Ethanol inhibits dopamine uptake via organic cation transporter 3: Implications for ethanol and cocaine co-abuse.

Concurrent cocaine and alcohol use is among the most frequent drug combination, and among the most dangerous in terms of deleterious outcomes. Cocaine increases extracellular monoamines by blocking dopamine (DA), norepinephrine (NE) and serotonin (5-HT) transporters (DAT, NET and SERT, respectively). Likewise, ethanol also increases extracellular monoamines, however evidence suggests that ethanol does so independently of DAT, NET and SERT. Organic cation transporter 3 (OCT3) is an emergent key player in the regulation of monoamine signaling. Using a battery of in vitro, in vivo electrochemical, and behavioral approaches, as well as wild-type and constitutive OCT3 knockout mice, we show that ethanol's actions to inhibit monoamine uptake are dependent on OCT3. These findings provide a novel mechanistic basis whereby ethanol enhances the neurochemical and behavioral effects of cocaine and encourage further research into OCT3 as a target for therapeutic intervention in the treatment of ethanol and ethanol/cocaine use disorders.

Hydrophobic and adhesive patterns of lactic acid bacteria and their antagonism against foodborne pathogens on tomato surface (Solanum lycopersicum L.).

AIMS: To evaluate tomato epiphyte lactic acid bacteria (LAB) hydrophobicity and auto-aggregation as an indicator of bacteria adhesion to tomato. Likewise, use LAB adhesion and co-aggregation as mechanisms to antagonize pathogen attachment. METHODS AND RESULTS: Fifty-four LAB were screened to evaluate their hydrophobic, auto- and co-aggregative properties against Salmonella Typhimurium, Saintpaul, Montevideo and Escherichia coli O157:H7. Subsequently, tomato adhesion of Enterococcus faecium Col1-1C, Weisella cibaria 11-E-2 and W. confusa Col 1-13 with high, medium and low hydrophobicity and high co-aggregation was investigated as well as their pathogen antagonism. Results indicate that bacteria hydrophobicity and auto-aggregation correspond to LAB adhesion to tomato. Enterococcus faecium Col1-1C interfered in most of the pathogen adhesion and micrographs revealed that such effect could be related to the inhibition of structures-type biofilm on E. coli O157:H7 and the aggregate formation on Salmonella. CONCLUSIONS: Lactic acid bacteria hydrophobicity and auto-aggregation can estimate bacteria adhesion to tomato and adhesive and co-aggregative properties could serve as a tool to antagonize foodborne pathogens under specific conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study evidence the interference of Ent. faecium Col1-1C in E. coli O157:H7 biofilm formation and Salmonella colonization.

Prevalence and behavior of multidrug-resistant Salmonella strains on raw whole and cut nopalitos (Opuntia ficus-indica L.) and on nopalitos salads.

BACKGROUND: The presence of multidrug-resistant Salmonella in vegetables is a significant public health concern. Nopalito is a cactaceous that is commonly consumed either raw or cooked in Mexico and other countries. The presence of antibiotic-resistant Salmonella strains on raw whole nopalitos (RWN, without prickles), raw nopalitos cut into squares (RNCS) and in cooked nopalitos salads (CNS) samples was determined. In addition, the behavior of multidrug-resistant Salmonella isolates on RWN, RNCS and CNS at 25° ± 2 °C and 3° ± 2 °C was investigated. RESULTS: One hundred samples of RWN, 100 of RNCS and 100 more of CNS were collected from public markets. Salmonella strains were isolated and identified in 30, 30 and 10% of the samples, respectively. Seventy multidrug-resistant Salmonella strains were isolated from all the nopalitos samples. Multidrug-resistant Salmonella isolates survived at least 15 days on RWN at 25° ± 2 °C or 3° ± 2 °C. Multidrug-resistant Salmonella isolates grew in the RNCS and CNS samples at 25° ± 2 °C. However, at 3° ± 2 °C the bacterial growth was inhibited. CONCLUSION: This is the first report about multidrug-resistant Salmonella isolation from raw nopalitos and nopalitos salads. Nopalitos from markets are very likely to be an important factor contributing to the endemicity of multidrug-resistant Salmonella-related gastroenteritis in Mexico. © 2017 Society of Chemical Industry.

Behaviour of four diarrheagenic Escherichia coli pathotypes on carrots and in unpasteurized carrot juice.

