Comparative evaluation of HPV genotyping: A study on the performance concordance between Anyplex II HPV28 detection and Linear Array genotyping tests in nationwide studies in Brazil.

BACKGROUND: Advances in laboratory techniques for HPV diagnosis necessitate a thorough assessment of the efficiency, replicability, sensitivity, and specificity of those methods. This study aims to validate and compare HPV detection/genotyping using the Anyplex™ II HPV28 Detection assay (Seegene) assay and the Linear Array HPV Genotyping test (Roche Diagnostics) on genital samples for use in epidemiological studies. METHODS: From 6,388 penile and cervical DNA samples collected in the POP-Brazil, 1,745 were randomly selected to be included in this study. The samples were submitted to HPV detection and genotyping following the manufacturers' protocols. DNA was genotyped using the Anyplex™ II HPV28 Detection kit (Seegene), and the results were compared to those obtained using the Linear Array HPV Genotyping test (Roche Diagnostics). Concordance of HPV genotyping results was assessed by the percentage agreement and Cohen's kappa score (κ). RESULTS: The agreement between the two methodologies was deemed good for HPV detection (κ = 0.78). Notably, Anyplex™ II HPV28 demonstrated enhanced capability in detecting a broader spectrum of genotypes compared to Linear Array. CONCLUSION: Anyplex™ II HPV28 exhibited comparable results to the Linear Array assay in clinical specimens, showcasing its potential suitability for a diverse array of research applications requiring the detection and genotyping of HPV. The study supports the utility of Anyplex™ II HPV28 as an effective tool for HPV screening in epidemiological studies, emphasizing its robust performance in comparison to established diagnostic tests.

Incipient Parallel Evolution of SARS-CoV-2 Deltacron Variant in South Brazil.

With the coexistence of multiple lineages and increased international travel, recombination and gene flow are likely to become increasingly important in the adaptive evolution of SARS-CoV-2. These processes could result in genetic introgression and the incipient parallel evolution of multiple recombinant lineages. However, identifying recombinant lineages is challenging, and the true extent of recombinant evolution in SARS-CoV-2 may be underestimated. This study describes the first SARS-CoV-2 Deltacron recombinant case identified in Brazil. We demonstrate that the recombination breakpoint is at the beginning of the Spike gene. The 5' genome portion (circa 22 kb) resembles the AY.101 (Delta), and the 3' genome portion (circa 8 kb nucleotides) is most similar to the BA.1.1 (Omicron). Furthermore, evolutionary genomic analyses indicate that the new strain emerged after a single recombination event between lineages of diverse geographical locations in December 2021 in South Brazil. This Deltacron, AYBA-RS, is one of the dozens of recombinants described in 2022. The submission of only four sequences in the GISAID database suggests that this lineage had a minor epidemiological impact. However, the recent emergence of this and other Deltacron recombinant lineages (XD, XF, and XS) suggests that gene flow and recombination may play an increasingly important role in the COVID-19 pandemic. We explain the evolutionary and population genetic theory that supports this assertion, concluding that this stresses the need for continued genomic surveillance. This monitoring is vital for countries where multiple variants are present, as well as for countries that receive significant inbound international travel.

Genomic evidence of SARS-CoV-2 reinfection cases in southern Brazil.

Cases of reinfection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been reported worldwide. We investigated reinfection cases in a set of more than 30,000 samples, and the SARS-CoV-2 genomes from selected samples from four patients with at least two positive diagnoses with an interval ≥ 45 days between tests were sequenced and analyzed. Comparative genomic and phylogenetic analysis confirmed three reinfection cases and suggested that the fourth one was caused by a virus of the same lineage. Viral sequencing is crucial for understanding the natural course of reinfections and for planning public health strategies for management of COVID-19.

Viral load is associated with mitochondrial dysfunction and altered monocyte phenotype in acute severe SARS-CoV-2 infection.

