Genetic characterization of the parvovirus full-length VP2 gene in domestic cats in Brazil.

Feline parvovirus (FPV) and canine parvovirus (CPV) are over 98% identical in their DNA sequences, and the new variants of CPV (2a/2b/2c) have gained the ability to infect and replicate in cats. The aim of this study was to determine the genetic diversity in the VP2 gene of parvovirus strains circulating in domestic cats in Brazil during a 10-year period (2008-2017). For parvovirus screening, specific PCR was performed, and 25 (34.7%) of 72 cats tested positive. The PCR-positive samples were further subjected to full-length VP2 sequencing (1755 bp), and eight sequences (36%) were characterized as FPV, seven (28%) as CPV-2a and (32%) nine (36%) as CPV-2b. One sequence (RJ1085/11) showing typical CPV amino acid (aa) at residues 80 R, 93 N, 103 A, 232 I, and 323 N could not be characterized at this time. The sequences in this study displayed aa changes previously described for FPV (A14T, A91S, I101T, N564S, and A568G) from cats and CPV-2a/2b (S297N and Y324L) from dogs. However, the Y324L mutation has not yet been reported in any CPV-2a/2b strains from cats. Phylogenetic analysis supported the division of these sequences into two well-defined clades, clade 1 for FPV and clade 2 for CPV2a/2b. Unusually, the sequence RJ1085/11 was grouped separately. Two recombination breakpoints were detected by Bootscan and 3Seq methods implemented in the RDP4. This study is the first report of CPV-2a/2b in cats in Brazil. The detection of FPV strains with mutations characteristic of CPV indicates that Brazilian FPV strains have undergone genetic changes.

Association between low back pain and common mental disorders in adults: systematic review / Associação entre dor lombar e transtornos mentais comuns em adultos: revisão sistemática

ABSTRACT BACKGROUND AND OBJECTIVES: Low back pain (LBP) is one of the leading causes of disability worldwide. Its etiology is multifactorial and results in losses for individuals and society. The biopsychosocial model of LBP offers a comprehensive approach to assessment and interventions. Researchers investigate the relationship between LBP and Common Mental Disorders (CMD) within this concept. This systematic review aimed to identify and analyze recent studies on the topic. CONTENTS: The study was submitted to the International Prospective Register of Systematic Reviews (registration number CRD 42020210375). The 27 articles in the corpus were selected from the Medlin/PubMed and BVS (Biblioteca Virtual em Saúde) databases from 2015 to 2022. Only those that investigated a possible relationship between CMD (anxiety or depression) and LBP in adult populations were included. Most articles in the corpus confirmed the relationship between LBP and CMD, however, gaps were identified regarding the direction of causality between the two outcomes and why this relationship was established. Methodological limitations in relation to sample selection and LBP assessment instruments used were also identified. CONCLUSION: This review highlights the need for studies on the topic with greater methodological criteria and with a design that allows causality to be assessed. Thus, it will be possible to guide clinical practice based on scientific evidence. Longitudinal studies with an emphasis on the biopsychosocial model can contribute to support specific therapeutic approaches.

RESUMO JUSTIFICATIVA E OBJETIVOS: A dor lombar (DL) é uma das principais causas mundiais de incapacidade. Trata-se de um desfecho cuja etiologia é reconhecidamente multifatorial, com prejuízos para os indivíduos e a sociedade. O modelo biopsicos-social da DL apresenta uma abordagem ampla, tanto na avaliação quanto nas intervenções. A relação entre DL e Transtorno Mental Comum (TMC) é objeto de estudo dentro da concepção desse modelo. O presente estudo teve como objetivo identificar e analisar os estudos recentes sobre esse tema. CONTEÚDO: O estudo foi submetido ao International Prospective Register of Systematic Reviews (número de registro CRD 42020210375). Os 27 artigos do corpus foram selecionados das bases de dados Medline/Pubmed e BVS (Biblioteca Virtual em Saúde) no período de 2015 a 2022. Foram incluídos apenas aqueles que investigaram possíveis relações entre TMC (ansiedade ou depressão) e DL em populações adultas. A maioria dos artigos do corpus confirmou a relação entre DL e TMC, no entanto foram identificadas lacunas sobre a direção de causalidade entre os dois desfechos e sobre os motivos pelos quais essa relação se estabeleceu. Também foram identificadas limitações metodológicas em relação à seleção da amostra e aos instrumentos de avaliação da dor lombar utilizados. CONCLUSÃO: Esta revisão destacou a necessidade de estudos sobre o tópico com maiores critérios metodológicos e com um desenho que permita avaliar a causalidade. Assim, será possível orientar a prática clínica baseando-se em evidências científicas. Estudos longitudinais com ênfase no modelo biopsicossocial podem contribuir para embasar abordagens terapêuticas específicas.

Molecular detection of porcine circovirus (PCV2 and PCV3), torque teno swine virus 1 and 2 (TTSuV1 and TTSuVk2), and histopathological findings in swine organs submitted to regular slaughter in Southeast, Brazil.

