New records of Amblyomma ticks parasitizing neotropical primates in Brazil.

Amblyomma is an important tick genus for animal and human health, with some species being the vectors of zoonotic pathogens, such as Rickettsia rickettsii, in the Neotropical region. Knowing their hosts may help to understand the distribution of these agents and decrease the occurrence of clinical cases. Primates are intelligent and adaptable animals that can get close to humans in the search for food. So, they may be an important epidemiological link for the spread of these ticks. Beyond that, primates also suffer from these infections, serving as sentinels for different diseases. Thus, the present study aims to report the parasitism by Amblyomma spp. on six species of Neotropical primates from different locations in Brazil. The 337 collected ticks were morphologically identified using stereomicroscopes and taxonomic keys, and six distinct species of ticks were identified. We report here the first record of nymphs of the tick species Amblyomma cajennense sensu stricto on Alouatta belzebul, a male of Amblyomma fuscum on Alouatta guariba clamitans, nymphs of Amblyomma sculptum on Leontopithecus chrysopygus and Callithrix aurita, as well as nymphs of Amblyomma geayi on Saimiri collinsi. Of the 337 tick specimens collected, 256 (75,96%) were nymphs. The importance of primates in the life cycle of these species remains to be elucidated.

Survey of euthanasia methods used in neotropical chiropterans in São Paulo State, Brazil: critical analyses and proposal for standardization of bat euthanasia methods / evantamento dos métodos de eutanásia utilizados em quirópteros neotropicais no estado de São Paulo, Brasil: análise crítica e proposta de padronização de métodos de eutanásia em morcegos

Zoonotic Surveillance Divisions (ZSDs) rescue and euthanize bats in contact with humans. The euthanasia procedure should be conducted safely for the animal and caretaker and minimal stress for the animal is important. The objectives of this study were: i. evaluate the methods of euthanasia used by ZSDs in the State of São Paulo; ii. compare the methods with national and international guidelines for animal euthanasia practices; iii. assess the methodologies considering taxonomy and eating habits of the main bat species, and iv. propose standardization of euthanasia procedures. Sixty-five ZSDs locations received an online questionnaire or were contacted by telephone and 33 ZSDs (50.8%) responded and are distributed in 11 mesoregions in the state to remain anonymous. The euthanasia methods were divided into chemical (injectable or inhalation), physical, or mixed methods. Bat specimens (n = 550) were identified and classified to evaluate the main genera found in the state. The location of the ZSD, species, eating habits, and the method of euthanasia used were analyzed. The specimens by bat families were Molossidae (n = 340), Phyllostomidae (n = 171), and Vespertilionidae (n = 39). Chemical methods were used in 25 ZSDs (75.75%), physical in 5 (15.15%), and mixed in 3 (9.09%). There is no uniformity or standardization in bat euthanasia methods used by ZSDs, although most are based on acceptable chemical methods. It was proposed an algorithm to assist the veterinarian in choosing the method of euthanasia for bats that will allow standardizing euthanasia procedures for this species, considering physiological differences, and respecting technical, bioethical, and animal welfare guidelines.(AU)

As Divisões de Vigilância de Zoonoses (DVZs) coletam e eutanasiam morcegos que tiveram contato com humanos. O procedimento de eutanásia precisa ocorrer de modo seguro para o animal e para o manipulador, sendo importante garantir o mínimo de estresse ao animal. Os objetivos deste estudo foram: i. avaliar os métodos de eutanásia praticados nas DVZ do estado de São Paulo; ii. comparar esses métodos com os guias nacionais e internacionais de práticas de eutanásia animal; iii. avaliar as metodologias aplicadas considerando a taxonomia e os hábitos alimentares das principais espécies de ocorrência de morcegos; e iv. propor a padronização dos procedimentos de eutanásia. Sessenta e cinco DVZs receberam o questionário online ou foram contatadas por telefone, dessas, trinta e três DVZs (50,8%) que responderam ao questionário foram incluídas no estudo e distribuídas em 11 mesorregiões para garantir anonimato. Os métodos de eutanásia reportados foram divididos em métodos químicos (injetáveis; inalatórios), físicos ou mistos. Os espécimes de morcegos (n = 550) foram identificados e classificados para avaliar os principais gêneros encontrados no estado. A localização da DVZ, as espécies, os hábitos alimentares e o método de eutanásia utilizado foram analisados. Os espécimes por família de espécies de morcegos foram Molossidae (n = 340), Phyllostomidae (n = 171) e Vespertilionidae (n = 39). Métodos químicos foram utilizados em 25 DVZs (75,75%), físicos em 5 (15,15%) e mistos em 3 (9,09%). Não há uniformidade ou padronização nos métodos de eutanásia de morcegos, embora a maioria seja baseada em métodos químicos aceitáveis. É proposto um algoritmo para auxiliar o veterinário na escolha do método de eutanásia, que permite a padronização desses procedimentos para os morcegos, considerando as diferenças fisiológicas e respeitando as diretrizes técnicas, bioéticas e de bem-estar animal.(AU)

