Presence and correlation of some enteric indicator bacteria, diarrheagenic Escherichia coli pathotypes, and Salmonella serotypes in alfalfa sprouts from local retail markets in Pachuca, Mexico.

Data on the presence of diarrheagenic Escherichia coli pathotypes (DEPs) in alfalfa sprouts and correlations between the presence of coliform bacteria (CB), fecal coliforms (FC), E. coli, DEPs, and Salmonella in alfalfa sprouts are not available. The presence of and correlations between CB, FC, E. coli, DEPs, and Salmonella in alfalfa sprouts were determined. One hundred sprout samples were collected from retail markets in Pachuca, Hidalgo State, Mexico. The presence of indicator bacteria and Salmonella was determined using conventional culture procedures. DEPs were identified using two multiplex PCR procedures. One hundred percent of samples were positive for CB, 90% for FC, 84% for E. coli, 10% for DEPs, and 4% for Salmonella. The populations of CB ranged from 6.2 up to 8.6 log CFU/g. The FC and E. coli concentrations were between , 3 and 1,100 most probable number (MPN)/g. The DEPs identified included enterotoxigenic E. coli (ETEC; 2%), enteropathogenic E. coli (EPEC; 3%), and Shiga toxin-producing E. coli (STEC; 5%). No E. coli O157:H7 strains were detected in any STEC-positive samples. In samples positive for DEPs, the concentrations ranged from 210 to 240 MPN/g for ETEC, 28 to 1,100 MPN/g for EPEC, and 3.6 to 460 MPN/g for STEC. The Salmonella isolates identified included Salmonella enterica serotype Typhimurium in three samples and Salmonella enterica serotype Enteritidis in one. STEC and Salmonella Typhimurium were identified together in one sample. Positive correlations were observed between FC and E. coli, between FC and DEPs, and between E. coli and DEPs. Negative correlations occurred between CB and DEPs and between CB and Salmonella. Neither FC nor E. coli correlated with Salmonella in the sprout samples. To our knowledge, this is the first report of ETEC, EPEC, and STEC isolated from alfalfa sprouts and the first report of correlations between different indicator groups versus DEPs and Salmonella.

Presence of Shiga toxin-producing Escherichia coli, Enteroinvasive E. coli, Enteropathogenic E. coli, and Enterotoxigenic E. coli on tomatoes from public markets in Mexico.

Diarrheagenic Escherichia coli pathotypes (DEP) are important foodborne pathogens in various countries, including Mexico. However, no data exist on the presence of DEP on fresh tomatoes (Solanum lycopericum) from Mexico. The frequency of fecal coliforms (FC), E. coli, and DEP were determined for two tomato varieties. One hundred samples of a saladette tomato variety and 100 samples of a red round tomato variety were collected from public markets in Pachuca, Mexico. Each tomato sample consisted of four whole tomatoes. For the 100 saladette samples, coliform bacterial, FC, E. coli, and DEP were identified in 100, 70, 60, and 10% of samples, respectively. For the 100 red round samples, coliform bacterial, FC, E. coli, and DEP were identified in 100, 75, 65, and 11% of samples, respectively. Identified DEP included Shiga toxin-producing E. coli (STEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), and enterotoxigenic E. coli (ETEC). STEC were isolated from 6% of saladette samples and 5% of red round samples. ETEC were isolated from 3% of saladette samples and 4% of red round samples. EPEC were isolated from 2% of saladette samples and 3% of red round samples, and EIEC were isolated from 1% of saladette samples. Both STEC and ETEC were identified in two saladette samples and 1 red round sample. E. coli O157:H7 was not detected in any STEC-positive samples.

Behavior of non-O157 Shiga toxin-producing Escherichia coli, enteroinvasive E. coli, enteropathogenic E. coli, and enterotoxigenic E. coli strains on alfalfa sprouts.

Data about the behavior of non-O157 Shiga toxin-producing Escherichia coli (non-O157 STEC), enteroinvasive E. coli (EIEC), enterotoxigenic E. coli (ETEC), and enteropathogenic E. coli (EPEC) on seeds and alfalfa sprouts are not available. The behavior of STEC, EIEC, ETEC, and EPEC was determined during germination and sprouting of alfalfa seeds at 20 ± 2°C and 30 ± 2°C and on alfalfa sprouts at 3 ± 2°C. When alfalfa seeds were inoculated with STEC, EIEC, ETEC, or EPEC strains, all these diarrheagenic E. coli pathotypes (DEPs) grew during germination and sprouting of seeds, reaching counts of approximately 5 and 6 log CFU/g after 1 day at 20 ± 2°C and 30 ± 2°C, respectively. However, when the sprouts were inoculated after 1 day of seed germination and stored at 20 ± 2°C or 30 ± 2°C, no growth was observed for any DEP during sprouting at 20 ± 2°C or 30 ± 2°C for 9 days. Refrigeration reduced significantly (P < 0.0.5) the number of viable DEPs on sprouts after 20 days in storage; nevertheless, these decreases have no practical significance for the safety of the sprouts.

Escherichia coli O157 in ground beef from local retail markets in Pachuca, Mexico.

