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.

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.

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.

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.

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.

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.

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.

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.

Real-time PCR detection of Toxoplasma gondii in surface water samples in São Paulo, Brazil.

Water is considered an important vehicle for the spread of human toxoplasmosis in several countries. Toxoplasma gondii oocysts can persist in the environment for long periods, being highly resistant to the various chemical inactivation processes commonly used by water supply systems, distinctly from simple filtration and flocculation that are efficient in removing oocysts from drinking water. The existing methodologies for identification and quantification of this parasite in water samples are not standardized and have limitations. This study aimed to evaluate the presence of T. gondii oocysts in surface water samples used as a source for the production of drinking water in the State of São Paulo, through the implementation of a specific methodology using real-time PCR technique (qPCR). Volumes of 20 L of the sample were concentrated by filtration in Envirocheck® HV capsules. For DNA extraction, the PowerSoil DNA isolation® kit (currently DNeasy PowerSoil®) was used. The target sequence selected for qPCR was a 62-base-pair fragment of the B1 gene. In the initial recovery evaluation of the method in four replicates of reverse osmosis water, the mean recovery was 48.5% (SD ± 11.5), while the mean recovery for method performance in matrices was 3.2% (SD ± 3.2) (rainy season) and 62.0% (SD ± 6.2) (dry period), suggesting that the characteristics of the samples and the climatic conditions interfere in the recovery efficiency. Of the 39 samples analyzed (May to December 2015), 7.7% (3/39) were positive for T. gondii, and among the ten sources studied; the occurrence of the oocysts was detected in 30% (3/10).

Noroviruses in raw sewage, secondary effluents and reclaimed water produced by sand-anthracite filters and membrane bioreactor/reverse osmosis system.

The importance of noroviruses (NoVs) in the epidemiology of waterborne diseases has increased globally in the last decades. The present study aimed to monitor genogroup I and II noroviruses in different treatment stages of four wastewater treatment plants (WWTPs) in the metropolitan São Paulo. WWTPs consist of secondary (activated sludge) and tertiary treatments (coagulation, sand-anthracite filters, membrane bioreactor (MBR)/reverse osmosis (RO) and chlorination). Raw sewage (500mL) and treated effluents (1L) were concentrated by celite and reclaimed water (40L) by hollow-fiber ultrafiltration system. Quantitative (qPCR) and nested PCR with nucleotide sequencing were used for quantification and molecular characterization. NoVs were widely distributed in raw wastewater samples (83.3%-100% NoV GI and 91.6%-100% NoV GII) and viral loads varied from 3.8 to 6.66log10gcL-1 for NoV GI and 3.8 to 7.3log10gcL-1 for NoV GII. Mean virus removal efficiencies obtained for activated sludge processes ranged from 0.3 to 0.8 log10 for NoV GI and 0.4 to 1.4 log10 for NoV GII. NoVs were not detected in the reuse water produced by MBR/RO system, while sand-anthracite filters resulted in a NoV GI and GII decay of 1.1-1.6 log10 and 0.7-1.6 log10, respectively. A variety of genotypes (GI.2, GI.3a, GI.3b, GI.5, GII.1, GII.4 Sydney 2012, GII.5, GII.6, GII.17) was observed, with a predominance of GI.2 and GII.17 in the different genogroups. These results corroborate with recent data about the entry and dissemination of the emerging genotype GII.P17-GII.17 Kawasaki 2014 in the country, and may indicate a change in the epidemiological patterns of norovirus strains circulation in this region. This is the first large-scale study to evaluate burden and genotypes of noroviruses in WWTPs in Brazil, providing a rapid diagnosis of viruses circulating in the population.

Influence of water quality on diversity and composition of fungal communities in a tropical river.

Freshwater fungi are key decomposers of organic material and play important roles in nutrient cycling, bio-remediation and ecosystem functioning. Although aquatic fungal communities respond to pollution, few studies have quantitatively assessed the effect of freshwater contamination on fungal diversity and composition; and knowledge is scarcer for tropical systems. Here we help fill this knowledge gap by studying a heavily-contaminated South American river spanning a biodiversity hotspot. We collected 30 water samples scattered across a quality gradient over two seasons and analyzed them using Terminal Restriction Fragment Length Polymorphisms (T-RFLP) coupled with 454 Pyrosequencing. Using T-RFLP we identified 451 and 442 Operational Taxonomy Units (OTUs) in the dry and rainy seasons respectively, whereas Pyrosequencing revealed 48,553 OTUs from which 11% were shared between seasons. Although 68% of all identified OTUs and 51% of all identified phyla remained unidentified, dominant fungal phyla included the Ascomycota, Basidiomycota, Chytridiomycota, Glomeromycota, Zygomycota and Neocallimastigomycota, while Calcarisporiella, Didymosphaeria, Mycosphaerella (Ascomycota) and Rhodotorula (Basidiomycota) were the most abundant genera. Fungal diversity was affected by pH and dissolved iron, while community composition was influenced by dissolved oxygen, pH, nitrate, biological oxygen demand, total aluminum, total organic carbon, total iron and seasonality. The presence of potentially pathogenic species was associated with high pH. Furthermore, geographic distance was positively associated with community dissimilarity, suggesting that local conditions allowed divergence among fungal communities. Overall, our findings raise potential concerns for human health and the functioning of tropical river ecosystems and they call for improved water sanitation systems.

