Toxoplasma gondii infection in pig intended for human consumption: seroprevalence, risk factors and influence of biosecurity measures.

A serologic and epidemiologic study was carried out in order to determinate herd and animal seroprevalence and associated factors for Toxoplasma gondii in commercial pigs from Espírito Santo state, Brazil. Blood samples were collected from 416 pigs from 55 producer farms in 27 municipalities. An indirect immunofluorescent assay (IFA) was performed to estimate the seroprevalence of T. gondii and identify the associated risk factors using a questionnaire. The T. gondii antibody prevalence rate in commercial swine herds was 15.4% (64/416) using a cutoff of 1:64. The seropositivity for T. gondii was related to the presence of cats, water origin and age of swine in the increase of seroprevalence, and the existence of internal isolation fences and use of composting chambers as protective factors. To the best of our knowledge, this is the first study to report anti- T. gondii antibodies in the serum of pigs in the state of Espírito Santo, Brazil. This finding is important to public health because seropositive pigs can harbor tissue cysts in their meat, thereby representing a zoonotic risk for consumers of raw or undercooked porcine meat or its products.

Detection of Mycoplasma species at various anatomical sites of dogs from different types of kennels.

This work aimed to detect Mycoplasma cynos, M. canis, M. edwardii, and M. molare in different types of kennels, in addition to evaluating their distribution in different colonization sites. The dogs belonged to different kennels from armed forces (n = 3), shelters (n = 3), and commercial purposes (n = 2). Samples of the oropharynx, genital mucosa, and ear canal were collected from each dog (n = 98), totaling 294 samples. Aliquots were submitted to isolation and the samples confirmed as Mycoplasma spp. were subjected to conventional PCR for M. canis and multiplex PCR for M. edwardii, M. molare, and M. cynos detection. Of the 98 dogs studied, 63.3% (62) were positive in at least one anatomical site evaluated for Mycoplasma spp. Among the 111 anatomical sites positive for Mycoplasma spp., M. canis, M. edwardii, and M. molare were detected in 29.7% (33/111), 40.5% (45/111), and 2.70% (3/111), respectively. No animal was positive for M. cynos.

Toxoplasma gondii infection in pig intended for human consumption: seroprevalence, risk factors and influence of biosecurity measures / Infecção por Toxoplasma gondii em suínos destinados ao consumo humano: soroprevalência, fatores de risco e influência de medidas de biosseguridade

Abstract A serologic and epidemiologic study was carried out in order to determinate herd and animal seroprevalence and associated factors for Toxoplasma gondii in commercial pigs from Espírito Santo state, Brazil. Blood samples were collected from 416 pigs from 55 producer farms in 27 municipalities. An indirect immunofluorescent assay (IFA) was performed to estimate the seroprevalence of T. gondii and identify the associated risk factors using a questionnaire. The T. gondii antibody prevalence rate in commercial swine herds was 15.4% (64/416) using a cutoff of 1:64. The seropositivity for T. gondii was related to the presence of cats, water origin and age of swine in the increase of seroprevalence, and the existence of internal isolation fences and use of composting chambers as protective factors. To the best of our knowledge, this is the first study to report anti- T. gondii antibodies in the serum of pigs in the state of Espírito Santo, Brazil. This finding is important to public health because seropositive pigs can harbor tissue cysts in their meat, thereby representing a zoonotic risk for consumers of raw or undercooked porcine meat or its products.

Resumo Um estudo sorológico e epidemiológico foi realizado com o objetivo de determinar a soroprevalência nas granjas e animais e também em fatores associados para Toxoplasma gondii em suínos comerciais do estado do Espírito Santo, Brasil. Foram coletadas amostras de sangue de 416 suínos de 55 granjas produtoras de 27 municípios. A reação de imunofluorescência indireta (RIFI) foi realizada com o uso de questionário, para estimar a soroprevalência do T. gondii e identificar os fatores de risco associados. A taxa de prevalência de anticorpos contra T. gondii, nos rebanhos comerciais de suínos, foi de 15,4% (64/416) com ponto de corte de 1:64. A soropositividade para T. gondii foi relacionada à presença de gatos, origem da água, a idade dos suínos no aumento da soroprevalência, a existência de cercas de isolamento interno e uso de câmaras de compostagem como fatores de proteção. De acordo com este estudo, ele é o primeiro a relatar anticorpos anti-T. gondii no soro de suínos no estado do Espírito Santo, Brasil. Este achado é importante para a saúde pública, pois suínos soropositivos podem abrigar cistos teciduais em suas carnes, representando um risco zoonótico para os consumidores de carne suína crua ou mal cozida ou seus produtos.

Molecular markers associated with antimicrobial resistance and genotypes of Campylobacter jejuni and Campylobacter coli isolated from broiler and swine flocks in southeast Brazil.

