Search

Summary Objectives Although primarily transmitted by Aedes aegypti mosquitos, Zika virus (ZIKV) can also be transmitted by blood transfusion, due to the fact that some of the infected donors can establish asymptomatic viremia. ZIKV seroprevalence in Brazilian blood donors is unknown. The main reason for this gap in the knowledge originates from the difficulty in evaluating ZIKV humoral immunity due to antigenic cross-reactivity between the different Brazilian flaviviruses and, in particular, dengue virus (DENV). The objective of this study was to evaluate the anti-ZIKV IgG prevalence in blood donors from the Northeast region of the Sao Paulo State, Brazil, which experienced a ZIKV outbreak in 2016. Methods We evaluated the ZIKV seroprevalence using the NS1 anti-ZIKV IgG test (Euroimmun), followed by confirmation of the positive and borderline results using the Plaque Reduction Neutralization Test (PRNT). ZIKV seroprevalence was estimated by testing plasma samples collected in 2015 (before the ZIKV outbreak), 2016 (during the outbreak) and 2017 (after the outbreak). In order to investigate possible antigenic cross – reactivity between ZIKV and DENV we also included samples that were taken well before the ZIKV outbreak, in years 2010 and 2013. Results The results obtained by the Euroimmun anti-ZIKV IgG test demonstrated ZIKV seroreactivity in 2015, 2016, and 2017 with prevalences of 5.3%, 12.8% and 13.2%, respectively. The inclusion of blood donor samples from 2010 to 2013, demonstrated anti-ZIKV IgG reactivity only for 2013 (1.7%). The PRNT testing of the ZIKV positive and borderline ELISA reacting samples generated positive results only for the years of 2016 and 2017 (prevalences of 5.6% and 9.1%) which coincided with the introduction of ZIKV in our region. Conclusions Our results estimate for the first time the ZIKV seroprevalence among Brazilian blood donors from a region with apparently extensive ZIKV circulation and which, at the same time, is highly endemic for DENV. We detected relatively low ZIKV seroprevalence in blood donors from the studied region probably due to the lower intensity of the outbreak compared to other Brazilian locations. Our study adds to the global understanding of ZIKV circulation and the herd immunity of the exposed population.

The intensification of dog, cat and livestock vaccination campaigns significantly reduced rabies cases in humans and domestic animals in Ceará State, Brazil. However, sylvatic animals-bats (order Chiroptera), wild canids, raccoons and non-human primates- remain as reservoirs for the virus. Our hypothesis is that surveillance and monitoring of rabies virus in bats, especially passive surveillance, is of fundamental importance, besides the implementation of health education and strengthening of surveillance actions in humans exposed to aggressions. Thus, we assessed the occurrence of rabies virus in animals focusing on bats, before and after launching of the Sylvatic Rabies Surveillance Program in 2010. Surveillance data from the 184 municipalities of Ceará State were analysed, collected during the periods 2003-2010 (active surveillance) and 2011-2016 (passive surveillance), respectively. A total of 13,543 mammalian samples were received for rabies diagnosis from 2003 to 2016. Of these, 10,960 were from dogs or cats (80.9%), 1,180 from bats (8.7%), 806 from other sylvatic animals (foxes, marmosets, raccoons; 6.0%) and 597 from herbivores (cattle, goats, sheep, equines, pigs; 4.4%). A total of 588 (4.3%) samples were positive for rabies. About 8.4% (99/1,180) of the bat samples were infected with rabies virus, 92 (92.9%) of these were from non-haematophagous bat species and 7 (7.1%) from haematophagous species. The number of bat samples received and infection rates increased considerably, after a shift from active surveillance (9/355 [2.5%] samples positive), to passive surveillance (90/825 [10.9%] samples positive). Surveillance of rabies virus in bats is fundamental for human and domestic animal health in Ceará State. Bats have to be considered as targets in surveillance and control programmes. Virus lineages should be characterized to increase knowledge on transmission dynamics of sylvatic rabies virus to domestic animals and the human population, and to provide additional evidence for planning and implementation of improved control measures.

Objectives: Defense against respiratory viruses depends on an immune response present in the mucosa, as saliva IgA secretes antibodies. During the pandemic, such as influenza or SARS-CoV-2, most infected patients are asymptomatic but retain specific antibodies post-infection. The authors evaluated IgG and IgA antibodies against SARS-CoV-2 and influenza in the saliva of asymptomatic volunteers, validated with controls or vaccinated individuals. Methods: The authors detected specific antibodies by validated conventional ELISA using natural SARS-CoV-2 antigens from infected Vero cells or capture-ELISA for influenza using natural antigens of the influenza vaccine. Results: Saliva from influenza-vaccinated individuals had more IgA than paired serum, contrary to the findings for specific IgG. In COVID-19-vaccinated samples, specific IgA in saliva increased after vaccination, but IgG levels were high after the first dose. In saliva from the asymptomatic population (226), anti-Influenza IgG was found in 57.5% (130) of samples, higher than IgA, found in 35% (79) of samples. IgA results were similar for SARS-CoV-2, with IgA present in 30% (68) of samples, while IgG was less present, in 44.2% (100) of samples. The proportion of influenza IgG responders was higher than that for SARS-CoV-2 IgG, but both populations presented similar proportions of IgA responders, possibly due to variable memory B cell survival. For both viruses, the authors found an important proportion (> 10%) of IgA+IgG- samples, suggesting the occurrence of humoral immunity directed to the mucosa. Conclusion: Specific antibodies for respiratory viruses in saliva are found in either infection or vaccination and are a convenient and sensitive diagnostic tool for host immune response.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the agent of coronavirus disease 2019 (COVID-19), is responsible for the worst pandemic of the 21st century. Like all human coronaviruses, SARS-CoV-2 originated in a wildlife reservoir, most likely from bats. As SARS-CoV-2 has spread across the globe in humans, it has spilled over to infect a variety of non-human animal species in domestic, farm, and zoo settings. Additionally, a broad range of species, including one neotropical monkey, have proven to be susceptible to experimental infection with SARS-CoV-2. Together, these findings raise the specter of establishment of novel enzootic cycles of SARS-CoV-2. To assess the potential exposure of free-living non-human primates to SARS-CoV-2, we sampled 60 neotropical monkeys living in proximity to Manaus and São José do Rio Preto, two hotspots for COVID-19 in Brazil. Our molecular and serological tests detected no evidence of SAR-CoV-2 infection among these populations. While this result is reassuring, sustained surveillance efforts of wildlife living in close association with human populations is warranted, given the stochastic nature of spillover events and the enormous implications of SARS-CoV-2 spillover for human health.