Rabies virus isolated from insectivorous bats induces different inflammatory responses in experimental model

Rabies virus (RABV) is a neurotropic virus that causes fatal neuroinflammation in mammals. The insectivorous bat RABV strains are less pathogenic for mice than strains associated with other reservoirs. We characterized the tissue inflammatory response in the CNS of RABV isolated from insectivorous bats. Eptesicus furinalis (EPBRV)-infected mice had a robust inflammatory response and a greater amount of IL-1β, IL-6 and TNF-α, while Myotis nigricans (MNBRV)-infected mice showed a higher expression of IL-17 and greater activation of IFN-β. New approaches to understand the inflammatory response to different mechanisms of action may provide insights for the development of novel therapies for rabies.

Orthohepadnavirus infection in a neotropical bat (Platyrrhinus lineatus).

Hepatitis B virus (HBV) is the prototype of the Orthohepadnavirus genus and represents an important cause of chronic hepatitis, liver cirrhosis, and hepatic cancer in humans worldwide. To verify the occurrence and genetic variability of orthohepadnavirus among neotropical bats, we tested 81 liver samples of New World bats from São Paulo State, Southeastern Brazil, collected during 2012. PCR, sequencing, and phylogenetic analysis of Surface/Polymerase and Core viral genes confirmed the occurrence of the first isolate of bat orthohepadnavirus detected in South America. These results may contribute to subsequent studies of the origin, variability, host species, and evolution of bat orthohepadnaviruses in South America.

Hepatic lesions in captive owl monkeys (Aotus infulatus) with ultrasonographic "starry sky" liver.

BACKGROUND: The "starry sky" (SK) liver is ultrasonographic pattern characterized by multiple hyperechogenic foci in hepatic parenchyma. The study aimed to characterize the microscopic hepatic lesions in captive owl monkeys with SK liver. METHODS: Thirty-seven clinically healthy owl monkeys had their liver scanned and 18 of them had liver biopsy. Animals with SK and peliosis hepatis (PH) were subjected to immunohistochemical and molecular screening for Bartonella spp. RESULTS: SK liver occurred in 59.4% (22/37) of the owl monkeys. Biopsied animals showed steatosis, hydropic degeneration, hemosiderosis, PH, and multifocal granulomatous hepatitis. Two monkeys had SK, granulomatous hepatitis, and PH which were negative for Bartonella spp. CONCLUSIONS: PH and granulomatous hepatitis associated with hepatocellular degenerative lesions may present as hyperechoic nodular liver lesions consisted of SK liver; therefore, concomitant occurrence of two lesions or more contributed to the hepatic SK pattern among owl monkeys and such cases might be clinically monitored.

Evaluation of galectins 1 and 3 expression in canine melanoma / Avaliação da expressão das galectinas 1 e 3 no melanoma canino

Canine melanoma is a frequently-occuring neoplasm in dogs and presents as malignant and highly metastatic in this context, studies that contribute to the understanding of the tumor microenvironment in melanoma include the role of galectins. Galectins are proteins of the family of animal lectins that display carbohydrate recognition domains. Galectin-1 and galectin-3 are associated with neoplastic transformation, neoplastic cell survival, angiogenesis, immune system evasion, and metastasis. The goal of this study was to establish a correlation between expression patterns of galectin-1 and galectin-3 and the different degrees of aggressiveness of canine melanoma, as well as to determine serum concentration of galectin-3 in dogs with melanoma. Galectin-1 and galectin-3 expression was analyzed by immunohistochemistry in 30 canine melanomas, six melanocytomas and nine metastatic lymph nodes from patients whose primary tumors were also processed and analyzed. Serum samples from 30 dogs were collected and galectin-3 concentration was determined by ELISA and compared to the samples of 10 healthy dogs. Canine melanoma samples expressed galectin-1 in the cytoplasm and presented a variable pattern of galectin-3 staining depending on melanoma aggressiveness. We observed a decrease in the percentage of cells with cytoplasmic galectin-3 immunolabeling simultaneous to the increased nuclear staining intensity, while there was also a decrease in the percent frequency of nuclear galectin-3 immunolabeled cells according to progression of melanoma, comparing the least to the most aggressive cases. Dogs with melanoma had increased serum levels of galectin-3 when compared to healthy animals, suggesting its potential biomarker of patients with melanoma.(AU)

