Hematologic reference values of Vinaceous-breasted Amazon (Amazona vinacea) / Valores hematológicos de referência em Papagaio-de-Peito-Roxo (Amazona vinacea)

Avian hematologic reference intervals are useful tools to evaluate body homeostasis and diagnose diseases. However, there are few species-specific reference intervals published. The present study reports Vinaceous-breasted Amazon ( Amazona vinacea ) hematologic reference values obtained during the health status evaluation of release candidates as part of this species reintroduction efforts at the Araucárias National Park. Parameters reported are erythrogram (erythrocytes, hemoglobin, packed cell volume, mean cell volume, mean corpuscular hemoglobin concentration and mean corpuscular hemoglobin), Red Cell Distribution Width (RDW), white cells total and differential (heterophiles, lymphocytes, basophils, eosinophils and monocytes), thrombocytes and total plasma protein. For the first time results on RDW and thrombocytes were described and a larger sample size were provided for all parameters analyzed. Intervals demonstrated in the present study showed significant differences from those considered normal in other parrot species and consequently have contributed to bring valuable information to base actions for the conservation of this endangered species of great biological value.(AU)

Os valores de intervalos de referência para parâmetros hematológicos aviários são ferramentas úteis para avaliação da homeostase corporal e diagnóstico de doenças. No entanto, são escassos os relatos de intervalos de referência espécie-específico, o que aumenta a probabilidade de um erro na interpretação de dados laboratoriais. Dessa forma, o presente estudo relata valores hematológicos de referência específicos para papagaio-de-peito-roxo ( Amazona vinacea ). Esses valores foram obtidos e calculados durante um projeto de reintrodução como importante e adequada forma para avaliar o estado de saúde de candidatos à soltura. Os analíticos observados foram: eritrograma (eritrócitos, hemoglobina, hematócrito, volume celular, concentração de hemoglobina corpuscular e hemoglobina corpuscular média), distribuição das células vermelhas (RDW), leucograma (basófilos, eosinofilis mielócitos, metamielócitos, heterófilos, linfócitos e monócitos), trombócitos e proteína plasmática total. Este estudo traz, pela primeira vez, resultados referentes a RDW e a contagem de trombócitos para a espécie, além de fornecer um tamanho amostral maior que estudos anteriores. Os intervalos, demonstrados neste estudo, relatam valores diferentes dos considerados normais para outras espécies de papagaios e, consequentemente, vêm contribuindo para embasar a conservação dessa espécie ameaçada de extinção, de grande valor biológico.(AU)

Detecção das encefalites virais zoonóticas com ênfase no vírus do Nilo Ocidental (WNV) em equídeos no Brasil

