Este estudo teve como objetivo pesquisar a ocorrência e diversidade molecular de coronavírus aviários em codornas e galinhas criadas nas mesmas propriedades e determinar o papel epidemiológico das codornas na bronquite infecciosa das galinhas (BIG). Para isso, foram coletados em granjas localizadas no estado de São Paulo e Espírito Santo pools de aparelho reprodutivo, pulmões, rins, traquéia e conteúdo entérico de codornas e galinhas com histórico de manifestações clínicas compatíveis com BIG. Estas amostras foram testadas para coronavírus aviário mediante uma semi-nested RT-PCR dirigida a região não-traduzida 3 (3´UTR) e as amostras positivas foram submetidas a RT-PCR do gene codificador da proteína RNA-polimerase RNA-dependente (RdRp) e duas RT-PCR, incluindo uma tipo multiplex dirigidas a proteína de espícula S do vírus da BIG, para genotipagem. Amplicons ou fragmentos amplificados da 3\'UTR (a partir de amostras de codorna) foram clonados e sequenciados. Outras duas RT-PCR foram utilizadas para detecção de metapneumovírus aviário (aMPV) e o vírus da doença de Newcastle (NDV). Coronavírus aviários foram encontrados em todos os tipos de amostras estudadas em galinhas e codornas criadas nas mesmas propriedades, sendo que aMPV subtipo B foi encontrado em galinhas e o NDV não foi encontrado em nenhuma amostras. Todos os coronavírus aviários encontrados, foram classificados como variantes pela multiplex RT-PCR, não sendo entretanto, obtidas sequências de DNA para o gene S. Com base em sequências de DNA para os genes codificadores da proteína RdRp e da região 3´UTR pode-se demonstrar que as codornas estudadas apresentaram o coronavírus aviário identificados como próximo àqueles relacionados à bronquite infecciosa das galinhas, havendo diversidade molecular filogeográfica para os vírus de codornas. Desta forma, sugere-se que as codornas podem servir como reservatórios para coronavírus aviários onde haja criações em proximidade com outras espécies aviárias. / This study aimed to investigate the occurrence and molecular diversity of avian coronavirus in quails and laying hens, raised on the same farms and determine the role of quails in the epidemiology of avian infectious bronchitis (IB). To this end, pools of lungs, trachea, female reproductive tracts, kidneys and enteric contents were collected from quails and laying hens flocks with IB-like symptoms, co-housed in farms located in Sao Paulo and Espírito Santo states, Brazil, during 2009-2010. Chickens and quails samples were screened for IBV with an RT-PCR to the 3UTR and positive samples were submitted to RT-PCRs to the RNA-dependent RNA-polymerase gene (RdRp) and two different RT-PCRs to the spike gene, including a typing-multiplex one. Amplicons of 3UTR (from quails samples) were cloned and sequenced. Two other RT-PCRs were used to detect the avian metapneumovirus (aMPV) and Newcastle disease virus (NDV). Avian coronavirus was found in all types of samples analyzed in chickens and quails raised on the same farms, aMPV subtype B was found in chickens and the NDV was not observed in any samples. All avian coronavirus found were classified as variants by multiplex RT-PCR, however, DNA sequences for gene S were not obtained. Based on the DNA sequences for genes encoding the protein RdRp and the 3\'UTR region can be show that avian coronavirus in quails are closely related to avian infectious bronchitis virus, with a molecular phylogeographic diversity for quails viruses; thus, quails might act as reservoirs for avian coronaviroses when in close contact with other avian species.