Porcine circovirus 2 and 3 (PCV2 and PCV3) and torque teno sus virus 1 and 2 (TTSuV1 and TTSuVk2) are important pathogens in pig associated with post-weaning mortality, different clinical syndromes in adults (PCVAD), and a decrease of average daily weight gain (PCV2-SI) but little is known about the infection on asymptomatic pigs. The aim of this study was to evaluate the presence of PCV2, PCV3, TTSuV1, and TTSuVk2 in swine organ samples from asymptomatic pigs slaughtered in Espírito Santo State, South-eastern Brazil, through molecular detection and histopathological analysis. Nested PCR showed the presence of PCV2 DNA in 10% (14/140), PCV3 in 13.6% (19/140), TTSuV1 in 12.9% (18/140), and TTSuVk2 in 30% (42/140) of the tissue samples. All four viruses were detected in the lung, kidney, lymph node, and liver. TTSuVk2 was detecded in 30% (42/140), PCV3 in 13.6% (19/140), TTSuV1 in 12.9% (18/140), and PCV2 in 10% (14/140) of the samples. Single infections were observed in 30.7% (43/140), while co-detections in the same tissue occurred in 15.7% (22/140). The most frequent combinations were TTSuV1/TTSuVk2 in 31.8% (7/22), PCV2/TTSuVk2 in 18.1% (4/22), and PCV2/PCV3/TTSuVk2 in 13.6% (3/22). Lymphocyte depletion was associated with TTSuVk2 infection (p = 0.0041) suggesting that TTSuVK2 plays an induction of PMWS-like lymphoid lesions in pigs. The data obtained in this study show that PCV2, PCV3, TTSuV1, and TTSuVk2 are related to infection in asymptomatic animals with different tissue lesions, and the molecular diagnosis for these pathogens should be considered in the sanitary monitoring of herds.

O circovírus suíno 2 e 3 (PCV2 e PCV3) e os Torque Teno vírus suínos 1 e 2 (TTSuV1 e TTSuVk2) são patógenos importantes na suinocultura associados a diferentes síndromes clínicas e morte de leitões pós desmame (PCVAD) e redução no ganho diário de peso (PCV2-SI). Entretanto, pouco se sabe sobre a circulação desses agentes e o impacto da infecção em porcos assintomáticos. O objetivo deste estudo foi avaliar a presença de PCV2, PCV3, TTSuV1 e TTSuVk2 em amostras de órgãos de suínos assintomáticos abatidos no estado do Espírito Santo, região sudeste do Brasil, por meio de detecção molecular e análise histopatológica. A análise tecidual por nested PCR mostrou a presença de DNA de PCV2 em 14 (10%), PCV3 em 19 (13,6%), TTSuV1 em 18 (12,9%) e de TTSuVk2 em 42 (30%) das amostras. Todos os quatro vírus foram detectados no pulmão, rim, nódulo linfático e fígado TTSuVk2 foi detectado em 30% das amostras teciduais (42/140), PCV3 em 13.6% (19/140), TTSuV1em 12.9% (18/140), e o PCV2 em 10% (14/140. Mono infecções foram observadas em 30.7% (43/140) das amostras enquanto infecções múltiplas observadas em 15.7% (22/140 das amostras de tecido). As combinações mais frequentes foram TTSuV1/TTSuVk2 em 31.8% (7/22), PCV2/TTSuVk2 em 18.1% (4/22), e PCV2/PCV3/TTSuVk2 em 13.6% (3/22). A depleção de linfócitos foi associada à infecção por TTSuVk2 (p = 0,0041) e esses achados sugerem que TTSuKV2 desempenha uma indução de lesões linfoides semelhantes a PMWS em porcos. Os dados obtidos neste estudo mostram que PCV2, PCV3, TTSuV1 e TTSuVk2 estão relacionados à infecção em animais assintomáticos com lesões teciduais diversas, e sugerem que o diagnóstico molecular para esses patógenos deve ser considerado no monitoramento sanitário dos rebanhos.

Nuclear and mitochondrial genome instability induced by fractions of ethanolic extract from Hovenia dulcis Thunberg in Saccharomyces cerevisiae strains.

Hovenia dulcis is a plant commonly used as a pharmaceutical supplement, having displayed important pharmacological properties such antigiardic, antineoplastic and hepatoprotective. The purpose of this work was investigate the cytotoxic, genotoxic and mutagenic potential from fractions of Hovenia dulcis ethanolic extract on Saccharomyces cerevisiae strains FF18733 (wild type) and CD138 (ogg1). Ethanolic extract from Hovenia dulcis leaves was fractioned using organic solvents according to increasing polarity: Hexane (1:1), dichlorometane (1:1), ethyl acetate (1:1) and butanol (1:1). Three experimental assays were performed, such as (i) inactivation of cultures; (ii) mutagenesis (canavanine resistance system) and (iii) loss of mitochondrial function (petites colonies). The findings shown a decrease in cell viability in FF18733 and CD138 strains; all fractions of the extract were mutagenic in CD138 strain; only ethyl acetate and butanol fractions increased the rate of petites colonies for CD138 strains. Ethyl acetate and n-butanol fractions induces mutagenicity, at the evaluated concentrations, in mitochondrial and genomic DNA in CD138 strain, mediated by oxidative lesions. In conclusion, it is possible to infer that the lesions caused by the extract fractions could be mediated by reactive oxygen species and might reach multiple molecular targets to cause cellular damage.

