Dissertations / Theses on the topic 'Psittacines'

The present report is based on the author’s experience throughout performed externships for the conclusion of the Integrated Master’s Degree in Veterinary Medicine by the University of Évora. It is composed of two parts: the first concerns accompanied clinical cases in different exotic species at Centro Veterinário de Exóticos do Porto (in Porto, Portugal); the second comprises a bibliographical review regarding the topic “Egg-Binding and Dystocia in Psittacines” and three clinical cases. The second part also includes a chapter regarding the author’s personal considerations where the differences between clinical practice of pet and captive birds are concerned, based on the experience at a zoological institution in the Canary Islands, Spain; Resumo: Clínica de Animais Exóticos O presente trabalho é baseado na experiência da autora nos estágios efetuados no âmbito da conclusão do Mestrado Integrado em Medicina Veterinária pela Universidade de Évora. Este trabalho é composto por duas partes: a primeira concerne os casos clínicos acompanhados em diversas espécies exóticas no Centro Veterinário de Exóticos do Porto (no Porto, Portugal); a segunda consiste numa revisão bibliográfica acerca de “Retenção de ovos e distócia em Psitacídeos”, assim como a apresentação de três casos clínicos. A segunda parte inclui também uma secção de considerações pessoais da autora relativas às diferenças entre a prática clínica de aves exóticas como animal de companhia e em cativeiro, baseadas na experiência de estágio numa instituição zoológica nas Ilhas Canárias, Espanha.

Para o presente estudo, utilizaram-se amostras de excretas cloacais de 65 aves da família Psittacidae, previamente sexadas. Os andrógenos e estrógenos fecais foram extraídos com Tampão Fosfato Salino (PBS) e com uma solução PBS:Álcool Etílico (4:1) e a mensuração hormonal foi realizada em ?kits? comerciais para radioimunoensaio no Laboratório de Dosagens Hormonais (LDH) do Departamento de Reprodução Animal (VRA) da Faculdade de Medicina Veterinária e Zootecnia (FMVZ) da Universidade de São Paulo (USP). O sexo de cada ave foi confirmado utilizando como parâmetro o intervalo de confiança (95%) da média dos valores transformados do fator Razão dos índices de Estrógenos e Andrógenos ou com o intervalo de confiança (95%) da média dos valores transformados dos índices do fator Andrógeno. As extrações de DNA de algumas amostras foram realizadas por meio de ?kits? comerciais e por meio de resinas no Laboratório de Biologia Molecular do Departamento de Patologia (VPT) - FMVZ/USP. O DNA foi amplificado pela técnica da PCR utilizando-se oligonucleotídeos iniciadores específicos para a região que determina o sexo das aves. Também se avaliou o efeito da adição de células sanguíneas nas amostras. Setenta por cento das aves tiveram o sexo confirmado pela técnica do radioimunoensaio. A amplificação do produto na PCR somente ocorreu em amostras onde células sanguíneas foram adicionadas e o DNA extraído com um determinado ?kit? comercial. Os resultados encontrados demonstram a necessidade da realização de mais estudos para a determinação do sexo de aves monomórficas por meio de técnicas não invasivas. Validaram-se dois kits comerciais para a mensuração de metabólitos de hormônios esteróides em excretas cloacais de aves por radioimunoensaio.<br>For the current study, it were used fecal samples from 65 birds of Psittacine Family, previously sexed by PCR from blood cells. The fecal androgens and estrogens metabolites were extracted with PBS (Phosfate Buffer Saline) or PBS :Ethil Alchool (4:1) and measured by comercial radioimunoassay kits at the Hormonal Measurement Laboratory of the Department of Animal Reproduction of the Faculty of Veterinary Medicine and Zootechny of the University of São Paulo. The sex determination of the birds were performed using the confidence interval (95%) for the mean of transformed values of estrogens/androgen rate or transformed androgens values. The fecal DNA extraction of some samples were performed using comercial kits or by resines at the Molecular Biology Laboratory of the Pathology Department of the Faculty of Veterinary Medicine and Zootechny of the University of São Paulo. The DNA were amplified by PCR using specific primers for the target region of bird gene that express sex. Evaluation of the effect of blood cells adding on samples also had been done. 70% of birds had the sex confirmed by radioimunoassay. The amplification of PCR products occured only in samples added with blood cells and extracted by a specific comercial kit. The results showed that further studies for sex determination on monomorphic birds by non-invasive techniques are necessary. Two comercial kits were validated for the measurement of fecal steroid metabolites by radioimunoassay.

Este estudo retrospectivo revisa os achados radiográficos em exames simples e contrastados de 38 aves psittaciformes com alterações no proventrículo. Os dados foram obtidos do Serviço de Diagnóstico por imagem do departamento de cirurgia junto ao Hospital Veterinário da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, durante o período de janeiro de 2004 a abril de 2012. A distribuição das alterações, assim como as suas respectivas espécies e idades, foram estudadas. O sexo das aves não foi considerado neste estudo. A espécie Amazona aestiva (papagaio-verdadeiro) alcançou a porcentagem mais elevada (44,8%) entre as diversas ordens, seguidos pelos Nymphicus hollandicus (Calopsita) (28,9%). Neste período de oito anos, os sintomas mais comuns observados na anamnese foram a \"apatia/prostração\" e \"anorexia/prostração\" (52,63% cada). Entre as suspeitas clínicas mais frequentes, a doença da dilatação do proventrículo (PDD) foi a mais citada (26,32%). O achado radiográfico mais frequentemente observado foi a \"dilatação do proventrículo por conteúdo de radiodensidade gás e líquido\" (34,21%). A medição do proventrículo foi um dado descrito em 34,84% dos laudos. Dentre as aves estudadas com alterações radiográficas em proventrículo, o exame radiográfico contrastado por sulfato de bário foi solicitado em 26,32% dos casos, onde o achado radiográfico mais comum foi a \"retenção do meio de contraste ou evolução lenta do mesmo\". Dentre os casos com suspeita clínica de PDD, os únicos nos quais foi possível obter confirmação foram aqueles em que foi solicitado o exame contrastado. Algumas aves também exibiram alterações radiográficas inespecíficas. A dilatação de proventrículo em si não é patognomônica para PDD, visto que esta pode vir de forma atípica e que há diversas outras doenças que causam alterações semelhantes.<br>This retrospective study examines radiographic findings of plain and contrastenhanced examinations of 38 psittacine birds presenting proventricular abnormalities. The data were obtained by the Image Diagnostic Service of the Veterinary Hospital of the School of Veterinary Medicine and Animal Science of the University of São Paulo between January 2004 and April 2012. The distribution of the abnormalities, as well as the birds species and ages, were also investigated. The birds sex was not taken into account. The highest percentage among the different orders (44.8%) was observed among specimens of Amazona aestiva (Blue-fronted Parrot), followed by Nymphicus hollandicus (Cockatiel) (28.9%). In the eight-year period of the study, the most common symptoms were \"apathy/weakness\" and \"anorexia/weakness\" (52.63% each.) Proventricular Dilatation Disease (PDD) was the most frequent clinical suspicion (26.32%.) The most commonly observed radiographic abnormality was a \"Proventricular dilatation caused by content of gas and water radiodensity\" (34.21%.) The measurements of the proventriculus were described in 34.84% of all reports. Among the birds studied with radiographic alterations in proventriculus, radiographic examination contrasted by barium sulfate was required in 26.32% of cases. The most common findings were \"retention or slow transit of the contrast agent\". The only cases with a clinical suspicion of PDD for which it was possible to obtain a confirmation were those for which a contrast-enhanced examination was ordered. Some birds also presented radiographic abnormalities of uncertain diagnosis. A proventricular dilatation in itself is not pathognomonic of PDD inasmuch as said disease may present in an atypical manner and several other diseases may cause similar abnormalities.

Os fatores de virulência presentes em Escherichia coli (E. coli) patogênicas vêm sendo estudados em diversas espécies animais devido à importância de algumas cepas. Estas podem ser classificadas em sorotipos ou mais recentemente através de patotipos, de acordo com algumas técnicas de biologia molecular. Os patotipos mais conhecidos atualmente incluem as ETEC, STEC, EPEC, EIEC, UPEC e APEC. Vários patotipos encontrados causando doenças em animais têm também afetado gravemente seres humanos. Diversas espécies de aves também revelaram possuir patotipos de E. coli capazes de causar infecções zoonóticas. Além do aspecto zoonótico, infecções por bactérias Gram-negativas em aves silvestres têm especial importância, pois o Taxon Psittacidae contém muitas das espécies que mais estão ameaçadas de extinção, e sua manutenção e reprodução em cativeiro é comprometida por infecções bacterianas constantes, em especial por E. coli. Nesse estudo 174 amostras de swabs cloacais de aves sintomáticas e assintomáticas e de casos de necropsias foram testadas através da técnica da PCR para a detecção de fatores de virulência e a classificação em grupos filogenéticos. Detectaram-se 93 amostras (53.45%) positivas para associações de fatores de virulência, em especial para os patotipos EPEC, APEC e UPEC e a classificação de grande parte das amostras de necropsias no grupo filogenético A. O gene iss foi encontrado com maior freqüência, sendo esta de 51.7% (n=30) nas aves sintomáticas e 23.2% (n=23) naquelas assintomáticas. Os resultados demonstraram serem de interesse não apenas com relação ao potencial zoonótico dessas aves em cativeiro, mas também para programas de conservação que visem à liberação de aves no meio selvagem.<br>Virulence factors of pathogenic Escherichia coli (E.coli) have been studied in several animal species due the importance of certain strains. These strains can be classified in serotypes and more recently in pathotypes using molecular biology techniques. The mostly known pathotypes nowadays include ETEC, STEC, EPEC, EIEC, UPEC and APEC. Several pathotypes causing disease and found in animals are also linked to severe disease in humans. A number of bird species are also known to possess pathotypes able to cause zoonotic infections. Besides de zoonotic aspect, infections by Gram-negative bacteria have a special importance as the Taxon Psittacidae includes many of the most threatened species and their maintenance in captivity is compromised by constant bacterial infections, especially E.coli. In this study, 174 samples from cloacal swabs of asymptomatic and symptomatic birds and necropsy cases were tested employing the PCR technique to detect virulence factors and to classify among phylogenetic grups. Ninety three positive samples were detected for virulence factors associations, mostly EPEC, APEC and UPEC, and most of the necropsy samples for the phylogenetic group A. The iss gene was detected with a higher frequency in symptomatic birds (51.7%, n=30) and 23.2%, (n=23) in asymptomatic ones. The results reveal to be of interest not only to the zoonotic potential of the isolates but also for conservation programs that intend to release these birds into the wild.

