Development of a low pollution medium for the cultivation of lactic acid bacteria.

Protein rich culture media are employed in the production of lactic acid bacteria (LAB); however, production costs are high. In this work media formulation and evaluation for LAB production were conducted considering physiological properties of lactic acid bacteria. Consumption efficiency (E), yield production (Y) and specific substrate consumption rate (qS) values as response variables were used. Four culture media were used: (1) Man Rogosa Sharp (MRS); (2) cabbage liquor (MC); (3) a new balanced culture medium (MX); and (4) MX supplemented with cabbage liquor (MXC). The culture media were evaluated using two strains: Lactobacillus acidophilus ATCC 4356 and Lactiplantibacillus plantarum ATCC 10241. The EGLU for L. plantarum was 100 % in the three media and YX/S value was 0.02 ± 0.003 in MRS and MX, while YLAC/S was 0.57 ± 0.03 in MRS and 0.51 ± 0.02 in MX. In MXC, the value obtained for YX/S was 0.07 ± 0.002 while YLAC/S was 0.47 ± 0.04. Specific glucose consumption and lactate formation rates for L. plantarum in MRS and MX media did not show significant differences. These results suggest that MX and MXC can be used for efficient production of the LAB at low cost.

Microbiological assessment of honey in México / Calidad microbiológica de la miel en México

Honey is a product used as a natural sweetener and in several regions of Mexico and other countries it is also used as a therapeutic agent. Microbiological contamination of honey can occur during its extraction and handling. Due to the use and consumption of honey we highlighted here the importance of the assessment of its microbiological quality. One thousand nine hundred twenty samples obtained from 8 honey-producing states from Mexico were analyzed. From these samples, 40.5% (777/1920) did not comply with the NMX-036-NORMEX-2006 specification. Forty five percent (777/1920) of the samples did not comply with the mesophilic aerobic microorganism specification, neither did 17% (327/1920) of the samples with the specification for molds and 18.1% (348/1920) with the specification for yeasts. With regard to coliform bacteria, the samples contained less than 3 NMP/g. Two percent of the samples contained lactic acid bacteria (LAB), Clostridium perfringens was observed in amounts of more than 100 CFU/g. None of the samples from the different states contained more than 100 CFU/g of Staphylococcus aureus; Salmonella spp. was absent in all samples. It is important to avoid contamination sources and implement good hygienic practices in order to maintain and improve the quality of Mexican honeys since a large percentage of them are intended for export. If these honeys are intended for therapeutic use, it is necessary to ensure that they comply with all quality parameters and to apply specific treatments that guarantee the removal of any pathogen that may represent a risk to the patients's health.

La miel es un producto que se usa como edulcorante natural y en algunas regiones de México y otros países, como agente terapéutico. La contaminación de la miel con microorganismos se puede presentar durante la extracción y el manejo. Dado el uso y consumo de la miel, se consideró importante evaluar su calidad microbiológica. Se trabajó con 1.920 muestras de miel obtenidas de 8 estados productores en México. De las muestras de miel analizadas, el 40,5% (777/1.920) estuvo fuera de las especificaciones de la NMX-036-NORMEX-2006. Todas esas muestras no cumplían con las especificaciones referidas a mesofílicos aerobios, en tanto que el 17% (327/1920) no cumplían las relativas a mohos y el 18,1% (348/1920), las fijadas para levaduras. En lo que se refiere a los organismos coliformes, las muestras contenían < 3 NMP/g. Se encontró que el 2% contenía BAL; Clostridium perfringens se puso en evidencia en el 12%, con más de 100 UFC/g. En el caso de Staphylococcus aureus, ninguna muestra estuvo por arriba de 100 UFC/g. Salmonella spp. no se detectó en ninguna de las muestras analizadas. Es importante evitar las fuentes de contaminación y observar buenas prácticas de higiene para mantener y mejorar la calidad de las mieles mexicanas, ya que un buen porcentaje de ellas están destinadas a la exportación. Si estas van a ser destinadas a un uso terapéutico es necesario asegurarse de cumplir con los parámetros de calidad y dar tratamientos específicos que aseguren la eliminación de algún patógeno que pueda poner en riesgo la salud del paciente que utilice la miel.

Microbiological assessment of honey in México.

