Testes bioquímicos para identificação de leveduras na medicina veterinária - revisão de literatura / Biochemical tests to yeast identification in veterinary medicine - literature review / Pruebas bioquímicas para identificación de levaduras en la medicina veterinaria - revisión de literatura

As leveduras são fungos de importância à medicina veterinária por causarem doenças infecciosas em diferentes hospedeiros animais. A presente revisão de literatura teve como objetivo relatar os principais testes bioquímicos capazes de auxiliar na identificação de fungos leveduriformes de interesse veterinário e zoonótico. Para o levantamento bibliográfico, foram consideradas 48 publicações científicas selecionadas na área e indexadas nas principais bases de dados, entre os anos de 1988 e 2020. Como resultados, observou-se que oito provas são as mais empregadas na rotina micológica. Devido à baixa variabilidade morfológica das espécies leveduriformes, testes bioquímicos complementares são fundamentais na rotina laboratorial. A análise do perfil bioquímico de leveduras contribui na determinação taxonômica dos fungos a partir de reações químicas, visto que o metabolismo varia de acordo com a espécie, resultando em metabólitos distintos, os quais podem ser avaliados por diferentes provas. Conclui-se que a identificação fenotípica das leveduras é imprescindível no diagnóstico, prognóstico, tratamento e controle de doenças fúngicas e contribui para a manutenção da saúde animal.(AU)

Yeasts are fungi of importance to veterinary medicine because they cause infectious diseases in different animal hosts. This literature review aimed to report the main biochemical tests capable of assisting in the identification of yeast-like fungi of veterinary and zoonotic interest. For the bibliographical survey, 48 selected scientific publications in the area and indexed in the main databases, between the years 1988 and 2020, were considered. As a result, it was observed that eight tests are the most used in the mycological routine. Due to the low morphological variability of yeast species, complementary biochemical tests are fundamental in the laboratory routine. The analysis of the biochemical profile of yeast contributes to the taxonomic determination of fungi based on chemical reactions, since the metabolism varies according to the species, resulting in different metabolites, which can be evaluated by different tests. It is concluded that the phenotypic identification of yeasts is essential in the diagnosis, prognosis, treatment and control of fungal diseases and contributes to the maintenance of animal health.(AU)

Las levaduras son hongos de importancia para la medicina veterinaria porque causan enfermedades infecciosas en diferentes animales huéspedes. Esta revisión de la literatura tuvo como objetivo informar las principales pruebas bioquímicas capaces de ayudar en la identificación de hongos tipo levadura de interés veterinario y zoonótico. Para el levantamiento bibliográfico se consideraron 48 publicaciones científicas seleccionadas en el área e indexadas en las principales bases de datos, entre los años 1988 y 2020. Como resultado se observó que ocho pruebas son las más utilizadas en la rutina micológica. Debido a la baja variabilidad morfológica de las especies de levaduras, las pruebas bioquímicas complementarias son fundamentales en la rutina del laboratorio. El análisis del perfil bioquímico de la levadura contribuye a la determinación taxonómica de los hongos en base a reacciones químicas, ya que el metabolismo varía según la especie, dando como resultado diferentes metabolitos, los cuales pueden ser evaluados mediante diferentes pruebas. Se concluye que la identificación fenotípica de levaduras es fundamental en el diagnóstico, pronóstico, tratamiento y control de enfermedades fúngicas y contribuye al mantenimiento de la salud animal.(AU)

Biochemical parameters, C-reactive protein, and proteinogram of Sapajus libidinosus kept in captivity / Parâmetros bioquímicos, proteína C-reativa e proteinograma de Sapajus libidinosus mantido em cativeiro

Sapajus libidinosus is a New World primate belonging to the Cebidae family that lives in the caatinga and cerrado, which are known Brazilian biomes. It is currently classified as near threatened, almost endangered, by the main animal protection organizations. Knowledge of biochemistry, the C-reactive protein, and the proteinogram are important for the preservation of this species. Our study established reference intervals for several biochemical variables and the proteinogram. For this purpose, blood samples were collected from 50 S. libidinosus monkeys (25 males and 25 females) kept in captivity at the Brazilian state of Paraíba. Descriptive analysis and reference intervals were estimated following the guidelines of the American Society of Veterinary Clinical Pathology, using the Reference Value Advisor 2.1 software. In the overall population (n=50), significant differences (P < 0.05) were noted for creatinine and total proteins when considering the age factor (higher in adults), as well as for albumin and indirect bilirubin (higher in juveniles). Sex-associated differences (females versus males, P < 0.05) were reported for serum urea and creatinine (higher in males), as well as for albumin and Beta-Globulins (higher in females). In conclusion, to the authors' best knowledge, the present results are innovative and can be used as reference intervals for assessing the animals' health status. Moreover, it is also pioneer in determining the C-reactive protein in this species. It is important to emphasize that gender and age categories can have an influence and should be considered when interpreting the tests results.