Zika virus (ZIKV) is a mosquito-borne viral disease associated with fetal microcephaly and other central nervous system (CNS) symptomatology. It was first identified in a Rhesus macaque in Uganda in 1947 and later in humans (Zika fever). In 2015, ZIKV was notified in Northeast Brazil where it was associated with CNS alterations and with rapid epidemic spread. Considering that ZIKV infects Old World monkeys, the aim of this study was to follow its potential in neotropical primates. Here, we show the detection of ZIKV in marmosets and capuchin monkeys captured in Ceara state, Northeast Brazil. Nine (9/132) samples were positive by quantitative RT-PCR assay. Neutralizing antibodies in primates for ZIKV were also detected by PRNT. The ZIKV-positive samples were obtained from peridomestic animals captured in proximity to humans in areas with reports of ZIKV-associated microcephaly cases during the epidemic period. These results reiterate the molecular evidence of ZIKV infection in neotropical primates, and the temporal detection suggests that detection in primates occurred during the epidemic period in humans. However, a continuous surveillance is necessary to exclude the possibility of virus circulation and transmission in wild environments.

Background Current evidence regarding COVID‐19 convalescent plasma (CCP) transfusion practices is limited and heterogeneous. We aimed to determine the impact of the use of CCP transfusion in patients with previous circulating neutralizing antibodies (nAbs) in COVID‐19. Methods Prospective cohort including 102 patients with COVID‐19 transfused with ABO compatible CCP on days 0–2 after enrollment. Clinical status of patients was assessed using the adapted World Health Organization (WHO) ordinal scale on days 0, 5, and 14. The nAbs titration was performed using the cytopathic effect‐based virus neutralization test with SARS‐CoV‐2 (GenBank MT126808.1). The primary outcome was clinical improvement on day 14, defined as a reduction of at least two points on the adapted WHO ordinal scale. Secondary outcomes were the number of intensive care unit (ICU)‐free days and the number of invasive mechanical ventilation‐free days. Results Both nAbs of CCP units transfused (p

Background: The risk of contamination and dissemination by SARS-CoV-2 has a strong link with nasal, oral and pharyngeal cavities. Recently, our research group observed the promising performance of an anionic phthalocyanine derivative (APD) used in a mouthwash protocol without photoexcitation; this protocol improved the general clinical condition of patients infected with SARS-CoV-2. Methods: The present two-arm study evaluated in vitro the antiviral activity and cytotoxicity of APD. Additionally, a triple-blind randomized controlled trial was conducted with 41 hospitalized patients who tested positive for COVID-19. All the included patients received World Health Organization standard care hospital treatment (non-intensive care) plus active mouthwash (experimental group AM/n=20) or nonactive mouthwash (control group NAM/n=21). The adjunct mouthwash intervention protocol used in both groups consisted one-minute gargling/rinsing / 5 times/day until hospital discharge. Groups were compared considering age, number of comorbidities, duration of symptoms prior admission and length of hospital stay (LOS). The associations between group and sex, age range, presence of comorbidities, admission to Intensive care unit (ICU) and death were also evaluated. Results: The in vitro evaluation demonstrated that APD compound was highly effective for reduction of SARS-CoV-2 viral load in the 1.0 mg/mL (99.96%) to 0.125 mg/mL (92.65%) range without causing cytotoxicity. Regarding the clinical trial, the median LOS of the AM group was significantly shortened (4 days) compared with that of the NAM group (7 days) (p=0.0314). Additionally, gargling/rinsing with APD was very helpful in reducing the severity of symptoms (no ICU care was needed) compared to not gargling/rinsing with APD (28.6% of the patients in the NAM group needed ICU care, and 50% of this ICU subgroup passed way, p=0.0207). Conclusions: This study indicated that the mechanical action of the protocol involving mouthwash containing a compound with antiviral effects against SARS-CoV-2 may reduce the symptoms of the patients and the spread of infection. The use of APD in a mouthwash as an adjuvant the hospital COVID-19 treatment presented no contraindication and reduced the hospital stay period. Trial Registration: The clinical study was registered at REBEC - Brazilian Clinical Trial Register (RBR-58ftdj) in 10/28/2020.

BACKGROUND: Blood groups and anti-A isohemagglutinin may be involved in susceptibility to SARS-CoV-2 infection. MATERIALS AND METHODS: We retrospectively studied 268 COVID-19 convalescent plasma donors and 162 COVID-19 inpatients (total 430 subjects, confirmed by RT-PCR) and 2,212 healthy volunteer first-time blood donors as a control group. These were further divided into two groups: those with anti-A (blood types O and B) and those without it (types A and AB). Titres of nucleoproteins, and neutralizing SARS-CoV-2 antibody were measured in the convalescent plasma donors and inpatients. Multivariate logistic regression and non-parametric tests were applied. RESULTS: Persons having types O or B showed less infection prevalence than those of types A or AB (OR = 0·62, 95% CI 0·50-0·78; P

Background The efficacy of convalescent plasma (CP), an alternative for the treatment of COVID‐19, depends on high titers of neutralizing antibodies (nAbs), but assays for quantifying nAbs are not widely available. Our goal was to develop a strategy to predict high titers of nAbs based on the results of anti‐SARS‐CoV‐2 immunoassays and the clinical characteristics of CP donors. Study Design and Methods A total of 214 CP donors were enrolled and tested for the presence of anti‐SARS‐CoV‐2 antibodies (IgG) using two commercial immunoassays: EUROIMMUN (ELISA) and Abbott (Chemiluminescence). Quantification of nAbs was performed using the Cytopathic Effect‐based Virus Neutralization test. Three criteria for identifying donors with nAbs ≥ 1:160 were tested: – C1: Curve ROC; − C2: Conditional decision tree considering only the IA results and – C3: Conditional decision tree including both the IA results and the clinical variables. Results The performance of the immunoassays was similar referring to both S/CO and predictive value for identifying nAbs titers ≥1:160. Regarding the studied criteria for identifying CP donors with high nAbs titers: (a) C1 showed 76.1% accuracy if S/CO = 4.65, (b) C2 presented 76.1% accuracy if S/CO ≥4.57 and (c) C3 had 71.6% accuracy if S/CO was ≥4.57 or if S/CO was between 2.68‐4.57 and the last COVID‐19‐related symptoms were recent (within 19 days). Conclusion SARS‐CoV‐2 IgG immunoassays (S/CO) can be used to predict high anti‐SARS‐CoV‐2 nAbs titers. This study has proposed different criteria for identifying donors with ≥1:160 nAbs titers, all with high efficacy.