UNLABELLED: The behaviours of Shiga toxin-producing Escherichia coli (STEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC) strains on raw carrots at 3 ± 1 and 30 ± 1°C, and in unpasteurized carrot juice at 3 ± 1, 12 ± 1, 20 ± 1, 30 ± 1°C and 37 ± 1°C were determined. Raw carrots were purchased in a local market. Fresh juice was obtained from raw carrots in the laboratory. On whole carrots stored at 30 ± 1 or 3 ± 1°C, no growth was observed for any of the diarrheagenic E. coli pathotype (DEPs) strains studied. After 15 days at 30 ± 1°C, the tested DEPs had decreased from an initial inoculum level of approximately 6 log colony-forming units (CFU) to approximately 3·5 log CFU on whole carrots, while at 3 ± 1°C, they decreased from approximately 2·4 log to 1·6 log CFU. All these DEPs grew in fresh carrot juice at 12 ± 1, 20 ± 1, 30 ± 1 and 37 ± 1°C, reaching counts of approximately 4·2 log, 5·8 log, 6·7 log and 7·5 log CFU ml(-1) , respectively, after 24 h. At 3 ± 1°C, the DEP growth was inhibited at least during 7 days. Thus, storage of carrot juice at unrefrigerated temperatures can result in DEP growth to levels likely to represent a risk to consumers. SIGNIFICANCE AND IMPACT OF THE STUDY: The information presented shows the potential of Shiga toxin-producing Escherichia coli, enteroinvasive E. coli, enteropathogenic E. coli and enterotoxigenic E. coli strains for survival on carrots and growth in carrot juice at warmer temperatures. The information can help food processors in plants and restaurants understand the importance of the implementation of hazard analysis and critical control point (HACCP) strategies for preventing potential diarrheagenic E. coli pathotypes (DEPs) contamination and growth in carrot juice. This is the first report regarding the behaviour these DEPs on carrots and in carrot juice.

Presence of indicator bacteria, diarrhoeagenic Escherichia coli pathotypes and Salmonella in fresh carrot juice from Mexican restaurants.

UNLABELLED: Coliform bacteria (CB), faecal coliforms (FC), Escherichia coli, diarrhoeagenic E. coli pathotypes (DEP) and Salmonella frequencies were determined for fresh carrot juice from restaurants in Pachuca city, Mexico. Two hundred and eighty carrot juice samples were purchased in three types of restaurants: (A), national chain restaurants; (B), local restaurants; and (C), very small restaurants. Two restaurants for each A and B, and three for C, were included. Forty juice samples were purchased at each restaurant. All tested juice samples had poor microbiological quality. Of these samples, 100, 96·8, 54·3, 8·9 and 8·6% had CB, FC, E. coli, DEP and Salmonella, respectively. CB were present in all juice samples regardless of source, with limits ranging from 3·6 × 10² to 8·5 × 107 CFU ml⁻¹, and the limits for FC and E. coli were <3 to 1100 MPN ml⁻¹ and <3 to 460 MPN, respectively. DEP and Salmonella were isolated from samples from all the restaurants at levels of 5% or above: DEP, 5% (A1, B2, 10% (A2, B1, C1, C2) and 12·5% (C3); Salmonella, 5% (A1, A2, B2), 7·5% (C2), 10% (C1, 12·5% (B1) and 15% (C3). SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of microbiological quality and Salmonella, enteroinvasive E. coli (EIEC), enterotoxigenic E. coli (ETEC) and Shiga toxin-producing E. coli (STEC) isolation from fresh carrot juice in Mexico. Fresh carrot juice from restaurants could be an important factor contributing to the endemicity of EIEC-, ETEC- and STEC- and Salmonella-caused gastroenteritis 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.

Incidence and behavior of Salmonella and Escherichia coli on whole and sliced zucchini squash (Cucurbitapepo) fruit.

