Novel members of the order Picornavirales identified in freshwater from Guarapiranga reservoir in São Paulo.

The global challenge of water resource availability is exacerbated by anthropogenic influences that promote the emergence of pollutants. Among these pollutants are microbiological agents, including viruses, which are ubiquitous in the biosphere and play a pivotal role in both ecological balance and the occurrence of diseases in animals and plants. Consequently, monitoring viruses in water sources becomes indispensable for the establishment of effective prevention, promotion, and control strategies. Within this context, the study focuses on the identification of novel viruses belonging to the Picornavirales order in freshwater from the Guarapiranga Reservoir in the state of São Paulo, Brazil. The samples were subjected to viral metagenomics. Our analysis led to the characterization of four distinct sequences (GinkV-05, AquaV_10, MarV_14, and MarV_64), which exhibited significant divergence compared to other members of the Picornavirales order. This remarkable diversity prompted the identification of a potential new genus within the Marnaviridae family, tentatively named Ginkgonavirus. Additionally, we characterized four sequences in a very distinct clade and propose the recognition of a novel family (named Aquaviridae) within the Picornavirales order. Our findings contribute valuable insights into the previously uncharted diversity of Picornavirales present in water sources, shedding light on an important facet of viral ecology and evolution in aquatic environments.

Spread and persistence of antimicrobial resistance genes in wastewater from human and animal sources in São Paulo, Brazil.

The spread of antimicrobial resistance (AMR) through multiple reservoirs is a global concern. Wastewater is a critical AMR dissemination source, so this study aimed to assess the persistence of resistance genetic markers in wastewater using a culture-independent approach. Raw and treated wastewater samples (n = 121) from a wastewater treatment plant (WWTP), a human hospital, a veterinary hospital, and a pig farm were monthly collected and concentrated by filtration. DNA was extracted directly from filter membranes, and PCR was used in the qualitative search of 32 antimicrobial resistance genes (ARGs). Selected genes (blaCTX-M, blaKPC, qnrB, and mcr-1) were enumerated by quantitative real-time PCR (qPCR). Twenty-six ARGs were detected in the qualitative ARGs search, while quantitative data showed a low variation of the ARG's relative abundance (RA) throughout the months, especially at the human hospital and the WWTP. At the WWTP, despite significantly reducing the absolute number of gene copies/L after each treatment stage (p < 0.05), slight increases (p > 0.05) in the RAs of genes blaCTX-M, qnrB, and mcr-1 were observed in reused water (tertiary treatment) when compared with secondary effluent. Although the increase is not statistically significant, it is worth noting that there was some level of ARGs concentration after the disinfection process. No significant absolute or relative after-treatment quantification reductions were observed for any ARGs at the veterinary hospital or the pig farm. The spread of ARGs through sewage needs to be continuously addressed, because their release into natural environments may pose potential risks of exposure to resistant bacteria and impact local ecosystems.

Cross-continental comparative experiences of wastewater surveillance and a vision for the 21st century.

The COVID-19 pandemic has brought the epidemiological value of monitoring wastewater into sharp focus. The challenges of implementing and optimising wastewater monitoring vary significantly from one region to another, often due to the array of different wastewater systems around the globe, as well as the availability of resources to undertake the required analyses (e.g. laboratory infrastructure and expertise). Here we reflect on the local and shared challenges of implementing a SARS-CoV-2 monitoring programme in two geographically and socio-economically distinct regions, São Paulo state (Brazil) and Wales (UK), focusing on design, laboratory methods and data analysis, and identifying potential guiding principles for wastewater surveillance fit for the 21st century. Our results highlight the historical nature of region-specific challenges to the implementation of wastewater surveillance, including previous experience of using wastewater surveillance, stakeholders involved, and nature of wastewater infrastructure. Building on those challenges, we then highlight what an ideal programme would look like if restrictions such as resource were not a constraint. Finally, we demonstrate the value of bringing multidisciplinary skills and international networks together for effective wastewater surveillance.

