Trypanosoma evansi in horses from Colombia - associated infection factors

Background: Trypanosoma evansi is the most common protozoan in tropical and subtropical regions of the world, due to its ability to maintain and be transmitted by vectors such as Stomoxys spp. and Tabanus spp. This protozoan causes high morbidity and mortality rates in horses in African, American and Asian countries. In the years 2021 and 2022, a high mortality rate was reported among horses with symptoms associated with Trypanosoma spp. in the municipality of Arauca, department of Arauca, Colombia. The investigation described here was therefore carried out, seeking to identify the pathogens and risk factors that led to the death of the horses in this region of Colombia. Cases: Blood samples were collected from Colombian criollo horses and dogs, as were samples of ticks, flies and horseflies that infested the horses. A variety of tissue samples were removed from the horses a few min after their death for histopathological analysis. Two questionnaires were applied to obtain information about the horses and the environment in which they live. The results of the clinical examination revealed pale mucous membranes, jaundice, high fever, dehydration and lethargy. The horses were also infested with Amblyomma mixtum (17.6%) and Dermacentor nitens (82.4%) ticks, and with Tabanus pungens (74%), Tabanus spp. (26%), and Stomoxys calcitrans flies (100%), while the dogs were infested with Rhipicephalus sanguineus s.l. (77.7%) and Amblyomma mixtum (22.2%) ticks. The blood smear test results revealed the presence of Trypanosoma spp. in 66.6% (n = 4) of the horse blood samples, and in 50% (n = 1) of the dog blood samples. PCR performed to identify the Trypanosoma species confirmed the presence of T. evansi. Histological examination of the spleen revealed the involvement and dissemination of T. evansi in the tissues. The horses also showed the presence of Equine Infectious Anemia Virus (EIAV). Discussion: This is the first updated specific report of T. evansi in criollo horses in the savannah flood zone of the municipality of Arauca, Colombia. The main risk associated with T. evansi infection in horses was found to be infestation with the natural vector T. pungens and the mechanical vector S. calcitrans, which are efficient ectoparasites for the transmission of this parasite. The presence of T. evansi in dogs represents a constant risk to horses, because dogs may serve as a reservoir for the maintenance of the hemoparasite in the population under study. Another risk factor for horses could be the presence of vampire bats (Desmodus rotundus), a species of bat that has been described as a vector and reservoir of T. evansi in Colombia. The presence of EIAV antibodies in the horses under study can be attributed to the exposure of sick horses to vectors of this virus, such as Tabanus spp., S. calcitrans and inanimate needle-shaped fomites. This is the first study that identifies the coinfection of T. evansi and EIAV in horses in the floodplain region of Colombia. In view of the importance of these 2 pathologies to the health of horses, a greater number of tests and a larger animal population will be required to determine if this coinfection is the cause of the death of criollo horses in this region of Colombia. Lastly, the owners reported that pharmacological control with trypanocides has not been successful in most of the outbreaks that occurred during the years 2021 and 2022. This may suggest that Trypanosoma evansi is developing resistance to these drugs; therefore, specific studies will be required in the future to test this hypothesis.

Bovine viral diarrhea virus infection in cattle - antioxidant status and some biochemical parameters

Background: Bovine viral diarrhea virus (BVDV) infections in cattle result in significant economic losses due to reproductive performance deficiencies caused by gastrointestinal, respiratory system infections, and transplacental infections. BVDV is one of the most important and widespread pathogens in cattle worldwide, including Turkey. Methods such as virus neutralization, enzyme-linked immunosorbent assay (ELISA), reverse transcriptase and polymerase chain reaction (RT-PCR) are used for the detection of the disease. The diagnosis of the disease in its subclinical form is challenging due to the lengthy and costly procedures involved. Investigating oxidative stress parameters in ruminants with various diseases contributes significantly to diagnosis and prognosis. This study aimed to investigate some oxidative stress and biochemical parameters in cattle infected with BVDV. Materials, Methods & Results: In the study, blood samples were collected from 80 Simmental breed cows aged between approximately 4 and 8 years to determine the presence of BVDV antibodies using the ELISA method. Based on the results obtained, study groups were organized. The study included a group of 10 animals with positive antibody levels as the infected group, and a group of 10 animals with negative antibody levels as the healthy group. Blood samples were taken from the animals, and serum separation was ensured. In the obtained serum samples, levels of vitamin E, vitamin A, ß-Carotene, catalase, GSH-Px, and MDA were determined using spectrophotometric methods. In addition, serum total protein, albumin, alkaline phosphatase (ALP), aspartate aminotransferase (AST), glucose, low-density lipoprotein (LDL), high-density lipoprotein (HDL), calcium (Ca), and phosphorus (P) were measured using commercial test kits and an autoanalyzer. In the study, it was observed that the differences in serum MDA, vitamin E, vitamin A, ß-carotene, and catalase levels were statistically significant between the healthy and BVDV-infected groups (P < 0.001). The activity of GSH-Px was also found to be statistically different between the groups (P < 0.01). Among the biochemical parameters, HDL, LDL, and AST levels were found to be statistically significant between the healthy and BVDV-infected groups (P < 0.001). Additionally, ALP and glucose levels were found to be statistically significant (P < 0.01). However, although there were differences in the levels of total protein, albumin, Ca, and P between the groups, these results were not statistically significant. Discussion: Although the diagnosis of the disease was partially made based on clinical observations in BVDV infections, the ELISA method was used for accurate diagnosis. Furthermore, it was found that there was a significant difference in MDA concentration between the healthy and infected groups, indicating oxidative damage caused by the virus. Similarly, significant differences in vitamin E, vitamin A, ß-carotene, GSH-Px, and catalase levels were observed between the groups, indicating a decrease in antioxidant values due to the infection. In addition, differences in ALP, AST, glucose, LDL, and HDL levels were found between the groups. This difference is thought to be related to the effects of the disease agent on the liver and systemically. This study demonstrates that, in addition to the viral pathogen, antioxidant and biochemical values are important criteria in the detection of the disease.