Distribution of human fecal marker GB-124 bacteriophages in urban sewage and reclaimed water of São Paulo city, Brazil.

Bacteriophages infecting Bacteroides fragilis GB-124 have been described as potential markers of human fecal contamination in water sources. The aim of this study was to evaluate the occurrence of GB-124 phages in raw sewage, secondary effluents and reclaimed water of the São Paulo city using a low-cost microbial source tracking method. Samples were collected monthly from April 2015 to March 2016 in four municipal wastewater treatment plants that operate with activated sludge processes followed by different tertiary treatments (sand-anthracite filtration, membrane bioreactor/reverse osmosis) and final chlorination. GB-124 phages were detected in 100% of the raw sewage samples, with viral loads varying from 7.5 × 103 to 1.32 × 106 PFU/L. Virus removal efficiency in activated sludge processes ranged from 1.89 to 2.31 log10. Frequencies of phage detection were lower in reclaimed water samples (0-22.2%). The results indicated that GB-124 phage could be a complementary low-cost viral marker for the detection of human fecal pollution in waters impacted with urban sewage in this region. However, the datasets of tertiary effluents resulted in several samples with concentrations below the detection limit (DL ≤1 PFU/mL) suggesting the need to obtain analytical methods with lower DL for greater accuracy of negative results.

Detection and molecular characterization of Cryptosporidium species and Giardia assemblages in two watersheds in the metropolitan region of São Paulo, Brazil.

Cryptosporidium and Giardia are associated with cases of water and foodborne outbreaks in the world. This study included 50 samples of surface raw water collected from two watersheds in the state of São Paulo, Brazil. The isolation of (oo)cysts was performed in accordance with the U.S. Environmental Protection Agency's methods 1623 and genotypic characterization and quantification were carried out by Nested PCR and qPCR assays based on 18S rRNA and gdh genes, respectively. U.S. EPA 1623 method showed the presence of (oo)cysts in 40% ([Formula: see text] = 0.10 oocysts/L) and 100% ([Formula: see text] = 7.6 cysts/L) of samples from São Lourenço River, respectively, and 24% ([Formula: see text] = 0.8 oocysts/L) and 60% ([Formula: see text] = 1.64 cysts/L) of Guarapiranga Reservoir, respectively. The qPCR assay detected C. hominis/parvum in 52% (0.06 to 1.85 oocysts/L) of São Lourenço River and 64% (0.09 to 1.4 oocysts/L) of Guarapiranga Reservoir samples. Presence/absence test for Giardia intestinalis was positive in 92% of São Lourenço River and 8% of Guarapiranga Reservoir samples. The assemblage A was detected in 16% (0.58 to 2.67 cysts/L) in São Lourenço River and no positive samples were obtained for assemblage B in both water bodies. The characterization of anthroponotic species C. parvum/hominis, G. intestinalis, and assemblage A was valuable in the investigation of possible sources of contamination in the watersheds studied confirming the need of expanding environmental monitoring measures for protection of these water sources in our country.

Characterization of Enterococcus species isolated from marine recreational waters by MALDI-TOF MS and Rapid ID API® 20 Strep system.

MALDI-TOF Mass Spectrometry Biotyping has proven to be a reliable method for identifying bacteria at the species level based on the analysis of the ribosomal proteins mass fingerprint. We evaluate the usefulness of this method to identify Enterococcus species isolated from marine recreational water at Brazilian beaches. A total of 127 Enterococcus spp. isolates were identified to species level by bioMérieux's API® 20 Strep and MALDI-TOF systems. The biochemical test identified 117/127 isolates (92%), whereas MALDI identified 100% of the isolates, with an agreement of 63% between the methods. The 16S rRNA gene sequencing of isolates with discrepant results showed that MALDI-TOF and API® correctly identified 74% and 11% of these isolates, respectively. This discrepancy probably relies on the bias of the API® has to identify clinical isolates. MALDI-TOF proved to be a feasible approach for identifying Enterococcus from environmental matrices increasing the rapidness and accuracy of results.

Occurrence of Giardia intestinalis and Cryptosporidium sp. in wastewater samples from São Paulo State, Brazil, and Lima, Peru.

