify the foods that are most likely to cause illness. The goal of this study was to rank 25 commonly consumed food products associated with Salmonella enterica contamination in the Central Region of Mexico. A multicriteria decision analysis (MCDA) framework was developed to obtain an S. enterica risk score for each food product based on four criteria: probability of exposure to S. enterica through domestic food consumption (Se); S. enterica growth potential during home storage (Sg); per capita consumption (Pcc); and food attribution of S. enterica outbreak (So). Risk scores were calculated by the equation Se*W1 +Sg*W2 +Pcc*W3 +So*W4 , where each criterion was assigned a normalized value (1-5) and the relative weights (W) were defined by 22 experts' opinion. Se had the largest effect on the risk score being the criterion with the highest weight (35%; IC95% 20%-60%), followed by So (24%; 5%-50%), Sg (23%; 10%-40%), and Pcc (18%; 10%-35%). The results identified chicken (4.4 ± 0.6), pork (4.2 ± 0.6), and beef (4.2 ± 0.5) as the highest risk foods, followed by seed fruits (3.6 ± 0.5), tropical fruits (3.4 ± 0.4), and dried fruits and nuts (3.4 ± 0.5), while the food products with the lowest risk were yogurt (2.1 ± 0.3), chorizo (2.1 ± 0.4), and cream (2.0 ± 0.3). Approaches with expert-based weighting and equal weighting showed good correlation (R2  = 0.96) and did not show significant differences among the ranking order in the top 20 tier. This study can help risk managers select interventions and develop targeted surveillance programs against S. enterica in high-risk food products.

Genotypic and phenotypic quantitative microbial risk assessment model of human salmonellosis related to the consumption of chicken meat in the central region of Mexico.

Chicken meat is often associated withSalmonella entericacontamination worldwide. This study proposes a risk assessment model for human salmonellosis linked to the domestic consumption of chicken meat in the central region of Mexico, incorporating genotypic and phenotypic data. SixS. entericagroups were used, considering the presence of specific virulence genes and multidrug resistance (MDR). Sixteen exposure scenarios were established considering retail point (RP1 = fresh market/butcher shop; RP2 = mini-super/supermarket), transportation, home storage, cooking, and cross-contamination. The model predicted a mean annual salmonellosis cases of 66,754 due to chicken consumption (CI95% 10775-231606). The mean probability of illness (Pill) among the exposure scenarios ranged from 2.5 × 10-9 to 3.7 × 10-6, 7.7 × 10-8 to 1.1 × 10-4, and 6.7 × 10-4 to 7.8 × 10-2 for low, moderate, and high virulence groups. Exposure scenarios with the highest Pill were not responsible for most cases due to their low frequency of occurrence. The high virulence/ MDR group was responsible for most cases (66.5 %), despite the low S. enterica prevalence (RP1 0.5 % and RP2 5.0 %). The years lost due to disability (YLD) value for MDR was 2.6 × higher than for non-MDR. Spearman rank showed that the inputs with higher influence on the variability of salmonellosis depended on the type of exposure scenario. For example, the cooking temperature and time had the most significant influence in the scenarios where S. enterica can survive after cooking. Including the microbial genotypic and phenotypic characteristics in risk assessment modeling highlights the importance of focusing on high-virulent and MDR strains, which are not the most frequent but represent the highest public health risk.

Comparative Study between Two Simple Synthesis Methods for Obtaining Green Gold Nanoparticles Decorating Silica Particles with Antibacterial Activity.

The SiO2 particles system is one of the most common ways to protect colloidal metal systems, such as gold nanoparticles, from aggregation and activity loss due to their high chemical stability and low reactivity. In this study, silica green gold nanoparticles (AuNPs synthesized with mullein extract) were fabricated using two different sol-gel methods. The nanoparticles were characterized by Scanning Electron Microscopy (SEM), X-ray diffraction (XRD), Fourier Transformed Infrared (FTIR), and the antibacterial activity against pathogens (Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, and Salmonella enterica). Synthesis-1 nanoparticles had a kidney-shaped form and uniform distribution, while synthesis-2 nanoparticles had a spherical and non-uniform form. Characterization showed that temperature is an important factor in the distribution of AuNPs in silica; a decrease allowed the formation of Janus-type, and an increase showed a higher concentration of gold in energy-dispersive spectroscopy (EDS) analysis. Overall, similar bands of the two synthesis silica nanoparticles were observed in FTIR, while XRD spectra showed differences in the preferential growth in AuNPs depending on the synthesis. Higher antibacterial activity was observed against S. aureus, which was followed by L. monocytogenes. No differences were observed in the antibacterial activity between the two different sol-gel methods.