O Sapajus libidinosus é um primata do novo mundo pertencente à família Cebidae que habita os biomas da caatinga e cerrado do Brasil. Atualmente esta classificado como quase ameaçado pelos principais orgãos de proteção animal. O conhecimento da bioquímica sérica, proteína C reativa e proteinograma são importantes na preservação desta espécie. O nosso estudo tem como objetivo determinar os intervalos de referência para várias variáveis bioquímicas e proteinograma. Para isso, foram colhidas amostras de sangue de 50 macacos da espécie S. libidinosus (25 machos e 25 fêmeas) mantidos em cativeiro no estado da Paraíba, Brasil. A análise descritiva e os cálculos dos intervalos de referência foram estimados segundo as diretrizes da Sociedade Americana de Patologia Clínica Veterinária com o uso do software Reference Value Advisor 2.1. Na população total (n=50), considerando o factor idade, foram observadas diferenças significativas (P < 0,05) para a creatinina e proteínas totais (valores superiores nos adultos) e para a albumina e bilirrubina indirecta (valores superiores nos jovens). Foram encontradas diferenças associadas ao sexo (fêmeas versus machos, P < 0,05) para a ureia e creatinina séricas (valores superiores nos machos) e albumina e Beta-Globulinas (valores superiores nas fêmeas). Conclui-se que, tanto quanto é do conhecimento dos autores, os resultados deste estudo são inovadores e podem ser utilizados como intervalos de referência para avaliação do estado de higidez dos animais e é pioneiro na determinação da proteína C reativa nesta espécie. É importante salientar que o sexo e a faixa etária podem ter influência e devem ser levados em consideração pelos profissionais na interpretação dos exames.

CA 15-3, CRP, and LDH correlates with prognostic parameters in canine mammary neoplasms

The identification of putative prognostic factors in canine mammary neoplasms (CMNs) has been focused on tissue-specific biomarkers, but the serum biomarkers, including cancer antigen 15-3 (CA 15-3), c-reactive protein (CRP), and lactate dehydrogenase (LDH) have been demonstrated to display clinical application in cases of CMNs. The aim of the study was to evaluate the levels of these serum biomarkers and their association with well-established prognostic factors in CMNs. Samples from 15 female canines with CMNs and 15 clinically healthy ones were collected. The results were evaluated using the Tukey's, Pearson, or Spearman tests. The cut-off point, sensitivity, specificity, and area under curve (AUC) were evaluated using the receiver operating characteristic (ROC) curve analysis in a logistic regression model (P<0.05). The levels of CA 15-3, CRP and LDH were significantly higher in the serum of female dogs with CMNs compared to the healthy ones. Moreover, these factors were positively correlated with ulceration, tumor size, histopathological grade, metastatic lymph node, and clinical staging. Female dogs with CMNs were found to exhibit highest serum levels of CA 15-3, CRP, and LDH. Therefore, they can be applied to improve the efficacy of the diagnosis and prognostic evaluation in casas of CMNs.(AU)

Padronização de um teste ELISA indireto para diagnóstico da cinomose canina / Standardization of an indirect ELISA test for canine diagnosis diagnosis

A cinomose é uma enfermidade causada pelo vírus Canine Distemper Virus (CDV). Essa doença afeta principalmente cães, mas também acomete outras espécies domésticas e selvagens. A imunidade do animal está relacionada ao grau que a esse patógeno vai atingir o organismo do indivíduo. Ela afeta a respiração do animal, pode causar vômito, diarreia, convulsões, podendo levar o animal à óbito. O objetivo do presente trabalho foi padronizar um teste ELISA indireto com antígeno de superfície para o diagnostico cinomose utilizando amostras de soro canino. Para padronização da técnica, fez-se necessário o estudo da diluição do antígeno para identificar a melhor concentração para sensibilização da placa. O teste foi aplicado primeiramente com diferentes diluições do antígeno para detecção do melhor desempenho do antígeno. Feito isso, foi testado em um banco de soro de 45 animais comprovadamente positivos no teste ELISA comercial e em soro de 45 animais comprovadamente negativos no teste ELISA comercial, posteriormente foi calculado o ponto de corte, especificidade e sensibilidade do teste. O teste ELISA indireto se mostrou com excelência como um teste de diagnóstico para a cinomose canina, obtendo-se ponto de corte de densidade óptica de 0,229, sensibilidade de 95,5% e especificidade de 84,4%.

Distemper is a disease or the disease by the CDV virus, Distemper Virus. This disease mainly affects dogs, but also affects other domestic and wild species. The animal's immunity is related to the degree to which it will reach the individual's organism. It affects the animal's breathing, can cause vomiting, diarrhea, convulsions, and can lead to death. The aim of the present work test was to standardize an indirect ELISA for distemper diagnosis in experiments using a surface antigen. For the study of technical identification, it was necessary to specify the antigen for the best concentration of plaque sensitization. The test was initially applied with different dilutions of the antigen to detect the best performance of the antigen. This was tested in a serum bank of 45 animals proven positive in the commercial ELISA test and in the serum of 45 animals proven negative in the commercial ELISA test, later it was tested on the cut-off point, specificity and sensitivity of the test. The indirect ELISA test proved to be excellent as a diagnostic test for canine distemper, with an optical density cut-off of 0.229, sensitivity of 95.5% and specificity of 84.4% being obtained.

Multiparametric assay of screening for the diagnosis of mycoses of interest in Public Health: standardization of methodology / Ensaio multiparamétrico de triagem para diagnóstico de micoses de interesse em Saúde Pública: padronização da metodologia

The standardization and validation of a multiplex assay requires the combination of important parameters such as sensitivity and specificity, acceptable levels of performance, robustness, and reproducibility. We standardized a multiparametric Dot-blot aimed at the serological screening of paracoccidioidomycosis, histoplasmosis, and aspergillosis. A total of 148 serum were evaluated: 10 from healthy subjects, 36 from patients with paracoccidioidomycosis, 62 from patients with histoplasmosis, and 40 from patients with aspergillosis. It was found that the multiparametric Dot-blot showed a high percentage of cross-reactivity. However, when evaluated individually, in the serological screening of histoplasmosis, a good performance was observed when compared to the double immunodiffusion assay, considered the gold standard test, with 100% co-positivity and 83.3% co-negativity. The performance of serological screening for aspergillosis was not satisfactory when compared to double immunodiffusion, showing 71.4% co-positivity and 100% co-negativity. The evaluation of the stability of nitrocellulose membranes showed that membranes sensitized with H. capsulatum antigen remained stable for 90 days and those sensitized with A. fumigatus antigen for 30 days. We conclude that the use of crude antigens was not suitable for the standardization of the multiparametric Dot-blot assay, due to the high cross-reactivity, and that further tests should be performed with purified proteins (AU).