Type III atresia coli with underdeveloped colon and hydroperitoneum in a newborn foal / Atresia coli tipo III com subdesenvolvimento de cólon e hidroperitôneo em neonato equino

ABSTRACT: A 48-hour-old mixed breed pony colt was referred to the hospital because of abdominal discomfort, weak suckling reflex and prostration. During clinical investigation, supportive and symptomatic treatments were necessary, and an abdominal radiography was performed revealing a large intestine filled with feces and large amounts of gas, in addition to free fluid in the cavity. After 3 days of treatment, the foal had not yet defecated, thus exploratory abdominal surgery was indicated, but the owners declined. Therefore, the foal was humanly euthanized. Post-mortem examination revealed complete absence of the pelvic flexure. Subsequent portions of the dorsal, transverse and small colons were intensely reduced. In addition, at the abdominal cavity there was 850 ml of yellowish translucent liquid. These findings are compatible with hydroperitoneum and type III atresia coli at the pelvic flexure, associated with underdevelopment of the dorsal, transverse and small colons.

RESUMO: Um pônei mestiço de 48 horas de vida foi encaminhado ao hospital para avaliação por desconforto abdominal, diminuição do reflexo de sucção e prostração. Durante a investigação clínica, tratamentos de suporte e sintomáticos foram necessários, e radiografia abdominal foi realizada revelando cólon maior repleto de fezes e grandes quantidades de gás, além de fluido livre na cavidade. Após três dias de tratamento, o potro ainda não havia defecado, sendo a laparotomia exploratória indicada, mas os proprietários recusaram. Portanto, o potro foi humanamente eutanasiado. O exame post-mortem revelou ausência completa da flexura pélvica. Porções subseqüentes dos cólons dorsal, cólon transverso e cólon menor se apresentaram intensamente reduzidas. Além disso, na cavidade abdominal havia 850 ml de líquido translúcido amarelado. Esses achados são compatíveis com o hidroperitônio e a atresia coli tipo III na flexura pélvica, associados ao subdesenvolvimento dos cólons dorsal, transverso e menor.

Prevalencia de anaplasma spp. en humanos: revisión sistemática de la literatura entre 2000 y 2017

Introducción: La anaplasmosis es una enfermedad febril aguda transmitida por garrapatas. En humanos la especie más importante es Anaplasma phagocytophilum. Objetivo: Metanalizar la prevalencia de anaplasma en humanos reportada en la literatura científica mundial. Métodos: Revisión sistemática de la literatura según las fases Prisma con 14 estrategias de búsqueda en tres bases de datos multidisciplinarias. Se garantizó la reproducibilidad y la evaluación de la calidad metodológica. Los análisis se basaron en proporciones con sus intervalos de confianza del 95%. Resultados: Se sistematizaron 15 estudios publicados entre 2004 y 2017, la mayoría en Polonia (20%) y China (20%). El 73 % utilizó IFI y en los demás se empleó PCR anidada o Elisa. La mayoría de grupos de estudio correspondió a sujetos con exposición ocupacional o contacto con animales infectados. La prevalencia con PCR fue de 15,6% (IC de 95% = 13,1 - 18,0), con IFI de 9,3 % (IC de 95 % = 8,5 - 10,0) y con Elisa de 2,8% (IC de 95 % = 2,5 - 3,1). Conclusión: Se reportó una elevada frecuencia de infección y exposición a Anaplasma spp. en humanos, al tiempo que se identificaron puntos importantes para orientar estudios posteriores relacionados con las infecciones cruzadas, las coinfecciones y la circulación del vector.

Introduction: Anaplasmosis is an acute febrille illness transmitted through tick. In humans, the most important species is Anaplasma phagocytophilum. Objective: To meta-analyze the reported prevalence of anaplasma in humans, according to worldwide scientific literature. Method: Systematic revisión of literature according to Prisma phases with 14 strategies in search of three multidisciplinary databases. We guaranteed reproducibility and evaluation of the methodological quality. Analysis were based in proportions with confidence intervals of 95%. Results: 15 research works published between 2004 and 2017 were systematized. Most of them occurred in Poland (20%) and China (20%). 73% used IFI and the rest used PCR. Most of study groups were related to subjects with occupational exposition or contact with the infected animals. The prevalence of PCR was de 15.6% (CI of 95% = 13.1 - 18.0), with IFI of 9.3% (CI of 95% = 8.5 - 10.0). Conclusion: Studies reported a high frequency of infection and exposition to Anaplasma spp. in humans, while they identified important aspects to orientate later studies related to cross infections, coinfections and the circulation of the vector.

