Acute infection with Platynosomum illiciens (Trematoda: Dicrocoeliidae) as a clinically relevant and potentially fatal disease in Falco sparverius (Aves: Falconidae) in Brazil.

Platynosomum illiciens is a dicrocoeliid trematode from the biliary tract of warm-blooded vertebrates (felines, primates, marsupials, and birds) reported in different parts of the world. Although the veterinary relevance of platynosomosis in mammals, especially in domestic felines, has been increasingly evidenced in the scientific literature, studies involving avian disease caused by P. illiciens are comparatively scarce. In the present study, a female specimen of the American kestrel, Falco sparverius L., found dead, in November 2019, in Brazil, was necropsied. Parietal biliary effusion in the celomatic cavity was observed, suggesting biliary transudation and gallbladder stasis, which possibily preceded the distension and rupture of gallbladder noted during necropsy. In the microscopic analysis of the bile content, small trematodes were found and characterized as immature stages of Platynosomum after the morphological study. Partial sequences of the cox-1 gene enabled the identification of P. illiciens, with 100% similarity with previously sequenced sympatric isolates from mammals. The finding of immature specimens in a ruptured gallbladder strongly suggests a role for the parasite in biliary flow dysfunction, indicating acute platynosomosis as a clinically relevant and potentially fatal condition that has not yet been discussed.

NEW RECORDS OF NEMATODES IN THE YELLOW-BILLED CUCKOO COCCYZUS AMERICANUS (CUCULIFORMES: CUCULIDAE) BASED ON A MIGRANT SPECIMEN FOUND IN BRAZIL.

The yellow-billed cuckoo, Coccyzus americanus, is a threatened passerine bird native to North America that migrates to overwinter in South America. Although migratory birds have drawn broad attention, given their assumed role in the long-distance dispersal of parasites, studies on the helminth fauna of this cuculid are scarce. In the present study, nematodes found in the gastrointestinal tract of a specimen of C. americanus found in Belo Horizonte, State of Minas Gerais, southeastern Brazil, were characterized morphologically. Five species of gastrointestinal nematodes belonging to 4 families were identified: Synhimantus (Dispharynx) nasuta (Rudolphi, 1819) and Synhimantus (Dispharynx) resticulaCanavan, 1929 (Acuariidae), Microtetrameres sp. (Tetrameridae), and Cyrnea piayaeSandground, 1929 (Habronematidae) and Subulura halli Barreto, 1918 (Subuluridae). Except for Microtetrameres sp., all other nematodes are reported in this host for the first time. Although it is difficult to accurately determine the geographical origin of infections, it is important to note that Sy. (D.) resticula and Su. halli are reported for the first time in Brazil. Additionally, the absence of the former species infecting hosts where the widespread and generalist Sy. (D.) nasuta was found may indicate that Sy. (D.) resticula have either been overlooked in previous studies or that this nematode is rare indeed. Finally, the geographical distribution of Su. halli is greatly expanded and now includes the Americas. Aspects related to the dispersion of these parasites, both in migrating and native hosts, are briefly discussed.

Identification of the avian tracheal trematode Typhlocoelum cucumerinum (Trematoda: Cyclocoelidae) in a host-parasite-environment system: diagnosis, life cycle and molecular phylogeny.

Typhlocoelum cucumerinum is a tracheal parasite of birds widely distributed across the globe. Nevertheless, aspects of the biology of this cyclocoelid are still poorly understood. Herein, we report the finding of T. cucumerinum in definitive and intermediate hosts from an urban waterbody of Brazil. The parasite was initially detected during the necropsy of domestic Muscovy ducks (Cairina moschata) found dead in the locality. Coproparasitological tests in live animals revealed that 12/47 (25.53%) Muscovy ducks and 2/8 (25%) mallards (Anas platyrhynchos platyrhynchos) were infected with T. cucumerinum. Moreover, rediae and metacercariae morphologically similar to T. cucumerinum were found in 3/248 (1.33%) Biomphalaria straminea collected in the same waterbody frequented by the birds. The conspecificity between the adult and the larval stages was confirmed molecularly (100% similarity in Cox-1). Moreover, the phylogenetic position of T. cucumerinum was determined for the first time based on partial fragments of the 28S, Cox-1 and Nad-1 genes. The species grouped with other members of the subfamily Typhlocoelinae with sequences available, but the data obtained do not support the distinctiveness of the genera Typhlocoelum and Tracheophilus. Further studies involving a broader range of species can result in taxonomic rearrangements in Typhlocoelinae.