As doenças neurológicas nos equídeos constituem importantes causas de óbitos nestes animais. Entre as diversas etiologias das afecções neurológicas o vírus do Nilo Ocidental (do inglês West Nile virus-WNV), se destaca como importante agente zoonótico responsável por graves epidemias em todo mundo. Este arbovírus (arthropod borne viírus) está amplamente distribuído pelos continentes e mantém seu ciclo de transmissão através da relação entre mosquito vetor e aves reservatório/amplificadoras. Os equídeos, assim como o ser humano, são hospedeiros terminais não desenvolvendo viremia suficiente para transmissão ao vetor. Contudo, estes animais são considerados sentinelas, indicando a presença deste e de outros importantes arbovírus em uma região. A rápida expansão do WNV no continente americano, após sua entrada nos EUA em 1999, representou um desafio no diagnóstico deste vírus nos rebanhos equídeos. Evidências sorológicas indicam a presença do WNV no Brasil desde 2009. O país possui hoje um dos maiores rebanhos equídeos do mundo, apresentando condições ideais, além do clima, presença dos vetores e espécies de aves susceptíveis, para a epidemiologia de WNV e de outras importantes arboviroses. No presente estudo, foi padronizada uma técnica de nested RT-PCR, utilizando primers específicos direcionados à parte da região NS5 do WNV, com a finalidade de aumentar a sensibilidade do teste de diagnóstico molecular e, a partir dessa padronização, investigou-se a etiologia de quadros neurológicos em quatro equídeos (dois equinos e dois asininos) que vieram a óbito com encefalomielite aguda no Estado do Espírito Santo. Obteve-se então a primeira identificação do RNA de WNV em território nacional (capítulo 2). Após, a partir da parceria com o órgão de defesa animal do Estado de Minas Gerais, Laboratório de Sanidade Animal (Instituto Mineiro de Agropecuária, LSA-IMA), foram realizados novos estudos em 76 amostras de sistema nervoso central de equídeos com síndrome neurológica, todas negativas para o vírus da raiva, objetivando o diagnóstico das principais encefalites equinas virais através de técnicas moleculares. O diagnóstico diferencial incluiu o Herpesvírus equino tipo 1 (EHV-1- equine herpesvirus type 1); vírus do gênero Flavivirus, como WNV e Encefalite de Saint Louis (SLEV- saint louis encephalitis virus) e os vírus do gênero Alphavírus, como o vírus da encefalite equina do leste (EEEV- eastern equine encephalitis virus), o vírus da encefalite equina do oeste (WEEV- western equine encephalitis virus) e o vírus da encefalite equina venezuelana (VEEV-venezuelan equine encephalitis virus). As análises incluíram também histopatologia do SNC e de outros órgãos em 16 amostras. Os dados dos Formulários de Solicitação de Exame de Síndrome Neurológica enviados também foram avaliados quanto a parâmetros relacionados aos animais e as amostras. Como resultado, todas as amostras foram negativas para os flavivírus e alfavírus testados e 5% das amostras foram positivas para EHV-1. Contudo, foram constatadas diversas questões quanto à coleta, conservação e entrega ao laboratório, além da ausência de informações fundamentais, com apenas 43% dos formulários devidamente preenchidos (capítulo 3). Assim, com o objetivo de orientar e auxiliar os médicos veterinários quanto à adequada coleta e conservação de amostras de equídeos com síndrome neurológica, bem como na anamnese e exame clínico, orientações foram elaboradas para contribuir no diagnóstico das doenças neurológicas em equídeos (capítulo 5). Além disso, como estratégia alternativa no diagnóstico da infecção pelo WNV em equídeos, a investigação do vírus nas hemácias de um cavalo com síndrome neurológica resultou na detecção de um animal positivo na região sudeste do país (capítulo 4). Os resultados deste trabalho poderão auxiliar nas estratégias de diagnóstico das encefalites virais nos equídeos e na vigilância epidemiológica de WNV no Brasil, podendo ser valiosas no contexto da Saúde Única.

Neurological diseases in equidae are important causes of death in these animals. Among the various etiologies of neurological disorders, the West Nile virus-WNV, stands out as an important zoonotic agent responsible for serious epidemics worldwide. This arbovirus (arthropod borne virus) is widely distributed across continents and maintains its transmission cycle through the relationship between vector mosquito and reservoir / amplifying birds. Equidae, like humans, are terminal hosts not developing enough viremia to transmit to the vector. However, these animals are considered sentinels, indicating the presence of this and other important arboviruses in a region. The rapid expansion of WNV in the American continent, after its entry into the USA in 1999, represented a technical challenge in the diagnosis of infection in equine herds. In Brazil, one of the largest equine herds in the world, serological evidence indicated the presence of WNV since 2009. The country has ideal climate conditions for WNV transmission, as well as other arboviruses, with presence of vectors and the potential avian reservoirs species. In the present study, a nested RT-PCR technique was standardized, using specific primers directed to the NS5 gene of the WNV. The new technique was standardized investigating four equids (two horses and two donkeys) who died with neurologic signs of acute encephalomyelitis in the state of Espírito Santo. The study resulted in the first identification of WNV RNA in Brazil (chapter 2). Afterwards, based on the partnership with the animal defense agency of the State of Minas Gerais, through the Animal Health Laboratory (Instituto Mineiro de Agropecuária, LSA-IMA), further studies were carried out with 76 CNS samples from equines with neurological syndrome, all shown negative for rabies, aiming at the diagnosis of the main viral equine encephalitis through molecular techniques. The differential diagnosis included equine herpesvirus type 1 (EHV-1); viruses of the genus Flavivirus, such as WNV and Saint Louis Encephalitis (SLEV) and viruses of the Alphavirus genus, such as the Eastern Equine Encephalitis Virus (EEEV), the West Equine Encephalitis Virus (WEEV) and the Equine Encephalitis Virus Venezuelan (VEEV). The analyzes also included histopathology of the CNS and other organs in 16 samples. The Neurological Syndrome Examination Request Forms data sent were also evaluated for parameters related to animals and samples. As a result, all samples were negative for the flaviviruses and alphaviruses tested and 5% of the samples were positive for EHV-1. However, several issues were noted concerning the collection, conservation and delivery to the laboratory, and the absence of key information, with only 43% of forms properly filled (chapter 3). Thus, with the objective of guiding and assisting veterinarians in the field regarding the adequate collection and conservation of samples from equines with neurological syndrome, as well as aspects such as anamnesis and clinical examination, guidelines were developed to contribute to the diagnosis of neurological diseases in equines (chapter 5). In addition, as an alternative strategy in the diagnosis of WNV infection in equidae, the investigation of the virus in the red blood cells of a horse with neurological syndrome resulted in the detection of a positive animal in the southeastern region of the country (chapter 4). The results of this work may assist in the diagnostic strategies for viral encephalitis in equines and in the epidemiological surveillance of WNV in Brazil, which can be valuable in the context of One Health