This study aims to describe the sociodemographic determinants associated with exposure to Zika Virus (ZIKV) in pregnant women during the 2015–2016 epidemic in Salvador, Brazil. Methods We recruited women who gave birth between October 2015 and January 2016 to a cross-sectional study at a referral maternity hospital in Salvador, Brazil. We collected information on their demographic, socioeconomic, and clinical characteristics, and evaluated their ZIKV exposure using a plaque reduction neutralization test. Logistic regression was then used to assess the relationship between these social determinants and ZIKV exposure status. Results We included 469 pregnant women, of whom 61% had a positive ZIKV result. Multivariate analysis found that lower education (adjusted Prevalence Rate [aPR] 1.21; 95%CI 1.04–1.35) and food insecurity (aPR 1.17; 95%CI 1.01–1.30) were positively associated with ZIKV exposure. Additionally, age was negatively associated with the infection risk (aPR 0.99; 95%CI 0.97–0.998). Conclusion Eve after controlling for age, differences in key social determinants, as education and food security, were associated with the risk of ZIKV infection among pregnant women in Brazil. Our findings elucidate risk factors that can be targeted by future interventions to reduce the impact of ZIKV infection in this vulnerable population., Author summary The Zika virus (ZIKV) epidemic in Brazil has intensified global concern about congenital defects associated with intrauterine exposure. Social determinants are factors that reinforce and contribute to the transmission and spread of ZIKV as well as other arboviruses like Dengue. We performed a cross-sectional study to describe the prevalence of ZIKV and the contribution of social determinants to transmission among pregnant women during the 2015–2016 ZIKV epidemic in Salvador, Brazil. We found that 61% of pregnant women were ZIKV seropositive. We also found that lower education level, food insecurity and lower maternal age were associated with higher ZIKV infection risk. These findings contribute to understanding the role of social determinants in ZIKV transmission, providing key social factors that can be combined with pre-existing tactics (vector control and environmental improvement) to create policies and interventions which reduce social inequalities and risk of infection in vulnerable populations like pregnant women.

An unprecedented global health crisis has been caused by a new virus called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We performed experiments to test if a hypertonic saline solution was capable of inhibiting virus replication. Our data show that 1.2% NaCl inhibited virus replication by 90%, achieving 100% of inhibition at 1.5% in the nonhuman primate kidney cell line Vero, and 1.1% of NaCl was sufficient to inhibit the virus replication by 88% in human epithelial lung cell line Calu-3. Furthermore, our results indicate that the inhibition is due to an intracellular mechanism and not to the dissociation of the spike SARS-CoV-2 protein and its human receptor. NaCl depolarizes the plasma membrane causing a low energy state (high ADP/ATP concentration ratio) without impairing mitochondrial function, supposedly associated with the inhibition of the SARS-CoV-2 life cycle. Membrane depolarization and intracellular energy deprivation are possible mechanisms by which the hypertonic saline solution efficiently prevents virus replication in vitro assays.

INTRODUCTIONCOVID-19 convalescent plasma (CCP) transfusion has emerged in the past months as an alternative approach to treat pneumonia cases of SARS-CoV-2. Current evidence regarding characteristics of the plasma product, the titer of neutralizing antibodies (nAbs) in the transfused units, time to onset of intervention, and impact of nAbs produced by the patient are limited and heterogeneous.MATERIAL AND METHODSWe describe the preliminary results of 104 patients with severe pneumonia due to SARS-CoV-2 transfused with CCP at three medical centers in Brazil. All enrolled patients were transfused with doses between 200 mL through 600mL of ABO compatible CCP on days 0-2 after enrolment. Clinical parameters were monitored and nAbs titration was performed using the cytopathic effect-based virus neutralization test with SARS-CoV-2 (GenBankMT126808.1).RESULTSForty-one patients achieved clinical improvement on day 14, and multivariable logistic regression showed that nAbs T (from CCP units transfused) (p= 0.001), nAbs P0 (on day of enrolment) (p=0.009) and use of other supportive therapies (pDISCUSSION/CONCLUSIONDespite the study limitations, our data have shown an association between patients’ previously acquired nAbs and clinical outcomes. The potential value of timely administration of CCP transfusion before day 10 of disease onset was demonstrated and nAbsP0, but not nAbsT, were associated with ICU LOS, and duration of mechanical ventilation on the improvement of clinical outcomes was also demonstrated. In conclusion, we consider these data are useful parameters to guide future CPP transfusion strategies to COVID-19.

IntroductionA cohort of COVID-19 convalescent volunteers allowed the study of neutralizing (nAb) and ligand antibodies kinetics by providing sequential samples during a median of 100 days after onset of disease.Material and MethodsA cohort of previously RT-PCR+ve (detected by nasopharyngeal swab during the acute phase), male convalescent patients, all with mild symptoms, were enrolled on serial blood sample collection for evaluation of longitudinal nAb titers and anti-nucleocapsid (NP) antibodies (IgM, IgG and IgA). Nabs were detected by a cytopathic effect-based virus neutralization test (CPE-based VNT), carried out with SARS-CoV-2 (GenBank: MT350282)ResultsA total of 78 male volunteers provided 316 samples, spanning a total of 4820 days of study. Although only 25% of donors kept nAb titers ≥160, after a median of 100 days after the onset of disease, there was a high probability of sustaining nAB titers ≥160 in volunteers whose initial nAb titer was ≥1280, weight ≥ 90kg or BMI classified as overweight or obese, evidenced by Kaplan-Meier estimates and Cox hazard regression. There was no correlation between ABO group, ABO antibody titers and persistent high nAb titers. High IgG anti-NP (S/CO ≥5.0) is a good surrogate for detecting nAB ≥160, defined by ROC curve (sensitivity = 90.5%; CI95% 84.5-94.7%)ConclusionSelection of CCP donors for multiple collections based on initial high nAb titers (≥1280) or overweight/obese (BMI) provides a simple strategy to achieve higher quality in CCP programs. High IgG anti-NP levels can also be used as surrogate markers for high nAb screening.

