Resumo

This study aimed to determine the occurrence of anti-Toxoplasma gondii, Neospora caninum, and Leptospira spp. antibodies in sheep and goats raised in villages of the Xukuru do Ororubá indigenous community, Pernambuco, Brazil. A total of 180 serum samples from sheep and 108 serum samples from goats of both sexes and different ages were analyzed. For antibody research, indirect immunofluorescence antibody test (IFAT) were used for the protozoa T. gondii and N. caninum, and microscopic agglutination test (MAT) for Leptospira spp., with a cutoff titer of 1:64, 1:50 and 1:100, respectively. The frequency of anti-T. gondii antibodies was 16.6% (30/180) for sheep and 11.1% (12/108) for goats. The frequency of anti-N. caninum antibodies was 10.55% (19/180) for sheep, and 20.37% (22/108) for goats, while for Leptospira spp., 2.2% (4/180) of sheep and 1.85% (2/108) of goats reacted positively. The results obtained in this study are unprecedented in indigenous communities in the country and serve as an alert for monitoring goats and sheep from the Xukuru do Ororubá indigenous village regarding the occurrence and productive impact of infections by T. gondii, N. caninum, and Leptospira spp., in addition to the occurrence of the zoonosis toxoplasmosis and leptospirosis in the indigenous community.(AU)

Objetivou-se determinar a ocorrência de anticorpos anti-Toxoplasma gondii, Neospora caninum e Leptospira spp., em ovinos e caprinos criados em aldeias da comunidade indígena Xukuru do Ororubá, Pernambuco, Brasil. Foram analisadas 180 amostras de soro de ovinos e 108 amostras de soro de caprinos de ambos os sexos e diferentes idades. Para a pesquisa de anticorpos foi utilizada a técnica de Reação de Imunofluorescência indireta (RIFI), para os protozoários T. gondii e N. caninum e Aglutinação Microscópica (MAT) para Leptospira spp., com ponto de corte de 1:64, 1:50 e 1:100, respectivamente. A frequência de anticorpos anti-T gondii foi de 16,6% (30/180) em ovinos e 11,1% (12/108) em caprinos. A frequência de anticorpos anti-N. caninum foi de 10,55% (19/180) para ovinos e 20,37% (22/108) para caprinos, enquanto para Leptospira spp., 2,2% (4/180) dos ovinos e 1,85% (2/108) dos caprinos reagiram positivamente. Os resultados obtidos neste estudo são inéditos em comunidades indígenas do país e alertam para o monitoramento de caprinos e ovinos da aldeia indígena Xukuru do Ororubá, quanto à ocorrência e impacto produtivo de infecções por T. gondii, N. caninum e Leptospira spp., além da ocorrência de zoonoses como a toxoplasmose e leptospirose na comunidade indígena.(AU)

Resumo

Abstract Resveratrol, a natural polyphenol found in tempeh, has not been investigated especially in vitro as a neuroprotective agent against 2-methoxyethanol (2-ME)-induced beta-amyloid cytotoxicity. Beta amyloid peptides (A) could initiate neurotoxic events and neuron-inflammatory response via microglial activation. However, it remains unknown whether the neurotoxic effect of beta-amyloid and/or associated with the potential of 2-ME to induce neurotoxic effects on primary culture of nerve cells induced by 2-ME. This study investigated potential neuroprotective of trans-resveratrol a promising agent tempeh and soybean seed coats-derived against beta-amyloid cytotoxicity on primary culture of nerve cells induced by 2-methoxyethanol. Biotium and MTT assays were used to analyze neurons, which were isolated from the cerebral cortex of fetal mice at gestation day 19 (GD-19). A standard solution of 2-methoxyethanol was dosed at 10 L. The cultured cells were randomly divided into the following groups: (1) 2-ME group + resveratrol standard, (2) 2-ME group + resveratrol isolated from tempeh, (3) 2-ME group + resveratrol isolated from soybean seed coats, and (4) the control group, without the addition of either 2-ME or resveratrol. Exposure of the primary cortical neuron cells to beta-amyloid monoclonal antibody pre-incubated for 24 h with 10 µL of 4.2 µg/mL resveratrol and 7.5 mmol/l 2-methoxy-ethanol additions. Here, we report that the addition of 2-ME and resveratrol (standard and isolated from tempeh) of cell culture at concentrations of 1.4, 2.8 and 4.2 µg/mL showed that the majority of neurons grew well. In contrast, after exposure to 2-ME and Beta-amyloid, showed that glial activated. These findings demonstrate a role for resveratrol in neuroprotective-neurorescuing action.

Resumo O resveratrol, um polifenol natural encontrado em tempê, não foi investigado apenas in vitro como agente neuroprotetor contra a citotoxicidade beta-amiloide induzida por 2-metoxietanol (2-ME). Os peptídeos beta-amiloides (A) podem iniciar eventos neurotóxicos e resposta inflamatória dos neurônios via ativação microglial. No entanto, permanece desconhecido se o efeito neurotóxico do peptídeo beta-amiloide associado ao potencial do 2-ME causa efeitos neurotóxicos na cultura primária de células nervosas induzidas pelo 2-ME. Este estudo investigou o potencial neuroprotetor do agente trans-resveratrol em cascas de sementes de soja e tempê derivadas da citotoxicidade beta-amiloide na cultura primária de células nervosas induzidas pelo 2-metoxietanol. Ensaios de biotium e MTT foram utilizados para analisar os neurônios isolados do córtex cerebral de camundongos fetais no dia da gestação 19 (GD-19). As células cultivadas foram divididas aleatoriamente nos seguintes grupos: (1) grupo 2-ME + padrão de resveratrol; (2) grupo 2-ME + resveratrol isolado de tempê; (3) grupo 2-ME + resveratrol isolado de cascas de sementes de soja; e (4) grupo controle, sem a adição de 2-ME ou resveratrol. Houve exposição das células primárias dos neurônios corticais ao anticorpo monoclonal beta-amiloide pré-incubado por 24 horas, com 10 µL de 4,2 µg/mL de resveratrol, e adições de 7,5 mmol/l de 2-metoxietanol. A adição de 2-ME e resveratrol (padrão e isolado do tempê) da cultura de células nas concentrações de 1,4, 2,8 e 4,2 µg/mL mostrou que a maioria dos neurônios cresceu bem. Por outro lado, após a exposição ao 2-ME e beta-amiloide, a glia foi ativada. Esses achados demonstram um papel do resveratrol na ação neuroprotetora e de neurorresgate.