This study aimed to identify molecular markers associated with antimicrobial resistance and genotype isolates of Campylobacter spp. from broiler and swine flocks due to its importance to one-health. C. jejuni (n=27) and C. coli (n = 35) strains were screened for the antimicrobial genetic markers C257T in gyrA, A2074C and A2075G in 23S rRNA, CmeABC, ermB, tetO and blaOXA61 by PCR. Fifteen strains had SVR-flaA and porA genes sequenced to evaluate their genetic diversity. Among C. jejuni strains 62.96% had C257T mutation and only one strain had A2075G mutation. CmeA, cmeB, cmeC, tetO and blaOXA61 were detected respectively in 92.59%, 100%, 100%, 85.19%, 85.19% of the strains. All C. coli had C257T mutation; 48.75% had A2075G and cmeA, cmeB, cmeC, tetO, blaOXA61 were detected in 8.57%, 94.29%, 91.43%, 91.43%, 80%, respectively. Twelve porA and SVR-flaA alleles were detected, with a Simpson index of diversity value of 0.962.

First molecular detection of Spiroplasma spp. in ticks from horses in Brazil.

The class Mollicutes comprises microorganisms that lack a cell wall, highly dependent on their host to survive. Within Mollicutes, the genus Spiroplasma comprises motile helical microorganisms associated with various insects and other arthropods. This study aimed to detect and characterize Mollicutes microorganisms in ticks of different species of veterinary importance, using molecular techniques. These ticks were collected from dogs, cats, cattle, and horses from Rio de Janeiro's metropolitan regions. They were morphologically classified and pooled according to their species for subsequent DNA extraction. These samples were tested by PCR using class Mollicutes-specific primers (16S rRNA) and positive amplicons were sequenced. The obtained DNA sequences were compared with other Mollicutes sequences deposited in GenBank. We found that four out of 745 (0.54%) of the tick pools were positive for members of the class Mollicutes, identified as Spiroplasma spp.; of the positive pools, one comprised Amblyomma sculptum adults and three comprised Dermacentor nitens nymphs. The present study describes Spiroplasma spp. in ticks in Brazil for the first time. Nevertheless, due to few reports on these microorganisms, further studies on epidemiology, virulence, and pathogenicity are needed.

Risk factors associated with intramammary colonization with Mollicutes in dairy cattle from Southeast Brazil / Fatores de risco associados à colonização intramamária por Mollicutes em bovinos leiteiros do Sudeste brasileiro

ABSTRACT: Bacteria of Mollicutes Class are associated with intramammary infection and decrease in milk production. This study investigated the occurrence of Mollicutes and elucidated their risk factors in dairy herds from Southeast Brazil. For this, milk samples from 387 lactation cows from Minas Gerais, Rio de Janeiro and São Paulo States were subjected to the polymerase chain reaction (PCR) to detect Mollicutes. Species of Mycoplasma were investigated in Mollicutes positive samples by PCR, including Mycoplasma bovis, M. alkalescens, M. bovigenitalium, M. bovirhinis, M. arginini and A. laidlawii. An epidemiological questionnaire was applied to collect data on possible risk factors, which were assessed using Pearson's Chi-square test followed by odds ratio (P≤0.05). Mollicutes were reported in 21% (4/19) of the herds and 4% (16/387) of the animals, while 1% (5/387) were positive for M. bovis and 3% (11/387) for M. arginini. All samples were negative to the other agents. Herds with more than 150 animals [OR=3.51 (95% CI 1.11-11.08)], manual milking [OR=9.97 (95% CI 2.80-35.49)] and not-milking animals with mastitis last [OR=6.54 (95% CI 1.92-22.29)] were risk factors. The presence of these conditions may favor intramammary infection by Mollicutes in dairy herds from Southeast Brazil. This is the first report of M. bovis in Rio de Janeiro and M. arginini in the studied states.

RESUMO: Bactérias da Classe Mollicutes estão associadas à infecção intramamária e diminuição da produção leiteira. O objetivo deste estudo foi investigar a ocorrência de Mollicutes e elucidar seus fatores de risco em rebanhos leiteiros do sudeste brasileiro. Para isso, amostras de leite de 387 vacas em lactação dos estados de Minas Gerais, Rio de Janeiro e São Paulo foram submetidas à reação em cadeia da polimerase (PCR) para detectar Mollicutes. Espécies de Mycoplasma foram investigadas nas amostras positivas por PCR, incluindo Mycoplasma bovis, M. alkalescens, M. bovigenitalium, M. bovirhinis, M. arginini e A. laidlawii. Foi aplicado um questionário epidemiológico para a coleta de dados sobre possíveis fatores de risco, que foram avaliados pelo teste de Qui-Quadrado de Pearson seguido de odds ratio (P≤0.05). Mollicutes foram detectados em 21% (4/19) dos rebanhos e 4% (16/387) dos animais, enquanto 1% (5/387) destes foram positivos para m. bovis, 3% (11/387) para m. arginini, sendo todas as amostras negativas para os demais agentes. Rebanhos com mais de 150 animais [OR=3,51 (95% IC 1,11-11,08)], ordenha manual [OR=9,97 (95% IC 2,80-35,49)] e ausência de linha de ordenha [OR=6,54 (95% IC 1,92-22,29)] foram considerados fatores de risco. A presença dessas condições pode favorecer a infecção intramamária por Mollicutes em rebanhos leiteiros no sudeste do Brasil. Este é o primeiro relato de M. bovis no Rio de Janeiro e M. arginini nos estados estudados.