Background: Passive antibody therapy with convalescent plasma (CP) represents a promising alternative for the treatment of SARS CoV 2 infection. The efficacy of CP therapy has been associated with high titers of neutralizing antibodies (nAbs) in the plasma of recovered patients, but the assays for quantifying nAbs are not widely available. Our goal was to develop a strategy to predict high titers of nAbs based on the results of anti-SARS CoV 2 immunoassays and the clinical characteristics of the CP potential donors. Methods: Two hundred and fourteen CP donors were enrolled and tested for the presence of anti-SARS CoV 2 antibodies using two commercial immunoassays (IA): Anti SARS CoV 2 ELISA IgG EUROIMMUN and Anti SARS CoV 2 Chemiluminescence IgG Abbott. In parallel, quantification of neutralizing antibodies (nAbs) was performed using the Cytopathic effect-based virus neutralization test (CPE VNT). Three criteria for identifying donors with high titers of nAbs (more than 160) were tested: Criterion1: Curve ROC Method; Criterion 2: Conditional decision tree considering only the results from the IA and Criterion 3: Conditional decision tree including both the IA results and the clinical variables. Results: The performance of Abbott and EUROIMMUN immunoassays was similar referring to both S/CO and predictive value for identifying nAbs titers more than 1:160. Regarding the three studied criteria for identifying CP donors with high nAbs titers (more than 1:160): 1) Criterion 1 showed 76.1% accuracy when the S/CO cut-off of 4.65 was used, 2) Criterion 2 presented 76.1% accuracy if the S/CO more than 4.57 was applied and 3) Criterion 3 had 71.6% accuracy if either S/CO more than 4.57 or S/CO between 2.68 and 4.57 and the last COVID-19-related symptoms occurred less than 19 days from donor recruiting were used. Conclusion: The results of SARS-CoV-2 immunoassays (S/CO) can be used to predict high nAbs titers of potential CP donors. This study has proposed three different criteria for identifying donors with more than 160 nAbs titer based on either solely S/CO results or S/CO together with clinical variables, all with high efficacy and accuracy.

Although first considered a benign infection, recent studies have disclosed severe and potentially lethal inflammatory manifestations of COVID-19 in children. We report the case of a 4-year-old child with a post-infectious multisystem inflammatory syndrome associated with COVID-19, with a Kawasaki-like shock and prominent neurologic features, for whom a cytokine storm and reduced brain-derived neurotrophic factor were well documented.

In March 2020, WHO declared a pandemic state due to SARS-CoV-2 having spread. TaqMan-based real-time RT-qPCR is currently the gold standard for COVID-19 diagnosis. However, it is a high-cost assay, inaccessible for the majority of laboratories around the world, making it difficult to diagnose on a large scale. The objective of this study was to standardize lower cost molecular methods for SARS-CoV-2 identification. E gene primers previously determined for TaqMan assays by Colman et al. (2020) were adapted in SYBR Green assay and RT-PCR conventional. The cross-reactivity test was performed with 17 positive samples for other respiratory viruses, and the sensibility test was performed with 8 dilutions (10 based) of SARS-CoV-2 isolated and 63 SARS-CoV-2-positive samples. The SYBR Green assays and conventional RT-PCR have not shown amplification of the 17 respiratory samples positives for other viruses. The SYBR Green-based assay was able to detect all 8 dilutions of the isolate. The conventional PCR detected until 107 dilution, both assays detected the majority of the 63 samples, 98.42% of positivity in SYBR Green, and 93% in conventional PCR. The average Ct variation between SYBR Green and TaqMan was 1.92 and the highest Ct detected by conventional PCR was 35.98. Both of the proposed assays are less sensitive than the current gold standard; however, our data shows a low sensibility variation, suggesting that these methods could be used by laboratories as a lower cost molecular method for SARS-CoV-2 diagnosis. Electronic supplementary material The online version of this article (10.1007/s42770-020-00347-5) contains supplementary material, which is available to authorized users.

Background Coronavirus disease 2019 (COVID‐19) convalescent plasma (CCP) collection began in two Brazilian hospitals for treatment of severe/critical patients. Methods and Materials Mild/moderate COVID‐19 convalescents were selected as CCP donors after reverse transcription polymerase chain reaction (RT‐PCR) confirmed severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection and absence of symptoms for ≥14 days plus (a) age (18‐60 years), body weight greater than 55 kg; (b) immunohematological studies; (c) no infectious markers of hepatitis B virus, hepatitis C virus, human immunodeficiency virus, human T‐lymphotropic virus‐1/2, Chagas and syphilis infection; (d) no HLA antibodies (multiparous); (e) second RT‐PCR (nasopharyngeal swab and/or blood) negativity; (f) virus neutralization test (cytopathic effect–based virus neutralization test neutralizing antibody) and anti–nucleocapsid protein SARS‐CoV‐2 IgM, IgG, and IgA enzyme‐linked immunosorbent assays. Results Among 271 donors (41 females, 230 males), 250 presented with neutralizing antibodies. Final RT‐PCR was negative on swab (77.0%) or blood (88.4%; P = .46). Final definition of RT‐PCR was only defined at more than 28 days after full recovery in 59 of 174 (33.9%) RT‐PCR –ve, and 25/69 RT‐PCR +ve (36.2%; 13 between 35 and 48 days). Neutralizing antibody titers of 160 or greater were found in 63.6%. Correlation between IgG signal/cutoff of 5.0 or greater and neutralizing antibody of 160 or greater was 82.4%. Combination of final RT‐PCR –ve with neutralizing antibody ≥160 was 41.3% (112/271). Serial plasma collection showed decline in neutralizing antibody titers and IgA levels (P

