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Psittaciform orthobornaviruses are currently considered to be a major threat to the psittacine bird population worldwide. Parrot bornavirus (PaBV) was identified recently in Brazil and, since then, few studies have been conducted to understand the epidemiology of PaBV in captive psittacine birds. In the present study, natural infections by PaBV in South American parrots were investigated in two breeding facilities: commercial (A) and conservationist (B). Thirty-eight psittacine of 21 different species were presented for postmortem examination. Tissue samples were collected and investigated for the presence of PaBV-RNA using RT-PCR. In addition, clinical information about these birds was used when available. PaBV infection was detected in 73.7% of all birds investigated, indicating a wide dissemination of this virus in both facilities. From birds investigated in aviary A, 66.7% showed clinical signs, 100% had typical lesions of proventricular dilatation disease (PDD), 100% had mild to severe proventricular dilatation and 88.9% were PaBV-positive. In birds from aviary B, 27.6% showed clinical signs, 65.5% had typical lesions of PDD, 62% had mild to severe proventricular dilatation and 69% were PaBV-positive. Neurological disease was observed more frequently than gastrointestinal disease. Sequencing analysis of the matrix gene fragment revealed the occurrence of genotype 4 (PaBV-4) in both places. About 15.8% of birds in this study are threatened species. We discussed the difficulties and challenges for controlling viral spread in these aviaries and implications for South American psittacine conservation. These results emphasize the urgent need to develop a national regulatory and health standard for breeding psittacine birds in the country.

Native and exotic avian species can act as reservoirs of pathogens, including bacteria and viruses, with conservation and public health implications. A retrospective study on the diagnosis and frequency of diseases in wild and exotic avian species was conducted. The occurrence of particular diseases was associated with the type of captivity or the bird’s origin. The investigation included macroscopic and microscopic descriptions and the molecular determination of the causative agent(s). Additional immunohistochemical (IHC) analysis, PCR, and genetic sequencing were conducted. A total of 243 cases were compiled for the study, mainly consisting of native wild species (39.1%) obtained from illegal trade. Primary infectious diseases, mainly parasitic (18.1%) and viral (17.7%), were the most common, although coinfections were substantial (18.1%) in birds rescued from trafficking. Fractures and neoplasms accounted for 3.7% and 3.3% of the cases, respectively. Parasitic and viral diseases were the most common in both exotic and wild birds. Chlamydia psittaci, a lethal and zoonotic bacterium, was an important cause of death, especially in native Psittaciformes. The recent detection of Psittacid alphaherpesvirus 5 (PsAHV 5) in exotic psittacines and the diagnosis of coinfections in trafficked birds highlight the importance of monitoring avian health to control potential pathogens that may endanger conservation efforts.