Assessment of a probiotic containing bacillus subtilis on the performance and gut health of laying japanese quails (Coturnix Coturnix Japonica)

The present study was carried out to determine the effects of the inclusion of a spore-forming probiotic (Bacillus subtilis) in laying Japanese quail diets as an alternative to growth-promoting antibiotics to help produce healthy eggs and meat. This experiment was conducted as a completely randomized design with three treatments (control, 0.05% bacitracin methylene disalicylate (BMD), or 0.1% Bacillus subtilis) of five replicates of 11 quails each. Feed intake and egg production were recorded daily on cage basis. Body weight was determined at the beginning and end of the trial (36 and 42 weeks). At the end of the experiment (42 weeks), antibodies against Newcastle disease and avian influenza, egg components, Haugh units, eggshell quality and breaking strength, blood parameters, cecal microbial population, villus length, and crypt depth were measured. The dietary inclusion of Bacillus subtilis and BMD significantly (p0.05) increased egg production and egg weight; however, eggshell thickness and breaking strength, Haugh units, and eggshell percentages were not affected. The dietary addition of both products significantly (p0.05) decreased plasma cholesterol levels and increased LDL levels, as well as antibody levels against Newcastle disease and avian influenza (p0.01). In birds fed Bacillus subtilis and BMD, crypt depth was reduced, but villus height and villus to crypt ratio were significantly increased (p0.001) compared with those fed the basal diet. Cecal coliforms, E. coli, and Salmonella counts were reduced (p0.01) in quails fed the diets containing Bacillus subtilis and BMD compared those quails fed the non-supplemented diet. The results of this study demonstrated that in absence of AGPs, the inclusion of a spore-forming probiotic partially improves the performance of laying quails.

Assessment of a probiotic containing bacillus subtilis on the performance and gut health of laying japanese quails (Coturnix Coturnix Japonica)

The present study was carried out to determine the effects of the inclusion of a spore-forming probiotic (Bacillus subtilis) in laying Japanese quail diets as an alternative to growth-promoting antibiotics to help produce healthy eggs and meat. This experiment was conducted as a completely randomized design with three treatments (control, 0.05% bacitracin methylene disalicylate (BMD), or 0.1% Bacillus subtilis) of five replicates of 11 quails each. Feed intake and egg production were recorded daily on cage basis. Body weight was determined at the beginning and end of the trial (36 and 42 weeks). At the end of the experiment (42 weeks), antibodies against Newcastle disease and avian influenza, egg components, Haugh units, eggshell quality and breaking strength, blood parameters, cecal microbial population, villus length, and crypt depth were measured. The dietary inclusion of Bacillus subtilis and BMD significantly (p0.05) increased egg production and egg weight; however, eggshell thickness and breaking strength, Haugh units, and eggshell percentages were not affected. The dietary addition of both products significantly (p0.05) decreased plasma cholesterol levels and increased LDL levels, as well as antibody levels against Newcastle disease and avian influenza (p0.01). In birds fed Bacillus subtilis and BMD, crypt depth was reduced, but villus height and villus to crypt ratio were significantly increased (p0.001) compared with those fed the basal diet. Cecal coliforms, E. coli, and Salmonella counts were reduced (p0.01) in quails fed the diets containing Bacillus subtilis and BMD compared those quails fed the non-supplemented diet. The results of this study demonstrated that in absence of AGPs, the inclusion of a spore-forming probiotic partially improves the performance of laying quails. (AU)

Hematological and parasitological health conditions of the Pale-breasted Thrush (Turdus leucomelas) (Passeriformes: Turdidae) in southeastern Brazil