Objective To describe the neurological and neurodevelopmental outcomes of children with Congenital Zika Syndrome (CZS) associated microcephaly beyond 2 years of age. Method We followed children with CZS-associated microcephaly in an outpatient clinic in Salvador, Brazil. Neurological and neurodevelopmental assessments were performed using the Hammersmith Infant Neurological Examination (HINE) and Bayley Scales of Infant and Toddler Neurodevelopment (Bayley-III) respectively. Results Of the 42 children included, 19 were male (45.2%); median (interquartile range) age at neurological evaluation was 28 (25–32) months, and 36 (85.7%) had severe microcephaly. HINE and Bayley-III results were completed for 35/42 (83.3%) and 33/42 (78.5%) children respectively. Bayley-III identified a severe developmental delay in 32/33 (97.0%) children while 1/33 (3.0%) had only a mild delay. In the multivariable analysis, we found that Bayley-III and HINE scores were correlated. Better HINE scores were associated with higher Bayley-III cognitive raw scores (β = 0.29; CI 95% = 0.02–0.57) and motor raw scores (β = 0.43; CI 95% = 0.04–0.82) after adjusting for head circumference, prematurity, and age at neurodevelopmental evaluation. Furthermore, we found that greater head circumference at follow up was associated with higher cognitive (β = 1.27; CI 95% = 0.01–2.53) and motor raw scores (β = 2.03; CI 95% = 0.25–3.81). Conclusion Children with CZS-associated microcephaly demonstrate severe neurodevelopmental delays and slower growth rates than their peers over time. Still, they have remarkably heterogeneous neurodevelopmental profiles according to neurological exam scores which correlate with their long-term outcomes. We found that HINE scores effectively captured the heterogeneity of neurological capabilities among these children and could be predictive of cognitive and motor development progress.

Paraguay has registered no human cases of rabies since 2004, and the last case in dogs, reported in 2009, was due to a variant maintained in the common vampire bat "Desmodus rotundus". In 2014, a dog was diagnosed as positive for rabies with aggression towards a boy and all required measures of control were successfully adopted. Epidemiological investigation revealed that the dog was not vaccinated and had been attacked by a crab-eating fox, "zorro" (Cerdocyon thous). The sample was diagnosed by the Official Veterinary Service of the Country and sent to the Center on Rabies Research from the University of São Paulo, Brazil, for antigenic and genetic characterization. A second sample from a dog positive for rabies in the same region in 2015 and 11 samples from a rabies outbreak from Asuncion in 1996 were also characterized. The antigenic profile of the samples, AgV2, was compatible with one of the variants maintained by dogs in Latin America. In genetic characterization, the samples segregated in the canine (domestic and wild species)-related group in an independent subgroup that also included samples from Argentina. These results and the epidemiology of the case indicate that even with the control of rabies in domestic animals, the virus can still circulate in wildlife and may be transmitted to domestic animals and humans, demonstrating the importance of continuous and improved surveillance and control of rabies, including in wild species, to prevent outbreaks in controlled areas.

Yellow fever virus RNA is usually detected in blood of infected humans. We detected virus RNA in urine and semen samples from a convalescent patient. A complete virus genome was sequenced for an isolate from a urine sample. This virus had a South American I genotype and unique synapomorphic changes.

The World Health Organization (WHO), reports that rabies causes tens of thousands of deaths every year killing humans, non-human primates and other animals. Rabies continues to be a public health issue, despite the existence of effective vaccines. The dogs remain the primary reservoir and transmitter of rabies to humans globally. In the Americas, bats are regarded as the second most common source of rabies virus to humans. The vampire bat Desmodus rotundus has been identified as a natural reservoir of rabies virus (RABV) in this region. The complete genome of the RABV variant maintained by populations of vampire bats D. rotundus has rarely been reported. In this study, we sequenced and analyzed the genome of a RABV variant detected in D. rotundus. The sample, collected from an endemic area in São Paulo State, was phylogenetically compared with the genome of the standard sample for species Rabies virus as well as other samples belonging to terrestrial and bat-associated cycles of rabies transmission, available in GenBank. Distinct patterns linked to the genetic lineage were identified. These data can aid in the understanding of the molecular epidemiology of this virus and the epidemiological importance of this species in the transmission of the RABV.

Made available in DSpace on 2015-11-03T15:30:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-11-01. Added 1 bitstream(s) on 2015-11-04T10:14:35Z : No. of bitstreams: 1 S0037-86822014000600709.pdf: 936273 bytes, checksum: 2cd1319e1d83075796ede708366c71b5 (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Introduction: Rabies is an important zoonosis that occurs in mammals, with bats acting as Lyssavirus reservoirs in urban, rural and natural areas. Rabies cases in bats have been recorded primarily in urban areas in Northwestern State of Sao Paulo since 1998. This study investigated the circulation of rabies virus by seeking to identify the virus in the brain in several species of bats in this region and by measuring rabies-virus neutralizing antibody levels in the hematophagous bat Desmodus rotundus. Methods: From 2008 to 2012, 1,490 bat brain samples were sent to the Universidade Estadual Paulista (UNESP) Rabies Laboratory in Aracatuba, and 125 serum samples from vampire bats that were captured in this geographical region were analyzed. Results: Rabies virus was detected in the brains of 26 (2%) of 1,314 non-hematophagous bats using the fluorescent antibody test (FAT) and the mouse inoculation test (MIT). None of the 176 hematophagous bat samples were positive for rabies virus when a virus detection test was utilized. Out of 125 vampire bat serum samples, 9 (7%) had levels of rabies virus neutralization antibodies (RVNAs) that were higher than 0.5IU/mL; 65% (81/125) had titers between 0.10IU/mL and 0.5IU/mL; and 28% (35/125) were negative for RVNAs using the simplified fluorescent inhibition microtest (SFIMT) in BHK21 cells. The observed positivity rate (1.7%) was higher than the average positivity rate of 1.3% that was previously found in this region. Conclusions: The high percentage of vampire bats with neutralizing antibodies suggests that recent rabies virus exposure has occurred, indicating the necessity of surveillance measures in nearby regions that are at risk to avoid diffusion of the rabies virus and possible rabies occurrences. Universidade Estadual Paulista (UNESP), Faculdade de Medicina Veterinária de Araçatuba, Araçatuba, SP Polo Regional de Desenvolvimento Tecnológico dos Agronegócios da Alta Sorocabana, Agência Paulista de Tecnologia Agropecuária, Presidente Prudente, SP Curso de Farmácia e Bioquímica, Universidade Paulista, Araçatuba, SP Núcleo de Pesquisas em Raiva, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP Universidade Estadual Paulista (UNESP), Faculdade de Medicina Veterinária de Araçatuba, Araçatuba, SP CNPq: 578281/2008-2 FAPESP: 2008/00976-0

Samples from sera and oral swabs from fifteen marmosets (Callithrix jacchus) and nine capuchin-monkeys (Sapajus libidinosus) captured in Ceara State in Brazil were tested for Zika virus. Samples were positive by Real time PCR and sequencing of the amplified product from a capuchin monkey showed 100% similarity to other ZIKV from South America. This is the first report on ZIKV detection among Neotropical primates.