Fowl aviadenovirus (FAdV) is an important pathogen in the global poultry industry and the etiology of inclusion body hepatitis-hydropericardium syndrome (HHS) in chickens. Since the 1990s, several outbreaks of HHS have occurred in poultry producing areas, including South America. The coinfection of FAdV and chicken anemia virus (CAV) may markedly impact the incidence of HHS. This study describes an outbreak of HHS in coinfection with CAV in industrial broiler breeders and characterizes the FAdV isolate. The three-week-old male broiler breeders had pale bone marrow, enlarged and yellowish liver, splenomegaly, and atrophied thymus; one chicken was also found with hydropericardium. Virus isolation was performed in SPF chicken embryos of liver and thymus. Tissues of the naturally infected chickens and the inoculated embryos were evaluated by PCR and histopathology. All affected chickens and inoculated embryos were positive for FAdV and CAV. The inoculated embryos had enlarged, greenish and hemorrhagic livers, and 30% died within 7 days of inoculation. Phylogenetic analysis of the FAdV isolate hexon gene partial sequence enabled grouping with E species. The E species has recently become a relevant species in several countries. The association of FAdV with CAV in breeders is of further concern due to both being capable of vertical transmission. Within the last decade, a worldwide upsurge of HHS in broiler breeders owing to failed biosecurity has occurred. In this episode, the failure on biosecurity may have enabled challenge with both FAdV and CAV, with pathological synergism. The CAV-impaired adaptive immunity may have benefited the FAdV infection. RESUMO: Adenovírus aviário (FAdV) é um importante patógeno na indústria avícola global e a etiologia da síndrome da hepatite por corpúsculo de inclusão-hidropericárdio (SHH) em galinhas. Desde a década de 1990, vários surtos de SHH foram descritos em todas as áreas de produção de aves, incluindo na América do Sul, e a coinfecção entre FAdV e vírus da anemia das galinhas (CAV) pode ser agravante para todos os aspectos da SHH. Objetiva-se descrever um surto de SHH em matrizes de frangos corte, caracterizar a estirpe de FAdV envolvida e destacar a coinfecção com CAV. Foram avaliados machos reprodutores de corte com três semanas de idade, com medula óssea pálida, fígado aumentado e amarelado e esplenomegalia, timo atrofiado e um com hidropericárdio. Fígado e timo foram coletados para isolamento do vírus em embriões de galinhas SPF, PCR e histopatologia. Todas as aves acometidas e embriões inoculados foram positivos para FAdV e CAV. Os embriões inoculados tiveram óbito de 30% em até sete dias após a inoculação e alterações hepáticas por fígados esverdeados e aumentados. A análise filogenética de FAdV com base em parte da sequência do gene que codifica a proteína hexon revelou identidade com a espécie E, que se tornou disseminada em vários países. A coinfecção de FAdV e CAV resulta em maior intensidade de lesões, maior morbidade e mortalidade e em reprodutores tem alta relevância epidemiológica, em razão da transmissão vertical de ambos e da ampla distribuição geográfica das progênies infectadas. Na última década, ocorreu um aumento mundial na ocorrência de SHH em frangos de corte relacionado a falhas em biosseguridade, especialmente em reprodutores, condição que pode ter ocorrido neste episódio. A presença de FAdV e CAV em reprodutores é motivo para preocupação por reflexos negativos à imunidade e viabilidade das progênies.

The virus responsible for an outbreak of infectious laryngotracheitis (ILT) in a multi-age flock of egg layer chickens under quarantine in Brazil was characterized. Layer chickens from this area with circulating gallid herpesvirus 1 (GaHV 1) were evaluated using histopathology and molecular characterization techniques based on sequences of infected-cell polypeptide 4 (ICP4) and thymidine kinase (TK) genes. The infected chickens that were analyzed were PCR-positive for GaHV-1 in the trachea and negative in most trigeminal ganglia. The lack of ILT lesions in the conjunctiva and respiratory tissues, combined with detection of viral DNA in the trachea, was found to be associated with latent infection. The sequences from five farms obtained in the present study were identical, and there were no deletions within the 272- to 283-bp region of the ICP4 gene, as observed in the sequences of vaccine strains (CEO and TCO). The lack of a deletion in the ICP4 fragment analyzed in this study indicates that the chickens were infected with a field virus. The absence of the T252M mutation in a fragment of the TK gene, in addition to the low mortality rate observed, suggests that the outbreak in the state of Minas Gerais was not caused by a highly virulent strain but rather by a field virus of lower virulence. In addition, using phylogenetic reconstructions, it was found that this field strain was grouped together in a separate branch, apart from the previously characterized Brazilian strains. The introduction of vectored vaccines apparently has been effective in reducing clinical disease and lesions, and preventing new outbreaks of disease.

