Foodborne Viruses and Somatic Coliphages Occurrence in Fresh Produce at Retail from Northern Mexico.

Foodborne disease outbreaks linked to consumption of vegetables have been often attributed to human enteric viruses, such as Norovirus (NoV), Hepatitis A virus (HAV), and Rotavirus (RoV). Information about the occurrence of these viruses is scarce in many fresh-producing countries. Viral contamination detection of indicators, such as somatic coliphages, could indirectly reflect the presence of viral pathogens, being a valuable tool for better viral risk assessment in food industry. This study aimed to establish the occurrence and correlation of foodborne viruses and somatic coliphages in leafy greens in northern Mexico. A total of 320 vegetable samples were collected, resulting in 80 composite rinses, 40 of lettuce and 40 of parsley. Somatic coliphages were determined using the EPA 1602 method, while foodborne viruses (HAV, RoV, NoV GI, and GII) were determined by qPCR. The occurrence of RoV was 22.5% (9/40, mean 2.11 log gc/g) in lettuce and 20% (8/40, mean 1.91 log gc/g) in parsley. NoV and HAV were not detected in any samples. Somatic coliphages were present in all lettuce and parsley samples, with mean levels of 1.85 log PFU/100 ml and 2.28 log PFU/100 ml, respectively. Spearman analysis established the correlation of somatic coliphages and genomic copies of RoV, resulting in an r2 value of - 0.026 in lettuce and 0.349 in parsley. Although NoV or HAV were undetected in the samples, the presence of RoV is a matter of concern as leafy greens are usually eaten raw, which poses a potential risk of infection.

Mathematical model of interaction Escherichia coli and Coliphages.

We propose a mathematical model based in ordinary differential equations between bacterial pathogen and Bacteriophages to describe the infection dynamics of these populations, for which we use a nonlinear function with an inhibitory effect. We study the stability of the model using the Lyapunov theory and the second additive compound matrix and perform a global sensitivity analysis to elucidate the most influential parameters in the model, besides we make a parameter estimation using growth data of Escherichia coli (E.coli) bacteria in presence of Coliphages (bacteriophages that infect E.coli) with different multiplicity of infection. We found a threshold that indicates whether the bacteriophage concentration will coexist with the bacterium (the coexistence equilibrium) or become extinct (phages extinction equilibrium), the first equilibrium is locally asymptotically stable while the other is globally asymptotically stable depending on the magnitude of this threshold. Beside we found that the dynamics of the model is particularly affected by infection rate of bacteria and Half-saturation phages density. Parameter estimation show that all multiplicities of infection are effective in eliminating infected bacteria but the smaller one leaves a higher number of bacteriophages at the end of this elimination.

Early antitermination in the atypical coliphage mEp021 mediated by the Gp17 protein.

The coliphage mEp021 belongs to a phage group with a unique immunity repressor, and its life cycle requires the host factor Nus. mEp021 has been classified as non-lambdoid based on its specific characteristics. The mEp021 genome carries a gene encoding an Nλ-like antiterminator protein, termed Gp17, and three nut sites (nutL, nutR1, and nutR2). Analysis of plasmid constructs containing these nut sites, a transcription terminator, and a GFP reporter gene showed high levels of fluorescence when Gp17 was expressed, but not in its absence. Like lambdoid N proteins, Gp17 has an arginine-rich motif (ARM), and mutations in its arginine codons inhibit its function. In infection assays using the mutant phage mEp021ΔGp17::Kan (where gp17 has been deleted), gene transcripts located downstream of transcription terminators were obtained only when Gp17 was expressed. In contrast to phage lambda, mEp021 virus particle production was partially restored (>1/3 relative to wild type) when nus mutants (nusA1, nusB5, nusC60, and nusE71) were infected with mEp021 and Gp17 was overexpressed. Our results suggest that RNA polymerase reads through the third nut site (nutR2), which is more than 7.9 kbp downstream of nutR1.

Coliphages as viral indicators in municipal wastewater: A comparison between the ISO and the USEPA methods based on a systematic literature review.

