Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.(AU)

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.(AU)

Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

Molecular, serological, and parasitological detection of Babesia vogeli in dogs in the state of Piauí, Brazil / Detecção molecular, sorológica e parasitológica de Babesia vogeli em cães do estado do Piauí, Brasil

Studies on canine babesiosis in northeastern Brazil are scarce, although the weather conditions in this region are favorable for the development of the tick vector. This study determined the prevalence of Babesia vogeli in dogs sampled in Teresina, state of Piauí, northeast Brazil, using direct and indirect diagnostic methods and performed a phylogenetic analysis of 18S rRNA sequences. A total of 315 dogs were screened during routine care regardless of clinical suspicion. Blood was collected by jugular venipuncture to perform indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR) and for parasite screening in peripheral blood smears. Positivity was 2.2% (7/315) by microscopy, 4.8% (15/315) by PCR, and 48.6% (153/315) by IFA. PCR amplified a 602-bp fragment of the piroplasmid 18S rRNA gene, and sequence alignment and analysis revealed 99% homology with B. vogeli isolates from other regions of Brazil and other countries. In addition, there was high variability among sequences from other northeast states of Brazil. This study is the first to perform the molecular analysis of B. vogeli in Piauí. The results demonstrate that canine babesiosis is endemic in dogs sampled in Teresina and that PCR may be the method of choice to perform parasite screening in this region.

Estudos sobre a babesiose canina são escassos no Nordeste do Brasil, apesar das condições climáticas favoráveis ao desenvolvimento do carrapato vetor. Esta pesquisa objetivou determinar a ocorrência de Babesia vogeli em cães amostrados em Teresina, estado do Piauí, região Meio Norte do Brasil, através de métodos diretos e indiretos de diagnóstico, além de realizar análise filogenética das sequências 18S rRNA de piroplasmídeos obtidas no estudo. Foram avaliados 315 cães atendidos em clínicas veterinárias, sob qualquer suspeita clínica. Desses animais, foi colhido sangue por venopunção jugular para Reação de Imunofluorescência Indireta (RIFI) e Reação em Cadeia pela Polimerase (PCR). Além disso, esfregaços de sangue periférico foram realizados para pesquisa direta do parasita. A positividade dos animais foi de 2,2% (7/315) ao esfregaço sanguíneo, 4,8% (15/315) à PCR e 48,6% (153/315) à RIFI. O sequenciamento de amostras positivas à PCR resultou em um fragmento de 602 pb do gene 18S rRNA de piroplasmídeos, cujo alinhamento e análise da sequência revelaram 99% de homologia com isolados de B. vogeli de outras regiões do Brasil, além de outros países. É interessante ressaltar que, comparando isolados em diferentes estados do Nordeste, a homologia pode ser bastante variável. Esses são os primeiros resultados sobre a análise molecular de B. vogeli no Estado do Piauí. Além disso, este estudo demonstra que a babesiose canina é endêmica em cães de Teresina, Nordeste do Brasil, e que a PCR pode ser o método de escolha para diagnóstico da doença nessas áreas.

Molecular, serological, and parasitological detection of Babesia vogeli in dogs in the state of Piauí, Brazil / Detecção molecular, sorológica e parasitológica de Babesia vogeli em cães do estado do Piauí, Brasil

Studies on canine babesiosis in northeastern Brazil are scarce, although the weather conditions in this region are favorable for the development of the tick vector. This study determined the prevalence of Babesia vogeli in dogs sampled in Teresina, state of Piauí, northeast Brazil, using direct and indirect diagnostic methods and performed a phylogenetic analysis of 18S rRNA sequences. A total of 315 dogs were screened during routine care regardless of clinical suspicion. Blood was collected by jugular venipuncture to perform indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR) and for parasite screening in peripheral blood smears. Positivity was 2.2% (7/315) by microscopy, 4.8% (15/315) by PCR, and 48.6% (153/315) by IFA. PCR amplified a 602-bp fragment of the piroplasmid 18S rRNA gene, and sequence alignment and analysis revealed 99% homology with B. vogeli isolates from other regions of Brazil and other countries. In addition, there was high variability among sequences from other northeast states of Brazil. This study is the first to perform the molecular analysis of B. vogeli in Piauí. The results demonstrate that canine babesiosis is endemic in dogs sampled in Teresina and that PCR may be the method of choice to perform parasite screening in this region.(AU)