Streptococcus pneumoniae and influenza A virus (IAV) are significant agents of pneumonia cases and severe respiratory infections globally. Secondary bacterial infections, particularly by Streptococcus pneumoniae , are common in IAV-infected individuals, leading to critical outcomes. Despite reducing mortality, pneumococcal vaccines have high production costs and are serotype specific. The emergence of new circulating serotypes has led to the search for new prevention strategies that provide a broad spectrum of protection. In this context, vaccination using antigens present in all serotypes, such as Pneumococcal Surface Protein A (PspA), can offer broad coverage regardless of serotype. Employing the reverse genetics technique, our research group developed a recombinant influenza A H1N1 virus that expresses PspA (Flu-PspA), through the replacement of neuraminidase by PspA. This virus was evaluated as a bivalent vaccine against infections caused by influenza A and S. pneumoniae in mice. Initially, we evaluated the Flu-PspA virus’s ability to infect cells and express PspA in vitro, its capacity to multiply in embryonated chicken eggs, and its safety when inoculated in mice. Subsequently, the protective effect against influenza A and Streptococcus pneumoniae lethal challenge infections in mice was assessed using different immunization protocols. Analysis of the production of antibodies against PspA4 protein and influenza, and the binding capacity of anti-PspA4 antibodies/complement deposition to different strains of S. pneumoniae were also evaluated. Our results demonstrate that the Flu-PspA virus vaccine efficiently induces PspA protein expression in vitro, and that it was able to multiply in embryonated chicken eggs even without exogenous neuraminidase. The Flu-PspA-based bivalent vaccine was demonstrated to be safe, stimulated high titers of anti-PspA and anti-influenza antibodies, and protected mice against homosubtypic and heterosubtypic influenza A and S. pneumoniae challenge. Moreover, an efficient binding of antibodies and complement deposition on the surface of pneumococcal strains ascribes the broad-spectrum vaccine response in vivo. In summary, this innovative approach holds promise for developing a dual-protective vaccine against two major respiratory pathogens.

Infectious diseases can cause deleterious effects on bird species, leading to population decline and extinction. Haemosporidia can be recognized by their negative effects on host fitness, including reproductive success and immune responses. In captivity, outbreaks of haemosporidian infection have been observed in birds in zoos and aviaries. The endemic Brazilian Atlantic rainforest species Aburria jacutinga is one of the most endangered species in the Cracidae family, and wild populations of this species are currently found mainly in conservation areas in only two Brazilian states. In this study, we aimed to evaluate the effects of avian haemosporidia on hematological and biochemical parameters in two captive populations of A. jacutinga. Forty-two animals were assessed, and the haemosporidian prevalence was similar for males and females. The occurrence of haemosporidian infection in captive A. jacutinga observed in this study was similar to results found in other captive and wild birds in Brazil. We found three different lineages of haemosporidia. Two lineages were identified as Plasmodium sp., one of which was previously detected in Europe and Asia, and the other is a new lineage closely related to P. gallinaceum. A new third lineage was identified as Haemoproteus sp. We found no significant differences in hematological and biochemical values between infected and non-infected birds, and the haemosporidian lineage did not seem to have an impact on the clinical and physiological parameters of A. jacutinga. This is the first report on an evaluation of natural haemosporidian infections diagnosed by microscopic and molecular methods in A. jacutinga by hematology, blood biochemistry, and serum protein values. Determining physiological parameters, occurrence and an estimation of the impact of haemosporidia in endangered avian species may contribute to the management of species rehabilitation and conservation.

Genetic profiles of Salmonella Minnesota isolates were analyzed using pulsed-field gel electrophoresis (PFGE). In total, 13 isolates obtained from the broiler industry collected in the states of Minas Gerais (11) and São Paulo (2), as well as five recovered from cases of foodborne infections in humans in the states of Minas Gerais (2), Santa Catarina (1), and Rio Grande do Sul (2), were submitted to PFGE. These 18 S. Minnesota isolates together with other 12 of poultry origin were also subjected to antimicrobial susceptibility testing. The PFGE analysis of 18 strains of S. Minnesota generated a dendrogram that grouped the isolates with 83–90% similarity into four main clusters. Among them, cluster “A” grouped the majority of isolates (13), including two of human origin that showed 90% similarity with a broiler isolate, both recovered in Minas Gerais. The S. Minnesota isolates showed resistance to tetracycline (80%), cefoxitin (80%), ceftazidime (46.7%), nalidixic acid (23.3%), ciprofloxacin (13.3%), and streptomycin (10%). No resistance to gentamicin, chloramphenicol, meropenem, nitrofurantoin, and sulfamethoxazole-trimethoprim was found. Moreover, 23.3% of the evaluated isolates presented multi-resistance profile, all from Minas Gerais. The results highlight the importance of further studies involving S. Minnesota, which is prevalent in the Brazilian broiler flocks and could provoke foodborne infection in humans.