The use of traditional faecal indicator bacteria as surrogate organisms for pathogenic viruses in domestic wastewater has been noted as a problematic as concentrations and removal rates of bacteria and viruses do not seem to correlate. In this sense, bacteriophages (phages) emerge as potential viral indicators, as they are commonly found in wastewater in high levels, and can be quantified using simple, fast, low-cost methods. Somatic and F-specific coliphages comprise groups of phages commonly used as indicators of water quality. There are two internationally recognised methods to detect and enumerate coliphages in water samples, the International Standardization Organization (ISO) and the US Environmental Protection Agency (USEPA) methods. Both methods are based on the lysis of specific bacterial host strains infected by phages. Within this context, this systematic literature review aimed at gathering concentrations in raw and treated domestic wastewater (secondary, biological treatment systems and post-treatment systems), and removal efficiencies of somatic and F-specific coliphages obtained by ISO and USEPA methods, and then compare both methods. A total of 33 research papers were considered in this study. Results showed that the ISO method is more commonly applied than the USEPA method. Some discrepancies in terms of concentrations and removal efficiencies were observed between both methods. Higher removal rates were observed for both somatic and F-specific coliphages in activated sludge systems when using the USEPA method compared to the ISO method; in other secondary (biological) treatment systems, this was observed only for F-specific coliphages. The use of different standardised methods available might lead to difficulties in obtaining and comparing phage data in different conditions and locations. Future research comparing both ISO and USEPA methods as well as viral and bacterial pathogens and indicators in WWTP is recommended.

Occurrence of pathogenic microorganisms in small drinking-water systems in Costa Rica.

This study describes the quality of drinking water sampled over 2 years (2018 and 2019) from 20 ASADAS (Spanish acronym for Administrative Associations for Water and Sewer Systems) in Costa Rica. The analysis included Rotavirus (RV), somatic coliphages, fecal coliforms, and Escherichia coli. The ASADAS were categorized into three regions as temperate rainy (region 1), tropical rainy (region 2), and tropical rainy and dry (region 3) according to biogeographic classification. The concentrations of fecal coliforms and E. coli were higher in samples from surface water sources from the ASADAS in region 3 compared to regions 1 and 2. RV-positive samples (24/296) were detected in drinking-water samples from regions 2 and 3 during dry and transition seasons, with higher concentrations more frequently in the dry season. In addition, somatic coliphages were detected in samples from the three regions, with higher concentrations in region 2. Furthermore, a statistically significant relationship was found between somatic coliphages and diarrheal cases, classified as outbreaks or alerts in the region. Thus, the results confirmed that somatic coliphages are a good indicator of the presence of diarrhea cases in a specific region.

Characterization of a Coliphage AS1 isolated from sewage effluent in Pakistan / Caracterização de um colifago AS1 isolado de efluente de esgoto no Paquistão

The emergence of multi-drug resistant (MDR) bacterial strains, which are posing a global health threat has developed the interest of scientists to use bacteriophages instead of conventional antibiotics therapy. In light of an increased interest in the use of phage as a bacterial control agent, the study aimed to isolate and characterize lytic phages from sewage effluent. During the current study, bacteriophage AS1 was isolated from sewage effluent against E.coli S2. The lytic activity of phageAS1 was limited to E.coli S2 strain showing monovalent behavior. The calculated phage titer was 3.5×109 pfu/ml. PhageAS1 was stable at a wide range of pH and temperature. The maximum stability was recorded at 37ºC and pH 7.0, while showing its normal lytic activity at temperature 60ºC and from pH 5.0 to 11.0 respectively. At temperature 70ºC, phage activity was somewhat reduced whereas, further increase in temperature and decrease or increase in pH completely inactivated the phage. From the current study, it was concluded that waste water is a best source for finding bacteriophages against multi-drug resistant bacterial strains and can be used as bacterial control agent.

O surgimento de cepas bacterianas multirresistentes (MDR), que representam uma ameaça global à saúde, desenvolveu o interesse dos cientistas em usar bacteriófagos em vez da terapia convencional com antibióticos. Diante do crescente interesse no uso de fago como agente de controle bacteriano, o estudo visou isolar e caracterizar fagos líticos de efluente de esgoto. Durante o estudo atual, o bacteriófago AS1 foi isolado de efluente de esgoto contra E. coli S2. A atividade lítica de phageAS1 foi limitada à cepa E. coli S2, apresentando comportamento monovalente. O título de fago calculado foi de 3,5 x 109 ufp/ml. PhageAS1 foi estável em uma ampla faixa de pH e temperatura. A estabilidade máxima foi registrada a 37ºC e pH 7,0, enquanto mostrou atividade lítica normal em temperatura de 60ºC e pH 5,0 a 11,0, respectivamente. Na temperatura de 70ºC, a atividade do fago foi um pouco reduzida, enquanto o aumento adicional da temperatura e a diminuição ou aumento do pH inativaram completamente o fago. Com base no estudo atual, concluiu-se que a água residual é a melhor fonte para encontrar bacteriófagos contra cepas bacterianas multirresistentes e pode ser usada como agente de controle bacteriano.