Searching bovine papillomavirus presence in lesions seen on teats of cows

The samples were taken from 106 cows with various-looking lesions on their teats and ranged in age from 2 to 8 years. Enzyme-linked immunosorbent assay (ELISA) antigen (Ag) positive for the bovine papillomavirus (BPV) was found in 59 (55.7%) blood serum samples. PCR using FAP59/64 primers was positive for 24 (22.6%) samples. BPV-2 (40, 37.7%), BPV-6 (28, 26.4%), BPV-8 (30, 28.3%), BPV-9 (36, 34%), BPV-10 (32, 30.3%), and BPV-12 (22, 20.8%) were found in a PCR type-specific analysis of single and mixed type teat warts. The highest positivity was observed in BPV-2, BPV-9 and BPV-10 in flat and round forms, BPV-6, BPV-10, BPV-12, and mixed types in rice grain-cauliflower forms, BPV-9 and mixed types in filiform in the distribution of types based on the macroscopic appearance of teat lesions. As for the distribution of BPV types according to age, the most BPV-2 types were found in the age group of two years, the most BPV-10 types in the age group of three years, the most BPV-9 types in the age group of four years, the most BPV-8+BPV-12 types in the age group of five years, and the most mixed types between the ages of six and eight years. The existence of the virus was then checked using electron microscopy on the chosen samples (at least one investigation was conducted), and it was positively identified using BPV type-specific primers. The authors concluded that BPV detection using an ELISA (Ag) test from blood serum samples was shown to be less sensitive than BPV type-specific PCR from wart samples.

Fowl Aviadenovirus E associated with hepatitis-hydropericardium syndrome in broiler breeders / Síndrome da hepatite-hidropericárdio por Aviadenovirus e em reprodutores de corte

ABSTRACT: 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.

Fowl Aviadenovirus E associated with hepatitis-hydropericardium syndrome in broiler breeders / Síndrome da hepatite-hidropericárdio por Aviadenovirus e em reprodutores de corte

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.

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.

Biological Control in Brazil: state of art and perspectives

Brazil is one of the most extensive users of Biological Control (BC) (including both micro and macroorganisms) in the struggle to control agricultural pests. Nowadays, BC is seen not only as a measure supported by bioagents but also as an approach involving biofertilizers and biostimulants. This is part of a broader view that encompasses bioinputs, defined as agro-industrial products or processes developed from enzymes, extracts (from plants or microorganisms), microorganisms, macroorganisms (invertebrates), secondary metabolites, and semiochemicals (e.g., pheromones), all designated for BC. Sustainability is the worldwide need of the moment. Achieving this goal is a genuine concern of sustainable agriculture projects which is made possible by, among other measures, using biological products to control pests. As a leader in tropical agriculture, Brazil must increasingly use BC to meet the requirements of the international markets for agricultural products free from chemical residues. Entomological pest control involves both micro (fungi, bacteria, virus, nematodes, etc.) and macroorganisms (insects and mites). Although categorized as semiochemicals, pheromones are commonly included in BC, in this case, to monitor or control pests (e.g., the male confusion technique). In the current philosophy of Integrated Pest Management (IPM), aiming toward sustainability, BC should not be considered in isolation but as a component of IPM, i.e., as a method, similar to other control methods, to maintain pest populations below the economic injury level, which considers economic, ecological, and social criteria.(AU)

Detection of microorganisms with antibacterial activities from different industrial wastes and GC-MS analysis of crude microbial extract / Detecção de microrganismos com atividades antibacterianas de diferentes resíduos industriais e análise por GC-MS do extrato microbiano bruto

Abstract Many soil microorganisms' i.e., bacteria and fungi produce secondary metabolites called antibiotics. These are used for the treatment of some of the bacterial, fungal and protozoal diseases of humans. There is a need for isolation of a broad spectrum of antibiotics from microorganisms due to the emergence of antibiotic resistance. In the present study two antibiotic producing bacteria Klebsiella pneumoniae and Bacillus cereus were isolated from pharmaceutical and poultry feed industry of Hattar, Haripur Pakistan. Total 10 waste samples were collected from different industries (Marble, Ghee, Soap, Mineral, Steel, Poultry Feed, Pharmaceutical, Qarshi, Cosmetic and Glass). Thirty-three bacterial strains were isolated from industrial wastes of these ten different industries. Fourteen out of thirty-three bacterial strains exhibited antimicrobial activities against at least one of the test microbes considered in this study including Escherchia coli, Staphylococcus aureus and Salmonella typhi. The bacteria were isolated by standard serial dilution spread plate technique. Morphological characterization of the isolates was done by Gram staining. Nine bacterial isolates out of fourteen were initially identified as B. cereus and five as K. pneumoniae through biochemical characterization. The antibacterial activities were tested by well diffusion method. Maximum number of antibiotic producing bacteria were isolated from pharmaceutical and poultry feed industry based on the results of primary screening, the most potential isolates S9, S19, S20, S22 and S23 were selected for secondary screening. The maximum activity against E. coli and S. aureus was recorded by bacterial isolate S19 i.e zones of inhibition of 6.5mm and 9mm while S20 showed 7.5mm and 6mm zones respectively. Molecular identification was carried out on the basis of 16S rRNA sequence analysis. Finally, the isolates were identified as B. cereus accession number LC538271and K. pneumoniae accession number MT078679. Analysis of bacterial extract S20 through GC-MS indicated the presence of 8 compounds of diverse nature and structure. Present study suggests that wastes of pharmaceutical and poultry feed industry may have antibiotic producing bacteria. These bacteria could be utilized for the production of antibiotics. B. cereus and K. pneumoniae isolated from wastes of poultry feed and pharmaceutical industries have the potential to produce antibiotics and could be used to control the microbial growth.