Ornithonyssus sylviarum is a hematophagous mite present in wild, domestic, and synanthropic birds. However, this mite can affect several vertebrate hosts, including humans, leading to dermatitis, pruritus, allergic reactions, and papular skin lesions. This study evaluated the epidemiological characteristics of O. sylviarum attacks on poultry workers, including data on laying hens, infrastructure and management of hen houses, and reports of attacks by hematophagous mites. In addition, a case of mite attack on a farm worker on a laying farm in the Midwest region in Minas Gerais is presented. It was found that 60.7% farm workers reported attacks by hematophagous mites. Correspondence analysis showed an association between reports of mite attacks in humans with (1) presence of O. sylviarum in the hen house, (2) manual removal of manure by employees, and (3) history of acaricide use. The specimens collected from the location were confirmatively identified as O. sylviarum. O. sylviarum attacks ave not been reported when manure was removed in a shorter time interval and did not use acaricide in the sheds when the removal was done by manure conveyor belt. Parasitism by O. sylviarum should be considered a relevant occupational hazard affecting employees working in direct contact with commercial egg-laying hens in Minas Gerais. We emphasized the need to monitor hen infestations by this mite to improve the development of mite control strategies. RESUMO: Ornithonyssus sylviarum é um ácaro hematófago de aves silvestres, domésticas e sinantrópicas. No entanto, este ácaro pode afetar vários hospedeiros vertebrados, incluindo humanos, levando a dermatites, prurido, reações alérgicas e lesões cutâneas papulosas. O presente estudo avaliou os aspectos epidemiológicos do ataque de O. sylviarum em trabalhadores de granjas avícolas e descreveu um relato de caso em uma granja de postura. Este estudo utilizou um banco de dados secundário com informações sobre as poedeiras, infraestrutura e manejo dos galinheiros e relatos de ataques por ácaros hematófagos. Foi apresentado um caso de ataque de ácaro a um trabalhador rural no Centro-Oeste em Minas Gerais. Houve uma frequência de 60,7% de trabalhadores que relataram ter sido atacados por ácaros hematófagos em fazendas. A análise de correspondência mostrou uma associação entre relatos de ataques de ácaros em humanos com (1) presença de O. sylviarum na fazenda, (2) remoção manual de esterco por funcionários da fazenda e (3) histórico de uso de acaricidas. Os espécimes coletados do local foram identificados como O. sylviarum. Não há ataques de O. sylviarum, quando a remoçãodo esterco é feita em menor intervalo de tempo, e não se utiliza acaricida nos galpões quando a remoção é feita por esteira transportadora de esterco. Conclui-se que o parasitismo por O. Sylviarum deve ser considerado como um risco ocupacional relevante, que afeta trabalhadores que trabalham diretamente com poedeiras comerciais em Minas Gerais. Ressaltamos a necessidade de monitorar as infestações de aves por esse ácaro, a fim de melhorar o desenvolvimento de estratégias de controle.