Characterization of a Coliphage AS1 isolated from sewage effluent in Pakistan / Caracterização de um colifago AS1 isolado de efluente de esgoto no Paquistão

The emergence of multi-drug resistant (MDR) bacterial strains, which are posing a global health threat has developed the interest of scientists to use bacteriophages instead of conventional antibiotics therapy. In light of an increased interest in the use of phage as a bacterial control agent, the study aimed to isolate and characterize lytic phages from sewage effluent. During the current study, bacteriophage AS1 was isolated from sewage effluent against E.coli S2. The lytic activity of phageAS1 was limited to E.coli S2 strain showing monovalent behavior. The calculated phage titer was 3.5×109 pfu/ml. PhageAS1 was stable at a wide range of pH and temperature. The maximum stability was recorded at 37ºC and pH 7.0, while showing its normal lytic activity at temperature 60ºC and from pH 5.0 to 11.0 respectively. At temperature 70ºC, phage activity was somewhat reduced whereas, further increase in temperature and decrease or increase in pH completely inactivated the phage. From the current study, it was concluded that waste water is a best source for finding bacteriophages against multi-drug resistant bacterial strains and can be used as bacterial control agent.(AU)

O surgimento de cepas bacterianas multirresistentes (MDR), que representam uma ameaça global à saúde, desenvolveu o interesse dos cientistas em usar bacteriófagos em vez da terapia convencional com antibióticos. Diante do crescente interesse no uso de fago como agente de controle bacteriano, o estudo visou isolar e caracterizar fagos líticos de efluente de esgoto. Durante o estudo atual, o bacteriófago AS1 foi isolado de efluente de esgoto contra E. coli S2. A atividade lítica de phageAS1 foi limitada à cepa E. coli S2, apresentando comportamento monovalente. O título de fago calculado foi de 3,5 x 109 ufp/ml. PhageAS1 foi estável em uma ampla faixa de pH e temperatura. A estabilidade máxima foi registrada a 37ºC e pH 7,0, enquanto mostrou atividade lítica normal em temperatura de 60ºC e pH 5,0 a 11,0, respectivamente. Na temperatura de 70ºC, a atividade do fago foi um pouco reduzida, enquanto o aumento adicional da temperatura e a diminuição ou aumento do pH inativaram completamente o fago. Com base no estudo atual, concluiu-se que a água residual é a melhor fonte para encontrar bacteriófagos contra cepas bacterianas multirresistentes e pode ser usada como agente de controle bacteriano.(AU)

Characterization of a Coliphage AS1 isolated from sewage effluent in Pakistan / Caracterização de um colifago AS1 isolado de efluente de esgoto no Paquistão

The emergence of multi-drug resistant (MDR) bacterial strains, which are posing a global health threat has developed the interest of scientists to use bacteriophages instead of conventional antibiotics therapy. In light of an increased interest in the use of phage as a bacterial control agent, the study aimed to isolate and characterize lytic phages from sewage effluent. During the current study, bacteriophage AS1 was isolated from sewage effluent against E.coli S2. The lytic activity of phageAS1 was limited to E.coli S2 strain showing monovalent behavior. The calculated phage titer was 3.5×109 pfu/ml. PhageAS1 was stable at a wide range of pH and temperature. The maximum stability was recorded at 37ºC and pH 7.0, while showing its normal lytic activity at temperature 60ºC and from pH 5.0 to11.0 respectively. At temperature 70ºC, phage activity was somewhat reduced whereas, further increase in temperature and decrease or increase in pH completely inactivated the phage. From the current study, it was concluded that waste water is a best source for finding bacteriophages against multi-drug resistant bacterial strains and can be used as bacterial control agent.