The incidence of coliform bacteria (CB), thermotolerant coliforms (TC), Escherichia coli, and Salmonella was determined for zucchini squash fruit. In addition, the behavior of four serotypes of Salmonella and a cocktail of three E. coli strains on whole and sliced zucchini squash at 25+/-2 degrees C and 3 to 5 degrees C was tested. Squash fruit was collected in the markets of Pachuca city, Hidalgo State, Mexico. CB, TC, E. coli, and Salmonella were detected in 100, 70, 62, and 10% of the produce, respectively. The concentration ranged from 3.8 to 7.4 log CFU per sample for CB, and >3 to 1,100 most probable number per sample for TC and E. coli. On whole fruit stored at 25+/-2 degrees C or 3 to 5 degrees C, no growth was observed for any of the tested microorganisms or cocktails thereof. After 15 days at 25+/-2 degrees C, the tested Salmonella serotypes had decreased from an initial inoculum level of 7 log CFU to <1 log, and at 3 to 5 degrees C they decreased to approximately 2 log. Survival of E. coli was significantly greater than for the Salmonella strains at the same times and temperatures; after 15 days, at 25+/-2 degrees C E. coli cocktail strains had decreased to 3.4 log CFU per fruit and at 3 to 5 degrees C they decreased to 3.6 log CFU per fruit. Both the Salmonella serotypes and E. coli strains grew when inoculated onto sliced squash: after 24 h at 25+/-2 degrees C, both bacteria had grown to approximately 6.5 log CFU per slice. At 3 to 5 degrees C, the bacterial growth was inhibited. The squash may be an important factor contributing to the endemicity of Salmonella in Mexico.

Evaluation of the resistance to bacterial growth of star-shaped poly(ε-caprolactone)-co-poly(ethylene glycol) grafted onto functionalized carbon nanotubes nanocomposites.

Nanocomposites of functionalized carbon nanotubes (CNTsf) as nanofillers, and a copolymer of star-shaped poly(ε-caprolactone) (stPCL) and poly(ethylene glycol) (PEG) as a polymeric matrix were synthesized, characterized, and their resistance to the growth of Staphylococcus aureus and Pseudomonas aeruginosa was evaluated. CNTsf contain hydroxyl, carboxyl and acyl chloride groups attached to their surface. Nanocomposites were prepared by mixing CNTsf to a solution of stPCL-PEG copolymer. Raman and FT-IR spectroscopies confirm the functionalization of carbon nanotubes (CNTs). Star-shaped PCL-PEG copolymer was characterized by Gel permeation chromatography (GPC), and 1H-NMR and 13C-NMR spectroscopies. X-ray photoelectron spectroscopy (XPS) shows that CNTsf are grafted to the stPCL-PEG copolymer. Crystallization behavior of the nanocomposites depends on the amount of CNTsf used in their preparation, detecting nucleation (nanocomposites prepared with 0.5 wt.% of CNTsf) or anti-nucleation (nanocomposites prepared with 1.0 wt.% of CNTsf) effects. Young's Moduli and thermal stability of nanocomposites were higher, but their resistence to the proliferation of Staphylococcus aureus and Pseudomonas aeruginosa was lower than the observed for their pure polymer matrix.

Isolation and characterization of Shiga toxin-producing Escherichia coli O157:H7 and non-O157 from beef carcasses at a slaughter plant in Mexico.

The contamination of beef carcasses with Shiga toxin-producing O157:H7 and non-O157 Escherichia coli (STEC) obtained from a slaughter plant in Guadalajara, Mexico was investigated. A total of 258 beef carcasses were sampled during a 12-month period. All samples were assayed for STEC by selective enrichment in modified tryptone soy broth supplemented with cefixime, cefsulodin and vancomycin, followed by plating on Sorbitol MacConkey Agar supplemented with cefixime and tellurite (CT-SMAC). Simultaneously, all samples were assayed by immunomagnetic separation (IMS) and plated on CT-SMAC and CHROMagar. The presence of the stx1, stx2, eaeA and hly933 genes, recognized as major virulence factors of STEC, was tested for O157:H7 and non-O157 E. coli isolates by multiplex polymerase chain reaction (PCR). STEC was detected in two (0.8%) samples. One of these STEC isolates corresponded to the serotype O157:H7 showing stx2, eaeA and hyl933 genes. The other isolate corresponded to non-O157 STEC and only had the stx1 gene. Thirteen carcasses (5%) were positive for nonmotile E. coli O157 and 7 (2.7%) were positive for E. coli O157:H7. The presence of O157:H7 and non-O157 STEC on beef carcasses in this slaughter plant in Guadalajara, Mexico, emphasizes the importance of implementing the Hazard Analysis and Critical Control Point (HACCP) system, as well as the need for implementing, evaluating, and validating antimicrobial interventions to reduce the presence of potential pathogenic microorganisms.