A padronização e validação de um ensaio multiplex requer a combinação de parâmetros importantes, como sensibilidade e especificidade, níveis aceitáveis de desempenho, robustez e reprodutibilidade. Este trabalho padronizou um Dot-blot multiparamétrico visando a triagem sorológica da paracoccidioidomicose, histoplasmose e aspergilose. Foram avaliadas 148 amostras de soro: 10 de indivíduos saudáveis, 36 de pacientes com paracoccidioidomicose, 62 de pacientes com histoplasmose e 40 de pacientes com aspergilose. Verificou-se que o Dot-blot multiparamétrico apresentou elevado percentual de reatividade cruzada. Entretanto, quando avaliado individualmente, na triagem sorológica da histoplasmose observou-se bom desempenho quando comparado ao ensaio de imunodifusão dupla, considerado o teste padrão ouro, com 100% de co-positividade e 83,3% de co-negatividade. O desempenho da triagem sorológica da aspergilose não foi satisfatório quando comparado a imunodifusão dupla, apresentando 71,4% de co-positividade e 100% de co-negatividade. A avaliação da estabilidade das membranas de nitrocelulose mostrou que membranas sensibilizadas com antígeno de H. capsulatum permaneceram estáveis por 90 dias e as sensibilizadas com antígeno de A. fumigatus, por 30 dias. Concluímos que o uso de antígenos brutos não foi adequado para a padronização do ensaio de Dot-blot multiparamétrico, devido ao alto índice de reatividade cruzada, e que novos testes devem ser realizados com proteínas purificadas (AU).

Acute phase protein and vitamin D concentration in dogs with multicentric lymphoma / Concentração de proteínas de fase aguda e vitamina D em cães com linfoma multicêntrico

This study aimed to evaluate the serum concentration of vitamin D (25-Hydroxyvitamin D) and acute phase proteins (APPs; alpha-1 acid glycoprotein, haptoglobin, transferrin, ceruloplasmin, albumin, IgA, IgG and alpha-1 - antitrypsin) as potential biomarkers for prognostic and therapy response in dogs with multicentric lymphoma submitted to the CHOP (Cyclophosphamide, Doxorubicin, Vincristine and Prednisone) chemotherapy protocol. Thirteen dogs with multicentric lymphoma classified as high grade by cytology were included in the treatment group (GL), while ten healthy dogs were included in the control group (GC). Serum was collected in the weeks T0, T5 and T10 of CHOP chemotherapy protocol, for the GL group, and in a single collection, for the GC group. All the collected samples were evaluated for the APPs and vitamin D concentrations through electrophoresis and chemiluminescence methods, respectively. Diagnostic and staging tests were performed for all the dogs in the GL group, and included cytopathology, histopathology and immunohistochemistry of the affected lymph node. Of these dogs, 9 achieved a complete response and 4 a partial response to the treatment. Data analysis was performed with the R software. The results demonstrated that serum concentrations of IgA, haptoglobin and α1-acid glycoprotein were significantly different between the groups and also between the different chemotherapy times analyzed (p<0.05), indicating that these proteins can be considered as sensitive biomarkers for lymphoma in dogs. Furthermore, the α1-acid glycoprotein showed prognostic value for the disease, with 63% specificity. However, vitamin D concentration was not correlated with prognosis of the dogs with lymphoma.

Objetivou-se caracterizar a concentração sérica da vitamina D e das PFAs (Proteínas de Fase Aguda) (alfa-1 glicoproteína ácida, haptoglobina, transferrina, ceruloplasmina, albumina, IgA, IgG e alfa-1 ­ antitripsina) em cães com linfoma multicêntrico, submetidos ao tratamento quimioterápico com protocolo CHOP (Ciclofosfamida, Doxorrubicina, Vincristina e Prednisona), determinando o valor prognóstico desses marcadores para a doença. Foram avaliadas as concentrações séricas das PFAs, através do método da eletroforese e as concentrações da vitamina D, através da quimioluminescência em dois grupos experimentais, um grupo de 13 cães com linfoma multicêntrico classificados como alto grau pela citologia (GL) durante as semanas T0, T5 e T10 do tratamento com protocolo quimioterápico antineoplásico e em um grupo de 10 animais saudáveis para compor o grupo controle (GC), em coleta única. Para isso, foi realizado o diagnóstico, estadiamento e avaliação de resposta terapêutica dos 13 pacientes com linfoma multicêntrico através de técnicas de citopatologia, histopatologia, imuno-histoquímica do linfonodo periférico acometido. Foi observado que 9 pacientes tiveram resposta completa e 4 pacientes tiveram resposta parcial ao tratamento. Os dados foram analisados através do software R. Os resultados indicam que as diferenças entre as variáveis IgA, haptoglobina e α1-glicoproteína ácida foram significativas entre os grupos, e entre os diferentes momentos da quimioterapia (p< 0,05), indicando que podem ser marcadores sensíveis ao linfoma em cães. A α1-glicoproteína ácida apresentou valor prognóstico para o linfoma, com 63% de especificidade. Porém a vitamina D não apresentou valor prognóstico para o linfoma multicêntrico em cães.