Resumo

Resveratrol, a natural polyphenol found in tempeh, has not been investigated especially in vitro as a neuroprotective agent against 2-methoxyethanol (2-ME)-induced beta-amyloid cytotoxicity. Beta amyloid peptides (Aβ) could initiate neurotoxic events and neuron-inflammatory response via microglial activation. However, it remains unknown whether the neurotoxic effect of beta-amyloid and/or associated with the potential of 2-ME to induce neurotoxic effects on primary culture of nerve cells induced by 2-ME. This study investigated potential neuroprotective of trans-resveratrol a promising agent tempeh and soybean seed coats-derived against beta amyloid cytotoxicity on primary culture of nerve cells induced by 2-methoxyethanol. Biotium and MTT assays were used to analyze neurons, which were isolated from the cerebral cortex of fetal mice at gestation day 19 (GD-19). A standard solution of 2-methoxyethanol was dosed at 10 μL. The cultured cells were randomly divided into the following groups: (1) 2-ME group + resveratrol standard, (2) 2-ME group + resveratrol isolated from tempeh, (3) 2-ME group + resveratrol isolated from soybean seed coats, and (4) the control group, without the addition of either 2-ME or resveratrol. Exposure of the primary cortical neuron cells to beta-amyloid monoclonal antibody pre-incubated for 24 h with 10 µL of 4.2 µg/mL resveratrol and 7.5 mmol/l 2-methoxy-ethanol additions. Here, we report that the addition of 2-ME and resveratrol (standard and isolated from tempeh) of cell culture at concentrations of 1.4, 2.8 and 4.2 µg/mL showed that the majority of neurons grew well. In contrast, after exposure to 2-ME and Beta-amyloid, showed that glial activated. These findings demonstrate a role for resveratrol in neuroprotective-neurorescuing action.

O resveratrol, um polifenol natural encontrado em tempê, não foi investigado apenas in vitro como agente neuroprotetor contra a citotoxicidade beta-amiloide induzida por 2-metoxietanol (2-ME). Os peptídeos beta amiloides (Aβ) podem iniciar eventos neurotóxicos e resposta inflamatória dos neurônios via ativação microglial. No entanto, permanece desconhecido se o efeito neurotóxico do peptídeo beta-amiloide associado ao potencial do 2-ME causa efeitos neurotóxicos na cultura primária de células nervosas induzidas pelo 2-ME. Este estudo investigou o potencial neuroprotetor do agente trans-resveratrol em cascas de sementes de soja e tempê derivadas da citotoxicidade beta-amiloide na cultura primária de células nervosas induzidas pelo 2-metoxietanol. Ensaios de biotium e MTT foram utilizados para analisar os neurônios isolados do córtex cerebral de camundongos fetais no dia da gestação 19 (GD-19). As células cultivadas foram divididas aleatoriamente nos seguintes grupos: (1) grupo 2-ME + padrão de resveratrol; (2) grupo 2-ME + resveratrol isolado de tempê; (3) grupo 2-ME + resveratrol isolado de cascas de sementes de soja; e (4) grupo controle, sem a adição de 2-ME ou resveratrol. Houve exposição das células primárias dos neurônios corticais ao anticorpo monoclonal beta-amiloide pré-incubado por 24 horas, com 10 µL de 4,2 µg/mL de resveratrol, e adições de 7,5 mmol/l de 2-metoxietanol. A adição de 2-ME e resveratrol (padrão e isolado do tempê) da cultura de células nas concentrações de 1,4, 2,8 e 4,2 µg/mL mostrou que a maioria dos neurônios cresceu bem. Por outro lado, após a exposição ao 2-ME e beta-amiloide, a glia foi ativada. Esses achados demonstram um papel do resveratrol na ação neuroprotetora e de neurorresgate.

Resumo

Resveratrol, a natural polyphenol found in tempeh, has not been investigated especially in vitro as a neuroprotective agent against 2-methoxyethanol (2-ME)-induced beta-amyloid cytotoxicity. Beta amyloid peptides (Aß) could initiate neurotoxic events and neuron-inflammatory response via microglial activation. However, it remains unknown whether the neurotoxic effect of beta-amyloid and/or associated with the potential of 2-ME to induce neurotoxic effects on primary culture of nerve cells induced by 2-ME. This study investigated potential neuroprotective of trans-resveratrol a promising agent tempeh and soybean seed coats-derived against betaamyloid cytotoxicity on primary culture of nerve cells induced by 2-methoxyethanol. Biotium and MTT assays were used to analyze neurons, which were isolated from the cerebral cortex of fetal mice at gestation day 19 (GD-19). A standard solution of 2-methoxyethanol was dosed at 10 µL. The cultured cells were randomly divided into the following groups: (1) 2-ME group + resveratrol standard, (2) 2-ME group + resveratrol isolated from tempeh, (3) 2-ME group + resveratrol isolated from soybean seed coats, and (4) the control group, without the addition of either 2-ME or resveratrol. Exposure of the primary cortical neuron cells to beta-amyloid monoclonal antibody pre-incubated for 24 h with 10 µL of 4.2 µg/mL resveratrol and 7.5 mmol/l 2-methoxy-ethanol additions. Here, we report that the addition of 2-ME and resveratrol (standard and isolated from tempeh) of cell culture at concentrations of 1.4, 2.8 and 4.2 µg/mL showed that the majority of neurons grew well. In contrast, after exposure to 2-ME and Beta-amyloid, showed that glial activated. These findings demonstrate a role for resveratrol in neuroprotective-neurorescuing action.

O resveratrol, um polifenol natural encontrado em tempê, não foi investigado apenas in vitro como agente neuroprotetor contra a citotoxicidade beta-amiloide induzida por 2-metoxietanol (2-ME). Os peptídeos betaamiloides (Aß) podem iniciar eventos neurotóxicos e resposta inflamatória dos neurônios via ativação microglial. No entanto, permanece desconhecido se o efeito neurotóxico do peptídeo beta-amiloide associado ao potencial do 2-ME causa efeitos neurotóxicos na cultura primária de células nervosas induzidas pelo 2-ME. Este estudo investigou o potencial neuroprotetor do agente trans-resveratrol em cascas de sementes de soja e tempê derivadas da citotoxicidade beta-amiloide na cultura primária de células nervosas induzidas pelo 2-metoxietanol. Ensaios de biotium e MTT foram utilizados para analisar os neurônios isolados do córtex cerebral de camundongos fetais no dia da gestação 19 (GD-19). As células cultivadas foram divididas aleatoriamente nos seguintes grupos: (1) grupo 2-ME + padrão de resveratrol; (2) grupo 2-ME + resveratrol isolado de tempê; (3) grupo 2-ME + resveratrol isolado de cascas de sementes de soja; e (4) grupo controle, sem a adição de 2-ME ou resveratrol. Houve exposição das células primárias dos neurônios corticais ao anticorpo monoclonal beta-amiloide pré-incubado por 24 horas, com 10 µL de 4,2 µg/mL de resveratrol, e adições de 7,5 mmol/l de 2-metoxietanol. A adição de 2-ME e resveratrol (padrão e isolado do tempê) da cultura de células nas concentrações de 1,4, 2,8 e 4,2 µg/mL mostrou que a maioria dos neurônios cresceu bem. Por outro lado, após a exposição ao 2-ME e beta-amiloide, a glia foi ativada. Esses achados demonstram um papel do resveratrol na ação neuroprotetora e de neurorresgate.