Monocytes play a major role in the initial innate immune response to SARS-CoV-2. Although viral load may correlate with several clinical outcomes in COVID-19, much less is known regarding their impact on innate immune phenotype. We evaluated the monocyte phenotype and mitochondrial function in severe COVID-19 patients (n = 22) with different viral burden (determined by the median of viral load of the patients) at hospital admission. Severe COVID-19 patients presented lower frequency of CD14 + CD16- classical monocytes and CD39 expression on CD14 + monocytes, and higher frequency of CD14 + CD16 + intermediate and CD14-CD16 + nonclassical monocytes as compared to healthy controls independently of viral load. COVID-19 patients with high viral load exhibited increased GM-CSF, PGE-2 and lower IFN-α as compared to severe COVID-19 patients with low viral load (p < 0.05). CD14 + monocytes of COVID-19 patients with high viral load presented higher expression of PD-1 but lower HLA-DR on the cell surface than severe COVID-19 patients with low viral load. All COVID-19 patients presented decreased monocyte mitochondria membrane polarization, but high SARS-CoV-2 viral load was associated with increased mitochondrial reactive oxygen species. In this sense, higher viral load induces mitochondrial reactive oxygen species generation associated with exhaustion profile in CD14 + monocytes of severe COVID-19 patients. Altogether, these data shed light on new pathological mechanisms involving SARS-CoV-2 viral load on monocyte activation and mitochondrial function, which were associated with COVID-19 severity.

Open-source real-time quantitative RT-PCR-based on a RNA standard for the assessment of SARS-CoV-2 viral load.

BACKGROUND: Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) target genes by molecular methods has been chosen as the main approach to identify individuals with Coronavirus disease 2019 (COVID-19) infection. OBJECTIVES: In this study, we developed an open-source RNA standard-based real-time quantitative RT-PCR (RT-qPCR) assay for quantitative diagnostics of SARS-CoV-2 from nasopharynx, oropharynx, saliva and plasma samples. METHODS AND FINDINGS: We evaluated three SARS-CoV-2 target genes and selected the RNA-dependent RNA polymerase (RdRp) gene, given its better performance. To improve the efficiency of the assay, a primer gradient containing 25 primers forward and reverse concentration combinations was performed. The forward and reverse primer pairs with 400 nM and 500 nM concentrations, respectively, showed the highest sensitivity. The LOD95% was ~60 copies per reaction. From the four biological matrices tested, none of them interfered with the viral load measurement. Comparison with the AllplexTM 2019-nCoV assay (Seegene) demonstrated that our test presents 90% sensitivity and 100% specificity. MAIN CONCLUSIONS: We developed an efficient molecular method able to measure absolute SARS-CoV-2 viral load with high replicability, sensitivity and specificity in different clinical samples.

Emergence of the novel SARS-CoV-2 lineage VUI-NP13L and massive spread of P.2 in South Brazil.

In this study, we analyzed 340 whole genomes of SARS-CoV-2, which were sampled between April and November 2020 in 33 cities of Rio Grande do Sul, South Brazil. We demonstrated the circulation of two novel emergent lineages, VUI-NP13L and VUI-NP13L-like, and five major lineages that had already been assigned (B.1.1.33, B.1.1.28, P.2, B.1.91, B.1.195). P.2 and VUI-NP13L demonstrated a massive spread in October 2020. Constant and consistent genomic surveillance is crucial to identify newly emerging SARS-CoV-2 lineages in Brazil and to guide decision making in the Brazilian Public Healthcare System.

Effectiveness of a universal vaccination program with an HPV quadrivalent vaccine in young Brazilian women.

We examined human papillomavirus (HPV) vaccine effectiveness in a nationwide sample of women aged 16 to 25 years who utilized the public health system in Brazil. This was a cross-sectional, multicentric survey conducted between September 2016 and November 2017 (POP-Brazil Study). A total of 5,945 young adult women were recruited from 119 public primary care units from all 27 federative units of Brazil by trained health professionals. The participants participated in a face-to-face interview and provided biological samples for genital HPV analysis. HPV genotyping was performed using a Linear Array HPV genotyping test in a central laboratory. Sampling weights were applied to the data. Overall, 11.92% (95% CI 10.65, 13.20) of the participants reported having been vaccinated. The frequency of vaccination was highest in 16- to 17-year-old women, with a decreasing vaccination rate with increasing age, and vaccinated women were more likely to belong to the high socioeconomic status group. The use of a quadrivalent vaccine decreased the HPV types 6, 11, 16, and 18 by 56.78%, from 15.64% in unvaccinated women to 6.76% in vaccinated women (P < 0.01), even after adjustment for age. Those who received the vaccine had lower HPV 16 (2.34% in vaccinated vs 8.91% in unvaccinated, P < 0.01) and 6 rates (2.06% vs 5.77%, P < 0.01). Additionally, a higher rate of high-risk HPV types other than HPV 16 and 18 (40.47% in vaccinated vs 32.63% in unvaccinated, P < 0.01) was observed. In conclusion, the results of this study support the effectiveness of HPV vaccination in Brazil. Continuous surveillance must be assured to monitor the HPV infection rate in the vaccination era.