From an ecological point of view, parasites are considered important selective forces that influence all aspects of a host's life. The purpose of this study was to assess the health state of Turdus leucomelas Vieillot, 1818 (Turdidae: Passeriformes) inhabiting a Brazilian Cerrado in the breeding and molting seasons through a hematological parameter analysis and an evaluation of infection by blood parasites. The birds were collected with mist-nets, ringed and blood sampled to assess hematological parameters (hematocrit, hemoglobin concentration and white blood cells) and blood parasites. We detected blood parasites through optical microscopy and subsequently used PCR (amplification of the 18sRNA gene) to verify the presence of Plasmodium spp. (avian malaria). During the breeding season, the hemoglobin level and CHGM percentage were greater. Global leukocyte count was positively related to hemoglobin level, CHGM percentage and body weight. Of the total 31 T. leucomelas individuals examined, 18 presented Plasmodium parasites (58% of prevalence). There was a significant relationship between the presence of Plasmodium spp. and decreased CHGM. These results suggest a connection between the health parameters of wild birds and the physiological stress associated with the breeding season.

Hematological and parasitological health conditions of the Pale-breasted Thrush (Turdus leucomelas) (Passeriformes: Turdidae) in southeastern Brazil

From an ecological point of view, parasites are considered important selective forces that influence all aspects of a host's life. The purpose of this study was to assess the health state of Turdus leucomelas Vieillot, 1818 (Turdidae: Passeriformes) inhabiting a Brazilian Cerrado in the breeding and molting seasons through a hematological parameter analysis and an evaluation of infection by blood parasites. The birds were collected with mist-nets, ringed and blood sampled to assess hematological parameters (hematocrit, hemoglobin concentration and white blood cells) and blood parasites. We detected blood parasites through optical microscopy and subsequently used PCR (amplification of the 18sRNA gene) to verify the presence of Plasmodium spp. (avian malaria). During the breeding season, the hemoglobin level and CHGM percentage were greater. Global leukocyte count was positively related to hemoglobin level, CHGM percentage and body weight. Of the total 31 T. leucomelas individuals examined, 18 presented Plasmodium parasites (58% of prevalence). There was a significant relationship between the presence of Plasmodium spp. and decreased CHGM. These results suggest a connection between the health parameters of wild birds and the physiological stress associated with the breeding season.

Hematological and parasitological health conditions of the Pale-breasted Thrush (Turdus leucomelas) (Passeriformes: Turdidae) in southeastern Brazil

From an ecological point of view, parasites are considered important selective forces that influence all aspects of a host's life. The purpose of this study was to assess the health state of Turdus leucomelas Vieillot, 1818 (Turdidae: Passeriformes) inhabiting a Brazilian Cerrado in the breeding and molting seasons through a hematological parameter analysis and an evaluation of infection by blood parasites. The birds were collected with mist-nets, ringed and blood sampled to assess hematological parameters (hematocrit, hemoglobin concentration and white blood cells) and blood parasites. We detected blood parasites through optical microscopy and subsequently used PCR (amplification of the 18sRNA gene) to verify the presence of Plasmodium spp. (avian malaria). During the breeding season, the hemoglobin level and CHGM percentage were greater. Global leukocyte count was positively related to hemoglobin level, CHGM percentage and body weight. Of the total 31 T. leucomelas individuals examined, 18 presented Plasmodium parasites (58% of prevalence). There was a significant relationship between the presence of Plasmodium spp. and decreased CHGM. These results suggest a connection between the health parameters of wild birds and the physiological stress associated with the breeding season.

PERFIL SANITÁRIO DE FILHOTES DE Amazona brasiliensis DE VIDA LIVRE NO PARANÁ: PARÂMETROS HEMATOLÓGICOS, BIOQUÍMICOS E PESQUISA DE AGENTES INFECCIOSOS

