Canine distemper virus detection in asymptomatic and non vaccinated dogs

A quantitative real time polymerase chain reaction (PCR) revealed canine distemper virus presence in peripheral blood samples from asymptomatic and non vaccinated dogs. Samples from eleven domestic dogs with no signs of canine distemper and not vaccinated at the month of collection were used. Canine distemper virus vaccine samples in VERO cells were used as positive controls. RNA was isolated with Trizol®, and treated with a TURBO DNA-free kit. Primers were designed for canine distemper virus nucleocapsid protein coding region fragment amplification (84 bp). Canine b-actin (93 bp) was utilized as the endogenous control for normalization. Quantitative results of real time PCR generated by ABI Prism 7000 SDS Software showed that 54.5% of dogs with asymptomatic canine distemper were positive for canine distemper virus. Dissociation curves confirmed the specificity of the real time PCR fragments. This technique could detect even a few copies of viral RNA and identificate subclinically infected dogs providing accurate diagnosis of this disease at an early stage.

Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO², >37ºC). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

Doenças da cavidade nasal em ruminantes no Brasil

Este trabalho descreve as doenças das fossas nasais diagnosticadas em ruminantes no Hospital Veterinário da Universidade Federal de Campina Grande, em Patos, Paraíba, nos anos de 2003-2009. No período foram registrados três diagnósticos de doenças das fossas nasais de bovinos, três em caprinos e nove em ovinos (de um total de 404 diagnósticos em bovinos, 330 em caprinos e 338 em ovinos). Descrevem-se um caso de rinite atópica em bovinos, sete surtos de conidiobolomicose e dois de pitiose rinofacial em ovinos, dois casos de prototecose e um de aspergilose nasal em caprinos e um mixoma e um fibrossarcoma em bovinos. Adicionalmente, é realizada uma revisão de outras doenças das fossas nasais de ruminantes descritas em outras regiões do Brasil, incluindo oestrose, rinosporidiose, carcinoma epidermóide e tumor etmoidal enzoótico.

Frequencies of DEA blood types in a purebred canine blood donor population in Porto Alegre, RS, Brazil

The study of canine immunohematology is very important for veterinary transfusion medicine. The objective of this study was to determine the DEA blood type frequencies in a purebred canine blood donor population from Porto Alegre, RS, Brazil. One hundred clinically healthy purebred dogs were chosen, 20 dogs from each breed (Great Dane, Rottweiler, Golden Retriever, German Shepherd and Argentine Dogo). Blood samples were taken in ACD-A tubes and the MSU hemagglutination tube test (MI, USA) was used to determine the blood types. The studied population presented general frequencies of 61% for DEA 1.1, 22% for DEA 1.2, 7% for DEA 3, 100% for DEA 4, 9% for DEA 5 and 16% for DEA 7. A significant association was found between breeds and certain combinations of blood types in this population. The results are in agreement with the literature since most part of the canine population studied was positive for DEA 1.1, the most antigenic blood type in dogs. Differences were found among the studied breeds and those should be considered when selecting a blood donor. The knowledge of blood types frequencies and their combinations in different canine populations, including different breeds, is important because it shows the particularities of each group, helps to keep a data bank of local frequencies and minimizes the risks of transfusion reactions.

Canine papillomatosis: a retrospective study of 24 cases (2001-2011) and immunohistochemical characterization

A retrospective study of 24 cases of papillomas in dogs was performed from January 2001 to March 2011. Additionally, immunohistochemistry (IHC) was used to characterize and evaluate the samples. We found that disease was observed more in mixed breed dogs, ages ranging from 6 months to 10 years (mean 3.1 years), and there was no gender predilection. The main lesion sites were the skin (75%), lips (16.7%), and eyelids (8.3%). Upon histological evaluation, we observed papillary exophytic proliferation of squamous epithelium and papillary endophytic proliferation (inverted) in 87.5% and 12.5% of cases, respectively. The tumors were characterized by spinous layer hyperplasia (87.5%) with koilocytes (70.8%) and intranuclear pale basophilic inclusions bodies (8.3%), prominent granular layer with large amounts of keratohyalin granules (95.8%), and hyperkeratosis in the stratum corneum (100%). Positive immunostaining for Papillomavirus was found in 83.3% of cases, which were distributed between the granular layer and the stratum corneum. These findings indicate the following: that papillomas in dogs are caused by Papillomavirus, the viral cytopathic effect induces epithelial lesions, viral particles are found inside the cell nuclei, and inclusions bodies are rare.