Pseudopterígio em coelho - tratamento com tracolimus / Pseudopterygium in a rabbit - treatment with tracolimus

Background: Pseudopterygium, also known as aberrant conjunctival growth, is poorly described in the literature, althoughit is known that this abnormality is uncommon and affects dwarf rabbits and their crossbreeds. The etiology of this diseaseis unknown, but there are hypotheses that the conjunctival growth cause may have its origins in immunological factors,inflammation, traumatic conditions, or cartilage dysplasias. Thus, this study reports the treatment efficacy applied in arabbit, through the continuous use of tracolimus eye drops, after surgical procedure of conjunctival fold resection, as away of controlling the pseudopterygium in rabbits.Case: This case report discusses the positive results from the surgical and therapeutic conduct of a clinical case attended bythe Ophthalmology and Microsurgical Veterinary Service at the Hospital Veterinário Universitário (HVU) of the UFSM. Thepatient was a male rabbit, sterilized, approximately 2-year-old, crossed with a dwarf rabbit. The owner’s main complaintwas the change in the aspect of the left eye, with progressive worsening in the previous four weeks. In the ophthalmologicalexamination, the animal did not present impaired vision or discomfort, however, a vascularized pink membrane was noted,which consisted of a fold of the bulbar conjunctiva, that grew centripetally and covered 90% of the cornea in 360 degrees.The diagnosis was confirmed through visual inspection and the patient’s history. The eye alteration had a characteristicaspect, described as proliferation of the bulbar conjunctiva over the cornea, in a centripetal manner and without signs ofinflammation. In addition, other ophthalmological alterations were ruled out during the patient’s physical and specificexamination. The patient was referred for anesthetic evaluation and, in addition, pre-surgical blood tests were performed,which were normal, according to the expected ranges for the species...

Pseudopterígio em coelho - tratamento com tracolimus / Pseudopterygium in a rabbit - treatment with tracolimus

Background: Pseudopterygium, also known as aberrant conjunctival growth, is poorly described in the literature, althoughit is known that this abnormality is uncommon and affects dwarf rabbits and their crossbreeds. The etiology of this diseaseis unknown, but there are hypotheses that the conjunctival growth cause may have its origins in immunological factors,inflammation, traumatic conditions, or cartilage dysplasias. Thus, this study reports the treatment efficacy applied in arabbit, through the continuous use of tracolimus eye drops, after surgical procedure of conjunctival fold resection, as away of controlling the pseudopterygium in rabbits.Case: This case report discusses the positive results from the surgical and therapeutic conduct of a clinical case attended bythe Ophthalmology and Microsurgical Veterinary Service at the Hospital Veterinário Universitário (HVU) of the UFSM. Thepatient was a male rabbit, sterilized, approximately 2-year-old, crossed with a dwarf rabbit. The owners main complaintwas the change in the aspect of the left eye, with progressive worsening in the previous four weeks. In the ophthalmologicalexamination, the animal did not present impaired vision or discomfort, however, a vascularized pink membrane was noted,which consisted of a fold of the bulbar conjunctiva, that grew centripetally and covered 90% of the cornea in 360 degrees.The diagnosis was confirmed through visual inspection and the patients history. The eye alteration had a characteristicaspect, described as proliferation of the bulbar conjunctiva over the cornea, in a centripetal manner and without signs ofinflammation. In addition, other ophthalmological alterations were ruled out during the patients physical and specificexamination. The patient was referred for anesthetic evaluation and, in addition, pre-surgical blood tests were performed,which were normal, according to the expected ranges for the species...(AU)

Cloning and expression of Neospora caninum rhoptry 2 and its potential as an immunobiological for neosporosis control / Clonagem e expressão de rhoptry 2 Neospora caninum e seu potencial como um imunobiológico para o controle de neosporose

The protozoan Neospora caninum is known worldwide as one of the main causes of abortion in cattle. During infection, rhoptry proteins present in the apical complex of the parasite play important roles in adhesion and parasitophorous vacuole formation. The use of N. caninum ROP2 in experimental vaccines has shown promising protective results. In our study we performed cloning and expression in Escherichia coli of an antigenic portion of N. caninum ROP2. The recombinant protein (rROP2) was obtained in insoluble form, and the purified protein showed a size of approximately 18kDa. Even being a small truncate NcROP2 region, it was possible to conserve the antigenic epitopes which were recognized by bovine serum naturally infected with N. caninum. Vaccination with rROP2 on aluminum hydroxide adjuvant induced high levels of rROP2-specific IgG antibodies capable of recognizing native protein in tachyzoite lysates. In conclusion, our approaches were effective in obtaining the rROP2 protein, which induced specific mouse immune response and was also recognized by sera from N. caninum naturally infected cattle. These results suggest that it is a promising antigen for the development of neosporosis subunit vaccines as well as a suitable antigen for use in immunodiagnosis.(AU)

O protozoário Neospora caninum é conhecido mundialmente como uma das principais causas de aborto em bovinos. Durante a infecção, as proteínas rhoptry presentes no complexo apical do parasita desempenham papel importante na adesão e formação de vacúolos parasitóforos. O uso de ROP2 de N. caninum em vacinas experimentais tem mostrado resultados de proteção promissores. Em nosso estudo, realizamos a clonagem e expressão em Escherichia coli de uma porção antigênica de N. caninum ROP2. A proteína recombinante (rROP2) foi obtida na forma insolúvel, e a proteína purificada apresentou tamanho aproximado de 18kDa. Mesmo sendo uma pequena região truncada de NcROP2, foi possível conservar os epítopos antigênicos que foram reconhecidos pelo soro de bovinos naturalmente infectados com N. caninum. A vacinação com rROP2 adsorvida no adjuvante de hidróxido de alumínio induziu altos níveis de anticorpos IgG anti-rROP2, capazes de reconhecer a proteína nativa em lisados de taquizoítos. Em conclusão, nossas abordagens foram eficazes na obtenção da proteína rROP2, que induziu resposta imune específica em camundongos e também foi reconhecida por soros de bovinos naturalmente infectados com N. caninum. Estes resultados sugerem que rROP2 é um antígeno promissor para o desenvolvimento de vacinas de subunidades de neosporose, bem como um antígeno adequado para uso em imunodiagnóstico.(AU)