Resumo Muitos microrganismos do solo, ou seja, bactérias e fungos produzem metabólitos secundários chamados antibióticos. Eles são usados ​​para tratamento de algumas doenças bacterianas, fúngicas e protozoárias em humanos. Há necessidade de isolamento de um amplo espectro de antibióticos de microrganismos devido ao surgimento de resistência aos antibióticos. No presente estudo, duas bactérias produtoras de antibióticos, Klebsiella pneumoniae e Bacillus cereus, foram isoladas da indústria farmacêutica e de ração avícola de Hattar, Haripur, Paquistão. Um total de 10 amostras de resíduos foi coletado de diferentes indústrias (mármore, ghee, sabão, mineral, aço, ração para aves, farmacêutica, Qarshi, cosmética e vidro). Trinta e três cepas bacterianas foram isoladas de resíduos industriais dessas dez diferentes indústrias. Quatorze das 33 cepas bacterianas exibiram atividades antimicrobianas contra pelo menos um dos micróbios de teste considerados neste estudo, incluindo Escherchia coli, Staphylococcus aureus e Salmonella typhi. As bactérias foram isoladas pela técnica de placa de diluição em série padrão. A caracterização morfológica dos isolados foi feita por coloração de gram. Nove isolados bacterianos de 14 foram inicialmente identificados como B. cereus e cinco como K. pneumoniae por meio de caracterização bioquímica. As atividades antibacterianas foram testadas pelo método de difusão em poço. O número máximo de bactérias produtoras de antibióticos foi isolado da indústria farmacêutica e de ração avícola com base nos resultados da triagem primária, os isolados mais potenciais S9, S19, S20, S22 e S23 foram selecionados para a triagem secundária. A atividade máxima contra E. coli e S. aureus foi registrada pelo isolado bacteriano S19, ou seja, zonas de inibição de 6,5 mm e 9 mm, enquanto S20 mostrou zonas de 7,5 mm e 6 mm, respectivamente. A identificação molecular foi realizada com base na análise da sequência 16S rRNA. Finalmente, os isolados foram identificados como B. cereus número de acesso LC538271 e K. pneumoniae número de acesso MT078679. A análise do extrato bacteriano S20 por meio de GC-MS indicou a presença de oito compostos de natureza e estrutura diversas. O presente estudo sugere que resíduos da indústria farmacêutica e de ração para aves podem conter bactérias produtoras de antibióticos. Essas bactérias podem ser utilizadas para a produção de antibióticos B. cereus e K. pneumoniae isolados de resíduos de rações de aves e indústrias farmacêuticas têm potencial para produzir antibióticos e podem ser usados ​​para controlar o crescimento microbiano.

Infecção pelo vírus da Febre do Nilo Ocidental em equinos no Estado de São Paulo / West Nile Fever virus infection in horses in São Paulo state, Brazil

Background: The West Nile virus (WNV) antibodies were reported in Brazil in the serum samples taken from horses andbirds in the Midwest region and Paraíba state in 2008 and 2013, respectively. In 2014, the first human case was confirmedin a rural worker in the state of Piauí and, in 2018, the virus was isolated from the central nervous system of a horse withnervous symptoms in the state of Espírito Santo. The virus is a member of the Flaviviridae family of the genus Flavivirus(neurotropic), infecting several mammalian species, with humans and horses being the most susceptible. Approximately35% of horses develop clinical signs, thus they are considered the best sentinels for this disease. The aim of this case reportis to describe the first confirmed cases of West Nile Fever (WNF) in two horses in the state of São Paulo.Cases: Two horses with neurological symptoms were treated at the Veterinary Hospital of Cruzeiro do Sul University (SãoPaulo, SP), in 2019. Both horses came from neighboring regions that have a large Atlantic Forest preservation area and arealso routes for migratory birds, known to be competent hosts for transmitting the West Nile Fever virus, such as the swallow,the falcon and the hawk. The first one had symptoms, such as weakness and sporadic seizures; however, after recovering,it was hospitalized a few days later due to kidney failure and laminitis. The second one showed incoordination, pelviclimb weakness, and was walking in circles, evolving to seizures. Both animals were euthanized, and their central nervoussystem samples and total blood samples were tested for rabies, herpes virus, and WNV; the first 2 tests showed negativeresults. Ribonucleic acids (RNA) were extracted from erythrocytes using the polymerase...

Infecção pelo vírus da Febre do Nilo Ocidental em equinos no Estado de São Paulo / West Nile Fever virus infection in horses in São Paulo state, Brazil

Background: The West Nile virus (WNV) antibodies were reported in Brazil in the serum samples taken from horses andbirds in the Midwest region and Paraíba state in 2008 and 2013, respectively. In 2014, the first human case was confirmedin a rural worker in the state of Piauí and, in 2018, the virus was isolated from the central nervous system of a horse withnervous symptoms in the state of Espírito Santo. The virus is a member of the Flaviviridae family of the genus Flavivirus(neurotropic), infecting several mammalian species, with humans and horses being the most susceptible. Approximately35% of horses develop clinical signs, thus they are considered the best sentinels for this disease. The aim of this case reportis to describe the first confirmed cases of West Nile Fever (WNF) in two horses in the state of São Paulo.Cases: Two horses with neurological symptoms were treated at the Veterinary Hospital of Cruzeiro do Sul University (SãoPaulo, SP), in 2019. Both horses came from neighboring regions that have a large Atlantic Forest preservation area and arealso routes for migratory birds, known to be competent hosts for transmitting the West Nile Fever virus, such as the swallow,the falcon and the hawk. The first one had symptoms, such as weakness and sporadic seizures; however, after recovering,it was hospitalized a few days later due to kidney failure and laminitis. The second one showed incoordination, pelviclimb weakness, and was walking in circles, evolving to seizures. Both animals were euthanized, and their central nervoussystem samples and total blood samples were tested for rabies, herpes virus, and WNV; the first 2 tests showed negativeresults. Ribonucleic acids (RNA) were extracted from erythrocytes using the polymerase...(AU)

Retrospective study of poxviruses diagnosed in cattle from Goiás State, Brazil (2010-2018) / Estudo retrospectivo de poxviroses diagnosticadas em bovinos no estado de Goiás (2010-2018)

A retrospective study of poxvirus infections diagnosed in cattle from Goiás state (GO), Brazil, from 2010 to 2018, was performed. All cases have been investigated by the GO Official Veterinary Service (Agrodefesa), from which technical forms and protocols of veterinary diagnosis laboratories were reviewed. In most cases, samples of oral or cutaneous tissues and/or swabs were submitted for virological diagnosis by polymerase chain reaction (PCR) and/or virus isolation. Thirty seven outbreaks/cases of vesicular disease were notified in cattle of 25 counties; in 33 cases the animals presented lesions clinically compatible with poxviruses. The etiology of 25 out of 33 outbreaks/cases was confirmed as poxviruses by PCR and/or viral isolation: 13 as bovine vaccinia virus (VACV), six as pseudocowpox virus (PCPV), five as bovine papular stomatitis virus (BPSV) and one coinfection (VACV and an Orf virus-like parapoxvirus). The laboratory confirmed that cases occurred mainly in dairy cattle (19/25) and during the dry season (22/25). In adult cattle, gross changes were observed mainly in the teats and udder and included vesicles, ulcers, crusts, papules and scars and varied of type, severity and affected region, depending on the poxvirus species. In calves, the main lesions were ulcers in the mouth and muzzle. Zoonotic lesions compatible with poxvirus infections were observed for all diagnosed poxviruses, affecting especially the hands of milkers and other farm workers. Our data demonstrate the sanitary and economic relevance of these diseases and the wide circulation of different poxviruses in cattle from GO.

