The continuous introduction of cleaning products containing benzalkonium chloride (BAC) from household discharges can mold the microbial communities in wastewater treatment plants (WWTPs) in a way still poorly understood. In this study, we performed an in vitro exposure of activated sludge from a WWTP in Costa Rica to BAC, quantified the changes in intI1, sul2, and qacE/qacEΔ1 gene profiles, and determined alterations in the bacterial community composition. The analysis of the qPCR data revealed elevated charges of antibiotic resistance genes in the microbial community; after BAC's exposure, a significant increase in the qacE/qacEΔ1 gene, which is related to ammonium quaternary resistance, was observed. The 16S rRNA gene sequences’ analysis showed pronounced variations in the structure of the bacterial communities, including reduction of the alpha diversity values and an increase of the relative abundance of Alphaproteobacteria, particularly of Rhodospseudomonas and Rhodobacter. We confirmed that the microbial communities presented high resilience to BAC at the mg/mL concentration, probably due to constant exposure to this pollutant. They also presented antibiotic resistance-related genes with similar mechanisms to tolerate this substance. These mechanisms should be explored more thoroughly, especially in the context of high use of disinfectant.

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.

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 the 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 (WWTP) in Costa Rica were analyzed for somatic coliphage and human enterovirus, hepatitis A virus, norovirus genotype I and II, and rotavirus. All WWTP 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 plaque forming unit per 100 mL, which corresponded to an increased likelihood of encountering enteric viruses above the limit of detection (>1.83×102 virus target/100 mL). Additionally, quantitative microbial risk assessment was executed for famers 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.

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.

Specific serum antibody titres to defined lipopolysaccharides (LPS) of Shigella spp. and Salmonella serogroup B (BO) and Shigella invasion plasmid antigens (Ipa) were determined by enzyme immunoassays in healthy Costa Rican women from low (n = 34), middle (n = 36) and high (n = 19) socioeconomic conditions and from 64 Swedish women. Specific IgG antibody titres were the highest, in particular to the S. flexneri Y LPS, with mean titres of 750 (SD = 360), 690 (380), and 820 (300) for the Low, Middle and High Costa Rican groups respectively. Lower titres were recorded against S. sonnei and S. dysenteriae type 1. Titres against Salmonella LPS were very low (< 250). In Swedish women, significantly lower serum anti-LPS titres were found (p < 0.05). No statistically significant differences regarding socioeconomic conditions or serum anti-LPS titres were seen in the Costa Rican groups (p > 0.05). Mean IgG-titres to the Ipa of 330 (150), 220 (170) and 140 (110) were found for the 3 Costa Rican groups, respectively. The IgG and IgM titres to Ipa in serum from the Low group were significantly higher than those of the High group (p < 0.05). The mean serum IgG titre of 70 (90) for the Swedish women was significantly lower than that of the Low and Middle groups (p < 0.05), but similar to that of the High group (p > 0.05). High IgG anti-Ipa titres were found in 38% from the Low, 19% from the Middle but none in the High group (mean value of the High group +2 SD).(ABSTRACT TRUNCATED AT 250 WORDS)