O surgimento de cepas bacterianas multirresistentes (MDR), que representam uma ameaça global à saúde, desenvolveu o interesse dos cientistas em usar bacteriófagos em vez da terapia convencional com antibióticos. Diante do crescente interesse no uso de fago como agente de controle bacteriano, o estudo visou isolar e caracterizar fagos líticos de efluente de esgoto. Durante o estudo atual, o bacteriófago AS1 foi isolado de efluente de esgoto contra E. coli S2. A atividade lítica de phageAS1 foi limitada à cepa E. coli S2, apresentando comportamento monovalente. O título de fago calculado foi de 3,5 x 109 ufp/ml. PhageAS1 foi estável em uma ampla faixa de pH e temperatura. A estabilidade máxima foi registrada a 37ºC e pH 7,0, enquanto mostrou atividade lítica normal em temperatura de 60ºC e pH 5,0 a 11,0, respectivamente. Na temperatura de 70ºC, a atividade do fago foi um pouco reduzida, enquanto o aumento adicional da temperatura e a diminuição ou aumento do pH inativaram completamente o fago. Com base no estudo atual, concluiu-se que a água residual é a melhor fonte para encontrar bacteriófagos contra cepas bacterianas multirresistentes e pode ser usada como agente de controle bacteriano.

Characterization of a Coliphage AS1 isolated from sewage effluent in Pakistan.

The emergence of multi-drug resistant (MDR) bacterial strains, which are posing a global health threat has developed the interest of scientists to use bacteriophages instead of conventional antibiotics therapy. In light of an increased interest in the use of phage as a bacterial control agent, the study aimed to isolate and characterize lytic phages from sewage effluent. During the current study, bacteriophage AS1 was isolated from sewage effluent against E.coli S2. The lytic activity of phageAS1 was limited to E.coli S2 strain showing monovalent behavior. The calculated phage titer was 3.5×109 pfu/ml. PhageAS1 was stable at a wide range of pH and temperature. The maximum stability was recorded at 37ºC and pH 7.0, while showing its normal lytic activity at temperature 60ºC and from pH 5.0 to11.0 respectively. At temperature 70ºC, phage activity was somewhat reduced whereas, further increase in temperature and decrease or increase in pH completely inactivated the phage. From the current study, it was concluded that waste water is a best source for finding bacteriophages against multi-drug resistant bacterial strains and can be used as bacterial control agent.

A Somatic Coliphage Threshold Approach To Improve the Management of Activated Sludge Wastewater Treatment Plant Effluents in Resource-Limited Regions.

Effective wastewater management is crucial to ensure the safety of water reuse projects and effluent discharge into surface waters. Multiple studies have demonstrated that municipal wastewater treatment with conventional activated sludge processes is inefficient for the removal of a wide spectrum of viruses in sewage. In this study, a well-accepted statistical approach was used to investigate the relationship between viral indicators and human enteric viruses during wastewater treatment in a resource-limited region. Influent and effluent samples from five urban wastewater treatment plants (WWTPs) in Costa Rica were analyzed for somatic coliphage and human enterovirus, hepatitis A virus, norovirus genotypes I and II, and rotavirus. All WWTPs provide primary treatment followed by conventional activated sludge treatment prior to discharge into surface waters that are indirectly used for agricultural irrigation. The results revealed a statistically significant relationship between the detection of at least one of the five human enteric viruses and somatic coliphage. Multiple logistic regression and receiver operating characteristic curve analysis identified a threshold of 3.0 × 103 (3.5 log10) somatic coliphage PFU per 100 ml, which corresponded to an increased likelihood of encountering enteric viruses above the limit of detection (>1.83 × 102 virus targets/100 ml). Additionally, quantitative microbial risk assessment was executed for farmers indirectly reusing WWTP effluent that met the proposed threshold. The resulting estimated median cumulative annual disease burden complied with World Health Organization recommendations. Future studies are needed to validate the proposed threshold for use in Costa Rica and other regions.IMPORTANCE Effective wastewater management is crucial to ensure safe direct and indirect water reuse; nevertheless, few countries have adopted the virus log reduction value management approach established by the World Health Organization. In this study, we investigated an alternative and/or complementary approach to the virus log reduction value framework for the indirect reuse of activated sludge-treated wastewater effluent. Specifically, we employed a well-accepted statistical approach to identify a statistically sound somatic coliphage threshold value which corresponded to an increased likelihood of human enteric virus detection. This study demonstrates an alternative approach to the virus log reduction value framework which can be applied to improve wastewater reuse practices and effluent management.

