Avipoxvirus detected in tumor-like lesions in a white-faced whistling duck (Dendrocygna viduata) / Avipoxvirus detectado em lesões tipo tumorais em uma marreca-irerê (Dendrocygna viduata)

Avipoxvirus is the etiological agent of the avian pox, a well-known disease of captive and wild birds, and it has been associated with tumor-like lesions in some avian species. A white-faced whistling duck (Dendrocygna viduata) raised in captivity was referred to a Veterinary Teaching Hospital in Northeast due to cutaneous nodules present in both wings. A few days after the clinical examination, the animal died naturally. Once submitted to necropsy, histopathological evaluation of the lesions revealed clusters of proliferating epithelial cells expanding toward the dermis. Some of these cells had round, well-defined, intracytoplasmic eosinophilic material suggestive of poxvirus inclusion (Bollinger bodies). PCR performed on the DNA extracted from tissue samples amplified a fragment of the 4b core protein gene (fpv167), which was purified and sequenced. This fragment of Avipoxvirus DNA present in these tumor-like lesions showed high genetic homology (100.0%) with other poxviruses detected in different avian species in several countries, but none of them were related to tumor-like lesions or squamous cell carcinoma. This is the first report of Avipoxvirus detected in tumor-like lesions of a white-faced whistling duck with phylogenetic analysis of the virus.(AU)

Avipoxvirus é o agente etiológico da varíola (bouba) aviária, uma doença bem descrita em aves de cativeiro e selvagens, tendo sido associada a lesões semelhantes a tumores em algumas dessas espécies. Uma marreca piadeira (Dendrocygna viduata), criada em cativeiro, foi atendida em um Hospital Veterinário na região nordeste devido à presença de nódulos cutâneos em ambas as asas. Alguns dias após o exame clínico, o animal veio a óbito naturalmente. A ave foi submetida à necropsia e coletados fragmentos das lesões para análise histopatológica, que revelou proliferação de células epiteliais expandindo para a derme. Algumas dessas células possuíam material eosinofílico intracitoplasmático e bem definido, sugestivo de inclusão de poxvírus (corpúsculos de Bollinger). A PCR realizada a partir do DNA extraído de amostras das lesões amplificou um fragmento do gene da proteína do núcleo 4b (fpv 167), que foi purificado e sequenciado. Esse fragmento de DNA de Avipoxvirus presente nas lesões relevou alta homologia genética (100,0%) com outros poxvírus detectados em diferentes espécies de aves em vários países, mas nenhum deles estava relacionado a lesões tumorais ou carcinoma espinocelular. Este é o primeiro relato de Avipoxvirus detectado em lesões semelhantes a tumores em uma marreca piadeira com caracterização molecular do vírus.(AU)

Avipoxvirus detected in tumor-like lesions in a white-faced whistling duck (Dendrocygna viduata) / Avipoxvirus detectado em lesões tipo tumorais em uma marreca-irerê (Dendrocygna viduata)

Avipoxvirus is the etiological agent of the avian pox, a well-known disease of captive and wild birds, and it has been associated with tumor-like lesions in some avian species. A white-faced whistling duck (Dendrocygna viduata) raised in captivity was referred to a Veterinary Teaching Hospital in Northeast due to cutaneous nodules present in both wings. A few days after the clinical examination, the animal died naturally. Once submitted to necropsy, histopathological evaluation of the lesions revealed clusters of proliferating epithelial cells expanding toward the dermis. Some of these cells had round, well-defined, intracytoplasmic eosinophilic material suggestive of poxvirus inclusion (Bollinger bodies). PCR performed on the DNA extracted from tissue samples amplified a fragment of the 4b core protein gene (fpv167), which was purified and sequenced. This fragment of Avipoxvirus DNA present in these tumor-like lesions showed high genetic homology (100.0%) with other poxviruses detected in different avian species in several countries, but none of them were related to tumor-like lesions or squamous cell carcinoma. This is the first report of Avipoxvirus detected in tumor-like lesions of a white-faced whistling duck with phylogenetic analysis of the virus.(AU)