Foi realizado um estudo retrospectivo das infecções por poxvírus diagnosticadas em bovinos do estado de Goiás (GO), entre 2010 e 2018. Todos os casos foram investigados pela Agência Goiana de Defesa Agropecuária (Agrodefesa). Foram revisados formulários técnicos e protocolos de laboratórios de diagnóstico veterinário. Na maioria dos casos, amostras de tecidos orais ou cutâneos e/ou swabs foram encaminhadas para diagnóstico virológico. Foram notificados 37 surtos/casos de doença vesicular em bovinos em 25 municípios; em 33 casos os animais apresentavam lesões clinicamente compatíveis com poxvírus. A etiologia de 25 de 33 surtos/casos foi confirmada como poxvírus por PCR e/ou isolamento viral: 13 como vírus vaccínia (VACV), seis como vírus pseudocowpox (PCPV), cinco como vírus da estomatite papular bovina (BPSV) e um caso de coinfecção (VACV e um parapoxvírus semelhante ao Orf vírus). Os casos confirmados laboratorialmente ocorreram principalmente em bovinos leiteiros (19/25) e durante a estação seca (22/25). Em bovinos adultos, alterações macroscópicas foram observadas principalmente nas tetas e úbere e incluíram vesículas, úlceras, crostas, pápulas e cicatrizes e variaram quanto ao tipo, gravidade e região afetada, dependendo da espécie do poxvírus. Em bezerros, as principais lesões foram úlceras na boca e focinho. Lesões zoonóticas compatíveis com infecção por poxvírus foram observadas em todas as poxviroses diagnosticadas, afetando principalmente as mãos dos ordenhadores e outros trabalhadores rurais. Nossos dados demonstram a relevância sanitária e econômica dessas doenças e a ampla circulação de diferentes poxvírus em bovinos de GO.

Retrospective study of poxviruses diagnosed in cattle from Goiás State, Brazil (2010-2018) / Estudo retrospectivo de poxviroses diagnosticadas em bovinos no estado de Goiás (2010-2018)

A retrospective study of poxvirus infections diagnosed in cattle from Goiás state (GO), Brazil, from 2010 to 2018, was performed. All cases have been investigated by the GO Official Veterinary Service (Agrodefesa), from which technical forms and protocols of veterinary diagnosis laboratories were reviewed. In most cases, samples of oral or cutaneous tissues and/or swabs were submitted for virological diagnosis by polymerase chain reaction (PCR) and/or virus isolation. Thirty seven outbreaks/cases of vesicular disease were notified in cattle of 25 counties; in 33 cases the animals presented lesions clinically compatible with poxviruses. The etiology of 25 out of 33 outbreaks/cases was confirmed as poxviruses by PCR and/or viral isolation: 13 as bovine vaccinia virus (VACV), six as pseudocowpox virus (PCPV), five as bovine papular stomatitis virus (BPSV) and one coinfection (VACV and an Orf virus-like parapoxvirus). The laboratory confirmed that cases occurred mainly in dairy cattle (19/25) and during the dry season (22/25). In adult cattle, gross changes were observed mainly in the teats and udder and included vesicles, ulcers, crusts, papules and scars and varied of type, severity and affected region, depending on the poxvirus species. In calves, the main lesions were ulcers in the mouth and muzzle. Zoonotic lesions compatible with poxvirus infections were observed for all diagnosed poxviruses, affecting especially the hands of milkers and other farm workers. Our data demonstrate the sanitary and economic relevance of these diseases and the wide circulation of different poxviruses in cattle from GO.(AU)

Foi realizado um estudo retrospectivo das infecções por poxvírus diagnosticadas em bovinos do estado de Goiás (GO), entre 2010 e 2018. Todos os casos foram investigados pela Agência Goiana de Defesa Agropecuária (Agrodefesa). Foram revisados formulários técnicos e protocolos de laboratórios de diagnóstico veterinário. Na maioria dos casos, amostras de tecidos orais ou cutâneos e/ou swabs foram encaminhadas para diagnóstico virológico. Foram notificados 37 surtos/casos de doença vesicular em bovinos em 25 municípios; em 33 casos os animais apresentavam lesões clinicamente compatíveis com poxvírus. A etiologia de 25 de 33 surtos/casos foi confirmada como poxvírus por PCR e/ou isolamento viral: 13 como vírus vaccínia (VACV), seis como vírus pseudocowpox (PCPV), cinco como vírus da estomatite papular bovina (BPSV) e um caso de coinfecção (VACV e um parapoxvírus semelhante ao Orf vírus). Os casos confirmados laboratorialmente ocorreram principalmente em bovinos leiteiros (19/25) e durante a estação seca (22/25). Em bovinos adultos, alterações macroscópicas foram observadas principalmente nas tetas e úbere e incluíram vesículas, úlceras, crostas, pápulas e cicatrizes e variaram quanto ao tipo, gravidade e região afetada, dependendo da espécie do poxvírus. Em bezerros, as principais lesões foram úlceras na boca e focinho. Lesões zoonóticas compatíveis com infecção por poxvírus foram observadas em todas as poxviroses diagnosticadas, afetando principalmente as mãos dos ordenhadores e outros trabalhadores rurais. Nossos dados demonstram a relevância sanitária e econômica dessas doenças e a ampla circulação de diferentes poxvírus em bovinos de GO.(AU)