Characterization of Escherichia coli Isolated from Sows Presenting Purulent Vulvar Discharge.

Purulent vulvar discharge is a clinical sign of genitourinary tract infections, which are a significant concern in swine facilities, leading to sow culling and mortality. Escherichia coli is one of the main agents involved in these diseases. This study aimed to characterize the virulence and antimicrobial resistance profiles as well as the phylotype of Escherichia coli strains isolated from sows with purulent vulvar discharge. The results showed that at least 2 of the 29 tested virulence genes related to extraintestinal pathogenic E. coli were present in all strains tested. The most frequent gene was iutA, present in all strains, followed by the genes iucD, csgA, iss2, and irp2. Associations between iron uptake genes, genes related to adhesion, attachment, and serum resistance, as well as genes related to toxin release and bacteriocin, were frequent. The most prevalent phylotype was B1 (40.0%), followed by A (18.5%), D (11.9%), C (9.6%), B2 (7.4%), E (4.4%), F (1.5%), and Clade I (0.7%), with B2 being related to highly virulent traits. The strains presented elevated resistance to antimicrobials such as ciprofloxacin, streptomycin, cephalexin, florfenicol, and ampicillin. More than 90% of the strains were identified as multidrug-resistant, indicating the selection that is induced by the high use of antimicrobials in swine farming.

Wastewater-based epidemiology for preventing outbreaks and epidemics in Latin America - Lessons from the past and a look to the future.

Wastewater-based epidemiology (WBE) is an approach with the potential to complement clinical surveillance systems. Using WBE, it is possible to carry out an early warning of a possible outbreak, monitor spatial and temporal trends of infectious diseases, produce real-time results and generate representative epidemiological information in a territory, especially in areas of social vulnerability. Despite the historical uses of this approach, particularly in the Global Polio Eradication Initiative, and for other pathogens, it was during the COVID-19 pandemic that occurred an exponential increase in environmental surveillance programs for SARS-CoV-2 in wastewater, with many experiences and developments in the field of public health using data for decision making and prioritizing actions to control the pandemic. In Latin America, WBE was applied in heterogeneous contexts and with emphasis on populations that present many socio-environmental inequalities, a condition shared by all Latin American countries. This manuscript addresses the concepts and applications of WBE in public health actions, as well as different experiences in Latin American countries, and discusses a model to implement this surveillance system at the local or national level. We emphasize the need to implement this sentinel surveillance system in countries that want to detect the early entry and spread of new pathogens and monitor outbreaks or epidemics of infectious agents in their territories as a complement of public health surveillance systems.

Serotyping and Antimicrobial Susceptibility Profiling of Glaesserella parasuis Isolated from Diseased Swine in Brazil.

Glaesserella parasuis is one of the major pathogens in swine intensive production systems. To date, 15 serovars have been described, and the prevalence of these serotypes in different geographical regions has been identified by several methods. G. parasuis outbreaks could be controlled with vaccination if it were not for serovar diversity and limited cross-serovar protection; consequently, antibiotic therapy continues to be necessary for infection control. Here, we present the isolation, identification, serotyping, and antibiotic susceptibility profiling of G. parasuis from diseased swine in Brazil. A total of 105 G. parasuis strains, originating from nine different Brazilian states, were evaluated, and serotypes 4 and 5 were found to be the most prevalent (27.6% and 24.8% respectively). Aminoglycosides, florfenicol, tiamulin, and ß-lactams were tested, and they presented lower resistant rates against G. parasuis strains. The highest resistance rates were observed against tylosin (97.1%), sulfadimethoxine (89.5%), danofloxacin (80%), trimethoprim/sulfamethoxazole (62.5%), enrofloxacin (54.3%), and clindamycin (50.5%). Multidrug resistance was detected in 89.5% of tested strains, and a total of sixty resistance profiles were identified. The cluster analysis of resistance patterns showed no correlation with the isolation year or G. parasuis serotype.

One-year surveillance of SARS-CoV-2 in wastewater from vulnerable urban communities in metropolitan São Paulo, Brazil.

