Cytotoxic effects of crotoxin from Crotalus durissus terrificus snake in canine mammary tumor cell lines.

Background: Mammary gland tumors are the most prevalent neoplasm in intact female dogs, and they are good natural models to study comparative oncology. Most canine mammary malignancies, as in women, are commonly refractory to conventional therapies and demand continuous new therapeutic approaches. Crotalus durissus terrificus, also called rattlesnake, has more than 60 different proteins in its venom with multiple pharmaceutical uses, such as antitumor, antiviral, and antimicrobial action. Crotoxin, a potent ß-neurotoxin formed by the junction of two subunits, a basic subunit (CB-PLA2) and an acidic subunit (crotapotin), has already been reported to have anticancer properties in different types of cancers. Methods: In this work, we describe the cytotoxic potential of crotoxin and its subunits compared to doxorubicin (drug of choice) in two canine mammary carcinoma cell lines. Results: Crotoxin, CB-PLA2, crotalic venom, and doxorubicin decreased cell viability and the ability to migrate in a dose-dependent manner, and crotapotin did not present an antitumoral effect. For all compounds, the predominant cell death mechanism was apoptosis. In addition, crotoxin did not show toxicity in normal canine mammary gland cells. Conclusion: Therefore, this work showed that crotoxin and CB-PLA2 had cytotoxic activity, migration inhibition, and pro-apoptotic potential in canine mammary gland carcinoma cell lines, making their possible use in cancer research.

Cytotoxic effects of crotoxin from Crotalus durissus terrificus snake in canine mammary tumor cell lines

Background: Mammary gland tumors are the most prevalent neoplasm in intact female dogs, and they are good natural models to study comparative oncology. Most canine mammary malignancies, as in women, are commonly refractory to conventional therapies and demand continuous new therapeutic approaches. Crotalus durissus terrificus, also called rattlesnake, has more than 60 different proteins in its venom with multiple pharmaceutical uses, such as antitumor, antiviral, and antimicrobial action. Crotoxin, a potent β-neurotoxin formed by the junction of two subunits, a basic subunit (CB-PLA2) and an acidic subunit (crotapotin), has already been reported to have anticancer properties in different types of cancers. Methods: In this work, we describe the cytotoxic potential of crotoxin and its subunits compared to doxorubicin (drug of choice) in two canine mammary carcinoma cell lines. Results: Crotoxin, CB-PLA2, crotalic venom, and doxorubicin decreased cell viability and the ability to migrate in a dose-dependent manner, and crotapotin did not present an antitumoral effect. For all compounds, the predominant cell death mechanism was apoptosis. In addition, crotoxin did not show toxicity in normal canine mammary gland cells. Conclusion: Therefore, this work showed that crotoxin and CB-PLA2 had cytotoxic activity, migration inhibition, and pro-apoptotic potential in canine mammary gland carcinoma cell lines, making their possible use in cancer research.

Detection of Equus caballus Papillomavirus in Equine Aural Plaque Samples.

Aural plaques have been linked to Equus caballus papillomavirus (EcPV). Ten types of EcPVs have already been described; however, only EcPVs 1, 3, 4, 5, and 6 have been observed in association with aural plaques. Accordingly, the objective of this study was to evaluate the presence of EcPVs in equine aural plaque samples. A total of 29 aural plaque samples (from 15 horses) were collected and assessed for the presence of the DNA of these EcPVs by PCR. Additionally, 108 aural plaque samples used in previous research were evaluated for the presence of EcPVs 8 and 9. Previously described primers were used for PCR to detect EcPVs 1 to 8, and specific primers were designed for EcPV 9. Minigenes were synthesized and used as a positive control in the PCRs for the undetected EcPVs. EcPVs 2, 7, 8, and 9 were not detected in any of the evaluated samples, suggesting that these viral types are not involved in the etiology of the equine aural plaque in Brazil. EcPV 6 was the most prevalent (81%), followed by EcPVs 3 (72%), 4 (63%) and 5 (47%), which reinforces the idea that these viruses play an important role in the etiology of the equine aural plaque in Brazil.

Ligamentum teres reconstruction using autogenous semitendinosus tendon with toggle technique in rabbits.