Os psitacídeos estão entre as espécies de aves mais apreendidas e encaminhadas aos centros de triagem animal em São Paulo. Também são comumente mantidos em ambiente doméstico como aves de estimação. A manutenção destas aves em cativeiro pode representar um risco zoonótico e contribuir para a propagação das estirpes de enterobactérias multirresistentes, como Klebsiella spp. produtora de beta-lactamase de espectro estendido (ESBLs), que podem interferir no tratamento de infecções nosocomiais em humanos. O objetivo deste estudo foi identificar e caracterizar estirpes de Klebsiella spp. isoladas de secreções respiratórias de 46 psitácideos doentes, determinando a virulência e o perfil de resistência a 15 antimicrobianos. Dentre as 19 estirpes de Klebsiella spp. isoladas, 16 (16/19) foram identificadas como Klebsiella pneumoniae, e três (3/19) foram identificadas como Klebsiella oxytoca. O perfil de suscetibilidade aos antimicrobianos demonstrou alta resistência para ampicilina (89,5%), e o perfil de virulência demonstrou uma alta prevalência dos genes fimH (94,7%), kpn (89,4%), uge (84,2%) e irp-2 (78,9%). Três estirpes de K. pneumoniae foram positivas para produção de beta-lactamase de espectro estendido. Estas estirpes foram classificadas nos sequence types (STs) ST15, ST147 e ST307. Esses três grupos clonais representam os principais responsáveis por surtos de infecções hospitalares por K. pneumoniae no mundo. No entanto, esse é o primeiro relato desses clones como causadores de doença em aves. Esses dados indicam a ocorrência de K. pneumoniae produtora de CTX-M-15 e CTX-M-8 em psitacídeos cativos e confirmam o potencial zoonótico e antropozoonótico do agente, destacando a relevância clínica para humanos e animais.<br>Psittacine birds are among the most seized bird species that are sent to animal sorting centers in São Paulo. They are also commonly kept in the domestic environment like pet birds. The maintenance of these birds in captivity may represent a zoonotic risk and contribute to the propagation of strains of multiresistant enterobacteria, such as Klebsiella spp. beta-lactamase extended-spectrum (ESBLs), which may interfere in the treatment of nosocomial infections in humans. The objective of this study was to identify and characterize strains of Klebsiella spp. isolated from respiratory secretions of 46 diseased psittacines, determining virulence and resistance profile to 15 antimicrobials. Among the 19 strains of Klebsiella spp. isolated, 16 (16/19) were identified as Klebsiella pneumoniae, and three (3/19) were identified as Klebsiella oxytoca. The antimicrobial susceptibility profile demonstrated high resistance to ampicillin (89.5%), and the virulence profile demonstrated a high prevalence of fimH (94.7%), kpn (89.4%), uge (84.2% %) and irp-2 (78.9%). Three strains of K. pneumoniae were positive for extended-spectrum beta-lactamase production. These strains were classified in sequence types (STs) ST15, ST147 and ST307. These three clonal groups represent the main responders for outbreaks of K. pneumoniae nosocomial infections worldwide. However, this is the first account of these clones as causing disease in birds. These data indicate the occurrence of K. pneumoniae producing CTX-M-15 and CTX-M-8 in captive parrots and confirm the zoonotic and anthropozoonotic potential of the agent, highlighting the clinical relevance for humans and animals.

O presente trabalho visa relatar a casuística observada durante o estágio curricular em clínica de animais exóticos e silvestres, no Centro Veterinário de Exóticos do Porto e no Parc Zoologique d’Amnéville. Esta será exposta em frequências absolutas e relativas que estarão representadas em gráficos e tabelas para que a interpretação das atividades desenvolvidas seja clara. De seguida, estará contemplada uma revisão bibliográfica subordinada a uma das emergências mais frequentemente assistidas - as patologias relacionadas com o mau maneio nutricional em psitacídeos de cativeiro. Após uma breve caracterização dos elementos da ordem Psittaciforme, segue-se a descrição da sua morfologia e fisiologia gastrointestinal, e uma revisão da sua alimentação na natureza. As patologias nutricionais mais comuns serão referenciadas, assim como a alimentação atualmente recomendada, a qual será nutricionalmente comparada as misturas de sementes. Por fim, será apresentado um caso clínico de um animal acompanhado durante o estágio, acometido pela lipidose hepática; Abstract: “Internship report on clinic of exotic and feral animals - nutritional pathologies in psittacines” The following work describes the activities performed during the internship on clinic of exotic and feral animals at Centro Veterinário de Exóticos do Porto and Parc Zoologique d'Amnéville. These will be displayed in absolute and relative frequencies, which, in turn, will be represented in graphics and tables to allow a clear interpretation of the developed activities. Then, a literature review will be made of the most frequently assisted emergencies - the pathologies related to a poor nutritional management in captive psittacines. A brief characterization of some elements of the Psittaciforme order will be followed by a description of their gastrointestinal morphology and physiology, and by a review of their food habits in the wild. The most common nutritional disorders will be referred, as well as the currently recommended diet, which will be nutritionally compared with seed mixtures. Finally, it will be presented a clinical case of an animal accompanied during the internship, suffering from hepatic lipidosis.

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Agradecemos a compreensão. on 2017-01-20T15:03:30Z (GMT)<br>Submitted by Ludmilla Geraldo Di Santo (ludmilladisanto@gmail.com) on 2017-03-09T18:57:40Z No. of bitstreams: 1 Dissertação - Ludmilla Geraldo Di Santo.pdf: 3261093 bytes, checksum: d46f60bc493c74390b3bc14fee0f382f (MD5)<br>Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-03-20T19:30:16Z (GMT) No. of bitstreams: 1 disanto_lg_me_jabo.pdf: 3261093 bytes, checksum: d46f60bc493c74390b3bc14fee0f382f (MD5)<br>Made available in DSpace on 2017-03-20T19:30:16Z (GMT). No. of bitstreams: 1 disanto_lg_me_jabo.pdf: 3261093 bytes, checksum: d46f60bc493c74390b3bc14fee0f382f (MD5) Previous issue date: 2016-12-05<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)<br>Estudos sobre processamento de rações e sua influência no metabolismo são escassos para psitacídeos. Desta forma, o presente estudo teve como objetivo avaliar o consumo, digestibilidade, parâmetros bioquímicos, hematológicos e radiográficos de papagaios-verdadeiro após a transição de dieta com alta gordura, baseada em girassol, para alimento balanceado para manutenção de papagaios. Esta formulação balanceada foi processada de modo a se obter três diferentes graus de cozimento do amido: Ração peletizada (PEL) - ingredientes moídos com peneira de 2mm e peletizados, com 27% de cozimento do amido; Ração extrusada baixo cozimento (EXTb) - ingredientes moídos com peneira de 2mm e extrusados com baixa transferência de energia, com 82% de cozimento do amido; Ração extrusada elevado cozimento (EXTa) - ingredientes moídos com peneira de 0,5mm e extrusados com elevada transferência de energia, com 98% de cozimento do amido. Trinta papagaios-verdadeiro adultos foram mantidos na dieta à base de girassol por 90 dias, para homogeneização do grupo e determinação dos valores basais. Posteriormente foram distribuídos em delineamento inteiramente casualizado com 10 repetições (papagaios) por tratamento (rações experimentais) e mantidos nas três dietas por um período de 160 dias. Foram determinados o consumo e a palatabilidade dos alimentos, coeficiente de digestibilidade aparente dos nutrientes, concentrações de ácidos graxos voláteis - AGVs (acético, butírico, propiônico, valérico, isovalérico e isobutírico), lactato e amônia das excretas. Ao início e final do experimento foram avaliados os parâmetros hematológicos, radiográficos e as concentrações plasmáticas de aspartato aminotransferase (AST), albumina, colesterol, triglicérides, glicose e ácido úrico. Os dados foram submetidos a análise de variância e as médias comparadas pelo teste de Tukey (P<0,05). A dieta PEL apresentou maior digestibilidade da gordura e do amido que a EXTa e EXTb (P<0,05), bem como tendência à produção de excretas com maior concentração de ácido acético e ácidos graxos de cadeia curta totais (P<0,1). Por sua vez, o girassol apresentou maior digestibilidade da matéria seca (81,3%), matéria orgânica (87,4%), extrato etéreo (97,5%), energia bruta (87,3%) e tendência de maior digestibilidade da proteína bruta (84,0%) quando comparada à médias das rações processadas (matéria seca – 67,9%, matéria orgânica – 76,7%, extrato etéreo – 89,9%, energia bruta – 74,6%, proteína bruta – 77,6%) (P<0,05). A transição de uma dieta com elevada gordura (girassol) para ração balanceada promoveu melhor perfil bioquímico sérico, com redução de glicose (242,7 para 216,0 mg/dl), triglicérides (163,9 para 111,1 mg/dl), colesterol (263,6 para 184,1 mg/dl) e AST (190,8 para 113,5 U/L) (P<0,05). O melhor aporte nutricional das rações experimentais proporcionou melhora nos parâmetros hematológicos e imunocompetência dos papagaios, com maiores concentrações de hemácias, hemoglobina, hemoglobina corpuscular média, leucócitos totais, linfócitos e monócitos, quando comparado aos valores mediante ingestão do girassol (p<0,05). Ao exame radiográfico os papagaios apresentaram redução na largura da ampulheta (coração – fígado) e na relação coração fígado (p < 0,05) após o consumo da ração processada. Foi verificado que o cozimento do amido não modificou o metabolismo dos papagaios, mas a transição de dieta com elevado teor de gordura (54,4%), como o girassol, para as rações balanceadas melhorou o metabolismo reduzindo as concentrações séricas de glicose, triglicérides, colesterol e enzima hepática, além de promover aumento de hemácias e linfócitos.<br>Studies on food processing and its influence on metabolism are scarce for psittacines. The present study evaluated the consumption, digestibility, serum biochemistry, and radiographic parameters of blue-fronted amazon parrots after the transition from high-fat diet, based on sunflower seed, to a balanced formulation for parrot maintenance. This formulation was processed to obtain three different degrees of starch gelatinization: Pelletized food (PEL) - ingredients ground with a 2mm sieve and pelletized, with 27% of starch cooking; Low cooking extruded feed (EXTL) - ingredients ground with 2 mm sieve and extruded with low energy transference, with 82% of starch cooking; High cooking extruded food (EXTH) - ingredients ground with 0.5 mm sieve and extruded with high energy transference, with 98% of starch cooking. Thirty adults blue-fronted amazon parrots were maintained on the sunflower seed based diet for 90 days, for homogenization of the group and determination of baseline values. Later, they were distributed in a completely randomized design with 10 repetition (parrots) per treatment (experimental food) and fed with the diets for a period of 160 days. Food consumption and palatability, apparent digestibility coefficient of nutrients, concentrations of volatile fatty acids (VFAs - acetic, butyric, propionic, valeric, isovaleric and isobutyric), lactate and ammonia of the excreta and, at the beginning and at the end of the experiment hematological, radiographic and plasma concentrations of aspartate aminotransferase (AST), albumin, cholesterol, triglycerides, glucose and uric acid were determined. The data were submitted to analysis of variance and the means compared by the Tukey's test (P <0.05). The PEL food presented higher fat and starch digestibility than the EXTL and EXTH (P<0.05) and a tendency for higher excreta concentrations of acetic acid and total short chain fatty acids (P<0.1). On the other hand, sunflower seed presented higher digestibility of dry matter (81.3%), organic matter (87.4%), fat (97.5%), crude energy (87.3%), and crude protein (84.0%) than the mean of the formulated foods (P<0.05; dry matter - 67.9%, organic matter - 76.7%, fat - 89.9%, crude energy - 74.6%, crude protein - 77.6%). The transition from a high fat diet (sunflower seed) to a balanced food improved the serum biochemical profile, reducing the concentrations of glucose (from 242.7 to 216.9 mg/dl), triglycerides (from 163.9 to 111.1 mg/dl), cholesterol (from 263.6 to 184.1 mg/dl) and AST (from 190.8 to 113.6 U/L)(P<0.05). The better nutrient profile of the formulated foods improved blood parameters and immunocompetence, with higher concentrations of hemoglobin, mean corpuscular hemoglobin, total leukocytes, lymphocytes and monocytes than when the parrots were fed with sunflower seeds (P<0.05). The radiographic exam showed a reduction in the width of the hourglass (heart - liver) and heart - liver ratio after the intake of the formulated foods (P<0.05). Food processing did not modify the metabolism of parrots, but the transition from a high-fat diet (54.4%), such as sunflower seeds, to balanced diets improved parrot metabolism reducing serum glucose, triglycerides, cholesterol and liver enzyme concentrations, improving the production of red cells and lymphocytes.