Mycoplasma spp. in captive snakes (Boa constrictor and Bothrops atrox) from Brazil / Mycoplasma spp. em cobras de cativeiro (Boa constrictor e Bothrops atrox) do Brasil

ABSTRACT: Although rare, mycoplasmas are included among the causes of respiratory diseases in reptiles and, in the order Squamata, three reports of these microorganisms causing diseases in pythons have already been reported. This study aimed to evaluate the occurrence of Mycoplasma species in captive snakes. A total of 26 snakes of the families Pythonidae (13), Boidae (7), Viperidae (5) and Colubridae (1) from RioZoo, Brazil, were evaluated. Animals were examined to determine clinical signs consistent with any infectious disease. Tracheal swab samples from snakes were collected in Frey medium and analyzed for the presence of Mycoplasma spp.by isolation and a genus-specific PCR. DNA sequencing analyses of six positive samples by PCR were carried out to identify the species. Using isolation 19.23% (5/26) was positive, while 65.38% (17/26) of the animals were positive by PCR. Based on the analyses of the six sequences obtained, there was similarity with a Mycoplasma spp. previously described in a phyton and, M. agassizii and M. testudineum reported in chelonians. This is the first report of Mycoplasma spp. in animals of the families Boidae and Viperidae. Mycoplasma spp. were detected in snakes with and without clinical signs. The mycoplasmas reported resented identity (range, 95% to 100%) to others already described in reptiles. There was no relationship between the presence of Mycoplasma spp. and clinical signs.

RESUMO: Embora raros, os micoplasmas estão incluídos entre as causas de doenças respiratórias em répteis e, na ordem Squamata, já foram realizados três relatos destes microrganismos causando doença em pítons. Este estudo teve como objetivo avaliar a ocorrência de espécies de Mycoplasma em serpentes em cativeiro. Foram avaliadas 26 serpentes das famílias Pythonidae (13), Boidae (7), Viperidae (5) e Colubridae (1) do RioZoo, Brasil. Os animais foram examinados para determinar sinais clínicos consistentes com qualquer doença infecciosa. Amostras de swab traqueal de cobras foram coletadas em meio Frey e analisadas por isolamento microbiológico e pela técnica da PCR para identificar Mycoplasma spp. As amostras positivas para o gênero Mycoplasma spp. foram submetidas ao sequenciamento genético para identificação das espécies. No isolamento, 19,23% (5/26) foram positivos, enquanto 65,38% (17/26) dos animais foram positivos por PCR. Com base nas análises das seis sequências obtidas, houve similaridade com o Mycoplasma spp. descrito anteriormente em um píton e M. agassizii e M. testudineum encontrados em quelônios. Este é o primeiro relato de Mycoplasma spp. em animais das famílias Boidae e Viperidae. Mycoplasma spp. foi detectado em serpentes com e sem sinais clínicos. Os micoplasmas encontrados apresentaram semelhança genética com outros já descritos em répteis. Não houve relação entre a presença de Mycoplasma spp. e sinais clínicos.

Eustrongylides sp. (Nematoda, Dioctophymatoidea) parasitizing Hoplias malabaricus (Actinopterygii: Erythrinidae) collected from the state of Rio de Janeiro, Brazil / Eustrongylides sp. (Nematoda, Dioctophymatoidea) parasitando Hoplias malabaricus (Actinopterygii: Erythrinidae) coletados do Estado do Rio de Janeiro, Brasil

Abstract Hoplias malabaricus (Characiformes, Erythrinidae), trahira, is a neotropical freshwater fish of economic and public health significance. A total of 45 specimens of H. malabaricus commercialized in the municipality of Magé, state of Rio de Janeiro, Brazil, were acquired between April 2016 and April 2018 to investigate the presence of nematode larvae. Twenty of the fish were found parasitized by 347 fourth-stage nematode larvae identified taxonomically as Eustrongylides sp. using morphological, morphometric and molecular data. The parasitic indices were: prevalence 44.44%, mean intensity 17.35, mean abundance 7.71, and range of infection 2-40. Infection sites were musculature, mesentery, abdominal cavity, and serosa of intestine, stomach and liver. This is the first report of Eustrongylides sp. larvae parasitizing H. malabaricus in the state of Rio de Janeiro.