Polyostotic osteosarcoma associated with avian leukosis virus infection in a captive bare-faced curassow (Crax fasciolata).

BACKGROUND: Osteosarcoma is a malignant mesenchymal bone tumor. Although it is a common tumor in the appendicular skeleton of dogs and cats, it is rarely reported in birds. Retroviruses are usually associated with solid tumor development in different avian species. CASE PRESENTATION: This report aims to describe a case of osteosarcoma associated with the avian leukosis virus in a captive bare-faced curassow (Crax fasciolata). A captive adult female bare-faced curassow presented with lameness, hyporexia, and a non-ulcerative and firm tumor in the right femur. The bird was euthanized due to the poor prognosis. Histopathology revealed an infiltrative mesenchymal neoplasm consisting of spindle cells with moderate cell pleomorphism, organized in bundles and interspersed by marked deposition of the osteoid matrix, which was compatible with osteosarcoma affecting both femur and tibiotarsus, with renal metastasis. Immunohistochemistry of the primary and metastatic tumor demonstrated vimentin expression by neoplastic cells. Samples of the neoplasm, bone marrow, and spleen were processed for PCR, which enabled the demonstration of proviral avian leukosis virus (ALV) DNA. CONCLUSIONS: To the best of our knowledge, this is the first report of an osteosarcoma in a bare-faced curassow with an unusual polyostotic manifestation and associated with ALV infection.

Evaluation of actions of the official veterinary service to mitigate outbreaks of infectious laryngotracheitis and improve biosecurity on laying hen farms

ABSTRACT: Infectious laryngotracheitis (ILT), caused by an Alphaherpesvirus (Gallid herpesvirus-1; GaHV-1), has been noticed in the region of the Terras Altas da Mantiqueira, Minas Gerais. From 2010 to 2018, the Serviço Veterinário Oficial (SVO) of the Instituto Mineiro Agropecuário (IMA), implemented measures to prevent spread of the virus to other regions and control the disease in the area. Due to the close proximity and consequent epidemiological link among farms, the region was considered a unique epidemiological unit. To check the efficiency of the ILT control measures, we carried out: (1) a seroepidemiological survey, (2) questionnaires for evaluating biosecurity measures; and (3) an evaluation of the influence of farm population density on the occurrence of ILT. In 2016, 2017, and 2018, ILT was investigated using epidemiological and clinicopathological methods, along with GaHV-1 molecular detection. Serological survey was carried out on 24 farms in the quarantined region and on 13 farms from other regions of the state. In 2010 and 2018, questionnaires were applied to collect data and determine indicators of biosecurity practices in all farms of the quarantined area. The differences were then assessed (Wilcoxons p 0.05). The results indicated positive serology throughout the region, although only on four farms (16.6%) the chickens have clinical signs, macroscopic and histological lesions of ILT. The prevalence of viral infection increased from 2016 (27%) to 2017 (50%) and was higher in farms with a high stock density (p=0.033). No disease, virus or antibodies were detected in the farms outside of the quarantined area. Although the biosecurity indicators had improved on all farms in the quarantined area (p 0.05), the virus was active and circulating in the region. The contingency measures have contained the outbreak, but biosecurity practices are paramount in the control of new outbreaks. Official control will be maintained in the region, including surveillance of new cases and biosecurity procedures to mitigate the risk of the virus reaching other regions.