Estudos sobre a babesiose canina são escassos no Nordeste do Brasil, apesar das condições climáticas favoráveis ao desenvolvimento do carrapato vetor. Esta pesquisa objetivou determinar a ocorrência de Babesia vogeli em cães amostrados em Teresina, estado do Piauí, região Meio Norte do Brasil, através de métodos diretos e indiretos de diagnóstico, além de realizar análise filogenética das sequências 18S rRNA de piroplasmídeos obtidas no estudo. Foram avaliados 315 cães atendidos em clínicas veterinárias, sob qualquer suspeita clínica. Desses animais, foi colhido sangue por venopunção jugular para Reação de Imunofluorescência Indireta (RIFI) e Reação em Cadeia pela Polimerase (PCR). Além disso, esfregaços de sangue periférico foram realizados para pesquisa direta do parasita. A positividade dos animais foi de 2,2% (7/315) ao esfregaço sanguíneo, 4,8% (15/315) à PCR e 48,6% (153/315) à RIFI. O sequenciamento de amostras positivas à PCR resultou em um fragmento de 602 pb do gene 18S rRNA de piroplasmídeos, cujo alinhamento e análise da sequência revelaram 99% de homologia com isolados de B. vogeli de outras regiões do Brasil, além de outros países. É interessante ressaltar que, comparando isolados em diferentes estados do Nordeste, a homologia pode ser bastante variável. Esses são os primeiros resultados sobre a análise molecular de B. vogeli no Estado do Piauí. Além disso, este estudo demonstra que a babesiose canina é endêmica em cães de Teresina, Nordeste do Brasil, e que a PCR pode ser o método de escolha para diagnóstico da doença nessas áreas.(AU)

Classification of pulmonary alterations in canine visceral leishmaniasis / Classificação das alterações pulmonares na leishmaniose visceral canina

The aim of this study was to identify and classify the pulmonary alterations in dogs naturally infected by Leishmania (Leishmania) infantum according to clinical manifestation. After diagnosis to VL by parasitological exam, 24 infected dogs were subdivided in two groups: 10 dogs with until four clinical signs; and 14 dogs with five or more clinical signs. As control were evaluated 5 healthy dogs. Pulmonary tissue samples were stained by hematoxylin-eosin (H-E) and Masson's Trichrome to histopathological examination, and Grocotts methenamine silver to exclude fungal infections, as well immunoperoxidase to detection of Leishmania. Histopathological examination of the lungs showed a focal thickening of the alveolar septa due to congestion, edema and cellular infiltrate. Moreover, presence of fibrosis, bronchiolar exudate and flaking of the bronchiolar epithelium, presence of hemosiderin and hyperplasia of goblet cell were observed. The pattern fibrotic-cellular lesions predominated in most infected dogs but mainly in infected dogs with five or more signs clinical. However, fibrosis was higher in infected dogs with four signs clinical. Immunoperoxidase staining showed amastigotes and/or antigen of Leishmania in all infected dogs but there was an increase significant in infected dogs with five or more signs clinical. Semi-quantitative analysis of intensities antigen reveled a positive correlation between fibrosis in infected dogs. In conclusion, the presence of the parasite and/or antigens modifies the lung architecture due for fibrotic-cellular pattern in infected dogs with severe manifestation clinical.(AU)

O objetivo desse estudo foi identificar e classificar as alterações pulmonares de cães naturalmente infectados por Leishmania (Leishmania) infantum conforme as manifestações clínicas. Após o diagnóstico para LV pelo exame parasitológico, 24 cães infectados foram subdivididos em dois grupos: 10 cães com até quatro sinais clínicos; e 14 cães com cinco ou mais sinais clínicos. Como controle foram avaliados 5 cães sadios. Amostras de tecido pulmonar foram coradas com hematoxilina-eosina (H-E) e Tricrômico de Masson para avaliação histopatológica, e prata-metenamina de Grocott para excluir infecções fúngicas, bem como imunoperoxidase para detecção de Leishmania. A análise microscópica do pulmão revelou espessamento focal do septo interalveolar devido à congestão, edema e infiltrado celular. Além disso, presença de fibrose, exsudato bronquiolar e ainda descamação do epitélio bronquiolar com perda de cílios, presença de hemossiderina e hiperplasia de células caliciformes. O padrão de lesão fibrótico-celular predominou nos cães infectados com cinco ou mais sinais clínicos. Em contrapartida, fibrose estava presente em intensidade maior no grupo nos cães infectados com até quatro sinais clínicos. A análise imunoistoquímica mostrou maior intensidade de antígeno e amastigotas de Leishmania nos cães infectados com cinco ou mais sinais clínicos. A análise semi-quantitativa do antígeno mostrou correlação positiva de acordo com a intensidade da fibrose. Concluiu-se que a presença do parasita e seus antígenos modificam a arquitetura pulmonar evoluindo, em sua maioria, para o padrão fibrótico-celular em associação à intensidade das manifestações clínicas.(AU)