Atualmente, há maior necessidade de pesquisas de doenças e estabelecimento de perfis hematológicos e bioquímicos em populações de aves ameaçadas de extinção. O acompanhamento da saúde desses animais deve ser encorajado pela crescente evidência de que certas enfermidades podem causar impactos adversos nessas populações, e estudos são escassos. O papagaio-de-cara-roxa (Amazona brasiliensis) é considerado uma espécie vulnerável, habitando a floresta atlântica do litoral sul de São Paulo, litoral do Paraná e litoral norte de Santa Catarina. Desde 1997, a Sociedade de Pesquisa em Vida Selvagem e Educação Ambiental desenvolve o Projeto de Conservação do Papagaio-de-cara-roxa, numa tentativa de minimizar o declínio da população. Informações sobre valores hematológicos, bioquímicos e pesquisa de patógenos nesta espécie são raros. O presente trabalho foi dividido em três capítulos. O capítulo 1 objetivou estabelecer intervalos de referência para parâmetros hematológicos e bioquímicos em 37 filhotes saudáveis de A. brasiliensis de vida livre no estado do Paraná, sul do Brasil e comparar os resultados entre sexos. O capítulo 2 objetivou determinar os mesmos parâmetros em filhotes saudáveis e doentes e comparar os resultados entre grupos que apresentaram diferentes sinais clínicos com um grupo saudável clinicamente. O capítulo 3 objetivou determinar a microbiota de 38 filhotes, comparando-a entre aves provenientes de dois tipos de ninhos (madeira e PVC) e pesquisar patógenos por reação em cadeira da polimerase (Chlamydophila psittaci, vírus da influenza aviária, da doença de Newscastle e da Febre do oeste do Nilo) e anticorpos para C. psittaci pelo teste de ELISA modificado, em 50 e 64 indivíduos, respectivamente. As coletas de sangue foram realizadas por meio de seringas e agulhas de insulina estéreis, e amostras de cloaca e orofaringe foram obtidas por swabs estéreis. Médias significativamente superiores para colesterol foram encontradas em fêmeas. O exame físico indicou 37 aves clinicamente sadias, 26 com anormalidades discretas e 5 com alterações graves, e os papagaios foram divididos em 7 grupos de acordo com os achados clínicos. Médias significativamente diferentes entre os grupos foram observadas para os parâmetros hemoglobina, volume globular médio, heterófilo absoluto, relação heterófilo:linfócito, proteína plasmática total, proteína total, glicose, lactato desidrogenase, creatina quinase, globulinas, cálcio e fósforo. Foram isoladas 125 colônias bacterianas com identificação de 19 espécies e três gêneros. As espécies com maior prevalência de isolamento foram Escherichia coli (70,73%), Klebsiella oxytoca (39,13%), Klebsiella pneumoniae (30,43%) e Enterobacter aerogenes (30,43%). Samonella enteritidis foi detectada em três amostras. A microbiota dos papagaios não diferiu em relação ao tipo de ninho. Todas as amostras para pesquisa de patógenos e anticorpos foram negativas. Os resultados possibilitaram a obtenção de dados ainda não descritos para a espécie no estado do Paraná e o fornecimento de um ponto de referência para futuras avaliações e interpretações de resultados laboratoriais, contribuindo com planos de conservação.

There is a greater need for an increase effort in research on disease and determination of hematologic and biochemical profiles in endangered avian populations. Health monitoring of these animals should be encouraged in light of increasing evidence that disease has an adverse impact on avian populations. The red-tailed Amazon parrot (Amazona brasiliensis) is a vulnerable species and is endemic in the Atlantic Forest, inhabiting coastal regions in the south-east Brazilian states of São Paulo, Parana and Santa Catarina. A conservation project for A. brasiliensis has been developed by the non-profitable organization Society for Wildlife Research and Environmental Education since 1997, to minimize the population decline. Information about hematology, biochemical and diseases in this species are rare. This study was split into three chapters. Chapter 1 aimed to determine reference intervals for hematologic and biochemical parameters of 37 free-living A. brasiliensis healthy nestlings in Parana state, southern Brazil, and compare the results between sexes. Chapter 2 aimed to determine the same parameters in healthy and sick nestlings and compare the results between groups with different clinical findings. Chapter 3 aimed to determine the microbiota of 38 nestlings, comparing the results between birds from two types of nests (wood and PVC); also aimed to detect pathogens by polymerase chain reaction (Chlamydophila psittaci, avian influenza virus, Newcastle disease virus and west Nile virus) and to detect antibodies to C. psittaci by modified ELISA test, in 50 and 64 parrots, respectively. Blood samples were collected in sterile syringes using insulin needles, and cloacal and oropharyngeal samples were collected using sterile swabs. Significantly higher values of cholesterol were observed in females. Physical examinations indicated that 37 birds were clinically healthy, 26 had mild abnormalities and 5 had severe disorders, and the parrots were split into 7 groups according to clinical findings. Significantly different means were observed for hemoglobin, mean corpuscular hemoglobin concentration, absolute heterophil, heterophil:lymphocyte ratio, total plasma protein, total protein, glucose, lactate dehydrogenase, creatine kinase, globulin, calcium and phosphorus. 125 bacterial colonies were isolated and 19 species and 3 genera were identified. The species with the highest prevalence were Escherichia coli (70.73%), Klebsiella oxytoca (39.13%), Klebsiella pneumoniae (30.43%) and Enterobacter aerogenes (30.43%). Salmonella enteritidis was detected in three samples. The microbiota of parrots did not differ between the types of nest. All samples tested by ELISA and PCR were negative. The results allowed obtaining data not yet described for this species in Parana state, providing a reference point for future assessments and interpretations of laboratory results, contributing to the conservation plans.