RESUMO: Laringotraqueíte infecciosa (LTI), causada por um alfaherpesvírus (herpesvírus Gallid-1; GaHV-1), foi observada na região das Terras Altas da Mantiqueira, Minas Gerais. De 2010 a 2018, o Serviço Veterinário Oficial (SVO) do Instituto Mineiro Agropecuário (IMA) implementou medidas para impedir a disseminação do vírus para outras regiões do estado e controlar a doença na região interditada. Devido à proximidade e consequente vínculo epidemiológico entre as granjas, a região foi considerada uma unidade epidemiológica única. Para verificar a eficiência das medidas de controle de LTI, foram realizados: (1) pesquisa soroepidemiológica, (2) questionários para avaliar medidas de biosseguridade; e (3) avaliação da influência da densidade populacional da granja na ocorrência de LTI. Em 2016, 2017 e 2018, a LTI foi investigada usando métodos epidemiológicos e clínico-patológicos, com a detecção molecular de GaHV-1. O levantamento sorológico foi realizado em 24 granjas da região interditada e em 13 granjas de outras regiões do estado. Em 2010 e 2018, foram aplicados questionários para coletar dados e determinar indicadores de medidas de biosseguridade em todas as granjas da área interditada. As diferenças foram avaliadas (p 0,05 de Wilcoxon). Os resultados indicaram sorologia positiva em toda a região, embora apenas em quatro granjas (16,6%) as galinhas apresentaram sinais clínicos, lesões macroscópicas e histológicas da LTI. A prevalência de infecção viral aumentou de 2016 (27%) para 2017 (50%) e foi maior em fazendas com alta densidade de alojamento (p=0,033). Presença da doença, vírus ou anticorpos foram detectados nas granjas fora da área interditada. Embora os indicadores de biosseguridade tenham melhorado em todas as fazendas da área interditada (p 0,05), o vírus está ativo e circulava na região. As medidas de contingência contiveram o surto, mas as práticas de biosseguridade são fundamentais para o controle de novos surtos. O controle oficial será mantido na região, incluindo a vigilância de novos casos e procedimentos de biosseguridade para mitigar o risco de transmissão do vírus para outras regiões.

Evaluation of actions of the official veterinary service to mitigate outbreaks of infectious laryngotracheitis and improve biosecurity on laying hen farms / Avaliação de ações do serviço veterinário oficial para bloqueio de foco de laringotraqueite infecciosa e melhoria da biosseguridade em granjas de postura

Infectious laryngotracheitis (ILT), caused by an Alphaherpesvirus (Gallid herpesvirus-1; GaHV-1), has been noticed in the region of the Terras Altas da Mantiqueira, Minas Gerais. From 2010 to 2018, the "Serviço Veterinário Oficial" (SVO) of the "Instituto Mineiro Agropecuário" (IMA), implemented measures to prevent spread of the virus to other regions and control the disease in the area. Due to the close proximity and consequent epidemiological link among farms, the region was considered a unique epidemiological unit. To check the efficiency of the ILT control measures, we carried out: (1) a seroepidemiological survey, (2) questionnaires for evaluating biosecurity measures; and (3) an evaluation of the influence of farm population density on the occurrence of ILT. In 2016, 2017, and 2018, ILT was investigated using epidemiological and clinicopathological methods, along with GaHV-1 molecular detection. Serological survey was carried out on 24 farms in the quarantined region and on 13 farms from other regions of the state. In 2010 and 2018, questionnaires were applied to collect data and determine indicators of biosecurity practices in all farms of the quarantined area. The differences were then assessed (Wilcoxon's p<0.05). The results indicated positive serology throughout the region, although only on four farms (16.6%) the chickens have clinical signs, macroscopic and histological lesions of ILT. The prevalence of viral infection increased from 2016 (27%) to 2017 (50%) and was higher in farms with a high stock density (p=0.033). No disease, virus or antibodies were detected in the farms outside of the quarantined area. Although the biosecurity indicators had improved on all farms in the quarantined area (p<0.05), the virus was active and circulating in the region. The contingency measures have contained the outbreak, but biosecurity practices are paramount in the control of new outbreaks. Official control will be maintained in the region, including surveillance of new cases and biosecurity procedures to mitigate the risk of the virus reaching other regions.(AU)