The current COVID-19 pandemic has emphasized the vulnerability of communities living in the urban outskirts and informal settlements. The lack of reliable COVID-19 case data highlights the importance and application of wastewater-based epidemiology. This study aimed to monitor the COVID-19 trends in four vulnerable urban communities (slums and low-income neighborhoods) in metropolitan São Paulo by assessing the SARS-CoV-2 RNA viral load in wastewater. We analyzed 160 samples from May 2020 to June 2021 with weekly or fortnightly samplings. The samples were ultracentrifuged with glycine elution and quantified by N1/N2 SARS-CoV-2 RT-qPCR. The results of positivity were 100% (Paraisópolis, Heliópolis and Cidade Tiradentes) and 76.9% (Vila Brasilândia). The new case numbers of COVID-19, counted from the onset of symptoms, positively correlated with SARS-CoV-2 N1 viral loads from the two largest communities (p<0.001). SARS-CoV-2 infectivity was tested in Vero E6 cells after concentration with the two techniques, ultrafiltration (Centricon® Plus-70 10 kDa) and sucrose cushion ultracentrifugation, but none of the evaluated samples presented positive results. Next-generation sequencing (NGS) analysis from samples collected in March and August 2021 revealed the presence of the clade 20 J (lineage P.1) belonging to the most prevalent circulating variant in the country. Our results showed that wastewater surveillance data can be used as complementary indicators to monitor the dynamics and temporal trends of COVID-19. The infectivity test results strengthened the evidence of low risk of infection associated with SARS-CoV-2 in wastewater.

Causes of Sow Mortality and Risks to Post-Mortem Findings in a Brazilian Intensive Swine Production System.

The present study was conducted to investigate the risk factors for post-mortem findings and causes of sow mortality. A post-mortem examination and microbiological investigation were conducted on 123 sows from a breeding herd with 15,000 dams. The mortality of spontaneous death in sows occurred mostly in the peripartum period (53%; p < 0.05). The spontaneous deaths were associated with heart failures, hemorrhagic and perforating gastric ulcers, and liver torsion, while in the euthanized sows, the post-mortem findings were associated with locomotor disorders. A higher body condition score (BCS ≥ 3.5) increased (p < 0.05) heart failure on the post-mortem examination. The excessive use of manual obstetric interventions increased sow deaths resulting from cervix/uterus ruptures and increased the odds of death (p < 0.05) due to metritis. Sow mortality had a multifactorial etiology. Infections were polymicrobial. The main microbial agents identified from a septic lesion in locomotor, genitourinary, and respiratory systems were Trueperella pyogenes, Escherichia coli, and Actinobacillus pleuropneumoniae, respectively. In conclusion, sow mortality involved multiple risk factors and several bacterial agents. These results indicate that better management practices can reduce sow mortality in swine production and increase sow welfare.

Pandemic Clones of CTX-M-15 Producing Klebsiella pneumoniae ST15, ST147, and ST307 in Companion Parrots.

Psittacine birds are commonly kept as companion birds and the maintenance of these birds in captivity may represent a zoonotic risk and contribute to the propagation of multidrug-resistant and ß-lactamase extended-spectrum (ESBLs)-producing pathogens. This study aimed to identify and characterize strains of the Klebsiella pneumoniae complex isolated from diseased psittacine birds, determining virulence and resistance profiles. K. pneumoniae strains were isolated from 16 birds (16/46). All strains carried more than three virulence genes, with a high frequency of fimH and kpn (93.75%), uge (87.52%), and irp-2 (81.25%) genes. The antimicrobial susceptibility revealed that 3/16 strains were ESBL producers. Genomic analysis revealed that CTX-M-15-positive strains belonged to sequence types (STs) ST15, ST147, and ST307, characterized as international clones associated with outbreaks of healthcare-associated infections (HAIs) worldwide.

Vaginal microbiota signatures in healthy and purulent vulvar discharge sows.