No Brasil, o papagaio-verdadeiro (Amazona aestiva) é uma das aves mais procuradas como animal de estimação e comercializadas ilegalmente. Na literatura pouco é descrito sobre a microbiota intestinal de aves silvestres. O trato intestinal das aves é composto por inúmeras e diferentes espécies bacterianas. A grande maioria são bactérias gram-positivas pertencentes ao grupo de bactérias ácido-láticas. Este estudo teve como objetivo isolar e identificar a presença de bactérias dos gêneros Lactobacillus, Bifidobacterium, Enterococcus, Pediococcus e Lactococcus na microbiota entérica de papagaios Amazona aestiva de vida livre e de cativeiro. Para isto foram coletadas amostras de 26 aves de vida livre e de 26 aves procedentes de dois criadouros comerciais. O Enterococcus foi o gênero que apresentou maior freqüência de isolamentos (100%), seguido dos gêneros Pediococcus (63,46%), Lactobacillus (28,84%), Lactococcus e Bifidobacterium (15,38%). Foram isoladas 12 espécies de Enterococcus, sendo o E. faecium a espécie que apresentou maior ocorrência de isolamento, presente em 63,46% das aves, seguido por E. faecalis isolado em 57,69% das aves, Enterococcus sp. identificado em 46,15% das aves, E. hirae em 30,76% e E. raffinosus em 19,23%. Seis espécies de Pediococcus foram isoladas, sendo que P. pentosaceus foi a mais freqüente e esteve presente em 57,69% das aves. Foram isoladas cinco (5) espécies de Lactococcus, sendo L. lactis subsp. cremoris isolados em 3,84% das aves e Lactococcus sp. em 9,61%. Lactobacillus apresentou uma maior diversidade, com 14 espécies identificadas, sendo as mais freqüentes L. coryniformis subsp. torquens e L. sanfrancisco com 7,69% de aves positivas para cada espécie. Três (3) espécies de Bifidobacterium foram isoladas, sendo B. bifidum identificado em 9,61% das aves. Estudos complementares precisam ser conduzidos para uma melhor compreensão da microbiota intestinal das aves silvestres, assim como analisar as similaridades e diferenças com as aves domésticas, o que permitirá um manejo apropriado e menos empírico desta espécie em cativeiro.<br>In Brazil, Blue-fronted Parrot (Amazona aestiva) has been widely owned as a pet bird and, therefore, one of the Brazilians birds most frequently traded illegally in the Black Market. There are few reports in the current literature regarding to the microbiota of wild birds. The gastrointestinal tract of these birds has a wide variety of bacterial species; most of them are Gram positive bacteria and belongs to the lactic acid group. The present study has isolated and identified Lactobacillus, Bifidobacterium, Enterococcus, Pediococcus, and Lactococcus bacterias present in fecal samples of wild and captive Amazona aestiva parrots. Fifty two fecal samples were collected from 26 wild parrots and 26 parrots from commercial breeders. Enterococcus genus was the most frequently isolated (100%), followed by Pediococcus (63.46%), Lactobacillus (28.84%), Lactococcus and Bifidobacterium (15.38%). Twelve species of Enterococcus were identified. E. faecium was the most frequently isolated from the birds representing 63.46%, followed by E. faecalis (57.69%), Enterococcus sp. (46.15%), E. hirae (30.76%), and E. raffinosus (19.23%). P. pentosaceus was identified from 57.69% of the parrots. This specie was the most frequently isolated. Five different species of Lactococcus were found out. Lactococcus sp. was identified from 9.61% of the birds, while L. lactis subsp. lactis represented 3.84%. Fourteen different species of Lactobacillus were isolated, showing the biggest diversity among all the studied genera. L. coryniformis subsp. torquens and L. sanfrancisco were isolated from 7.69% of the birds. Three different species of Bifidobacterium were isolated, and B. bifidum was identified in 9.61% of the birds, being the most frequently isolated. Further studies are needed to a better comprehension of the microbiota in wild birds. Besides comparing differences and similarities between wildlife parrots and pet birds will allow appropriate and less empiric management of those birds in captivity.

Durante as últimas duas décadas, as técnicas de análise de esteroides fecais têm sido utilizadas para pesquisa em endocrinologia reprodutiva em diversas espécies animais silvestres de vida livre, ou cativeiro. O presente estudo propõe a validação da técnica do ensaio imunoenzimático antiprogestinas CL425 e antiandrógenos R156/7 fecais em papagaio-de-peito-roxo (Amazona vinacea). Objetiva-se descrever os perfis endócrinos anuais dos esteroides sexuais urofecais em Amazona vinacea com potencial aplicação para o aprimoramento do manejo reprodutivo da espécie. Foram utilizados 10 casais adultos da espécie Amazona vinacea mantidos em viveiros suspensos, em Lomba Grande, Rio Grande do Sul, Brasil. As excretas foram coletadas duas vezes por semana, entre maio de 2012 e maio de 2013. O monitoramento da atividade gonadal foi feito por mensuração de metabólitos de andrógenos nas excretas dos machos e progestágenos, nas de fêmeas. Foram coletadas amostras frescas de excretas, consistentemente, entre 12h00min e 16h00min e mantidas congeladas até o processamento. O perfil anual de andrógenos nas excretas de machos de A. vinacea apresentou médias mensais variáveis de 25,9 ng/g, em janeiro até 125,7 ng/g, em setembro. O perfil anual de progestinas nas excretas de fêmeas A. vinacea, as médias mensais variaram de 2,76 ng/g, em dezembro até 45,7 ng/g, em setembro. Os perfis reprodutivos apresentaram variação na atividade hormonal, durante a fase reprodutiva. Porém, comparando-se casais reprodutores com não-reprodutores, observa-se que os níveis de progestágenos não diferiram significativamente entre fêmeas que fizeram postura e aquelas que não fizeram. Entretanto, os níveis de andrógenos diferiram significativamente entre reprodutores e não-reprodutores. Diferenças hormonais foram observadas nas diferentes estações anuais, tanto nos machos, quanto nas fêmeas. Dados metereológicos de temperatura e insolação demostraram correlação negativa com os perfís anuais de progestágenos. A utilização da metodologia aqui mencionada demonstrou aplicabilidade, praticidade e segurança como ferramenta para o estudo da fisiologia e do manejo reprodutivos de Amazona vinacea.<br>During the last two decades, the techniques of analysis of fecal steroids have been applied for studying reproductive profiles in many species of captive wild and free living wild animals. The advantages of this methodology are widely understood. This study aimed to validate the anti- CL425 progestin and anti-R156/7 androgen enzyme immunoassay in fecal samples from parrot-breasted-purple (Amazon vinacea). This communication describes the annual endocrine profiles of urofecal sex steroids in Amazona vinacea with potential application for improving the species reproductive management. In total, 10 adult A. vinacea pairs kept in suspended aviaries in southern Brazil were studied. Excreta samples were collected twice weekly between May 2012 and May 2013. Gonadal activity was monitored by measurement of androgen and progestin metabolites in excreta of birds. Fresh excreta samples were collected always between 12:00 and 16:00 hs, and kept frozen until processing. The samples were dried at 57oC, hormones crushed and extracted using 80% methanol. The hormone dosage was performed at the Laboratory of Endocrinology, NUPECCE (Center for Research and Conservation of Deer, Jaboticabal, SP) using enzyme immunoassay with antibody to androgen and progestin. Feces from 5 A. vinacea pairs were used in two distinct periods for assay validation, within and outside the breeding season. Tests included Parallelism, Dose-Response, and Physiological Validation. It was concluded that the enzyme immunoassay for determining progestin and androgen profiles in urofecal samples from A. vinacea is accurate, precise and reliable. The annual profile of androgen in excreta of A. vinacea resulted the following monthly averages: January 25.9 ± 3.7 ng/g (n = 90); February 28.7 ± 3.2 ng/g (n = 60); March 31 7 ± 17.7 ng/g (n = 80); April 27.9 ± 5.5 ng/g (n = 90); May 43.3 ± 7.9 ng/g (n = 90); June 49.1 ± 15.9 ng/g (n = 80); July 36.5 ± 7.5 ng/g (n = 90); August 86.9 ± 46.2 ng/g (n = 90); September 42 125.7 ± 1 ng/g (n = 80); October 110.3 ± 63.2 ng/g (n = 90); November 74.2 ± 57.7 ng/g (n = 90) and December 43.3 ± 13 4 ng/g (n = 80). The monthly averages in the annual profile of progestin in excreta of A. vinacea were as follows: January 7.2 ± 0.9 ng/g (n = 90); February 9.26 ± 1.3 ng/g (n = 60); March 15 8 ± 2.9 ng/g (n = 80); April 14.15 ± 1.85 ng/g (n = 90); May 16.05 ± 3.16 ng/g (n = 90); June 11.85 ± 2.54 ng/g (n = 80); July 11.89 ± 2.17 ng/g (n = 90); August 13.52 ± 3.48 ng/g (n = 90); September 55 ± 45.7 ng/g (n = 80); October 15.44 ± 24.91 ng/g (n = 90); November 4.27 ± 6.42 ng/g (n = 90) and December 2.76 ± 0.42 ng/g (n = 80). Reproductive profiles showed variation in hormonal activity during the reproductive phase. When comparing breeding to non-breeding pairs, it was observed that the progestin levels did not differ significantly between laying and not-laying birds. However, the androgen levels differed significantly between breeding and non-breeding birds. In addition, hormonal differences were observed in both male and female birds in accordance with different annual seasons. Meteorological temperature and insolation data showed a negative correlation with the annual progestin profiles. Measurement of androgen and progestin metabolites in A. vinacea feces proved to be a practical and safe tool for studying reproductive physiology and management of the species.