Laringotraqueíte infecciosa (LTI), causada por um alfaherpesvírus (herpesvírus Gallid-1; GaHV-1), foi observada na região das Terras Altas da Mantiqueira, Minas Gerais. De 2010 a 2018, o Serviço Veterinário Oficial (SVO) do Instituto Mineiro Agropecuário (IMA) implementou medidas para impedir a disseminação do vírus para outras regiões do estado e controlar a doença na região interditada. Devido à proximidade e consequente vínculo epidemiológico entre as granjas, a região foi considerada uma unidade epidemiológica única. Para verificar a eficiência das medidas de controle de LTI, foram realizados: (1) pesquisa soroepidemiológica, (2) questionários para avaliar medidas de biosseguridade; e (3) avaliação da influência da densidade populacional da granja na ocorrência de LTI. Em 2016, 2017 e 2018, a LTI foi investigada usando métodos epidemiológicos e clínico-patológicos, com a detecção molecular de GaHV-1. O levantamento sorológico foi realizado em 24 granjas da região interditada e em 13 granjas de outras regiões do estado. Em 2010 e 2018, foram aplicados questionários para coletar dados e determinar indicadores de medidas de biosseguridade em todas as granjas da área interditada. As diferenças foram avaliadas (p<0,05 de Wilcoxon). Os resultados indicaram sorologia positiva em toda a região, embora apenas em quatro granjas (16,6%) as galinhas apresentaram sinais clínicos, lesões macroscópicas e histológicas da LTI. A prevalência de infecção viral aumentou de 2016 (27%) para 2017 (50%) e foi maior em fazendas com alta densidade de alojamento (p=0,033). Presença da doença, vírus ou anticorpos foram detectados nas granjas fora da área interditada. Embora os indicadores de biosseguridade tenham melhorado em todas as fazendas da área interditada (p<0,05), o vírus está ativo e circulava na região. As medidas de contingência contiveram o surto, mas as práticas de biosseguridade são fundamentais para o controle de novos surtos. O controle oficial será mantido na região, incluindo a vigilância de novos casos e procedimentos de biosseguridade para mitigar o risco de transmissão do vírus para outras regiões.(AU)

Haemoproteus paraortalidum n. sp. in captive Black-fronted Piping-guans Aburria jacutinga (Galliformes, Cracidae): High prevalence in a population reintroduced into the wild.

Haemosporidian parasites of the genus Haemoproteus are widespread and can cause disease and even mortality in birds under natural and captive conditions. The Black-fronted Piping-guan (Aburria jacutinga) is an endangered Neotropical bird of the Cracidae (Galliformes) going through a reintroduction program to avoid extinction. We used microscopic examination and partial cytochrome b DNA sequencing to describe a new Haemoproteus species infecting Black-fronted Piping-guans bred and raised in captivity that were reintroduced into the Atlantic rainforest. Haemoproteus (Parahaemoproteus) paraortalidum n. sp. was detected in the blood of 19 out of 29 examined birds. The new species is distinguished from other haemoproteids due to the shape of gametocytes, which have pointed ends in young stages, and due to the presence of vacuole-like unstained spaces in macrogametocytes and numerous volutin granules both in macro- and microgametocytes. Illustrations of the new species are provided. Phylogenetic inference positioned this parasite in the Parahaemoproteus subgenus clade together with the other two Haemoproteus genetic lineages detected in cracids up to date. We discuss possible implications of the reintroduction of birds infected with haemosporidian parasites into the wild. Treatment of Haemoproteus infections remains insufficiently studied, but should be considered for infected birds before reintroduction to improve host reproductive and survival rates after release.

Experimental avian philophthalmosis: Evaluation of diagnosis and treatment of chickens infected with Philophthalmus gralli (Trematoda: Philophthalmidae).