A partir de amostras de campo de diferentes regiões do país, o TCoV foi identificado em aves de 13-84 dias, em órgãos como intestino, rins e traqueia, por uma reação em cadeia pela polimerase para a região 3\' UTR e o gene 3. A presença deste coronavírus nesses diferentes órgãos, que não o intestino, sugere a presença de uma diferente estirpe, com características fenotípicas mais próximas em relação ao vírus da Bronquite Infecciosa das Galinhas (BIG), o IBV. Ao mesmo tempo, um perfil considerando a idade das aves positivas foi desenhado por um teste T de duas amostras, que mostrou maior positividade em aves mais jovens. Com a finalidade de desenvolver um novo ensaio de PCR, várias sequências do TCoV disponíveis no GenBank foram alinhadas, um segmento gênico foi desenhado e submetido a diferentes condições de reação. Um fragmento de 436 pb foi obtido, sequenciado e apresentou 100% de identidade quando sua sequência foi comparada a sequência completa do TCoV. A reação mostrou boa repetibilidade, bem como boa reprodutibilidade, e algumas amostras de campo não se apresentaram positivas. / TCOV was identified from field samples of birds with 13-84 days, from different regions of the country, and from organs such as intestine, kidneys and trachea, by polymerase chain reaction for 3\' UTR region and gene 3. The presence of this coronavirus in these different organs, not intestine, suggest a different strain, with similar phenotype observed in Infectious Bronchitis Virus (IBV). At the same time, a profile considering age of positive birds was designed by a Two sample T-test, which showed higher positivity in younger poults. Several TCOV sequences obtained from GenBank were aligned, and a genic segment was designed and submitted to different conditions in reaction in pursuit to develop a new PCR assay. A 436bp fragment was obtained, sequenced and presented 100% ID when compared to a TCOV complete sequence. The reaction featured good repetition, and reproduction as well, and some field samples did not turn out on positive results.

O setor avícola apresenta o maior crescimento em volume produzido dentre todos os setores cárneos no Brasil. A grande participação dos produtos avícolas na alimentação humana somada ao risco do desenvolvimento de resistência bacteriana, levaram a União Européia (UE) a abolir utilização de antimicrobianos como aditivos e de forma profilática na ração de animais. A remoção dos aditivos associada ao sistema de criação intensivo acabaram por fazer que doenças, até então consideradas controladas, se tornassem reemergentes. A enterite necrótica aviária (NE) é considerada um exemplo. De forma geral, condições estressoras predispõem ao desenvolvimento de doenças, sendo o calor um dos estressores mais comuns que ocorrem em granjas aviárias. Este estudo enfoca o efeito do estresse térmico por calor (35±1ºC) sobre o desenvolvimento da NE em frangos de corte. Para isso, 60 frangos de corte machos foram divididos em 6 grupos experimentais: 1 Grupo Controle; 2 Grupo Controle Estressado (C/HS35); 3 Grupo Tioglicolato (T); 4 Grupo Tioglicolato Estressado (T/HS35); 5 Grupo Infectado (I); 6 Grupo Infectado Estressado (I/HS35). A infecção experimental com Clostridium perfringens foi feita por via oral, com a bacteria em meio de cultura misturada a ração, do 15º ao 21º dia de vida nos grupos I e I/HS35. O estresse por calor (35±1º C) foi realizado do 14º ao 21º dia de vida das aves dos grupos estressados. Durante todo período experimental os animais foram mantidos em isoladores. Em relação aos animais não estressados, os animais submetidos ao estresse por calor apresentaram: 1- menor escore lesional macro e microscópico no intestino delgado; 2- maior concentração de IgA no lavado intestinal do duodeno; 3 - menor concentração de IgA no jejuno; 4 - redução dos níveis séricos de IgA e IgY; 5 - maior concentração