Recombinant antibodies against Iranian cobra venom as a new emerging therapy by phage display technology

Background:The production of antivenom from immunized animals is an established treatment for snakebites; however, antibody phage display technology may have the capacity to delivery results more quickly and with a better match to local need. Naja oxiana, the Iranian cobra, is a medically important species, responsible for a significant number of deaths annually. This study was designed as proof of principle to determine whether recombinant antibodies with the capacity to neutralize cobra venom could be isolated by phage display.Methods:Toxic fractions from cobra venom were prepared by chromatography and used as targets in phage display to isolate recombinant antibodies from a human scFv library. Candidate antibodies were expressed in E. coli HB2151 and purified by IMAC chromatography. The selected clones were analyzed in in vivo and in vitro experiments.Results:Venom toxicity was contained in two fractions. Around a hundred phage clones were isolated against each fraction, those showing the best promise were G12F3 and G1F4. While all chosen clones showed low but detectable neutralizing effect against Naja oxiana venom, clone G12F3 could inhibit PLA2 activity.Conclusion:Therefore, phage display is believed to have a good potential as an approach to the development of snake antivenom.(AU)

Aptness of Escherichia coli host strain CB390 to detect total coliphages in Colombia.

Fecal bacteria have been used for more than a century as indicators of fecal contamination in water. In recent years, the monitoring of somatic and F-specific coliphages has been gradually included in guidelines and regulations as an additional parameter to reinforce water safety. The Escherichia coli host strain CB390 was tailored to detect both somatic and F-specific coliphages in a single test. The efficacy of this strain for bacteriophage detection, previously evaluated in Western Europe and North America, was assessed here for the first time in South America. The detection of somatic and F-specific coliphages by the strain CB390, as well as by standardized methods, was performed in drinking and river water and municipal and abattoir wastewaters. No statistical difference was found in the numbers of total coliphages detected by strain CB390 and the sum of somatic and F-specific coliphages determined separately by the standardized ISO methods. The data presented here provide further validation of the effectiveness of the host strain E. coli CB390 for the detection of total coliphages in waters in a single test and demonstrate its suitability for application in upper-middle income countries of the Americas (World Bank category).

Somatic Coliphage Profiles of Produce and Environmental Samples from Farms in Northern México.

Somatic coliphages were quantified in 459 produce and environmental samples from 11 farms in Northern Mexico to compare amounts of somatic coliphages among different types of fresh produce and environmental samples across the production steps on farms. Rinsates from cantaloupe melons, jalapeño peppers, tomatoes, and the hands of workers, soil, and water were collected during 2011-2012 at four successive steps on each farm, from the field before harvest through the packing facility, and assayed by FastPhage MPN Quanti-tray method. Cantaloupe farm samples contained more coliphages than jalapeño or tomato (p range <0.01-0.03). Across production steps, jalapeños had higher coliphage percentages before harvest than during packing (p = 0.03), while tomatoes had higher coliphage concentrations at packing than all preceding production steps (p range <0.01-0.02). These findings support the use of targeted produce-specific interventions at multiple points in the process of growing and packing produce to reduce the risk of enteric virus contamination and improve food safety during fruit and vegetable production.

Relación entre la presencia de colifagos en agua para consumo humano, las lluvias y las diarreas agudas en Costa Rica / Relation between coliphages presence in drinking water, rain and acute diarrhea in Costa Rica

ResumenLa calidad del agua de consumo es una de las preocupaciones más grandes a las que se enfrentan los entes de salud. En Costa Rica, a pesar de la buena calidad de las fuentes de agua y del agua de consumo, se siguen registrando numerosos casos de diarrea anualmente. En este estudio se realizó un análisis de indicadores de contaminación virales y bacterianos, en dos comunidades del Valle Central con acueducto propio. Se analizaron en total 24 muestras de agua por cada comunidad, durante un año, 12 en la fuente de agua y 12 en el agua tratada. La frecuencia de diarreas en cada comunidad fue comparada con la calidad microbiológica del agua consumida, encontrándose ausencia de indicadores bacterianos en muestras de agua tratada, mientras que la presencia de indicadores virales en el agua de consumo (con cloro residual) se relaciona con un incremento en el número de casos de diarreas, adicionalmente se analizó el comportamiento de las diarreas de acuerdo a la precipitación promedio con el fin de observar posibles patrones de estacionalidad. Estos hallazgos respaldan la necesidad de ampliar el monitoreo de calidad microbiológica del agua con otro tipo de indicadores de contaminación que correlacionen mejor con otros agentes patógenos de transmisión hídrica, principalmente en época seca, periodo en que se observan más casos de diarrea.