Honey is a product used as a natural sweetener and in several regions of Mexico and other countries it is also used as a therapeutic agent. Microbiological contamination of honey can occur during its extraction and handling. Due to the use and consumption of honey we highlighted here the importance of the assessment of its microbiological quality. One thousand nine hundred twenty samples obtained from 8 honey-producing states from Mexico were analyzed. From these samples, 40.5% (777/1920) did not comply with the NMX-036-NORMEX-2006 specification. Forty five percent (777/1920) of the samples did not comply with the mesophilic aerobic microorganism specification, neither did 17% (327/1920) of the samples with the specification for molds and 18.1% (348/1920) with the specification for yeasts. With regard to coliform bacteria, the samples contained less than 3 NMP/g. Two percent of the samples contained lactic acid bacteria (LAB), Clostridium perfringens was observed in amounts of more than 100CFU/g. None of the samples from the different states contained more than 100CFU/g of Staphylococcus aureus; Salmonella spp. was absent in all samples. It is important to avoid contamination sources and implement good hygienic practices in order to maintain and improve the quality of Mexican honeys since a large percentage of them are intended for export. If these honeys are intended for therapeutic use, it is necessary to ensure that they comply with all quality parameters and to apply specific treatments that guarantee the removal of any pathogen that may represent a risk to the patients's health.

Frecuencia, susceptibilidad antimicrobiana y patrón de adherencia de Salmonella enterica aislada de carne de pollo, res y cerdo de la Ciudad de México / Frequency, antimicrobial susceptibility and adherence patterns of Salmonella enterica isolated from chicken meat, beef and pork from Mexico City

Resumen Introducción: Los alimentos de origen animal frecuentemente están implicados en brotes de salmonelosis. Objetivo: Evaluar la frecuencia de Salmonella enterica en carnes molidas de pollo, res y cerdo (un total de 2.592 muestras) obtenidas de mercados sobre ruedas y supermercados de la Delegación Iztapalapa en la Ciudad de México, determinar la susceptibilidad antimicrobiana y efectuar ensayos de adherencia en las cepas aisladas. Métodos: El aislamiento de S. enterica se hizo de acuerdo a la BAM-FDA, la susceptibilidad antimicrobiana de acuerdo con CLSI y el ensayo de adherencia en células HEp-2 conforme a Baffone y cols., 2001. Resultados: Salmonella enterica fue aislada en 511 del total de muestras analizadas (19,7%), de las cuales 244 (47,7%), 152 (29,7%) y 115 (22,5%) correspondieron a carne molida de pollo, res y cerdo, respectivamente. La mayor frecuencia de resistencia de S. enterica a antimicrobianos fue a ampicilina y cloranfenicol en pollo, perfloxacina y ampicilina en res y carbenicilina, ampicilina, cloranfenicol, cefotaxima y perfloxacina en cerdo. Noventa por ciento de las cepas mostraron un patrón de adherencia agregativo. Conclusión: La frecuencia de S. enterica en productos cárnicos es alta, por lo que es importante la adecuada cocción de la carne para disminuir el riesgo de una salmonelosis.

Background: Food of animal origin is often involved in salmonellosis outbreaks. Aim: To evaluate the frequency of Salmonella enterica in chicken, beef and pork ground meat (a total of 2,592 samples) obtained from travelling markets and supermarkets at the Iztapalapa area of Mexico City, in order to determine the antimicrobial susceptibility and adherence capacity of isolated strains. Methods: Isolation of S. enterica was carried out according to the BAM-FDA, the microbial susceptibility according with CLSI and adherence assay on HEp-2 cell line according with Baffone et al., 2001. Results: S. enterica was isolated from 511 of all the analyzed samples (19.7%), from which 244 (47.7%), 152 (29.7%) and 115 (22.5%) corresponded to chicken, beef and pork ground meat, respectively. The highest frequency of resistance of S. enterica to antimicrobials was to ampicillin and chloramphenicol in chicken, perfloxacin and ampicillin in beef and carbenicillin, ampicillin, chloramphenicol, cefotaxime and perfloxacin in pork. Ninety percent of the strains showed an aggregative adherence pattern on HEp-2 cells. Conclusion: The frequency of S. enterica on meat products is high, which is the reason why a proper cooking of these ground meats is important in order to reduce the risk of acquiring salmonellosis.