Resumo Hoplias malabaricus (Characiformes, Erythrinidae), traíra, é um peixe neotropical de água doce que tem significante impacto na economia e saúde pública. De abril de 2016 a abril de 2018, foram adquiridos 45 espécimes de H. malabaricus comercializados no município de Magé, Estado do Rio de Janeiro, Brasil. Os peixes foram necropsiados e filetados para investigação da presença de larvas de nematoides. Vinte dos peixes coletados estavam parasitados por 347 larvas de nematoides, identificadas taxonomicamente como larvas de quarto estágio de Eustrongylides sp. usando-se dados morfológicos, morfométricos e moleculares, apresentando os seguintes valores: prevalência de 44,44%, intensidade média de 17,35, abundância média de 7,71, e amplitude de variação da infecção de 2-40. Os sítios de infecção foram musculatura, mesentério, cavidade abdominal e serosas do intestino, estômago e fígado. Este é o primeiro registro de larvas de Eustrongylides sp. parasitando H. malabaricus no Estado do Rio de Janeiro.

Eustrongylides sp. (Nematoda, Dioctophymatoidea) parasitizing Hoplias malabaricus (Actinopterygii: Erythrinidae) collected from the state of Rio de Janeiro, Brazil.

Hoplias malabaricus (Characiformes, Erythrinidae), trahira, is a neotropical freshwater fish of economic and public health significance. A total of 45 specimens of H. malabaricus commercialized in the municipality of Magé, state of Rio de Janeiro, Brazil, were acquired between April 2016 and April 2018 to investigate the presence of nematode larvae. Twenty of the fish were found parasitized by 347 fourth-stage nematode larvae identified taxonomically as Eustrongylides sp. using morphological, morphometric and molecular data. The parasitic indices were: prevalence 44.44%, mean intensity 17.35, mean abundance 7.71, and range of infection 2-40. Infection sites were musculature, mesentery, abdominal cavity, and serosa of intestine, stomach and liver. This is the first report of Eustrongylides sp. larvae parasitizing H. malabaricus in the state of Rio de Janeiro.

Callitetrarhynchus gracilis (Rudolphi, 1819) Pintner, 1931 (Cestoda: Trypanorhyncha) parasitizing the musculature of Sardinella brasiliensis (Steindachner, 1879) (Actinopterygii) off the coast of the state of Rio de Janeiro, Brazil.

Cestodes of the order Trypanorhyncha can frequently be found infecting the muscles of several marine fish species, and lead to a repugnant aspect of the fish and rejection by consumers. The Brazilian sardinella, Sardinella brasiliensis, occurs from the Gulf of Mexico and the Caribbean to northern Uruguay. In southeastern Brazil, fishing for the species is very intensive since it generates significant revenue as one of the most commonly consumed fish and an important element of the canned fish industry. The aims of the present study were to identify and report the occurrence of tiny cestodes (3 mm-6.6 mm) in the musculature of Brazilian sardinella that were purchased in the São Pedro fish market in the municipality of Niterói, state of Rio de Janeiro, Brazil. From October 2013 to November 2016, 75 specimens of S. brasiliensis were investigated. The trypanorhynch cestodes encountered were identified as plerocerci of Callitetrarhynchus gracilis using morphological, morphometric and molecular data. Parasitic indices were calculated, and the cestodes infection of the musculature of Brazilian sardinella had the following values: prevalence, 40%; mean intensity, 3.47; mean abundance, 1.39; and range of infection, 1-18, specimens. The presence of this trypanorhynch cestode in the musculature of Brazilian sardinella is an important indicator of fish hygiene.

Ações de controle populacional de cães não domiciliados realizadas pelo poder público em municípios do Rio de Janeiro, Brasil (2012-2013) / Populational control actions for stray dogs carried out by the public authorities in municipalities of Rio de Janeiro, Brazil (2012-2013)

A grande densidade populacional de cães não domiciliados é uma realidade nos municípios brasileiros, trazendo problemas à ordem urbana e à saúde coletiva. Este trabalho descreve as ações desenvolvidas pelos órgãos de saúde dos municípios do Rio de Janeiro, no período 2012-2013, visando ao controle populacional de cães não domiciliados. Após a seleção de uma amostra estatisticamente significativa, composta por 47 municípios, houve a aplicação de questionário aos responsáveis pelos serviços de controle de zoonoses desses municípios. Somente 46,8% (n=22) realizavam alguma ação relacionada ao controle populacional. As principais ações desenvolvidas eram as atividades educativas sobre guarda responsável, a esterilização gratuita e os projetos de adoção, mesmo assim presentes em menos de 30% dos municípios. Já o recolhimento sistemático com posterior eutanásia não é mais uma prática frequente no Estado do Rio de Janeiro, não tendo sido relatado por nenhum município avaliado. De modo geral as poucas ações visando ao controle populacional de cães não domiciliados desenvolvidas pelos municípios avaliados, em todas as Regiões de Saúde, eram fragmentadas e não seguiam um protocolo de atuação com ações integradas.