AbstractThe quality of potable water is one of the most important issues of public health. In Costa Rica, in spite of having good sources of water and good quality of water for consumption, there continues to be a significant number of diarrhea cases annually. In this investigation, viral and bacterial analyses of indicators of contamination were conducted in two rural communities with their own aqueducts in the Central Valley of Costa Rica. Twenty-four samples were taken from each community during the year; 12 from the water sources and 12 from the treated waters. The frequency of diarrhea in each community was compared to the microbiological quality of the water being consumed, noting an absence of bacterial indicators in samples of treated water, while the presence of viral indicators in the water (with residual chlorine) were associated with an incremental increase in the number of cases of diarrhea. The analysis also included measurements of average monthly rainfall with the aim of observing possible associations between seasonal conditions and the incidence of diarrhea cases. These findings indicate the need to increase the monitoring of the microbiological quality of the water with other types of indicators of contamination that correlate better with other pathogens transmitted by water, especially in the dry seasons when a greater number of cases of diarrhea are observed.

Metodología rápida y sencilla para la determinación de colifagos somáticos como indicadores de contaminación fecal en una planta de tratamiento de agua localizada al noreste colombiano / Quick and easy methodology to determine somatic coliphages as indicators of fecal contamination in a water treatment plant located in northeastern Colombia

Introducción: En el presente artículo se propone una metodología rápida y sencilla para determinar colifagos sómaticos en las diferentes etapas del tratamiento de agua de una planta del Nordeste Colombiano, con el fin de verificar la calidad del proceso de potabilización. Materiales y métodos: Se empleó la técnica de filtración por membrana para obtener el contenido fágico y posteriormente se cuantificó la presencia de los virus a través de la metodología de capa simple usando agar tripticasa de soya modificado. Resultados: El 88% de las muestras de agua analizadas resultaron ser positivas en alguna de las etapas del proceso de potabilización. La presencia de fagos aumentó en época de lluvias con recuentos máximos de 1310 UFP/100mL en la etapa de captación del agua y de 2,5 UFP/100mL en la etapa de cloración. La prueba no-paramétrica Chi cuadrado de homogeneidad en distribución corroboró estadísticamente la igualdad en distribución de las muestras positivas encontradas en cada una de las etapas de potabilización (p>5%) pues el muestreo fue fijado por indicador. Conclusión: La metodología empleada permitió en tan solo 12 horas determinar la presencia de colifagos somáticos en las diferentes etapas de tratamiento de agua de la planta objeto de estudio. Además, se comprobó la elevada resistencia de estas nanomáquinas naturales a los procesos de desinfección, que mostraron persistencia en etapas de almacenamiento y distribución, especialmente en época de lluvias.

Introduction: In this article a quick and simple method is proposed to determine somatic coliphages in the different stages of water treatment in a plant in the northeast of Colombia, in order to verify the quality of the treatment process. Materials and methods: The technique of membrane filtration was used to get the phage content and subsequently through the methodology of single layer using tryptic soy agar modified, the presence of the virus was quantified. Results: 88% of the water samples analyzed were found to be positive in any of the stages of purification. The presence of phages increased during the rainy season with maximum counts of 1310 UFP / 100 mL at the stage of uptake of water and 2.5 UFP / 100 mL in the chlorination stage. Non-parampetrica Chi square homogeneous distribution test confirmed statistically equal distribution of positive samples found in each of the stages of purification (p> 5%) because the sample was determined by indicator. Conclusion: The methodology allowed to determine the presence of somatic coliphages at different stages of water treatment in the plant under study in only 12 hours. In addition, the high resistance of these natural nanomachines to the processes of disinfection was found, which showed persistence in warehousing and distribution stages, especially in the rainy season.

Prospective epidemiological pilot study on the morbidity of bathers exposed to tropical recreational waters and sand.