[Frequency, antimicrobial susceptibility and adherence patterns of Salmonella enterica isolated from chicken meat, beef and pork from Mexico City]. / Frecuencia, susceptibilidad antimicrobiana y patrón de adherencia de Salmonella enterica aislada de carne de pollo, res y cerdo de la Ciudad de México.

BACKGROUND: Food of animal origin is often involved in salmonellosis outbreaks. AIM: To evaluate the frequency of Salmonella enterica in chicken, beef and pork ground meat (a total of 2,592 samples) obtained from travelling markets and supermarkets at the Iztapalapa area of Mexico City, in order to determine the antimicrobial susceptibility and adherence capacity of isolated strains. METHODS: Isolation of S. enterica was carried out according to the BAM-FDA, the microbial susceptibility according with CLSI and adherence assay on HEp-2 cell line according with Baffone et al., 2001. RESULTS: S. enterica was isolated from 511 of all the analyzed samples (19.7%), from which 244 (47.7%), 152 (29.7%) and 115 (22.5%) corresponded to chicken, beef and pork ground meat, respectively. The highest frequency of resistance of S. enterica to antimicrobials was to ampicillin and chloramphenicol in chicken, perfloxacin and ampicillin in beef and carbenicillin, ampicillin, chloramphenicol, cefotaxime and perfloxacin in pork. Ninety percent of the strains showed an aggregative adherence pattern on HEp-2 cells. CONCLUSION: The frequency of S. enterica on meat products is high, which is the reason why a proper cooking of these ground meats is important in order to reduce the risk of acquiring salmonellosis.

Virulence factors and antimicrobial resistance in environmental strains of Vibrio alginolyticus

Vibrio alginolyticus has acquired increasing importance because this microorganism may be pathogenic to aquatic animals and humans. It has been reported that some V. alginolyticus strains carry virulence genes derived from pathogenic V. cholerae and V. parahaemolyticus strains. In this work V. alginolyticus was isolated from oyster samples acquired from a food-market in Mexico City. Thirty isolates were identified as V. alginolitycus. Strains showed β-haemolysis and proteolytic activity and produced a capsule. Strains displayed swimming and swarming motility and 93.3% of them produced siderophores. Several genes encoding virulence factors were detected using PCR amplification. These included proA, wza, vopD, vopB, hcp, vasH and vgrG genes, which were present in all strains. Other genes had a variable representation: tdh (86.6%), lafA (96.6%), pvsA (62%) and pvuA (16%). The trh gene could not be amplified from any of the strains. The antimicrobial resistance profile revealed that more than 90% of the strains were resistant to beta-lactams antibiotics, 60% to cephalotin, 45% to amikacin, 16% to cephotaxime, and 10% to pefloxacin, while 100% were susceptible to ceftriaxone. The V. alginolyticus strains isolated from oysters showed multiple resistance to antibiotics and several virulence factors described in well-characterized pathogenic vibrios (AU)

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Identification of Enteric Viruses in Foods from Mexico City.

Foodborne viruses are a common and, probably, the most under-recognized cause of outbreaks of gastroenteritis. Among the main foods involved in the transmission of human enteric viruses are mollusks, and fruits and vegetables irrigated with wastewater and/or washed with non-potable water or contaminated by contact with surfaces or hands of the infected personnel during its preparation. In this study, 134 food samples were analyzed for the detection of Norovirus, Rotavirus, and Hepatitis A virus (HAV) by amplification of conserved regions of these viruses. From the 134 analyzed samples, 14 were positive for HAV, 6 for Norovirus, and 11 for Rotavirus. This is the first report in Mexico where emphasis is given to the presence of HAV and Norovirus on perishable foods and food from fisheries, as well as Rotavirus on frozen vegetables, confirming the role of vegetables and bivalve mollusks as transmitting vehicles of enteric viruses.

Virulence factors and antimicrobial resistance in environmental strains of Vibrio alginolyticus.