Avipoxvirus é o agente etiológico da varíola (bouba) aviária, uma doença bem descrita em aves de cativeiro e selvagens, tendo sido associada a lesões semelhantes a tumores em algumas dessas espécies. Uma marreca piadeira (Dendrocygna viduata), criada em cativeiro, foi atendida em um Hospital Veterinário na região nordeste devido à presença de nódulos cutâneos em ambas as asas. Alguns dias após o exame clínico, o animal veio a óbito naturalmente. A ave foi submetida à necropsia e coletados fragmentos das lesões para análise histopatológica, que revelou proliferação de células epiteliais expandindo para a derme. Algumas dessas células possuíam material eosinofílico intracitoplasmático e bem definido, sugestivo de inclusão de poxvírus (corpúsculos de Bollinger). A PCR realizada a partir do DNA extraído de amostras das lesões amplificou um fragmento do gene da proteína do núcleo 4b (fpv 167), que foi purificado e sequenciado. Esse fragmento de DNA de Avipoxvirus presente nas lesões relevou alta homologia genética (100,0%) com outros poxvírus detectados em diferentes espécies de aves em vários países, mas nenhum deles estava relacionado a lesões tumorais ou carcinoma espinocelular. Este é o primeiro relato de Avipoxvirus detectado em lesões semelhantes a tumores em uma marreca piadeira com caracterização molecular do vírus.(AU)

Detection of avian metapneumovirus subtype A from wild birds in the State of São Paulo, Brazil / Detecção de metapneumovirus aviário subtipo A em aves silvestres no estado de São Paulo, Brasil

The present study investigated the circulation of avian metapneumovirus (aMPV) in wild birds in Brazil. To do so, 131 samples from 366 oropharyngeal or cloacal swabs collected from 18 species of birds were tested individually or in pools by RT-PCR. Samples detected by RT-PCR were selected for DNA sequencing. Thirteen (9.9%) samples were detected by the RT-PCR targeting the N gene and four out of 13 samples were sequenced. Sequencing results showed a high identity with the aMPV subtype A. Our results confirm the circulation of the aMPV subtype A in wild birds in Brazil even five years after its last detection.(AU)

O presente estudo investigou a circulação de metapneumovírus aviário em aves silvestres no Brasil. Para tanto, 131 amostras de 366 suabes orofaringeanos ou cloacais coletados de 18 espécies de aves foram testadas individualmente ou na forma de pools por RT-PCR. As amostras detectadas por RT-PCR foram selecionadas para sequenciamento. Treze (9,9%) das amostras foram detectadas por RT-PCR tendo o gene N como alvo; destas, quatro foram sequenciadas com sucesso. Resultados do sequenciamento mostraram alta identidade com o aMPV de subtipo A. Nossos resultados confirmam a circulação de aMPV subtipo A em aves silvestres no Brasil mesmo cinco anos após sua última detecção.(AU)

Detection of avian metapneumovirus subtype A from wild birds in the State of São Paulo, Brazil / Detecção de metapneumovirus aviário subtipo A em aves silvestres no estado de São Paulo, Brasil

The present study investigated the circulation of avian metapneumovirus (aMPV) in wild birds in Brazil. To do so, 131 samples from 366 oropharyngeal or cloacal swabs collected from 18 species of birds were tested individually or in pools by RT-PCR. Samples detected by RT-PCR were selected for DNA sequencing. Thirteen (9.9%) samples were detected by the RT-PCR targeting the N gene and four out of 13 samples were sequenced. Sequencing results showed a high identity with the aMPV subtype A. Our results confirm the circulation of the aMPV subtype A in wild birds in Brazil even five years after its last detection.(AU)