Salmonella and Shigella in freshly squeezed orange juice, fresh oranges, and wiping cloths collected from public markets and street booths in Guadalajara, Mexico: incidence and comparison of analytical routes.

A survey of the presence of Salmonella and Shigella in freshly squeezed orange juice and related samples was conducted in Guadalajara, Mexico. One hundred samples of freshly squeezed orange juice were collected from 49 street booths and 51 small food service establishments. In addition, 75 fresh orange samples, each consisting of five orange units, and 75 wiping cloths were collected from the same establishments from which juice had been collected. Salmonella was isolated from 14, 20, and 23% of samples of orange juice, orange surfaces, and wiping cloths collected from street vendors, while Shigella was isolated from 6, 17, and 5% of these samples. In general, the frequency of isolation of these pathogens in samples from juice serving establishments at public markets was significantly lower than that found among street vendors (P < 0.05). Salmonella enterica serotypes Agona, Typhimurium, and Anatum were found in orange juice, fresh oranges, and wiping cloth samples, while serotype Mexico was found on fresh oranges and in wiping cloths and serotypes Muenchen and Panama were found only in wiping cloth samples. Regarding Shigella species, Shigella sonnei was found in all three types of sample tested; Shigella dysenteriae was found in juice and orange samples, Shigella boydii in orange and wiping cloth samples, and Shigella flexneri on oranges only. Thirty-one percent and 39% of the juice samples showed aerobic plate counts of > or = 5.0 log CFU/ml and Escherichia coli counts of > 3.0 log CFU/ml, respectively. These high counts may indicate poor sanitation and potential exposure to fecal contamination either in the raw materials or during the orange-crushing and juice-serving process. These data may be useful for a further risk assessment of Salmonella or Shigella in unpasteurized, freshly squeezed juice.

Behavior and Inactivation of Enterotoxin-Positive Clostridium perfringens in Pork Picadillo and Tamales Filled with Pork Picadillo under Different Cooking, Storage, and Reheating Conditions.

This study analyzed the behavior of Clostridium perfringens in individual ingredients and tamales containing different pathogen concentrations upon exposure to different temperatures and methods of cooking, storage, and reheating. In ground pork, C. perfringens cells were inactivated when exposed to 95°C for 30 min. Three lots of picadillo inoculated with 0, 3, and 5 log CFU/g C. perfringens cells, respectively, were exposed to different storage temperatures. At 20°C, cell counts increased 1 log in all lots, whereas at 8°C, counts decreased by 2 log. Four lots of tamales prepared with picadillo inoculated with 0, 2, 3, and 7 log CFU/g prior to the final cooking step exhibited no surviving cells (91°C for 90, 45, or 35 min). Four lots of tamales were inoculated after cooking with concentrations of 0, 0.6, 4, and 6 log CFU/g of the pathogen and then stored at different temperatures. In these preparations, after 24 h at 20°C, the count increased by 1.4, 1.7, and 1.8 log in the tamales inoculated with 0.6, 4, and 6 log inoculum, respectively. When they were stored at 8°C for 24 h, enumerations decreased to <1, 2.5, and 1.9 log in the tamales inoculated with 0.6, 4, and 6 log of C. perfringens cells, respectively. However, when the lots were exposed to 20°C and then 8°C, 0.8, 1.8, and 2.4 log changes were observed for the tamales inoculated with 0.6, 4, and 6 log, respectively. Microwaving, steaming, and frying to reheat tamales inoculated with 6 log CFU/g C. perfringens cells showed that the pathogen was inactivated after 2 min of exposure in the microwave and after 5 min of exposure to steam. In contrast, no inactivation was observed after 5 min of frying. The tamales inoculated with spores (7 log most probable number [MPN]/g) showed a decrease of 2 log after steaming or frying, and no survival was observed after microwaving. Tamales inoculated with spores (7 log MPN/g) after cooking were susceptible to microwaves, but 2.4 and 255 MPN/g remained after frying and steaming, respectively.

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.

Enhanced gamma-aminobutyric acid-B receptor agonist responses and mRNA within the nucleus of the solitary tract in hypertension.