Background: Ligamentum teres (LT) has traditionally been considered a vestigial or redundant structure in humans; however, based on new studies and the evolution of hip arthroscopy, the LT injury has been viewed as a source of hip pain. Therefore, LT reconstruction can be beneficial in some cases. Rabbits have been frequently used as a model for cranial cruciate ligament reconstruction but few studies are available for ligamentum teres reconstruction. Objective: To evaluate the semitendinosus tendon to replace ligamentum teres with the toggle technique, using rabbits as an experimental model. Methods: Twenty-six female Norfolk rabbits with approximately 3 months of age were divided into two equal groups after excision of ligamentum teres (LT) from the right hip joint: G1-no reconstruction of LT and capsulorrhaphy; G2-double-bundle reconstruction of the LT using semitendinosus tendon autograft. In both groups, the LT was removed from the right hip joint. In G2 the autograft was harvested from the left hind limb of the same rabbit. The rabbits were evaluated clinically at different time intervals; before surgery (M1), 48 h (M2), 15 days (M3), 30 days (M4) and 90 days (M5) after surgery. Results: The rabbits supported their limbs on the ground in both the groups. As complications of the procedure, four hip joints showed subluxations in the radiographic evaluation of G1; three at M4 and one at M5. In G2; two luxations of hip joints at M3 and one subluxation at M4 were seen. On ultrasound, irregular articular surface was seen in 30.8% of the rabbits that had subluxation of hip joints. Gross evaluation identified tendon graft integrity in 76.92% of the rabbits. Histological analysis revealed graft adhesion to the bone in the early phase comprised of sharpey-like collagen fibers. Conclusion: The double-bundle reconstruction of the LT using autologous semitendinosus tendon associated with the toggle rod shows an early phase of tendon graft ligamentization at 90 days post-operatively in young rabbits, but biomechanical bias suffered by the tendon during gait must be considered.

Immunohematological features of free-living Alouatta belzebul (Linnaeus, 1766) red-handed howler monkeys in the Eastern Amazon.

The red-handed howler monkey (Alouatta belzebul) is one of the 35 threatened Brazilian primate species found in two highly endangered Brazilian biomes. Their Amazonian native populations have been declining due to exponential deforestation associated with human activities, especially the construction of dams. The studied population (n = 27) was located in the Belo Monte dam Area of Influence. For the first time, we presented hematological parameters and the basic profile of T (CD3) and B (BSAP PAX5) cells by immunocytochemistry. The results supported the hypothesis that the immuno-hematological profile is influenced by sex, age, and season. Eosinophils were significantly higher in females (p = 0.03), monocytes statistically greater in juveniles (p = 0.04), and total plasma protein increased significantly (p > 0.001) during the dry season. Furthermore, adults showed a statistically higher average absolute number of B lymphocytes than young individuals (p = 0.03), in contrast to T lymphocytes. Even without knowing the full history of antigenic exposure, these results not only contribute to elucidating the boundaries between health and disease but may help lay the groundwork for future research into the effects of anthropogenic stress on immune activation.

Epidemiological characterization of reported cases of brucellosis in cattle in the western region of the state of Santa Catarina, Brazil / Caracterização epidemiológica dos casos notificados de brucelose em bovinos na região Oeste do Estado de Santa Catarina, Brasil

ABSTRACT: Brucellosis is one of the most important and widespread bacterial zoonoses in the world. There are few epidemiological studies on brucellosis in cattle in the productive region of western Santa Catarina. Thus, the objective of this study was to characterize the epidemiological aspects of bovine brucellosis reported in this region from 2013 to 2018. Cattle data were provided by the Integrated Company for Agricultural Development of Santa Catarina (Companhia Integrada de Desenvolvimento Agrícola de Santa Catarina - CIDASC). The prevalence of the disease was evaluated in cattle and herds by means of temporal trend analysis and spatial distribution. Bovines with tests showing positive for brucellosis were analyzed for location (municipality) and year of confirmation of diagnosis. In the study period (2013-2018), 686,086 tests were performed for brucellosis, with a prevalence rate of 1.11% (7,586/686,086). Among the 80,171 herds tested, 0.95% were found to be positive (764/80171). The odds ratio of brucellosis occurrence in cattle increased significantly in 2015 (OR = 1.55) and 2016 (OR = 1.97) and decreased in 2014 (OR = 0.47) and 2017 (OR = 0.97). The disease is present in cattle in 61.07% (80/131) of the municipalities where the tests were performed. Results showed that brucellosis is endemic in the region.

RESUMO: A brucelose é uma das zoonoses bacterianas mais importantes e difundidas no mundo. São poucos os estudos epidemiológicos sobre a doença em bovinos na região produtiva do Oeste de Santa Catarina. Desta forma, o objetivo deste estudo foi caracterizar os aspectos epidemiológicos da brucelose bovina notificados nesta região entre os anos de 2013 a 2018. Os dados sobre os bovinos foram fornecidos pela Companhia Integrada de Desenvolvimento Agrícola de Santa Catarina (CIDASC). Foram avaliadas as prevalências da doença nos bovinos e rebanhos, por meio de análise de tendência temporal e distribuição espacial. Os bovinos, cujo exame confirmatório mostrou positividade para brucelose, foram analisados quanto à localização (município) e o ano de confirmação do diagnóstico. No período de estudo (2013-2018) foram realizados 686.086 testes para brucelose, encontrando-se prevalência de 1,11% (7.586/686.086) de animais positivos, enquanto a prevalência de rebanhos positivos, dentre os 80.171 testados, foi de 0,95% (764/80171). As razões de chances de ocorrência de brucelose nos bovinos aumentaram significativamente nos anos de 2015 (OR= 1,55) e 2016 (OR= 1,97) e diminuíram em 2014 (OR= 0,47) e 2017 (OR= 0,97). A doença está presente nos bovinos em 61,07% (80/131) dos municípios em que foram realizados os testes. Os resultados demonstram que a brucelose é endêmica na região.