O presente estudo investigou a circulação de metapneumovírus aviário em aves silvestres no Brasil. Para tanto, 131 amostras de 366 suabes orofaringeanos ou cloacais coletados de 18 espécies de aves foram testadas individualmente ou na forma de pools por RT-PCR. As amostras detectadas por RT-PCR foram selecionadas para sequenciamento. Treze (9,9%) das amostras foram detectadas por RT-PCR tendo o gene N como alvo; destas, quatro foram sequenciadas com sucesso. Resultados do sequenciamento mostraram alta identidade com o aMPV de subtipo A. Nossos resultados confirmam a circulação de aMPV subtipo A em aves silvestres no Brasil mesmo cinco anos após sua última detecção.(AU)

First phylogenetic analysis of Avipoxvirus (APV) in Brazil / Primeira análise filogenética de Avipoxvirus (APV) no Brasil

This study represents the first phylogenetic analysis of avian poxvirus recovered from turkeys in Brazil. The clinical disorders related to fowlpox herein described occurred in a turkey housing system. The birds displaying characteristic pox lesions which were observed on the neck, eyelids and beak of the turkeys. Four affected turkeys were randomly chosen, euthanized and necropsied. Tissues samples were submitted for histopathological analysis and total DNA was further extracted, amplified by conventional PCR, sequenced and phylogenetically analyzed. Avian poxviruses specific PCR was performed based on P4b core protein gene sequence. The histological analysis revealed dermal inflammatory process, granulation tissue, hyperplasia of epithelial cells and inclusion bodies. The P4b gene was detected in all samples. Sequencing revealed a 100% nucleotide and amino acid sequence identity among the samples, and the sequences were deposited in GenBank®. The four Avian poxviruses fragments sequenced in this study clustered along the A1 clade of avipoxviruses, and were classified as Avipoxvirus (APV). Additional studies, such as virus isolation, PCR and sequencing includinga large number of specimens from the Brazilian turkey production must be conducted due to the hazardous risk that poxvirus infections may cause to the Brazilian poultry production scenario, given that Brazil's turkey production attracts attention due to its economic importance worldwide. Our findings point to the need to identify the prevalence of APV in Brazilian turkey production, to perform risk assessment studies and continued surveillance of APV infections in both wild and commercial avian species.(AU)

Este trabalho representa a primeira análise filogenética de Poxvirus aviário detectado em perus no Brasil. Os distúrbios clínicos relacionados com bouba aviária aqui descritos ocorreram em um sistema de alojamento de perus. As aves apresentaram lesões características de varíola observadas no pescoço, pálpebras e bico das aves. Quatro perus com sinais característicos foram escolhidos aleatoriamente, sacrificados e submetidos à autópsia. Amostras de tecido foram submetidas à análise histopatológica e o DNA total foi extraído, amplificado por PCR convencional e os amplicons foram sequenciados e analisados ​​filogeneticamente. A PCR específica para Poxvírus aviário foi realizada com base na seqüência do gene da proteína do núcleo P4b. A análise histológica revelou um processo inflamatório dérmico, tecido de granulação, hiperplasia de células epiteliais e corpúsculos de inclusão. O gene P4b foi detectado em todas as amostras. O sequenciamento revelou uma identidade entre nucleotídeos e aminoácido de 100% entre as amostras e as sequências foram depositadas no GenBank®. Os quatro fragmentos de poxvírus aviário sequenciado neste estudo foram agrupados no clado A1 de avipoxvirus e foram classificados como Avipoxvirus (APV). Estudos adicionais, como isolamento viral, PCR e sequenciamento, incluindo um grande número de perus da produção brasileira devem ser conduzidos devido ao grave risco que a infecção por poxvírus pode causar ao cenário de produção avícola brasileira, tendo em vista que a produção brasileira de perus atrai atenção devido a sua importância mundial. Nossos resultados apontam para a necessidade de identificar a prevalência da APV na produção de peru no Brasil, para realizar estudos de avaliação de risco e continuada monitoração de infecções por APV nas espécies de aves comerciais e silvestres.(AU)