The trematodes of the genus Philophthalmus are eye flukes that cause damage to ocular structures of animals and humans. Despite the increasing number of cases reported in birds, studies related to the diagnosis of subclinical philophthalmosis are lacking, and there are no effective therapeutic regimens available. In the present study, we evaluated the diagnosis and treatment of philophthalmosis in specific pathogen-free chickens (Gallus gallus domesticus) experimentally infected with Philophthalmus gralli. Four chickens were inoculated with metacercariae of P. gralli (20 per eye) obtained from cercariae emerged from naturally infected Melanoides tuberculata. From 90 days post-infection, the chickens were subjected to direct ophthalmic examination (DOE) and conjunctival sac lavage (CSL). The latter technique consisted of lavage of each eye with 200 µL sterile saline solution and subsequent microscopical examination of the collected fluid for the presence of eggs of P. gralli. The anthelminthic treatment protocols included praziquantel (PZQ) at 10, 50, or 100 mg/kg (single dose given intramuscularly), or fenbendazole (FBZ) at 50 mg/kg (three doses at 24 h-intervals given per os). The treatment protocols were performed at 14 day-intervals between each dosage of PZQ. Chickens developed a minimum of one to more than five adult P. gralli per eye, except for one chicken that had a single eye with one parasite. No ocular clinical signs or changes in behavior were noted in any chickens. DOE and CSL were considered techniques with similar sensitivity for the diagnosis of avian philophthalmosis. The data suggested that PZQ and FBZ, at the dosages and schedules employed, are not effective for the complete elimination of P. gralli. CSL is proposed as a complementary technique for the diagnosis and monitoring of philophthalmosis post-treatment, especially in subclinical cases. The evaluation of new protocols, routes of administration, and anthelmintic drugs are needed for successful pharmacological treatment of philophthalmosis.

Antitoxin therapy of natural avian botulism outbreaks occurred in Brazil.

Botulism commonly affects water birds and it has recently been observed to be emerging in poultry production. In the present work, outbreaks of botulism in wild native species, such as the black-fronted Piping-guan (Aburria jacutinga), wild duck (Cairina moschata) and its crosses with mallard (Anas platyrhynchos), and domestic chickens (Gallus gallus domesticus) are described. Following treatments with a commercial botulism antitoxin CD, 28 (96.5%) out of 29 animals fully recovered after 24-72 h. The antitoxin therapy was shown to be a useful option for the treatment of affected birds, including those that were severely affected.

Avian Pox in Native Captive Psittacines, Brazil, 2015.

To investigate an outbreak of avian pox in psittacines in a conservation facility, we examined 94 birds of 10 psittacine species, including sick and healthy birds. We found psittacine pox virus in 23 of 27 sick birds and 4 of 67 healthy birds. Further characterization is needed for these isolates.

Investigação epidemiológica de estomatite vesicular por achados clínicos em bovinos e equinos no Estado do Maranhão / Stomatitis vesicular in cattle and horse in the Maranhão state

A Estomatite Vesicular (EV) é uma doença infecciosa que acomete equinos, bovinos, suínos, mamíferos silvestres e humanos. Por apresentar sinais clínicos semelhantes a outras doenças vesiculares, principalmente, febre aftosa, sua presença em determinadas regiões pode interferir no intercâmbio comercial internacional dos animais, seus produtos e subprodutos. Apesar de sua importância, a epidemiologia e a manutenção do vírus no ambiente não estão totalmente esclarecidas dificultando a aplicação de medidas de controle efetivas. A doença já foi diagnosticada em todas as regiões brasileiras. Bovinos com sialorréia, perda do epitélio lingual, lesões abertas com bordas amareladas nas gengivas, lábios, língua e mucosa oral e equinos com sialorréia e lesões abertas na mucosa oral e lábios foram observados e notificados ao Serviço Veterinário Oficial do Estado do Maranhão, Agência Estadual de Defesa Agropecuária do Maranhão (AGRD/MA). Amostras de soro de equinos e bovinos com sintomas de EV foram coletadas para investigação por ELISA e por neutralização viral, além do diagnóstico diferencial para Febre Aftosa (FA). Fragmentos epiteliais de bovinos com lesões na língua foram coletados para identificação molecular do agente. Todos os animais foram negativos para FA. Todos os bovinos e equinos foram reativos para EV nos testes sorológicos. A partir dos fragmentos epiteliais de bovinos enviados ao Instituto Biológico de São Paulo para PCR, foi possível caracterizar o agente como VesiculovirusIndiana III (Alagoas/VSAV).

