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Resumen La resistencia a los antimicrobianos (RAM) es un problema de salud pública cada vez más complejo y se considera como una de las mayores amenazas en todo el mundo. El desarrollo de la RA en los patógenos bacterianos es una consecuencia esperada de la adaptación evolutiva, debido a la presencia de este tipo de contaminantes, los antibióticos, en variedad de nichos ecológicos. Además, hay múltiples factores asociados con su origen y diseminación, entre ellos, el uso desmedido y poco regulado de los antibióticos en la medicina humana y veterinaria, así como en la agricultura, la ganadería y la industria. De hecho, recientemente se ha indicado el papel del ambiente como reservorio para genes de RA y bacterias resistentes a antibióticos. En este sentido, el enfoque para contener y controlar este problema tan complejo involucra de forma necesaria a diversas áreas como la medicina, la veterinaria, las ciencias ambientales y sectores de la industria y la economía. En este artículo, se realiza una descripción tanto del problema de la RA y sus elementos causales, como del enfoque multidisciplinario que ha sido propuesto para su manejo en el ámbito global. Se detalla también cómo la RA afecta el desarrollo humano sostenible conforme a la Agenda 2030 formulada por la ONU, en el cumplimiento de algunos de los objetivos del desarrollo sostenible (ODS). Abstract Introduction. Antibiotic resistance (AR) is an increasingly complex public health problem, considered one of the greatest threats globally. The development of AR in bacterial pathogens is an expected consequence of evolutionary adaptation due to antimicrobials contamination in the environment. However, multiple factors are associated with the emergence and dissemination of AR, including excessive and poorly regulated use of antibiotics in human and animal medicine, agriculture, livestock, and industry, among other fields. The role of the environment as a reservoir for the generation and dissemination of AR genes and AR bacteria has recently been indicated. The approach to contain and control this problem involves multiple disciplines such as human and animal medicine, the environment, the industry, and the economy. This article describes the AR problem, the factors associated with its origin, and the multidisciplinary approach proposed for its management at a global level. Also, it will be described how AR affects sustainable development according to the UN 2030 Agenda, in compliance with some of the sustainable development goals.

Resumen (Introducción): El agua para consumo es un determinante de la salud humana; particularmente cuando se obtiene de fuentes superficiales puede ser afectada por contaminantes microbiológicos causantes de enfermedades, los cuales a su vez son influenciados por las condiciones ambientales; sin embargo, estos efectos han sido poco estudiados en Costa Rica. (Objetivo): Determinar la influencia de variables ambientales en las concentraciones de microorganismos patógenos e indicadores fecales en un sistema de abastecimiento de agua para consumo humano. (Metodología): Se realizó un muestreo dos veces al mes durante un año. Se midió la concentración de Cryptosporidium sp., Giardia sp. colifagos somáticos y Escherichia coli. Se analizaron las condiciones espaciales mediante el uso de sistemas de información geográfica, y las condiciones climáticas de manera descriptivas. Para el análisis de datos, se utilizaron técnicas para datos censurados y un modelo de regresión de Poisson para evaluar la influencia de la intensidad de precipitación sobre las concentraciones de microorganismos. (Resultados): Se encuentran diferencias estacionales para colifagos somáticos y E. coli. Cryptosporidium sp. y Giardia sp. son más frecuentes cuando la intensidad de precipitación es débil, mientras que colifagos somáticos y E. coli presentan mayores concentraciones conforme aumenta la intensidad de precipitación. (Conclusiones): Las cuatro microcuencas presentan vulnerabilidad por contaminación con respecto a sus condiciones espaciales y se determina que tanto la estacionalidad climática y la intensidad de la precipitación 24 h antes afectan de manera diferente a cada uno de los microorganismos en estudio, por lo cual, se recomienda su evaluación periódica de forma independientemente. Abstract (Introduction): Drinking water is a determining factor in human health; when obtained from surface sources, it can be affected by microbiological contaminants that cause disease, which in turn could be influenced by environmental conditions. However, these effects have been little studied in Costa Rica. (Objective): To determinate the environmental variability influence in the concentrations of pathogen microorganisms and fecal indicators in a drinking water system. (Methodology): samples were collected twice a month for one year. Cryptosporidium sp., Giardia sp., somatic coliphages, and Escherichia coli concentrations were measure. Spatial conditions were analyzed using geographic information systems, and climate conditions analysis were performed using descriptive technics. To data analysis were used censored data technics and Poisson regression models to evaluate the influence of the rainfall on microorganisms’ concentrations. (Results): seasonal differences were found for somatic coliphages and E. coli. Cryptosporidium sp. and Giardia sp. concentrations were more frequent with weak precipitation intensity. Meanwhile, coliphages and E. coli concentrations were higher when precipitation intensity increase. (Conclusions): The four micro-basins presented pollution vulnerability for their spatial conditions. Besides, study microorganisms were affected differently by climatic seasonality and 24 h previous intensity of the precipitation. Therefore, it is recommended an individual evaluation of them periodically.