INTRODUCTION: Rabies is an important zoonosis that causes thousands of deaths worldwide each year. Although the terrestrial cycle, mainly transmitted by dogs, is controlled in Brazil, the aerial cycle remains a serious public health issue, besides the economic problem. In the aerial cycle, the haematophagous bat Desmodus rotundus is the main source of infection, where several different species of non-haematophagous bats can be infected and can transmit the virus. METHODS: The aim of this work was to study the epidemiological pattern of rabies using antigenic characterization with monoclonal antibodies and genetic characterization by reverse-transcriptase polymerase chain reaction followed by sequencing and phylogenetic analysis of non-haematophagous bats' and herbivorous animals' central nervous system samples from the western region of the State of São Paulo, Brazil. RESULTS: From 27 samples, 3 antigenic variants were identified: AgV-3, AgV-4, and AgV-6; and from 29 samples, 5 different clusters were identified, all belonging to the rabies virus species. CONCLUSIONS: Although only non-haematophagous bats were evaluated in the studied region, the majority of samples were from antigenic and genetic variants related to haematophagous bats Desmodus rotundus. Samples from the same antigenic variant were segregated in more than one genetic cluster. This study demonstrated the diversity of rabies virus genetic lineages presented and circulating in non-haematophagous bats in the studied region.

This epidemiological study was conducted using antigenic and genetic characterisation of rabies virus isolates obtained from different animal species in the southeast of Brazil from 1993 to 2007. An alteration in the epidemiological profile was observed. One hundred two samples were tested using a panel of eight monoclonal antibodies, and 94 were genetically characterised by sequencing the nucleoprotein gene. From 1993 to 1997, antigenic variant 2 (AgV-2), related to a rabies virus maintained in dog populations, was responsible for rabies cases in dogs, cats, cattle and horses. Antigenic variant 3 (AgV-3), associated with Desmodus rotundus, was detected in a few cattle samples from rural areas. From 1998 to 2007, rabies virus was detected in bats and urban pets, and four distinct variants were identified. A nucleotide similarity analysis resulted in two primary groups comprising the dog and bat antigenic variants and showing the distinct endemic cycles maintained in the different animal species in this region.

Rabies causes thousands of human and animal deaths worldwide each year. The emergent importance of rabies in wild animals demonstrates the necessity of epidemiologic studies of infection in these species toward the development of better strategies for prevention and control of rabies. We analyzed the circulation of rabies virus among wildlife species from a native rainforest in São Paulo State, Brazil. We used the rapid fluorescent focus inhibition test (RFFIT) to test for rabies virus-neutralizing antibodies in 139 captured terrestrial mammals and the fluorescent antibody test (FAT), mouse inoculation test (MIT), and reverse-transcriptase (RT)-PCR to test for virus in samples from the central nervous system of 53 animals found dead. The percentage of samples positive by RFFIT was 10.8%. All samples tested by FAT, MIT, and RT-PCR were negative. Research should be continued to obtain a better understanding of the role of wildlife in the circulation and transmission of rabies virus.

A crescente importância do ciclo silvestre da raiva envolvendo morcegos e mamíferos terrestres demonstra a importância do estudo da epidemiologia do vírus da raiva nessas espécies. Foram estudadas amostras provenientes de diversas espécies de animais silvestres terrestres procedentes de uma área de Mata Atlântica nativa no litoral Norte do Estado de São Paulo. Este estudo pesquisou a presença de anticorpos contra o vírus da raiva por meio dos Testes de RFFIT, SFIMT e ELISA em amostras de soro de animais capturados em uma área desmatada, transpostos e monitorados em uma área de soltura. E ainda a pesquisa do vírus da raiva por meio das técnicas de IFD, IC e RT-PCR em amostras de sistema nervoso central dos animais encontrados mortos na região. A positividade observada foi de 10,8% na RFFIT, 34,0% no SFIMT e 1,13% no ELISA. Todos as amostras submetidas as técnicas de IFD, IC e RT-PCR apresentaram resultados negativos. Os resultados observados demonstram circulação do vírus entre as espécies silvestres da área. The emergent importance of rabies in terrestrial wild animals demonstrates the importance of epidemiological studies regarding rabies virus in those animal species. Samples from several wild species from a native Rainforest area in the North coast of São Paulo State Brazil were studied. The aim of the present study was the research of anti-rabies antibodies in those animals using the RFFIT, SFIMT and ELISA techniques in animals captured and monitorated in a release área. And also by rabies virus detection using FAT, MIT and RT-PCR techniques in central nervous system samples from animals found dead in the same area. The observed positivity was 10,8% on RFFIT, 34,0% on SFIMT e 1,13% on ELISA. All samples tested by the FAT, MIT and RT-PCR techniques presented negative results. The observed results are an evidence of rabies virus circulation between wild animal species in the studied area.

Full-length genome sequencing of the rabies virus is not a routine laboratory procedure. To understand fully the epidemiology, genetic variation and evolution of the rabies virus, full-length viral genomes need to be obtained. For rabies virus studies, cDNA synthesis is usually performed using nonspecific oligonucleotides followed by cloning. When specific primers are used, the cDNA obtained is only partial and is limited to the coding regions. Therefore, the development of methods for synthesizing long cDNA using rabies virus-specific primers is of fundamental importance. A new protocol for the synthesis of long cDNA and the development of 19 new primers are described in this study. This procedure allowed the efficient amplification of the full-length genome of the rabies virus variant maintained by hematophagous bat (Desmodus rotundus) populations following the synthesis of a complete long cDNA. Partial sequencing of the rabies virus genome was performed to confirm rabies-specific PCR amplification. Because degenerate primers were employed, this technique can be adapted easily to other variants. Importantly, this new method is faster and less expensive than cloning methods.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:23:43Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:30:11Z : No. of bitstreams: 1 2-s2.0-70449500458.pdf: 342733 bytes, checksum: d64bfdb9dd2616b8330628b7753f002a (MD5) Made available in DSpace on 2014-05-27T11:23:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-12-01 Background. The use of methods, both sensitive and specific, for rabies diagnosis are important tools for the control and prophylaxis of the disease. Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) has been used in rabies diagnosis with good results, even in decomposed materials. Additionally, molecular techniques have been used for epidemiological studies and to gain a better knowledge of viral epidemiology. Findings. The aim of this work was to evaluate the RT-PCR and hnRT-PCR for rabies virus detection in original tissues stored at -20°C for different periods considering their use for rabies virus detection in stored and decomposed samples. RT-PCR and hnRT-PCR were evaluated in 151 brain samples from different animal species, thawed and left at room temperature for 72 hours for decomposition. The RT-PCR and hnRT-PCR results were compared with previous results from Direct Fluorescent Antibody Test and Mouse Inoculation Test. From the 50 positive fresh samples, 26 (52%) were positive for RT-PCR and 45 (90%) for hnRT-PCR. From the 48 positive decomposed samples, 17 (34, 3%) were positive for RT-PCR and 36 (75%) for hnRT-PCR. No false-positives results were found in the negatives samples evaluated to the molecular techniques. Conclusion. These results show that the hnRT-PCR was more sensitive than RT-PCR, and both techniques presented lower sensibility in decomposed samples. The hnRT-PCR demonstrated efficacy in rabies virus detection in stored and decomposed materials suggesting it's application for rabies virus retrospective epidemiological studies. © 2008 Arajo et al; licensee BioMed Central Ltd. UNESP School of Veterinary and Animal Science Department of Veterinary Hygiene and Public Health, Botucatu, SP Center of Zoonosis Control CCZ, São Paulo, SP UNESP School of Veterinary and Animal Science Department of Veterinary Hygiene and Public Health, Botucatu, SP