Epidemiologia molecular, filogenia e filogeografia do Infectious bronchitis virus em um panorama global

A criação de aves em sistemas industrializados vem adquirindo, mundialmente, características que envolvem uma elevada densidade animal, o que, consequentemente, leva ao surgimento de condições epidemiológicas capazes de predisporem ao aparecimento de problemas sanitários. Diante deste contexto, aumenta-se a preocupação com as doenças infecciosas, dentre elas, a Bronquite Infecciosa das Galinhas (BIG), cujo agente etiológico é o Infectious bronchitis virus (IBV). O presente estudo teve como objetivo reconstruir a epidemiologia molecular do IBV, por meio de análises filogenéticas e filogeográficas dos genes estruturais N e S1, para estudo da evolução viral. Estas informações são importantes para estabelecer um padrão de dispersão em um contexto histórico geográfico, entender eventos biológicos no passado relacionados à dinâmica populacional e ajudar no desenvolvimento de estratégias efetivas de controle do IBV. Para isso, foram montados dois bancos de dados com sequências moleculares completas e parciais dos genes N e S1 obtidos no GenBank. O primeiro banco de dados (IBV1) contemplou sequências completas de S1 (N=256) e N (N=154) e o segundo banco de dados (IBV2) foi montado com sequências parciais de S1 (N=390) e N (N=173). Importantes parâmetros para estudo da evolução viral foram analisados nesse trabalho para os genes S1 e N do IBV, incluindo estimativas de eventos de recombinação, taxa de mutação, pressão de seleção, dinâmica populacional e dispersão espacial. Os resultados apontam que, na ausência de uma elevada taxa de mutação e pelo fato da seleção purificadora ser a principal forma de seleção natural no IBV, o processo de recombinação tem um papel fundamental na evolução do IBV. As análises de dinâmica populacional sugerem que, até meados da década de 2000, os diferentes programas de vacinação, juntamente com os métodos de biosseguridade, foram eficazes para controlar o crescimento do tamanho efetivo populacional do IBV. Contudo, a partir de 2007, o aumento da população de variantes do IBV, pode indicar que os protocolos vacinais, usados no presente, não estão sendo mais eficazes. Os resultados das análises de dispersão viral do gene S1 e N ixforam analisados de forma conjunta, a fim de reconstruir um padrão de dispersão do IBV ao longo de sua história evolutiva. O ponto de origem de todas as cepas atuais do IBV foi a China que, após dispersão local, disseminou para os Estados Unidos e, por conseguinte, se dispersou para diferentes países e continentes até alcançar a ampla distribuição mundial encontrada nos dias atuais. Os principais centros de dispersão atuais do IBV foram a China, Estados Unidos, Brasil, Taiwan e Europa.

