ABSTRACT
Developing countries such as Ecuador carry a heavy food safety burden but reports on the microbiological quality of their foods are scarce. In this investigation, the microbial diversity of 10 high-risk and mass-consumption street-vended foods including bolones, encebollado, food dressings, ceviche, chopped fruits, fruit juices, fruit salads, cheese, raw chicken, and ground beef in Quito, Guayaquil, and Cuenca, three major population centers in Ecuador, were evaluated using 16S rRNA gene High Throughput Sequencing. In total, 1,840 amplicon sequence variants (ASVs) were classified into 23 phyla, 253 families, 645 genera, and 829 species. In the tested food samples, Proteobacteria and Firmicutes were the most abundant phyla accounting for 97.41% of relative abundance (RA). At genus level, 10 dominant genera were identified: Acinetobacter (12.61% RA), Lactococcus (12.08% RA), Vibrio (8.23% RA), Weissella (7.43% RA), Aeromonas (6.18% RA), Photobacterium (6.32% RA), Pseudomonas (3.92% RA), Leuconostoc (3.51% RA), Klebsiella (3.49% RA), and Cupriavidus (2.86% RA). The highest microbial diversity indices were found in raw chicken, encebollados, fruit salads, and fruit juices from Guayaquil and Cuenca. From sampled foods, 29 species were classified as food spoilage bacteria and 24 as opportunistic pathogenic bacteria. Two groups associated with human diseases were identified, including 11 enteric species and 26 species of fecal bacteria. The occurrence of recognized and opportunistic pathogenic bacteria, as well as enteric and fecal microorganisms, in the street-vended foods indicated extensive risks for the consumers' health. This study demonstrated the application of culture-independent amplicon sequencing in providing a more comprehensive view of microbial safety for street-vended food, which could be a useful tool to facilitate the control of foodborne diseases.
Subject(s)
Food Microbiology , Vibrio , Animals , Cattle , Humans , RNA, Ribosomal, 16S/genetics , Ecuador , Food Safety , Vibrio/geneticsABSTRACT
Imported papayas from Mexico have been implicated in multiple salmonellosis outbreaks in the United States in recent years. While postharvest washing is a critical process to remove latex, dirt, and microbes, it also has the potential of causing cross-contamination by foodborne pathogens, with sponge or other fibrous rubbing tools often questioned as potential harboring or transmitting risk. In this study, Salmonella inactivation and cross-contamination via sponges and microfiber wash mitts during simulated papaya washing and cleaning were investigated. Seven washing treatments (wash without sanitizer; wash at free chlorine 25, 50, and 100 mg/L, and at peracetic acid 20, 40, and 80 mg/L), along with unwashed control, were evaluated, using Salmonella strains with unique antibiotic markers differentially inoculated on papaya rind (serovars Typhimurium, Heidelberg, and Derby) and on wash sponge or microfiber (serovars Typhimurium, Newport, and Braenderup). Salmonella survival and transfer on papaya and on sponge/microfiber, and in wash water were detected using selective plating or enrichment. The washing and cleaning process reduced Salmonella on inoculated papayas by 1.69-2.66 and 0.69-1.74 log for sponge and microfiber cleaning, respectively, with the reduction poorly correlated to sanitizer concentration. Salmonella on inoculated sponge or microfiber was under detection limit (1.00 log CFU/cm2) by plate count, but remained recoverable by selective enrichment. Transference of Salmonella from inoculated papaya to sponge/microfiber, and vice versa, could be detected sporadically by selective enrichment. Sponge/microfiber mediated Salmonella cross-contamination from inoculated to uninoculated papayas was frequently detectable by selective enrichment, but rendered undetectable by wetting sponge/microfiber in sanitizing wash water (FC 25-100 mg/L or PAA 20-80 mg/L) between washing different papaya fruits. Therefore, maintaining adequate sanitizer levels and frequently wetting sponge/microfiber in sanitizing wash water can effectively mitigate risks of Salmonella cross-contamination associated with postharvest washing, especially with regard to the use of sponge or microfiber wash mitts.
Subject(s)
Carica/microbiology , Chlorine/pharmacology , Disinfectants/pharmacology , Food Handling/instrumentation , Peracetic Acid/pharmacology , Porifera/microbiology , Salmonella typhimurium/drug effects , Animals , Food Contamination/analysis , Food Contamination/prevention & control , Food Handling/methods , Fruit/microbiology , Mexico , Salmonella typhimurium/growth & developmentABSTRACT
ABSTRACT: Bacterial foodborne diseases are among the most important public health issues worldwide, but in Ecuador, reports on the microbiological quality of food are scarce. In this cross-sectional study, 450 samples of high-demand Ecuadorian food, including bolon, encebollado, sauces, ceviche, fruit, fruit juice, fruit salad, cheese, raw chicken, and ground beef, were collected from popular street markets in the cities of Guayaquil, Quito, and Cuenca. Populations of total aerobic mesophilic bacteria, total coliforms, fecal coliforms, Escherichia coli, Salmonella enterica, and Listeria monocytogenes were examined on composited samples by plate count following the local regulations (Norma Tecnica Ecuatoriana, Instituto Ecuatoriano de Normalización) for each kind of food. The individual and interaction effects of the city and food type on the levels of each bacterial group were assessed by two-way analysis of variance. Selected colonies from each culture were identified using Biolog OmniLog ID and sequencing of the V3 to V4 region on the 16S rRNA gene. Average total aerobic mesophilic bacteria, total coliform, fecal coliform, and E. coli levels were 5.10 ± 0.12, 2.50 ± 0.16, 1.09 ± 0.12, and 0.83 ± 0.12 log CFU/g or mL, respectively, with significant variations among the cities. The prevalence of Salmonella in chicken and sauces and L. monocytogenes in cheese and fruit salad was greater than 20%. Opportunistic pathogens including Klebsiella pneumoniae, Staphylococcus sciuri, and Enterococcus spp. were frequently identified in the samples from all three cities. High prevalence of spoilage microorganisms such as Bacillus amyloliquefaciens and biocontrol bacteria such as Lactococcus lactis was also observed. This is the first report on the microbiological quality of food from Ecuador.
Subject(s)
Food Microbiology , Listeria monocytogenes , Animals , Cattle , Cities , Colony Count, Microbial , Cross-Sectional Studies , Ecuador , Escherichia coli , RNA, Ribosomal, 16S , StaphylococcusABSTRACT
Outbreaks of Salmonella enterica have been associated with water sources. Survival of S. enterica in various environments has been studied but survival in water has rarely been attempted. In two separate experiments, we examined the survival of S. enterica Typhimurium in clean spring water at various eutrophication levels and temperatures. In the first experiment, lasting for 135 days, survival of S. enterica (10(10) CFU/ml) in water with 0, 50, 100, 500, and 1,000 mg/liter of added carbon at 7, 17, and 27°C was monitored weekly. In the second experiment, lasting for 3 weeks, survival of S. enterica in water at 0, 100, and 200 mg/ liter of added carbon and 27°C was studied daily. Each experiment had four replicates. Dissolved organic carbon was measured daily in each experiment. At the beginning, midpoint, and end of the survival study, microbial communities in both experiments were assessed by denaturing gradient gel electrophoresis (DGGE). Even at minimal carbon concentrations, S. enterica survived for at least 63 d. Survival of Salmonella was highly dependent on eutrophication levels (as measured by dissolved organic carbon) and temperature, increasing at high eutrophication levels, but decreasing at high temperatures. Survival was also strongly affected by microbial competition or predation.