Performance, serologic and tracheal responses of laying hens exposed to F strain of Mycoplasma gallisepticum / Desempenho, sorologia e respostas traqueias de galinhas poedeiras expostas a cepa F de Mycoplasma gallisepticum

The F strain of Mycoplasma gallisepticum (MG-F) protects chickens against mycoplasma infections, in which monitoring is made by serology and histopathology of trachea. This trial used 90 chickens, being 30 unvaccinated (G1 group), 30 eye-drop vaccinated at 8 weeks of age with MG-F (Ceva Animal Health, São Paulo, SP, Brazil) (G2), and 30 immunized at 8 and 11 weeks of age (G3). Samples were obtained from chickens on the 8, 12, 15, 18, 20 and 24th weeks of age for the enzyme-linked immunosorbent assay (ELISA) test. Tracheal fragments were collected after necropsies on the 15 and 24th weeks of age. Up to 12 weeks, the ELISA reactions in optical density (OD) were 0.165 (G1), 0.151 (G2) and 0.151(G3), all below 0.20 and with no significant difference among groups (p > 0.05). After the 15th week, the ELISA reactions rose, yielding the following group averages by collecting dates: G1 (0.18, 0.19, 0.18, and 0.16), G2 (0.36, 0.49, 0.47, and 0.44) and G3 (0.41, 0.52, 0.59, 0.60), being the means in G2 and G3 not significantly different between than, but significantly different from G1. The initial weight (592.71, 621.33, and 594.40), the final weight (1,932.58, 1,987.59, and 1,875.20) and the weekly weight gain (11.65, 11.90, and 11.14) were not significantly different among groups. At necropsy the gross tracheal score means by group and dates were: 15th week (0.25, 0.61, and 0.54) and 24th week (0.54, 0.58, and 0.67), being these difference not significantly (p > 0.05). On microscopy, the tracheal score averages by groups G1, G2 and G3, respectively, were: 15th week (0.25, 0.32, and 0.47) and 24th week (0.07, 0.75, and 0.08). G2 yielded higher score average than G1 and G3 on the 24th week. Higher tracheal changes for G2 and G3 as compared to G1 could be ascribed to MG-F infection. There were no evident prejudicial effects on live weight, weight gain and tissue changes by applying one or two vaccination doses.(AU)

A cepa F de Mycoplasma gallisepticum (MG-F) protege as galinhas de micoplasmose, e sua monitorização é feita por sorologia e histopatologia de traqueia. Este estudo utilizou 90 frangos, sendo 30 não vacinados (grupo G1); 30 vacinados via gota ocular a 8 semanas de idade com MG-F (Ceva Saúde Animal, São Paulo, SP, Brasil) (G2); e 30 imunizados em 8 e 11 semanas de idade (G3). As amostras foram obtidas nas 8ª, 12ª, 15ª, 18ª, 20ª e 24ª semanas para ensaio de imunoabsorção enzimática (ELISA). Fragmentos traqueais foram coletados após necropsias nas 15ª e 24ª semanas. Até a 12ª semana, as reações de ELISA em densidade óptica (DO) foram 0,165 (G1), 0,151 (G2) e 0,151 (G3), todas abaixo de 0,20, e não houve diferença significativa entre os grupos (p > 0,05). Após a 15ª semana , a reação de ELISA subiu, produzindo as seguintes médias dos grupos por datas de coleta: G1 (0,18, 0,19, 0,18 e 0,16), G2 (0,36, 0,49, 0,47 e 0,44) e G3 (0,41, 0,52, 0,59, 0,60), sendo as médias de G2 e G3 não significativamente diferentes entre si, mas significativamente diferentes da de G1. O peso inicial (592,71, 621,33, 594,40), o peso final (1.932,58, 1.987,59, 1.875,20) e o ganho de peso semanal (11,65, 11,90, 11,14) não foram significativamente diferentes entre os grupos. Na necropsia, as médias do escore da macroscopia de traqueia por grupo e data foram: 15ª semana (0,25, 0,61 e 0,54) e 24ª semana (0,54, 0,58 e 0,67), e não se apresentou diferença significativa (p > 0,05). Na microscopia, a média de escores de traqueia por grupos G1, G2 e G3, respectivamente, foram: 15ª semana (0,25, 0,32 e 0,47) e 24ª semana (0,07, 0,75 e 0,08). G2 apresentou maior média de escore do que G1 e G3 na 24ª semana. Alterações traqueais mais elevadas para G2 e G3 em relação a G1 poderiam ser atribuídas à vacinação por MG-F. Não houve efeitos prejudiciais evidentes no peso vivo nem no ganho de peso, tampouco alterações teciduais na aplicação de uma ou duas doses de vacinação.(AU)

Chlamydia psittaci in captive blue-and-gold macaws (Ara ararauna) in a triage center of wild animals in Brazil / Chlamydia psittaci em araras-canindé (Ara ararauna) cativas em um Centro de Triagem de Animais Silvestres no Brasil

Chlamydia psittaci infection stands out due to its zoonotic potential. It was detected in several orders, being Psittaciformes its main reservoir. Our objective was to investigate the occurrence of C. psittaci infection by clinical and environmental examination, as well as agent detection by Polymerase Chain Reaction (PCR) from choanal and cloacal swabs in 46 blue-and-gold macaws (Ara ararauna) allocated at the Triage Center of Wild Animals, Brazilian Institute of Environment and Renewable Natural Resources, Rio de Janeiro state, Brazil. The frequency of positive detection of the bacteria found was 50% (23/46) by cloacal swab. In this percentual, 26.09% (12/23) were also positive by choanal swab, being the probability of detection 2.83 times higher for cloacal swabs when compared to choanal ones (p<0.05). There was no association between body condition, temperature variations and PCR positivity for C. psittaci, being 34.78% of positive individuals asymptomatic to this disease. Therefore, our findings show agent's dispersion at the squad and high frequency of asymptomatic birds. Considering the possibility of contagion to workers and free living birds, the quarantine in wildlife conservation and support centers, as well as the care with hygiene and individual protection for humans who deal with the animals, should be strictly followed even in the absence of clinical signs.