Escherichia coli O157 strains have been recognized as pathogenic bacteria, of which raw beef is a known vehicle. An evaluation was done of the presence of E. coli O157 in ground beef from local retail markets in Pachuca, Hidalgo State, Mexico. A total of 120 ground beef samples (500 g) were tested for E. coli O157 by simultaneous application of the U. S. Department of Agriculture, Food Safety and Inspection Service (FSIS)'s Microbiology Laboratory Guidebook culture procedure 5.05, and two commercial kits, Reveal for E. coli O157:H7 and Visual Immunoprecipitate Assay (VIP) Gold for enterohemorrhagic E. coli. Two incubation times (8 and 20 h) were used with the commercial kits. Presence of stx1, stx2, and eaeA loci was determined by multiplex PCR. Of 360 subsamples (120 per procedure), 12 samples were found to be E. coli O157 positive by the FSIS culture method. With VIP, 73 subsamples were presumptive positive after 8 h of enrichment, and 60 were presumptive positive after 20 h of enrichment. Of these, only 6 (8 h) and 8 (20 h) subsamples were confirmed true positives with the FSIS method. With Reveal, 60 subsamples were presumptive positive after 8 h of enrichment and 50 were presumptive positive after 20 h of enrichment. Of these, only 6 (8 h) and 8 (20 h) subsamples were confirmed as true positives with the FSIS method. A total of 57 E. coli O157:H7 and 21 E. coli O157 strains were isolated. None of the O157 or O157:H7 strains had stx1 or stx2 loci, and only one had the eaeA locus. To our knowledge, this is the first report of the presence of E. coli O157 in commercial ground beef from Mexico, and the first report of isolation of a large number of stx-negative E. coli O157 and E. coli O157:H7 strains in Mexico.

Presence of some indicator bacteria and diarrheagenic E. coli pathotypes on jalapeño and serrano peppers from popular markets in Pachuca City, Mexico.

The chili pepper is a very important crop in Mexico. Diarrheagenic E. coli pathotypes (DEPs) are important foodborne pathogens in different countries including Mexico. No data exists on DEPs presence on fresh jalapeño and serrano pepper and little data have been published on the microbiological quality of these peppers. The frequencies of coliform bacteria (CB), thermotolerant coliforms (TC), E. coli and DEPs were determined for jalapeño and serrano peppers. Of 100 serrano samples, CB, TC, E. coli and DEPs were identified in 100, 90, 58 and 36%, respectively. Of 100 jalapeño samples, CB, TC, E. coli and DEPs were identified in 100, 88, 38 and 14%, respectively. Identified DEPs included enterotoxigenic E. coli (ETEC) and Shiga toxin-producing E. coli (STEC). STEC were isolated from 36% of serrano samples and 14% of jalapeño samples. ETEC were isolated from 12% of serrano samples and 2% of jalapeño samples. Both STEC and ETEC were identified in 14 serrano samples and 2 jalapeño samples. No E. coli O157:H7 were detected in any STEC-positive samples. Jalapeño and serrano peppers could be an important factor contributing to the endemicity of DEPs-caused gastroenteritis in Mexico.

Frequency and behavior of Salmonella and Escherichia coli on whole and sliced jalapeño and serrano peppers.

The frequencies of coliform bacteria (CB), thermotolerant coliforms (TC), Escherichia coli, and Salmonella were determined for jalapeño and serrano peppers. In addition, the behavior of four serotypes of Salmonella and three E. coli strains on whole and sliced jalapeño and serrano peppers as well as in blended sauce at 25 ± 2°C and 3 to 5°C was investigated. Chili peppers were collected from markets in the city of Pachuca, Hidalgo, Mexico. CB, TC, E. coli, and Salmonella were detected on serrano peppers in 100, 90, 50, and 10 % of the samples, and on jalapeño peppers in 100, 86, 32, and 12 % of the samples. Concentrations of CB ranged from 3.8 to 7.9 log CFU per serrano sample and from 5.3 to 8.2 log CFU per jalapeño sample, whereas concentrations of TC and E. coli were between < 3 and 1,100 most probable number per serrano and jalapeño samples. On whole serrano and jalapeño peppers stored at 25 ± 2°C or 3 to 5°C, no growth was observed for rifampin-resistant strains of Salmonella and E. coli. After 6 days at 25 ± 2°C, the tested Salmonella serotypes and E. coli strains had decreased from an initial inoculum level of 5 log CFU to 1 and 2.5 log on serrano and jalapeño peppers, respectively, and at 3 to 5°C they decreased to approximately 1.8 and 1.2 log, respectively, on serrano and jalapeño. Both the Salmonella serotypes and E. coli grew on sliced chili peppers and in blended sauce; after 24 h at 25 ± 2°C, both bacteria types had grown to approximately 4 and 5 log CFU on pepper slices and in sauce, respectively. At 3 to 5°C the bacterial growth was inhibited.

Behavior of Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Shigella flexneri and Shigella sonnei during production of pulque, a traditional Mexican beverage.

Pulque is a typical fermented alcoholic beverage of central Mexico, produced from the nectar of maguey agave plants. Production systems are largely artisanal, with inadequate hygiene conditions and exposure to multiple contamination sources. No data exist on pulque microbiological safety and the behavior of pathogenic microorganisms in agave nectar and pulque. An initial trial was done of the behavior of Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Shigella flexneri and Shigella sonnei during fermentation of nectar from a single producer, nectar mixture from different producers, and seed pulque. A second trial simulating artisanal pulque production was done by contaminating fresh nectar with each of the five strains, storing at 22°C for 14 h, adding seed pulque, and fermenting until pulque was formed. During incubation at 16 or 22°C in the first trial, all the pathogenic strains multiplied in both the single producer nectar and the nectar mixture, reaching maximum concentrations at 12 h. Strains concentration then decreased slowly. In the seed pulque, the strains did not multiply and tended to die. In the second trial, all strains increased concentration from 0.7 to 1.6 log at 22°C, and from 0.5 to 1.1 at 16°C in the first 14 h. After addition of seed pulque, they were quickly deactivated until none was detected in the final product. The results suggest that the potential risk to consumers of contracting any of the five tested pathogenic bacterial strains from pulque is low.