Vesicular stomatitis (VS) is an infectious viral disease that affects bovines, equines, swine, wild animals and humans. As it is indistinguishable from other vesicular diseases, mainly Foot and Mouth Disease (FMD), it causes restrictions in commercial livestock trade at national and international levels and also significant economic losses. As the epidemiology and maintenance of VS virus in nature are not clearly understood it is difficult to take effective control measures. VS was diagnosed in some regions of Brazil, such as Minas Gerais, Santa Catarina, São Paulo and Alagoas. Cattle and horses with clinical symptoms of drooling, shedding of the lingual epithelium, presence of vesicles on the oral mucosa were observed and reported to the National Animal Health Office health of Maranhão State, Brazil. Samples of serum of these animals were collected and sent to Laboratório Nacional de Agropecuaria for ELISA and virus neutralization and differential diagnosis for Foot and Mouth Disease (FMD). The results of ELISA confirmed the VS. In the differential diagnosis, the results were negative for FMD. Samples of bovine epithelial tissue for VS by PCR confirmation of diagnosis were collected and sent to Biological Institute of São Paulo. Molecular results confirmed the VesiculovirusIndiana III (Alagoas/VSAV) infection.

Seleção, caracterização e clonagem dos genes fljB e groEL agonistas dos receptores de reconhecimento de padrão do sistema imune inato das aves / Selection, characterization and cloning of the fljB and groEL genes agonists of the innate immune system pattern recognition receptors of birds

The recombinant production of innate immune system pattern recognition receptor agonists has provided a new tool for the production of immunostimulants for animals. The molecular pattern associated with the pathogen (PAMP), flagellin, coded by the fljB gene from Salmonella Typhimirium, and the molecular pattern associated to the damage (DAMP), HSP60, coded by the groEL gene from S. Typhimurium and S. Enteritidis, are recognized by pattern recognition receptors (PRRs) of the innate immune system of birds. In the present study, we performed the cloning of genetic fragments of the genes fljB, from S. Typhimurium, and groEL from S. Typhimurium and S. Enteritidis inserted in expression vector pET100/D-TOPO and transformed in E. coli TO10 cells. The clones were evaluated by colony PCR, plasmidial DNA PCR and genome sequencing in order to confirm the presence of these genes. In the colony PCR, we identified the presence of genes groEL (S. Enteritidis), groEL (S. Typhimurium) and fljB (S. Typhimurium) in 80%, 60% and 80% of the transformed colonies, respectively. The cloning system adopted allowed the production of HSP60 genetic fragment clones and flagellin of Salmonella strains, allowing the posterior use of these clones in gene expression trials, with the future potential of being used as non-specific immunostimulants for birds.

A produção recombinante de agonistas dos receptores do reconhecimento de padrão do sistema imune inato tem fornecido uma nova ferramenta para a produção de imunoestimulantes para animais. O padrão molecular associado ao patógeno (PAMP), flagelina, codificado pelo gene fljB de Salmonella Typhimurium e o padrão molecular associado ao dano (DAMP) HSP60, codificado pelo gene groEL da S. Typhimurium e S. Enteritidis, são reconhecidos por receptores de reconhecimento de padrões (RRPs) do sistema imune inato das aves. No presente estudo, foi feita a clonagem de fragmentos genéticos dos genes fljB de S. Typhimurium e groEL de S. Typhimurium e S. Enteritidis inseridos no vetor de expressão pET100/D-TOPO e transformados em células de E. coli TOP10. Os clones foram avaliados pela PCR de colônia, PCR de DNA plasmidial e sequenciamento genômico para a confirmação da presença desses genes. Na PCR de colônia, foram identificadas em 80%, 60% e 80% das colônias transformadas, a presença dos genes groEL (S. Enteritidis), groEL (S. Typhimurium) e fljB (S. Typhimurium) respectivamente. O sistema de clonagem adotado possibilitou a produção de clones dos fragmentos genéticos da HSP60 e flagelina das cepas de Salmonella, permitindo a utilização posterior desses clones em ensaios de expressão gênica, com potencial futuro de serem utilizados como imunoestimulante inespecífico das aves.