Detecção de Brucella ovis por reação em cadeia da polimerase (PCR) em amostras de urina de ovinos naturalmente infectados na microrregião de Teresina, PI / Brucella ovis detection by polymerase chain reaction (PCR) in naturally infected sheep urine samples in the micro region of Teresina, PI

Ovine Brucellosis is an infectious disease responsible for the production of reproductive disorders,triggering economic losses to sheep production. The researchs objective was detect B. ovis DNA in sheepsbelonging to cities in the micro region of Teresina-PI. Thus, were collected 100 urine and blood samples ofsheeps older than or equal to six months. Urine samples were subjected to conventional PCR and samples ofblood AGID technique. From 100 blood samples, 17 (17%) were reactive to AGID. In conventional PCR, from100 urine samples, six (6%) were positive. There was agreement between the tests, however, was consideredweak (18.9%), based on the Kappa coefficient ranking. Thus, it was observed that both techniques are valid and can be regarded as tools of choice for the diagnosis of ovine brucellosis.(AU)

Detecção de Brucella ovis por reação em cadeia da polimerase (PCR) em amostras de urina de ovinos naturalmente infectados na microrregião de Teresina, PI / Brucella ovis detection by polymerase chain reaction (PCR) in naturally infected sheep urine samples in the micro region of Teresina, PI

Ovine Brucellosis is an infectious disease responsible for the production of reproductive disorders,triggering economic losses to sheep production. The research’s objective was detect B. ovis DNA in sheepsbelonging to cities in the micro region of Teresina-PI. Thus, were collected 100 urine and blood samples ofsheeps older than or equal to six months. Urine samples were subjected to conventional PCR and samples ofblood AGID technique. From 100 blood samples, 17 (17%) were reactive to AGID. In conventional PCR, from100 urine samples, six (6%) were positive. There was agreement between the tests, however, was consideredweak (18.9%), based on the Kappa coefficient ranking. Thus, it was observed that both techniques are valid and can be regarded as tools of choice for the diagnosis of ovine brucellosis.

Transmissible venereal tumor treated with autohemotherapy

Background: Transmissible venereal tumor (TVT) is a neoplasm of round cells, that affects exclusively the canine specie. The etiology of the tumor is unknown, but the hystiocitic hipothesis is the more accepted. It is transmitted principally by the venereal form, but the inplant of cells can cause the disease. The clinic signs of the neoplasia are typical and include tumoral friable mass (in shape of cauliflower), that bleed easily. The most common localization is external genitalia, but the tumor can appears in skin surface and other organs. TVT grows rapidly (progressive phase) after transplantation, followed by a static phase (without cell proliferation) and then may regress spontaneously (stage of regression). The regression of the neoplasia is associated with increased infi ltration of T lymphocytes and macrophages in the tumor and characterized by increased apoptosis of tumor cells and fi brosis. The diagnosis of TVT is usually done by the physical aspect of the tumor, and confirmed by cytology or histopathology. TVT can be prevented by castration of the animals. The treatment of TVT is usually performed with vincristine, which has side effects, requires care in its application, and has a relatively high cost. Thus, new therapeutic low-cost alternatives are suggested, as is the case of autohemotherapy. The autohemotherapy technique consists of administration of autologous whole blood intramuscularly. In veterinary practice has been used successfully to treat bovine papillomatosis and other diseases. Probably the mechanism of action of autohemotherapy is to enhance organic immunity. This report aimed to evaluate the behavior of the transmissible venereal tumor in six dogs with naturally transplanted tumor after treatment with autohemotherapy. Materials, Methods & Results: Six adult bitches were clinically examined and all presented transmissible venereal tumor located in external genitalia, adquired by natural transmission. Cytological examination confirmed the diagnosis of TVT. The bitches were kept in kennels by one week before treatment, and it was not observed natural regression of tumor mass. After this time the animals were submitted to treatment with autohemotherapy, which consisted of application of autologous whole blood in the gluteal muscles, at dose of 10 mL. The applications were made weekly for seven weeks. All tumors were measured before and after autohemotherapy and, also weekly during the treatment. All tumors were friable and bleeding easily, multilobulated, nodulares, cauliflower-like shape, pale-pink to pale red, with surface smooth or irregular. In some animals there were signs of secondary infection. In general, the measurement of tumor ranged from 3.0 cm to 7.1 cm before of autohemotherapy. After treatment it was observed, macroscopically, a decrease of the tumoral mass in three dogs. Discussion: It was established as criterion for stopping treatment the time of seven weeks. Possibly if the treatment had been extended the regression could have been completed. Probably autohemotherapy increased immunity and, consequently, contributed to increase the body's resistance against TVT, producing regression of tumoral mass. Therefore, the autohemotherapy led to macroscopic partial regression of the tumor in 50% of animals subjected to this treatment, stimulating further research in this area.