Resumo

ABSTRACT Purpose Quantify the tissue content of metalloproteinase-9 (MMP-9) and collagen in colic mucosa with and without intestinal transit after infliximab administration in rats subjected to Hartmann's surgery. Methods Twenty-two rats underwent colon diversion by Hartmann's surgery. Animals were maintained with intestinal bypass for 12 weeks to induce development of diversion colitis (DC). Afterwards, animals were divided into three groups: first group received subcutaneous application of saline solution (SS) 0.9%, while the remaining two groups received infliximab subcutaneously at doses of 5 or 10 mg·kg-1·week-1 for five consecutive weeks. After the intervention, animals were sacrificed, removing the segments with and without intestinal transit. Diversion colitis was diagnosed by histological study, and its intensity was determined by a validated inflammatory scale. Tissue expression of MMP-9 was assessed byimmunohistochemistry, while total collagen was assessed by histochemistry. Tissue content of both was measuredby computerized morphometry. Results Colon segments without intestinal transit had a higher degree of inflammation, which improved in animals treated with infliximab. Collagen content was always lower in those without intestinal transit. There was an increase in the collagen content in the colon without transit in animals treated with infliximab, primarily at a dose of 10 mg·kg-1·week-1. There was an increase in the content of MMP-9 in the colon without fecal transit, and a reduction was observed in animals treated with infliximab, regardless of the dose used. Conclusions Application of infliximab reduces inflammation, increases the total collagen content and decreases the content of MMP-9 in the colon without intestinal transit.

Resumo

Sperm sexing aims to separate sperm populations in carriers of the "X" or "Y" chromosome. Currently, flow cytometry is a technique that allows greater accuracy; however, it causes structural changes in sperm, reduces viability, and has a high cost. As a result, other methods have been researched, including immunosexing, which uses monoclonal antibodies to detect sex-specific surface antigens. Thus, the objective of this study was to evaluate the immunosexing technique using a monoclonal antibody against sex-specific protein (HY) in the conservation of ram and goat semen in ACP101/102c. Ejaculates from five rams and five goats were collected with the aid of an artificial vagina; they were evaluated and submitted to the immunosexing protocol, according to the manufacturer's recommendations, using the Monoclonal Antibody Kit specific for mammalian sperm with "Y" chromosomes (HY; HY Biotechnology, Rio de Janeiro, RJ, Brazil). After sexing, the supernatant was resuspended in the cryopreservation diluent: ACP ram (ACP101/102c + 20% egg yolk + 7% glycerol) and ACP goat (ACP101/102c + 2.5% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated at 4°C, stabilized for 30 min, frozen in liquid nitrogen vapor (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were evaluated in natura (T1), after immunosexing (T2) and after thawing (T3) for sperm motility subjectively using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique using eosin-nigrosine dye, and the percentages of healthy and morphologically defect spermatozoa were determined. In the evaluation of ram semen regarding sperm motility and IMP, no statistically significant differences were observed between treatments after sexing in the evaluation of absolute data (P > 0.05), with the difference being observed only between T1 and T2, and T3 (P < 0.05). Regarding the relative percentage and sperm morphology, no statistically significant differences were observed (P > 0.05). Regarding the evaluation of goat semen samples, the motility parameters were consistent with the technique submitted; however, the IMP data did not appear as expected, requiring further evaluation for a better assessment of the technique for this species. The data obtained from ram semen submitted to the immunosexing protocol, regarding the absolute evaluation of motility and IMP, demonstrated that the non-sexed semen (T1) was superior to the sexed treatments (T2 and T3); however, it is noteworthy that freezing started with approximately 50% of the cells, since the immunosexing technique results in a loss of viability of approximately 50% of the sperm, which corresponds to the ratio of sperm carrying the X chromosome. In addition, when the data in this study were transformed into relative values, no statistical differences were observed, indicating that the immunosexing protocol, as well as the freezing protocol, did not significantly affect the quality of ram sperm cells. In relation to the immunosexing of goat semen, future studies should be conducted in vitro to define a more appropriate protocol for the species and, in addition, in vivo studies should be performed to prove the quality of the technique. It was concluded that the immunosexing process using a monoclonal antibody against sex-specific protein (HY) associated with the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation of ovine species.(AU)

Resumo

Zika virus (ZIKV), an emerging arthropod-borne virus (arbovirus) of the Flaviviridae family, is a current issue worldwide, particularly because of the congenital and neurological syndromes associated with infection by this virus. As the initial clinical symptoms of all diseases caused by this group are very similar, clinical diagnosis is difficult. Furthermore, laboratory diagnostic efforts have failed to identify specific and accurate tests for each virus of the Flaviviridae family due to the cross-reactivity of these viruses in serum samples. This situation has resulted in underreporting of the diseases caused by flaviviruses. However, many companies developed commercial diagnostic tests after the recent ZIKV outbreak. Moreover, health regulatory agencies have approved different commercial tests to extend the monitoring of ZIKV infections. Considering that a specific and sensitive diagnostic method for estimating risk and evaluating ZIKV propagation is still needed, this review aims to provide an update of the main commercially approved serological diagnostics test by the US Food and Drug Administration (FDA) and Brazilian National Health Surveillance Agency (ANVISA). Additionally, we present the technologies used for monoclonal antibody production as a tool for the development of diagnostic tests and applications of these antibodies in detecting ZIKV infections worldwide.(AU)

Resumo

The identification of diversity of bovine pestiviruses circulating in the field is fundamental for continuous evaluation of diagnostic tests and vaccine composition. In this article we performed the genetic and antigenic characterization of twelve bovine pestiviruses isolated in the western region of Rio Grande do Sul, Brazil. The viruses were isolated from sera of bovine fetuses or from animals with clinical presentations suggestive of pestivirus infection. Genetic characterization by sequencing and phylogenetic analysis of the 5'UTR region of the viral genome allowed for the identification of bovine viral diarrhea virus (BVDV-1a, 4/12, 33.3%), BVDV-1b (6/12, 50%) and BVDV-2 (2/12, 16.7%). The reactivity of the isolates with a panel of monoclonal antibodies raised against envelope proteins (Erns, E1 and E2) demonstrated a high antigenic variability among isolates. Thus, the active circulation of bovine pestivirus infection, with high genetic and antigenic variability, in cattle on the western border of RS was confirmed, demonstrating the importance of continuous characterization of the pestiviruses circulating in the cattle herds to keep the diagnostic and control measures up to date.(AU)