Investigation of Chlamydia sp., Morbillivirus sp., Parvovirus sp., Leishmania sp. and Alphacoronavirus sp. in captive giant anteaters (Myrmecophaga tridactyla) / [Investigação de Chlamydia sp., Morbillivirus sp., Parvovirus sp., Leishmania sp. e Alphacoronavirus sp. em tamanduás-bandeira (Myrmecophaga tridactyla) em cativeiro]

This research aimed to investigate the occurrence of Chlamydia sp., Morbillivirus sp., Parvovirus sp., Leishmania sp. and Alphacoronavirus sp. in captive giant anteaters. Blood and fecal samples were taken from 16 animals in institutions from the states of Minas Gerais, Bahia and Distrito Federal, which had been in captivity for at least a year. A commercial rapid chromatographic immunoassay test was used for detecting coronavirus and parvovirus antigens, in addition to antibodies against leishmaniasis, all results being negative. In the case of the test for antibodies against distemper, four (4/16; 25%) anteaters had an average titration, two (2/16; 12.5%) a low titration and ten (10/16; 62.5%) were non-reactive. Using the DOT-ELISA (dot blotting) method for detection of immunoglobulin G, only one specimen obtained a 1 : 40 titration. For the polymerase chain reaction tests for Leishmania and Chlamydia, all samples were negative.

Esta pesquisa teve como objetivo investigar a ocorrência de Chlamydia sp., Morbillivirus sp., Parvovirus sp., Leishmania sp. e Alphacoronavirus sp. em tamanduás-bandeira cativos. Foram colhidas amostras de sangue e fezes de 16 animais em instituições dos estados de Minas Gerais, Bahia e Distrito Federal, que estavam em cativeiro há pelo menos um ano. Um teste comercial rápido de imunoensaio cromatográfico foi usado para detectar antígenos de coronavírus e parvovírus, além de anticorpos contra a leishmaniose, sendo todos os resultados negativos. No caso do teste para anticorpos contra a doença, quatro (4/16; 25%) tamanduás apresentaram titulação média, dois (2/16; 12,5%) uma titulação baixa e dez (10/16; 62,5%) não foram reativos. A partir do método DOT-ELISA (dot blotting) para detecção de imunoglobulina G, apenas um espécime obteve uma titulação de 1: 40. Para os testes de reação em cadeia da polimerase para Leishmania e Chlamydia, todas as amostras foram negativas.

Blue tongue virus in dairy cattle in the Southern region of Minas Gerais, Brazil - serological survey

Background: Blue tongue (BT) is a noncontagious viral disease transmitted by hematophagous arthropods, especially of the genus Culicoides. The economic impact of the disease is related not only to deaths in sheep herds but also to the possible correlation of virus infection with the development of other diseases, such as pneumonia, abortion and movement problems. The economic losses caused by Blue Tongue are linked to restrictions on the import and export of animals and their genetic material and to the reproductive disorders associated with this disease. In addition, the fact that cattle take the role of reservoir, combined with the care by other countries with outbreaks of infection and biological contamination of their products, hinders trade in Mercosul, United States and Europe. Cattle are affected by Blue Tongue Virus in endemic areas and in some epidemic areas, but the development of clinical disease is rare. The clinical signs, when evident, range from reproductive losses, such as embryonic death, abortion, fetal malformation, temporary sterility, infertility in bulls, stillbirths and the birth of weak animals. The objective of this study was to determine the epidemiological aspects of Blue Tongue Virus (BTV) infection in dairy cattle in the Lavras region, state of Minas Gerais, Brazil. Materials, Methods & Results: A cross-sectional study was conducted to evaluate the frequency of cattle and herds seropositive for Blue Tongue in the southern region of Minas Gerais. In this study, 54 dairy farms were visited. A total of 586 serum samples were collected from cows of reproductive age. Sampling was random, and serum samples were collected from lactating cows over 24 months of age by puncture of the jugular vein and/or coccidian vein. The samples were transported and stored at the Setor de Patologia Veterinária, at the Universidade Federal de Lavras (SPV-UFLA), where they were centrifuged, and the serum aliquots were obtained, transferred to microtubes and kept at -20°C until the serological tests were performed. The samples were tested with the agarose gel immunodiffusion test (AGID) for anti-blue tongue virus antibodies. The AGID test is more practical and is the main method used to identify Blue Tongue Virus seroprevalence in different ruminant species. They are considered important tools for epidemiological surveillance of the disease. A prevalence of 83.28% was observed among animals that were seropositive for Blue Tongue Virus (488/586; IC 95% = 80.0 - 86.21). In addition, 100% (54/54; IC 95% = 93.4 - 100.0) of the farms had at least 1 positive animal, with rates ranging from 45.45% to 100% within the herds and where 22.22% of the farms had rates of 100% of the animals being positive. Discussion: Blue Tongue is a disease known to affect domestic and wild ruminants in Brazil. However, there is a lack of more precise information about its epidemiology and occurrence in the country and of joint efforts of researchers, producers and the government to understand in detail both the biology of vectors and the viral biology of Blue Tongue Virus in Brazil. This is the first record of detection of anti-blue tongue virus antibodies in cattle in the southern region of Minas Gerais. The results suggest that Blue Tongue Virus is present in cattle in the study area.

Protocolo para diagnóstico molecular de imunodeficiência e leucemia em gatos domésticos / Protocol for molecular diagnosis of immunodeficiency and leukemia, in domestic cats

Diversas afecções que acometem os felinos domésticos, como as retroviroses, não possuem tratamento efetivo. Tal fato torna relevante o estudo dessas doenças, em virtude da baixa eficácia de cura e caráter majoritariamente vitalício. Entre as retroviroses que mais acometem os felinos estão a imunodeficiência felina (FIV) e a leucemia felina (FeLV). O diagnóstico é obtido pela associação do exame clínico, geralmente inconclusivo, com exames laboratoriais complementares. Testes moleculares, como a reação em cadeia da polimerase (PCR), são eficientes para a detecção do DNA proviral e podem ser utilizados na rotina diagnóstica. Diante disso, o objetivo deste estudo foi comprovar a eficiência do protocolo molecular Nested PCR para diagnosticar FIV e FeLV. Para tal, amostras de sangue e/ou medula de 41 gatos domésticos foram coletadas por meio de punção venosa ou de medula e encaminhadas ao laboratório Oncells Biotecnologia. Os seguintes pares de primers foram adotados para o Nested PCR: FF1 e FF2 para o primeiro ciclo, com um amplicon de 1325pb para FIV e 490pb para a FeLV. Para o segundo foi utilizada a combinação F14, F15, FE4 e FE7, com um amplicon de 1138pb para FIV e 306pb para FeLV. As bandas correspondentes às esperadas para FeLV foram detectadas pela observação dos géis, porém, além de outras bandas inespecíficas, não foram observadas bandas correspondentes à FIV. Os resultados confirmam a capacidade de detecção do patógeno da FeLV pela técnica empregada. No entanto, novos ajustes do protocolo são necessários.