O desmatamento e as mudanças de ambiente causadas pelo homem levam a drásticas alterações dos padrões de migração das aves. A contínua perda e fragmentação do habitat ao longo dos corredores das grandes rotas migratórias criam gargalos, resultando em superpopulação e aglomerados de espécies contribuem para aquisição e carreamento de diferentes patógenos ao longo destas rotas, incluindo os coronavirus (CoVs). Maiores vírus de RNA conhecidos, os CoVs possuem grande capacidade de se adaptar a novos hospedeiros e nichos ecológicos, sendo a emergência dos vírus causadores da SARS e da MERS, a partir de vírus de morcegos, os exemplos mais notáveis. Apesar de sua relevância e da complexidade destes agentes, são escassos os estudos de detecção e caracterização genética destes vírus, em especial em aves selvagens. Desta forma este trabalho visa detectar, analisar e discutir a diversidade destes agentes em aves nos locais de parada e invernada de aves migratórias. Para tal, 852 amostras aviárias provenientes de diferentes regiões do Brasil foram submetidas a extração de RNA total seguida das reações de Transcrição Reversa (RT), Reação em Cadeia da Polimerase (PCR) e Nested-PCR, com primers que têm como alvo uma região do ORF 1ab codificante do gene da RpRd viral, e os resultados foram visualizados através de eletroforese em gel de agarose. Deste total, 15 apresentaram resultados positivos para CoVs sendo que em 5 delas foi possível a confirmação por sequenciamento pelo método de Sanger. Adicionalmente, outras 6 amostras de CoVs de um estudo anterior*, foram analisadas visando dar continuidade nos estudos de caracterização viral. Na análise filogenética das sequências da região da ORF 1ab destas 11 amostras, 3 delas se agruparam com o gênero dos Deltacoronavírus. Este grupo tem especial importância por ter um reconhecido potencial de transmissão e emergência de aves para mamíferos, a exemplo do PorCoV HKU15, associado a surtos fatais em suínos. As 8 demais amostras foram agrupadas com o gênero dos Gammacoronavírus, ao qual pertence o vírus causador da doença da Bronquite Infecciosa em galinhas, o mais conhecido CoV aviário, causador de grandes perdas econômicas, que se desenvolve no trato respiratório, reprodutor, gastrointestinal e rins. Com a finalidade de obter sequências também de outras regiões do vírus, foi possível a submissão de 6 destas amostras à plataforma Ion Torrent S5™ System. Duas delas resultaram em sequências das regiões N e 3’UTR, que não somente confirmaram o agrupamento destas amostras com o gênero dos Gammacoronavirus, mas também demonstraram a proximidade destes ao vírus causador da Bronquite Infecciosa em galinhas. Este trabalho demonstrou presença de Coronavírus em amostras de aves silvestres e domésticas em diversos pontos das rotas migratórias brasileiras e a caracterização de algumas destas amostras, colaborando assim para o entendimento da epidemiologia deste agente. \"Coronavírus em aves silvestres e domésticas provenientes de diferentes regiões do Brasil\" * Deforestation and man-made changes in the environment lead to drastic changes in bird capture patterns. Continuous habitat loss and fragmentation along the corridors of major migratory routes creates bottlenecks, resulting in overpopulation and clusters of species contributing to the acquisition and creation of different pathogens in recent periods, including CoVs. Largest known RNA viruses, CoVs have a great ability to adapt new hosts and ecological niches, being the SARS and MERS viruses, both that emerged from bat viruses, the most notable examples. Despite the relevance and complexity of these agents, studies on the detection and genetic characterization of these viruses are scarce, especially in wild birds. Thus, this paper aims to detect, analyze and discuss the diversity of these agents in birds at migratory bird stop and wintering sites. Despite the relevance and the complexity of these agents, studies on the detection and genetic characterization of these viruses are scarce, especially in wild birds. Thus, this work aims to detect, analyze and discuss the diversity of these agents in birds at migratory stop and wintering sites. Therefore, 852 avian samples from different regions of Brazil were subjected to total RNA extraction followed by Reverse Transcription (RT), Polymerase Chain Reaction (PCR) and Nested-PCR reactions, with primers targeting one region. ORF 1ab coding for the viral RpRd gene, and the results were visualized by agarose gel electrophoresis. Of this total, 15 showed positive results for CoVs and in 5 of them it was possible to confirm by sequencing by the Sanger method. Additionally, 6 other CoV samples from a previous study * were analyzed to continue viral characterization studies. In the phylogenetic analysis of the ORF 1ab region sequences of these 11 samples, 3 of them were grouped with the Deltacoronavirus genus. This group is of particular importance because it has a recognized potential for transmission and emergence from poultry to mammals, such as PorCoV HKU15, associated with fatal swine outbreaks. The remaining 8 samples were grouped with the genus Gammacoronavirus, to which belongs the virus that causes infectious bronchitis disease in chickens, the best known avian CoV, which causes great economic losses, which develops in the respiratory, reproductive, gastrointestinal and kidney tract. . In order to obtain sequences also from other regions of the virus, it was possible to submit 6 of these samples to the Ion Torrent S5 ™System platform. Two of them resulted in sequences from the N and 3’UTR regions, which not only confirmed the grouping of these samples with the Gammacoronavirus genus, but also demonstrated their proximity to the infectious bronchitis virus in chickens. This work demonstrated the presence of coronaviruses in wild and domestic bird samples in several points of the Brazilian migratory routes and the characterization of some of these samples, thus contributing to the understanding of the epidemiology of this agent. \"Coronavirus in wild and domestic birds from different regions of Brazil\" *