Open-source real-time quantitative RT-PCR-based on a RNA standard for the assessment of SARS-CoV-2 viral load

BACKGROUND Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) target genes by molecular methods has been chosen as the main approach to identify individuals with Coronavirus disease 2019 (COVID-19) infection. OBJECTIVES In this study, we developed an open-source RNA standard-based real-time quantitative RT-PCR (RT-qPCR) assay for quantitative diagnostics of SARS-CoV-2 from nasopharynx, oropharynx, saliva and plasma samples. METHODS AND FINDINGS We evaluated three SARS-CoV-2 target genes and selected the RNA-dependent RNA polymerase (RdRp) gene, given its better performance. To improve the efficiency of the assay, a primer gradient containing 25 primers forward and reverse concentration combinations was performed. The forward and reverse primer pairs with 400 nM and 500 nM concentrations, respectively, showed the highest sensitivity. The LOD95% was ~60 copies per reaction. From the four biological matrices tested, none of them interfered with the viral load measurement. Comparison with the AllplexTM 2019-nCoV assay (Seegene) demonstrated that our test presents 90% sensitivity and 100% specificity. MAIN CONCLUSIONS We developed an efficient molecular method able to measure absolute SARS-CoV-2 viral load with high replicability, sensitivity and specificity in different clinical samples.

Sex differences in the prevalence and determinants of HPV-related external genital lesions in young adults: a national cross-sectional survey in Brazil.

BACKGROUND: External genital lesions (EGL) are the most common sexually transmitted infections (STIs). We aimed to evaluate the prevalence, determinants and sex differences in EGL among young adults from Brazil. METHODS: Overall, 7694 participants (aged 16 to 25 years) underwent an interview, genital examination and sampling for HPV genotyping. RESULTS: The prevalence of EGL was 4.08% (234) and is more frequent in men (5.72%) than women (2.31%) (p <  0.001). Genital lesions were significantly associated with male sex, infection by high-risk and multiple HPV types, having more than two sexual partners in the last year, smoking status and the presence of other STI. While alcohol use was associated with a higher prevalence of EGL in women, same-sex sexual relationship increase the prevalence in men. In the EGL group, 67.79% (p = 0.032) were positive for HPV infection and the types HPV6 and HPV11 were the most prevalent ones. CONCLUSION: The prevalence of EGL in young adults was consistently high, and most cases were associated with genital HPV infection and STIs. Although men have a higher prevalence, both sexes share most genital lesion determinants. The promotion of sexual education and vaccination especially focus in young men, who are usually outside the targets of primary health care programmes, can prevent EGL in Brazilian young adults.

Identification of a murine cell line that distinguishes virulent from attenuated isolates of the morbillivirus Peste des Petits Ruminants, a promising tool for virulence studies.

Comprehensive pathogenesis studies on Peste des Petits Ruminants virus (PPRV) have been delayed so far by the absence of a small animal model reproducing the disease or an in vitro biological system revealing virulence differences. In this study, a mouse 10T1/2 cell line has been identified as presenting different susceptibility to virulent and attenuated PPRV strains. As evidenced by immunofluorescence test and RT-PCR, both virulent and attenuated PPR viruses penetrated and initiated the replication cycle in 10T1/2 cells, independently of the presence of the SLAM goat receptor. However, only virulent strains successfully completed their replication cycle while the vaccine strains did not. Since 10T1/2 cells are interferon-producing cells, the role of the type I interferon (type I IFN) response on this differentiated replication between virulent and attenuated strains was verified by stimulation or repression. Modulation of the type I IFN response did not improve the replication of the vaccine strains, indicating that other cell factor(s) not yet established may hinder the replication of attenuated PPRV in 10T1/2. This 10T1/2 cell line can be proposed as a new in vitro tool for PPRV-host interaction and virulence studies.

