Microcosm environment models for studying the stability of adenovirus and murine norovirus in water and sediment.

Microcosms are useful tools for understanding the survival and fate of enteric viruses in aquatic environments. This study set out to determine the stability of infectious enteric viruses in an aquatic environment using a laboratory scale microcosm. Sediment and overlaying water were collected from a lagoon and inoculated with known concentrations of recombinant adenovirus (AdV-GFP) and murine norovirus (MNV-1). Infectious particles of these viruses were measured using fluorescence microscopy (AdV-GFP) or the plaque assay method (MNV-1), over 85 days in two different conditions: under natural sunlight and in fully darkened environments. The time required to reach one log reduction in viral titres (T90) of viable viruses in a natural condition microcosm for AdV-GFP and MNV-1 was shorter than in a dark condition microcosm. There was also a negative correlation between the temperature and infectivity of these viruses in both water and sediment samples. Considering that microcosms aim to mimic natural environment conditions and that AdV-GFP and MNV-1 are excellent surrogates for measuring the infectivity of the respective viruses strains, the results presented here have the potential to be applied in future health hazard studies and also would be useful for future climate scenarios.

Enterobacteria associated with houseflies (Musca domestica) as an infection risk indicator in swine production farms.

Houseflies (Musca domestica) spend part of their life development on animal or human manure. Manure is high in pathogenic microbes; thus, houseflies have been known as a mechanical vector for various important zoonotic diseases. Therefore, the present study showcases captured houseflies from intensive swine production regions (which are areas of high manure concentration) in Southern Brazil, and analyses their bodies' to the presence of Escherichia coli and Salmonella sp. and the sensitivity of these bacteria to various antibiotics. Additionally, Quantitative Microbiology Risk Assessment was performed simulating the contamination of lettuce by flies' bacteria and subsequent lettuce consumption by an adult human being. Houseflies were captured in swine buildings and farm houses from five farms. E. coli quantification values ranged from 104 to 106 CFU/20 flies, and all sampling sites had positive results from bacteria presence in the collected houseflies. On the other hand, Salmonella sp. presence was observed in only three farms, where the quantification ranged from 102 to 105 CFU/20 flies. The bacteria showed to be resistant to at least two from the four tested antibiotics (ampicillin, Cefalotin, Ciprofloxacin and Norfloxacin) antibiotics used in human or veterinary medicine. Infection probability analyses showed risk of human infection by E.coli, indicating possible transmission of zoonotic pathogens through flies. In this context, it was possible to conclude that there is a need for flies control, especially in swine farms where zoonotic pathogens can be abundant, to minimize the health impact of the vectorization of enteric bacteria.

Spatial distribution of enteric viruses and somatic coliphages in a Lagoon used as drinking water source and recreation in Southern Brazil.

This study aimed to evaluate the contamination level of the Peri Lagoon, the main freshwater reservoir of Santa Catarina Island, Southern Brazil, for human adenovirus (HAdV), hepatitis A virus (HAV), rotavirus species A (RVA), and somatic coliphages (SOMCPH). Viruses were also investigated in sediments and their sensitivity against natural sunlight was analysed by studying their spatial distribution in different depths of the water column. A total of 84 water samples and 48 sediment samples were examined by qPCR or RT-qPCR. Infectivity of HAdV and SOMCPH was determined and quantified by plaque assay method. A sum of 64% and 48% of water and sediment samples were positive for HAdV, respectively. RVA was present in 33% and 18% of water and sediment samples, and 25% of water samples were positive for HAV. HAdV were infectious in 76% of water and 83% of sediment samples that were positive by qPCR. SOMCPH could be detected in 42% and 18% of water and sediment samples, respectively. The data pointed a variation of viruses' prevalence according to the different water column depths. These results demonstrated that water sources and sediments contaminated by human wastes could play an important role in the recontamination of water columns harvested for further treatment or used for recreational purposes. These data can be of great value for future risk assessment analysis.

Settling and survival profile of enteric pathogens in the swine effluent for water reuse purpose.

The present study evaluated the pathogens persistence and settling profile in swine effluent. We determined the enteric pathogens settling characteristics, their survival and inactivation profile in swine effluent (for water reuse purpose) and in sludge (generated after aerobic treatment - during secondary settling process). The study was performed in laboratorial-scale and in full-scale (manure treatment plant). Enteric viruses and enteric bacteria were used as biomarkers. Results showed that these enteric pathogens were significantly reduced from swine effluent during secondary settling process, and enteric viruses removal was correlated with the suspended solids decantation. The design of secondary settlers can be adapted to improve pathogens removal, by diminishing the solids loading rate per area and time, ending in higher hydraulic retention times.

Waterborne diseases classification and relationship with social-environmental factors in Florianópolis city - Southern Brazil.