Introdução: a anaplasmose é uma doença febril aguda transmitida por carrapatas. Em humanos, a espécie mais importante é Anaplasma phagocytophilum. Objetivo: meta-analisar a prevalência de anaplasma em humanos relatada na literatura científica mundial. Métodos: revisão sistemática da literatura segundo as fases Prisma com 14 estratégias de busca em três bases de dados multidisciplinares. Foram garantidas a reprodutibilidade e a avaliação da qualidade metodológica. As análises se basearam em proporções com intervalos de confiança de 95 %. Resultados: foram sistematizados 15 estudos publicados entre 2004 e 2017, a maioria na Polônia (20%) e na China (20%). 73% utilizaram IFI e, nos demais, foi empregado PCR aninhado ou Elisa. A maioria de grupos de estudo correspondeu a sujeitos com exposição ocupacional ou contato com animais infectados. A prevalência com PCR foi de 15,6% (IC de 95 % = 13,1 - 18,0), com IFI de 9,3% (IC de 95% = 8,5 - 10,0) e com Elisa de 2,8% (IC de 95% = 2,5 - 3,1). Conclusão: foi relatada elevada frequência de infecção e exposição a Anaplasma spp. em humanos, ao mesmo tempo que foram identificados pontos importantes para orientar estudos posteriores relacionados com as infecções cruzadas, com as coinfecções e com a circulação do vector.

[Overview of Occupational Surveys in Brazil (2005-2015): a systematic review of the literature]. / Panorama dos Inquéritos Ocupacionais no Brasil (2005-2015):uma revisão sistemática da literatura.

Occupational Surveys are methods of collecting data from the cross-sectional studies, which are indispensable for the elaboration of public policies oriented to workers' health. The objective of this study was to perform a systematic review on articles that featured results of Occupational Surveys conducted in Brazil from 2005 to 2015. Based on the Prisma recommendation, this review selected articles indexed in Medline, Lilacs and Scielo databases. Analysis of the data consisted of the description of the Occupational Surveys according to: large regions in which the research was carried out; techniques and instruments used; topics covered; sectors of the economy in which health-work relations were investigated; and methodological criteria. Differences in the distribution of Occupational Surveys were observed regionally and by sector. The structure of the investigations, for the most part, was concerned with identifying sickness in workers. A lack of consensus regarding the methods used was revealed, which consequently prejudiced the comparison of results and monitoring of health and working conditions. Efforts are needed to build consensus and foster research in sectors and regions that are as yet scantly explored.

Inquéritos Ocupacionais são métodos de coleta de dados dos estudos transversais indispensáveis à elaboração de políticas públicas voltadas à saúde do trabalhador. O objetivo deste estudo foi realizar uma revisão sistemática sobre artigos que retratavam resultados de Inquéritos Ocupacionais conduzidos no Brasil, no período de 2005 a 2015. Baseada na recomendação Prisma, esta revisão selecionou artigos indexados em base de dados Medline, Lilacs e SciELO. A análise dos dados consistiu na descrição dos Inquéritos Ocupacionais de acordo com: grandes regiões nas quais as pesquisas foram realizadas, técnicas e instrumentos utilizados, temas abordados, setores da economia nos quais as relações saúde-trabalho foram investigadas e critérios metodológicos. Observou-se diferenças na distribuição dos Inquéritos Ocupacionais regional e setorialmente. A estrutura das investigações, em sua maioria, se interessava em identificar adoecimento em trabalhadores. Ausência de consenso no tocante aos métodos utilizados foi observada com prejuízos para a comparação de resultados e monitoramento da saúde e condições de trabalho. Esforços são necessários para a elaboração de consensos e fomento à pesquisa em setores e regiões ainda pouco exploradas.

Panorama dos Inquéritos Ocupacionais no Brasil (2005-2015): uma revisão sistemática da literatura / Overview of Occupational Surveys in Brazil (2005-2015): a systematic review of the literature

Resumo Inquéritos Ocupacionais são métodos de coleta de dados dos estudos transversais indispensáveis à elaboração de políticas públicas voltadas à saúde do trabalhador. O objetivo deste estudo foi realizar uma revisão sistemática sobre artigos que retratavam resultados de Inquéritos Ocupacionais conduzidos no Brasil, no período de 2005 a 2015. Baseada na recomendação Prisma, esta revisão selecionou artigos indexados em base de dados Medline, Lilacs e SciELO. A análise dos dados consistiu na descrição dos Inquéritos Ocupacionais de acordo com: grandes regiões nas quais as pesquisas foram realizadas, técnicas e instrumentos utilizados, temas abordados, setores da economia nos quais as relações saúde-trabalho foram investigadas e critérios metodológicos. Observou-se diferenças na distribuição dos Inquéritos Ocupacionais regional e setorialmente. A estrutura das investigações, em sua maioria, se interessava em identificar adoecimento em trabalhadores. Ausência de consenso no tocante aos métodos utilizados foi observada com prejuízos para a comparação de resultados e monitoramento da saúde e condições de trabalho. Esforços são necessários para a elaboração de consensos e fomento à pesquisa em setores e regiões ainda pouco exploradas.