A identificação da diversidade de pestivírus bovinos que circulam no campo é fundamental para a avaliação contínua dos testes de diagnóstico e composição de vacina. Neste artigo, realizamos a caracterização genética e antigênica de doze pestivírus bovinos isolados na região oeste do Rio Grande do Sul, Brasil. Os vírus foram isolados de soros de fetos bovinos ou de animais com apresentações clínicas sugestivas de infecção por pestivírus. A caracterização genética por sequenciamento e análise filogenética da região 5'UTR do genoma viral permitiu a identificação do vírus da diarréia viral bovina (BVDV-1a, 4/12, 33,3%), BVDV-1b (6/12, 50%) e BVDV-2 (2/12, 16,7%). A reatividade dos isolados com um painel de anticorpos monoclonais criados contra proteínas do envelope (Erns, E1 e E2) demonstrou uma alta variabilidade antigênica entre os isolados. Assim, confirmou-se a circulação ativa da infecção por pestivírus bovino, com alta variabilidade genética e antigênica, em bovinos na fronteira oeste do RS, demonstrando a importância da contínua caracterização dos pestivírus circulantes em bovinos para manter atualizadas as medidas de diagnóstico e controle.(AU)

Resumo

ABSTRACT: Feline leukemia virus (FeLV) causes an infection in cats that, in some cases, can also be reported with other pathologies, such as infection with feline immunodeficiency virus (FIV), feline infectious peritonitis (FIP), and lymphoma. Although, a compromised immune response is reported in these animals, little is known about the immunological state of their cells. To shed some light in this area, we studied peripheral blood samples from both infected and non-infected cats with FeLV, with or without FIV, FIP, and lymphoma. We tested a panel of monoclonal antibodies (n=11) against mouse and human antigens and we reported that cat leukocytes can be stained with anti-mouse B220 monoclonal antibody; therefore, percentages of B cells were evaluated in different cat groups. Our results showed that cats with FeLV and FIP, or with leukemia, presented a large decrease in B220+ mononuclear cells. However, FeLV+ cats without clinical signs, or with unspecific clinical signs, had the same amount of B220+ mononuclear cells as healthy cats (control cats). Since the expression of B220 is exclusively restricted to the naïve B cell population, we inferred that the absence of these B cells in FeLV+ cats is related to other conditions that affect B cell numbers, such as viral infections and leukemias. Therefore, the amount of naïve B cells in peripheral blood (i.e., B220+ cells) can be used to identify FeLV+ cats concomitantly carrying FIP or leukemia, from FeLV+ cats with lymphoma or without any clinical signs.

RESUMO: O vírus da leucemia felina (FeLV) causa de uma infecção em gatos, que também podem ter outras patologias, como a imunodeficiência felina (FIV), a peritonite infecciosa felina (FIP) e linfoma. Embora uma resposta imune comprometida seja encontrada nestes animais, pouco se sabe sobre o estado imunológico de suas células. Para ampliar o número de testes com a finalidade de avaliar o estado imunológico destes animais, estudamos amostras de sangue periférico de gatos infectados, ou não, com FeLV, e que apresentavam (concomitantemente) FIV, FIP e linfoma. Para isto, amostras de sangue foram marcadas com um painel de anticorpos monoclonais contra antígenos de camundongos e humanos (n = 11), para avaliar seu potencial para estudos imunológicos em gatos. De todo o painel de anticorpos testados, apenas o anticorpo anti-B220 de camundongo foi capaz de marcar leucócitos de gato. Nossos resultados mostraram que os gatos com FeLV e FIP, ou com leucemia, apresentaram uma grande diminuição nas células mononucleares B220+. No entanto, gatos FeLV+ sem sinais clínicos, ou com sinais clínicos inespecíficos, tiveram a mesma quantidade de células B220+ que os gatos saudáveis (gatos controle). Como a expressão de B220 é restrita à população de células B naïve, podemos inferir que a ausência dessas células B em gatos FeLV+ está relacionada a outras condições que afetam o número destas células, como infecções virais e leucemias. Portanto, a quantidade de células B naïve no sangue periférico pode ser usada para identificar gatos FeLV+ concomitantemente portadores de PIF ou leucemia, de gatos FeLV+ com linfoma ou sem sinais clínicos.

Resumo

The venom of bamboo vipers (Trimeresurus stejnegeri - TS), commonly found in Taiwan, contains deadly hemotoxins that cause severe envenomation. Equine-derived antivenom is a specific treatment against snakebites, but its production costs are high and there are some inevitable side effects. The aim of the present work is to help in the development of an affordable and more endurable therapeutic strategy for snakebites. Methods: T. stejnegeri venom proteins were inactivated by glutaraldehyde in order to immunize hens for polyclonal immunoglobulin (IgY) antibodies production. After IgY binding assays, two antibody libraries were constructed expressing single-chain variable fragment (scFv) antibodies joined by the short or long linker for use in phage display antibody technology. Four rounds of biopanning were carried out. The selected scFv antibodies were then further tested for their binding activities and neutralization assays to TS proteins. Results: Purified IgY from egg yolk showed the specific binding ability to TS proteins. The dimensions of these two libraries contain 2.4 × 107 and 6.8 × 107 antibody clones, respectively. An increase in the titers of eluted phage indicated anti-TS clones remarkably enriched after 2nd panning. The analysis based on the nucleotide sequences of selected scFv clones indicated that seven groups of short linkers and four groups of long linkers were identified. The recombinant scFvs showed significant reactivity to TS venom proteins and a cross-reaction to Trimeresurus mucrosquamatus venom proteins. In in vivo studies, the data demonstrated that anti-TS IgY provided 100% protective effects while combined scFvs augmented partial survival time of mice injected with a lethal amount of TS proteins. Conclusion: Chickens were excellent hosts for the production of neutralization antibodies at low cost. Phage display technology is available for generation of monoclonal antibodies against snake venom proteins. These antibodies could be applied in the development of diagnostic kits or as an alternative for snakebite envenomation treatment in the near future.(AU)