By examining psittacine speech (primarily from a Grey parrot, Psittacus erithacus, named Alex) in a series of two acoustic studies, two articulatory studies and one perceptual study, this dissertation demonstrates that some aspects of human language are not unique to our species. The first two studies identify frequency, intensity and durational aspects of, respectively, Alex's vowels (/i,I,e,ε,æ,ɒ,ə,o,U,u/) and stop consonants (/p,b,t,d,k,g/) that either differ from or resemble their human counterparts (primarily I. M. Pepperberg). Our results indicate that Alex produces acoustically distinct phonemes using more high frequency information than do humans. In both studies, we use acoustic data to make predictions about articulation. We also compare Alex's speech to that of a mynah, and conclude that these species use different mechanisms to produce speech. The third and fourth study examine vowel articulation: The third study, an X-ray videotape analysis, establishes that Alex configures his vocal tract in distinct ways for /i/ and /ɒ/. The fourth study models more than 2800 Grey parrot vocal tract shapes as conjoined tubes with known area functions and calculates associated vowel formants. The relationship between these mathematical models and formant values for /i,I,e,ε,æ,ɒ,ə,o,U,u/ is consistent with findings from the vowel study, X-ray study, Grese's unpubl. data and personal observations of relationships between Alex's vocal tract configurations and vowel production. The third and fourth studies show that Alex's "phonemes" are articulatorily distinct. The fifth study is perceptual and examines the relationship between acoustic characteristics of psittacine vowels and the accuracy with which listeners perceive them. We find evidence that Alex's acoustically least "human" vowel, /i/, is very difficult for listeners to perceive unless they have substantial exposure to Alex (on the order of several months). More acoustically prototypical vowels, like /ɒ/, are accurately identified even by less experienced listeners. We thus find that, at least for experienced listeners, Alex's "phonemes" are perceptually distinct. In sum, the dissertation provides acoustic, articulatory and perceptual evidence of phonemes and other phonetic structure in the speech of a psittacid.

To enable assessment of recombinant BFDV capsid protein (recBFDVcap) for vaccination to protect against PBFD, commercially available lovebirds (Agapornis sp.) were tested for evidence of past and current BFDV infection using PCR, HI and HA to identify suitable BFDV-free birds in which to test the vaccine. During this attempt, it was found that lovebirds from commercial aviaries were endemically infected with BFDV with evidence of up to 100% prevalence of BFDV DNA in blood samples from individual birds over time. Such an approach was abandoned as unlikely to yield suitable numbers of naïve birds to conduct a BFDV vaccination trial. As commercially available lovebirds were considered to be a poor source of BFDV-free birds, wild caught cockatoo nestlings and eggs (long-billed corella; Cacatua tenuirostris and galah; Eolophus roseicapillus) were used to assess the efficacy of BFDV vaccination using baculovirus recombinant BFDV capsid. Eggs were artificially incubated and 3 eggs successfully hatched and 1 was successfully hand-reared. All nestlings were screened for BFDV DNA in blood using PCR upon arrival then on days 11, 18 and 25 and tested for anti-BFDV antibody on the day of arrival. All hatched birds were determined to be free of BFDV DNA and BFDV HI antibody in the peripheral blood throughout the hand rearing period and the flock was considered to be suitable for a BFDV vaccination trial. Corellas (n=13) were injected with 1 mL of vaccine containing 10 μg recBFDVcap on day 0 and 0.4 mL vaccine containing 66.8 μg recBFDVcap on day 11. All vaccinated corellas and 5 non-vaccinated control corellas were given 0.4 mL BFDV suspension (titre = log2 12 HAU/50 μL) intramuscularly and 0.1 mL orally 16 days after booster vaccination. Blood was collected periodically during the vaccination period and blood and feathers collected before and after BFDV administration. Testing included BFDV DNA detection by PCR and qRT PCR (on blood) as well as serum antibody detection by haemagglutination inhibition (HI) and BFDV DNA and antigen was detected by qRT PCR and haemagglutination (HA) (on feathers), respectively. Four of 97 blood samples collected from vaccinated birds post BFDV challenge tested positive by PCR, whereas 17 of 35 samples taken from non-vaccinated control corellas tested positive. Vaccinated birds did not develop feather lesions, had only transient PCR detectable viraemia and had no evidence of persistent infection 270 days post-challenge using PCR, histopathology and immunohistochemistry (IHC). Non-vaccinated control corellas developed transient feather lesions and PCR, HI and HA test results consistent with PBFD. They were BFDV PCR positive for up to 41 days post-challenge and qRT PCR demonstrated reduced virus replication in vaccinated birds compared to non-vaccinated control birds. Thus, administration of recBFDVcap vaccine alone was found to incite an adaptive immune response in BFDV-free corellas that subsequently conferred protection against inoculation with BFDV. A commonly utilized method for excising blood dried on filter paper was proven to be of high risk of carryover contamination facilitated by a hole punch used for processing several samples. Therefore a practical method of avoiding carryover contamination was developed and used in the DNA testing procedures of the vaccination trial. Finally, the haematological characteristics of the above mentioned cockatoos were studied before and for 97 days after experimental infection with BFDV. It was found that the pre-challenge haematological values were similar between the vaccinated and non-vaccinated corellas. Most pre-challenge parameters were comparable to previously reported values of other cockatoos and psittacine birds. Significant differences were seen in both groups when comparing pre-challenge values with post challenge values for total and differential leukocyte concentrations, but PCV and TSP were not significantly affected by BFDV challenge.

To enable assessment of recombinant BFDV capsid protein (recBFDVcap) for vaccination to protect against PBFD, commercially available lovebirds (Agapornis sp.) were tested for evidence of past and current BFDV infection using PCR, HI and HA to identify suitable BFDV-free birds in which to test the vaccine. During this attempt, it was found that lovebirds from commercial aviaries were endemically infected with BFDV with evidence of up to 100% prevalence of BFDV DNA in blood samples from individual birds over time. Such an approach was abandoned as unlikely to yield suitable numbers of naïve birds to conduct a BFDV vaccination trial. As commercially available lovebirds were considered to be a poor source of BFDV-free birds, wild caught cockatoo nestlings and eggs (long-billed corella; Cacatua tenuirostris and galah; Eolophus roseicapillus) were used to assess the efficacy of BFDV vaccination using baculovirus recombinant BFDV capsid. Eggs were artificially incubated and 3 eggs successfully hatched and 1 was successfully hand-reared. All nestlings were screened for BFDV DNA in blood using PCR upon arrival then on days 11, 18 and 25 and tested for anti-BFDV antibody on the day of arrival. All hatched birds were determined to be free of BFDV DNA and BFDV HI antibody in the peripheral blood throughout the hand rearing period and the flock was considered to be suitable for a BFDV vaccination trial. Corellas (n=13) were injected with 1 mL of vaccine containing 10 μg recBFDVcap on day 0 and 0.4 mL vaccine containing 66.8 μg recBFDVcap on day 11. All vaccinated corellas and 5 non-vaccinated control corellas were given 0.4 mL BFDV suspension (titre = log2 12 HAU/50 μL) intramuscularly and 0.1 mL orally 16 days after booster vaccination. Blood was collected periodically during the vaccination period and blood and feathers collected before and after BFDV administration. Testing included BFDV DNA detection by PCR and qRT PCR (on blood) as well as serum antibody detection by haemagglutination inhibition (HI) and BFDV DNA and antigen was detected by qRT PCR and haemagglutination (HA) (on feathers), respectively. Four of 97 blood samples collected from vaccinated birds post BFDV challenge tested positive by PCR, whereas 17 of 35 samples taken from non-vaccinated control corellas tested positive. Vaccinated birds did not develop feather lesions, had only transient PCR detectable viraemia and had no evidence of persistent infection 270 days post-challenge using PCR, histopathology and immunohistochemistry (IHC). Non-vaccinated control corellas developed transient feather lesions and PCR, HI and HA test results consistent with PBFD. They were BFDV PCR positive for up to 41 days post-challenge and qRT PCR demonstrated reduced virus replication in vaccinated birds compared to non-vaccinated control birds. Thus, administration of recBFDVcap vaccine alone was found to incite an adaptive immune response in BFDV-free corellas that subsequently conferred protection against inoculation with BFDV. A commonly utilized method for excising blood dried on filter paper was proven to be of high risk of carryover contamination facilitated by a hole punch used for processing several samples. Therefore a practical method of avoiding carryover contamination was developed and used in the DNA testing procedures of the vaccination trial. Finally, the haematological characteristics of the above mentioned cockatoos were studied before and for 97 days after experimental infection with BFDV. It was found that the pre-challenge haematological values were similar between the vaccinated and non-vaccinated corellas. Most pre-challenge parameters were comparable to previously reported values of other cockatoos and psittacine birds. Significant differences were seen in both groups when comparing pre-challenge values with post challenge values for total and differential leukocyte concentrations, but PCV and TSP were not significantly affected by BFDV challenge.