The objectives of the study were to detect and genotype Cryptosporidium spp. and Giardia intestinalis in wastewater samples obtained from five cities with high transit of people in the State of São Paulo, Brazil, and at the entrance of a Wastewater Treatment Plant (WWTP) in Lima, Peru. Samples were collected and concentrated by centrifugation. The genomic DNA was extracted for molecular characterization by nested PCR for Cryptosporidium and double nested PCR for Giardia, followed by sequencing and phylogenetic analysis. G. intestinalis was found in 63.6 % of the samples, and the human assemblages A and B were identified. Cryptosporidium sp. was found in 36.4 % of the samples, and the species were corresponding to Cryptosporidium hominis, Cryptosporidium cuniculus, and Cryptosporidium muris. Results revealed the presence of human pathogenic Cryptosporidium species and G. intestinalis human pathogenic assemblages. Molecular tools highlight the importance to map the genetic diversity of these parasites, as well as to detect their epidemiological circulation pathway in the environment.

Assessing the probability of infection by Salmonella due to sewage sludge use in agriculture under several exposure scenarios for crops and soil ingestion.

A deeper understanding about the risks involved in sewage sludge practice in agriculture is required. The aims of the present study were to determine the annual risk of infection of consuming lettuce, carrots and tomatoes cultivated in soil amended with sewage sludge. The risk to agricultural workers of accidental ingestion of sludge or amended soil was also investigated. A Quantitative Microbial Risk Assessment was conducted based on Salmonella concentrations from five WWTPs were used to estimate the probability of annual infection associated with crops and soil ingestion. The risk of infection was estimated for nine exposure scenarios considering concentration of the pathogen, sewage sludge dilution in soil, variation of Salmonella concentration in soil, soil attachment to crops, seasonal average temperatures, hours of post-harvesting exposure, Salmonella regrowth in lettuce and tomatoes, Salmonella inhibition factor in carrots, crop ingestion and frequency of exposure, sludge/soil ingestion by agricultural workers and frequency of exposure. Annual risks values varied across the scenarios evaluated. Highest values of annual risk were found for scenarios in which the variation in the concentration of Salmonella spp. in both soil and crops (scenario 1) and without variation in the concentration of Salmonella spp. in soil and variation in crops (scenario 3) ranging from 10(-3) to 10(-2) for all groups considered. For agricultural workers, the highest annual risks of infection were found when workers applied sewage sludge to agricultural soils (2.26×10(-2)). Sensitivity analysis suggests that the main drivers for the estimated risks are Salmonella concentration and ingestion rate. These risk values resulted from conservative scenarios since some assumptions were derived from local or general studies. Although these scenarios can be considered conservative, the sensitivity analysis yielded the drivers of the risks, which can be useful for managing risks from the fresh products chain with stakeholders' involvement.

A survey on gram-negative bacteria in saffron finches (Sicalis flaveola) from illegal wildlife tradein Brazil / Pesquisa de bactérias gram-negativas em canários-da-terra (Sicalis flaveola) resgatados do tráfico ilegal de aves no Brasil

Passerines such as canaries or finches are the most unlawfully captured species that are sent to wildlife centers in São Paulo, Brazil. Captured birds may have infection by opportunistic bacteria in stressful situations. This fact becomes relevant when seized passerine are reintroduced. The aim of this study was to evaluate the health state of finches from illegal wildlife trade using microbiological approaches. Microbiological samples were collected by cloacal and tracheal swabs of 100 birds, captured during 2012 and 2013. The results indicate high frequency of gram-negative bacteria in feces and oropharynx, especially from the Enterobacteriaceae family (97.5%). The most frequent genera were Escherichia coli (46.5%) and Klebsiella pneumoniae (10.4%). Enterobacter cloacae, Serratia liquefaciens, Serratia spp. Klebsiella oxytoca and Citrobacter freundii were isolated with lower frequency from asymptomatic birds. The presence of enteropathogenic Escherichia coli (EPEC) and Shiga toxinproducing strain (STEC) confirm the zoonotic risks and public health concern.(AU)

No Estado de São Paulo, Brasil, os pássaros como os canários-da-terra têm sido uma das espécies mais frequentemente resgatadas do tráfico ilegal e enviadas aos centros de vida selvagem. Em situações de estresse estas aves podem ser acometidas por infecções causadas por bactérias oportunistas. Este fato é de grande importância quando é planejada da reintrodução das aves na natureza. O presente trabalho foi delineado para avaliar o estado de saúde de canários-da-terra resgatados do tráfico ilegal. Foram colhidas soabes da traqueia e da cloaca de 100 aves resgatadas durante os anos de 2012 e 2013. Os resultados obtidos revelaram alta frequência de bactérias gram-negativas nas fezes e no orofaringe dos animais, com maior frequência para os membros da família Enterobacteriaceae (97,5%). Os gêneros mais frequentes foram Escherichia coli (46,55) e Klebsiella pneumoniae (10,4%). Outros microorganismos incluindo Enterobacter cloacae, Serratia liquefaciens, Serratia spp, Klebsiella oxytoca e Citrobacter freundii também foram isolados em menor frequencia de aves assintomáticas. A presença de estirpes de Escherichia coli enteropagênicas (EPEC) e as produtoras da toxina de Shiga confirmam o risco de zoonose e a importância para saúde pública deste tipo de ave.(AU)