Vibrio alginolyticus has acquired increasing importance because this microorganism may be pathogenic to aquatic animals and humans. It has been reported that some V. alginolyticus strains carry virulence genes derived from pathogenic V. cholerae and V. parahaemolyticus strains. In this work V. alginolyticus was isolated from oyster samples acquired from a food-market in Mexico City. Thirty isolates were identified as V. alginolitycus. Strains showed ß-haemolysis and proteolytic activity and produced a capsule. Strains displayed swimming and swarming motility and 93.3% of them produced siderophores. Several genes encoding virulence factors were detected using PCR amplification. These included proA, wza, vopD, vopB, hcp, vasH and vgrG genes, which were present in all strains. Other genes had a variable representation: tdh (86.6%), lafA (96.6%), pvsA (62%) and pvuA (16%). The trh gene could not be amplified from any of the strains. The antimicrobial resistance profile revealed that more than 90% of the strains were resistant to beta-lactams antibiotics, 60% to cephalotin, 45% to amikacin, 16% to cephotaxime, and 10% to pefloxacin, while 100% were susceptible to ceftriaxone. The V. alginolyticus strains isolated from oysters showed multiple resistance to antibiotics and several virulence factors described in well-characterized pathogenic vibrios. [Int Microbiol 19(4):191-198 (2016)].

Detection of toxigenic Bacillus cereus strains isolated from vegetables in Mexico City.

Bacillus cereus can cause diarrhea and emetic syndromes after ingestion of food contaminated with it. This ability is due to the production of enterotoxins by this microorganism, these being the hemolysin BL complex, which is involved in the diarrheal syndrome, and cereulide, which is responsible for the emetic syndrome. The detection of genes associated with the production of these toxins can predict the virulence of strains isolated from contaminated food. In this paper, we analyzed 100 samples of vegetables, 25 of each kind (broccoli, coriander, carrot, and lettuce) obtained from different markets in Mexico City and its metropolitan area. B. cereus was isolated in 32, 44, 84, and 68% of the samples of broccoli, carrot, lettuce, and coriander, respectively. The hblA gene (encoding one of the three subunits of hemolysin BL) was amplified in 100% of the B. cereus isolates, and the ces gene (encoding the cereulide) could not be amplified from any of them. This is the first report of B. cereus isolation from the vegetables analyzed in this work and, also, the first report in Mexico of the isolation from vegetables of strains with potential virulence. The results should serve as evidence of the potential risk of consuming these foods without proper treatment.

Rotavirus G2P[4] detection in fresh vegetables and oysters in Mexico City.

Rotaviruses are the principal cause of dehydration caused by diarrhea in children younger than 2 years of age. Although these viral infections have mainly been associated with ingestion of fecally contaminated food and water, few studies have addressed the presence of the virus in food that is consumed raw or slightly cooked. In this work, 30 oyster samples and 33 vegetable samples were examined for the presence of rotavirus genotypes to evaluate their potential to produce gastrointestinal infections. The rotaviruses were identified by reverse transcriptase PCR amplification of the VP7 gene. G and P genotyping was also performed by reverse transcriptase PCR, with a detection sensitivity of up to 15 PFU/ml. Rotaviruses were found in 17 (26.9%) of 63 samples (10 oysters and 7 vegetables). The G2 genotype was found in 11 (64.7%) of 17 of the rotavirus strains, and 16 (94.1%) of 17 had the P[4] genotype. The combined genotypes found most frequently were G2P[4] (10 [58.82%] of 17), GNTP[4] (6 [35.29%] of 17), and G2P[NT] (1 [5.8%] of 17).

Detection of Clostridium perfringens in yearling lamb meat (barbacoa), head, and gut tacos from public markets in Mexico City.

No reports on the incidence of Clostridium perfringens in popularly-consumed food from Mexico City have been published; neither are there any reports that have analyzed food consumed in popular markets and less established restaurants. Therefore, this study is aimed at providing data to evaluate the relevance of C. perfringens as an etiologic agent of food-borne diseases. Of the 650 analyzed samples, 106 (16.3%) were positive for C. perfringens; 6.4% (16/250) isolates were from barbacoa, 19% (38/200) from head, and 13% (52/200) from gut tacos. The presence of C. perfringens in these popular-consumed foods demonstrates its relevance as an etiologic agent of food-borne diseases, and confirms the great sanitary risk involved in their consumption. These results may serve as a basis for the Mexican sanitary authorities to control the microbiological quality of street-made foods.