Cloning and expression of Neospora caninum rhoptry 2 and its potential as an immunobiological for neosporosis control / Clonagem e expressão de rhoptry 2 Neospora caninum e seu potencial como um imunobiológico para o controle de neosporose

The protozoan Neospora caninum is known worldwide as one of the main causes of abortion in cattle. During infection, rhoptry proteins present in the apical complex of the parasite play important roles in adhesion and parasitophorous vacuole formation. The use of N. caninum ROP2 in experimental vaccines has shown promising protective results. In our study we performed cloning and expression in Escherichia coli of an antigenic portion of N. caninum ROP2. The recombinant protein (rROP2) was obtained in insoluble form, and the purified protein showed a size of approximately 18kDa. Even being a small truncate NcROP2 region, it was possible to conserve the antigenic epitopes which were recognized by bovine serum naturally infected with N. caninum. Vaccination with rROP2 on aluminum hydroxide adjuvant induced high levels of rROP2-specific IgG antibodies capable of recognizing native protein in tachyzoite lysates. In conclusion, our approaches were effective in obtaining the rROP2 protein, which induced specific mouse immune response and was also recognized by sera from N. caninum naturally infected cattle. These results suggest that it is a promising antigen for the development of neosporosis subunit vaccines as well as a suitable antigen for use in immunodiagnosis.(AU)

O protozoário Neospora caninum é conhecido mundialmente como uma das principais causas de aborto em bovinos. Durante a infecção, as proteínas rhoptry presentes no complexo apical do parasita desempenham papel importante na adesão e formação de vacúolos parasitóforos. O uso de ROP2 de N. caninum em vacinas experimentais tem mostrado resultados de proteção promissores. Em nosso estudo, realizamos a clonagem e expressão em Escherichia coli de uma porção antigênica de N. caninum ROP2. A proteína recombinante (rROP2) foi obtida na forma insolúvel, e a proteína purificada apresentou tamanho aproximado de 18kDa. Mesmo sendo uma pequena região truncada de NcROP2, foi possível conservar os epítopos antigênicos que foram reconhecidos pelo soro de bovinos naturalmente infectados com N. caninum. A vacinação com rROP2 adsorvida no adjuvante de hidróxido de alumínio induziu altos níveis de anticorpos IgG anti-rROP2, capazes de reconhecer a proteína nativa em lisados de taquizoítos. Em conclusão, nossas abordagens foram eficazes na obtenção da proteína rROP2, que induziu resposta imune específica em camundongos e também foi reconhecida por soros de bovinos naturalmente infectados com N. caninum. Estes resultados sugerem que rROP2 é um antígeno promissor para o desenvolvimento de vacinas de subunidades de neosporose, bem como um antígeno adequado para uso em imunodiagnóstico.(AU)

In vitro and in vivo genotoxicity and cytotoxicity analysis of protein extract from Aplysina fulva sponges

This study evaluated the physicochemical and morphological properties of a marine sponge protein extract (PE) using scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDS), analysis of mass loss and pH and in vitro and in vivo. Scanning electron microscopy showed that PE fibers present a granular aspect and irregular structure and the element carbon followed by oxygen was detected in the EDS analysis. Moreover, a 29% of mass loss was observed after 14 days and the pH slightly modified after 14 days. Cell viability of fibroblast cells (L929) of control and PE at a concentration of 25% demonstrated higher values compared to the groups. Osteoblast cell viability of PE at 25 and 50% was significantly higher. Comet assay on day 1 showed higher values for PE at 25%. In addition, in vivo experiments demonstrated that in the treated animals, the bone defects were filled with biomaterial particles, granulation tissue and some areas of newly formed bone. Furthermore, similar immunoexpression of Runx-2 and Cox-2 was observed. Taken together, all results suggest that PE is biocompatible, present non-citotoxicity in the in vitro studies (at the lower concentration) and in the in vivo studies and it can be considered as an alternative source of collagen for tissue engineering proposals.

In vitro and in vivo genotoxicity and cytotoxicity analysis of protein extract from Aplysina fulva sponges

This study evaluated the physicochemical and morphological properties of a marine sponge protein extract (PE) using scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDS), analysis of mass loss and pH and in vitro and in vivo. Scanning electron microscopy showed that PE fibers present a granular aspect and irregular structure and the element carbon followed by oxygen was detected in the EDS analysis. Moreover, a 29% of mass loss was observed after 14 days and the pH slightly modified after 14 days. Cell viability of fibroblast cells (L929) of control and PE at a concentration of 25% demonstrated higher values compared to the groups. Osteoblast cell viability of PE at 25 and 50% was significantly higher. Comet assay on day 1 showed higher values for PE at 25%. In addition, in vivo experiments demonstrated that in the treated animals, the bone defects were filled with biomaterial particles, granulation tissue and some areas of newly formed bone. Furthermore, similar immunoexpression of Runx-2 and Cox-2 was observed. Taken together, all results suggest that PE is biocompatible, present non-citotoxicity in the in vitro studies (at the lower concentration) and in the in vivo studies and it can be considered as an alternative source of collagen for tissue engineering proposals.(AU)