Molecular Expression Profile Reveals Potential Biomarkers and Therapeutic Targets in Canine Endometrial Lesions.

Cystic endometrial hyperplasia (CEH), mucometra, and pyometra are common uterine diseases in intact dogs, with pyometra being a life threatening disease. This study aimed to determine the gene expression profile of these lesions and potential biomarkers for closed-cervix pyometra, the most severe condition. Total RNA was extracted from 69 fresh endometrium samples collected from 21 healthy female dogs during diestrus, 16 CEH, 15 mucometra and 17 pyometra (eight open and nine closed-cervixes). Global gene expression was detected using the Affymetrix Canine Gene 1.0 ST Array. Unsupervised analysis revealed two clusters, one mainly composed of diestrus and CEH samples and the other by 12/15 mucometra and all pyometra samples. When comparing pyometra with other groups, 189 differentially expressed genes were detected. SLPI, PTGS2/COX2, MMP1, S100A8, S100A9 and IL8 were among the top up-regulated genes detected in pyometra, further confirmed by external expression data. Notably, a particular molecular profile in pyometra from animals previously treated with exogenous progesterone compounds was observed in comparison with pyometra from untreated dogs as well as with other groups irrespective of exogenous hormone treatment status. In addition to S100A8 and S100A9 genes, overexpression of the inflammatory cytokines IL1B, TNF and IL6 as well as LTF were detected in the pyometra from treated animals. Interestingly, closed pyometra was more frequently detected in treated dogs (64% versus 33%), with IL1B, TNF, LBP and CXCL10 among the most relevant overexpressed genes. This molecular signature associated with potential biomarkers and therapeutic targets, such as CXCL10 and COX2, should guide future clinical studies. Based on the gene expression profile we suggested that pyometra from progesterone treated dogs is a distinct molecular entity.

Nodular dermatofibrosis in a German shepherd dog: case report.

An 7-year-old male German shepherd dog not neutered was attended presenting several cutaneous nodules ranging from 0.5-3 cm in diameter at least a one year history, located mainly in thoracic and pelvic limbs, with progressive weight loss over two months. Fine needle aspiration biopsy and pelvic and thoracic limbs nodules excision biopsy were performed. After histopathological diagnosis, nodular dermatofibrosis diagnosis was determined and abdominal ultrasound was performed for possible renal cysts evaluation. The animal received treatment to control secondary bacterial infection. It is necessary to carry out histopathological examination and ultraosund to identify possible renal cysts and for definitive diagnosis. There is no specific treatment for nodular dermatofibrosis.

Estudo da leishmaniose visceral canina no município de Goiânia, Goiás, Brasil / Studies on canine visceral leishmaniasis in the municipality of Goiânia, Goiás, Brazil

A leishmaniose visceral (LV) é uma zoonose causada por Leishmania infantum chagasi. Canídeos desempenham importante papel epidemiológico por atuarem como reservatórios naturais. Este trabalho teve como objetivo conduzir uma investigação sobre leishmaniose visceral canina (LVC) em cães na cidade de Goiânia-GO. Para o estudo, foram utilizados 214 animais sintomáticos e assintomáticos e colhidas amostras de sangue, biópsias de pele e aspirados de linfonodo. Nos testes sorológicos pela reação de imunofluorescência indireta (RIFI), 20 animais apresentaram resultados positivos. No exame parasitológico direto, foram detectadas amastigotas em seis amostras. Os testes pela reação em cadeia da polimerase (PCR), realizados apenas nos animais sintomáticos ou com resultado positivo em pelo menos um dos exames anteriores (n igual 81), foram positivos para sete cães. As amostras positivas no exame parasitológico foram também submetidas à PCR. Das 20 amostraspositivas nos testes pela RIFI, apenas sete foram confirmadas pela PCR. Os resultados positivos pela PCR espécie-específica, nas sete amostras, confirmaram a identidade molecular de L. i. chagasi nestes animais. Os resultados registraram os primeiros casos autóctones em Caldas Novas-GO e Novo Brasil-GO e evidenciaram a necessidade urgente de intensificação das estratégias de controle da LVC nestes municípios. O estudo revelou a existência de casos de LVC em cães que buscam atendimento clínico na cidade de Goiânia.