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There is a lack of information about Salmonella enterica strains under acidic conditions and their association with their genome. This study characterized intraspecies variability in the growth of 167 S. enterica isolates under two acid conditions (pH 4 and 5) and linked to the whole genome sequencing (WGS) data. A total of 1002 curves for each condition were obtained using turbidimetry measurements, and Baranyi and Roberts model was used to estimate the maximum rate of change (rcmax; OD600 nm h-1). Strains were categorized into slow, intermediate, and fast; and associations with their WGS data were performed. Huge variability in r c max ¯ $\overline {{\mathrm{r}}{{{\mathrm{c}}}_{{\mathrm{max}}}}} $ was observed at both conditions (pH 5 = 0.016-0.066 OD600nm h-1 and pH 4 = 0.003-0.028 OD600nm h-1). The majority of isolates was classified as intermediate r c max ¯ $\overline {{\mathrm{r}}{{{\mathrm{c}}}_{{\mathrm{max}}}}} $ (59.5% at pH 5 and 46.1% at pH 4). Strains classified as fast had a low frequency of allABCD genes at both pHs, and any of them having the presence of pefABCD, spvBCR, aadA2, dfrA12, and gyrA_D87G genes were linked to virulence or antimicrobial resistance. This study suggests that strains with fast capacity for growth under acidic conditions could have a fitness cost in their virulence or resistance potential. PRACTICAL APPLICATION: Data presented in this study could be used to select representative strains to evaluate the exposure assessment in different food items, mainly the growth and survival in acidic foods.

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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.

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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.

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The dynamics of bacteria community of "Bola de Ocosingo" cheese, a Mexican artisanal raw milk cheese was investigated by high-throughput sequencing (454 pyrosequencing). Dairy samples (raw milk, curd, cheese at 50 and 110 days of ripening) were collected at dry (March-June) and rainy season (August-November) from three producers located in Chiapas, Mexico. In general, raw milk contained high bacterial diversity which was reduced throughout cheese manufacture. However, in two productions an important increase during cheese ripening was observed probably due to cross-contamination. Species such as Streptococcus thermophilus, Lactococcus lactis, Lactobacillus helveticus, L. delbrueckii and L. plantarum from which potential probiotic strains may be obtained, predominated during processing, varying its prevalence from one producer to another. Furthermore, low proportions of Escherichia coli/Shigella flexnerii were detected in almost all processes, however, could not be recovered by traditional methodology, indicating presence of non-cultivable cells. This work provides insights into bacteria communities of Bola de Ocosingo cheese for starter culture development, many of which are reported to provide health related benefits, and the usefulness of high-throughput sequencing to evidence cross-contamination during processing.

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The bacterial diversity and structure of Poro cheese, an artisanal food, was analysed by high-throughput sequencing (454 pyrosequencing) in order to gain insight about changes in bacterial communities associated with the cheese-making process. Dairy samples consisting of milk, fermented whey, curd and ripened cheese (during 7 and 60 d) were collected from three manufacturers located in the state of Tabasco, México during dry (March-June) and rainy (August-November) seasons. Independently of producer and season, raw milk samples displayed the highest diversity in bacterial communities. In raw milk, genera found were Macrococcus, Staphylococcus, Enterococcus, Streptococcus, Lactobacillus and Enhydrobacter. Diversity in whey, curd and cheese was lower, principally containing Streptococcus and Lactobacillus; however, bacteria such as Staphylococcus, Acinetobacter, Chryseobacterium, Bacillus, Sediminibacter, Lactococcus and Enterococcus were occasionally present. After curdling step, the most dominant and abundant species were Streptococcus thermophilus and Lactobacillus delbrueckii.