sérica de IgM; 6 - diminuição qualitativa evidente dos heterofilos intestinais em relação aos animais infectados e estressados. Portanto, mostrou-se que o estresse por calor apresentou efeito imunomodulador importante, ao reduzir a inflamação intestinal. Este achado associa-se provavelmente à diminuição da imunidade inata por redução da migração de heterófilos para a mucosa intestinal, desta forma prevenindo a manifestação de um um quadro clínico mais grave de NE, fato associado à diminuição das lesôes desencadeadas pelo processo inflamatório heterofílico. / The poultry sector presented the highest growth in the volume of production among all meat sectors in Brazil. The great participation of poultry products on human diet together with the risk of food and environmental contamination by resistant bacteria led the European Union (EU) countries to abolish the use of antibiotics as feed additives in animal production. This fact associated with the intensive farming system are being reported as responsible for the re-emergence of some already controlled diseases. The avian necrotic enteritis (NE) exemplify such an effect. Generally, stressful conditions are predisponent factors for disease development; heat stress is one of the most common stressor in poultry farms. This study focuses on the effects of heat stress (35 ± 1 º C) on the development of NE in broilers. For this purpose, 60 male broilers were divided into 6 groups: 1 - control group, 2 - stressed control group (C/HS35) 3 - thioglycolate group (T) 4 - thioglycolate stressed group (T/HS35); 5 - infected group (I) 6 - infected stressed group (I/HS35). Experimental infection with Clostridium perfringens, grown in thioglycollate broth medium, was given through the feed to the birds of groups I and I/HS35 from the 15th to 21st days of life. The heat stress (35 ± 1 °C) was induced continuously from the 14th to the 21st day of life in birds of the stressed groups. Throughout the experimental period the animals were kept in isolators. Compared to non-stressed animals, broilers subjected to heat stress showed: lower gross and microscopic score lesions in the small intestine; increased concentrations of IgA in duodenal lavage and decreased IgA concentrations in the jejunum; smaller concentrations of serum IgA and IgY; increased concentration of serum IgM; reduction in gut number of heterophils in the thioglycolate treated and in the infected groups. Therefore, this experimental model showed that heat stress presented a significant immunomodulatory role on the induced NE, most probably because it reduced intestinal inflammation via decrease in heterophils migration to the intestinal mucosa, which in turn might had reduced tissue damage during infamation, hence preventing the development of a more severe form of NE.

This study aimed to investigate the occurrence and molecular diversity of coronavirus in quail and laying hens raised on the same farms and quail only farms, to determine the role of quail as reservoir for avian infectious bronchitis virus (IBV). To this end, two investigations were carried out, one in the São Paulo state, Southeastern Brazil, in 2013, when some farmers started quail vaccination with Massachusetts IBV serotype after surveillance carried out in 2009-2010 and the other in two regions of Northern Italy, in 2015. In the Brazilian study, samples were collected as pools of tracheas, lungs, reproductive tract, kidneys and enteric contents from quail (Coturnix coturnix Japonica) and laying hens showing IB-like symptoms, while, in the Italian study, samples were collected as pools of tracheal and cloacal swabs and intestine/enteric content from European quail (Coturnix coturnix), showing enteric disorders. All samples were tested by a nested RT-PCR targeted to the 3\'UTR of the Gammacoronavirus genus. Positive samples were submitted