O melanoma canino é uma neoplasia frequente em cães que apresenta um potencial maligno e metastático. Neste contexto, investigar o microambiente tumoral é fundamental para compreender os mecanismos intercelulares e intracelulares envolvidos no desenvolvimento e progressão da doença. Neste estudo, destacamos as galectinas, proteínas da família das lectinas animais que exibem domínios de reconhecimento à carboidratos; a galectina-1 e a galectina-3 estão associadas a transformação neoplásica, sobrevivência de células neoplásicas, angiogênese, evasão do sistema immune e desenvolvimento de metástases. O objetivo deste estudo foi determinar os padrões de expressão de galectina-1 e galectina-3 em diferentes graus de agressividade do melanoma canino, bem como dosar a concentração sérica de galectina-3 em cães com melanoma e comparar com cães saudáveis. A expressão de galectina-1 e galectina-3 foi analisada em 30 melanomas caninos, seis melanocitomas e nove linfonodos metastáticos. A galectina-3 sérica foi mensurada em 30 cães com melanoma e comparada a 10 cães saudáveis. No melanoma canino a expressão de galectina-1 foi citoplasmática e a expressão de galectina-3 foi variável de acordo com o grau de agressividade. Notou-se uma redução na porcentagem de células com imunomarcação de galectina-3 citoplasmática e um aumento simultâneo da intensidade de imunomarcação nuclear, enquanto houve também uma diminuição na frequência percentual de células com imunomarcação nuclear de acordo com a progressão do melanoma comparando-se os casos menos com os mais agressivos. Cães com melanoma apresentaram níveis séricos aumentados de galectina-3 quando comparados a animais saudáveis, mostrando seu uso potencial como biomarcador em pacientes com melanoma.(AU)

Serological and molecular retrospective analysis of hepatitis E suspected cases from the Eastern Brazilian Amazon 1993-2014.

INTRODUCTION: We evaluated the anti-hepatitis E virus (HEV) antibody prevalence and HEV-RNA in archived serum samples of non-A-C hepatitis, or suspected cases of HEV infection from the Eastern Brazilian Amazon from 1993 to 2014. METHODS: Serum samples (n = 318) were tested using ELISA and immunoblotting, and screened for HEV-RNA by RT-qPCR. RESULTS: Anti-HEV IgM and IgG were detected in 3.4% (11/318) and 5.9% (19/318) of the samples, respectively. All samples were HEV-RNA negative. CONCLUSIONS: HEV was detected at a low prevalence. Broader serological and molecular evaluation of HEV infection in the Amazon region should be carried out.

A paruterinid metacestode in the liver of a Neotropical bat (Molossus molossus).

We describe a case of microscopic liver lesion caused by larval cestode in a frugivorous bat (Molossus molossus) from the Western Brazilian Amazon. Histopathological analysis of liver indicated the occurrence of metacestode associated with multifocal histiocytic response and the phylogenetic analysis of Cox-I and 18S rDNA genes indicated that the parasite belonged to the family Paruterinidae. This is the first identification of cestodes of the family Paruterinidae in bats and may suggest a broad range of paruterinid tapeworm hosts to be investigated in the Brazilian Amazon.

Using tilapia skin (Oreochromis niloticus) as an occlusive biological curative in equine wounds: short communication / Uso da pele de tilápia (Oreochromis niloticus) como curativo biológico oclusivo em feridas de equinos: nota prévia

Tilapia skin is being already use in humans and wild animals present burning wounds and showed a great result. The objective is to evaluate if tilapia skin used as an occlusive curative improves equine wound healing in two horses present chronic wound. Both animals are males, adults, both of breed Mangalarga Marchador South America. Every seven days wound we measured, photographed, biopsied for histopathological analysis, cleaned and tilapia curative was changed. Image J software was used to measure wound area. Tilapia skin as an occlusive biological factor seemed to improve healing process, wounds present an area reduction and clinical improvement during 35 days treatment, even though is still waiting for complete wound healing. In equine tilapia skin curative seemed to speed up healing process and allowed reduced curative change from every two days to once a week. This implies in decrease animal ́s stress, less pain and treatment cost reduction since we used less bandage amount. Beside that tilapia skin industrial waste. Furthermore, it allowed avoid using antibiotics, which reduces environment pollution and there ́s no antibiotic resistance issues.(AU)