A prospective cohort epidemiological pilot study was performed at three tropical beaches with point- and non-point-sources of fecal pollution to characterize the risk of illness among swimmers and non-swimmers. There was an increased risk of illness in swimmers as compared to non-swimmers, even when waters met current microbial standards for recreational water quality. Illnesses included gastrointestinal (GI), skin and respiratory symptoms, earache and fever. Odds ratios (ORs) ranged from 0.32 to 42.35 (GI illness), 0.69 to 3.12 (skin infections), 0.71 to 3.21 (respiratory symptoms), 0.52 to 15.32 (earache) and 0.80 to 1.68 (fever), depending on the beach sampled. The indicators that better predicted the risks of symptoms (respiratory) in tropical recreational waters were total (somatic and male-specific) coliphages (OR = 1.56, p < 0.10, R(2) = 3.79%) and Escherichia coli (OR = 1.38, p < 0.10, R(2) = 1.97%). The present study supports the potential of coliphages as good predictors of risks of respiratory illness in tropical recreational waters. This is the first study that has determined risks of illness after exposure to tropical recreational waters with point- and non-point sources of fecal contamination. The results give an opportunity to perform epidemiological studies in tropical recreational waters in Puerto Rico which can include more participants and other indicators and detection techniques.

Removal of bacteria, protozoa and viruses through a multiple-barrier household water disinfection system.

Municipal water disinfection systems in some areas are not always able to meet water consumer needs, such as ensuring distributed water quality, because household water management can be a contributing factor in water re-contamination. This fact is related to the storage options that are common in places where water is scarce or is distributed over limited time periods. The aim of this study is to assess the removal capacity of a multiple-barrier water disinfection device for protozoa, bacteria, and viruses. Water samples were taken from households in Mexico City and spiked with a known amount of protozoa (Giardia cyst, Cryptosporidium oocyst), bacteria (Escherichia coli), and viruses (rotavirus, adenovirus, F-specific ribonucleic acid (FRNA) coliphage). Each inoculated sample was processed through a multiple-barrier device. The efficiency of the multiple-barrier device to remove E. coli was close to 100%, and more than 87% of Cryptosporidium oocysts and more than 98% of Giardia cysts were removed. Close to 100% of coliphages were removed, 99.6% of the adenovirus was removed, and the rotavirus was almost totally removed. An effect of site by zone was detected; this observation is important because the water characteristics could indicate the efficiency of the multiple-barrier disinfection device.

Coverage of diarrhoea-associated Escherichia coli isolates from different origins with two types of phage cocktails.

Eighty-nine T4-like phages from our phage collection were tested against four collections of childhood diarrhoea-associated Escherichia coli isolates representing different geographical origins (Mexico versus Bangladesh), serotypes (69 O, 27 H serotypes), pathotypes (ETEC, EPEC, EIEC, EAEC, VTEC, Shigella), epidemiological settings (community and hospitalized diarrhoea) and years of isolation. With a cocktail consisting of 3 to 14 T4-like phages, we achieved 54% to 69% coverage against predominantly EPEC isolates from Mexico, 30% to 53% against mostly ETEC isolates from a prospective survey in Bangladesh, 24% to 61% against a mixture of pathotypes isolated from hospitalized children in Bangladesh, and 60% coverage against Shigella isolates. In comparison a commercial Russian phage cocktail containing a complex mixture of many different genera of coliphages showed 19%, 33%, 50% and 90% coverage, respectively, against the four above-mentioned collections. Few O serotype-specific phages and no broad-host range phages were detected in our T4-like phage collection. Interference phenomena between the phage isolates were observed when constituting larger phage cocktails. Since the coverage of a given T4-like phage cocktail differed with geographical area and epidemiological setting, a phage composition adapted to a local situation is needed for phage therapy approaches against E. coli pathogens.

Escherichia coli O104:H4 Pathogenesis: an Enteroaggregative E. coli/Shiga Toxin-Producing E. coli Explosive Cocktail of High Virulence.