to RT-PCR to the RNA-dependent RNA-polymerase gene (RdRp) and two different RT-PCRs to the spike gene, including a typing-multiplex one. Two other RT-PCRs were used to detect the avian metapneumovirus (aMPV) and Newcastle disease virus (NDV). Avian coronavirus was found in all types of samples analyzed in quail and chickens from both type of creations, aMPV subtype B was found in chickens (Brasil) and the NDV was not observed in any samples. Based on the DNA sequences for the RdRp gene, Brazilian and Italian quail strains clustered within either Gammacoronavirus or Deltacoronavirus genus, while, for one Brazilian sample, it was detected co-infection with the two genuses. Phylogeny based on partial S1 subunit sequences showed that the gammacoronaviruses detected in the Brazilian and Italian quail belong to the Brazil type and 793/B, respectively. These results suggest that quail are susceptible to Gamma and Deltacoronavirus and that quail avian coronavirus share spike genes identical to chicken infectious bronchitis virus (IBV); thus, quail might act as reservoirs for avian coronaviruses. Also, Massachusetts vaccination was not efficient to control IBV in Brazilian quail. / Este estudo teve como objetivo pesquisar a ocorrência e diversidade molecular de coronavírus em codornas e galinhas criadas nas mesmas propriedades e em codornas criadas em propriedades isoladas, para determinar o papel das codornas como reservatório para o vírus da bronquite infecciosa das galinhas (IBV). Para isso, duas pesquisas foram realizadas, uma em 2013, no estado de São Paulo, Sudeste do Brasil, onde algumas granjas iniciaram a vacinação em codornas contra o IBV com o sorotipo Massachusetts, após um estudo realizado em 2009-2010; e a outra, em 2015, em duas regiões do Norte da Itália. No estudo brasileiro, foram coletados pools de aparelho reprodutor, pulmões, rins, traqueia e conteúdo entérico de codornas (Coturnix coturnix japonica() e galinhas com histórico de manifestações clínicas compatíveis com a Bronquite Infecciosa das galinhas (BIG). Por outro lado, no estudo italiano, as amostras foram coletadas em forma de pools de swabs traqueais e cloacais e intestino/conteúdo entérico de codornas (Coturnix coturnix) com sinais entéricos. Estas amostras foram testadas para os coronavírus aviário (Gammacoronavirus) mediante uma semi-nested RT-PCR dirigida a região não-traduzida 3 (3´UTR). As amostras positivas foram submetidas a RT-PCR do gene codificador da proteína RNA-polimerase RNA-dependente (RdRp) e duas RT-PCRs, incluindo uma multiplex dirigidas a proteína de espícula (S) do vírus da BIG, para genotipagem. Além disso, a detecção de metapneumovírus aviário (aMPV) e o vírus da doença de Newcastle (NDV) também foi realizada por meio de RT-PCRs. Coronavírus aviários foram encontrados em todos os tipos de amostras estudadas em codornas e galinhas de todos os tipos de criações, aMPV subtipo B foi encontrado em galinhas (Brasil) e o NDV não foi encontrado em nenhuma amostras. Com base nas sequências de DNA para o gene codificador da proteína RdRp, as amostras brasileiras e italianas foram agrupadas no gênero Gamma ou Deltacoronavirus, enquanto que, em uma amostra brasileira, foi detectada coinfecção pelos dois gêneros. A filogenia com base nas sequências parciais da subunidade S1da proteína de espícula, evidenciou que os Gammacoronavirus detectados nas codornas brasileiras e italianas pertencem ao genótipo Brasil e 793/B, respectivamente. Estes resultados sugerem que as codornas são suscetíveis aos coronavírus do gênero Gamma e Delta e os coronavírus aviários das codornas compartilham genes de espícula idênticos aos do IBV. Desta forma, sugere-se que as codornas podem servir como reservatórios para coronavírus aviários e que a vacinação com o sorotipo Massachusetts não foi eficiente no controle de IBV nas codornas brasileiras.