Purulent vulvar discharges, primarily caused by genito-urinary tract infections, are an important source of economic loss for swine producers due to sow culling and mortality. However, the agents that compose the vaginal microbiota of sows and their changes during infections are not well understood. The first goal of this study was to characterize and compare the vaginal bacterial content of healthy (HE, n = 40) and purulent vulvar discharge sows (VD, n = 270) by a culture-dependent method and MALDI-TOF MS identification. Secondly, we performed 16S rRNA targeted metagenomic approach (n = 72) to compare the vaginal microbiota between these groups. We found a wide variety of bacteria, with Proteobacteria, Firmicutes, and Bacteroidota being the most abundant phyla in both groups, as well as Escherichia-Shigella, Streptococcus, and Bacteroides at the genus level. Most agents identified in the sequencing method also grew in the culture-dependent method, showing the viability of these bacteria. Alpha diversity did not differ between HE and VD sows, regarding sample richness and diversity, but a beta-diversity index showed a different microbiota composition between these groups in two tested herds. ANCOM analysis revealed that Bacteroides pyogenes were more abundant in VD females and can be a marker for this group. Other agents also require attention, such as the Streptococcus dysgalactiae and Staphylococcus hyicus found in remarkably greater relative abundance in VD sows. Network analysis revealed important positive correlations between some potentially pathogenic genera, such as between Escherichia-Shigella, Trueperella, Streptococcus, Corynebacterium, and Prevotella, which did not occur in healthy sows. We conclude that the alteration of the vaginal microbiota between healthy and purulent vulvar discharge sows, although not extreme, could be due to the increase in the relative abundance of specific agents and to associations between potentially pathogenic bacteria.

Hepatitis A Outbreaks and Environmental Circulation of Genotype IA Strains in the São Paulo City, 2017-2018.

Hepatitis A virus (HAV) is the major cause of enterically transmitted infectious hepatitis. Between 2016 and 2017, the number of confirmed cases of hepatitis A virus (HAV) increased from 64 to 786 in São Paulo affecting mainly adults aged between 18 and 39 years (80%) and males (88%). To support epidemiological surveillance, the present study monitored the presence of HAV in urban sewage samples collected bimonthly for 1 year (November 2017-November 2018) in the central region of the city, where most of cases were detected. Sewage samples were concentrated by polyethylene glycol precipitation and HAV RNA was quantified by RT-qPCR. Nucleotide sequencing targeting the VP1/2A junction region was carried out to genotype the HAV strains. HAV was detected in 76.9% (40/52) of the samples, with a geometric mean viral load of 5.09 × 104 (± SD 4.51 × 105) genome copies (GC/L) (Mauá Street) and 5.27 × 104 (± SD 1.26 × 106) GC/L (Prestes Maia Avenue). Of the 40 positive samples, 8 were typed as HAV subgenotype IA [100% nucleotide (nt) identity with HAV strain VRD_521_2016]. Highest homology was obtained with sequences from European countries (Italy, Spain) and Israel, all of which had reported recent HAV outbreaks associated with men who have sex with men. Our results highlight that wastewater surveillance is a useful tool to support investigating HAV outbreaks in the community, including circulating genotypes.

Main bacterial species causing clinical disease in ornamental freshwater fish in Brazil.

Bacterial diseases are common in ornamental fish, more frequently associated with ubiquitous bacteria from the aquarium environment. The disease can lead to fish mortality and cause high economic losses if not rapidly controlled. The aim of this study was to identify the main causative bacterial agents of infection in ornamental fish with different clinical signs. A total of 126 freshwater fish, from 12 families and 38 species, with clinical signs were collected in a wholesaler in São Paulo, SP, Brazil. Samples were taken from the eye, skin ulcers, kidneys, and gills, plated on MacConkey, CHROMagar Orientation, and blood agar and incubated under aerobic and anaerobic conditions. Bacterial identification was performed by MALDI-TOF mass spectrometry. From the 126 studied animals, 112 were positive for bacterial isolation. Among the positive animals, 32.1% presented infection caused by a single bacterial species, while in the remaining 67.9%, two to six different bacterial species were identified. A total of 259 bacterial strains were obtained and classified among 46 bacterial species. The species of higher frequency were Aeromonas veronii (26.3%), Aeromonas hydrophilla (16.2%), Shewanella putrefaciens (7.3%), Citrobacter freundii (8.1%), Vibrio albensis (5.8%), and Klebsiella pneumoniae (4.2%). MALDI-TOF MS showed to be a rapid method for diagnosis of bacterial disease outbreaks in ornamental fish establishments.