Several affections that affect domestic cats, such as retroviruses, do not have effective treatment. This fact makes the study of these diseases relevant, due to the low healing efficacy and mostly lifelong character. Among the retroviruses that most affect felines are feline immunodeficiency (FIV) and feline leukemia (FeLV). The diagnosis is obtained by associating the clinical examination, which is generally inconclusive, with complementary laboratory tests. Molecular tests, such as the polymerase chain reaction (PCR), are efficient for proviral DNA detection and can be used in the diagnostic routine. Therefore, this study aimed to prove the efficiency of the molecular protocol Nested PCR to diagnose FIV and FeLV. For this purpose, blood and/ or bone marrow samples from 41 domestic cats were collected through venipuncture or bone marrow and sent to the Oncells Biotechnology laboratory. The following primer pairs were adopted for the Nested PCR: FF1 and FF2 for the first cycle, with an amplicon of 1325bp for FIV and 490bp for FeLV. For the second, the combination F14, F15, FE4, and FE7 was used, with an amplicon of 1138bp for FIV and 306bp for FeLV. The bands corresponding to those expected for FeLV were detected by observing the gels; however, in addition to other non-specific bands, bands corresponding to FIV were not observed. The results confirm the ability to detect the FeLV pathogen by the technique employed. Nevertheless new protocol adjustments are required.

Acute erythroid leukemia in a cat infected with feline leukemia virus

Background: Erythroid leukemia is a myeloproliferative hematopoietic disorder considered acute when there is a predominance of blasts in the bone marrow. It is frequently reported in cats infected with feline leukemia virus, but it is unclear whether this virus is involved in the oncogenesis. The clinical signs in cats are anorexia, apathy, weight loss, with evolution from 2 weeks to 2 months, pale mucous membranes, hemorrhages, ascites, salivation, and dyspnea due to pleural effusion. This affection responds little to chemotherapy with an unfavorable prognosis. The aim of this study is to report a case of a feline leukemia virus infected cat with the onset of severe hemolytic anemia. Case: A 8-year-old male mixed breed cat was attended with a history of anorexia, oligodipsia, apathy, progressive weight loss, and yellowish color of urine for 7 days. Laboratorial exams showed anemia (with metarubricytes, acanthocytes and ghost cells), leukocytosis and FeLV reagent test. The cat underwent treatment with methylprednisolone acetate and supportive care. One day later, the animal returned with icteric mucous membranes, and emesis. A blood count was performed that found worsening anemia, increased leukocytosis, and lymphocytosis. Abdominal ultrasound showed cholangiohepatitis and lymphadenomegaly in mesenteric lymph nodes. Treatment was started with ondansetron, metronidazole, and amoxicilin with potassium clavulanate. The cat returned after 3 days and laboratorial exams revealed worsening of blood parameters, so blood transfusion was performed. After 2 days, the patient started with dyspnea and hypothermia, that evolved to cardiorespiratory arrest. The body was sent to necropsy and histopathology, where blast cells and rubricytes were found in blood vessels of various organs. The bone marrow was markedly cellular with complete disappearance of adipose tissue. Most of the cells were blasts with abundant and eosinophilic cytoplasm, central nucleus with finely dotted chromatin and a large nucleolus. There were rubricytes, which made possible to confirm acute erythroid leukemia as a morphological diagnosis. Discussion: The clinical signs observed in acute erythroid leukemia are lethargy, inappetence, fever, splenomegaly, mild lymphadenomegaly, associated with leukocytosis, severe anemia, and thrombocytopenia. The reported animal presented signs similar to those described in the literature except that there was no change in platelet counts. The diagnosis of leukemia was reached after histopathology, and it is made when is observed more than 30% of myeloblasts and monoblasts together or when the blast cells count including rubriblasts is greater than 30%. Although chemotherapy, the prognosis is usually poor. It is essential to perform the myelogram for the diagnosis of myeloid leukemias in vivo. In this report, we only achieve final diagnosis after the cat's death, due to the aggressive behavior of the disease. Clinicians must be aware of the likely development of acute erythroid leukemia whenever a feline leukemia virus infected cat presents hemolytic anemia to get an early diagnosis, since this is an extremely aggressive disease, to propose prompt chemotherapy and give the patient a longer survival period.

Classification of lymphoma in cats and its relationship with the detection of feline leukemia vírus proviral DNA / Classificação do linfoma em gatos e sua relação com a detecção do DNA pró-viral do vírus da leucemia felina

In this retrospective and prospective study, histopathological and immunohistochemical analyses of 62 cases of lymphomas in cats were performed to classify the anatomic forms and subtypes, according to the WHO guidelines, and correlate it to FeLV proviral DNA detected using PCR. The most common anatomical form was gastrointestinal (40.3%, 25/62), followed by multicentric (29%, 18/62), mediastinal (17.7%, 11/62) and extranodal (12,9%, 8/62). Among the lymphoma subtypes, diffuse large B-cell lymphoma (DLBCL) (30.6%, 19/62) was the most commonly diagnosed followed by peripheral T-cell lymphoma (PTCL) (29%, 18/62) and enteropathy associated T-cell lymphoma type 2 (14.5%, 9/62). DNA extraction from paraffin-embedded neoplastic tissue was obtained in 28 cases and FeLV proviral DNA was detected by PCR, in 23 of these. Of the cases presenting with FeLV proviral DNA, nine (32%) were of the multicentric form, five (22%) of the mediastinal and extranodal forms and four (17%) of the gastrointestinal form. The most frequent subtypes with FeLV proviral DNA, independent of the anatomical form, were DLBCL (39.1%, 9/23) and PTCL (34.7%, 8/23). The presence of the FeLV proviral DNA in 23 cats of this study, probably had association with the multicentric form of lymphoma and higher occurrence in the DLBCL and PTCL subtypes.