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OBJECTIVES: To compare the performance of two rapid systems for the diagnosis of cholera with the culture method, and to propose a strategy for improving the specificity and sensitivity of these systems and reducing the costs involved in making a diagnosis. METHODS: The following institutions participated in the study: the National Bacteriology Referral Center (Centro Nacional de Referencia en Bacteriologia, CNRB) of the Costa Rican Institute for Research and Teaching in Nutrition and Health (Instituto Costarricense de Investigacion y Ensenanza en Nutricion y Salud, INCIENSA) and various hospitals in the provinces of Alajuela, Guanacaste and San Jose, in Costa Rica. A total of 237 feces samples were used to asses the performance of two tests for the rapid detection of Vibrio cholerae 01: the Pathogen Detection Kit (PDK, Intelligent Monitoring Systems, Gainesville, Florida, USA) and Cholera-SMART (New Horizons Diagnostics Corp., Columbia, Maryland, USA), both when applied directly (direct SMART and direct PDK) and when applied to specimens cultured in broth-enriched medium for 6 hours (SMART-6 and CPK-6) and for 18 hours (SMART-18 and PDK-18) at 37 degrees C in alkaline peptone water. Liquid and partially formed stools were cultured and examined by means of the rapid direct test; when the initial result was negative, the tests were repeated after culture for periods of 6 and 18 hours. Rectal and fecal swabs were obtained from feces cultured in enriched-broth medium for 6 and 18 hours. In addition, we studied the sensitivity of the rapid testing systems by using pure cultures of V. cholerae 01 (strain SOS-833, CNRB, Costa Rica) that were incubated for 18 to 24 hours, and we assessed the usefulness of observing motility under the microscope in order to rationalize the use of rapid methods. RESULTS: The sensitivity of the direct SMART test and of the direct PDK test was 100% when samples obtained from liquid and partially formed stools and from the intestinal contents of dead bodies were used. With these samples, the direct SMART procedure showed a specificity of 100%, whereas the direct PDK procedure showed a specificity that ranged from 85.7% to 77.4%, depending on the type of sample. False positives obtained with the direct PDK method turned out to be negative with PDK-6 and PDK-18. Among the rectal and fecal swabs of persons with and without diarrhea or who had received prior treatment with antibiotics, three results that were negative with the SMART-6 procedure and two that were negative with the PDK-6 procedure turned out to be positive with the SMART-18 and PDK-18 procedures, respectively. Both systems showed excellent concordance (kappa index above 0.9) throughout. Both systems were sensitive to 6 x 10(7) colony-forming units per milliliter (cfu/mL), which was concordant with the microscopic observation of 10 microorganisms or more per field with the type of motility that characterizes vibrios (at 1000 x magnification). Samples having fewer than 10 microorganisms with the motility that characterizes vibrios had concentrations between 6 x 10(3) and 6 x 10(6) cfu/mL and became positive only after incubation in enriched-broth medium for 6 to 18 hours. We propose a strategy for diagnosing the presence of V. cholerae 01 infection in less time than it takes with traditional methods, with positive and negative predictive values of 100%. CONCLUSIONS: The SMART and PDK systems make it possible to accurately diagnose cholera quickly, don't require sophisticated equipment or highly qualified technical personnel, and perform satisfactorily in field conditions. Through the proposed strategy, it becomes possible to improve the specificity and sensitivity of these systems and to reduce the cost of making a diagnosis, thus making them suitable for use in cholera surveillance in low-income settings where this disease is a serious public health problem.

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Objetivos. Comparar el desempeño de dos sistemas rápidos de diagnóstico de cólera con el método de cultivo y proponer una estrategia que permita mejorar la especificidad y la sensibilidad de estos sistemas y disminuir los costos del diagnóstico. Métodos. En el estudio participaron el Centro Nacional de Referencia en Bacteriología (CNRB) del Instituto Costarricense de Investigación y Enseñanza en Nutrición y Salud (INCIENSA) y hospitales de las provincias de Alajuela, Guanacaste y San José, en Costa Rica. Se emplearon 237 muestras de heces para evaluar el desempeño de dos pruebas rápidas para el diagnóstico de Vibrio cholerae O1: Pathogen Detection Kit® (PDK, Intelligent Monitoring Systems, Gainsville, Florida, EUA) y Cholera-SMART® (New Horizons Diagnostics Corp., Columbia, Maryland, EUA), tanto en forma directa (SMART directo y PDK directo) como a partir de cultivos de enriquecimiento de 6 horas (SMART-6 y PDK-6) y de 18 horas (SMART18 y PDK-18) a 37 °C en agua de peptona alcalina. Las muestras diarreicas y semiformadas se cultivaron y se evaluaron con las pruebas rápidas directas; cuando el resultado inicial era negativo se repitieron a las 6 y 18 horas de cultivo. Los hisopados rectales y fecales se evaluaron a partir de cultivos de enriquecimiento de 6 y de 18 horas. Adicionalmente se estudió la sensibilidad analítica de los sistemas rápidos con cultivos puros de 18 a 24 horas de incubación de V. cholerae O1 (cepa SOS-833, CNRB, Costa Rica) y se evaluó la utilidad del análisis microscópico de la motilidad para racionalizar el uso de las técnicas rápidas. Resultados. La sensibilidad, tanto de SMART directo como de PDK directo, fue de 100% en muestras de heces diarreicas y semiformadas y en contenido intestinal de cadáveres. Con estas muestras, el procedimiento SMART directo mostró una especificidad de 100%, mientras que con el PDK directo esta fue de 85,7% a 77,4%, en dependencia del tipo de muestra. Los resultados positivos falsos obtenidos mediante PDK directo resultaron negativos con PDK-6 y PDK-18. Entre los hisopados rectales y fecales de personas con y sin diarrea o que recibieron tratamiento previo con antibióticos se observaron tres resultados negativos falsos con SMART-6 y dos con PDK-6, los cuales resultaron positivos mediante SMART-18 y PDK-18, respectivamente. Ambos sistemas mostraron una concordancia excelente (índice kappa superior a 0,9) en las diferentes modalidades evaluadas. La sensibilidad analítica de ambos sistemas fue de 6 107 ufc/mL de V. cholerae O1, lo que concordó con la observación microscópica de 10 microorganismos o más con motilidad típica de vibriones por campo (aumento de 1000). Las muestras con menos de 10 microorganismos con motilidad típica de vibriones tenían concentraciones entre 6 103 y 6 106 ufc/mL y solo resultaron positivas después de un enriquecimiento de 6­18 horas. Se propone una estrategia para establecer la presencia de Vibrio cholerae O1 en un tiempo inferior al de los métodos convencionales, con valores predictivos positivo y negativo de 100%. Conclusiones. Los sistemas SMART y PDK permiten llegar a un diagnóstico certero de cólera en poco tiempo, no requieren de instrumental complejo ni de personal técnico altamente calificado y funcionan satisfactoriamente en condiciones de campo. Mediante la estrategia propuesta se pueden aumentar la especificidad y la sensibilidad de estos sistemas y se reducen los costos del diagnóstico, lo que permite recomendar su empleo para la vigilancia del cólera en áreas con escasos recursos, donde esta enfermedad constituye un grave problema de salud pública