STOP HPV study protocol: a nationwide case-control study of the association between oropharyngeal cancer and human papillomavirus (HPV) infection in Brazil.

INTRODUCTION: Human papillomavirus (HPV) is the most common sexually transmitted infection and is associated with several types of cancer. The number of cases of HPV-associated head and neck squamous cell carcinomas (HNSCCs), especially oropharyngeal carcinomas, has increased significantly in recent years despite decreased tobacco smoking rates. Currently, no data concerning the risk factors and prevalence of HPV in HNSCC patients in all regions of Brazil are available, making it difficult to promote advances in this field of public health. Therefore, our goal is to determine the impact of infection by HPV, including HPVs with different genotypes, on head and neck cancer and the risk factors associated with the development of head and neck cancer in all regions of Brazil. METHODS AND ANALYSIS: This is a case-control study that will include 622 patients and 622 controls from all regions of Brazil. A questionnaire will be applied to gather information on sociodemographic, behavioural and health factors. Oral, cervical or penile/scrotal, and anal specimens and serum samples will be collected from all participants. Formalin-fixed paraffin-embedded tissue from tumour biopsies will be analysed only in the case group. Molecular and serological analyses will be performed to evaluate the presence and role of HPV in the development of head and neck cancer. ETHICS AND DISSEMINATION: This project was approved by the research ethical committee of the proposing institution (Hospital Moinhos de Vento, number 2.852.060). Ethical approval from the collaborators is currently under evaluation and is not yet complete. The results of this study will be presented at meetings with the Brazilian Ministry of Health through technical reports and to the scientific community at national and international events, with subsequent publication of scientific articles.

Evaluating sexual health in sex workers and men who have sex with men: the SMESH cross-sectional protocol study.

INTRODUCTION: Human papillomavirus (HPV) infection is transmitted through skin-to-skin contact, and vaginal and anal sex are the most common transmission routes. Sex workers and men who have sex with men (MSM) are more exposed to the virus, and therefore, a higher frequency of this infection would be expected. The prevalence of HPV infection types and the forms and factors of transmission must be investigated to control infection-related outcomes. This protocol study will be the first nationwide study with a uniform methodology to evaluate HPV prevalence of and infection types among sex workers and MSM in Brazil. METHODS AND ANALYSIS: This multicentre cross-sectional study will be conducted with a respondent-driven sampling method to recruit 1174 sex workers and 1198 MSM from all regions of Brazil. The study will consist of preliminary interviews to verify the eligibility criteria and characterise the network size as well as a second questionnaire to obtain sociodemographic, behavioural and sexual information. Specimens from the oral cavity and anal and cervical or penile/scrotal sites will be collected. All HPV samples will be processed in a certified central laboratory. Other sexually transmitted infections will be evaluated by interview and by rapid testing for HIV and syphilis. Strict quality control will be conducted using different procedures, including the training and certification of the health professionals responsible for acquiring data and monitoring visits. ETHICS AND DISSEMINATION: The project was approved by the research ethics committee of the main institution and the corresponding ethics committees of the recruitment sites. Due to the literature gap on the sexual health of sex workers and MSM and the intense stigma surrounding these populations, a critical analysis of the study results will contribute to epidemiological knowledge and will be useful for the development of strategies against virus morbidities.

Absence of A3Z3-Related Hypermutations in the env and vif Proviral Genes in FIV Naturally Infected Cats.

Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3; A3) proteins comprise an important family of restriction factors that produce hypermutations on proviral DNA and are able to limit virus replication. Vif, an accessory protein present in almost all lentiviruses, counteracts the antiviral A3 activity. Seven haplotypes of APOBEC3Z3 (A3Z3) were described in domestic cats (hap I⁻VII), and in-vitro studies have demonstrated that these proteins reduce infectivity of vif-defective feline immunodeficiency virus (FIV). Moreover, hap V is resistant to vif-mediated degradation. However, studies on the effect of A3Z3 in FIV-infected cats have not been developed. Here, the correlation between APOBEC A3Z3 haplotypes in domestic cats and the frequency of hypermutations in the FIV vif and env genes were assessed in a retrospective cohort study with 30 blood samples collected between 2012 and 2016 from naturally FIV-infected cats in Brazil. The vif and env sequences were analyzed and displayed low or undetectable levels of hypermutations, and could not be associated with any specific A3Z3 haplotype.

Avaliação funcional de pacientes internados na Unidade de Terapia Intensiva adulto do Hospital Universitário de Canoas / Evaluación funcional de pacientes internados en la unidad de cuidados intensivos adultos de Hospital Universitario de Canoas / Functional assessment of inpatients in the Intensive Care Unit of the University Hospital of Canoas

RESUMO A sobrevida de pacientes críticos tem aumentado com o tempo. No entanto, a imobilidade e o tempo de internação estão contribuindo para o seu declínio funcional e da sua qualidade de vida. O objetivo do estudo foi avaliar a independência funcional dos pacientes internados na Unidade de Terapia Intensiva (UTI) Adulto do Hospital Universitário de Canoas. Pesquisa de coorte prospectiva executada de fevereiro a dezembro de 2016. Os pacientes foram avaliados quanto à capacidade funcional, força muscular, força de preensão palmar, mobilidade, equilíbrio e marcha. Foram avaliados 90 pacientes com média de idade de 59,6±16,1 anos, com predominância do gênero masculino (51,1%). A mediana do tempo de internação na UTI foi de 5 (3-9) dias, e de internação hospitalar de 13 (10-20) dias. Houve melhora significativa nos resultados de capacidade funcional (p<0,001), mobilidade (p=0,004) e equilíbrio (p=0,009). Os pacientes internados apresentaram um declínio funcional (com relação à normalidade) nos momentos avaliados. Entretanto, houve melhora nos valores até momento da alta hospitalar.

RESUMEN La sobrevida de pacientes críticos ha aumentado con el tiempo. Sin embargo, la inmovilidad y el tiempo de internación están contribuyendo a su declive funcional y su calidad de vida. El objetivo del estudio ha sido evaluar la independencia funcional de los pacientes internados en la Unidad de Cuidados Intensivos (UCI) Adulto de Hospital Universitário de Canoas. Investigación de cohorte prospectiva ejecutada de febrero a diciembre de 2016. Los pacientes han sido evaluados en cuanto a la capacidad funcional, fuerza muscular, fuerza de prensión de mano, movilidad, equilibrio y marcha. Se evaluaron 90 pacientes con una media de edad de 59.6±16.1 años, con predominio del género masculino (51.1%). La mediana del tiempo de internación en la UCI ha sido de 5 (3-9) días, y de internación hospitalaria de 13 (10-20) días. Se observó una mejora significativa en los resultados de capacidad funcional (p<0.001), movilidad (p=0.004) y equilibrio (p=0.009). Los pacientes internados presentaron un declive funcional (con relación a la normalidad) en los momentos evaluados. Sin embargo, hubo mejora en los valores hasta el momento del alta hospitalaria.

ABSTRACT Survival of critically ill patients has increased over time. However, immobility and length of hospital stay contribute to these patient's functional decline and reduction in quality of life. To assess the functional independence of patients admitted to the Intensive Care Unit (ICU) of the University Hospital of Canoas, a prospective cohort study was performed from February to December 2016. Functional capacity, muscle strength, hand grip strength, mobility, balance, and gait were assessed. 90 patients aged on average 59.6±16.1 years old were assessed, with a predominance of male individuals (51.1%). The median of length of stay in the ICU was 5 (3-9) days and the median of hospital stay was 13 (10-20) days. Functional capacity (p<0.001), mobility (p=0.004), and balance (p=0.009) significantly improved. Hospitalized patients showed functional decline (compared to normative values) in all assessments. However, all scores improved after hospital discharge.

Phylodynamics of the Brazilian feline immunodeficiency virus.