Systematization, distribution and territory of the middle cerebral artery and cerebroethmoidal artery on the brain's surface of the turkey (Meleagris gallopavo) / Sistematização, distribuição e território da artéria cerebral média e cerebroetmoidal na superfície do encéfalo em peru (Meleagris gallopavo)

Background: The cerebral arteries constitute the main blood transportation via to the brain and play a fundamental role in the maintenance of normal cerebral activities. There are a few studies about the encephalic irrigation in the wild and exotic species. This study aims to describe and systematize the middle cerebral artery and cerebroethmoidal artery of Meleagris gallopavo, to determine a standard model and territorial area of irrigation, and the main variations in this species.Materials, Methods & Results: For this research, 30 brain with the cervical spinal cord segment of 10 male and 20 female young and adult turkeys (Meleagris gallopavo) were used. Discarded animals were provided from Doux Frangosul breeding center, located in Caxias do Sul city, State of Rio Grande do Sul, Brazil, 2006. Animals were euthanized with T 61, with an intravenous injection. After the removal of the skin, the sternum was removed, the common carotid arteries were cannulated through the brachycephalic trunks. The arterial system of the 30 animals were rinsed with saline solution with heparin and filled with latex colored in red. Animals remained immersed for one hour in running water, and next, the neck was sectioned. Next, a bone window was opened in the skullcap. Pieces were fixed in 20% formaldehyde for seven days, and after this period, the brain was removed from the cranial vault...(AU)

Systematization, distribution and territory of the middle cerebral artery and cerebroethmoidal artery on the brain's surface of the turkey (Meleagris gallopavo) / Sistematização, distribuição e território da artéria cerebral média e cerebroetmoidal na superfície do encéfalo em peru (Meleagris gallopavo)

Background: The cerebral arteries constitute the main blood transportation via to the brain and play a fundamental role in the maintenance of normal cerebral activities. There are a few studies about the encephalic irrigation in the wild and exotic species. This study aims to describe and systematize the middle cerebral artery and cerebroethmoidal artery of Meleagris gallopavo, to determine a standard model and territorial area of irrigation, and the main variations in this species.Materials, Methods & Results: For this research, 30 brain with the cervical spinal cord segment of 10 male and 20 female young and adult turkeys (Meleagris gallopavo) were used. Discarded animals were provided from Doux Frangosul breeding center, located in Caxias do Sul city, State of Rio Grande do Sul, Brazil, 2006. Animals were euthanized with T 61, with an intravenous injection. After the removal of the skin, the sternum was removed, the common carotid arteries were cannulated through the brachycephalic trunks. The arterial system of the 30 animals were rinsed with saline solution with heparin and filled with latex colored in red. Animals remained immersed for one hour in running water, and next, the neck was sectioned. Next, a bone window was opened in the skullcap. Pieces were fixed in 20% formaldehyde for seven days, and after this period, the brain was removed from the cranial vault...

Reproduction, development and habits of the large turkey louse Chelopistes meleagridis (Phthiraptera: Ischnocera) under laboratory conditions / Reprodução, desenvolvimento e hábitos de Chelopistes meleagridis (Phthiraptera: Ischnocera) em laboratório

The bionomy of Chelopistes meleagridis off the host was observed with the aim of better understanding the aspects of this species' life cycle. For this purpose, C. meleagridis adults were collected and maintained under controlled conditions to reproduce (35°C and RH > 80%), with turkey feathers as the food source. From the offspring of these lice, the development of 150 individuals was observed from the egg to the adult phase. These eggs were divided into two groups of 75 each. After hatching, one group was given a diet composed of feathers while the other received feathers plus skin of the host turkey (Meleagris gallopavo). The “feather + skin” diet resulted in the greatest number of adults, so this diet was given to the next generation of lice reared in vitro, starting from the first instar, to observe their fertility, fecundity and longevity. High reproduction rates were found in relation to other lice of the Ischnocera sub-order, particularly the number of eggs per day and number of eggs produced per female over the lifetime (means of 2.54 and 26.61 eggs, respectively, for wild females and 2.11 and 29.33 eggs for laboratory-reared females). The inclusion of skin in the diet was a determining factor for development to the adult stage, since 48% of the lice fed this diet reached that stage, versus 1.3% that reached maturity fed only with feathers. The development time of the males and females was similar (mean of 29.38 days), without any difference in the sexual proportion of the adults.(AU)