Infecções por Chlamydia psittaci destacam-se devido ao seu potencial zoonótico. Tal agente foi detectado em diversas ordens, sendo Psittaciformes seu principal reservatório. Objetivando-se demonstrar a ocorrência de tal processo infeccioso, procedeu-se investigação clínica e ambiental, associada à detecção do agente etiológico por Reação em Cadeia da Polimerase (PCR), a partir de swabs de coana e cloaca de 46 araras-canindés (Ara ararauna) pertencentes ao Centro de Triagem de Animais Silvestres do Instituto Brasileiro de Meio Ambiente e Recursos Renováveis, Rio de Janeiro, Brasil. Obteve-se uma frequência de detecção de 50% (23/46) a partir de swabs de cloaca. Entre esses, 26.09% (12/23) foram também positivos por meio de swab de coana, sendo a probabilidade de detecção 2,83 vezes maior a partir de swabs de cloaca que de coana (p<0.05). Não houve associação entre condição corporal, variação de temperatura e detecção do agente por PCR, sendo 34,78% dos indivíduos positivos assintomáticos. Portanto, nossos resultados demonstram a dispersão do agente no plantel e alta frequência de assintomáticos. Devido à possibilidade de contágio para trabalhadores e aves de vida livre, a quarentena nos centros de suporte e conservação da vida silvestre, assim como os cuidados com higiene e proteção individual para com os trabalhadores que lidam com os animais devem ser seguidas rigidamente, mesmo na ausência de sinais ou sintomas nas aves

Validation of PCR method for mycoplasma detection in the Yellow Fever-vaccine quality control / Validação de método de PCR para detecção de micoplasmasno controle de qualidade da vacina de Febre Amarela

Among the vaccines produced by Bio-Manguinhos, a major centre for manufacturingthe immunobiological products in Latin America, stands out the yellow fever (YF) vaccine.To guarantee the excellence and safety of the YF vaccine, the quality control tests has to be performedthroughout its production. The World Health Organization (WHO) demands the producersto guarantee the absence of Mycoplasma orale, M. pneumoniae, M. gallisepticum and M. synoviaein the biological products. Mycoplasma is a fastidious microorganism, requiring about 35 daysfor attaining the conclusive culturing test. In this study PCR methods were selected for amplifying16S rRNA gene fragments for detecting mycoplasma in the intermediate products of YF vaccine.This standardized methodology was specific and sensitive to detect the low concentrations of mycoplasma in spiked intermediary vaccine products; and the absence of unspecific amplification was also demonstrated. The detection rates ranged from 3.1 to 12.5 colony formingunits and showed 100 % of sensitivity and specificity in the tested samples. The PCR protocol for detecting mycoplasmal DNA in YF vaccine was validated by analysing 286 samples. Bio-Manguinhos produces annually 10,000,000 YF vaccine doses, and this method has been successfully employed, complementing the traditional approach in the mycoplasma detection since 2008.

Dentre as vacinas produzidas por Bio-Manguinhos, um importante centro de produção deimunobiológicos da América Latina, destaca-se a vacina de febre amarela (FA) que é produzidaem ovos embrionados. Para garantir a excelência e a segurança da vacina, testes de controlede qualidade são realizados durante a produção. A Organização Mundial de Saúde (OMS) exigedos produtores a ausência de Mycoplasma orale, M. pneumoniae, M. gallisepticum e M. synoviaeem produtos biológicos. Micoplasmas são micro-organismos fastidiosos, sendo necessários35 dias para que os testes de cultura sejam conclusivos. Neste estudo foram selecionadosmétodos de amplificação de fragmentos do gene 16S rRNA para detecção de micoplasmas emprodutos intermediários da vacina de FA. Esta metodologia padronizada foi capaz de detectarbaixas concentrações de micoplasmas nos produtos intermediários e a ausência de amplificaçãoinespecífica foi demonstrada. O limite de detecção variou entre 3,1 e 12,5 unidades formadorasde colônia; e nas amostras testadas a sensibilidade e a especificidade foram de 100 %. O protocolode PCR para detecção de micoplasmas na vacina foi validado pela análise de 286 amostras.Bio-Manguinhos produz 10.000.000 doses de vacina de febre amarela por ano e, desde 2008,este método tem sido empregado com sucesso, complementando-se a abordagem tradicional.

