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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.

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Resumen Objetivo: Analizar el estado y la influencia de la variabilidad climática en la calidad del agua de un sistema de abastecimiento para consumo humano en San José, Costa Rica, captado en cuatro microcuencas del cantón Vásquez de Coronado, en el periodo 2017-2018, para brindar recomendaciones al operador y promover la protección de la población abastecida Métodos: Estudio descriptivo de los parámetros de calidad básicos de agua para consumo humano (Escherichia coli, coliformes fecales, turbidez, conductividad, pH, color, temperatura y cloro residual). Los análisis se realizaron siguiendo el Standard Methods for the Examination of Water and Wastewater. Se delimitaron las zonas de captación utilizando sistemas de información geográfica. Se utilizó estadística descriptiva e inferencial para describir la calidad del agua y su variabilidad espacial y estacional. Resultados: La concentración de coliformes fecales en el agua superficial fue > 103 NMP/100 mL y la turbidez >5 UTN en el 76% de las muestras. En la red de distribución se encontraron concentraciones de cloro residual y turbidez (>5 UTN) por encima de los límites recomendados para agua de consumo humano. Se encontró diferencias significativas en todos los parámetros básicos de calidad de agua entre las distintas zonas de captación (p<0,05 en todos los casos), así como entre épocas climáticas por zona de captación. Conclusión: Estos resultados indican que se está utilizando agua de calidad inadecuada para uso y consumo humano, por lo cual se recomienda un monitoreo intensivo y sectorizado de las fuentes para localizar puntos de contaminación.

Abstract Objective: Analyze the water quality and the influence of the climatic variability in supply system for the human consume in San José, Costa Rica, in four catchment areas in Vasquez of Coronado in the period 2017-2018 to give recommendations to the operator and promote the protection of the supplied population. Methods: Descriptive study of the basic parameters of the drinking water (Escherichia coli, fecal coliforms, turbidity, conductivity, pH, color, temperature, and residual chlorine). The analyses were done following Standard Methods for the examination of Water and Wastewater. The catchment areas were delimited using geographic information systems (GIS). Descriptive and inferential statistic was used to describe the water quality, spatial and seasonal variability. Results: The concentration of fecal coliforms in the catchment areas was >10 3 NMP/100 mL and the turbidity >5 UTN in 76% of the samples. In the distribution system were found concentrations of the residual chlorine and turbidity (>5 UTN) above according to recommended limits for drinking water in the country. The results showed significant differences in all of the basic parameters of the water quality between catchment areas (p<0, 05 in all cases) and between climatic periods for catchment areas. Conclusion: These results indicate that water of inadequate quality is being used for human use and consumption, for which reason intensive and sectorized source monitoring is recommended to locate contamination points.

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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.

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Abstract In Latin America, the disease burden of shigellosis is found to coexist with the rapid and rampant spread of resistance to commonly used antibiotics. The molecular basis of antibiotic resistance lies within genetic elements such as plasmids, transposons, integrons, genomic islands, etc., which are found in the bacterial genome. Integrons are known to acquire, exchange, and express genes within gene cassettes and it is hypothesized that they play a significant role in the transmission of multidrug resistance genes in several Gram-negative bacteria including Shigella. A few studies have described antibiotic resistance genes and integrons among multidrug resistant Shigella isolates found in Latin America. For example, in Brazil, Bolivia, Chile, Costa Rica and Peru, class 1 and class 2 integrons have been detected among multidrug resistant strains of Shigella; this phenomenon is more frequently observed in S. flexneri isolates that are resistant to trimethoprim, sulfamethoxazole, streptomycin, ampicillin, chloramphenicol, and tetracycline. The gene cassette sul2, which is frequently detected in Shigella strains resistant to the sulfonamides, suggests that the sulfonamide-resistant phenotype can be explained by the presence of the sul2 genes independent of the integron class detected. It is to be noted that sul3 was negative in all isolates analyzed in these studies.The high frequency of sulfonamide (as encoded by sul2) and trimethoprim resistance is likely to be a result of the recurrent use of trimethoprim sulfamethoxazole as a popular regimen for the treatment of shigellosis. The observed resistance profiles of Shigella strains confirm that ampicillin and trimethoprim-sulfamethoxazole are ineffective as therapeutic options. In-depth information regarding antibiotic resistance mechanism in this pathogen is needed in order to develop suitable intervention strategies. There is a pressing need for regional and local antimicrobial resistance profiling of Shigella to be included as a part of the public health strategy.