Les animaux vivant au sein de groupe sociaux doivent gérer des interactions diverses et multiples avec leurs congénères. L’objectif de ma thèse portait sur les comportements sociaux chez les psittacidés et plus précisément l’évaluation de leur aptitude à adapter leurs comportements vis-à-vis d’un congénère ou d’un humain, qu’il s’agisse d’actions conjointes, d’attributions d’états mentaux ou de comportements prosociaux. Les oiseaux testés (perruche et gris du Gabon) ont été capables de coopérer et les perroquets ont appris à attendre le partenaire et à prendre en compte la nécessité de la présence d’un partenaire mais pas son rôle. Les quatre espèces de psittacidés testées n’ont pas profité de l’opportunité de récompenser un partenaire sans coût supplémentaire. Les gris du Gabon ont montré qu’ils étaient capables d’adapter leurs comportements en fonction de l’état attentionnel et des intentions d’un expérimentateur<br>Animals living in social groups have to manage divers and multiple interactions with their conspecifics. My thesis dealt with social behaviours in psittacids and more precisely the valuation of their ability to adapt their behaviours according to a partner whether it was cooperative actions, mental states attribution or prosocial behaviours. Tested birds (budgerigars and grey parrots) were able to cooperate and grey parrots learned to wait for the partner and took into account the necessity of the presence of a partner but not his role. The four psittacids species tested did not take the advantage to deliver food to a partner at no supplementary cost. Grey parrots showed that they were able to adapt their behaviours according to the attentional state and the intentions of an experimenter

Dans le règne animal, certaines espèces présentent une organisation en groupe complexe, permettant l’établissement d’interactions sociales plus ou moins élaborées entre les individus. Les comportements prosociaux, visant à améliorer le bien-être de l’autre, apparaissent préférentiellement entre animaux qui partagent une grande affinité ; ces comportements sont probablement favorisés par l’empathie, suggérant une prise en compte émotionnelle du partenaire. Les oiseaux, et particulièrement les corvidés et les psittacidés, se révèlent être d’excellents modèles pour étudier ce lien entre relation sociale et prise en compte de l’autre: ils forment des couples monogames sur le long terme au sein desquels une forte coopération existe. L’objectif de cette thèse est d’explorer comment le lien entre individus (familial, sexuel, amical) module les comportements prosociaux et empathiques. Les réactions des sujets sont évaluées via le partage de nourriture ou bien en observant une sensibilité accrue envers un congénère. Chez les perruches calopsittes (Nymphicus hollandicus), les oiseaux réagissent plus fortement au cri de détresse d’un oiseau préféré qu’au cri d’un autre congénère familier et les individus apparentés (frères et sœurs) partagent davantage la nourriture entre eux qu’avec les autres. Les perroquetsont tendance à être prosociaux mais la prise en compte de l’autre reste à confirmer. Les choucas des tours (Corvus monedula), placés dans une situation nouvelle avec un autre oiseau, passent plus de temps à proximité de leur partenaire sexuel que d’un autre oiseau de sexe opposé. Ces différents résultats suggèrent qu’un lien d’affinité existe entre les individus et qu’il façonne leurs comportements en termes de prosocialité et d’empathie<br>In the animal kingdom some species form complex social groups in which elaborated relationships between individuals occur. Prosocial behaviors, i.e. actions that benefit others, preferentially occur between closely affiliated individuals and may be driven by empathy, the ability to identify and share the emotional states of others. Birds, particularly corvids and parrots, are excellent candidates for investigating the link between social relationship and other-regarding behavior. They are long-lived and form long-term monogamous pair-bonds in which a high level of cooperation is seen throughout the year. The aim of this thesis is to study how the nature of a relationship (sibling, mate or friend) can modulate prosocial behavior and its underlying emotions in parrots and a corvid species. The approach was to study food-sharing or behavioral reactions to stressful situations such as distress call playback or exposure to novel objects, in different social contexts. Cockatiels (Nymphicus hollandicus) reacted more to the distress calls of a closely affiliated partner than to those of a non-partner, and they preferably shared food with affiliated, related individuals. Different species of parrots preferentially chose a prosocial option over a selfish one, but it remains unclear whether they took the other’s perspective into account. Confronted with intimidating novel objects, jackdaws (Corvus monedula) spent more time in mutual proximity when paired with their mate than when with a familiar opposite-sex non-partner. However, they were not bolder when accompanied by their mate compared to a non-partner. These results suggest that an emotional link exists between affiliated individuals and that this special bond drives their prosocial and empathic behavioral responses<br>Im Tierreich gibt es Arten mit komplexer Gruppenstruktur, in denen Individuen aufwendige soziale Beziehungen mit Artgenossen eingehen. Prosoziales Verhalten, ein Verhalten zum Wohle Anderer, tritt bevorzugt zwischen Individuen auf, die eine starke gegenseitige Bindung aufweisen. Prosozialität beruht auf Empathie-Fähigkeit, die wiederum ein gewisses Verständnis der emotionalen Lage von Artgenossen voraussetzt. Vögel, insbesondere Papageien und Krähen, sind geeignete Modelle, um Zusammenhänge zwischen sozialer Bindung und prosozialem Verhalten zu untersuchen: sie bilden Langzeit-monogame Paare, die das Jahr hindurch miteinander kooperieren. Ziel dieser Doktorarbeit ist es, herauszufinden, wie soziale Bindungen (verwandtschaftlicher, sexueller, oder freundschaftlicher Natur) prosoziales Verhalten und Empathie beeinflussen. Zu diesem Ziel wurden Studien über Futterteilen und Verhaltensreaktionen auf Stress in verschiedenen sozialen Kontexten durchgeführt. Nymphensittiche (Nymphicus hollandicus) reagierten stärker auf Warnrufe ihres Partners als auf die eines anderen Gruppenmitglieds. Ebenso teilten verwandte Sittiche häufiger Futter miteinander als mit anderen Individuen. Verschiedene Papageienarten bevorzugten in einer Entscheidungssituation prosoziale über egoistische Optionen, wobei unklar bleibt, ob sie die Perspektive ihres Partners verstanden. Dohlen (Corvus monedula), die mit einem für sie unheimlichen neuen Objekt konfrontiert wurden, verbrachten mehr Zeit in gegenseitiger Nähe, wenn sie mit Ihrem Partner als mit einem anderen Individuum getestet wurden. Sie verhielten sich aber in Gegenwart ihres Partners nicht mutiger. Die Ergebnisse legen nahe, dass Bindungen zwischen Individuen prosoziales Verhalten und empathische Reaktionen aufeinander beeinflussen

Internal papillomatous disease (IPD) is characterized by mucosal papillomas occurring primarily in the oral cavity and cloaca of Neotropical parrots. These lesions can cause considerable morbidity, and in some cases result in mortality. Efforts to demonstrate papillomavirus DNA or proteins in the lesions have been largely unsuccessful. However, increasing evidence suggests that mucosal papillomas may contain psittacid herpesviruses (PsHVs). In this study, PsHV 1 genotype 1, 2, and 3 DNA was found in 100% of mucosal papillomas from 30 Neotropical parrots by PCR using PsHV specific primers. However, Psittacus erithacus papillomavirus and finch papillomavirus DNA were not detected. Additionally, a novel PsHV sequence related to, but phylogenetically distinct from PsHV 1, was identified in 4 African grey parrots (Psittacus erithacus), two of which exhibited papillomas. These findings suggest that mucosal papillomas may develop in parrots latently infected with PsHV. Tumors of the bile and pancreatic ducts have also been observed in parrots with IPD. Other mucosal tumors including carcinomas of the proventriculus and ventriculus may be coincident with bile duct tumors, but cloacal carcinomas usually develop as solitary lesions. To test whether PsHV was associated with these tumors, the fresh tissues from 11 parrots and the formalin-fixed paraffin-embedded (FFPE) tissues of 5 parrots exhibiting mucosal tumors were examined by PCR. All tumors were found to contain PsHV 1 genotype 3 DNA except one bird with a cloacal carcinoma that contained genotype 4. Histologically normal tissues available from six parrots did not contain PsHV DNA. Experiments were performed using the FFPE tissues of 5 parrots with IPD related tumors known to contain PsHV by PCR, to show that the virus was in significantly higher concentration in the neoplastic tissue compared to adjacent histologically normal tissue. Neoplastic and adjacent unaffected cells were dissected from the tissues using laser capture microdissection and the DNA was examined by PCR. In situ hybridization using PsHV specific probes and direct in situ PCR were also performed on the tissues. A strong association was shown between infection by PsHV 1 genotype 3 and birds manifesting IPD related tumors and other neoplasms of the digestive tract.

The circovirus Beak and feather disease virus (BFDV) causes psittacine beak and feather disease (PBFD) that is characterised by a chronic disease process associated with feather abnormalities, beak deformities and eventual death in various species of birds in the order Psittaciformes. This disease is seen in captive and wild psittacine species in Australia and several other countries and is a significant threat to the survival of some endangered psittacine species. This thesis reports on genetic studies that have furthered the understanding of the diversity of BFDV present within Australia. These studies have optimised methods of detecting BFDV. They have also resulted in the production of an immunogenic and antigenic recombinant BFDV Capsid protein that could lead to alternate methods of producing viral antigen for serological tests and the development of a BFDV vaccine. To assess the optimal method of the detection of BFDV infection, feather and blood samples were submitted by referring veterinarians throughout Australia from psittacine birds tentatively diagnosed with PBFD or with a history of being in contact with PBFD-affected birds. These samples were examined by 3 procedures commonly used to detect BFDV infection: a polymerase chain reaction (PCR) assay and haemagglutination (HA) for the detection of virus, and haemagglutination inhibition (HI) tests for the detection of virus antibody in response to infection. Of the samples examined from 623 psittacine birds, the prevalence of BFDV DNA in feather samples detected by PCR was 18.85%. There was a strong correlation between PCR and HA testing of feather samples, although possible false-positive and false-negative PCR and HA results were obtained in some samples. Of the 143 birds that were PCR feather-positive only 2 had detectable HI antibody and these birds were also HA feather-negative, which suggests that they were developing immunity to recent infection. All birds with HI antibody were feather HA negative. Despite the rare occurrence of PBFD in cockatiels (Nymphicus hollandicus), 2 of the 13 samples collected from this species were PCR and HA positive indicating that this species can be infected with BFDV. Three studies were undertaken to further our understanding of the genetics of BFDV in Australian avifauna: sequence analysis of the BFDV detected in a grey cockatiel (Nymphicus hollandicus), a species normally considered resistant to infection with BFDV; analysis of the genome of BFDV present in lorikeets (Trichoglossus sp.) in Australia; and analysis of the genome of BFDV detected in endangered swift parrots (Lathamus discolor). Sequence analysis of the entire genome of the cockatiel BFDV isolate revealed that it clustered phylogenetically with 2 other viruses, one from a sulphur crested cockatoo (SCC1-AUS) and one from a Major Mitchell cockatoo (MMC-AUS), which suggests that this isolate from the grey cockatiel was not a cockatiel-specific biotype. Phylogenetic analysis of the ORF V1 of BFDV detected in 7 lorikeets demonstrated these 7 isolates clustered phylogenetically with other BFDV isolates obtained from Loriidae species elsewhere in the world and confirmed the presence of a loriid-specific genotype. Phylogenetic analysis of the sequence data generated from ORF V1 of virus detected in 2 endangered swift parrots provided evidence they were also infected with BFDV genotypes derived from other species of birds, one isolate clustering with viruses from a Loriidae genotype and the other with isolates derived from species of Cacatuidae and Psittacidae. As part of this research, a baculovirus expression system was successfully developed for the production of recombinant BFDV Capsid protein. Inoculation of this protein into chickens resulted in the development of HI antibody, which demonstrated its immunogenicity. When used as an antigen in HI tests it detected antibody in virus-infected birds, which demonstrated its antigenicity. This protein offers potential application as an antigen for the development of serological tests and as an immunogen for incorporation into vaccines for control of PBFD.