Cytotoxicity assessment of beta-lapachone in endothelial cells

The selective activity of an antineoplastic drug is related to its ability to promote cytotoxic action on tumor cells and preserve the integrity of non-neoplastic cells. Beta-lapachone is extracted from the sawdust of Ipe wood, a thick bark tree from the Ipe wood found in the Brazilian Cerrado biome. This study aimed to evaluate the cytotoxic action of beta-lapachone in an endothelial cell line. The EA.hy926 cells were seeded in two groups, G1 and G2, cultured and exposed to beta-lapachone at concentrations of 0.0, 0.01, 0.03, 0.1, 0.3, 1 and 3 & 956;M for 24 hours. G1 remained under normal cultivation conditions and G2 was subjected to oxidative stress through an ischemia and reperfusion assay, in a deoxygenated sealed chamber. The cytotoxicity assay was performed using the tetrazolium reduction method. In G1, the cytotoxicity ranged from 0.0 to 10.0%; and in G2 between 0.0 and 6.3%. No statistically significant difference was observed between the obtained values. Moreover, we found no cytotoxic action of beta-lapachone on endothelial cells, and the results point out that the drug might have preserved the cell’s integrity against oxidative stress under the conditions of this experiment. This promising result suggests the possibility of beta-lapachone as a chemotherapy drug with selective activity.

Cytotoxicity assessment of beta-lapachone in endothelial cells

The selective activity of an antineoplastic drug is related to its ability to promote cytotoxic action on tumor cells and preserve the integrity of non-neoplastic cells. Beta-lapachone is extracted from the sawdust of Ipe wood, a thick bark tree from the Ipe wood found in the Brazilian Cerrado biome. This study aimed to evaluate the cytotoxic action of beta-lapachone in an endothelial cell line. The EA.hy926 cells were seeded in two groups, G1 and G2, cultured and exposed to beta-lapachone at concentrations of 0.0, 0.01, 0.03, 0.1, 0.3, 1 and 3 & 956;M for 24 hours. G1 remained under normal cultivation conditions and G2 was subjected to oxidative stress through an ischemia and reperfusion assay, in a deoxygenated sealed chamber. The cytotoxicity assay was performed using the tetrazolium reduction method. In G1, the cytotoxicity ranged from 0.0 to 10.0%; and in G2 between 0.0 and 6.3%. No statistically significant difference was observed between the obtained values. Moreover, we found no cytotoxic action of beta-lapachone on endothelial cells, and the results point out that the drug might have preserved the cells integrity against oxidative stress under the conditions of this experiment. This promising result suggests the possibility of beta-lapachone as a chemotherapy drug with selective activity.(AU)

Hematological responses to hyperimmune plasma low dose administration in foals during weaning / Respostas hematológicas da administração de baixa dose de plasma hiperimune em potros no período de desmame

Weaning is a development stage that brings maternal independence, change of management and feeding, leading to immune deficit. The objective of this study was to evaluate the hematological and biochemical parameters of foals in weaning phase after intravenous administration of hyperimmune plasma (1 mL/Kg) in low dose. Ten foal weaned from the Mangalarga Machador breed were used, being divided in control (GC; n=5) and treated group (GT; n=5). The results obtained show that hematological and biochemical parameters of foals that received low doses of hyperimmune plasma were within the reference range for the equine species.

O desmame é uma fase do desenvolvimento que traz independência materna, mudança de manejo e alimentação, levando ao déficit imunológico. O objetivo deste estudo foi avaliar os parâmetros hematológicos e bioquímicos de potros em fase de desmame após administração intravenosa de plasma hiperimune (1 mL / Kg) em baixa dose. Foram utilizados dez potros des-mamados da raça Mangalarga Machador, sendo divididos em grupo controle (GC; n = 5) e grupo tratado (GT; n = 5). Os resul-tados obtidos mostram que os parâmetros hematológicos e bioquímicos de potros que receberam baixas doses de plasma hiperi-mune estavam dentro da faixa de referência para a espécie equina.

Hematological responses to hyperimmune plasma low dose administration in foals during weaning / Respostas hematológicas da administração de baixa dose de plasma hiperimune em potros no período de desmame

Weaning is a development stage that brings maternal independence, change of management and feeding, leading to immune deficit. The objective of this study was to evaluate the hematological and biochemical parameters of foals in weaning phase after intravenous administration of hyperimmune plasma (1 mL/Kg) in low dose. Ten foal weaned from the Mangalarga Machador breed were used, being divided in control (GC; n=5) and treated group (GT; n=5). The results obtained show that hematological and biochemical parameters of foals that received low doses of hyperimmune plasma were within the reference range for the equine species.(AU)

O desmame é uma fase do desenvolvimento que traz independência materna, mudança de manejo e alimentação, levando ao déficit imunológico. O objetivo deste estudo foi avaliar os parâmetros hematológicos e bioquímicos de potros em fase de desmame após administração intravenosa de plasma hiperimune (1 mL / Kg) em baixa dose. Foram utilizados dez potros des-mamados da raça Mangalarga Machador, sendo divididos em grupo controle (GC; n = 5) e grupo tratado (GT; n = 5). Os resul-tados obtidos mostram que os parâmetros hematológicos e bioquímicos de potros que receberam baixas doses de plasma hiperi-mune estavam dentro da faixa de referência para a espécie equina.(AU)