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In Latin America, the disease burden of shigellosis is found to coexist with the rapid and rampant spread of resistance to commonly used antibiotics. The molecular basis of antibiotic resistance lies within genetic elements such as plasmids, transposons, integrons, genomic islands, etc., which are found in the bacterial genome. Integrons are known to acquire, exchange, and express genes within gene cassettes and it is hypothesized that they play a significant role in the transmission of multidrug resistance genes in several Gram-negative bacteria including Shigella. A few studies have described antibiotic resistance genes and integrons among multidrug resistant Shigella isolates found in Latin America. For example, in Brazil, Bolivia, Chile, Costa Rica and Peru, class 1 and class 2 integrons have been detected among multidrug resistant strains of Shigella; this phenomenon is more frequently observed in S. flexneri isolates that are resistant to trimethoprim, sulfamethoxazole, streptomycin, ampicillin, chloramphenicol, and tetracycline. The gene cassette sul2, which is frequently detected in Shigella strains resistant to the sulfonamides, suggests that the sulfonamide-resistant phenotype can be explained by the presence of the sul2 genes independent of the integron class detected. It is to be noted that sul3 was negative in all isolates analyzed in these studies.The high frequency of sulfonamide (as encoded by sul2) and trimethoprim resistance is likely to be a result of the recurrent use of trimethoprim sulfamethoxazole as a popular regimen for the treatment of shigellosis. The observed resistance profiles of Shigella strains confirm that ampicillin and trimethoprim-sulfamethoxazole are ineffective as therapeutic options. In-depth information regarding antibiotic resistance mechanism in this pathogen is needed in order to develop suitable intervention strategies. There is a pressing need for regional and local antimicrobial resistance profiling of Shigella to be included as a part of the public health strategy.(AU)

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In Latin America, the disease burden of shigellosis is found to coexist with the rapid and rampant spread of resistance to commonly used antibiotics. The molecular basis of antibiotic resistance lies within genetic elements such as plasmids, transposons, integrons, genomic islands, etc., which are found in the bacterial genome. Integrons are known to acquire, exchange, and express genes within gene cassettes and it is hypothesized that they play a significant role in the transmission of multidrug resistance genes in several Gram-negative bacteria including Shigella. A few studies have described antibiotic resistance genes and integrons among multidrug resistant Shigella isolates found in Latin America. For example, in Brazil, Bolivia, Chile, Costa Rica and Peru, class 1 and class 2 integrons have been detected among multidrug resistant strains of Shigella; this phenomenon is more frequently observed in S. flexneri isolates that are resistant to trimethoprim, sulfamethoxazole, streptomycin, ampicillin, chloramphenicol, and tetracycline. The gene cassette sul2, which is frequently detected in Shigella strains resistant to the sulfonamides, suggests that the sulfonamide-resistant phenotype can be explained by the presence of the sul2 genes independent of the integron class detected. It is to be noted that sul3 was negative in all isolates analyzed in these studies. The high frequency of sulfonamide (as encoded by sul2) and trimethoprim resistance is likely to be a result of the recurrent use of trimethoprim sulfamethoxazole as a popular regimen for the treatment of shigellosis. The observed resistance profiles of Shigella strains confirm that ampicillin and trimethoprim-sulfamethoxazole are ineffective as therapeutic options. In-depth information regarding antibiotic resistance mechanism in this pathogen is needed in order to develop suitable intervention strategies. There is a pressing need for regional and local antimicrobial resistance profiling of Shigella to be included as a part of the public health strategy.