Resumen Introducción: la enfermedad diarreica es una de las principales causas de muerte en niños menores de cinco años. Uno de los agentes etiológicos bacterianos más reconocido es Escherichia coli, que puede causar infecciones a través de diversos mecanismos asociados a la adquisición de genes de virulencia. Metodología: se recolectaron muestras de aguas residuales en los afluentes y los efluentes de cinco plantas de tratamiento de la Gran Área Metropolitana, durante tres días consecutivos, en los meses de marzo, mayo, octubre y diciembre del año 2013, a fin de cuantificar la concentración de E. coli, mediante la técnica de fermentación en tubos múltiples, y evaluar la presencia de los genes de virulencia aatA, bfpA, eaeA, ipaH, lt, st, stx1 y stx2, vinculados con patotipos de E. coli diarrogénica, con la técnica de PCR punto final. Resultados: el estudio reveló niveles de E. coli similares a los encontrados en afluentes de otras regiones del mundo, en cambio, los efluentes presentaron concentraciones superiores a los 4 Log10 NMP/100 mL. La circulación anual de genes de virulencia fue relacionada con los patotipos enterotoxigénica, productores de toxina Shiga/enterohemorrágica, enteroagregativa y enteropatógena. No se detectaron genes referentes a la variedad enteroinvasora. Además, se determinó que no todas las plantas de tratamiento remueven significativamente la carga de E. coli. Conclusiones: el flujo de E. coli diarrogénica es común en nuestro país y su diseminación en el ambiente es posible cuando los efluentes se descargan en cuerpos de aguas superficiales Abstract Introduction. Diarrheic disease is one of the most important diseases causes in children under five years. One of the most recognized etiological agents is Escherichia coli. This bacterium has the capacity of causing infections by multiple mechanisms associated with the acquisition of different virulence genes. Methodology: water samples were collected from the affluents and effluents of five wastewater treatment plants located in the Costa Rican Great Metropolitan Area, during three sequential days, in March, May, October, and December from 2013. The aim was to quantify E. colis concentration by multiple tubes fermentation method and to evaluate the presence of aatA, bfpA, eaeA, ipaH, lt, st, stx1, and stx2 virulence genes, related with diarrheagenic E. coli, by PCR end-point method. Results: The study revealed that the levels of E. coli into the affluents are similar to those reported in other world regions. The effluents concentration presented higher than 4 Log10 NMP/100 mL level of E. coli. A year-round circulation of virulence genes was demonstrated, and it was associated with enterotoxigenic, Shiga-toxin producers/ enterohemorrhagic, enteroaggregative, and enteropathogenic pathotypes. Genes related to enteroinvasive variety did not were detected. Not all wastewater treatment plants removed E. coli efficiently. Conclusions: Diarrheagenic E. coli circulation is common in our county with the possibility of dissemination to the environment by effluents wastewater discharge in surface water bodies.