Interactions of Plasmodium juxtanucleare and chicken anaemia virus: establishing a model.

The pathogens Plasmodium juxtanucleare and chicken anaemia virus (CAV) are easily transmitted and potentially harmful to chickens. In this study, we established an experimental model to investigate the effects of avian malaria caused by P. juxtanucleare in white leghorn specific-pathogen-free (SPF) chicks previously immunosuppressed with CAV. Parasitaemia, haematological variables and clinical and pathological parameters were determined in four different experimental groups: chicks coinfected by CAV and P. juxtanucleare strain (Coinfected group), chicks exclusively infected by CAV (CAV group) or P. juxtanucleare (Malaria group) and uninfected chicks (Control group). Our data demonstrated that P. juxtanucleare parasitaemia was significantly higher in the Coinfected group. Furthermore, haematological parameters, including the RBC, haematocrit and haemoglobin concentration were significantly reduced in coinfected chicks. In agreement with the changes observed in haematological features, the mortality among coinfected chicks was higher compared with animals with single infections. Clinical analysis indicated moderate changes related to different organs size (bursa of Fabricius, heart and liver) in coinfected birds. The experimental coinfection of SPF chickens with P. juxtanucleare and CAV may represent a research tool for the study of avian malaria after CAV immunosuppression, enabling measurement of the impacts caused by different pathogens during malarial infection.

Outbreak of infectious laryngotracheitis in large multi-age egg layer chicken flocks in Minas Gerais, Brazil / Surto de laringotraqueíte infecciosa em granjas de galinhas poedeiras de múltiplas idades em Minas Gerais, Brasil

A recent (November 2010) outbreak of infectious laryngotracheitis (ILT) in a multi-age laying hen facility in Minas Gerais state, Brazil, is described. Previous ILT outbreak in laying hens was only notified in São Paulo state, Brazil, in 2002. In the outbreak described here, the affected population was approximately eight million hens, with flock sizes ranging from 100,000 to 2,900,000 chickens. The average mortality ranged from 1 to 6%, and morbidity was around 90% (most of the twenty seven farms of the area were positive for ILT virus). Three multi-age laying farms from one company were selected for this report. Clinical signs included prostration, dyspnea, conjunctivitis, occasional swelling of the paranasal sinuses and bloody mucous nasal discharge. Severely affected chickens presented with dyspnea, gasping and became cyanotic before death. At necropsy, these chickens had fibrinous exudate blocking the larynx and the lumen of cranial part of the trachea. In addition, conjunctivitis with intense hyperemia, edema and sinuses with caseous exudate were present. On histopathology, there were marked necrosis and desquamation of respiratory ephitelium and conjunctiva with numerous syncytial cells formation and fibrinous exudate. Moderate to marked non suppurative (especially lymphocytes and plasma cells) infiltration in the lamina propria also was observed. Sixteen out of 20 examined chickens, eosinophilic intranuclear inclusion bodies were observed in the syncytial cells. The DNA extracted from larynx and trachea produced positive PCR results for ILT virus (ILTV) DNA using formalin-fixed, paraffin embedded (FFPE) samples. Amplicons from a small region of ICP4 gene were submitted to sequencing and showed 100% identity with ILTV EU104910.1 (USA strain), 99% with ILTV JN596963.1 (Australian strain) and 91% with ILTV JN580316.1 (Gallid herpesvirus 1 CEO vaccine strain) and JN580315.1 (Gallid herpesvirus 1 TCO vaccine strain).