Vagococcus sp. a porcine pathogen: molecular and phenotypic characterization of strains isolated from diseased pigs in Brazil.

INTRODUCTION: Vagococcus spp. is known for its importance as a systemic and zoonotic bacterial pathogen even though it is not often reported in pigs. This is related to the pathogen misidentification due to the lack of usage of more discriminatory diagnostic techniques. Here we present the first report of Vagococcus lutrae in swine and the characterization of Vagococcus fluvialis and Vagococcus lutrae isolated from diseased animals. METHODOLOGY: Between 2012 and 2017, 11 strains with morphological characteristics similar to Streptococcus spp. were isolated from pigs presenting different clinical signs. Bacterial identification was performed by matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry and confirmed by 16S rRNA sequencing and biochemical profile. Strains were further genotyped by single-enzyme amplified fragment length polymorphism (SE-AFLP). Broth microdilution was used to determine the minimal inhibitory concentration of the antimicrobials of veterinary interest. RESULTS: Ten strains were identified as V. fluvialis and one was identified as V. lutrae. The SE-AFLP analysis enabled the species differentiation with specific clustering of all V. fluvialis separately from the V. lutrae strain. Most strains presented growth in the maximum antibiotic concentration values tested for eight of the 10 analyzed antimicrobial classes. CONCLUSIONS: The observed resistance pattern can represent a problem for veterinary and producers in the treatment of diseases associated Vagococcus spp. in swine production. Vagococcus species may also be a risk for pig industry workers. The data described here will be of great value in further understanding the behavior of this pathogen in animal production.

Survival of thermotolerant coliforms in municipal biosolids after application in tropical soil cultivated with Eucalyptus.

The increasing production of biosolids in urban areas has been led to a search for alternative disposal avenues of this residue, which is rich in organic matter and nutrients. Agricultural land application of biosolids, motivated by its fertilizing power, is a widespread practice in many countries, but there are safety and regulatory concerns regarding the presence of pathogens in Class B biosolids. In addition, the scarcity of studies under tropical climate conditions raises questions that impede the agricultural use of this residue in some regions. The objective of this study was to evaluate the survival of thermotolerant coliforms over 12 months (52 weeks) after biosolids application on the surface of a Quartzipsamment neosol cultivated with Eucalyptus. Two different biosolids were studied: those generated by a biological treatment system with complete mixing aeration ponds followed by decantation ponds (Treatment A) and biosolids from a biological treatment system with conventional activated biosolids reactors (Treatment B), both delineated in randomized blocks with four replicates. After application on the forest soil, we estimated an average survival time of 54 weeks for thermotolerant coliforms present in Treatment A biosolids and 93 weeks in Treatment B biosolids. Thermotolerant coliforms persist much longer under tropical climate conditions in Brazil than in comparable studies under temperate climate conditions. This reaffirms the need to carry out studies covering the full range of moisture and temperature regimes in which biosolids are applied as fertilizer.

Assessment of health risks from recreational exposure to Giardia and Cryptosporidium in coastal bathing waters.