Acute gastrointestinal illness (AGI) continues to be an important cause of morbidity and mortality among all ages. This study applied the principles ofwastewater-based epidemiology for the preventive identification of potential outbreaks of acute viral gastroenteritis and hepatitis A by analyzing the presence of human enteric viruses in influents of small municipal wastewater treatment plants (WWTPs) handling domestic sewage, together with public health reports of acute diarrheal and hepatitis A disease in Costa Rica during 2013. Raw wastewater samples were collected during four seasonal periods with different rainfall levels. The presence of five human enteric viruses (rotavirus A, norovirus GI, norovirus GII, enterovirus, and hepatitis A virus) was studied by endpoint and real-time polymerase chain reaction in influents of five WWTPs. Cases of AGI were analyzed using historical public health reports of endemic levels and quartile ranges for each administrative and territorial area where theWWTPs are located and for its surrounding counties. A tendency for a higher rate of positive viral tests was present 1 week before an increase of AGI cases. Epidemiological weeks categorized asOutbreak (above the 75th percentile) and Success (below the 25th percentile) showed statistically significant differences in terms of positive viral test rates (Wilcoxon test, P50.05). Virologicalmonitoring ofwastewater in smallWWTPs is an appropriate model for epidemiological surveillance of diarrheal and hepatitis A diseases in low- and middle-resource countries. Universidad de Costa Rica/[]/UCR/Costa Rica Instituto Costarricense de Acueductos y Alcantarillados/[]/AyA/Costa Rica UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA)

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.

We report the draft genome sequence of the multidrug-resistant Escherichia coli strain PTA A1517-5, isolated from a wastewater treatment plant in Costa Rica. The genome consists of 4,927,375 bp with a GC content of 50.57% and a total of 4,853 genes. This strain harbors blaCTX-M-115, blaCMY-2, aminoglycoside, tetracycline, and sulfonamide resistance genes.

Two ether and one ester derivatives of the 4-nitro-3-hydroxybenzoic acid were synthesized and characterized. The in vitro antimicrobial and cytotoxic activities of the three novel compounds were also evaluated. The aromatic derivatives showed antibacterial activity against one of the four microorganisms tested and two compounds (C8 and NOBA) had a lower IC50 in HeLa cells. Ministerio de Ciencia y Tecnología/[MAT2013-45657-P]/MICITT/Costa Rica UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA) UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Química UCR::Vicerrectoría de Docencia::Salud::Facultad de Medicina::Escuela de Medicina

Giardia cysts stained with hot carbolfuchsin were used as internal controls in a concentration method for surface water samples. The morphological integrity of stained cysts and the stain's stability and intensity were tested with each of the chemical reagents used in the aluminum sulfate flocculation method. No alterations in morphology or color were noted. The stained cyst preparation has a low cost, high stability, and suitability for both light and immunofluorescent microscopy, making it affordable to researchers in low- and middle-income countries. UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA)

This study used the Canadian Water Quality Index (CWI) to characterize water sampled at three points within the Purires River micro basin, Costa Rica. The first sampling point is located in a high zone with domestic agricultural activities, the second point around the mid-point of the flow of the river, and the third point at the lowest zone with extensive agricultural activities mainly centered on the production of fresh vegetables. Eleven physicochemical parameters (As, Cd, Cr, biochemical oxygen demand (BOD), chemical oxygen demand (COD), NH4+, NO3−, Pb, pH, percent saturation of dissolved oxygen (PSO), and total suspended solids (TSS)) and two microbiological parameters (fecal coliforms and enterococci) were evaluated. We evaluated three different Canadian Water Quality Indexes (CWIs): CWI-1 included only physicochemical parameters, CWI-2 included CWI-1 parameters plus fecal coliforms, and CWI-3 included CWI-2 in addition to enterococci. Statistical analysis of individual parameters showed significant differences between sampling sites. CWI-1 was unable to discriminate between the three sampling points, and characterized the water quality as ‘fair’. CWI-2 was only able to discriminate when the water contained high levels of chemical and microbiological contaminants, while CWI-3 adequately discriminated water quality at each of the sampling points. The evaluation of enterococci together with more traditional water quality parameters enabled better categorization of surface water quality. UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA) UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro en Investigación en Contaminación Ambiental (CICA)