A bionomia de Chelopistes meleagridis fora do hospedeiro foi observada com o objetivo de compreender aspectos relacionados ao ciclo de vida desta espécie. Para isto, adultos de C. meleagridis foram coletados e colocados em condições controladas (temperatura de 35°C e umidade relativa superior a 80%) para se reproduzir, oferecendo-se pena como alimento. Da prole destes adultos, foi observado o desenvolvimento de 150 indivíduos desde o ovo até a fase adulta. Para 75 destes, foi oferecida a dieta composta de pena, enquanto para os outros 75 a dieta foi composta de pena e pele do hospedeiro (peru, Meleagris gallopavo). Ao verificar que a dieta “pena + pele” foi a que resultou no maior número de adultos, foram observadas a fertilidade, fecundidade e a longevidade de piolhos criados in vitro desde o primeiro ínstar alimentados com esta dieta. Valores altos relacionados à reprodução desta espécie foram encontrados em relação a outros piolhos da subordem Ischnocera, destacando-se: número de ovos produzidos por dia e número de ovos produzidos por fêmeas durante a vida (médias de 2,54 e 26,61 ovos, respectivamente, para fêmeas selvagens e 2,11 e 29,33 ovos, respectivamente, para fêmeas criadas in vitro.). A inclusão de pele na dieta foi determinante para o desenvolvimento até o estágio adulto, uma vez que 48% dos piolhos alimentados com essa dieta atingiram a fase adulta. Quando foi oferecido apenas pena, 1,3% dos piolhos atingiram a maturidade. O tempo de desenvolvimento de machos e fêmeas foi semelhante (média de 29,38 dias) sem haver diferença na proporção sexual dos adultos.(AU)

Pathological and Molecular Analysis of Turkey Coronavirus Replication in Captive Wild Turkey Embryos (Broad-Breasted Bronze Breeder)

The degree of genetic and pathologic variation exhibited by a turkey Coronavirus (TCoV) strain was investigated after nine serial passages in 25-day-old turkey embryos obtained from wild broad-breasted bronze breeders. In spite of spleen, liver, kidneys, cloacal bursa and thymus have been collected and analysed, the main histopathological changes were only documented in the intestine sections. Microscopic lesions were characterized as mild enteritis, low degree of enterocyte vacuolization and detachment of the intestinal villous after five consecutive passages and were considered absent in the last passages. Genealogic analysis based on S1 and S2 DNA sequences suggested that Brazilian isolate might be considered as originated from TCoV strains circulating in the United States, as 100% identity with TCoV-Gl strain. Although S1 S2 sequences from each passage revealed no significant point mutations, and no correlation could be speculate between S2 nucleotide changes and pathologic features in infected embryos. This is the first demonstration of wild turkey embryos as a model for TCoV isolation and propagation.(AU)

Response of seeds and pollen of Onobrychis viciifolia and Onobrychis oxyodonta var. armena to NaCl stress