Objectives. To compare the performance of two rapid systems for the diagnosis of cholera with the culture method, and to propose a strategy for improving the specificity and sensitivity of these systems and reducing the costs involved in making a diagnosis. Methods. The following institutions participated in the study: the National Bacteriology Referral Center (Centro Nacional de Referencia en Bacteriología, CNRB) of the Costa Rican Institute for Research and Teaching in Nutrition and Health (Instituto Costarricense de Investigación y Enseñanza en Nutrición y Salud, INCIENSA) and various hospitals in the provinces of Alajuela, Guanacaste and San José, in Costa Rica. A total of 237 feces samples were used to asses the performance of two tests for the rapid detection of Vibrio cholerae 01: the Pathogen Detection Kit© (PDK, Intelligent Monitoring Systems, Gainesville, Florida, USA) and Cholera-SMART© (New Horizons Diagnostics Corp., Columbia, Maryland, USA), both when applied directly (direct SMART and direct PDK) and when applied to specimens cultured in brothenriched medium for 6 hours (SMART-6 and CPK-6) and for 18 hours (SMART-18 and PDK-18) at 37 °C in alkaline peptone water. Liquid and partially formed stools were cultured and examined by means of the rapid direct test; when the initial result for periods of 6 and 18 hours. Rectal and fecal swabs were obtained from feces cultured in enriched-broth medium for 6 and 18 hours. In addition, we studied the sensitivity of the rapid testing systems by using pure cultures of V. cholerae 01 (strain SOS-833, CNRB, Costa Rica) that were incubated for 18 to 24 hours, and we assessed the usefulness of observing motility under the microscope in order to rationalize the use of rapid methods. Results. The sensitivity of the direct SMART test and of the direct PDK test was 100% when samples obtained from liquid and partially formed stools and from the intestinal contents of dead bodies were used. With these samples, the direct SMART procedure showed a specificity of 100%, whereas the direct PDK procedure showed a specificity that ranged from 85.7% to 77.4%, depending on the type of sample. False positives obtained with the direct PDK method turned out to be negative with PDK-6 and PDK-18. Among the rectal and fecal swabs of persons with and without diarrhea or who had received prior treatment with antibiotics, three results that were negative with the SMART-6 procedure and two that were negative with the PDK-6 procedure turned out to be positive with the SMART-18 and PDK-18 procedures, respectively. Both systems showed excellent concordance (kappa index above 0.9) throughout. Both systems were sensitive to 6 107 colony-forming units per milliliter (cfu/mL), which was concordant with the microscopic observation of 10 microorganisms or more per field with the type of motility that characterizes vibrios (at 1 000 magnification). Samples having fewer than 10 microorganisms with the motility that characterizes vibrios had concentrations between 6 103 and 6 106 cfu/mL and became positive only after incubation in enriched-broth medium for 6 to 18 hours. We propose a strategy for diagnosing the presence of V. cholerae 01 infection in less time than it takes with traditional methods, with positive and negative predictive values of 100%. Conclusions. The SMART and PDK systems make it possible to accurately diagnose cholera quickly, don't require sophisticated equipment or highly qualified technical personnel, and perform satisfactorily in field conditions. Through the proposed strategy, it becomes possible to improve the specificity and sensitivity of these systems and to reduce the cost of making a diagnosis, thus making them suitable for use in cholera surveillance in low-income settings where this disease is a serious public health problem

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Pathogenic strains of Vibrio cholerae O139 possess the cholera toxin A subunit (ctxA) gene as well as the gene for toxin co-regulated pili (tcpA). We report the isolation of a ctxA-negative, tcpA-negative V. cholerae O139 strain (INDREI) from a patient in Mexico diagnosed with gastrointestinal illness. Certain phenotypic characteristics of this strain were identical to those of V. cholerae O1 biotype El Tor. Unlike ctxA-positive V. cholerae O139 strains, this strain was sensitive to a wide panel of antibiotics, including ampicillin, chloramphenicol, ciprofloxacin, gentamicin, furazolidone, nalidixic acid, nitrofurantoin, tetracycline, trimethoprim-sulfamethoxazole, and streptomycin, but was resistant to polymyxin B. Ribotype and pulsed-field gel electrophoresis profiles of INDRE1 differed from those of ctxA-positive V. cholerae O139 and other V. cholerae strains. Phenotypic characteristics of the Mexico strain were similar to those reported for V. cholerae O139 isolates from Argentina and Sri Lanka.

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