A phylogenetic study of canine parvovirus type 2c in midwestern Brazil

Since the late 1970s, canine parvovirus type 2 (CPV-2) has emerged as a causative agent of fatal severe acute hemorrhagic enteritis in dogs. To date, three antigenic types of CPV-2 were described worldwide (CPV-2a/b/c). This study was conducted to determine the variants of CPV-2 circulating in dogs from the Cuiabá Municipality in Midwestern Brazil. Out of 50 fecal samples, collected between 2009 and 2011, 27 tested positive for CPV-2. A 583 bp fragment of the VP2 gene was amplified by PCR, 13 representative samples were analyzed further by DNA sequencing. All strains were characterized as CPV-2c, displayed a low genetic variability although observed several amino acid substitution. These findings indicated that CPV-2c has been circulating in dogs from the Cuiabá Municipality in Midwestern Brazil.

Canine distemper virus infection in a lesser grison (Galictis cuja): first report and virus phylogeny

Infectious diseases in wild animals have been increasing as a result of their habitat alterations and closer contact with domestic animals. Canine distemper virus (CDV) has been reported in several species of wild carnivores, presenting a threat to wildlife conservation. We described the first case of canine distemper virus infection in lesser grison (Galictis cuja). A free-ranging individual, with no visible clinical sigs, presented sudden death after one day in captivity. Molecular diagnosis for CDV infection was performed using whole blood collected by postmortem intracardiac puncture, which resulted positive. The virus phylogeny indicated that domestic dogs were the probable source of infection.

Outbreak of autochthonous canine visceral leishmaniasis in Santa Catarina, Brazil

The present study reports the first outbreak of autochthonous canine visceral leishmaniasis in Florianópolis, Santa Catarina, southern Brazil. Following the report of two cases of CVL, the Control Center of Zoonotic Diseases conducted a serological survey by ELISA and IFAT assays in seven districts of the Santa Catarina Island. Eleven seropositive dogs of autochthonous transmission were used in the present study. Infection by Leishmania sp. was confirmed by parasitological examination of bone marrow, liver, spleen and lymph nodes, culture in Schneider's medium and PCR. Leishmania sp. isolates were characterized by PCR-RFLP and hybridization with specific probes, allowing for the identification of Leishmania infantum. Autochthonous transmission of this disease in an area with high tourist traffic presents a major public health concern and signifies the emergence of an important zoonosis in southern Brazil. Therefore, the implementation of surveillance and control measures is imperative to prevent the spread of the disease among the canine population as well as transmission to the human population.

Expression of P-glycoprotein, multidrug resistance-associated protein, glutathione-S-transferase pi and p53 in canine transmissible venereal tumor

The overexpression of proteins P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP1), mutant p53, and the enzyme glutathione-S-transferase (GSTpi) are related to resistance to chemotherapy in neoplasms. This study evaluated the expression of these markers by immunohistochemistry in two groups of canine TVT, without history of prior chemotherapy (TVT1, n=9) and in TVTs presented unsatisfactory clinical response to vincristine sulfate (TVT2, n=5). The percentage of specimens positively stained for P-gp, MRP1, GSTpi and p53 were, respectively 88.8%, 0%, 44.5% and 22.2% in TVT1 and 80%, 0%, 80% and 0% in TVT2. In TVT1, one specimen presented positive expression for three markers and four specimens for two markers. In TVT2, three specimens expressed P-gp and GSTpi. In conclusion, the canine TVTs studied expressed the four markers evaluated, but just P-gp and GSTpi were significantly expressed, mainly at cytoplasm and cytoplasm and nuclei, respectively, either before chemotherapy as after vincristine sulfate exposure. Future studies are needed to demonstrate the function of these two markers in conferring multidrug resistance (MDR) or predict the response to chemotherapy in canine TVT.