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La contaminación fecal de las aguas superficiales es un problema importante para la salud pública, dada la transmisión de microorganismos patógenos. Se estima que las poblaciones ubicadas cerca de costas, ríos o lagos con elevada contaminación fecal, tienen mayor riesgo de desarrollar enfermedades infecciosas gastrointestinales. En esta investigación se analizó durante un año, la contaminación fecal en las aguas superficiales de la microcuenca del río Purires, ubicada en una zona de alta densidad poblacional en Costa Rica. En el 100% de las muestras se detectó contaminación fecal, siendo el punto de muestreo 3 el que mostró los niveles más altos, en promedio 2,2 x 10(4) Número Más Probable(NMP)/100()mL. Aunque los puntos 1 y 2 presentaron menor contaminación fecal, en promedio 6,4 x 10² NMP/100 mL y 6,3 x 10³ NMP/100 mL respectivamente, estos valores indican también mala calidad de las aguas. Con estos resultados se pretende llamar la atención sobre la problemática ambiental de alta contaminación fecal en las aguas superficiales de esta microcuenca. Esta información es un insumo para desarrollar acciones de control sobre las fuentes de contaminación que afectan la calidad de las aguas y por ende, la salud de las poblaciones ubicadas en su cercanía.

Fecal contamination of the superficial waters is an important public health problem due to the presence of pathogenic microorganisms- It has been estimated that populations located close to coasts, rivers of lakes with an elevated fecal contamination have a higher risk of developing communicable gastrointestinal diseases. This investigation analyzed the fecal contamination of the superficial waters of the microbasin of the Rio Purires, located in a highly populated area of Costa Rica. Fecal contamination was detected in 100% of the samples, and sample point 3 was the one that showed the highest levels, with an average of 2.2 x 10(4) Most Probable Number (MPN)/100mL. Even thought points 1 and 2 presented a lower fecal contamination, an average of 6.4 x 10² MPN/100mL and 6.3 x 10³ MPN/100mL, respectively, these values also indicate low quality water. With these results we intend to alert regarding the problem of a high fecal contamination of superficial waters of this microbasin. This information is an input for developing control actions over the contamination sources which compromise the quality of water and, therefore, the health of the populations located in its vicinity.

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Se analizó la calidad microbiológica en 37 muestras de lechuga variedad criolla (Lactuca sativa var. Capitata L.) de distintos intermediarios en las provincias de San José y Cartago, en Costa Rica. Las muestras se recolectaron mediante muestreo no probabilístico por selección intencional. Se cuantificó Escherichia coli (NMP/g) como indicador de contaminación fecal y se determinó la presencia de patógenos específicos (Shigella y Salmonella), por cultivo y por PCR-Múltiple. En el 65% de las muestras analizadas se detectó E. coli, aunque no se encontró Shigella ni Salmonella por PCR-Múltiple o cultivo. Una posible explicación es que los niveles de contaminación de Shigella y Salmonella están por debajo de los límites de detección de ambos métodos (menos de 10(4) UFC/g para Shigella y menos de 10² UFC/g para Salmonella). Estos resultados establecen una base importante para continuar con este tema de investigación y analizar otras fuentes de transmisión de Shigella y Salmonella, dado que ambos patógenos son frecuentes en la región.

The microbiological quality of 37 lettuce samples of the creole variety (Lactuca sativa var. Capitata L.) obtained from different intermediaries at the provinces of San José and Cartago, in Costa Rica was analyzed. The samples were collected through a non-probabilistic sampling with intentional selection. Escherichia coli (NMP/g) was quantified as indicator of fecal contamination and the presence of specific pathogens (Shigella and Salmonella) was determined by culture and Multiplex-PCR. In 65% of the samples analyzed we detected E. coli, even though we did not find Shigella or Salmonella by Multiplex-PCR or culture. A possible explanation is that the Shigella or Salmonella contamination levels may have been under the detection limits for both methods (less than 10(4) CFU/g for Shigella, and less than 10² CFU/g for Salmonella). These results establish an important basis for continuing with this research subject and analyzing other sources of transmission of Shigella and Salmonella contamination, since both pathogens are frequent in the region.

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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 10(7) CFU/ml). DNA was extracted directly from lettuce after inoculation (direct-PCR) and after an enrichment step (enrichment PCR). Multiplex PCR detection limit was 10(4) CFU/ml, diagnostic sensitivity and specificity were 100 percent 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 10(6) CFU/ml, diagnostic sensitivity was 53 percent and diagnostic specificity was 100 percent. 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.

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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 10(7) CFU/ml). DNA was extracted directly from lettuce after inoculation (direct-PCR) and after an enrichment step (enrichment PCR). Multiplex PCR detection limit was 10(4)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 10(6) 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.