The intensification of poultry in industrial systems has acquired, worldwide, features that involve a high stock density, which, consequently, leads to the emergence of epidemiological conditions that may predispose to the appearance of health problems. Given this context, the concern with infectious diseases increases, among the diseases is the Avian Infectious Bronchitis (IB), whose etiologic agent is the Infectious bronchitis virus (IBV). This study aimed to reconstruct the molecular epidemiology of IBV through phylogenetic and phylogeographic analyzes of structural genes N and S1, to the study of viral evolution. These pieces of information are important to establish a pattern of dispersion in a geographical historical context, to understand biological events in the past related to population dynamics and to help develop effective strategies for control of IBV. To establish a better outlook for discussion of IBV molecular epidemiology, there were created two databases with complete and partial molecular sequences of genes N and S1 obtained from GenBank. The first database (IBV1) considered complete sequences of the S1 (N = 256) subunit and of the N (N = 154) gene of IBV. To increase the extension of countries from which sequences are available, a second database (IBV2) was created with partial sequences of the S1 (N = 390) gene and the N (N = 173) gene. Important parameters for the study of viral evolution were analyzed in this work for the S1 and N genes of the IBV, including estimates of recombination events, mutation rate, selection pressure, population dynamics and spatial dispersion. The results show that, in the absence of a high mutation rate and because of the purifying selection to be the main form of natural selection in IBV, the recombination process plays a key role in the evolution of IBV. The analysis of population dynamics suggest that, until the mid-2000s, the different vaccination programs, along with the biosecurity methods have been effective in controlling the growth of effective population size of the IBV. However, since 2007, the increase in the population of IBV variants may indicate that the vaccine protocols, used in the present, have not been effective anymore. The results of analyzes xiof the viral spread of the gene S1 and N were analyzed together, in order to reconstruct a pattern of the IBV dispersion along its evolutionary history. The point of origin of all current strains of IBV was China, which after local dispersion spread to the United States of America and thus dispersed to different countries and continents until reaching the worldwide distribution found today. The current main scattering centers of the IBV were China, the United States, Brazil, Taiwan and Europe.

Hematologia, pesquisa de hemoparasitos e mensuração da atividade de colinesterases plasmáticas em Falconiformes e Strigiformes do Estado de São Paulo, Brasil / Hematology, survey for hemoparasites and measurement of plasmatic cholinesterases activity of Falconiformes and Strigiformes of São Paulo State, Brazil

Foram coletadas 137 amostras sanguíneas de 132 espécimes diferentes, pertencentes a sete espécies de Strigiformes (n=92) e onze espécies de Falconiformes (n=45), entre os meses de setembro de 2008 e julho de 2009. As amostras foram submetidas a hemograma completo, pesquisa de hemoparasitos através da avaliação do esfregaço sanguíneo, sexagem através da técnica de PCR e mensuração da atividade de colinesterases plasmáticas através do método de Ellman (1961). Apenas sete corujinhas-do-mato (Megascops choliba) apresentaram hemoparasitos (5,3 % do total de rapinantes). Houve diferença significativa conforme o estado de saúde da ave, sendo que a todos os animais clinicamente hígidos não estavam infectados, porém 20,4% dos animais com desordens clínicas apresentavam hemosporídeos. Quantitativamente, a parasitemia mediana dos animais infectados foi de 3,8 hemácias parasitadas por 10.000 hemácias, um índice de infecção relativamente baixo. A grande maioria dos parâmetros hematológicos não apresentou diferenças estatísticas em função da presença ou ausência de hemoparasitos, o que na maioria dos casos caracterizou uma infecção branda e pouco patogênica. A atividade de colinesterases plasmáticas apresentou uma distribuição normal, com uma média geométrica de 0,8333 micromoles/mL e desvio padrão geométrico de 1,7087 µmols/mL. A atividade de colinesterases plasmáticas apresentou diferenças estatísticas significativas entre Falconiformes (0,5705±1,6497 µmols/mL) e Strigiformes (1,0070±1,5620 µmols/mL). Não houve diferenças significativas entre machos ou fêmeas e adultos ou jovens dentre os Falconiformes ou Strigiformes. Falconiformes não hígidos apresentaram valores surpreendentemente mais baixos do que os clinicamente hígidos. Entre os Strigiformes, os espécimes de cativeiro apresentaram valores mais baixos de atividade de colinesterases plasmáticas do que os de vida livre. A atividade de colinesterases plasmáticas foi significativamente diferente entre os animais com hemoparasitos (mediana geométrica = 2,0132 µmols/mL) e sem hemoparasitos (mediana geométrica = 0,8331 µmols/mL). Este estudo contribui com informações relevantes sobre a ocorrência de hemosporídeos em aves de rapina do Estado de São Paulo, alertando as Instituições sobre a necessidade de monitoração destes protozoários sanguíneos. Além disso, contribui com dados inéditos sobre a atividade de colinesterases plasmáticas em rapinantes