Feline immunodeficiency virus (FIV), like other retroviruses, displays large genomic divergence when different isolates are compared. In this study, 31 FIV positive samples of domestic cats from Porto Alegre, RS, Brazil were used aiming at a detailed genomic characterization and a better understanding of the molecular epidemiology of the virus in Brazil. The proviral env genes were partially amplified, sequenced and compared with another 237 sequences from different continents. We identified several Brazilian highly supported clades (A, B1, B2, C and D) that suggest independent events of introduction of FIV in Brazil. Forty six reference-sequences from the GenBank were used with our 31 sequences to infer the virus subtypes. Our sequences belong to the subtype B and three of them result from a recombination with the previously described subtype F. The other 28 Brazilian samples belonging to subtype B and another 46 Brazilian sequences from the GenBank were used to estimate the time to the most recent common ancestor of each Brazilian clade, using a Bayesian approach and a relaxed molecular clock model. The analyses of Brazilian sequences suggest several different entries of the virus in the Brazilian cat population between 1981 and 1991.

Molecular detection and characterization of BK and JC polyomaviruses in urine samples of renal transplant patients in Southern Brazil.

The human polyomaviruses JC (JCPyV) and BK (BKPyV) are widespread in the human population. Following the primary infection, virus reactivation may lead to nephropathy and graft rejection in renal transplant patients. This study was carried out to access the presence of BKPyV and JCPyV DNA in urine samples collected from renal transplant patients (n = 92) and healthy individuals (n = 88) in Porto Alegre, Rio Grande do Sul. The samples were submitted to a nested PCR. A significantly higher frequency (P < 0.001) of BKPyV was found in renal transplant patients (65.2%) in comparison to the control group (32.9%). JCPyV was detected equally in both groups. Phylogenetic analysis of both BKPyV and JCPyV amplicons demonstrates the presence of the BKPyV subtypes I and II, whereas for JCPyV, four different groups are found (1, 2, 3, and 4).

Detection of human adenovirus, rotavirus and enterovirus in water samples collected on dairy farms from Tenente Portela, Northwest of Rio Grande do Sul, Brazil

Viral gastroenteritis and other waterborne diseases are a major concern for health in Brazil. A number of studies were conducted about the presence of viruses on water samples from Brazilian areas. However, the knowledge about the occurrence of viral contamination of drinking water sources in rural settings of the country is insufficient. On the present work, 15 samples from 5 dairy farms located at the municipality of Tenente Portela were collected and analysed for the presence of human adenoviruses (HAdV), as well as human enteroviruses (EV) and rotaviruses (RV). HAdV was present on 66.66% of the water samples, and have been found in all samples from artesian wells and springs, which are used as sources of drinking water for the individuals inhabiting those farms. EV and RV found only in one sample each. The detection rates of HAdV on the water from these dairy farms are alarming and point towards a situation of elevated environmental contamination by fecal microorganisms of human origin and poor basic sanitation conditions.

Detecção molecular de vírus da bronquite infecciosa em plantéis de avós, matrizes e frangos de corte no Rio Grande do Sul e Mato Grosso / Molecular detection of infectious bronchitis virus in flocks of grandmothers, mothers and broiler chicken in Rio Grande do Sul and Mato Grosso

O vírus da Bronquite Infecciosa das galinhas (VBI) pertence ao grupo 3 da família Coronaviridae e é o causador de desordens respiratórias e renais em frangos de corte. A vacinação com vacinas vivas é praticada em matrizes e avós e muitas vezes também nos plantéis destinados ao abate. As vacinas utilizadas no Brasil são usualmente do sorogrupo Massachusetts e baseadas nas amostras H120 e H52. É comum que após a vacinação o vírus vacinal seja detectado por isolamento em ovos embrionados ou por métodos moleculares por até 4 semanas. Após essa data, normalmente, não há detecção de vírus e o VBI, quando encontrado, pode representar recirculação do vírus vacinal no plantel ou a introdução de uma nova cepa do vírus. No presente estudo, para avaliar a circulação do vírus em plantéis de frangos e reprodutoras nos estados do Rio Grande do Sul e Mato Grosso do Sul, foram coletadas 240 traqueias e rins de aves de 48 plantéis, sendo (20 exemplares/4 plantéis) de avós, (80 exemplares/16 plantéis) de matrizes e (140 exemplares/28 plantéis) de frangos de corte, as quais foram analisadas em misturas de cinco amostras. Todos os animais eram vacinados e as amostras foram coletadas ao redor de 2 a 48 semanas após a vacinação. A presença de VBI foi determinada com auxílio de uma reação em cadeia da polimerase tipo nested, direcionada ao gene da proteína S1, padronizada neste estudo. Das 48 amostras testadas, 14 resultaram positivas: cinco foram oriundas de aves vacinadas há menos de quatro semanas na data da coleta e nove eram de amostras de aves vacinadas há mais de quatro semanas, o que pode ser devido à recirculação do vírus vacinal ou mesmo introdução de vírus selvagem nos plantéis.