Resumo

Utilizando um anticorpo monoclonal contra a aflatoxina B1 (AFB1) como ligante, foi identificado um mimotopo específico de aflatoxina B1 após se realizarem quatro ciclos de seleção biológica de 7-peptídeos aleatórios em biblioteca de fago exibida. O mimotopo é denominado P10, e sua sequência de aminoácidos é YRRHEKD. O soro imunológico de ratos Balb/c imunizados com P10 foi especificamente ligado à aflatoxina B1-albumina, indicando que o anticorpo era específico ao AFB1. Esses resultados sugerem que é possível desenvolver a vacina baseada em mimotopo associado à toxina.(AU)

Resumo

As farmacodermias podem ser definidas como reações adversas em pele, mucosas e anexos, tendo, por vezes, caráter imunomediado. O diagnóstico baseia-se na avaliação clínico-laboratorial do paciente, envolvendo uma pesquisa acerca de fatores relacionados ao uso do fármaco e seus efeitos adversos. Na medicina veterinária, são escassos os relatos de reações farmacodérmicas. Logo, o objetivo do presente trabalho é relatar uma reação adversa após terapia otológica em cão. Foi atendido um paciente canino, fêmea, 02 anos de idade, com histórico de prurido auricular bilateral com evolução de três semanas. Na ocasião, foi realizado exame citológico auricular, evidenciando presença de elevada quantidade de células leveduriformes e cocos, além de células descamativas. Optou-se, então, por terapia à base de solução otológica composta por Gentamicina, Clotrimazol, Betametasona e Benzocaína. O quadro clínico evoluiu de forma satisfatória até o décimo dia de tratamento, quando a paciente apresentou intenso eritema e secreção melicérica bilateralmente. Repetiu-se o exame citológico, assim como realizou-se cultura de bactérias aeróbicas, sendo evidenciado em tais exames um infiltrado inflamatório piogranulomatoso, com pouca presença de conteúdo bacteriano e fúngico, corroborando com os achados da cultura bacteriana. Diante da suspeita de farmacodermia, procedeu-se com a troca de todos os compostos terapêuticos, tendo a paciente evoluído de forma satisfatória até o término do tratamento. Por tratar-se ainda de uma solução otológica composta, não se pode atribuir a causa da reação a especificamente um dos compostos. Contudo, reforça-se a necessidade de conscientização do médico veterinário acerca da identificação e adequada intervenção nas reações adversas medicamentosas, assim como espera-se sua contribuição científica na difusão dessas informações.

Pharmacodermia can be defined as adverse reactions in skin, mucous membranes and appendages, sometimes having immunomediated character. The diagnosis is based on the patient clinical-laboratorial evaluation, involving a research about factors related to the drug use and its adverse effects. In veterinary medicine, reports of pharmacodermic reactions are scarce. Therefore, the aim of the present study is to report an adverse reaction after otologic therapy in dogs. A 2-year-old female canine patient with a history of bilateral auricular pruritus with a three-week course was attended. At the time, auricular cytology was performed, evidencing the presence of high numbers of yeast cells and cocci, as well as desquamative cells. It was then opted for otologic solution composed of Gentamicin, Clotrimazole, Betamethasone and Benzocaine. The clinical presentation progressed satisfactorily until the tenth day of treatment, when the patient presented intense erythema and meliceric secretion. Cytological examination was repeated, as well as culture of aerobic bacteria. A piogranulomatous inflammatory infiltrate with low bacterial and fungal content was evidenced in these examinations, corroborating with the findings of the bacterial culture. Faced with the suspicion of pharmacodermia, all therapeutic compounds were exchanged, and the patient progressed satisfactorily until the end of the treatment. Because it is still a composed otological solution, the cause of the reaction cannot be attributed to specifically one of the compounds. However, there is a need to raise the awareness of the veterinarian about the identification and appropriate intervention in adverse drug reactions, as well as his scientific contribution to the dissemination of this information.

Resumo

Rabies is an important zoonosis with impact on livestock production. The diagnosis is usually based on the rabies virus detection in fresh or refrigerated brain samples by direct immunofluorescence (IF) performed on fixed brain smears. The aim of this study was to evaluate the performance of 6 monoclonal and 1 rabbit-polyclonal new antibodies for rabies diagnostic by using immunohistochemistry (IHC), which detects the agent on formalin fixed paraffin embedded tissues samples. Tested with 2 positive and 2 negative cows for rabies at dilutions 1/200 and 1/1000, obtained immunostaining was strong for one monoclonal and weak for the polyclonal at both dilutions. For 3 monoclonals, the immunostaining was weaker at 1/1000 and was negative at both dilutions for 2 monoclonals. Unwanted background was absent and negative samples remained clear for all antibodies. When the first monoclonal was applied on sections of brain of 19 cows with Rabies and 41 control cows at 1/1000 dilution, immunohistochemistry recognized all positive samples and was negative for all control cows. The number of cases analyzed did not allow estimating sensitivity and specificity of the tested assay, but the correlation observed between IF and IHC in both positive and negative samples suggested that accuracy of the test might be good. The results indicated that the tested antibody can detect the rabies virus on formalin fixed tissue samples, and that immunohistochemistry can complement other confirmatory tests when those cannot be performed.

Resumo

Abstract Purpose: To evaluate the effects of infliximab on the inflammation of the colonic mucosa devoid from fecal stream. Methods: Twenty-four rats were submitted to a Hartmann's procedure. They remained for 12 weeks with the fecal derivation to development of diversion colitis on excluded colorectal stump. After this period, they were divided into 3 groups: one group received intervention with saline (2.0 mL / week), other group infliximab at doses of 5 mg/kg/week and the other 10 mg/kg/week for five consecutively weeks. Concluded the intervention period, the animals were euthanized to remove colon segments with and without fecal stream. Colitis was diagnosed by histological analysis and the degree of inflammation by validated score. The neutrophilic infiltrate was evaluated by tissue expression of myeloperoxidase identified by immunohistochemical. The tissue content of myeloperoxidase was measured by computer-assisted image analysis. Results: The inflammatory score was high in colonic segments without fecal stream. The intervention with infliximab reduced the inflammatory score in excluded colonic segments. The content of myeloperoxidase was reduced in colonic segments of animals treated with infliximab mainly in high concentrations. Conclusion: Intervention with infliximab reduced the inflammation and the neutrophil infiltrate in colonic segments devoid of the fecal stream.