Neste estudo retrospectivo e prospectivo, análises histopatológicas e imuno-histoquímicas de 62 casos de linfomas em gatos foram realizadas para classificar as formas anatômicas o e subtipos do linfoma, de acordo com as diretrizes da OMS. Além disso, foi realizada a extração de DNA dos tumores incluídos na parafina para obtenção de DNA pró-viral do FeLV por PCR, e relacionada com os exames anteriores. A forma anatômica mais comum foi a gastrointestinal (40.3%, 25/62), seguida pela multicêntrica (29%, 18/62), mediastinal (17,7%, 11/62) e extranodal (12,9%, 8/62). Entre os subtipos de linfoma, o linfoma difuso de grandes células B (DLBCL) (30.6%, 19/62) foi o mais comumente diagnosticado, seguido por linfoma de células T periférico (PTCL) (29%, 18/62) e o linfoma de células T associado a enteropatia tipo 2 (14.5%, 9/62). A extração de DNA de tecido neoplásico emblocado em parafina foi obtida em 28 casos e o DNA pró-viral de FeLV foi detectado por PCR, em 23 deles. Dos casos com DNA pró-viral do FeLV, nove (32%) eram da forma multicêntrica, cinco (22%) das formas mediastinal e extranodal e quatro (17%) da forma gastrointestinal. Os subtipos mais frequentes com DNA pró-viral do FeLV, independente da forma anatômica, foram DLBCL (39.1%, 9/23) e PTCL (34.7%, 8/23). A presença do DNA pró-viral do FeLV em 23 gatos deste estudo, provavelmente teve associação com a forma multicêntrica do linfoma e maior ocorrência nos subtipos DLBCL e PTCL.

Development of a sandwich enzyme-linked immunosorbent assay based on single-domain antibody for detecting goose parvovirus infection / [Desenvolvimento de um ensaio imunoadsorvente ligado a enzima sandwich baseado em anticorpo de domínio único para detecção de infecção por parvovírus de ganso]

A infecção por parvovírus de ganso (GPV) é uma doença infecciosa altamente patogênica em gansinhos e patinhos de Muscovy. Para detectar o antígeno GPV, desenvolvemos um novo ensaio imunoenzimático (ELISA) sanduíche usando um anticorpo de domínio único de cadeia pesada específico contra a proteína GPV NS1 como anticorpo de detecção. Os limites de detecção da proteína GST-NS1 e do título do vírus da cepa GPV H1 foram 5 ng/mL e 102,9 TCID50/mL, respectivamente. O ELISA sanduíche foi específico para GPV sem reatividade cruzada com outros vírus comuns de ganso, incluindo vírus Tembusu, circovírus de ganso, adenovírus de aves, vírus Newcastle ou vírus H9 da gripe aviária. Um total de 118 amostras de swab cloacal foram usadas para detectar o antígeno GPV usando o ELISA sanduíche e reação em cadeia da polimerase com uma taxa de coincidência de 91,5%. A sensibilidade e especificidade do ELISA sanduíche foram de 91,6% e 91,4%, respectivamente. Esses resultados sugerem que este ELISA sanduíche pode ser aplicado para detectar GPV.

Cinomose em canídeos silvestres no Brasil / Distemper in wild canids in Brazil

A cinomose é uma doença viral multissistêmica causada por um Morbillivirus. No Brasil, existem seis espécies de canídeos silvestres vulneráveis a essa enfermidade. Dessa forma, este trabalho tem como objetivo apresentar uma revisão sobre a situação da cinomose em canídeos silvestres no Brasil e os impactos causados na fauna brasileira. A transmissão do vírus ocorre através do contato com amostras contaminadas, a exemplo dos aerossóis, excretas e secreções de indivíduos infectados. Os sinais clínicos mais prevalentes são de ordem neurológica, entretanto, também podem ser identificadas manifestações respiratórias, cutâneas e gastrointestinais. Portanto, o diagnóstico consiste na avaliação da sintomatologia apresentada em conjunto com testes específicos, como isolamento viral, ELISA, imunofluorescência e RT-PCR. Atualmente, não existe tratamento específico. Desta forma, são realizados apenas cuidados paliativos. Os grandes carnívoros são os mamíferos mais ameaçados do mundo, sobretudo, em consequência dos impactos causados pela redução do habitat natural, associada à expansão territorial humana. As epidemias também justificam as altas taxas de mortalidades desses animais, o que pode estar relacionado com comportamentos sociais e de farejamento, assim como o crescente contato entre animais silvestres e domésticos devido à urbanização e à proximidade genética que os canídeos silvestres têm com os cães domésticos, tornando-os suscetíveis às infecções. Visto que a cinomose é uma patologia emergente em populações de carnívoros silvestres e que a presença de cães domésticos não vacinados em áreas de conservação representa um grande risco de contaminação, conclui-se que a não vacinação está diretamente associada à perpetuação do vírus no meio selvagem.

Distemper is a multisystem viral disease caused by a Morbillivirus. In Brazil, there are six species of wild canids, vulnerable to this disease. Therefore, the present work aims to develop a review on the situation of distemper in wild canids in Brazil and the impacts caused on the Brazilian fauna. The virus transmission occurs through contact with contaminated samples, such as aerosols, excreta, and secretions from infected individuals. The most prevalent clinical signs are neurological; however, respiratory, cutaneous and gastrointestinal manifestations can also be identified. Thus, the diagnosis consists of evaluating the symptoms presented together with specific tests, such as viral isolation, ELISA, immunofluorescence, and RT-PCR. Currently, there is no specific treatment. Therefore, only palliative care is performed. Large carnivores are the most threatened mammals in the world, mainly as a result of the impacts caused by the reduction of natural habitat, associated with human territorial expansion. Epidemics also justify the high mortality rates of these animals, which may be related to social and sniffing behaviors, as well as the increasing contact between wild and domestic animals due to urbanization and the genetic proximity that wild canids have with domestic dogs, making them susceptible to infections. Since distemper is an emerging pathology in populations of wild carnivores and the presence of unvaccinated domestic dogs in conservation areas represents a great risk of contamination, it is concluded that non-vaccination is directly associated with the perpetuation of the virus in the wild.