Immunohistochemical study of genital and extragenital forms of canine transmissible venereal tumor in Brazil

Aiming to provide insight and discussing the problems related to the diagnosis and differential diagnosis of canine transmissible venereal tumor (CTVT), especially in its extragenital form, immunohistochemical evaluation was performed and a comparison was established by analysis of the microscopic appearance of 10 genital CTVTs and 13 exclusively extragenital CTVTs previously diagnosed by cytology and histopathology. CTVTs samples were incubated with biotinylated antibodies raised against specific membrane (anti-macrophage) and cytoplasmic antigens (anti-lysozyme, anti-S-100 protein, anti-vimentin and anti-CD18) and subsequently developed using streptavidin-biotin peroxidase and streptavidin-biotin-alkaline phosphatase methods. A strong reactivity with the anti-vimentin antibody was found in 100% of the tumors tested (22/22). No reactivity was found for the anti-lysozyme, anti-macrophage, anti-S-100 protein and anti-CD18. No histopathological or immunoreactivity differences between genital and extragenital CTVTs were found. These findings do not corroborate the hypothesis of histiocytic origin of CTVT (no reactivity to anti-lysozyme, anti-macrophage and anti-CD 18 antibodies). In addition, the antibody panel used is useful to narrow the differential diagnosis for lymphomas, histiocytic tumors, amelanotic melanomas, and poorly differentiated epithelial neoplasias, among others.

Epidemiological, clinical and immunohistochemical aspects of canine lymphoma in the region of Porto Alegre, Brazil

This paper describes the epidemiological, clinical and immunohistochemical characteristics of canine lymphomas diagnosed in the region of Porto Alegre, Brazil. Thirty dogs were enrolled in the study; most of them were male (60%), mixed-breed (23%) and middle-aged or older. The majority (87%) of affected dogs showed the multicentric form. The B-cell phenotype was most frequently detected (62%); 37% of the animals were in clinical stage IV, and 83% were classified as sub-stage "b". Lymphadenopathy was observed in 67% of the cases, and dyspnea, prostration, decreased appetite and vomiting were the most common clinical signs encountered. Anemia was a frequently encountered laboratory alteration (57%), as were leukocytosis (40%), thrombocytopenia (33%), lymphopenia (30%), hyperglobulinemia (20%) and hypercalcemia (13%). The results of this study indicate that the clinical features of dogs with lymphoma in the region of Porto Alegre are similar to those observed worldwide.

Clinical and parasitological evaluation of pour-on fluazuron and ivermectin for treating canine demodicosis

The objective of the study was to evaluate the efficacy of pour-on formulations of fluazuron and ivermectin in different therapeutic protocols for treatment of demodicosis by means of quantifying mites with skin scraping, histological and clinical evaluation in dogs. Eighteen dogs with skin scrapings positive for Demodex canis were evaluated, divided into three groups. All the animals were treated every 14 days, completing 6 treatments for each animal (days 0, 14, 28, 42, 56 and 70). In group 1, pour-on 2.5% fluazuron was used at the dose of 20mg/kg; in the group 2 pour-on 2.5% fluazuron at a dose of 20 mg/kg in association with pour-on 0.5% ivermectin at the dose of 0.6mg/kg; and in group 3, pour-on 0.5% ivermectin alone was used, at the dose of 0.6mg/kg. The treatment was evaluated and monitored through skin scrapings and clinical follow-up of the lesions every 14 days for 84 days, and through histopathological examination at the end of each treatment protocol. The success rate was defined as the percentage of dogs in each group that had negative skin scrapings after the treatment: this was 16.67% for group 1, and 50% for groups 2 and 3. The reduction in mite counts reached effectiveness of 67.66%, 88.99% and 84.29% for groups 1, 2 and 3 respectively. The Wilcoxon test showed that there was a significant difference between the number of mites before and after treatment in groups 2 and 3. The histopathological examination revealed that only group 1 showed no significant difference in the intensity of infestation between days 0 and 84. Clinically, there was no significant difference between the evaluation before and after treatment in the three groups. pour-on 2.5% fluazuron and pour-on 0.5% ivermectin were not effective for treating canine demodicosis, either in association or as single therapy, when applied every 14 days for a period of 70 days. Quantification of mites using skin scrapings and histological evaluation proved to be ineffective, either one as sole therapeutic evaluation parameters, for canine demodicosis.