Abstract Occupational Surveys are methods of collecting data from the cross-sectional studies, which are indispensable for the elaboration of public policies oriented to workers' health. The objective of this study was to perform a systematic review on articles that featured results of Occupational Surveys conducted in Brazil from 2005 to 2015. Based on the Prisma recommendation, this review selected articles indexed in Medline, Lilacs and Scielo databases. Analysis of the data consisted of the description of the Occupational Surveys according to: large regions in which the research was carried out; techniques and instruments used; topics covered; sectors of the economy in which health-work relations were investigated; and methodological criteria. Differences in the distribution of Occupational Surveys were observed regionally and by sector. The structure of the investigations, for the most part, was concerned with identifying sickness in workers. A lack of consensus regarding the methods used was revealed, which consequently prejudiced the comparison of results and monitoring of health and working conditions. Efforts are needed to build consensus and foster research in sectors and regions that are as yet scantly explored.

Feline herpesvirus 1 viral load related to environmental factors in sheltered cats / Relação entre carga viral de herpesvirus felino 1 e fatores causadores de estresse em felinos de abrigo

ABSTRACT: Shelter environment stress factors are related to FHV-1 viral reactivation. However, comparisons between conjunctival viral load and environmental factors have not been commonly evaluated. The aim of this study was to correlate FHV-1 viral load in domestic cats with and without clinical signs of conjunctivitis to shelter design in order to use FHV-1 viral load as a parameter of "health management". Cats from four different shelters underwent an ophthalmological examination. Samples were collected by rolling a DNA/RNAse-free cytobrush over the ventral conjunctival fornix and were stored in 1.5 mL sterile microtubes in 500 μL of Eagle's minimum essential medium and kept at 4 ºC. Molecular procedures were performed up to 48 hours after collection. Different routines regarding new arrivals were directly related to FHV-1 viral load. Shelters where new arrivals occurred on daily basis had the highest viral load (2.69x108 copies/µL), while those shelters where new arrivals had not occurred in the few months prior to the beginning of the study had the lowest rate (1.63x103 copies/µL). Environmental factors directly influenced FHV-1 DNA viral load. This study highlighted the need to improve the management approach in the animal shelter environment to reduce stressful situations responsible for FHV-1 reactivation and higher viral load quantification.

RESUMO: No ambiente do abrigo encontram-se fatores que geram estresse nos animais que ali residem. Esses fatores acabam por provocar a reativação do FHV-1. No entanto, comparações entre carga viral conjuntival e fatores ambientais não foram ainda avaliadas. Objetivo deste estudo foi correlacionar a carga viral de FHV-1 em felinos domésticos com e sem sinais clínicos de conjuntivite com as características dos abrigos. Assim, pode-se usar carga viral de FHV-1 como parâmetro de sanidade. Todos os gatos foram submetidos a exame clínico oftalmológico. Amostras foram coletadas com uso de escova citológica, acondicionadas em microtubos estéreis de 1,5mL contendo 500 μL de meio Eagle essencial mínimo e mantidas em 4 ºC. Análises moleculares foram realizadas no prazo de 48 horas após coleta. A rotina de entrada de novos animais estava diretamente relacionada a carga viral de FHV-1. Abrigos com entrada diária apresentaram carga viral maior (2.69x108 cópias/µL), do que abrigo onde novos animais não chegaram nos meses que antecederam a coleta (1.63x103 cópias/µL). Fatores ambientais influenciam diretamente carga viral de FHV-1. Esse estudo evidencia a necessidade de aprimorar o sistema de manejo dos abrigos de forma a reduzir situações de estresse responsáveis pela reativação de FHV-1 e consequente aumento na carga viral.

Molecular characterization of feline calicivirus variants from multicat household and public animal shelter in Rio de Janeiro, Brazil

ABSTRACT The aim of this study was to perform the molecular characterization of conserved and variable regions of feline calicivirus capsid genome in order to investigate the molecular diversity of variants in Brazilian cat population. Twenty-six conjunctival samples from cats living in five public short-term animal shelters and three multicat life-long households were analyzed. Fifteen cats had conjunctivitis, three had oral ulceration, eight had respiratory signs (cough, sneeze and nasal discharge) and nine were asymptomatic. Feline calicivirus were isolated in CRFK cells and characterized by reverse transcription PCR target to both conserved and variable regions of open reading frame 2. The amplicons obtained were sequenced. A phylogenetic analysis along with most of the prototypes available in GenBank database and an amino acid analysis were performed. Phylogenetic analysis based on both conserved and variable region revealed two clusters with an aLTR value of 1.00 and 0.98 respectively and the variants from this study belong to feline calicivirus genogroup I. No association between geographical distribution and/or clinical signs and clustering in phylogenetic tree was observed. The variants circulating in public short-term animal shelter demonstrated a high variability because of the relatively rapid turnover of carrier cats constantly introduced of multiple viruses into this location over time.