Murdoch University (Ph.D.)--Murdoch University, 2008.<br>Contains three published journal articles at back of thesis. Thesis submitted to the Faculty of Health Sciences. Includes bibliographical references (leaves 196-231)

Beak and Feather Disease Virus (BFDV) is a circovirus which causes ill-thrift, feather loss and immunosuppression leading to secondary infections and eventually death in psittacine birds. The development of standardised reagents for the detection and characterisation of BFDV infections and for the production of protective vaccines has been difficult as no cell culture system has yet been found to grow the virus successfully in vitro. However, the development of consistent and effective diagnostic tests and vaccines is now more practical through the application of nucleic acid-based detection methods and recombinant technology. A quantitative real-time PCR assay for the detection of BFDV DNA was developed, using primers designed to amplify a conserved 81 bp fragment of ORFV1 and SYTO9, a fluorescent intercalating dye, with assays run on a Corbett RotorGene 3000. A synthetic oligonucleotide was used to establish standard curves for the quantitation of viral load in both blood and feather preparations. The assay was very sensitive, with a detection limit of 50 copies/ìL. The assay was developed using BFDV-positive DNA extracts from the feathers of 10 different species of birds and validated with blood and feather samples from corellas vaccinated with an experimental BFDV vaccine, then challenged with live virus. Viral DNA was reliably detected in the blood of all control (non-vaccinated) birds and in some vaccinated birds. Contamination of the environment with the feather dander of BFDV-infected birds meant that HA feather preparations were unreliable for the detection and quantitation of viral excretion. Nonetheless, the assay should prove to be a useful and sensitive test for the detection of viral DNA in a range of samples in future investigations. A recombinant BFDV capsid protein was also produced and a specific monoclonal antibody developed against it. The behaviour of the protein in haemagglutination (HA) assays and the behaviour of the monoclonal antibody in western blotting, immunohistochemistry (IHC), ELISA and haemagglutination-inhibition (HI) assays were characterised. The protein had the ability to agglutinate galah erythrocytes as per the wild-type virus and this agglutination was successfully inhibited by antibodies to wild-type BFDV from naturally immune psittacine birds. Furthermore, the protein self-assembled into virus-like particles as determined by electron microscopy. The antibody was specific for both the recombinant BFDV capsid protein and the whole virus and had similar optimal titres when used in western blotting and IHC. The antibody also had HI activity and detected BFDV virus from 3 genera of psittacine birds, including the recently described cockatiel BFDV isolate. A novel “blocking” (or “competitive”) ELISA (bELISA) for the detection of anti- BFDV antibodies in psittacine sera (Ab-bELISA) was also developed and validated with 166 samples from eastern long-billed corellas vaccinated with the recombinant capsid protein and challenged with live virus. The bELISA was found to be both sensitive and specific and correlated strongly with the HI test, thus it should have wide application for the serodiagnosis of BFDV. A survey of cockatiels (n=88) housed at commercial aviaries was conducted to investigate whether BFDV infection occurs in cockatiels. All birds were diagnosed as being virus-free by PCR and HA and had no detectable antibody titre by HI assay. In addition to this, the genomes of two BFDV isolates obtained from diseased cockatiel feathers were sequenced and cross-reactivity assays performed using virus eluted from these feathers and sera from naturally immune psittacine birds. Serological cross-reactivity results and phylogenetic analysis of the nucleotide sequences indicated that the cockatiel virus isolates were serologically and genetically different to other BFDV isolates. This is the first report of an antigenically distinct BFDV in psittacine birds. Since the Ab-bELISA has a lower limit of detection than the HI assay, it was used to repeat the cockatiel sero-survey. No antibodies were detectable in any of the cockatiels tested and thus questions about the real prevalence of BFDV infection in cockatiels and the possible existence of a novel BFDV serotype adapted to cockatiels remain unanswered. The successful control of PBFD in both pet and wild birds depends on the development of vaccines that incite a strong specific immune response and can be efficiently produced in large quantities. Recombinant BFDV capsid proteins have recently been considered as candidate vaccines against BFDV and recombinant techniques allow the development of other candidate vaccines, including DNA vaccines. In order to examine the potential of DNA vaccination as a strategy for the prevention and control of BFDV, two DNA vaccines, based on the nucleotide sequence encoding the capsid protein of BFDV, were developed using the mammalian expression vector pVAX1. The vaccine constructs encoding both the full length and NLS-truncated capsid protein resulted in protein expression both in vitro and in vivo. Protein was detected in COS-7 cells transfected with the constructs with an indirect immunocytochemistry assay using the monoclonal antibody described in Chapter 5. Protein was present in the nucleus of cells transfected with the vaccine encoding the full-length nucleotide sequence and in the cytoplasm of cells transfected with the vaccine encoding the NLS-truncated sequence as expected. Both DNA vaccine constructs induced detectable levels of anti-BFDV antibodies in vaccinated birds, determined using the Ab-bELISA described in Chapter 5. Thus, DNA vaccines similar to those presented here may have application in the prevention and control of BFDV and some options for the further development of these vaccines into effective methods for the control of BFDV are discussed.

Dissertação de Mestrado Integrado em Medicina Veterinária<br>A Doença de Dilatação do Proventrículo (DDP) foi inicialmente descrita na década de 1970 nos Estados Unidos da América (EUA) e na Europa. Trata-se de uma doença fatal, que atinge mais de 50 espécies diferentes de aves domésticas e silvestres por todo o mundo. Esta doença é caracterizada por infiltrações linfo-plasmocíticas dos gânglios do sistema nervoso central e periférico, originando alterações neurológicas e/ou disfunções do aparelho digestivo. A epidemiologia desta doença, associada ao facto de um vírus desconhecido com envelope ter sido frequentemente observado, em tecidos e/ou fezes de animais doentes, sugere uma etiologia viral. Recentemente o Bornavirus Aviário, um novo género da família Bornaviridae, foi relacionado com psitacídeos afectados com DDP. Neste estudo, relatamos doze casos observados na Catalunha, Espanha. Descrevemos os sinais clínicos, os resultados de exames complementares de diagnóstico, as necrópsias e os exames histopatológicos realizados. Um dos obstáculos com que nos deparámos foi a dificuldade de diagnóstico em vida e pós-mortem, apenas um dos doze animais foi diagnosticado com DDP antes de morrer e quatro após a necrópsia. No entanto os restantes sete casos apresentavam sintomatologia compatível assim como relação temporal e geográfica com as aves positivas. Assim, pensamos que será importante, na actualidade, desenvolver métodos menos invasivos de diagnóstico baseados na biologia molecular para a detecção de Bornavirus Aviário e de testes serológicos como alternativa aos métodos histopatológicos. No futuro deverão ser desenvolvidos em Portugal estudos epidemiológicos sobre DDP, envolvendo os médicos veterinários, proprietários, a comunidade científica e entidades oficiais com objectivo de avaliar os possíveis impactos económicos na avicultura e na protecção da avifauna Portuguesa.<br>ABSTRACT - PROVENTRICULAR DILATATION DISEASE: A BORNAVIROSIS - Proventricular dilatation disease (PDD) was first described in the 1970’s in the USA and was also reported in Europe. Is a fatal disorder identified in over 50 different domesticated and wild birds species worldwide. The disease is characterized by lymphoplasmocytic infiltration of the ganglia of the central and peripheral nervous system, leading to central nervous system disorders and/or enteric motility disfunction. The epidemiology of the disease, along with the fact that an unknown virus with an envelope was frequently observed in affected tissues and/or feces of diseased birds, suggests a viral etiology. Recently Avian Bornavirus, representing a new genus within the family Bornaviridae, was related with PDD positive psittacine birds. This study reports twelve cases seen in Catalunya, Spain. Clinical signs, complementary diagnosis exams results, necropsy and histopathological findings are described. We found it very dificult to achive the final diagnosis in alive birds and at post-mortem, only one animal was diagnosed with PDD before dying and four after necropsy. However, the remaining seven cases showed compatible signs as well as temporal and geografic relation with the positive birds. Therefore it is important to develop new diagnostic methods based on molecular biological detection of Bornavirus, as well as serological tests as viable alternatives to invasive and postmortem histopathological diagnosis. Future studies on the epidemiology of PDD must be implemented in Portugal, combining efforts of private veterinarians, owners, scientifical community and Government authorities to evaluate the possible devastating impact on aviculture and on the Portuguese wild avifauna.