Orientador: Helio Jose Montassier / Resumo: As infecções do trato respiratório de aves têm-se constituído em problemas crescentes e com marcantes consequências negativas sobre a produção avícola em várias partes do mundo, notadamente onde a avicultura é mais desenvolvida como no Brasil. Dentre essas enfermidades, destacam-se as micoplasmoses aviárias e a pneumovirose aviária, que, apesar de suas relevâncias em sanidade avícola, não têm sido investigadas de forma sistematizada no Brasil, em especial no que concerne à interação entre esses agentes ou a ocorrência de co-infecção em frangos de corte. O presente estudo investigou a ocorrência de infecção por Mycoplasma gallisepticum (MG) e pelos subtipos A ou B de Metapneumovírus aviário (AMPV) em frangos de corte de plantéis avícolas comerciais mantidos em granjas mais tecnificadas localizadas nas regiões Sudeste e Centro-Oeste do Brasil. Para tanto, as técnicas de PCR e de RT-Nested-PCR foram usadas na detecção e/ou identificação, respectivamente de MG e AMPV em amostras de suabes nasais e traqueais colhidos de 87 lotes de frangos de corte com problemas respiratórios e oriundos de 15 granjas de produção comercial de frangos de corte. Dos lotes amostrados, dois deles em um total de 87 (2,3%) e de uma única granja da região Sudeste, mostraram-se positivos para MG, enquanto que nenhum dos lotes investigados revelou-se positivo para AMPV. A baixa ou nenhuma incidência desses agentes pode ser explicada pela utilização de medidas cada vez mais efetivas para o controle sanitário... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Respiratory infections of poultry may be an increasing and negative problem with poultry production in several parts of the world, especially when poultry farming is more widely used than in Brazil. These diseases include avian mycoplasmosis and avian pneumovirosis, which, despite their relevance in poultry health, have not been systematically investigated in Brazil, especially with regard to the interaction between these agents or the occurrence of co-infection in broilers. The present study investigated the occurrence of infection by Mycoplasma gallisepticum (MG) and A or B subtypes of avian Metapneumovirus (AMPV) in broiler commercial poultry farms located in the Southeast and Center-West regions of Brazil. For this, PCR and RT-Nested-PCR techniques were used in the detection and / or identification, respectively of MG and AMPV in nasal and tracheal swab samples collected from 87 lots of broilers with respiratory problems and from 15 commercial production of broilers. Of the sampled lots, two of them in 87 (2.3%) were positive for MG, whereas none of the lots were positive for AMPV. The low or no incidence of these agents can be explained by the use of increasingly effective measures for the sanitary control of these agents in the commercial farms of sampled broilers. Our findings also suggest, however, that other bacterial and viral infectious agents not investigated in this study may be involved in the etiology of the respiratory problems of these birds, since those that... (Complete abstract click electronic access below) / Mestre

This dissertation is an investigation into the effect of different zinc sources and levels on eggshell quality and microstructure, as well as keel bone damage. Eggshell function is two-fold; eggshells function to protect the developing embryo, as well as act as a barrier against bacterial penetration, optimizing food safety of the egg for human consumption (Mabe et. Al., 2003). Two small trials were conducted in order to determine differences in eggshell microstructure of eggs produced from hens at peak lay (26 weeks of age) and at the end of lay (88 weeks of age). Two groups of hens were fed a calcium sufficient or a calcium deficient diet. From this it was determined that eggs with higher breaking strengths had some differences in eggshell microstructure. Eggs with greater breaking strengths had a greater density of ‘normal’ structures, compared to ‘abnormal’ structures. Additionally, eggs requiring a greater breaking force, had a thicker micro-structure, compared to shells requiring less breaking force. With this knowledge on microstructure, a larger, 36-week study was conducted using different zinc sources. Every four weeks, eggs were collected and standard egg quality measurements were taken and keel bones were scored. At the end of the study, keel bones were collected from randomly selected hens representing each treatment. Picture of these keel bones were taken and measurements were taken to determine type and degree of deformation, in comparison to scores taken on the live bird. Pens selected for keel bone analysis, were the same pens that eggs were taken for imaging by the scanning electron microscope, to determine eggshell microstructure. From this data, it was determined that egg quality differences were detected, as well as differences in eggshell microstructure. Additionally, keel bone scores progressively worsened throughout the 36-week long study, with type and degree of deformation differing depending on zinc source.

Estrogens seem to play a role in the locomotor activating effects of cocaine. Japanese quail provide a good model for hormonal manipulation as alterations of their photoperiod controls hormone levels. The current study aims to examine the role of early life photoperiod manipulation in cocaine-induced behavioral sensitization in quail. It was expected that if quail were raised on a short photoperiod, they would have a reduction in gonadal hormones and this reduction in hormones would affect the acquisition of cocaine-induced behavioral sensitization. Quail were raised on an 8L:16D or a 16L:8D light cycle. Following 2 days of habituation, quail were administered saline, 5 mg/kg, or 10 mg/kg cocaine for 10 days. Restricted photoperiods in early life were correlated to lower gonadal hormone levels in females and males. Male quail raised on the short-light cycle developed a sensitized response to 10 mg/kg cocaine. Female quail raised on the short- or long-photoperiod developed behavioral sensitization to 5 mg/kg cocaine. Furthermore, early life reduction in estradiol in females modulated the amount of activity on day 10 of cocaine treatment. The current research extends previous research by finding a possible early life gonadal hormone control of behavioral sensitization in the quail.