Validation of PCR method for mycoplasma detection in the Yellow Fever-vaccine quality control / Validação de método de PCR para detecção de micoplasmas no controle de qualidade da vacina de Febre Amarela

Among the vaccines produced by Bio-Manguinhos, a major centre for manufacturing the immunobiological products in Latin America, stands out the yellow fever (YF) vaccine. To guarantee the excellence and safety of the YF vaccine, the quality control tests has to be performed throughout its production. The World Health Organization (WHO) demands the producers to guarantee the absence of Mycoplasma orale, M. pneumoniae, M. gallisepticum and M. synoviae in the biological products. Mycoplasma is a fastidious microorganism, requiring about 35 days for attaining the conclusive culturing test. In this study PCR methods were selected for amplifying 16S rRNA gene fragments for detecting mycoplasma in the intermediate products of YF vaccine. This standardized methodology was specific and sensitive to detect the low concentrations of mycoplasma in spiked intermediary vaccine products; and the absence of unspecific amplification was also demonstrated. The detection rates ranged from 3.1 to 12.5 colony forming units and showed 100 % of sensitivity and specificity in the tested samples. The PCR protocol for detecting mycoplasmal DNA in YF vaccine was validated by analysing 286 samples. Bio-Manguinhos produces annually 10,000,000 YF vaccine doses, and this method has been successfully employed, complementing the traditional approach in the mycoplasma detection since 2008.

Dentre as vacinas produzidas por Bio-Manguinhos, um importante centro de produção de imunobiológicos da América Latina, destaca-se a vacina de febre amarela (FA) que é produzida em ovos embrionados. Para garantir a excelência e a segurança da vacina, testes de controle de qualidade são realizados durante a produção. A Organização Mundial de Saúde (OMS) exige dos produtores a ausência de Mycoplasma orale, M. pneumoniae, M. gallisepticum e M. synoviae em produtos biológicos. Micoplasmas são micro-organismos fastidiosos, sendo necessários 35 dias para que os testes de cultura sejam conclusivos. Neste estudo foram selecionados métodos de amplificação de fragmentos do gene 16S rRNA para detecção de micoplasmas em produtos intermediários da vacina de FA. Esta metodologia padronizada foi capaz de detectar baixas concentrações de micoplasmas nos produtos intermediários e a ausência de amplificação inespecífica foi demonstrada. O limite de detecção variou entre 3,1 e 12,5 unidades formadoras de colônia; e nas amostras testadas a sensibilidade e a especificidade foram de 100 %. O protocolo de PCR para detecção de micoplasmas na vacina foi validado pela análise de 286 amostras. Bio-Manguinhos produz 10.000.000 doses de vacina de febre amarela por ano e, desde 2008, este método tem sido empregado com sucesso, complementando-se a abordagem tradicional.

[Rabies in the State of Rio de Janeiro, Brazil: analysis of surveillance and control actions in the municipal field]. / Raiva no Estado do Rio de Janeiro, Brasil: análise das ações de vigilância e controle no âmbito municipal.

Rabies is an anthropozoonosis characterized by acute viral encephalitis with a lethality rate close to 100%, and it has undergone an epidemiologic transition in which the cycle involving chiroptera is increasing in importance. The scope of this research sought to analyze the rabies surveillance and control actions carried out in municipalities in the State of Rio de Janeiro. Questionnaires were distributed to a representative sample of zoonosis control service managers proportionately calculated in accordance with the Health Regions, according to the State Regionalization Guidance Plan. The data gathered was recorded and analyzed using descriptive statistical techniques. Based on the results attained, the conclusion reached is that the rabies surveillance and control actions were being unsatisfactorily conducted, especially for items related to the monitoring of vampire bat colonies, viral circulation surveillance, notification and monitoring of suspect or aggressive animals, quantification of dog population and population control of stray dogs. The surveillance and control of rabies was being neglected, and was not a priority in the health services in the municipalities evaluated.

Raiva no Estado do Rio de Janeiro, Brasil: análise das ações de vigilância e controle no âmbito municipal / Rabies in the State of Rio de Janeiro, Brazil: analysis of surveillance and control actions in the municipal field

A raiva é uma antropozoonose caracterizada por encefalite viral aguda, com letalidade próxima de 100%, e que vem passando por uma transição epidemiológica na qual o ciclo envolvendo quirópteros vem crescendo em importância. O objetivo da presente pesquisa foi analisar as ações de vigilância e controle da raiva desenvolvidas em municípios do Estado do Rio de Janeiro. Foram aplicados questionários a uma amostra representativa de gestores dos serviços de controle de zoonoses, proporcionalmente calculada em função das Regiões de Saúde, de acordo com o Plano Diretor de Regionalização do Estado. Os dados foram tabulados e trabalhados com técnicas de estatística descritiva. Com base nos resultados encontrados pode-se concluir que as ações de vigilância e controle da raiva estavam sendo desenvolvidas de maneira insatisfatória, principalmente nos itens monitoramento das colônias de morcegos hematófagos, vigilância da circulação viral, notificação e acompanhamento de animais suspeitos ou agressores, quantificação da população canina e controle populacional de cães não domiciliados. A vigilância e o controle da raiva estavam sendo negligenciados e não eram uma prioridade dos serviços de saúde dos municípios avaliados.