Dissertação de Mestrado Integrado em Medicina Veterinária<br>A dissertação aqui presente estuda pela primeira vez, a infecção pelo circovirus aviário (Beak and Feather Disease Virus - BFDV) em aves psitacídeas na região espanhola da Catalunha, entre 2005 e 2008. O BFDV provoca a doença do bico e penas dos psitacídeos (Psittacine Beak and Feather Disease - PBFD), cuja apresentação clínica depende de vários factores relativos à ave infectada, ao ambiente e ao genótipo viral. A doença é observada em aves de cativeiro de todo o mundo e em aves selvagens de África e do Pacífico, colocando em risco a conservação de algumas espécies ameaçadas. É apresentada uma revisão bibliográfica sobre o tema abordando a caracterização da doença em termos de etiologia, patogenicidade, quadros clínicos, diagnóstico, tratamento e profilaxia. Através de amostras maioritariamente sanguíneas de 1348 animais de pelo menos 83 espécies diferentes, foi testada pelo método da reacção em cadeia da polimerase (PCR) a presença do DNA viral. Os animais estudados são na esmagadora maioria nascidos em cativeiro em países europeus e residentes da região. Tendo em conta os resultados de estudos anteriores e de diferentes regiões geográficas, encontrou-se um valor de prevalência relativamente reduzido (2,60%±0,8%). Certos grupos taxonómicos revelaram maior prevalência da infecção (Ecletus sp., Agapornis sp. Ara sp. e Psittacus erithacus ssp.), enquanto noutros não se encontraram animais positivos em todo o estudo. Não foi encontrada qualquer associação estatística entre a infecção por BFDV e o sexo ou a presença de Chlamydophila sp. Foi estudado o quadro clínico de 26 animais positivos e foi possível observar que 49% não apresentavam sintomas no momento da colheita de sangue. O quadro clínico agudo apenas se observou em Psittacus erithacus ssp. e Ecletus roratus ssp.<br>ABSTRACT - Studies on the detection of Avian Circovirus in domestic psittacines in Barcelona region - Spain - ~This dissertation studies for the first time the avian circovirus (BFDV) infection in psittacine birds in the Spanish region of Catalonia, between 2005 and 2008. BFDV is the etiologic agent of the Psittacine Beak and Feather Disease (PBFD), whose clinical presentation depends on various factors relative to the infected bird, environment and viral genotype. The disease is observed in captive birds worldwide and in wild birds in Africa and Pacific, putting at risk the conservation of some endangered species. A literature review is presented approaching the etiology, pathogenicity, clinical presentation, diagnosis, treatment, and prophylaxis. The presence of viral DNA was tested by PCR in blood samples from 1348 animals of about 83 different species. Most of the tested animals were captive born in European countries and living in Catalonia region. Comparing to other similar studies and from different geographic regions, the prevalence value was relatively low (2,60%±0,8). Certain taxonomic groups revealed higher infection prevalence (Ecletus sp., Agapornis sp., Ara sp. e Psittacus erithacus ssp.), while in other species no positive birds were found in the whole study. No statistical relation was found between BFDV infection and sex or the presence of Chlamydophila sp. The clinical presentation in 26 positive animals was studied and it was observed that 49% didn’t show any symptoms at the moment of the blood sample collection. The acute clinical presentation was observed in Psittacus erithacus ssp. and Ecletus roratus ssp. species.

Um componente da conservação de animais selvagens é a relocação de espécies comumente referida como projetos de soltura. Para que esta seja bem sucedida os candidatos do projeto devem estar livres de patógenos considerados de importância para a espécie. Segundo a Instrução Normativa 179 oficializada pelo IBAMA em 25/06/2008, determinou-se a realização de exames laboratoriais como medidas a se prevenir a introdução de agentes infecciosos no ambiente, e garantir a sobrevivência a longo prazo dos animais em questão. No Brasil encontra-se a maior quantidade em espécies de psitacídeos, e das 84 espécies, 13 são vulneráveis a criticamente ameaçadas de extinção. Diversos projetos de relocação de animais silvestres, incluindo vários já bem sucedidos com psitacídeos, vêm sido realizados em território nacional além dos existentes do exterior. O Papagaio-de-peito-roxo (Amazona vinacea) tem suas populações severamente afetadas, sendo classificada no estado de São Paulo como criticamente ameaçada e como ameaçada a nível mundial. Apesar das dificuldades para a conservação dos recursos naturais, existem remanescentes de habitat protegido e em regeneração que podem abrigar espécies que historicamente ocupavam estes locais, de maneira que projetos de reintrodução visando A.vinacea como espécie bandeira em São Paulo e em Santa Catarina, foram contatados para realizar a pesquisa sanitária prévia à soltura. Foram realizados suabes cloacais em amostragens seriadas de modo a identificar possíveis portadores para os agentes paramixovírus tipo 1, influenza tipo A, poxvírus aviário, coronavírus, Escherichia coli Enteropatogênica (EPEC), e Salmonella spp., em indivíduos da espécie confiscados do tráfico, através da reação em cadeia pela Polimerase (PCR), objetivando sequenciar os possíveis resultados positivos, discutindo a viabilidade e custos segundo as determinações da IN 179/2008. Amostras após a liberação também foram coletadas na forma de fezes obtidas no recinto de aclimatação e/ou ao redor dos comedouros de alimentação suplementar. Obtiveram-se um total de 151 amostras somadas em todas as coletas seriadas, sendo 103 amostras de suabes cloacais de aves ainda em cativeiro e 48 amostras de fezes não individualmente caracterizadas das aves soltas. Das amostras testadas para os agentes com potencial infeccioso obtiveram-se apenas resultados positivos para E.coli, totalizando 36 isolados, embora nenhum tenha sido caracterizado como EPEC. Observou-se uma tendência para maior detecção de E.coli nas amostragens iniciais em comparação com as finais, fato ligado principalmente às melhorias no manejo empregadas. A possibilidade de se comparar resultados em amostragens seriadas foi importante para uma avaliação mais segura quanto à sanidade dos animais envolvidos, auxiliando a determinar a seleção das aves, não havendo relatos de doenças imediatamente após a soltura ou no monitoramento a longo prazo. A baixa frequência de amostras positivas para os agentes que poderiam inviabilizar a liberação parece demonstrar que existe um exagero de que doenças representam um risco extremo impedindo projetos de relocação. Procedimentos de quarentena adequados e a realização de um mínimo de exames reduzem os riscos envolvidos, fato observado no presente estudo tanto em cativeiro como no processo de liberação e posteriormente. Para as instituições amostradas havia um limitado recurso anual que não seria capaz de pagar por exames individuais. Uma opção é a de testar amostras em pool para diminuir os custos, e caso haja positivos, procurar retestar para identificar os indivíduos. A baixa prevalência de positividade para os agentes com potencial infeccioso neste estudo, demonstra que amostras em pool podem ser uma alternativa econômica, caso o exame individual esteja fora de questão. Tendo em vista que para obter um mínimo de segurança no projeto de relocação no Brasil depende-se quase exclusivamente da iniciativa privada, convênios com universidades tornam-se não apenas uma necessidade, mas também uma oportunidade para troca em direção a um objetivo em comum e geração de conhecimento científico.<br>One component in the conservation of wild animals is the relocation of species, commonly referred as release projects. In order for this attempt to be successful the candidates must be clear of pathogens of significance for the species. According to the normative rule 179 established by the IBAMA in 25/06/2008, it was determined that a series of laboratorial exams should be performed in order to prevent the introduction of infectious agents in the environment and guarantee the long term survival of the animals. The largest number of psittacine species is found in Brazil accounting for 84 species, with 13 of these classified as vulnerable to critically endangered. Several relocation projects with wild animals, including several well succeeded with psittacines have been taking place on a national scale besides others being carried out around the world. The Vinaceous Amazon (Amazona vinacea) have had its populations severely affected being classified as critically endangered in the state of São Paulo and globally threatened. Although there are challenges to conserve natural resources, available remnants of protected and regenerating habitat can be found and could support species that historically inhabited these sites, hence reintroduction projects with A.vinacea as a flagship species in the state of São Paulo and Santa Catarina were contacted to perform a health survey previously to the releases. Cloacal swabs were taken in paired samplings in order to detect possible carriers for paramyxovirus typ1, influenza type A, avian poxvirus, coronavirus, Enteropathogenic Escherichia coli (EPEC), and Salmonella spp.; in individuals that were confiscated from the illegal trade employing the Polymerase Chain Reaction (PCR), aiming to sequence possible positive results and discussing the viability and costs according to the determination of the IN/179/2008. Fecal samples were also collected after the release in the acclimation flight and/or surrounding the supplemental feeders in the area while still adapting in the post-release. A total of 151 samples were obtained altogether with 103 of these being cloacal swabs of the birds still in captivity and 48 fecal samples non individually characterized of released birds. Out of the tested samples only E.coli yielded positive results with a total of 36 isolates, although none was characterized as EPEC. A tendency was observed for a higher detection of E.coli in the initial samplings compared to the final ones, a fact mainly connected husbandry improvements that were put in use. The possibility to compare results in paired samplings was important in order to obtain a safer evaluation concerning the health status of the animals, helping to determine the birds selection and no health problems reported neither immediately after the release nor on the long term monitoring. The low frequency of positive samples for the agents that could jeopardize a release seems to show that there is an exaggeration that diseases represent an extreme risk to the point of hampering relocation projects. Adequate quarantine procedures and performing minimum health examinations minimize the involved risks, a fact observed in this study both in captivity as well as in the release and post release process. For the studied institutions there was a limited annual budget that would not be capable to pay for individual exams. One option is to test pooled samples to lower associated costs, and if a positive is found, retest to identify the individuals. The low prevalence for the tested agents in this study show that pooled samples could be a viable alternative when individual exams are not feasible. Taking into account that to obtain a minimum in terms of safety for a relocation project in Brazil one is almost exclusively dependent on private parties, cooperation with universities are not only a necessity but also an opportunity for exchanges toward a common goal besides generating scientific knowledge.

Orientação : Maria Margarida Ferreira Alves ; co-orientação : Filipe Miguel Reis Martinho<br>Measuring newborns weight gain per day can be a useful tool to determine if these baby chicks are growing as expected for a healthy bird. Although its importance is acknowledged, current information on what is the expected weight gain/day of a newborn psittacine is limited. The general goal of the present thesis is to determine the standard value of weight gain/day in selected species of psittacines. This study aimed to provide charts and tables and supply a reliable framework for those interested in pediatric psittacine health. All data was obtained in Loro Parque from chicks incubator-born or nestborn from January 2011 to December 2011. For the descriptive analysis SPSS/PASW 20 IBM was used. Disease was taken into account, so animals were stratified into healthy and sick groups. Weight mean in grams by day and respective upper and lower confidence interval (CI) were found. A CI of 95% was applied. All growth curves were obtained with these data. When N<3 (N being the number of psittacines of one species) both upper and lower CI withdraw from the media line, decreasing the confidence of the results. When 3<N<8 an increased confidence of the results was observed. When N>8 both upper and lower CI stood close to the media line, increasing the confidence of the results. These results are in conformity with what was expected. Curves from the following species reached this study’s purpose, being representative of their growth: Ara glaucogularis, Aratinga solstitialis, Cacatua galerita, Poicephalus robustus, Psittacus erithacus, Pyrrhura perlata and Trichoglossus haematodus. This is the first study on weight gain/day for these species. With this study it was possible to elaborate and provide tables with the expected weight/day of a healthy psittacine chick from hatching until fledging.