Resumo

This study describes a case of parvovirus infection in a river otter (Lontra longicaudis) assisted at the Wildlife Rehabilitation Center and Wildlife Screening Center, Federal University of Pelotas (UFPel), Rio Grande do Sul state, Brazil. Clinical signs included apathy, dark and fetid diarrhea, and crusted lesions on the palmar pads of the fore and hind limbs. The animal died after undergoing support treatment with antibiotics, anti-inflammatory, and fluid therapy. At necropsy, the intestines were reddened and edematous and the right kidney was diminished by one third of its normal size and covered with whitish, spongy material. A female Dioctophyma renale was found free in the abdominal cavity. Histologically, dilatation of the intestinal crypts and fusion and blunting of the intestinal villi were observed. In addition, moderate, multifocal lymphocytic enteritis with lymphoid depletion in Peyer's patches and mesenteric lymph nodes were present. Immunohistochemistry with anti-canine parvovirus monoclonal antibody (anti-CPV) was strongly positive in the bone marrow cells and enterocytes of the intestinal crypts, confirming the diagnosis of parvovirus infection. The peritoneum on the right kidney was expanded with a cuboidal cell border, forming multiple papillary projections associated with eggs of D. renale and severe inflammatory infiltrate (giant cells, macrophages, lymphocytes, eosinophils, and plasma cells). Areas of necrosis and mineralization were also observed. Due to fragmentation and degradation of its natural habitat, the otter approached the urban area and was contaminated with the virus, which is hosted and disseminated by domestic animals. Infection with D. renale can be associated with the large population of parasitized domestic animals, which eliminate the helminth eggs through urine, contaminating the environment where the parasite intermediate and paratenic hosts co-inhabit. The diseases of these animals can be a decline factor of wild populations that inhabit the region and are an alert to spillover risk.(AU)

Descreve-se um caso de parvovirose em uma lontra (Lontra longicaudis) enviada ao Núcleo de Reabilitação da Fauna Silvestre e Centro de Triagem de Animais Silvestres da Universidade Federal de Pelotas, Rio Grande do Sul, Brasil. O animal estava debilitado, apático, apresentava diarreia escura e fétida e lesões crostosas nos coxins palmares dos membros torácicos e pélvicos, morrendo após tratamento de suporte com antibiótico, anti-inflamatório e fluidoterapia. Na necropsia os intestinos estavam edematosos e avermelhados e o rim direito estava recoberto de material brancacento e esponjoso, com comprometimento de cerca de um terço do órgão. Foi observado, também, um exemplar de Dioctophyma renale, fêmea, livre na cavidade abdominal. Histologicamente havia fusionamento das vilosidades, dilatação das criptas intestinais com enterite linfocítica moderada multifocal e depleção linfoide nos linfonodos mesentéricos. Na técnica de imuno-histoquímica (IHQ) com anticorpo monoclonal anti-Parvovírus canino (Anti-CPV) houve marcação positiva nos enterócitos da base das vilosidades intestinais e na medula óssea, confirmando o diagnóstico de parvovirose. O peritônio sobre o rim direito estava espessado e revestido por células cuboides, formando múltiplas projeções papilares, nas quais observava-se acentuado infiltrado de células gigantes, macrófagos, linfócitos, eosinófilos e plasmócitos. Entre as projeções papilares havia ovos de Dioctophyma renale, áreas de necrose, calcificação e células gigantes. Conclui-se que a lontra, em função da fragmentação e degradação de seu habitat natural, aproximou-se do centro urbano e contaminou-se com o vírus, o qual é mantido e disseminado por animais domésticos. Por sua vez, a infecção por D. renale pode estar relacionada com a presença de animais domésticos parasitados, os quais eliminam ovos do helminto através da urina contaminando o ambiente, onde coabitam hospedeiros intermediários e paratênicos do parasito. As doenças desses animais podem ser um fator de declínio das populações de animais silvestres e alerta para o risco de spill-over na região.(AU)

Resumo

Despite common occurrence and importance of canine distemper disease the majority of tests currently available for diagnosis are hampered by either low sensitivity or specificity. In this study it was evaluated antigenic and immunogenic characteristics of a conserved region of nucleocapsid protein of canine distemper virus (rCDV NP) expressed in Escherichia coli employing a codon optimized synthetic gene. The expression of rCDVNP in Star strain (mean 300μg/mL, purified) was confirmed by SDS-PAGE and Western blot analysis by using His-Tag monoclonal antibodies. Western blot and ELISA, employing positive and negative control dog sera, demonstrated the rCDVNP antigenicity. The rCDVNP was inoculated in hens and immunoglobulin Y (IgY) was purified from the egg yolk. The mean yield of IgY was 28.55mg/mL. IgY reacted with the recombinant protein as demonstrated by Western blot and ELISA assays. In summary, our findings demonstrated that rCDVNP is antigenic since CDV positive dog sera recognized the protein in vitro. Additionally, the rCDVNP proved to be immunogenic in hens being possible to isolate a high concentration of specific IgY antibodies from the egg yolk. Taken together, these results indicate that the rCDVNP along with the specific IgY could be useful tools for development of the canine distemper immunodiagnostic assays.(AU)

Apesar da ocorrência comum e importância da cinomose canina, a maioria dos testes atualmente disponíveis para diagnóstico são prejudicados pela baixa sensibilidade ou especificidade. Neste estudo foram avaliadas características antigênicas e imunogênicas de uma região conservada da proteína do nucleocapsídeo do virus da cinomose canina (rCDV NP) expressa em Escherichia coli empregando um gene sintético e codons otimizados. A expressão na cepa Star (média de 300μg/mL, purificada) foi confirmada por SDS-PAGE e Western blot utilizando anticorpos monoclonais anti-His-Tag. A antigenicidade da rCDVNP foi demonstrada por western blot e ELISA empregando soros de cães positivos e negativos. A rCDVNP foi inoculada em galinhas e imunoglobulina Y (gY) foi obtida e purificada a partir da gema. A produção média de IgY foi 28.55mg/mL. Anticorpos IgY reagiram com a proteína recombinante, quando analisados por Western blot e ELISA. Em resumo, nossos achados demonstram que a rCDVNP produzida é antigênica, uma vez que os anticorpos de soro de cães positivos para CDV reconheceram a proteína in vitro. Além disso, a rCDVNP foi imunogênica em galinhas, sendo possível isolar anticorpos IgY específicos a partir da gema do ovo em altas concentrações. Tomados em conjunto, estes resultados indicam que a rCDVNP juntamente com a IgY específica podem ser ferramentas úteis para elaborar ensaios de imunodiagnóstico de cinomose canina.(AU)

Resumo

A case of blackleg in a brown brocket deer (Mazama gouazoubira) associated with trauma from being hit by a car in southern Rio Grande do Sul is reported. The clinical signs included fever, dehydration and lethargy that worsened progressively until 36 hours after the accident, when the animal died. In the fore right limb, there was a comminuted closed fracture of the radius and ulna but no skin wounds were observed. Grossly, the musculature of the pelvic limbs presented hemorrhage, edema and emphysema. Microscopically, the muscles of both rear legs had necrosis, edema, hemorrhage and mild inflammatory infiltration of neutrophils. Clostridium chauvoei was cultured from affected skeletal muscles, and it was also detected by immunohistochemistry, confirming a diagnosis of blackleg. The overlapping habitat of cattle and brown brocket deer is proposed as a predisposing factor in this case and alerts to spillover cases maybe happening in this region. In addition, blackleg should be included as differential diagnoses of deer with post-traumatic myositis.(AU)