Identification of canine papillomavirus type 1 (CPV1) DNA in dogs with cutaneous papillomatosis

Canine oral papillomavirus (COPV), also known as Canine Papillomavirus type 1 (CPV1), induces papillomas at the mucous membranes of the oral cavity and at the haired skin of dogs. The classification of Papillomavirus (PV) types is based on the L1 capsid protein and nucleotide sequence; so far, 14 CPV types have been described in several countries, but the molecular characterization of CPV in Brazil is lacking. This study investigated the presence of the PV in seven papillomas from four mixed breed dogs from Londrina/PR, Southern Brazil, by partial sequencing of the L1 gene. Seven exophytic cutaneous lesions were surgically removed and processed for histopathological and molecular characterization. Histopathology confirmed the lesions as viral papillomas due to typical histological features. Polymerase Chain Reaction (PCR) assay using the FAP59 and FAP64 primers targeted the L1 gene followed by sequence analysis of the amplicons identified CPV1 in all evaluated papilloma samples. This study represents the first description of CPV1 DNA associated with canine papillomatosis in Brazil.

Systemic acanthamoebiasis associated with canine distemper in dogs in the semiarid region of Paraíba, Brazil

Infections by free-living amoebae can cause systemic disease in animals and humans. We describe the epidemiological, clinical and pathological aspects of disseminated acanthamoebiasis associated with canine distemper in three dogs of the semiarid region of Paraíba, Northeastern Brazil. Affected dogs developed progressive neurological and respiratory signs that progressed to death within in two to 20 days. Gross lesions were irregular and with yellow-reddish nodules randomly distributed in the lungs, heart, kidneys, spleen, lymph nodes, adrenals, and intestine. One dog had foci of malacia in the parietal cortex and another one in nucleus of brain basis. Histologically, pyogranulomas with areas of necrosis and hemorrhage in all organs affected were observed, associated with myriads of intralesional amoebic trophozoites. All three cases were concomitant canine distemper, that possibly triggered immunosuppression in the dogs. The diagnosis was performed through microscopic findings of infection by free-living amoebae and confirmed Acanthamoeba sp. by immunohistochemistry

Doenças da cavidade nasal em pequenos ruminantes no Distrito Federal e no Estado de Goiás

Resumo As enfermidades que acometem a cavidade nasal de pequenos ruminantes podem causar prejuízos aos rebanhos de ovinos e caprinos na região central do Brasil. Foi realizado estudo retrospectivo dos laudos de necropsia do Laboratório de Patologia Veterinária da Universidade de Brasília (LPV-UnB) nos anos de 2003 a 2014 para verificar a ocorrência das doenças que acometeram a cavidade nasal de pequenos ruminantes. Foram analisados 463 protocolos de ovinos e 75 de caprinos totalizando 538 casos. Seis ovinos (6/463 1,29%) foram necropsiados com rinite granulomatosa micótica ou oomicótica e 22 animais do estudo (22/538; 4,08%) tiveram o diagnóstico de oestrose, sendo 86,36% ovinos e 13,64% caprinos. As rinites piogranulomatosas em ovinos ocorreram em áreas alagadas, com abundante material vegetal em decomposição. Os ovinos com pitiose rinofacial apresentaram como principais alterações aumento de volume na região nasal devido a extensas lesões granulomatosas associadas a necrose tecidual, caracterizadas por inúmeros macrófagos e polimorfonucleares circundando centros necróticos contendo o agente envolto por reação de Splendore-Hoeppli. Os ovinos com conidiobolomicose exibiram extensas áreas de necrose e inflamação piogranulomatosa, associadas à presença de hifas fúngicas na nasofaringe e também na região peribulbar e exoftalmia. A maioria dos animais com oestrose não apresentou alterações clínico-patológicas significativas, apesar de serem encontradas larvas principalmente nos seios e conchas nasais, traqueia e seio paranasal. A importância dessas enfermidades ainda é pouco conhecida na região, sendo de grande relevância que as condições clínico-patológicas e epidemiológicas sejam elucidadas para o diagnóstico, o controle e a prevenção, para evitar a expansão e prejuízos para os rebanhos.