HIV/AIDS transmission among drug users is associated with education, sex practices and substance use. This study examined 159 drugs users’ knowledge, beliefs and sex behavior related to HIV/AIDS risk in Costa Rica. Results showed considerable use of marihuana, alcohol, crack and cocaine and a very low lifetime incidence of other drugs. All substance use patterns were higher than national averages. Respondents showed a high level of knowledge about HIV/AIDS transmission and prevention. However, there did not seem to be a relationship between knowledge and practice. Unprotected sex was common and having an HIV/AIDS test was not a regular practice. Knowledge about HIV/AIDS is not a determinant factor for condom use among this group. It is concluded that having the proper knowledge about transmission and prevention does not guarantee safe sex practices. Further research and public health evidence based practices for HIV/AIDS prevention should target drug user population

Los métodos de recuento en placa capa doble y capa simple de agar, para la cuantificación de colifagos so-máticos en aguas, fueron implementados utilizando como base metodologías estándar. Diferentes variables fueron ensayadas, lo cual permitió la precisión en algunos pasos no incluidos en metodologías estándares. De los hallazgos de mayor importancia, se exponen las consecuencias de utilizar un cultivo de Escherichia coliexcesivamente concentrado y se describe la obtención de un cultivo en fase logarítmica en solo 4 horas de incubación, ajustando la concentración a una densidad óptica de 0,3 a 600nm (3,1 x 10 8 UFC/ mL), o a un McFarland 1 (3,0 x108 UFC/ mL). Se determinó que los controles de colifagos deben ser almacenados a -70 °C para reducir su degradación y que se deben evitar cantidades superiores a 20 mL de mezcla de reacción por plato de Petri, para reducir las burbujas que pueden interferir con la lectura de unidades formadoras de placas (UFP). Se demostró que los colifagos de las muestras de agua pueden almacenarse 48 horas a 4 °C sin que sufran degradación y que en las muestras con altas concentraciones de colifagos no se observa UFP porque se da una lisis confluente de la capa bacteriana. No se encontraron diferencias significativas en la recuperación de colifagos al utilizar un método u otro, pero dichos métodos deben ser evaluados por medio de controles, antes de aplicarlos directamente en el análisis de muestras de agua.Two plate count methods, double layer and single layer of agar for quantification of somatic coliphages in water, were implemented using standard methodologies. Several variables were tested and provided valuable information that was not included in standard methodologies. The most important findings are described, such as the effect of using an excessively concentrated culture of E. coliand production of a log phase culture in only 4 hours of incubation, adjusting the concentration to an optical density of 0.3 at 600 nm (3.1 x 10 8 CFU / mL), or to McFarland 1 (3.0 x 108 CFU / mL). It was determined that coliphages controls must be stored at -70 °C to reduce its degradation. Quantities of reaction mixture exceeding 20 mL per Petri dish must be avoided to prevent interfere with bubbles during the counting of plate forming units (PFU). It was demonstrated that coliphages isolated from water samples can be stored for 48 hours at 4 °C without any degradation, and PFU are not observed in samples with high concentrations of coliphages, because a confluent lysis of the bacte-rial layer. There was no significant difference in the recovery of coliphages using doble layer or single layer methods, but such methods should be evaluated by means of controls, before applying them directly in the analysis of water samples.

artículo (arbitrado)--Universidad de Costa Rica.Instituto de Investigaciones en Salud, 2010 A Multiplex Polymerase Chain Reaction (PCR) assay to be used as an alternative to the conventional culture method in detecting Shigella and enteroinvasive Escherichia coli (EIEC) virulence genes ipaH and ial in lettuce was developed. Efficacy and rapidity of the molecular method were determined as compared to the conventional culture. Lettuce samples were inoculated with different Shigella flexneri concentrations (from 10 CFU/ml to 107 CFU/ml). DNA was extracted directly from lettuce after inoculation (direct-PCR) and after an enrichment step (enrichment PCR). Multiplex PCR detection limit was 104 CFU/ml, diagnostic sensitivity and specificity were 100% accurate. An internal amplification control (IAC) of 100 bp was used in order to avoid false negative results. This method produced results in 1 to 2 days while the conventional culture method required 5 to 6 days. Also, the culture method detection limit was 106 CFU/ml, diagnostic sensitivity was 53% and diagnostic specificity was 100%. In this study a Multiplex PCR method for detection of virulence genes in Shigella and EIEC was shown to be effective in terms of diagnostic sensitivity, detection limit and amount of time as compared to Shigella conventional culture. Universidad de Costa Rica UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA)

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.