Descreve-se um caso de carbúnculo sintomático em um veado-virá (Mazama gouazoubira), macho, jovem, resgatado após atropelamento em uma rodovia na região sul do Rio Grande do Sul. O cervídeo apresentou febre, desidratação e letargia, progredindo para a morte em 36 horas. No membro torácico direito foi observado fratura cominutiva fechada de rádio e ulna sem a presença de feridas perfurantes. Na necropsia foi observada hemorragia, edema e enfisema na musculatura dos membros pélvicos. Microscopicamente, os músculos dos membros pélvicos apresentaram necrose, edema, hemorragia e discreto infiltrado inflamatório neutrofílico. Houve o isolamento de Clostridium chauvoei e marcação positiva na técnica de IHQ com anticorpo monoclonal anti-C. chauvoei, confirmando o diagnóstico de carbúnculo sintomático. A sobreposição de habitat entre bovinos domésticos e cervídeos pode ser um fator de risco para esta doença e chama a atenção para casos de "spillover" que podem estar ocorrendo na região. Adicionalmente, sugere-se que o carbúnculo sintomático seja incluído nos diagnósticos diferenciais de cervídeos que apresentam miosite pós-traumática.(AU)

Resumo

The specificity of monoclonal antibodies (mAbs) to desired targets makes these molecules suitable for therapeutic and diagnostic uses against a wide range of pathogens. Phage display antibody libraries offer one method by which mAbs can be selected for, without the use of conventional hybridoma technology. In this work, phage display technology was used to construct, select and characterize a combinatorial single chain fragment variable (scFv) antibody library against bovine herpesvirus type 1 (BoHV-1) from the immune repertoire of chickens immunized with the virus. In silico analysis of the hypervariable domains of the antibody heavy chains revealed a high frequency of scFv fragments with low variability, suggesting that selection had probably been carried out and favored by a few im-munogenic viral antigens. The reactivity of the scFv fragments selected against BoHV-1 was demon-strated by Phage-ELISA. A significant increase in antibody reactivity to the target was observed after six rounds of library selection, showing its potential use as a molecule for BoHV-1 diagnosis. The strategy described here opens up a field for the use of phage display as a tool for selection of mono-clonal antibodies that could be used for theranostic applications against infectious and parasitic dis-eases of veterinary interest.

A especificidade dos anticorpos monoclonais (mAb) aos alvos desejados torna estas moléculas ade-quadas para uso em diagnóstico ou terapia de uma vasta gama de agentes patogênicos. Biblioteca de anticorpos apresentados em fagos filamentosos é uma metodologia para a produção de mAbs, poden-do ser utilizada como alternativa à tecnologia de hibridoma convencional, tradicionalmente empregada para este fim. Neste trabalho, a tecnologia de Phage display foi usada para construir, selecionar e ca-racterizar uma biblioteca combinatorial de fragmentos de anticorpos de cadeia única (scFv) contra o Herpesvírus bovino tipo 1 (BoHV-1) a partir do repertório imune de galinhas imunizadas com o vírus. A análise in silico dos domínios hipervariáveis das cadeias pesadas dos anticorpos revelou uma alta frequência de fragmentos scFv com baixa variabilidade, sugerindo que a seleção foi provavelmente conduzida e favorecida por poucos antígenos virais mais imunogênicos. A reatividade dos fragmentos scFv selecionados contra BoHV-1 foi demonstrada por Fago-ELISA. Observou-se um aumento signi-ficativo da reatividade dos anticorpos após seis ciclos de seleção, evidenciando sua utilização como molécula para o diagnóstico de BoHV-1. A estratégia aqui descrita abre a possibilidade do uso da tec-nologia de Phage display como ferramenta na seleção de anticorpos monoclonais com potencial uso tanto para o diagnóstico quanto para terapia de doenças infecciosas e/ou parasitárias de interesse veterinário.

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Background: Paracoccidioidomycosis (PCM) is a neglected systemic mycosis caused by a dimorphic fungus of the Paracoccidioides genus. The standard diagnosis is based on isolation of the fungi in culture, and by microscopic visualization of characteristic multiple budding yeast cells in biological samples. However, in some situations, access to the site of injury prevents the collection of biological material. A variety of immuno-serological techniques has proven useful for allowing inferring diagnosis with a certain degree of certainty, thus optimizing time. The aim of this study was to standardize and validate the Dot-ELISA (DE) assay, comparing it with the serological standard, double immunodiffusion (DI). Methods: In order to standardize the DE assay, 143 serum samples were used. Out of those, 23 were from apparently healthy patients, 77 were from patients with confirmed PCM and 43 were from patients with other lung infections (tuberculosis, aspergillosis and histoplasmosis). To validate the DE technique, 300 serum samples from patients with PCM clinical suspicion (probable and possible cases) were employed, and these results were compared with those of DI. Results: The DE assay showed sensitivity of 91%, specificity of 95.4%, positive predictive value of 96%, negative predictive value of 98.2%, accuracy of 93%, and great precision (k = 0.93). In addition, the nitrocellulose membranes have proved to be viable for using at least 90 days after P. brasiliensis B-339 antigen sensitization. Conclusion: Dot-ELISA method was found to be an extremely promising tool as serologic screening technique, because of its high sensitivity. Furthermore, Dot-ELISA shows the prospect of being transferred to laboratories of mycoserology including those with fewer resources or even to be used directly in the field. It has an excellent shelf life ­ membranes coated with antigen can be used for testing without changes in the pattern of reactivity among laboratories ­ and presents reliable values of sensitivity, specificity, predictive values, accuracy and a high correlation with the serological standard methodology. Based on the present findings, it possible to state that this technique constitutes a remarkable option to be used in routine diagnosis for public health centers.(AU)