Some Brazilian beaches are impacted by raw or poorly treated sewage. Thus, users (beachgoers, sports people, and children) are exposed to pathogens, which pose health concerns. This study aimed to estimate the probability of infection and disease by Giardia and Cryptosporidium, using the quantitative microbial risk assessment (QMRA), for three groups of bathers: children, adults, and open water swimmers. The concentrations of (oo)cysts were taken from a study run by CETESB (Environmental Company of Sao Paulo State) throughout 2011 and 2012, in which 203 samples were collected monthly and analyzed for (oo)cysts of Giardia and Cryptosporidium. Giardia was present in 43% of the samples, while Cryptosporidium in 13%. Infection probability was higher in beaches with more positive samples for Giardia cysts for the group of open water swimmers. In some cases, the highest annual risk obtained for giardiasis and cryptosporidiosis was 2.4 × 10-1 and 8.9 × 10-3 for open water swimmers, respectively, exceeding the incidence results found in the epidemiological study run in summer of 1999 in São Paulo state coast. The results bring insights to improve environmental quality in order to protect tourists' and residents' wellbeing.

Companion psittacine birds as reservoir of gentamicin and vancomycin-resistant Enterococcus spp / Psitacídeos de companhia como reservatórios de Enterococcus spp. gentamicina e vancomicina-resistentes

Enterococcus are recognized worldwide as significant nosocomial agents that have been continuously envolving to adapt to different niches and acquire resistance to several antibiotic classes. Vancomycin and gentamicin-resistant strains of E. faecalis and E. faecium have been associated with nosocomial human infections. Some epidemiological studies suggest the participation of pets as reservoirs of vancomycin and gentamicin-resistant Enterococcus strains. However, the role of companion birds as reservoirs of these strains has been poorly studied. In this study, 126 psittacine birds were evaluated and 26.9% carried Enterococcus spp., including the species E. faecalis, E. faecium, E. hirae, E. phoeniculicola, E. gallinarum and E. casseliflavus. The antibiotic resistance profile showed four high-level gentamicin-resistance (HLGR) strains. In addition, two strains presented intermediate levels of vancomycin resistance. Resistant strains were isolated from fecal and oropharynx samples of sick and clinically healthy birds, suggesting that psittacine birds may act as reservoirs of HLGR Enterococcus spp. However, sick birds appear to be more implicated in the enterococci transmission than healthy birds.(AU)

Enterococcus são reconhecidos mundialmente como significantes agentes nosocomiais, que têm continuamente se adaptado a diferentes nichos e adquirido resistência a várias classes de antibióticos. Cepas de E. faecalis e E. faecium vancomicina e gantamicina-resistentes têm sido associadas a infecções nosocomiais em humanos. Alguns estudos epidemiológicos sugerem a participação de aves como reservatórios de cepas de Enterococcus vancomicina e gentamicina-resistentes. Entretanto, a relação das aves de companhia como reservatórios destas cepas tem sido pouco estudada. Neste estudo, 126 psitacídeos foram avaliados, e 26,9% destes eram portadores de Enterococcus spp., incluindo as espécies E. faecalis, E. faecium, E. hirae, E. phoeniculicola, E. gallinarum e E. casseliflavus. O perfil de resistência antibiótica mostrou quatro cepas com alto nível de resistência a gentamicina (ANRG). Além de duas cepas com nível intermediário de resistência a vancomicina. As cepas resistentes foram isoladas de amostras fecais e de orofaringe de aves doentes e clinicamente saudáveis, sugerindo que psitacídeos podem estar atuando como reservatórios para Enterococcus spp. com ANRG. Contudo, Aves doentes parecem estar mais relacionadas à transmissão de enterococcus, do que aves saudáveis.(AU)

A critical analysis of recreational water guidelines developed from temperate climate data and applied to the tropics.