An outbreak of proventricular dilatation disease (PDD), a fatal inflammatory disease of psittacines (Aves: Psittaciformes), is described in native Brazilian psittacines. Twenty captive psittacines that died of suspected PDD were necropsied and 10 were submitted to histopathology, reverse transcriptase PCR (RT-PCR), and immunohistochemistry (IHC) for avian bornavirus (ABV). Examined species were one pileated parrot (Pionopsitta pileata), three vinaceous-breasted parrots (Amazona vinacea), two blue-winged macaws (Primolius maracana), one scarlet macaw (Ara macao), one chestnut-fronted macaw (Ara severa), one scaly-headed parrot (Pionus maximiliani), and one red-browed Amazon parrot (Amazona rhodocorytha). Gross examination and histopathology revealed typical PDD lesions in all birds. The presence of ABV was confirmed in four psittacines including one red-browed Amazon parrot, one blue-winged macaw, one scarlet macaw, and one chestnut-fronted macaw. In the red-browed Amazon parrot and in one blue-winged macaw, IHC demonstrated ABV antigens in the nucleus and cytoplasm of cells in various organs. This is the first description of PDD by ABV in Brazilian psittacines and indicates the necessity for adopting a strategic control plan for reducing its impact in native birds. Reporte de Caso—Enfermedad de dilatación proventricular fatal en psitácidos nativos en cautiverio en Brasil. Un brote de la enfermedad de dilatación proventricular (con las siglas en inglés PDD), que es una enfermedad inflamatoria fatal de los psitácidos (Aves : Psittaciformes ), se describe en psitácidos nativos de Brasil. Se practicó la necropsia de veinte psitácidos cautivos que se sospechó murieron de la enfermedad de dilatación proventricular y diez fueron sometidos a estudio histopatológico, por transcripción reversa y PCR, inmunohistoquímica (IHC) para bornavirus aviar (ABV). Las especies examinadas fueron uno lorito carirrojo (Pionopsitta pileata), tres loros vinosos (Amazona vinacea), dos guacamayos maracaná (Primolius maracana), un guacamayo macao (Ara macao), un guacamayo severo (Ara severa), un loro de Maximilian (Pionus maximiliani), y una amazona de frente roja (Amazona rhodocorytha). El examen macroscópico y la histopatología revelaron lesiones típicas la enfermedad de la dilatación proventricular en todas las aves. La presencia del bornavirus aviar se confirmó en cuatro psitácidos como en la amazona de frente roja, un guacamayo maracaná, el guacamayo macao y el guacamayo severo. En la amazona de frente roja y en un guacamayo maracaná, la inmunohistoquímica demostró antígenos del bornavirus aviar en el núcleo y el citoplasma de las células en diversos órganos. Esta es la primera descripción de la enfermedad de dilatación proventricular por bornavirus aviar en psitácidos brasileños y también indica la necesidad de adoptar un plan de control estratégico para la reducción de su impacto en las aves nativas.

Infection of maternal, antibody-negative chickens with chicken infectious anemia virus (CIAV) can cause clinical disease, while infection after maternal antibodies wane often results in subclinical infection and immunosuppression. Currently, vaccines are not available for vaccination in ovo or in newly hatched chickens. Development of CIAV vaccines for in ovo use depends on the ability to generate vaccines that do not cause lesions in newly hatched chicks and that can induce an immune response regardless of maternal immunity. Immune complex (IC) vaccines have been successfully used for control of infectious bursal disease, and we used a similar approach to determine if an IC vaccine is feasible for CIAV. Immune complexes were prepared that consisted of 0.1 ml containing 105.4 tissue culture infective dose 50% of CIA-1 and 0.1 ml containing 10 to 160 neutralizing units (IC Positive [ICP]10 to ICP160), in which one neutralizing unit is the reciprocal of the serum dilution required to protect 50% of CU147 cells from the cytopathic effects caused by CIA-1. Virus replication was delayed comparing ICP80 and ICP160 with combinations using negative serum (IC Negative [ICN]80 or ICN160). In addition, the number of birds with hematocrit values <28% were decreased with ICP80 or ICP160 compared to ICN80 or ICN160. Seroconversion was delayed in ICP80 and ICP160 groups. To determine if ICP80 or ICN160 protected against challenge, we vaccinated maternal, antibody-free birds at 1 day of age and challenged at 2 wk or 3 wk of age with the 01-4201 strain. Both ICP80 and ICP160 protected against replication of the challenge virus, which was measured using differential quantitative PCR with primers distinguishing between the two isolates. Thus, in principle, immune complex vaccines may offer a method to protect newly hatched chicks against challenge with field virus. However, additional studies using maternal, antibody-positive chicks in combination with in ovo vaccination will be needed to determine if immune complex vaccines will be useful to protect commercial chickens.