This study aimed to investigate and classify the occurrence of waterborne diseases in Florianópolis city, Santa Catarina State, Southern Brazil and to correlate these diseases with the following social-environmental indicators of the local population: type of water supply, adequate collection and sewage treatment, areas of flooding and domestic water tank cleaning. Reports of outpatients were analyzed for surveillance of waterborne diseases during the period of 2002 to 2009. Waterborne diseases were classified into four groups: Group A: diarrheal diseases; Group B: parasitological diseases; Group C: skin diseases and Group D: eye diseases. The diarrheal, parasitological and skin diseases were the most frequently reported. Waterborne diseases belonging to Group A in all sites were correlated with other waterborne diseases groups, which can be an indicator of the circulation of other waterborne diseases. Regarding the social-environmental indicators assessed, the most correlated with waterborne diseases were the origin and quality of the water supply, followed by inadequate collection and treatment of sewage, frequent flooding, and finally the lack of cleanliness of the water reservoir. The results highlight the need for policies aiming for improvement of the sanitation service in the maintenance of human, animal and environmental health.

Enteric viruses in surface water and sediment samples from the catchment area of Peri Lagoon, Santa Catarina State, Brazil.

This paper aims to quantify human adenovirus (HAdV), rotavirus species A (RVA), and hepatitis A virus (HAV) in surface water and sediments and to determine the viability of HAdV in these samples. Water and sediment samples were collected, and HAdV, RVA, and HAV were quantified by real-time polymerase chain reaction (PCR); HAdV was also evaluated for infectivity by a plaque assay (PA). For the water samples, HAdV was detected in 70.8% of the summer collections, with 82.4% containing infectious HAdV; the HAdV incidence in winter was 62.5%. For the sediment samples, the incidence of HAdV was 37.5% in the summer collections, with 66.7% containing infectious HAdV; the HAdV incidence in winter was 37.5%. RVA was detected in 20.8 and 45.8% of surface water samples collected in summer and winter, respectively, and 8.3 and 12.5% of sediment samples collected in summer and winter, respectively. HAV was detected only in surface waters, with 54.8 and 12.5% positivity in summer and winter samples, respectively. This study demonstrated that enteric viruses are present in water and sediments and that the presence of infectious viruses should be investigated whenever possible for quantitative microbial risk assessment studies. Combined analyses of water and sediments are important for reliable public health risk analysis of recreational and lagoon waters.

Utility of specific biomarkers to assess safety of swine manure for biofertilizing purposes.

Swine production is an important economic activity in Brazil, and there is interest in the development of clean production mechanisms to support sustainable agro-industrial activities. The biomass derived from swine manure has good potential to be used as a biofertilizer due to its high nutrient concentration. However, the land application of manure should be based on safety parameters such as the presence of pathogens that can potentially infect animals and people. This study was designed to assess the presence of porcine circovirus-2 (PCV2), porcine adenovirus (PAdV), rotavirus-A (RV-A) and Salmonella spp. in liquid manure, as well the infectivity of two genotypes of circovirus-2 (PCV2a and PCV2b) present in liquid manure. Three swine farms were evaluated: 1) a nursery production farm (manure analyzed before and after anaerobic biodigestion), 2) a grow-finish production farm (analyzed before and after anaerobic biodigestion), and 3) a second grow-finish production farm (raw manure-affluent). PCV2, PAdV and RV-A were present before and after anaerobic biodigestion (either affluent or effluent) at all farms. Salmonella spp. were detected at farm 1 (affluent and effluent) and farm 3 (raw manure-affluent) but not farm 2 (affluent and effluent). When the ability of the anaerobic biodigestion process to reduce viral concentration was evaluated, no significant reduction was observed (P>0.05). Both the PCV2a and PCV2b genotypes were detected, suggesting viral co-infection in swine production. The results revealed infectious PCV2 even after anaerobic biodigestion treatment. The presence of Salmonella spp. and enteric viruses, especially infectious PCV2, in the final effluent from the anaerobic biodigester system suggests that the process is inefficient for pathogen inactivation. Due to the prevalence and infectivity of PCV2 and considering the successful use of molecular methods coupled to cell culture for detecting infectious PCV2, we suggest that this virus can be used as a bioindicator in swine manure treatment systems to check the efficiency of pathogen inactivation and ensure the production of safe biofertilizers from swine manure.

Surveillance of human viral contamination and physicochemical profiles in a surface water lagoon.

The present study evaluated the contamination of a surface water lagoon (Peri Lagoon) in Florianópolis, Santa Catarina, Brazil, by human adenovirus (HAdV), polyomavirus JC (JCPyV), hepatitis A virus (HAV) and rotavirus species A (RVA). Efforts were driven to determine the correlation between viral presence and the physicochemical parameters of the lagoon and measure the distribution of these viruses throughout the year (June 2010 to May 2011). A total of 48 samples were collected, concentrated and analyzed by qPCR (quantitative polymerase chain reaction). Approximately 96% of the samples were positive for HAdV (46/48), 65% were positive for RVA (31/48), 21% were positive for JCPyV (10/48) and 12% were positive for HAV (6/48). The presence of JCPyV was positively correlated with that of NO(2)(-)N, and also there was a positive correlation between the presence of each one of the viruses (HAdV, HAV and RVA) in winter. Samples from water dedicated for human consumption and recreation tested positive for HAdV by qPCR. These samples were also subjected to viral integrity and viability assays: 83% (10/12) contained intact viral particles and 66% (8/12) contained infectious particles. Our results demonstrate the release of human waste into water sources, justifying the urgent need to add viral parameters to water quality surveillance.