碩士<br>國立臺灣大學<br>獸醫學研究所<br>92<br>Avian polyomavirus (APV) and psittacine beak and feather disease virus (PBFDV) are the common viral disease of psittacine birds. APV belongs to genes Avipolyomavirus of the family Papovaviridae, and causes acute fatal disease in young budgerigar with a mortality rate of up to 100%. APV infection is also known as budgerigar fledgling disease (BFD). PBFDV belongs to genes Circovirus of the family Circoviridae. PBFDV affects over 60 species of wild and captive psittacine birds. Clinical signs include chronic and progressive losses of feathers and in some species deformities of the beak and claws. In this study, polymerase chain reaction (PCR) was used to detect nucleic acids of APV and PBFDV for epidemical investigation. The results showed an APV positive rate of 28.8%, PBFDV positive rate of 55.1%, and mixed infections positive rate of 24.6%. By cloning and sequencing, sequences of the PCR products were compared with the sequence obtained from the GeneBank. In PBFDV sequences, the identity of VP1 genome and T antigen coding region genome sequences for Taiwan ranges 94.6-97.3% and 90.1-99.1%, respectively. In PBFDV sequences, the identity of ORF1 and C1 sequences ranges 87.5-97.3% and 86.5-98.4%, respectively. Base on nucleotide and amino acid sequencing analyses, there were variation in the VP1 genome and T antigen coding region genome sequences from that of the APV and samples from the GeneBank. Derived amino acid sequence alignment for PBFDV ORF1 fragments, demonstrates the replication motifs (YCKS) and the nucleotide binding site motif (GPPGCGKS). In PBFDV C1 amino acid sequence have four positions considered as hypervariable.

Traineeship Report<br>The internship referred to in this report was carried out at the Avian and Exotic Clinic of the University of Veterinary and Pharmaceutical Sciences, in Brno, Czech Republic. This summarises a three-month work, between 17th September and 17th December 2018. The first chapter provides the state-of-the art regarding the Psittacine Beak and Feather Disease (PBFD), which gathers a review of the literature. The second chapter presents a practical study about PBDF, in which the Clinic was involved and that I had the opportunity to follow. The third chapter summarizes the casuistry observed during the internship. PBFD (Psittacine Beak and Feather Disease) is one of the most important worldwide viral diseases in psittacines and is caused by a circovirus. This virus is present in both wild and captive birds and can affect all ages, but juveniles and young adults are particularly susceptible. PBFD has different courses, depending on the bird’s age at the time of the infection and the presence of secondary infections, which may reflect in a peracute, acute or chronic form. However, the existence of subclinical infections is a major issue regarding the virus detection and spread. For this reason and because this virus is highly contagious among psittacine species, there is an increasing necessity to combine preventive care measures and early diagnostics in clinical practice.<br>O estágio curricular apresentado neste relatório teve lugar na Clínica de Aves e Animais Exóticos da Universidade de Ciências Veterinárias e Farmacêuticas, em Brno, República Checa. Este é um resumo de três meses de trabalho, entre 17 de Setembro e 17 de Dezembro de 2018. O primeiro capítulo consiste numa monografia relativa à doença do bico e das penas dos psitacídeos (Psittacine Beak and Feather Disease - PBFD), e reúne a revisão bibliográfica acerca do tema. O segundo capítulo apresenta um estudo prático acerca da PBFD, no qual a Clínica esteve envolvida e que tive a oportunidade de acompanhar. O terceiro capítulo sumariza a casuística observada durante o estágio. PBFD é uma das mais importantes doenças virais em psitacídeos de todo o mundo e é causada por um circovirus. Este vírus está presente tanto em aves selvagens como em cativeiro e pode afetar qualquer idade, contudo, neonatos e jovens são particularmente suscetíveis. PBFD tem diferentes apresentações dependendo da idade da ave na altura da infeção e da ocorrência de infeções secundárias, o que se pode refletir num quadro hiperagudo, agudo ou crónico. No entanto, a existência de infeções subclínicas é uma das principais preocupações no que diz respeito à deteção e disseminação do vírus. Por esta razão e porque este vírus é altamente contagioso entre espécies de psitacídeos, há uma necessidade crescente de aliar medidas de prevenção e deteção precoce na prática clínica.

The Psittacidae is one of the most endangered families of birds in the world. Knowledge of its nutrition is important for understanding their survival and productivity in the wild, as well as for their adequate husbandry in captivity. Hand-rearing is a common practice for this group. However, research on their requirements is limited. Analysis of the crop content of chicks can provide new insights into psittacine nutrition, but it is limited by the small sizes of samples which can be obtained. We sampled the crops from free-living chicks of scarlet macaws and red-and-green macaws from southeastern Peru, Cuban parrots from the Bahamas, lilac-crowned parrots from northwestern Mexico, and thick-billed parrots from northern Mexico. The predicted metabolizable energy, protein, fat, minerals, profile of essential amino acids and profile of fatty acids of the crop samples, as well as from 15 commercial hand-rearing formulas, were analyzed and contrasted. Near Infrared Spectroscopy was shown to be a valid technique for the nondestructive, low cost prediction of a variety of nutritional attributes of crop samples as small as 0.5 g dry weight, expanding the possibilities of wild animal nutrition research. The diets of the five studied species presented remarkable similarities and common patterns. The predicted dietary metabolizable energy and fat concentrations were particularly similar among species, the thick-billed parrot being the one with the most unique nutrient profile. The fatty acid profile of the crop contents differed markedly among genera, with the thick-billed parrot closer to the macaws than to the parrots. In comparison with the crop samples, the hand feeding formulas presented lower fat, Mg, arginine, and valine concentrations. The wide variation in nutrients suggests that there is not yet a consensus among manufacturers concerning the correct nutrition for growing psittacines. It is suggested that a single formulation could be used to hand-rear macaws and parrots from half its nesting time to fledging, and further research should focus on their nutrition during the first half. Our results suggest that manufacturers should evaluate if increasing the concentrations of crude fat, Mg, arginine, and valine in commercial formulas enhances psittacine chick growth and health.

碩士<br>國立屏東科技大學<br>獸醫學系所<br>106<br>Polyomavirus (APV) mainly causes acute systemic symptoms, also known as small parrot disease, which happens on the psittacine chicks. All age of the parrot birds could potentially been infected. If being infected, the death rate of clinical symptoms is related to the age of infection. Part of the chicks may die suddenly before the clinical symptoms of acute appeared; whereas the chicks that survive will have the appearance of clinical symptoms. These symptoms of the survival include obvious ones: symmetrical drop-out of feathers, abdominal distention, dyschromia, subcutaneous hemorrhage, and sometimes neurological symptoms the tremor of head and neck ataxia. Psittacine beak and feather disease virus (PBFDV) can infect parrot. The clinical symptoms include weight loss, underdeveloped feathers, hair removal and beak malformation. PBFDV also damages on the immune tissues and organs, suppress the immune system, and even cause pneumonia and sepsis enteritis. If severe it causes death. PBFD has very high incidence. The study will take samples from the Northern, Central and Southern parts. Sixty samples will be collected in every month, and a total of 720 blood samples will be collected in a year. Each sample were extracted DNA, then use the primers to polymerase chain reaction (PCR) to in the document. The result is complete PCR assay for detection technology, preliminary results showed that APV and PBFDV ratio both in the spring and fall with the highest occurrence, whereas most proportion occur in psittacine chicks, Large farms and Psittacula krameri. PBFD in psittacus is most proportion.

碩士<br>中興大學<br>獸醫學系暨研究所<br>99<br>Psittacine beak and feather disease (PBFD) is a progressive symmetric feather dystrophy, which is caused by a circular, single-strand DNA virus belonging to Circoviridae Circovirus. In Taiwan, one of the previous studies based on clinical cases revealed that positive rate was 41.2% (68/165) during 2002-2005 (Hsu et al., 2006), and another one based on clinical cases, bird parks and independent breeding facilities during 2007-2008 that positive rate was 31.8%(408/1285)( Kuan et al., 2009). Since that Taiwan is not one of the countries of origin for parrots, the imported parrots should be an important pathway for the transmission of PBFDV. In this study, we collected the samples from imported parrots during JUN to SEP in 2010 in Taiwan, and tested the samples with the PCR to monitor the PBFDV. As the results, the positive rate was 59.96% (45/79) in JUN, 90.48% (57/63) in JUL and 72.58% (45/62) in SEP; the total positive rate was 72.06% (148/204). On the basic of the source (new world and old world), the positive rate were 82.56% (71/86) in new world genera and 64.41% (76/118) in old world genera. The odds ratio (OR) was 2.62 between the two sources, it showed that different sources was not significantly associated with the positive rate in imported parrots. The OR between imported parrots and captive parrots in Taiwan was 5.29, showed that the imported parrots is really an important pathway for the transmission of PBFDV. In our study, we separated a complete sequence of PBFDV (Poi-TW03_HYT) and four ORF C1 sequences of PBFDV (Tan-TW02_HYT, Poi-TW03_HYT, Tan-TW04_HYT and Poi-TW05_HYT); alignment of the isolates with the sequences from GeneBank and the previous studies in Taiwan.Tan-TW02_HYT and Tan-TW04_HYT show the species differences, but Poi-TW03_HYT and Poi-TW05_HYT were clustered with the different species. The established phylogenetic tree did not support the existence of specific genotypes in Taiwan.

碩士<br>國立臺灣大學<br>獸醫學研究所<br>94<br>Avian polyomavirus (APV) infection and psittacine beak and feather disease (PBFD) are the most common viral diseases of psittacine birds. APV was first isolated from budgerigars in the early 1980s. APV belongs to genes Avipolyomavirus of the family Papovaviridae and causes acute fatal disease in young budgerigars with 100% mortality rate. APV infection is also known as budgerigar fledgling disease. PBFDV, categorized into genes Circovirus of the family Circoviridae, affects over 60 species of wild and captive psittacine birds. In this study polymerase chain reaction (PCR) was applied to diagnosis either APV or PBFDV infections in psittacine birds of Taiwan. From 2002 to 2005, the positive rate of APV, PBFDV, and APV/PBFDV infection over 165 cases were 15.2%, 41.2%, and 10.3% respectively. APVs indicated over 97% nucleotide identity in VP1 and T antigen coding regions. PBFDVs had over 92.2% and 83.3% nucleotide identity in ORF V1 and ORF C1 sequences. Another aim of this study is to prepare Sandwich ELISA for detection of APV and to analyze the specificity and sensitivity of Sandwich ELISA. First of all, we applied prokaryotic (E. coli.) system to express the viral structural protein VP1. The expressed VP1 were specifically recognized by his-tag and anti-APV antibodies. The recombinant protein was used as an immunogen to inject BALB/c mice for preparation of hybridomas which produced anti-VP1 antibodies. Eight monoclonal antibodies secreting hybridomas were obtained as determined by ELISA and Western blot. After screening of these 8 monoclonal antibodies, one monoclonal antibody, D10-6, is used to develop the Sandwich ELISA for the detection of APV. No significant difference was formed between PCR and Sandwich ELISA at the sensitivity. The Sandwich ELISA could be applied for diagnosis of APV infected budgerigars, and could further be applied to immunoassay strip for the rapid diagnosis of APV infection in budgerigars.