The objective of this thesis was to evaluate the effect of exogenous enzyme supplementation, phytase and xylanase-glucanase, on AME value of barley in poultry and swine. In the first study, 280 broilers were assigned 1 of 8 treatments. Barley inclusion in the diet resulted in decreased (P < 0.05) performance. There was a treatment × phytase × xylanase-glucanase interaction for dry matter retention with birds fed the corn-SBM-barley diet supplemented with phytase and xylanase-glucanase having higher (P < 0.05) DM retention compared to birds fed corn-SBM-based diet with only xylanase-glucanase supplementation. AME and AMEn of corn-SBM-based diets were greater (P < 0.05) than the corn-SBM-barley-based diets. Energy metabolizability and AMEn of barley significantly increased with xylanase-glucanase supplementation. In the second study, 24 pigs (12 pigs/phase) were assigned to 1 of 4 treatments with xylanase-glucanase and phytase. After a 7-d adaption period, urine and feces were quantitatively collected for 5 d. DE of the barley-based diet supplemented with xylanase-glucanase (3,578 kcal/kg) and phytase and xylanase-glucanase in combination (3,617 kcal/kg) were significantly different. Compared to control diets, exogenous enzymes either significantly improved or had a tendency to improve AME and AMEn value of barley in broilers, but not in growing pigs.

Feed accounts for the highest cost associated in poultry production, with energy-containing feedstuffs being the most expensive portion of the cost of feeding. The increasing demand for poultry meat gives reason to determine accurate apparent metabolizable energy (AME) values for various feedstuff through measuring energy utilization in the birds. The adaptation length of birds fed an experimental diet may affect the determined AME value due to the diet matrix and physiochemical properties of the feedstuff. Therefore, the objective of this thesis was to evaluate a select group of energy-containing feedstuff with different diets and with factors such as coccidia challenge and exogenous enzyme supplementation that may influence the determined AME values in broiler chickens.

Bird strikes ( collisions between birds and aircraft) pose a significant threat to aviation safety. For example , Naval Air Facility El Centro, California, lost an F-18 jet to a bird strike in October 1995. To help combat the bird-strike threat at Naval Air Facility El Centro, I developed a bird-avoidance model as a risk-management tool for the installation. It can be used to schedule flights at NAF El Centro and its two associated practice-bombing ranges during times of low-bird activity. I calculated bird-strike risks and published them in web-page format on both the installation's server and the USGS/Utah State University, College of Natural Resources' server for easy access by flight crews, flight-safety officers, airfield managers, natural resource managers, and other Navy personnel. Bird hazards during daylight hours were quantified using daily bird counts through the year 2000. These were combined with a bird-hazard index for various species, developed using U.S. Air Force bird-strike records. Nocturnal bird hazards were quantified in the fall of 2000 using a bird-radar system to count birds in three relative size classes. Large- and medium-sized birds were scaled to represent a higher risk to aircraft than small birds. Nocturnal bird hazards beyond the fall study were estimated using U.S. Air Force bird-strike records. The main section of the web page allows the user to select the area and time of year, which links to the appropriate color-coded bird-strike-risk graph. The graphs describe the bird-strike risk by time of day and altitude with red for high risk, yellow for moderate risk, and green for low risk. The web page also identifies and describes the most hazardous bird species in the area, recommends methods of hazard management, and provides links to bird-strike-information sources on the web.