CD4+ and CD8+ T lymphocyte counts and ratio in cats with chronic gingivostomatitis and naturally infected with feline immunodeficiency virus: a preliminary study / Razão e contagem de linfócitos T CD4+ e linfócitos T CD8+ em gatos com gengivoestomatite crônica e infectados naturalmente pelo vírus da imunodeficiência felina: estudo preliminar

This study aimed to evaluate the CD4+ and CD8+ T lymphocytes counts and CD4+: CD8+ ratio in a colony of cats with chronic gingivostomatitis (CGS). We used forty domestic short-haired cats inhabiting the same colony. Ten cats with CGS were immunodeficiency virus-positive (group IV), and ten with CGS were immunodeficiency virus-negative (group III). As a control, twenty cats without CGS were used: ten cats were immunodeficiency virus-positive (group II) and ten cats were immunodeficiency virus-negative (group I). We employed flow cytometry to count CD4+ and CD8+ T lymphocytes. In cats infected with the immunodeficiency virus, the presence of CD4+ lymphocytes were lower both for animals with and without CGS. Conversely, not immunodeficiency virus-infected cats with CGS had a higher amount of CD4+ when compared to seronegative animals without CGS. The counts of CD8+ T lymphocytes showed no significant difference among cats with CGS, whether infected with immunodeficiency virus or not. The CD4+: CD8+ ratio was only different for group III, which was higher than any other group. No difference was observed for total lymphocyte number and CD8+ among groups. By contrast, mean CD4+ levels were different, with cats from groups III and IV showing higher levels than those from groups I and II. The flow cytometry could be a useful tool for the diagnosis and prognosis of cats with CGS infected by the immunodeficiency virus.

Este estudo teve como objetivo avaliar a contagem e a razão de linfócitos T CD4+ e CD8+ em uma colônia de gatos com gengivoestomatite crônica (CGS). Foram analisados quarenta gatos domésticos que habitavam a mesma colônia. Dez gatos com CGS foram positivos para o vírus da imunodeficiência (grupo IV), e dez com CGS foram negativos para o vírus da imunodeficiência (grupo III). Como controle, vinte gatos sem CGS foram usados: dez gatos foram positivos para o vírus da imunodeficiência (grupo II) e dez gatos foram negativos para o vírus da imunodeficiência (grupo I). Empregou-se a citometria de fluxo para contagem de linfócitos T CD4+ e CD8+. Nos gatos infectados pelo vírus da imunodeficiência, a presença de linfócitos CD4+ foi menor tanto para os animais com e sem CGS. Por outro lado, gatos não infectados e com CGS apresentaram maior quantidade de linfócitos CD4+ quando comparados a animais soronegativos sem CGS. A contagem de linfócito T CD8+ não mostrou diferença significativa entre gatos com CGS, infectados ou não com o vírus da imunodeficiência. A razão CD4+:CD8+ foi diferente apenas para o grupo III, que foi maior do que qualquer outro grupo. Não foi observada diferença para o número total de linfócitos e CD8+ entre os grupos. Em contraste, os níveis médios de CD4+ foram diferentes, com os gatos dos grupos III e IV apresentando níveis mais elevados do que os dos grupos I e II. A citometria de fluxo pode ser uma ferramenta útil para o diagnóstico e prognóstico de gatos com CGS infectados pelo vírus da imunodeficiência.

Marek's disease in roosters in the Potiguar semi-arid region

Background: Marek's disease (MD) is a transmissible disease in chickens caused by Gallid alphaherpesvirus 2 (GaHV-2). The infection is characterized by lymphocyte cellular infiltrates in peripheral nerves and other organs and tissues, including the skin; which can lead to dysfunction causing progressive asymmetric paresis and complete spastic paralysis of body extremities. Dermatitis and cardiac myositis caused by GaHV-2 in free-range chickens has rarely been described in Brazil. This reports the occurrence of the disease with a confirmatory molecular diagnosis in free-range poultry showing signs of dermatitis, poor performance, and cachexia and no mortality in the semi-arid Potiguar region. Cases: Twenty roosters of the Shamo lineage, among a brood of 42 birds, had a history of progressive weight loss and skin lesions. Two birds with poor body condition, erythema, and scaling of the skin in the head and cervical regions were sent for clinical care. All birds were between 12 and 18 months of age and were vaccinated against Newcastle disease and Fowlpox with only a few receiving vaccines against MD and Gumboro disease. According to the owner's report, some birds were previously kept outdoors, and when they were transferred to a small shed with little air circulation, they began to develop clinical signs after approximately 15 days. The first signs of the disease were also reported to have appeared 2.5 months before clinical care and, in the meantime, several treatments were instituted without success. Owing to the general condition of the animals and inconclusive clinical suspicion, the birds were subjected to euthanasia and necropsy. Tissue samples were collected for histopathological and polymerase chain reaction analyses to search for the GaHV-2 DNA meq gene. The main clinicopathological findings were erythema (47%, 20/42) and desquamation of skin and mild, prominent white multifocal areas in the heart. Histopathology revealed infiltration of pleomorphic lymphoblastic cells in the skin, heart, and sciatic nerve. The amplification of the L-meq and meq oncoprotein genes in these organs and in the liver, confirmed the infection by GaHV-2, consistent with that of a field strain. Discussion: MD was confirmed based on the macroscopic and histological lesions, and with the detection of GaHV-2 DNA in the affected tissues. The unusual clinical presentation represented an initial challenge for diagnosis. The clinical history was important to lead to the suspicion of MD, as roosters initiated clinical signs 15 days after they were transferred to a small shed with poor air circulation. This probably favored the high viral concentration and disease transmission among susceptible birds in the brood because the feather follicle is the primary site of viral replication for transmission; and desquamation of infected epithelial cells favor airborne horizontal transmission to susceptible chickens. The roosters had not been vaccinated against MD, which probably favored the infection, as vaccination is known to be a fundamental approach for MD control for effective growth of the poultry industry. Clinical findings and lesions, together with viral molecular detection, were fundamental for the diagnosis, a premise for the application of adequate prevention and control measures for the disease in breeding. This is the first report of MD with a confirmatory molecular diagnosis in northeastern Brazil.