A pele de tilápia está sendo utilizada em humanos e animais silvestres com feridas por queimadura demonstrando um excelente resultado. O objetivo do estudo é avaliar se a pele de tilápia utilizada como curativo oclusivo melhora o processo de cicatrização em dois equinos machos adultos da raça Mangalarga Machador, os quais apresentam feridas crônicas. A cada 7 dias as feridas eram medidas, fotografadas, biopsiadas para a análise histopatológica, limpas e o curativo de pele de tilápia trocado. O programa Image J foi utilizado para calcular a área da ferida. A pele de tilápia utilizada como curativo oclusivo parece ter um efeito positivo na cicatrização das feridas, a área diminuiu e a o aspecto clínico melhorou nos 35 dias avaliados, no entanto é necessário esperar pela cicatrização completa das feridas. Em equinos, a utilização de curativo de pele de tilápia parece diminuir o tempo de cicatrização e permite a redução do número de trocas de curativos de a cada dois dias para uma vez por semana. Isso implica em menos estresse e dor para o animal devido à menor manipulação e menor custo de tratamento, pois há menor quantidade de material de curativo sendo utilizado. Além disso, permite evitar a utilização de antibióticos, o que diminui o impacto ambiental e não gera resistência.(AU)

Serological and molecular retrospective analysis of hepatitis E suspected cases from the Eastern Brazilian Amazon 1993-2014

Abstract INTRODUCTION We evaluated the anti-hepatitis E virus (HEV) antibody prevalence and HEV-RNA in archived serum samples of non-A-C hepatitis, or suspected cases of HEV infection from the Eastern Brazilian Amazon from 1993 to 2014. METHODS Serum samples (n = 318) were tested using ELISA and immunoblotting, and screened for HEV-RNA by RT-qPCR. RESULTS Anti-HEV IgM and IgG were detected in 3.4% (11/318) and 5.9% (19/318) of the samples, respectively. All samples were HEV-RNA negative. CONCLUSIONS HEV was detected at a low prevalence. Broader serological and molecular evaluation of HEV infection in the Amazon region should be carried out.

Novel hybrid of Typical Enteropathogenic Escherichia coli and Shiga-Toxin-Producing E. coli (tEPEC/STEC) emerging from pet birds

Exotic psittacine birds have been implicated as reservoir of diarrheagenic Escherichia coli (E. coli), including enteropathogenic E. coli (EPEC) and Shiga-toxin producing E. coli (STEC). Here, we present a genotypic and phenotypic characterization of typical EPEC/STEC hybrid strains isolated from exotic psittacine birds. The strains were positive for eae, bfpA, and stx2f genes, belong to serotype O137:H6 and ST2678. Two strains were subject to whole genome sequencing, confirming the presence of the virulence factors of both E. coli pathotypes. Phenotypical in vitro tests confirmed their ability to adhere to HeLa cells and cause cytotoxicity to Vero cells. The rabbit ileal loop assays showed the attaching and effacing lesion, in addition to inflammatory process and overproduction of intestinal mucus. This is the first report of hybrid typical EPEC/STEC (O137:H6/ST2678) strains isolated from companion psittacine birds and the results suggest zoonotic risks.

Novel hybrid of Typical Enteropathogenic Escherichia coli and Shiga-Toxin-Producing E. coli (tEPEC/STEC) emerging from pet birds

Exotic psittacine birds have been implicated as reservoir of diarrheagenic Escherichia coli (E. coli), including enteropathogenic E. coli (EPEC) and Shiga-toxin producing E. coli (STEC). Here, we present a genotypic and phenotypic characterization of typical EPEC/STEC hybrid strains isolated from exotic psittacine birds. The strains were positive for eae, bfpA, and stx2f genes, belong to serotype O137:H6 and ST2678. Two strains were subject to whole genome sequencing, confirming the presence of the virulence factors of both E. coli pathotypes. Phenotypical in vitro tests confirmed their ability to adhere to HeLa cells and cause cytotoxicity to Vero cells. The rabbit ileal loop assays showed the attaching and effacing lesion, in addition to inflammatory process and overproduction of intestinal mucus. This is the first report of hybrid typical EPEC/STEC (O137:H6/ST2678) strains isolated from companion psittacine birds and the results suggest zoonotic risks.