O Torque teno vírus suíno é classificado como pertencente à família Anelloviridae gênero Iotatorquevirus que compreende as espécies: Torque teno sus virus 1a (TTSuV1a) e Torque teno sus virus 1b (TTSuV1b); e o gênero Kapatorquevirus que compreende apenas a espécie Torque teno sus virus k2 (TTSuVk2). O TTSuV foi identificado como potencial agente causador de falhas reprodutivas em porcas, como agente desencadeante nos quadros de doenças associadas ao circovírus suíno tipo 2 (PCVAD) e em associação com outros agentes como o vírus da síndrome respiratória e reprodutiva dos suínos (PRRSV), vírus da influenza suína (SIV) e Mycoplasma hyopneumoniae como co-agentes na manifestação clínica do complexo respiratório suíno (PRDC). No presente estudo foi utilizada uma PCR direcionada a região não traduzida do genoma viral (UTR), e foi investigado um total de 391 amostras divididas entre fetos abortados e mumificados, leitões natimortos, soro, fezes e pulmão de suínos de dez propriedades localizadas no Estado de São Paulo. Diferenças entre as frequências do TTSuV1 e do TTSuVk2 foram comparadas entre amostras de porcas com problemas reprodutivos (fetos abortados e mumificados e leitões natimortos), diferentes fases da criação de suínos (porcas, leitões da creche e crescimento), leitões apresentando ou não diarréia, entre animais da terminação vacinados ou não contra o PCV2 e ainda entre amostras positivas e negativas para o PCV2. Amostras positivas de pulmão de suíno e um pool de soro de Porco Monteiro do Pantanal foram submetidos a uma PCR para amplificação da ORF1 de ambos os gêneros do TTSuV, seguida pela clonagem e posterior sequenciamento para reconstrução da filogenia. Foi investigada a ocorrência de eventos de recombinação entre as amostras, pressão de seleção e propriedades da proteína relacionadas à ligação com anticorpos. O TTSuVk2 foi mais presente infectando tanto fetos abortados quanto leitões natimortos. Porcas e leitões da creche foram mais frequentemente infectados pelo TTSuV1 enquanto que leitões do crescimento pelo TTSuVk2. A coinfecção (TTSuV1 + TTSuVk2 + PCV2) foi mais frequente em amostras fecais diarreicas que nas não diarreicas. Animais da terminação apresentaram alta frequência de coinfecção (TTSuV1 + TTSuVk2), sendo ainda maior na associação com o PCV2. A filogenia construída pelo método neighborjoing baseada na sequência da ORF1 revelou existir quatro tipos do TTSuV1 (1a, 1b, 1c e 1d) e sete subtipos do TTSuVk2 (2a, 2b, 2c, 2d, 2e, 2f, 2g) circulantes nas populações de suínos domésticos e ferais do Brasil. Entre as estirpes virais circulantes foram detectados eventos de recombinação intra-hospedeiro, inter-hospedeiro, intra-genótipo e inter-genótipo. A região carboxi-terminal da proteína demonstrou agrupar características relacionadas à ligação com anticorpos. Este estudo é o primeiro a caracterizar geneticamente amostras de TTSuV circulantes em suínos domésticos e ferais do Brasil e contribui para um melhor entendimento sobre a epidemiologia do vírus. The porcine torque teno virus is classified within the Anelloviridae family, genus Iotatorquevirus comprising the species: Torque teno sus virus 1a (TTSuV1a) and Torque teno sus virus 1b (TTSuV1b). The genus Kapatorquevirus comprises only one species, the Torque teno sus virus k2 (TTSuVk2). TTSuV was identified as a potential agent of reproductive failure causes in sows, as a trigger agent associated with porcine circovirus associated diseases (PCVAD) and in combination with other agents such as porcine reproductive and respiratory syndrome (PRRSV), swine influenza virus (SIV) and Mycoplasm hyopneumoniae as a co-agent in the clinical manifestation of porcine respiratory disease complex (PRDC). In this study, PCR targeting the untranslated region (UTR) of the virus genome was used to investigate a total of 391 samples within aborted and mummified fetuses, stillborn piglets, serum, feces and lungs of pigs from ten different properties located at São Paulo State. Differences between frequencies of TTSuV1 and TTSuVk2 were compared among samples of sows with reproductive failure (aborted and mummified fetuses and stillborn piglets), different phases of pig breeding (sows, nursery and growing piglets), samples from piglets with diarrhea or not, finishing pigs vaccinated or not against PCV2 and between positive and negative samples for PCV2. Positive lung samples from pigs and a pool of sera from feral pigs were used to PCR amplification for the ORF1 of both genera of TTSuV, followed by cloning and subsequent sequencing to reconstruct the phylogeny. The occurrence of recombination events among the samples, selection pressure and protein-related antibodies binding properties was investigated. The TTSuVk2 was more frequent infecting both aborted fetuses as stillborn piglets. Sows and nursery piglets were frequently more infected by TTSuV1 while growing piglets by TTSuVk2. The coinfection (TTSuV1 + TTSuVk2 + PCV2) was more frequent in diarrheic stool samples than in the non-diarrheic. Finishing pigs showed high frequency of coinfection (TTSuV1 + TTSuVk2) and and increase of the coinfection in association with PCV2. Phylogeny constructed by neighborjoing method based on the ORF1 sequence revealed the existence of four types TTSuV1 (1a, 1b, 1c and 1d) and seven subtypes of TTSuVk2 (2a, 2b, 2c, 2d, 2e, 2f, 2g) circulating within populations of domestic and feral pigs in Brazil. Among circulating viral strains, intra-host, inter-host, intra-and inter-genotype recombination events were detected. The carboxy-terminal region of the protein showed the have characteristics related with antibodies binding activity. This study is the first to genetically characterize samples of TTSuV circulating in domestic and feral pigs from Brazil and contributes to a better understanding of the epidemiology of the virus.