Detection, quantification, and characterization of Salmonella enterica in mango, tomato, and raw chicken purchased in the central region of Mexico.

To estimate human exposure to Salmonella enterica, it is essential to understand the pathogen distribution and characteristics. Prevalence and concentration of S. enterica were determined in mango, tomato, and raw chicken samples purchased in three states (Aguascalientes, Querétaro, and Guadalajara) located in the central region of Mexico during two seasons. In addition, S. enterica isolates were characterized by absence/presence of 13 virulence genes (chromosomal, prophage, and plasmid) and resistance to 14 antibiotics. A total of 300 samples of mango, 272 of tomato, and 354 of raw chicken were analyzed. The mean of the prevalence (24.9%) and concentration (-0.61 Log MPN/g) of S. enterica in chicken was higher than in mango (1.3%, -1.7 Log MPN/g) and tomato (1.1%, -1.7 Log MPN). Among S. enterica isolates (284), there were 7 different virulotypes, belonging 68.7% of isolates to V2; there was high variability in the presence of mobile genetic elements. The occurrence of specific mobile elements ranged from 81.4% to 11.3% among isolates. Among the isolates, 91.5% were resistant to at least one antibiotic with ampicillin being the most frequent; 54.9% of isolates were multidrug resistant. Data from this study can be used for quantitative microbial risk assessment of S. enterica related to mango, tomato, and raw chicken consumption in the central region of Mexico. PRACTICAL APPLICATION: Data on the prevalence and concentration of Salmonella enterica obtained in this study can be used to estimate the exposure assessment for the consumption of mango, tomato, and chicken in the central region of Mexico. In addition, the characteristics of the S. enterica isolates could be used to select representative strains for future studies to evaluate the intraspecies variability.

Salmonella enterica in Mexico 2000-2017: Epidemiology, Antimicrobial Resistance, and Prevalence in Food.

In Mexico, information of Salmonella enterica cases linked to food consumption is scarce. The objective of this article was to assess how S. enterica affect public health in Mexico. To conduct this study, data on the epidemiology of nontyphoidal S. enterica (NTS), Salmonella Typhi, and Salmonella Paratyphi A collected from 2000 to 2017 through the National Epidemiological Surveillance System of Mexico (Sistema Nacional de Vigilancia Epidemiológica de Mexico [SINAVE]) were used. Geographical distribution, season, age groups, and gender were variables considered to analyze S. enterica incidence. An estimation of cases caused by S. enterica in Mexico was calculated while considering data underestimation and the proportion of foodborne diseases. Information of the prevalence of the pathogen in food and the antimicrobial resistance of isolates from food and human cases were obtained from published studies. Outbreaks of S. enterica derived from imported Mexican products in the Unites States are discussed. In 2017, the numbers of reported cases of NTS (92,013) were two and seven times higher than the reported cases of Salmonella Typhi (45,280) and Salmonella Paratyphi A (12, 458). The NTS incidence was higher in lower socioeconomic Mexican regions. The gaps in the surveillance system make it impossible to establish a reliable tendency among age groups, geographical distribution, and gender. In 2017, the estimated frequency of NTS foodborne cases was 49 times higher than that reported in SINAVE, whereas for Salmonella Typhi and Salmonella Paratyphi A it was 23 times. Fresh meat showed the highest prevalence of S. enterica, and most of their isolates had multidrug resistance. Salmonella Typhimurium was the most common serotype isolated from human cases and food. Food safety agencies in Mexico need to prioritize efforts and resources to establish guidelines to ensure the absence of S. enterica in food.

Food Consumer Behavior and Salmonella Exposure Self-Perception in the Central Region of Mexico.

In Mexico, there is limited information regarding food consumer behaviors at home. The knowledge of food consumption and food handling practices can help to determine more realistically the exposure to different hazards, such as Salmonella spp. on the population. The main goal of this study was to characterize the food consumption habits of five food groups (fruits and vegetables, fresh meat, dairy products, processed meat products, and seafood), food handling practices, and Salmonella exposure self-perception of the population of the central region of Mexico using an online survey. A total of 1,199 surveys were analyzed. The group of fruits and vegetables is the most consumed and one of the most related to the consumer self-perception of suffering from salmonellosis. Mexican consumers regularly use refrigeration as the principal food storage method for the five food groups (42.2% to 90.8%), and generally, they store the products between 2 and 3 days (26.5% to 38.1%). A total of 86.4% of consumers reported that they always wash their hands prior to food preparation. A total of 16.9% and 13.0% use the same cutting board and knife, respectively, to cut more than one product without applying sanitization. Men, people between 20 and 24 years, and people between 60 and 64 years, had the highest risk food handling practices at home. Finally, age (P < 0.0001), education level (P = 0.0004), and suffering from gastrointestinal diseases (P = 0.0005) showed significant effects with self-perception of having salmonellosis. PRACTICAL APPLICATION: Data presented in this study could be used in future risk and exposure assessment research to evaluate the public health risk from the consumption of contaminated food. In addition, it could be used by government agencies to design education campaigns in food safety area.