Molecular characterization of feline calicivirus variants from multicat household and public animal shelter in Rio de Janeiro, Brazil.

The aim of this study was to perform the molecular characterization of conserved and variable regions of feline calicivirus capsid genome in order to investigate the molecular diversity of variants in Brazilian cat population. Twenty-six conjunctival samples from cats living in five public short-term animal shelters and three multicat life-long households were analyzed. Fifteen cats had conjunctivitis, three had oral ulceration, eight had respiratory signs (cough, sneeze and nasal discharge) and nine were asymptomatic. Feline calicivirus were isolated in CRFK cells and characterized by reverse transcription PCR target to both conserved and variable regions of open reading frame 2. The amplicons obtained were sequenced. A phylogenetic analysis along with most of the prototypes available in GenBank database and an amino acid analysis were performed. Phylogenetic analysis based on both conserved and variable region revealed two clusters with an aLTR value of 1.00 and 0.98 respectively and the variants from this study belong to feline calicivirus genogroup I. No association between geographical distribution and/or clinical signs and clustering in phylogenetic tree was observed. The variants circulating in public short-term animal shelter demonstrated a high variability because of the relatively rapid turnover of carrier cats constantly introduced of multiple viruses into this location over time.

Molecular diversity of human parvovirus B19 during two outbreaks of erythema infectiosum in Brazil

Abstract This study was conducted to provide information on the genetic diversity of human parvovirus B19 (B19V) circulating in the municipality of Niterói, Rio de Janeiro, Southeast Brazil during 1996-2006, a period with two distinct outbreaks of B19V infection: 1999-2000 and 2004-2005. A total of 27 sera from patients with erythema infectiosum and five sera from HIV-infected patients that tested positive for B19V DNA during the study period were analyzed. To genotype B19V strains, a semi-nested PCR for partial amplification of the capsid gene was performed and sequence analysis revealed that 31 sequences belonged to subgenotype 1a (G1a) of the main genotype 1 and one sequence was characterized as subgenotype 3b (G3b). The phylogenetic tree supported the division of the G1a into two well-defined clades with 1.3% of divergence. The low diversity of the G1a strains may be explained by the fact that all patients had acute B19V infection and 30/32 sera were collected during two distinct outbreaks. The G3b strain was from an HIV-infected patient who seroconverted to anti-B19 IgG antibodies in September/2005. This is the first report of G3b in the state of Rio de Janeiro.

Molecular diversity of human parvovirus B19 during two outbreaks of erythema infectiosum in Brazil.

This study was conducted to provide information on the genetic diversity of human parvovirus B19 (B19V) circulating in the municipality of Niterói, Rio de Janeiro, Southeast Brazil during 1996-2006, a period with two distinct outbreaks of B19V infection: 1999-2000 and 2004-2005. A total of 27 sera from patients with erythema infectiosum and five sera from HIV-infected patients that tested positive for B19V DNA during the study period were analyzed. To genotype B19V strains, a semi-nested PCR for partial amplification of the capsid gene was performed and sequence analysis revealed that 31 sequences belonged to subgenotype 1a (G1a) of the main genotype 1 and one sequence was characterized as subgenotype 3b (G3b). The phylogenetic tree supported the division of the G1a into two well-defined clades with 1.3% of divergence. The low diversity of the G1a strains may be explained by the fact that all patients had acute B19V infection and 30/32 sera were collected during two distinct outbreaks. The G3b strain was from an HIV-infected patient who seroconverted to anti-B19 IgG antibodies in September/2005. This is the first report of G3b in the state of Rio de Janeiro.

Correlation between clinical signs of feline conjunctivitis and molecular detection of felid herpesvirus-1, feline calicivirus, chlamydophila felis and mycoplasma felis in cats from shelters in Rio de Janeiro / Correlação entre sinais clínicos da conjuntivite felina e a detecção molecular de felid herpesvirus-1, feline calicivirus, chlamydophila felis e mycoplasma felis em gatos de abrigos no Rio de Janeiro