Los métodos de recuento en placa capa doble y capa simple de agar, para la cuantificación de colifagos so-máticos en aguas, fueron implementados utilizando como base metodologías estándar. Diferentes variables fueron ensayadas, lo cual permitió la precisión en algunos pasos no incluidos en metodologías estándares. De los hallazgos de mayor importancia, se exponen las consecuencias de utilizar un cultivo de Escherichia coliexcesivamente concentrado y se describe la obtención de un cultivo en fase logarítmica en solo 4 horas de incubación, ajustando la concentración a una densidad óptica de 0,3 a 600nm (3,1 x 10 8 UFC/ mL), o a un McFarland 1 (3,0 x108 UFC/ mL). Se determinó que los controles de colifagos deben ser almacenados a -70 °C para reducir su degradación y que se deben evitar cantidades superiores a 20 mL de mezcla de reacción por plato de Petri, para reducir las burbujas que pueden interferir con la lectura de unidades formadoras de placas (UFP). Se demostró que los colifagos de las muestras de agua pueden almacenarse 48 horas a 4 °C sin que sufran degradación y que en las muestras con altas concentraciones de colifagos no se observa UFP porque se da una lisis confluente de la capa bacteriana. No se encontraron diferencias significativas en la recuperación de colifagos al utilizar un método u otro, pero dichos métodos deben ser evaluados por medio de controles, antes de aplicarlos directamente en el análisis de muestras de agua., Two plate count methods, double layer and single layer of agar for quantification of somatic coliphages in water, were implemented using standard methodologies. Several variables were tested and provided valuable information that was not included in standard methodologies. The most important findings are described, such as the effect of using an excessively concentrated culture of E. coliand production of a log phase culture in only 4 hours of incubation, adjusting the concentration to an optical density of 0.3 at 600 nm (3.1 x 10 8 CFU / mL), or to McFarland 1 (3.0 x 108 CFU / mL). It was determined that coliphages controls must be stored at -70 °C to reduce its degradation. Quantities of reaction mixture exceeding 20 mL per Petri dish must be avoided to prevent interfere with bubbles during the counting of plate forming units (PFU). It was demonstrated that coliphages isolated from water samples can be stored for 48 hours at 4 °C without any degradation, and PFU are not observed in samples with high concentrations of coliphages, because a confluent lysis of the bacte-rial layer. There was no significant difference in the recovery of coliphages using doble layer or single layer methods, but such methods should be evaluated by means of controls, before applying them directly in the analysis of water samples.

Este artículo valora las diferencias en la medición entre laboratorios de siete biomarcadores de un estudio poblacional de adultos mayores en Costa Rica, denominado CRELES. Usa los datos de ensayos replicados en varios laboratorios para la misma muestra biológica. Encuentra una alta estabilidad (medida por el coeficiente de correlación) para casi todas las pruebas, sin embargo se observan diferencias significativas en los promedios entre laboratorios, es decir existen sesgos en la medición de los biomarcadores. Los sesgos son especialmente grandes para creatinina y hemoglobina glicosilada. Las diferencias observadas entre los distintos laboratorios refuerzan el concepto de la variabilidad dependiente tanto del analito como del paciente y del método de análisis. Dadas las diferencias encontradas se definieron ecuaciones para ajustar los datos emitidos por dos laboratorios con el fin de que sean comparables con un tercer laboratorio usado como referencia. Las dicotomías en riesgo o no derivadas de los resultados de laboratorio luego del ajuste, no presentan diferencias entre laboratorios.