The importance of clinical, histopathological and immunohistochemical marking for differential diagnosis of non-hematopoietic gastrointestinal mesenchymal neoplasms in dogs: literature review / Importância da avaliação clínica, anatomopatológica e imuno-histoquímica para o diagnóstico diferencial das neoplasias mesenquimais gastrointestinais não hematopoiéticas em cães: revisão de literatura

Gastrointestinal neoplasms in dogs represent a challenge for the veterinary clinician and surgeon as they are mostly malignant and when the owner notices the primary clinical changes the disease may already be in an advanced stage. Due to the high clinical and histopathological similarity between mesenchymal gastrointestinal neoplasms, it is often impossible to establish the definitive morphological diagnosis using light microscopy alone. In addition, there are only a few articles describing the anatomopathological and immunohistochemical characteristics, which make its complete characterization urgent and relevant in our context in order to assist the veterinary clinician, surgeon, and pathologist in establishing a precise diagnosis of these tumors.(AU)

As neoplasias gastrointestinais em cães representam casos desafiantes ao clínico e cirurgião, pois são em sua maioria malignas e, quando as primeiras alterações clínicas são notadas pelo proprietário do animal, a doença pode estar em estágio de desenvolvimento muito avançado. Devido à elevada similaridade clínica e histopatológica entre as neoplasias gastrointestinais mesenquimais, muitas vezes não é possível estabelecer o diagnóstico morfológico definitivo apenas com a microscopia de luz. Acrescenta-se que são poucos os artigos que descrevem as características anatomopatológicas e imuno-histoquímicas das neoplasias mesenquimais gastrointestinais que acometem os cães, o que torna premente a caracterização destas em nosso contexto, com o intuito de auxiliar clínico, cirurgião e patologista veterinário a estabelecer o diagnóstico destes tumores.(AU)

A survey on gram-negative bacteria in saffron finches (Sicalis flaveola) from illegal wildlife tradein Brazil / Pesquisa de bactérias gram-negativas em canários-da-terra (Sicalis flaveola) resgatados do tráfico ilegal de aves no Brasil

Passerines such as canaries or finches are the most unlawfully captured species that are sent to wildlife centers in São Paulo, Brazil. Captured birds may have infection by opportunistic bacteria in stressful situations. This fact becomes relevant when seized passerine are reintroduced. The aim of this study was to evaluate the health state of finches from illegal wildlife trade using microbiological approaches. Microbiological samples were collected by cloacal and tracheal swabs of 100 birds, captured during 2012 and 2013. The results indicate high frequency of gram-negative bacteria in feces and oropharynx, especially from the Enterobacteriaceae family (97.5%). The most frequent genera were Escherichia coli (46.5%) and Klebsiella pneumoniae (10.4%). Enterobacter cloacae, Serratia liquefaciens, Serratia spp. Klebsiella oxytoca and Citrobacter freundii were isolated with lower frequency from asymptomatic birds. The presence of enteropathogenic Escherichia coli (EPEC) and Shiga toxinproducing strain (STEC) confirm the zoonotic risks and public health concern.(AU)

No Estado de São Paulo, Brasil, os pássaros como os canários-da-terra têm sido uma das espécies mais frequentemente resgatadas do tráfico ilegal e enviadas aos centros de vida selvagem. Em situações de estresse estas aves podem ser acometidas por infecções causadas por bactérias oportunistas. Este fato é de grande importância quando é planejada da reintrodução das aves na natureza. O presente trabalho foi delineado para avaliar o estado de saúde de canários-da-terra resgatados do tráfico ilegal. Foram colhidas soabes da traqueia e da cloaca de 100 aves resgatadas durante os anos de 2012 e 2013. Os resultados obtidos revelaram alta frequência de bactérias gram-negativas nas fezes e no orofaringe dos animais, com maior frequência para os membros da família Enterobacteriaceae (97,5%). Os gêneros mais frequentes foram Escherichia coli (46,55) e Klebsiella pneumoniae (10,4%). Outros microorganismos incluindo Enterobacter cloacae, Serratia liquefaciens, Serratia spp, Klebsiella oxytoca e Citrobacter freundii também foram isolados em menor frequencia de aves assintomáticas. A presença de estirpes de Escherichia coli enteropagênicas (EPEC) e as produtoras da toxina de Shiga confirmam o risco de zoonose e a importância para saúde pública deste tipo de ave.(AU)