Antibacterial Activities of Hibiscus sabdariffa Extracts and Chemical Sanitizers Directly on Green Leaves Contaminated with Foodborne Pathogens.

Leafy greens have been associated with foodborne disease outbreaks in different countries. To decrease microbial contamination of leafy greens, chemical agents are commonly used; however, a number of studies have shown these agents to have limited antimicrobial effect against pathogenic bacteria on vegetables. The objective of this study was to compare the antibacterial effect of Hibiscus sabdariffa calyx extracts (water, methanol, acetone, and ethyl acetate), sodium hypochlorite, acetic acid, and colloidal silver against foodborne bacteria on leafy greens. Thirteen foodborne bacteria were used in the study: Listeria monocytogenes, Shigella flexneri, Salmonella serotypes Typhimurium Typhi, and Montevideo, Staphylococcus aureus, Escherichia coli O157:H7, five E. coli pathotypes (Shiga toxin-producing, enteropathogenic, enterotoxigenic, enteroinvasive, and enteroaggregative), and Vibrio cholerae O1. Each foodborne bacterium was separately inoculated on romaine lettuce, spinach, and coriander leaves. Separately, contaminated leafy greens were immersed in four hibiscus extracts and in sanitizers for 5 min. Next, green leaves were washed with sterile tap water. Separately, each green leaf was placed in a bag that contained 0.1% sterile peptone water and was rubbed for 2 min. Counts were done by plate count using appropriate dilutions (in sterile peptone water) of the bacterial suspensions spread on Trypticase soy agar plates and incubated at 35 ± 2°C for 48 h. Statistically significant differences ( P < 0.05) were calculated with an analysis of variance and Duncan's test. All 13 foodborne bacteria attached to leafy greens. Roselle calyx extracts caused a significantly greater reduction ( P < 0.05) in concentration of all foodborne bacteria on contaminated romaine lettuce, spinach, and coriander than did the sodium hypochlorite, colloidal silver, and acetic acid. Dry roselle calyx extracts may potentially be a useful addition to disinfection procedures for romaine lettuce, spinach, and coriander.

An Improve Protocol for PCR Using LM1 and LM2 Primers for Listeria monocytogenes Detection in Food Matrices.

Several studies have observed that a conventional PCR protocol using primers LM1 and LM2 for the identification of gene hlyA Listeria monocytogenes generates non-specific PCR amplifications and false positives. For this reason in this study, we provide a modified PCR protocol that improves the specificity of the LM1 and LM2 primers.

Attachment of 13 Types of Foodborne Bacteria to Jalapeño and Serrano Peppers and Antibacterial Effect of Roselle Calyx Extracts, Sodium Hypochlorite, Colloidal Silver, and Acetic Acid against These Foodborne Bacteria on Peppers.

Chili peppers are a very important crop in Mexico. However, these peppers have been associated with Salmonella infection outbreaks in the United States, and Salmonella and diarrheagenic Escherichia coli pathotypes have been isolated from jalapeño and serrano peppers in Mexico. To decrease microbial contamination of fruits and vegetables, chemical agents are commonly used; however, chemical agents used to eliminate pathogenic bacteria on vegetables have a limited antimicrobial effect. Roselle ( Hibiscus sabdariffa ) calyces have been reported to have an antimicrobial effect on pathogenic bacteria. In the present study, the antibacterial effect of four roselle calyx extracts (water, methanol, acetone, and ethyl acetate), sodium hypochlorite, colloidal silver, and acetic acid against foodborne bacteria was evaluated on contaminated jalapeño and serrano peppers. The 13 types of foodborne bacteria evaluated were Listeria monocytogenes , Shigella flexneri , Salmonella Typhimurium, Salmonella Typhi, Salmonella Montevideo, Staphylococcus aureus , E. coli O157:H7, five E. coli pathotypes (Shiga toxin producing, enteropathogenic, enterotoxigenic, enteroinvasive, and enteroaggregative), and Vibrio cholerae O1. All 13 types attached to both pepper types, with no significant differences in attachment between jalapeño and serrano peppers. Roselle calyx extract treatment resulted in a greater reduction in levels of all foodborne bacteria than did treatment with sodium hypochlorite, colloidal silver, and acetic acid on both pepper types. Roselle calyx extracts may be a useful for disinfection of chili peppers in the field, processing plants, restaurants, and homes.