Um surto recente (Novembro de 2010) de laringotraqueite infecciosa (LTI) em granjas de postura de múltiplas idades em Minas Gerais, Brasil, é descrito. Um surto de LTI em galinhas de postura havia sido previamente relatado apenas no Estado de São Paulo em 2002. No surto aqui descrito, a população afetada foi de aproximadamente oito milhões de galinhas, com lotes variando de 100.000 a 2.900.000 galinhas. A mortalidade média variou de 1 a 6% e a morbidade atingiu cerca de 90% (a maioria das 27 granjas foram positivas para o virus da LTI). Três granjas com aves de múltiplas idades pertencentes a uma empresa foram selecionadas para o presente relato. Os sinais clinicos incluíram prostração, dispneia, conjuntivite, edema ocasional dos seios paranasais e secreção nasal mucosa e/ou sanguinolenta. As aves severamente afetadas apresentaram acentuada dispneia, aparente engasgo e tornaram-se cianóticas antes da morte. Nestas aves, exsudato fibrinoso denso obstruindo o lúmen da laringe e parte cranial da traqueia foi observado na necropsia. Havia também, conjuntivite com hiperemia intensa e edema, além de sinusite com exsudato caseoso. Na histopatologia, observaram-se necrose e descamação acentuada do epitélio respiratório e da conjuntiva com formação de numerosos sincícios e exsudato fibrinoso. Além disso, infiltrado inflamatório mononuclear (especialmente linfócitos e plasmócitos) moderado a acentuado na lâmina própria foi observado. Corpúsculos de inclusão intranucleares nas células sinciciais foram observados em 16 das 20 aves examinadas. Resultados positivos pela PCR para o virus da LTI foram obtidos de DNA extraído das laringes e traqueias utilizando amostras fixadas em formol e incluidas na parafina. O produto amplificado de uma região pequena do gen ICP4 foi submetido ao sequenciamento e quando comparado com outras sequências depositadas no Genbank mostrou os seguintes resultados: 100% de identidade com uma estirpe do virus de LTI dos Estados Unidos (JN596963.1), 99% de identidade com uma estirpe Australiana e 91% com a estirpe vacinal CEO (JN580316.1) e TCO (JN580315.1).

Fatal coccidiosis by Isospora icterus (Upton & Whitaker, 2000) in captive Campo Troupial (Icterus jamacaii) in Brazil.

An outbreak of coccidiosis by Isospora icterus (I. icterus, Upton & Whitaker, 2000) in captive Campo Troupial (Icterus jamacaii) (Gmelin, 1788) at the Wild Animals Triage Center (IBAMA, Belo Horizonte, Brazil) is described. Clinical history and the necropsy findings documented diarrhea with diffuse necrotic enteritis. Sporulated oocysts (n = 100) had a bilayered wall, were subspherical, and measured 30.1 (27.5-32.5) microm in length and 28.5 (26.2-30.0) microm in width. A polar body but no micropyle was present and the length/width ratio was 1.1 (1.00-1.2). Each oocyst contained two ellipsoidal sporocysts measuring 17.6 (15.0-20.0) microm in length and 12.9 (12.5-15.0) microm in width, with a length/width ratio of 1.4 (1.2-1.5), and with Stieda and sub-Stieda bodies. Each sporocyst contained four sporozoites with granular sporocyst residuum. Oocysts were compatible with those from I. icterus, previously described in Campo Troupial.

Molecular analysis of Brazilian infectious bronchitis field isolates by reverse transcription-polymerase chain reaction, restriction fragment length polymorphism, and partial sequencing of the N gene.

Molecular analysis of 15 Brazilian infectious bronchitis virus (IBV) isolates, obtained from clinical outbreaks of the disease in chickens (broilers or layers) in the state of Minas Gerais (Brazil) between 1972 and 1989, is reported. Using the N protein gene as target, IBVs were analyzed by reverse transcription-polymerase chain reaction/restriction fragment length polymorphism (RT-PCR/RFLP) with the restriction enzymes AvaII, HphI, Sau96I, and Tsp509I and cDNA sequencing. Results obtained from those isolates were compared to 19 sequences available in GenBank. N gene RFLP profiles, cDNA sequences, and predicted amino acid composition were used for the construction of dendrograms. Brazilian isolates were grouped into one distinct group. Identity of predicted N protein amino acid composition varied from 45% (between isolates G and 208) up to 99% (PM 1 and PM2), and, when compared to the other IBVs, the amino acid identity was from 42% (Q3/88 and G) up to 97% (D41 and PM1). The great genetic diversity was shown to occur before the official use of vaccination in Brazil and has remained thereafter.