Sainfoin (Onobrychis viciifolia Scop.) is an important forage legume crop with 52 species adapted to dry and poor soils in Turkey, but little is known about the effects of salinity on germination and seedling growth in arid and semiarid regions suffering from salinity problem. The seeds and pollen of two species of sainfoin O. viciifolia and O. oxyodonta var. armena (Syn: O. armena) were exposed to 0, 5, 10, 20 and 30 dS m-1 of NaCl under in vivo and in vitro conditions and evaluated for germination under salt stress by comparing germination percentage, mean germination time, root and shoot length, fresh and dry seedling weight and dry matter. Increased salinity levels generally resulted in decrease in all traits except time to germination, dry seedling weight and dry matter, which increased at high salinity levels. O. viciifolia seeds germinated and grew more rapidly compared to O. armena seeds under NaCl stress. No decrease in germination and seedling growth up to 10 dS m-1 was recorded. On the other hand, there was a clear difference for germination and seedling growth between in vivo and in vitro conditions. Lower values were obtained from in vitro experiments; suggesting that mineral salts, sucrose and agar may have resulted in higher osmotic potential inhibiting germination and seedling growth of species compared in vivo conditions. Decrease in pollen germination with increasing salinities was very sharp, indicating that pollen germination had higher sensitive to salinity. But, pollen grains of O. armena germinated rapidly compared to O. viciifolia. The results emphasize that in vivo experiments could be used for screening of NaCl tolerance in sainfoin cultivars without expensive chemicals and sophisticated equipments, but pollen germination is more appropriate for its wild relatives.

Response of seeds and pollen of Onobrychis viciifolia and Onobrychis oxyodonta var. armena to NaCl stress

Sainfoin (Onobrychis viciifolia Scop.) is an important forage legume crop with 52 species adapted to dry and poor soils in Turkey, but little is known about the effects of salinity on germination and seedling growth in arid and semiarid regions suffering from salinity problem. The seeds and pollen of two species of sainfoin O. viciifolia and O. oxyodonta var. armena (Syn: O. armena) were exposed to 0, 5, 10, 20 and 30 dS m-1 of NaCl under in vivo and in vitro conditions and evaluated for germination under salt stress by comparing germination percentage, mean germination time, root and shoot length, fresh and dry seedling weight and dry matter. Increased salinity levels generally resulted in decrease in all traits except time to germination, dry seedling weight and dry matter, which increased at high salinity levels. O. viciifolia seeds germinated and grew more rapidly compared to O. armena seeds under NaCl stress. No decrease in germination and seedling growth up to 10 dS m-1 was recorded. On the other hand, there was a clear difference for germination and seedling growth between in vivo and in vitro conditions. Lower values were obtained from in vitro experiments; suggesting that mineral salts, sucrose and agar may have resulted in higher osmotic potential inhibiting germination and seedling growth of species compared in vivo conditions. Decrease in pollen germination with increasing salinities was very sharp, indicating that pollen germination had higher sensitive to salinity. But, pollen grains of O. armena germinated rapidly compared to O. viciifolia. The results emphasize that in vivo experiments could be used for screening of NaCl tolerance in sainfoin cultivars without expensive chemicals and sophisticated equipments, but pollen germination is more appropriate for its wild relatives.

Pathological and molecular analysis of turkey Coronavirus replication in captive wild turkey embryos (broad-breasted bronze breeder)

The degree of genetic and pathologic variation exhibited by a turkey Coronavirus (TCoV) strain was investigated after nine serial passages in 25-day-old turkey embryos obtained from wild broad-breasted bronze breeders. In spite of spleen, liver, kidneys, cloacal bursa and thymus have been collected and analysed, the main histopathological changes were only documented in the intestine sections. Microscopic lesions were characterized as mild enteritis, low degree of enterocyte vacuolization and detachment of the intestinal villous after five consecutive passages and were considered absent in the last passages. Genealogic analysis based on S1 and S2 DNA sequences suggested that Brazilian isolate might be considered as originated from TCoV strains circulating in the United States, as 100% identity with TCoV-Gl strain. Although S1 S2 sequences from each passage revealed no significant point mutations, and no correlation could be speculate between S2 nucleotide changes and pathologic features in infected embryos. This is the first demonstration of wild turkey embryos as a model for TCoV isolation and propagation.