Objectives: To perform molecular diagnosis of microbial agents (FHV-1, FCV, Mycoplasma felis, and Chlamydophila felis) in kittens with conjunctivitis and correlate the clinical signs with clinical severity. Material and Methods: A total of 108 conjunctival swab were collected from kittens without (G1; n = 40) and with (G2; n = 68) clinical signs of conjunctivitis. Animals from G2 group were scored from 1 (mild) to 4 (severe) according to the severity of conjunctivitis. All samples were submitted to PCR and RT-PCR. Results: FHV-1 was detected in 62/108 (57.4%) of samples, FCV in 40/108 (37.0%), M. felis in 11/108 (10.2%) and C. felis in 26/108 (24.1%). Mixed infections were detected in 39/108 (36.1%). In G1, 28/40 (70.0%) were positive for one or more agents, in G2, 58/68 (85.3%) were positive (P = 0.03). In 1, single infections by FHV-1were found in 21/40 (52.5%) samples, FCV in 2/40 (5.0%), C. felis in 1/40 (2.5%), and no pathogens were detected in 12/40 (30%) of samples, while mixed infections accounted for 29/40 (72.5%) of the cases. In G2, single FHV-1 infections were found in 31/68 (45.6%) samples, FCV in 10/68 (14.7 %), M. felis in 2/68 (3.0%) and C. felis also in 2/68 (3.0%), and no pathogens were detected in 10/68 (14.7%) samples, while mixed infections accounted for 36/68 (52.0%) of the cases. They were categorized as grade 1, 20/68 (29.4%), grade 2, 14/68 (20.6%), grade 3, 21/68 (30.9%) and grade 4, 13/68 (19.1%). The presence of FHV-1 and FCV is equally distributed among the four categories. More severe clinical signs, scores 3 and 4, are related to coinfections by C. felis and M. felis. Conclusions: FHV-1, FCV, C. felis and M. felis were identified in feline conjunctivitis. Co-infections are related to more severe cases of conjunctivitis.Molecular diagnosis is helpful to detect asymptomatic carriers and is a rapid and accurate method to determine the pathogen of feline conjunctivitis.(AU)

O objetivo deste estudo foi realizar diagnóstico molecular de agentes microbiológicos (FHV-1, FCV, Mycoplasma felis e Chlamydophila felis) em gatos filhotes e associar a presença dos patógenos à gravidade dos sinais clínicos de conjuntivite. Foram coletadas um total de 108 amostras de suabe conjuntival de filhotes felinos assintomáticos (G1; n = 40) e sintomáticos (G2; n = 68). Animais do G2 foram categorizados de 1 (leve) até 4 (grave), de acordo com o quadro clínico de conjuntivite. As 108 amostras foram submetidas à PCR e RT-PCR. O FHV-1 foi detectado em 57,4% das amostras, o FCV em 37%, o M. felis em 10,2% e o C. felis em 24,1%. Coinfecções, por sua vez, foram detectadas em 36,1%. No G1, 70% das amostras foram positivas para um ou mais patógenos. No G2, 85,3% apresentavam infecções (P = 0,03). No G1, monoinfecções por FHV-1 foram diagnosticadas em 52,5% das amostras, por FCV em 5%, por C. felis em 2,5%, e em 30% das amostras analisadas nenhum dos patógenos estudados foi encontrado. Coinfecções, por sua vez, estavam presentes em 72,5% das amostras. No G2, monoinfecções por FHV-1 foram encontradas em 45,6% das amostras, por FCV em 14,7 %, por M. felis em 3% e por C. felis também em 3%. Nenhum dos patógenos estudados foi encontrado em 14,7% das amostras analisadas. Coinfecções, responsáveis por 52% dos casos, foram categorizados como Grau 1 (29,4%), Grau 2 (20,6%), Grau 3 (30,9%) e Grau 4 (19,1%). A presença de FHV-1 e FCV está igualmente distribuída entre as quatro categorias. Os sinais clínicos mais graves (graus 3 e 4) estão relacionados a coinfecções por C. felis e M. felis. Os agentes microbiológicos FHV-1, FCV, C. felis e M. felis foram encontrados em animais com conjuntivite. Coinfecções estão relacionadas aos casos mais graves. Por fim, concluiu-se que o diagnóstico molecular, além de detectar portadores assintomáticos, é um método rápido e acurado para o diagnóstico do patógeno causador da conjuntivite felina.(AU)

Clinical aspects and weight gain reduction in swine infected with porcine circovirus type 2 and torque teno sus virus in Brazil.

Simultaneous Porcine circovirus type 2 (PCV-2) and Torque teno sus virus (TTSuV) infections have been reported around the world, generally linked to severe infections. In the present study, 257 swine plasma samples from 31 swine herds located in Brazil, were PCR screened for PCV-2 and TTSuV-1/2 and correlated with clinical data. PCV-2 was detected in 25%, followed by 38.1% and 42.4% of TTSuV-1 and TTSuV-2, respectively. Co-infections of two or three viruses were found in 32.3% of samples. PCV-2 was more frequently detected in the growing (p=0.030) and finishing phases (p=0.0005) while TTSuV-2 in the nursery (p=0.009). Only TTSuV-1 was statistically associated to clinical disease (multiple signs), in combination or not with PCV-2 or TTSuV-2 (p=0.015). PCV-2/TTSuV co-infections were more frequently related to weight gain reduction in comparison to mono-infections (p=0.049) and no-infections (p=0.027), and also in animals with (p=0.011) or without (p=0.037) clinical signs, being the nursery the most affected phase (p=0.025). Our results uphold the pathogenic potential of TTSuV in naturally infected pigs and the clinical/economical impact of this agent, especially in co-infections. Studies addressing the physiopathological mechanisms of simultaneous infections are needed.