Hematologic response and serum and peritoneal fluid proteinogram of cattle affected by intestinal diseases and traumatic reticuloperitonitis / Resposta hematológica e proteinograma sérico e do líquido peritoneal de bovinos acometidos com enfermidades intestinais e reticuloperitonite traumática

This study aimed to evaluate the hematologic response and the serum and peritoneal fluid (PF) proteinogram of cattle affected by digestive diseases. Twenty-seven animals were distributed in two groups: GI (intestinal diseases) and GII (traumatic reticuloperitonitis, TRP). The animals were previously submitted to a physical exam. Subsequently, blood samples were collected to perform the complete blood count, determine the plasma protein and fibrinogen, and obtain the serum for proteinogram in polyacrylamide gel (SDS-PAGE). Simultaneously, PF was collected to perform physical and chemical evaluation and the electrophoretic profile (SDS-PAGE). ANOVA at the 5% probability level was used to compare the groups. The animals showed signs of apathy, dehydration, and gastrointestinal hypomotility in both groups. However, GI animals showed more significant clinical changes. The blood count of both groups (P > 0.05) showed leukocytosis due to neutrophilia and a regenerative left shift with hyperfibrinogenemia. The proteinogram of both body fluids allowed the identification of proteins albumin (ALB), transferrin (TRF), ceruloplasmin, haptoglobin, α1-acid glycoprotein (α1-AGP), MW 23000 Da, α1-antitrypsin, IgA, and IgG. The [PT] PF/[PT] blood serum ratio of each of the identified proteins increased, showing statistical differences between groups (P < 0.05) regarding PT, ALB, TRF, α1-AGP, and IgG values, with GI animals showing the highest ratio. Intestinal diseases and TRP triggered a systemic and local response characterized by clinical, hematological, and serum and PF proteinogram alterations. The proteins α1-GPA, haptoglobin, and TRF measured in PF were good inflammation biomarkers and useful as an auxiliary tool for the diagnosis and prognosis of digestive diseases in cattle.(AU)

O objetivo deste estudo foi avaliar a resposta hematológica e o proteinograma sérico e do líquido peritoneal (LP) de bovinos acometidos com doenças digestivas. Foram avaliados 27 bovinos distribuídos em dois grupos, GI (enfermidades intestinais) e GII (reticuloperitonite traumática-RPT). Os animais foram previamente submetidos ao exame físico. Posteriormente foram colhidas amostras de sangue para realização do hemograma, determinação plasmática da proteína e do fibrinogênio e obtenção do soro para realização do proteinograma em gel de poliacrilamida (SDS-PAGE). Simultaneamente foi colhido o LP para avaliação física e química, assim como a realização do perfil eletroforético (SDS-PAGE). Empregou-se a análise de variância ao nível de 5% de probabilidade visando à comparação entre os grupos. Em ambos os grupos os animais demonstraram sinais de apatia, desidratação e hipomotilidade gastrointestinal, no entanto, os animais do GI apresentaram alterações clínicas mais expressivas. No hemograma observou-se em ambos os grupos (P > 0,05) leucocitose por neutrofilia e desvio à esquerda regenerativo com hiperfibrinogenia. O proteinograma de ambos os fluidos corpóreos permitiu a identificação das proteínas albumina (ALB), transferrina (TRF), ceruloplasmina, haptoglobina, α-1 glicoproteína ácida (α1-GPA), PM 23.000 Da, α-1 anti-tripsina, IgA e IgG. Os valores da relação [PT] LP / [PT] soro sanguíneo de cada uma das proteínas identificadas demonstrou elevação dos mesmos, bem como diferença estatística entre grupos (P < 0,05) nos valores da PT, ALB, TRF, α1-GPA e IgG, nos quais a relação foi mais elevada nos animais do GI. As enfermidades intestinais e a RPT desencadearam resposta sistêmica e local caracterizada pelas alterações clínicas, hematológicas, e do proteinograma sérico e do LP. A α1-GPA, a haptoglobina e a TRF, mensuradas no LP se mostraram bons biomarcadores de inflamação, sendo úteis como recurso auxiliar de diagnóstico e prognóstico das doenças digestivas dos bovinos.(AU)

Clinical, hematological and risk factors associated with borreliosis in horses from Sao Paulo, Brazil / Fatores clínicos, hematológicos e de risco associados à borreliose em cavalos de São Paulo, Brasil

Lyme borreliosis is caused by Borrelia burgdorferi sensu lato and affects humans and many other mammals, including horses. This disease is poorly studied and reported in horses, and epidemiological surveys are required to provide more precise information about the course of the disease. The aims of the present study were to determine the prevalence of seropositive horses for Borrelia burgdorferi sensu lato in São Paulo State, Brazil and to collect data on possible risk factors associated with the disease along with clinical and hematological changes in seropositive horses. There was verified that there was a high correlation between the occurrence of seropositive horses infested with Amblyomma sculptum ticks and the presence of capybaras on the property as well as the occurrence of abortion and retained placenta in mares. In terms of hematological alterations, the occurrence of lymphopenia was observed in seropositive animals. Borreliosis in horses from São Paulo, Brazil can be associated with presence of Amblyomma scupltum ticks, proximity with capybaras and can be manifested as alterations in reproduction of mares and lymphopenia.