137 blood samples were collected from 132 different specimen, belonging to seven species of Strigiformes (n=92) and eleven species of Falconiformes (n=45), from September 2008 to July 2009. Samples were submitted to complete hemogram, survey for hemoparasites through evaluation of blood smears, sex identification by PCR and measurement of plasmatic cholinesterases by the method of Ellman (1961). Only seven black-capped-screech-owls (Megascops choliba) were positive for hemoparasites (5,3% of total raptors). There were significative difference according to avian health status; all the clinicaly higid animal were not infected, but 20,4% of animals with clinical desorders presented haemosporidians. Quantitatively, the average parasitemia was 3,8 parasited erythrocytes in 10.000 erythrocytes, a infection rate relatively low. The most of hematologic parameters did not present statistic differences in function of presence or absence of hemoparasites, what in most of the cases characterized a low and not much pathogenic infection. The plasmatic cholinesterase activity presented normal distribution, with geometric average of 0,8333 micromoles/mL and geometric standard deviation of 1,7087 µmols/mL. The plasmatic cholinesterase activity presented statistic differences between Falconiformes (0,5705±1,6497 µmols/mL) and Strigiformes (1,0070±1,5620 µmols/mL). There were not significative differences between males of females and adults or youngs between Falconiformes or Strigiformes. Non-higid Falconiformes presented surprisiling much low values than those clinically higid. Between Strigiformes, the captivity specimens present much lower plasmatic cholinesterase activity values than free ranging animals. The plasmatic cholinesterase activity was signifactively different among animal with hemoparasites (geometric average = 2,0132 µmols/mL) and without hemoparasites (geometric average = 0,8331 µmols/mL). This research contributes with relevant information about the haemosporidian occurence in raptors of State of São Paulo, alerting the Institutions about the necessity of monitorizing haemoprotozoa. Besides that, contributes with unpublished data about the plasmatic cholinesterase activity in raptors

Effects of mannan-oligosaccharide supplementation on gut health, immunity, and production performance of broilers / Efeitos da suplementação de mananoligossacarídeo na saúde do intestino, imunidade e desempenho da produção de frangos

The study was designed to investigate the effect of mannan-oligosaccharide (MOS) supplementation on intestinal histomorphology, immunity against Newcastle disease virus (NDV) and productive parameters of broilers. A total of 1800, day old broiler chicks of Cobb-500 strain were selected and randomly assorted into 6 treatment groups: T1 (basal diet without antibiotics as negative control); T2 (basal diet plus antibiotics as positive control group); T3 (basal diet plus 200g/ton MOS); T4 (basal diet plus 400g/ton MOS); T5 (basal diet plus 600g/ton MOS) and T6 (basal diet plus 800g/ton MOS). Each treatment was having 6 replicates and the feed intake, body weight gain and feed conversion ratio (FCR) were recorded on weekly basis. Results showed that, MOS supplemented birds have significantly higher feed intake, weight gain and FCR (P < 0.05). Similarly, supplementation of MOS showed positive effect on villus height and crypt depth both in jejunum and ilium. Goblet cell density was unaffected by MOS addition (P < 0.05). Furthermore, birds fed with diets containing MOS, exhibited better productive performance in comparison to positive and negative control groups. In conclusion, MOS can replace antibiotic growth promoters (AGPs) as non-microbial performance-enhancing feed advocates.

O estudo foi desenhado para investigar o efeito da suplementação de mananoligossacarídeo (MOS) na histomorfologia intestinal, imunidade contra o vírus da doença de Newcastle (NDV) e parâmetros produtivos de frangos de corte. Um total de 1.800 pintos de corte de um dia da linhagem Cobb-500 foram selecionados e distribuídos aleatoriamente em 6 grupos de tratamento: T1 (dieta basal sem antibióticos como controle negativo); T2 (dieta basal mais antibióticos como grupo controle positivo); T3 (dieta basal mais 200g/ton MOS); T4 (dieta basal mais 400g/ton MOS); T5 (dieta basal mais 600g/ton MOS) e T6 (dieta basal mais 800g/ton MOS). Cada tratamento tinha 6 repetições e o consumo de ração, ganho de peso corporal e conversão alimentar foram registrados semanalmente. Os resultados mostraram que as aves suplementadas com MOS apresentam consumo de ração, ganho de peso e CA significativamente maiores (P < 0,05). Da mesma forma, a suplementação de MOS mostrou efeito positivo na altura das vilosidades e na profundidade das criptas tanto no jejuno quanto no íleo. A densidade de células caliciformes não foi afetada pela adição de MOS (P < 0,05). Além disso, as aves alimentadas com dietas contendo MOS apresentaram melhor desempenho produtivo em comparação aos grupos controle positivo e negativo. Em conclusão, o MOS pode substituir os promotores de crescimento de antibióticos (AGPs) como defensores de alimentos não microbianos que melhoram o desempenho.