Rabies is an anthropozoonosis characterized by acute viral encephalitis with a lethality rate close to 100%, and it has undergone an epidemiologic transition in which the cycle involving chiroptera is increasing in importance. The scope of this research sought to analyze the rabies surveillance and control actions carried out in municipalities in the State of Rio de Janeiro. Questionnaires were distributed to a representative sample of zoonosis control service managers proportionately calculated in accordance with the Health Regions, according to the State Regionalization Guidance Plan. The data gathered was recorded and analyzed using descriptive statistical techniques. Based on the results attained, the conclusion reached is that the rabies surveillance and control actions were being unsatisfactorily conducted, especially for items related to the monitoring of vampire bat colonies, viral circulation surveillance, notification and monitoring of suspect or aggressive animals, quantification of dog population and population control of stray dogs. The surveillance and control of rabies was being neglected, and was not a priority in the health services in the municipalities evaluated.

Pré-resfriamento na redução de coliformes em carcaças de frango de corte / Immersion chilling in fecal coliforms count reduction on broiler carcasses

O objetivo deste trabalho foi comparar a análise individual (plano de duas classes) e a análise por lotes (plano de três classes), priorizado pelo plano amostragem oficial da ANVISA, na influência do pré-resfriamento de carcaças de frangos de corte na redução da contaminação por coliformes termotolerantes. Foram analisadas 240 carcaças de frangos de corte, sendo coletadas 120 amostras antes e 120 após a etapa de pré-resfriamento, para quantificação de coliformes termotolerantes pela técnica de contagem em placas. As médias das contagens obtidas das carcaças coletadas antes e após o pré-resfriamento foram diferentes, com uma redução média de 0,99log10 UFC g-1 de coliformes termotolerantes. Na interpretação dos resultados obtidos antes do pré-resfriamento pela análise individual, 16,7% (20/120) das carcaças foram classificadas como inaceitáveis, enquanto, pela análise por lotes, foram 37,5% (45/120). Houve associação entre a aceitabilidade dos lotes e a passagem pelo chiller com um valor de Odds Ratio de 35,48. Ficou demonstrada a importância da utilização do plano de análise por lotes e da etapa de pré-resfriamento no processo de produção, sendo decisivos para a aceitação dos lotes de carcaças de aves para comercialização pelos parâmetros vigentes na legislação nacional.

The objective of this study was to assess the influence of immersion chilling on broiler carcasses contamination by fecal coliforms counting in a poultry slaughterhouse under Federal Sanitary Inspection by individual and three class plan interpretation of the results. Two hundred and forty broiler carcasses, being collected 120 before and 120 after passage thought chillers, were analyzed for coliform counting by plate count technique. The averages of the counts obtained from all carcasses collected before and after immersion chilling were different, with an average reduction of 0.99log10 CFU g-1 of fecal coliforms. Regarding individual results (two class plan) before chilling, 16.7% (20/120) of the carcasses were classified as unacceptable, against 37.5% (45/120) regarding lots results (three class plan). There was a strong association between the acceptability of lots and immersion chilling with an Odds Ratio value of 35.48. It was demonstrated the importance of the official sampling plan and immersion chilling in the production process, being decisive for the acceptance of the chicken carcass lots by the national legislation standards.

Detection of Ureaplasma spp. in semen samples from sheep in Brazil

A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples.

Cross-sectional study of serum reactivity to Anisakis simplex in healthy adults in Niterói, Brazil.

Although the incidence of anisakiasis is rising worldwide, its frequency is still unknown in Brazil. The aim of this study was to verify immunoreactivity to Anisakis simplex antigens in healthy adults and determine its possible relationship with frequency of fish consumption and allergy symptoms. A prospective cross-sectional study was carried out with 67 volunteers recruited from a military facility in Niterói, Brazil. The subjects completed a structured questionnaire and serum titers of specific anti-Anisakis IgE and IgG antibodies were measured. The association between frequency of fish intake and IgE reactivity was evaluated by Fisher's exact test. Almost all subjects (97.0%, 65/67) that consumed seafood; 64.6% (42/65) ate fish at least once weekly. Of all seafood consumers, 56.9% (37/65) reported allergy symptoms, being gut allergies most often cited (35.5%). IgE seroreactivity to Anisakis simplex was found in 20.9% of subjects (14/67), with 13.4% (9/67) reacting exclusively to somatic antigen, 3.0% (2/67) exclusively to excretory/secretory antigens and 4.5% (3/67) to both antigens. There was a significant association between frequency of fish consumption and positive serology (p = 0.019). An immunoblot assay for Anisakis antigens showed different positive bands for IgG. The direct relationship between ELISA reactivity and frequency of fish intake and absence of association with allergy symptoms suggests previous contact with Anisakis simplex antigens.