Resumo

Esta tese é constituída por três capítulos compostos por um artigo científico cada, produzidos através de pesquisas sobre doenças e lesões do sistema cardiovascular de cães na região do semiárido Nordestino. O primeiro artigo intitulado Aspectos patológicos das doenças cardiovasculares em cães, diagnosticadas no Semiárido Paraibano. Objetivou-se descrever as doenças e lesões que afetam o sistema cardiovascular de em cães. Foram incluídos no presente estudo os cães que apresentaram diagnóstico necroscópico de afecção cardíaca. Adicionalmente, os casos em que não havia diagnóstico de doença cardíaca foram reavaliadas fotografias e lâminas histopatológicas. Adicionalmente, foram extraídos dos arquivos os dados referentes aos protocolos histoquímicos e imuno-histoquímicos utilizados para a confirmação de agentes infecciosos. Para a caracterização morfológica dos fungos, foi utilizada coloração de metenamina nitrato de prata de Grocott (GMS) e para confirmação do fungo envolvido foi realizada imuno-histoquímica utilizando anticorpo monoclonal específico para Rhizopus arrhizus (Clone WSSA-RA-1, AbD Serotec, Bio-Rad®, São Paulo, Brasil); anticorpo policlonal anti-Coccidioides immitis (Gibson Bioscience®, Lexyngton, KY) e o anticorpo policlonal anti- Candida albicans (Abcam ab53891®). Para os casos de amebas de vida livre, foi utilizado anticorpo primário policlonal anti-Balamuthia, Acanthamoeba e Naegleria com soro de coelho. Nos casos de parvovirose cardíaca, foi utilizado como anticorpo primário, utilizou-se anticorpo monoclonal anti-parvovirus. Para os actinomicetos foram realizadas as colorações de Ziehl- Neelsen modificado, Brown-Brenn modificado e Giemsa. Adicionalmente, foram obtidos dados dos cultivos bacteriológicos. Após essa classificação, foi calculada a prevalência de cada grupo e de cada condição em relação ao total de animais necropsiados. De 2513 cães necropsiados, 210 animais apresentaram alterações cardiovasculares. Essas afecções foram caracterizadas em degenerativas adquiridas, infecciosas (bacterinas, virais e fúngicas), parasitárias (protozoóticas e helmínticas), metabólicas (mineralização), neoplásicas (primárias e metastáticas), traumáticas e malformações. O segundo artigo, intitulado Aspectos epidemiológicos e patológicos das infecções fúngicas cardíacas em cães no Semiárido Nordestino. Objetivou-se com este estudo descrever os fatores de risco relacionado ao desenvolvimento dessas afecções, juntamente com os aspectos anatomopatológicos das lesões observadas nos cães afetados. Foram realizadas técnicas de histoquímica e imuno-histoquímica para a descrição dos aspectos morfotintoriais e identificação dos agentes fúngicos, respectivamente. No período de estudo, que correspondeu de 2003 a 2019, cinco cães apresentaram infecção fúngica sistêmica com envolvimento cardíaco. Os agentes fúngicos diagnosticados foram Candida albicans; Coccidioides immitis e Rhizopus arrhizus. No ponto de vista epidemiológico, notou-se que no período da ocorrência dessas infecções ocorrem em uma época do ano muito similar, sugerindo fortemente que além das condições de imunossupressão que levaram a infecção, a sazonalidade pode colaborar para uma maior disponibilidade de formas infectantes de agentes fúngicos no ambiente. O terceiro artigo, Aspectos epidemiológicos, clínicos e patológicos da endocardite vegetativa em cães. Objetivou-se com este estudo descrever os fatores associados ao desenvolvimento da endocardite em cães, bem como os aspectos fisiopatológicos. No período de 2003 a 2019, foram diagnosticados 19 casos de endocardite em cães. De acordo com o tipo de endocardite, foram categorizadas em endocardite bacteriana, urêmica e marântica. Os eventos tromboembólicos relacionados a endocardite bacteriana demonstraram-se ser mais frequentes do que os outros tipos de endocardite.

This thesis consists of three chapters composed of one scientific article each, produced through research on diseases and injuries of the cardiovascular system of dogs in the semi-arid region of the Northeast. The first article entitled Pathological aspects of cardiovascular diseases in dogs, diagnosed in the semiarid region of Paraiba (2003-2019). The objective was to describe the diseases and injuries that affect the cardiovascular system in dogs. Dogs with a necroscopic diagnosis of cardiac disease were included in the present study. Additionally, in cases where there was no diagnosis of heart disease, photographs and histopathological slides were reassessed. Additionally, data referring to the histochemical and immunohistochemical protocols used for the confirmation of infectious agents were extracted from the files. For the morphological characterization of the fungi, Grocott's silver nitrate methenamine stain (GMS) was used and to confirm the fungus involved, immunohistochemistry was performed using a monoclonal antibody specific for Rhizopus arrhizus (Clone WSSA-RA-1, AbD Serotec, Bio - Rad®, São Paulo, Brazil); anti-Coccidioides immitis polyclonal antibody (Gibson Bioscience®, Lexyngton, KY) and anti-Candida albicans polyclonal antibody (Abcam ab53891®) (Alves et al., 2019). For free-living amoeba cases, primary antibody was used. polyclonal anti-Balamuthia, Acanthamoeba and Naegleria with rabbit serum (Frade et al., 2015). In cases of cardiac parvovirus, anti-parvovirus monoclonal antibody was used as the primary antibody. The technique was described by Souto et al., (2018). For Actinomycetes, modified Ziehl-Neelsen, modified Brown-Brenn and Giemsa stains were performed. Additionally, data from bacteriological cultures were obtained. After this classification, the prevalence of each group and each condition was calculated in relation to the total number of necropsied animals. Of 2513 necropsied dogs, 210 animals showed cardiovascular changes. These conditions were characterized as degenerative acquired, infectious (bacterin, viral and fungal), parasitic (protozootic and helminthic), metabolic (mineralization), neoplastic (primary and metastatic), traumatic and malformations. The second article, entitled Epidemiological and pathological aspects of cardiac fungal infections in dogs in the Northeastern Semiarid Region. The objective of this study was to describe the risk factors related to the development of these conditions, along with the anatomopathological aspects of the lesions observed in affected dogs. Histochemical and immunohistochemical techniques were performed for the description of morphotintorial aspects and identification of fungal agents, respectively. In the study period, which corresponded from 2003 to 2019, five dogs had systemic fungal infection with cardiac involvement. The fungal agents diagnosed were Candida albicans; Coccidioides immitis and Rhizopus arrhizus. From an epidemiological point of view, it was noted that during the period of occurrence of these infections they occur at a very similar time of year, strongly suggesting that in addition to the immunosuppression conditions that led to the infection, seasonality may contribute to a greater availability of infective forms of infection. fungal agents in the environment. The third article, Clinical, epidemiological, pathological aspects of vegetative endocarditis in dogs. The objective of this study was to describe the factors associated with the development of endocarditis in dogs, as well as the pathophysiological aspects. In the period from 2003 to 2019, 19 cases of endocarditis were diagnosed in dogs. According to the type of endocarditis, they were categorized into bacterial, uremic and marantic endocarditis. Thromboembolic events related to bacterial endocarditis have been shown to be more frequent than other types of endocarditis.