A major outbreak caused by Escherichia coli of serotype O104:H4 spread throughout Europe in 2011. This large outbreak was caused by an unusual strain that is most similar to enteroaggregative E. coli (EAEC) of serotype O104:H4. A significant difference, however, is the presence of a prophage encoding the Shiga toxin, which is characteristic of enterohemorrhagic E. coli (EHEC) strains. This combination of genomic features, associating characteristics from both EAEC and EHEC, represents a new pathotype. The 2011 E. coli O104:H4 outbreak of hemorrhagic diarrhea in Germany is an example of the explosive cocktail of high virulence and resistance that can emerge in this species. A total of 46 deaths, 782 cases of hemolytic-uremic syndrome, and 3,128 cases of acute gastroenteritis were attributed to this new clone of EAEC/EHEC. In addition, recent identification in France of similar O104:H4 clones exhibiting the same virulence factors suggests that the EHEC O104:H4 pathogen has become endemically established in Europe after the end of the outbreak. EAEC strains of serotype O104:H4 contain a large set of virulence-associated genes regulated by the AggR transcription factor. They include, among other factors, the pAA plasmid genes encoding the aggregative adherence fimbriae, which anchor the bacterium to the intestinal mucosa (stacked-brick adherence pattern on epithelial cells). Furthermore, sequencing studies showed that horizontal genetic exchange allowed for the emergence of the highly virulent Shiga toxin-producing EAEC O104:H4 strain that caused the German outbreak. This article discusses the role these virulence factors could have in EAEC/EHEC O104:H4 pathogenesis.

Actividad antimicrobiana de Weissella confusa y sus metabolitos frente a Escherichia coli y Klebsiella pneumoniae / Antimicrobial activity of Weisella confusa and its metabolites against Escherichia coli and Klebsiella pneumoniae

Con el fin evaluar el campo de aplicación potencial de una bacteria ácido láctica y de sus metabolitos, se realizó la cinética de la actividad antimicrobiana de W. confusa y de sus metabolitos contra E. coli, y K. pneumoniae, dos patógenos causantes de enfermedades transmitidas por alimentos. La producción de W. confusa se realizó por fermentación discontinua en sustrato comercial MRS. Se realizaron tres fermentaciones durante 6 horas, sin aireación, agitación continúa 33°C y 100 rpm. Cada hora de fermentación se separaron tres sustancias biológicas, W. confusa con sus metabolitos (W+W10b), células de W. confusa libres de metabolitos (W) y metabolito (W10b) y se midió la actividad antimicrobiana contra los patógenos E. coli, y K. pneumoniae. Se encontraron diferencias estadísticas significativas entre tratamientos y tiempo de fermentación. Para E. coli el tratamiento W presentó la mayor actividad antimicrobiana, la cual se obtuvo entre la cuarta y sexta hora de fermentación (2.45 cm de diámetro promedio de inhibición). Para K. pneumoniae, los tratamientos W y W+W10b presentaron actividad antimicrobiana entre la cuarta y quinta hora de fermentación, sin diferencia significativa entre ellos. W. confusa y el metabolito W10b demostraron poseer capacidad antimicrobiana contra E. coli y K. pneumoniae, lo cual sugiere que W. confusa y W10b podrían utilizarse como alternativa de bioconservación o bioprotección de alimentos frescos y procesados, para alimentación humana y animal; y podría convertirse en una alternativa al uso de antibióticos para enfermedades causadas por E. coli y K. pneumoniae.

The kinetic of antimicrobial activity of Weissella confusa and their metabolites against E. coli, and K. pneumoniae, (two pathogens causing foodborne illness) was evaluated, in order to know the possible use in food processing. W. confusa was produced by batch fermentation using MRS commercial substrate. Three fermentations, of 6 hours at 33 °C, without aeration, stirring continuously (100 rpm) were performed. Every hour of fermentation, three biological substances, W. confusa with their metabolites (W + W10b), W. confusa free cells metabolites (W), and metabolite (W10b) were separated, and subsequently the antimicrobial activity against pathogenic E. coli and K. pneumoniae was measured. Statistically significant differences between treatments and fermentation time were found. Treatment (W) against E. coli, showed the greatest antimicrobial activity, it was obtained between the fourth and sixth hours of fermentation (2.45 cm diameter average inhibition). In treatments W and W + W10b against K. pneumoniae, statistically significant differences between them were not found. The antimicrobial activity was shown between the fourth and fifth hour of fermentation. W. confusa and W10b have antimicrobial activity against E. coli and K. pneumoniae, suggesting that W and W10b could be used as an alternative to biopreservation or bioprotection of fresh and processed food for human and animal consumption, and could become an alternative to antibiotics used for diseases caused by E. coli and K. pneumoniae.