Cynomolgus monkeys (Macaca fascicularis) experimentally infected with B19V and hepatitis A virus: no evidence of the co-infection as a cause of acute liver failure.

This study was conducted to analyse the course and the outcome of the liver disease in the co-infected animals in order to evaluate a possible synergic effect of human parvovirus B19 (B19V) and hepatitis A virus (HAV) co-infection. Nine adult cynomolgus monkeys were inoculated with serum obtained from a fatal case of B19V infection and/or a faecal suspension of acute HAV. The presence of specific antibodies to HAV and B19V, liver enzyme levels, viraemia, haematological changes, and necroinflammatory liver lesions were used for monitoring the infections. Seroconversion was confirmed in all infected groups. A similar pattern of B19V infection to human disease was observed, which was characterised by high and persistent viraemia in association with reticulocytopenia and mild to moderate anaemia during the period of investigation (59 days). Additionally, the intranuclear inclusion bodies were observed in pro-erythroblast cell from an infected cynomolgus and B19V Ag in hepatocytes. The erythroid hypoplasia and decrease in lymphocyte counts were more evident in the co-infected group. The present results demonstrated, for the first time, the susceptibility of cynomolgus to B19V infection, but it did not show a worsening of liver histopathology in the co-infected group.

Cynomolgus monkeys (Macaca fascicularis) experimentally infected with B19V and hepatitis A virus: no evidence of the co-infection as a cause of acute liver failure

This study was conducted to analyse the course and the outcome of the liver disease in the co-infected animals in order to evaluate a possible synergic effect of human parvovirus B19 (B19V) and hepatitis A virus (HAV) co-infection. Nine adult cynomolgus monkeys were inoculated with serum obtained from a fatal case of B19V infection and/or a faecal suspension of acute HAV. The presence of specific antibodies to HAV and B19V, liver enzyme levels, viraemia, haematological changes, and necroinflammatory liver lesions were used for monitoring the infections. Seroconversion was confirmed in all infected groups. A similar pattern of B19V infection to human disease was observed, which was characterised by high and persistent viraemia in association with reticulocytopenia and mild to moderate anaemia during the period of investigation (59 days). Additionally, the intranuclear inclusion bodies were observed in pro-erythroblast cell from an infected cynomolgus and B19V Ag in hepatocytes. The erythroid hypoplasia and decrease in lymphocyte counts were more evident in the co-infected group. The present results demonstrated, for the first time, the susceptibility of cynomolgus to B19V infection, but it did not show a worsening of liver histopathology in the co-infected group.

Detection and molecular characterization of caliciviruses (vesivirus and norovirus) in an outbreak of acute diarrhea in kittens from Brazil.

Feline caliciviruses (FCVs) have occasionally been described in cats in association with enteric disease, but an etiological role for these viruses in acute gastroenteritis is still unclear. In this study, molecular characterization of FCV and feline norovirus (FNoV) was undertaken using real-time PCR (RT-PCR) and sequence analysis of the ORF1 region in fecal specimens from 29 diarrheic cats. The specimens were also screened for parvovirus, coronavirus, astrovirus and group A rotavirus. A quantitative one step RT-PCR was also performed to detect and quantitate NoV genogroup IV and the role of these animal caliciviruses in feline gastroenteritis was investigated. This is the first description of enteric FCV and FNoV in South America.

Genetic diversity of Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil.

The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.

Genetic diversity of Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil / Diversidade genética de cepas de Ehrlichia canis encontradas em cães naturalmente infectados no Rio de Janeiro, Brasil

The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of 1the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.

O objetivo deste estudo foi caracterizar as cepas de Ehrlichia canis em cães naturalmente infectados no Rio de Janeiro, Brasil. Além disso, os achados clínicos e hematológicos observados nos cães foram relatados. O gene 16S rRNA foi utilizado como alvo da PCR para fins diagnósticos, e os genes TRP19 e TRP36 para avaliar a diversidade genética. Quinze amostras foram positivas para E. canis. PCR para o gene TRP19 produziu 11 amplicons (11/15) que foram clonados no pGEM-T easy vector para sequenciamento. A comparação das sequências completas do gene TRP19 com outras sequências depositadas no GenBank revelou uma alta identidade. Duas amostras (56C e 70C) após o ensaio da PCR, tendo como alvo o gene TRP36, geraram sequências, e a análise filogenética mostrou que a cepa 56C foi agrupada com a cepa Cuiabá 16, que é uma cepa híbrida, formada pelo genogrupo Brasileiro e o genogrupo US; e a cepa 70C agrupou com as outras cepas do genogrupo US, sugerindo a existência de pelo menos dois genogrupos de E. canis no Rio de Janeiro (US e Brasileiro). Esses animais apresentaram manifestações clínicas e hematológicas distintas, e diferentes genótipos podem expressar novos fenótipos, resultando em diferentes formas de apresentação da doença e fazendo com que o diagnóstico seja mais complexo.