Gamma-Aminobutyric acid-B (GABAB) receptor function and regulation in the nucleus of the solitary tract (NTS) was examined in Sprague-Dawley rats made chronically (4 to 5 weeks) hypertensive with the one-kidney, figure-8 renal wrap model of hypertension. NTS microinjection of the GABAB agonist baclofen produced a pressor response that was enhanced in hypertensive rats compared with the response observed in sham-operated normotensive rats (36+/-4 mm Hg increase in mean arterial pressure in 8 hypertensive rats compared with 21+/-2 mm Hg increase in 7 sham-operated normotensive rats, P=0. 03). Responses to microinjection of GABAB antagonists (CGP-55845A and SCH-90511), the GABAA agonist muscimol, the GABAA antagonist bicuculline, and the GABA reuptake inhibitor nipecotic acid were not different comparing normotensive sham-operated and hypertensive rats. Renal sympathetic nerve responses to NTS microinjection of these drugs were not different in hypertensive compared with normotensive rats. Micropunches of the NTS were homogenized and reverse transcriptase-polymerase chain reaction was performed to examine mRNA levels for the GABAB receptor. There was a 3-fold increase in GABAB receptor mRNA levels in the caudal NTS of 7 chronically hypertensive rats compared with levels measured in 8 sham-operated normotensive rats (P=0.01). In conclusion, chronic hypertension is associated with an upregulation of GABAB receptor function; however, the tonic activity of the system does not appear to be different between normotensive and hypertensive rats. The upregulation of GABAB receptor function might be due to an increased number of receptors, as suggested by the elevated levels of GABAB receptor mRNA measured in the NTS of hypertensive rats. All of these alterations suggest that hypertension is associated with dynamic changes in receptor-mediated mechanisms within the NTS, and these alterations could modify baroreflex regulation of cardiovascular function in hypertension.

Spread of bacterial pathogens during preparation of freshly squeezed orange juice.

To study the potential of three bacterial pathogens to cross-contaminate orange juice during extraction, normal operation conditions during juice preparation at food service establishments were simulated. The spread of Salmonella enterica serovar Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes from inoculated oranges to work surfaces and to the final product was determined. The transference of these three bacterial pathogens to orange juice made from uninoculated oranges with the use of contaminated utensils was also studied. Fresh oranges were inoculated with a marker strain of rifampicin-resistant Salmonella Typhimurium, E. coli O157:H7, or L. monocytogenes. Final pathogen levels in juice were compared as a function of the use of electric or mechanical juice extractors to squeeze orange juice from inoculated oranges. Pathogen populations on different contact surfaces during orange juice extraction were determined on sulfite-phenol red-rifampicin plates for Salmonella Typhimurium and E. coli O157:H7 and on tryptic soy agar supplemented with 0.1 g of rifampicin per liter for L. monocytogenes. After inoculation, the average pathogen counts for the orange rind surface were 2.3 log10 CFU/cm2 for Salmonella Typhimurium, 3.6 log10 CFU/cm2 for E. coli O157:H7, and 4.4 log10 CFU/cm2 for L. monocytogenes. This contamination was spread over all utensils used in orange juice squeezing. Mean pathogen counts for the cutting board, the knife, and the extractor ranged from -0.3 to 2.1 log10 CFU/cm2, and the juice contained 1.0 log10 CFU of Salmonella Typhimurium per ml, 2.3 log10 CFU of E. coli O157:H7 per ml, and 2.7 log10 CFU of L. monocytogenes per ml. Contact with contaminated surfaces resulted in the presence of all pathogens in orange juice made from uninoculated oranges. These results give emphasis to the importance of fresh oranges as a source of pathogens in orange juice.

Survival of Vibrio cholerae O1 in ceviche and its reduction by heat pretreatment of raw ingredients.

The survival of Vibrio cholerae O1 serotypes Inaba and Ogawa was determined in ceviche prepared from inoculated ground fish. Ground mackerel purchased from a seafood distribution center was inoculated with V. cholerae and stored at 8 or 20 degrees C. Counts of V. cholerae decreased in 2.6 to 2.7 log10 CFU/g during 96 h of storage at 8 degrees C or 2.5 to 2.6 log10 CFU/g during 24 h at 20 degrees C. Survival studies indicated that serotype Inaba decreased its number following a linear or retarded trend, whereas serotype Ogawa followed an accelerated death trend. No effect of the initial level of inoculum was observed. Odor scores of ceviche indicated that this food became marginally acceptable within as little as 48 h of storage at 8 degrees C or 3 h at 20 degrees C and were related to total volatile nitrogen values but not to aerobic plate counts, pH, or coliform counts. A heat pretreatment that consisted of stirring 100 g of inoculated ground fish into 40 ml of boiling water produced an 8-log reduction of V. cholerae within 3 min without affecting the color, odor, or flavor of ceviche prepared with such pretreated fish. According to this study, V. cholerae present in contaminated ceviche will likely survive longer than the shelf life of this food. Preheating the ground raw fish used for preparing ceviche for 3 min should effectively eliminate V. cholerae O1, providing science-based conditions for implementing a critical control point if a hazard analysis critical control point plan were to be developed for preparation of ceviche.