Recreational water epidemiology studies are rare in settings with minimal wastewater treatment where risk may be highest, and in tropical settings where warmer temperature influences the ecology of fecal indicator bacteria commonly used to monitor recreational waters. One exception is a 1999 study conducted in São Paulo Brazil. We compared the risk and exposure characteristics of these data with those conducted in the United Kingdom (UK) in the early 1990s that are the basis of the World Health Organization's (WHO) guidelines on recreational water risks. We then developed adjusted risk difference models (excess gastrointestinal illness per swimming event) for children (<10 years of age) and non-children (≥10 years of age) across five Brazil beaches. We used these models along with beach water quality data from 2004 to 2015 to assess spatial and temporal trends in water quality and human risk. Risk models indicate that children in Brazil have as much as two times the risk of gastrointestinal illness than non-children. In Brazil, 11.8% of the weekly water samples from 2004 to 2015 exceeded 158 enterococci CFU/100 ml, the highest level of fecal streptococci concentration measured in the UK study. Risks associated with these elevated levels equated to median NEEAR-Gastrointestinal Illness (NGI) risks of 53 and 96 excess cases per 1000 swimmers in non-children and children, respectively. Two of the five beaches appear to drive the overall elevated NGI risks seen during this study. Distinct enteric pathogen profiles that exist in tropical settings as well as in settings with minimal wastewater treatment highlight the importance of regionally specific guideline development.

Performance of wastewater reclamation systems in enteric virus removal.

Analysis of virus removal by tertiary or advanced sewage treatment processes is an emerging topic due to importance of reusing water on a global level. This study aimed to monitor important human viral pathogens: the human adenovirus (HAdV), JC polyomavirus (JCV) and Species A rotaviruses (RVA) in urban sewage, secondary effluents and reclaimed water from metropolitan São Paulo (MSP), Brazil. Four large wastewater treatment plants (WWTPs) in MSP were sampled monthly during a one-year period (April 2015 to March 2016). The viruses were quantified by quantitative PCR (qPCR), and HAdV viability was tested by the integrated cell culture (ICC)-qPCR assay. WWTPs are composed of activated sludge processes and different tertiary treatments (coagulation/sedimentation, sand-anthracite filters, membrane bioreactors (MBRs)/reverse osmosis (RO) and disinfection by chlorination). Physicochemical parameters were also evaluated to verify association with density of viruses detected in different treatment stages. HAdV, JCV and RVA were consistently detected (100%) in the sewage influent samples (range: 106-108 genome copies GC/L). In the secondary effluent, HAdV was detected in 100% (48/48) of the analysed samples, JCV in 85.4% and RVA in 97.9% (range: 104-107 GC/L for all viruses tested). HAdV was the most frequently detected virus in the tertiary effluent (62.2%) (28/45), exhibiting a viability between 0 and 44% of the tested samples in the wastewater reclamation systems. The MBR/RO systems demonstrated better virus removal efficiencies (range: 2.3-2.9 log10). Temperature, pH, turbidity and total organic carbon presented association with the viral density in the reclaimed water samples. Presence of viruses in treated effluents can indicates health risks depending on uses of recovery water. Further risk assessment studies should be conducted to better assess health risks under different exposure scenarios for water recovery in urban settings.

Persistence of Fecal Contamination Indicators and Pathogens in Class B Biosolids Applied to Sugarcane Fields.

Agricultural recycling of human Class B biosolids in sugarcane ( spp.) crop is a promising alternative to reduce the costs of biosolids disposal. However, the presence of fecal contamination indicators such as thermotolerant coliforms and pathogenic organisms such as enterovirus and spp. in biosolids impose barriers to effective and widespread use of biosolids as fertilizer. In addition, there is a scarcity of studies that investigate the persistence of these organisms in tropical soils. This study aimed to evaluate the persistence of pathogenic and fecal indicators for 258 d in a tropical clayey soil amended with human Class B biosolids and cultivated with sugarcane. Treatments were immediate incorporation of biosolids into soil after application (T1) or superficial application of biosolids followed by incorporation after 35 d (T2), emulating the typical procedure in sugarcane fields. Thermotolerant coliforms were estimated to persist for 437 d in T1 and 398 d in T2. For enterovirus, mean estimated persistence time in soil was 26 d for T1, but the sampling frequency was insufficient in T2 for persistence analysis. After 35 d, no enterovirus was detected in any sample. Mean estimated persistence time for viable spp. eggs in soil was 22 d in T1 and 41 d in T2.