Infectious bronchitis virus (IBV, Avian Coronavirus) from chickens belongs to group 3 of the family Coronaviridae and causes respiratory and renal disorders in broilers. Vaccination using live vaccines is generally performed in mothers and grandmothers, as well as often in flocks for slaughter. The vaccines used in Brazil are usually from serogroup Massachusetts and based on standard samples of the virus at passages H120 and H52. It is common that after vaccination the vaccine virus is detected by isolation in embryonated eggs or by molecular methods for up to four weeks. After, there is usually no virus detection and any IBV found may represent recirculation of the vaccine virus in the flock or the introduction of a new strain. In this study, to evaluate the circulation of the virus in poultry flocks and breeders in the state of Rio Grande do Sul and Mato Grosso do Sul, 240 samples were collected from tracheas and kidneys of birds from 48 flocks, and (20 biological samples / 4 flocks) from grandmothers (80 samples/16 flocks) and mothers (140 samples/28 flocks) from broilers, which were analyzed in pools of five samples. All animals were vaccinated and samples were collected around 2-48 weeks after vaccination. The presence of IBV was determined with the aid of a polymerase chain reaction "nested" gene-directed protein S1, standardized in this study. From the 48 samples tested, 14 were positive: 5 were from birds vaccinated after less than 4 weeks and 9 were from birds vaccinated more than four weeks should be wild viruses or represent the recirculation of the vaccine virus.

Presence of Torque teno virus (TTV) in tap water in public schools from Southern Brazil.

Torque teno virus (TTV) was surveyed in tap water collected in schools from three municipalities located in the south of Brazil. TTV genomes were found in 11.7 % (4/34) of the samples. TTV DNA was detected in 10.5 % (2/19) of the samples collected at the city of Caxias do Sul and in 25 % (2/8) of the samples from Pelotas. Those cities have a low rate of sewage treatment. All samples from Santa Cruz do Sul, which has nearly 92 % of its sewage treated, were negative. These results suggest that the amount of sewage treated may have an effect on the detection rates of TTV DNA in drinking water in a given urban area, showing a mild negative correlation (r = -0.76), when comparing the percentage of sewage treatment to the detection of TTV genomes. The detection rate of TTV was also compared with Escherichia coli, showing a strong correlation (r = 0.97), indicating that TTV may be a suitable marker of fecal contamination.

Detection of human adenovirus, rotavirus and enterovirus in water samples collected on dairy farms from Tenente Portela, Northwest of Rio Grande do Sul, Brazil.

Viral gastroenteritis and other waterborne diseases are a major concern for health in Brazil. A number of studies were conducted about the presence of viruses on water samples from Brazilian areas. However, the knowledge about the occurrence of viral contamination of drinking water sources in rural settings of the country is insufficient. On the present work, 15 samples from 5 dairy farms located at the municipality of Tenente Portela were collected and analysed for the presence of human adenoviruses (HAdV), as well as human enteroviruses (EV) and rotaviruses (RV). HAdV was present on 66.66% of the water samples, and have been found in all samples from artesian wells and springs, which are used as sources of drinking water for the individuals inhabiting those farms. EV and RV found only in one sample each. The detection rates of HAdV on the water from these dairy farms are alarming and point towards a situation of elevated environmental contamination by fecal microorganisms of human origin and poor basic sanitation conditions.