A borreliose de Lyme é causada por Borrelia burgdorferi sensu lato e afeta humanos e muitos outros mamíferos, incluindo cavalos. Esta doença é pouco estudada e relatada em equinos, sendo necessários levantamentos epidemiológicos para fornecer informações mais precisas sobre o curso da doença. Os objetivos do presente estudo foram determinar a prevalência de cavalos soropositivos para Borrelia burgdorferi sensu lato no Estado de São Paulo, Brasil e coletar dados sobre possíveis fatores de risco associados à doença, juntamente com alterações clínicas e hematológicas em cavalos soropositivos. Verificou-se que houve alta correlação entre a ocorrência de equinos soropositivos infestados por carrapatos Amblyomma sculptum e a presença de capivaras nas propriedades, bem como a ocorrência de abortamento e retenção de placenta em éguas. Em termos de alterações hematológicas, observou-se a ocorrência de linfopenia em animais soropositivos. A borreliose em cavalos de São Paulo, Brasil, pode estar associada à presença de carrapatos Amblyomma scupltum, proximidade com capivaras e se manifestar clinicamente como alterações reprodutivas em éguas e linfopenia.

Clinical, hematological and risk factors associated with borreliosis in horses from Sao Paulo, Brazil / Fatores clínicos, hematológicos e de risco associados à borreliose em cavalos de São Paulo, Brasil

Lyme borreliosis is caused by Borrelia burgdorferi sensu lato and affects humans and many other mammals, including horses. This disease is poorly studied and reported in horses, and epidemiological surveys are required to provide more precise information about the course of the disease. The aims of the present study were to determine the prevalence of seropositive horses for Borrelia burgdorferi sensu lato in São Paulo State, Brazil and to collect data on possible risk factors associated with the disease along with clinical and hematological changes in seropositive horses. There was verified that there was a high correlation between the occurrence of seropositive horses infested with Amblyomma sculptum ticks and the presence of capybaras on the property as well as the occurrence of abortion and retained placenta in mares. In terms of hematological alterations, the occurrence of lymphopenia was observed in seropositive animals. Borreliosis in horses from São Paulo, Brazil can be associated with presence of Amblyomma scupltum ticks, proximity with capybaras and can be manifested as alterations in reproduction of mares and lymphopenia.(AU)

A borreliose de Lyme é causada por Borrelia burgdorferi sensu lato e afeta humanos e muitos outros mamíferos, incluindo cavalos. Esta doença é pouco estudada e relatada em equinos, sendo necessários levantamentos epidemiológicos para fornecer informações mais precisas sobre o curso da doença. Os objetivos do presente estudo foram determinar a prevalência de cavalos soropositivos para Borrelia burgdorferi sensu lato no Estado de São Paulo, Brasil e coletar dados sobre possíveis fatores de risco associados à doença, juntamente com alterações clínicas e hematológicas em cavalos soropositivos. Verificou-se que houve alta correlação entre a ocorrência de equinos soropositivos infestados por carrapatos Amblyomma sculptum e a presença de capivaras nas propriedades, bem como a ocorrência de abortamento e retenção de placenta em éguas. Em termos de alterações hematológicas, observou-se a ocorrência de linfopenia em animais soropositivos. A borreliose em cavalos de São Paulo, Brasil, pode estar associada à presença de carrapatos Amblyomma scupltum, proximidade com capivaras e se manifestar clinicamente como alterações reprodutivas em éguas e linfopenia.(AU)

Characterization of proteinuria in treated and untreated dogs naturally infected with Leishmania sp / Caracterização da proteinúria de cães tratados e não tratados, naturalmente infectados com Leishmania sp

In the search for an early biomarker of renal injury, this study aimed to determine the urinary protein profile of dogs with leishmaniasis without treatment and treated as determined by Brazilian legislation. The identification of proteinuria, its classification and the circumstances in which it takes place instigated this study. For this, 30 dogs from an outpatient clinic at a Veterinary Hospital in Belo Horizonte were evaluated. All animals underwent clinical and laboratory tests, which included renal biomarkers. The proteins were characterized using the SDS-page electrophoresis technique, and thus, a urinary protein profile was developed comparing patients considered clinically healthy with dogs infected with leishmaniasis that were under treatment and with untreated infected dogs. The results showed that the hematological and biochemical parameters showed similar behavior between the groups of healthy dogs and dogs with leishmaniasis treated, however a very heterogeneous pattern of urinary proteins can be observed and differed between healthy animals and animals with leishmaniasis, as well as between treated and untreated animals. The results suggest that the classification of proteinuria can be a tool that helps in the staging of animals infected with L. infantum and can differentiate them as to the severity of existing kidney injuries.(AU)

Na busca por um biomarcador precoce de injúria renal, este trabalho teve como objetivo determinar o perfil proteico urinário de cães infectados com leishmaniose sem tratamento e tratados conforme determina a legislação brasileira. A identificação da proteinúria, sua classificação e as circunstâncias em que ocorrem instigaram este estudo. Para tanto, foram avaliados 30 cães oriundos do atendimento clínico ambulatorial de um Hospital Veterinário em Belo Horizonte. Todos os animais passaram por exame clínico e laboratorial, que incluíram biomarcadores renais. As proteínas foram caracterizadas através da técnica de eletroforese por SDS-PAGE, e assim, foi elaborado um perfil proteico urinário comparando pacientes considerados clinicamente hígidos, com cães infectados por Leishmania (L.) infantum e que estavam sob tratamento e cães infectados não tratados. Os resultados demonstraram que os parâmetros hematológicos e bioquímicos apresentaram comportamento semelhante entre os grupos de cães hígidos e de cães infectados com L. infantum tratados, entretanto um padrão muito heterogêneo de proteínas urinárias pode ser observado e diferiu entre animais hígidos e animais com leishmaniose, assim como entre os animais tratados e não tratados. Os resultados sugerem que a classificação da proteinúria pode ser uma ferramenta que auxilia no estadiamento de animais infectados por L. infantum podendo diferenciá-los quanto à gravidade de lesões renais existentes.(AU)