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.

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.

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.

Frequency of Salmonella and Listeria monocytogenes in five commercial brands of chicken eggs using a combined method of enrichment and nested-PCR.

Eggs or egg-based foods, either raw or undercooked, have been identified as vehicles of Salmonella outbreaks. The low numbers of Salmonella organisms in eggs makes it difficult to detect them in frequency studies. The nested-PCR (n-PCR) technique shows more sensitivity and specificity than bacteriological culture methods (BCMs). A preenrichment method followed by enrichment and n-PCR is a good alternative for the investigation of Salmonella and Listeria monocytogenes in eggs. A total of 2,650 chicken eggs representing five commercial brands were purchased from 10 grocery stores. Ten eggs of each brand were combined in order to obtain 265 pooled samples (53 per brand). The shells and yolks of 100 pooled samples were analyzed for Salmonella, while the shells of 65 pooled samples were analyzed for L. monocytogenes, using BCM and a combined method of enrichment and n-PCR (CM-n-PCR). Sixteen eggshell pooled samples tested positive for Salmonella by CM-n-PCR, compared with only two by BCM. Three egg yolk pooled samples tested positive for this pathogen by CM-n-PCR; none tested positive by BCM. Three eggshell pooled samples tested positive for L. monocytogenes by CM-n-PCR and none by BCM. In Mexico, as in other countries, official methods for detection of Salmonella and L. monocytogenes in foods are based on standard bacteriological culture techniques. The inclusion of more sensitive methods such as the one used in the present investigation would increase the probability of detecting positive samples, particularly in those foods in which a very low number of cells is expected.

Microbiological safety of domestic refrigerators and the dishcloths used to clean them in Guadalajara, Jalisco, Mexico.

Household refrigerators are a potential pathogen contamination source for foods. An evaluation of the microbiological safety of 200 refrigerators in Guadalajara, Jalisco, Mexico, was made by visual inspection, ATP-bioluminescence levels, indicator microorganisms including Escherichia coli and Staphylococcus aureus, and the presence of Listeria monocytogenes and Salmonella. Additionally, interviews of the owners of the refrigerators were carried out to determine relationships between food storage practices, demographic aspects, and microbiological status. Dishcloths used to clean refrigerators were also analyzed. Operational conditions (cleanliness, fullness, organization, frequency of cleaning, and temperature) were evaluated by trained observers. Results showed deficient cleanliness in 55% of refrigerators, 22% were completely full, 43% very disorganized, 28% were usually cleaned only once in 3 to 6 months, and 53% had internal temperatures >7.1°C. ATP-bioluminescence levels were >300 relative light units on 67 and 74% of shelves and drawers, respectively, indicating that surfaces were dirty according to the luminometer manufacturer. Psychrotrophic aerobic bacteria counts on shelves, drawers, and dishcloths were 6.3, 5.2, and 6.3 log CFU/cm(2); for coliform bacteria, 5.2, 3.9, and 4.7 CFU/cm(2); for E. coli, 3.7, 3.5, and 4.8 CFU/cm(2); and for Staphylococcus aureus, 2.1, 2.5, and 2.3 CFU/cm(2), respectively. L. monocytogenes and Salmonella were isolated from 59.5, 20.5, and 17% and 32.5, 8.0 and 12.5% of shelves, drawers, and dishcloths, respectively. Four Salmonella serotypes and nine serogroups (partially serotyped isolates) were identified. The most prevalent were Salmonella Anatum (39.5%), Salmonella group E1 (19.7%), and Salmonella group E1 monophasic (12.5%). Operational conditions and microbiological status were clearly deficient in sampled refrigerators, highlighting the consequent risk of foodborne disease among users. Educational programs are needed to improve the domestic food safety in Mexico.