A polymerase chain reaction for the detection of Brucella canis in semen of naturally infected dogs

The objective was to evaluate a PCR assay for the detection of Brucella canis in canine semen, comparing its performance with that of bacterial isolation, serological tests and PCR assay of blood. Fifty-two male dogs were examined clinically to detect reproductive abnormalities and their serum was tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR assays were performed on blood and semen samples. The findings of the semen PCR were compared (Kappa coefficient and McNemar test) to those of blood PCR, culture of blood and semen, RSAT, and 2ME-RSAT. Nucleic acid extracts from semen collected from dogs not infected with B. canis were spiked with decreasing amounts of B. canis RM6/66 DNA and the resulting samples subjected to PCR. In addition, semen samples of non-infected dogs were spiked with decreasing amounts of B. canis CFU and the resulting suspensions were used for DNA extraction and amplification. of the 52 dogs that were examined, the following tests were positive: RSAT, 16 (30.7%); 2ME-RSAT, 5 (9.6%); blood culture, 14 (26.9%); semen culture, 11 (21.1%); blood PCR, 18 (34.6%); semen PCR, 18 (34.6%). The PCR assay detected as few as 3.8 fg of B. canis DNA experimentally diluted in 444.9 ng of canine DNA (extracted from semen samples of noninfected dogs). In addition, the PCR assay amplified B. canis genetic sequences from semen samples containing as little as 1.0 x 10(0) cfu/mL. We concluded that PCR assay of semen was a good candidate as a confirmatory test for the diagnosis of brucellosis in dogs; its diagnostic performance was similar to blood culture or blood PCR. Furthermore, the PCR assay of semen was more sensitive than the 2ME-RSAT or semen culture. Examination of semen by PCR should be included for diagnosis of brucellosis prior to natural mating or AI; in that regard, some dogs that were negative on serological and microbiological examinations as well as blood PCR were positive on PCR of semen. (c) 2007 Elsevier B.V. All rights reserved.

Active surveillance of canine visceral leishmaniasis and american trypanossomiasis in rural dogs from non endemic area

A leishmaniose visceral canina (LVC) e a tripanossomíase americana são importantes zoonoses para a saúde pública que encontram no cão o principal reservatório doméstico para o homem. O trabalho procurou estimar a prevalência de anticorpos circulantes anti-Trypanosoma cruzi e anti-Leishmania spp., em amostras de cães provenientes da zona rural do município de Botucatu, SP. Durante a campanha de vacinação antirrábica canina da zona rural do município, foram coletadas 689 amostras de soro e processadas pela técnica de imunofluorescência indireta. Os testes sorológicos revelaram a ausência de anticorpos anti-Leishmania spp. e, na pesquisa dos anticorpos anti-T. cruzi, foram detectados 3 (0,4%) cães.

Canine distemper virus and Toxoplasma gondii co-infection in dogs with neurological signs

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detection and molecular analysis of Toxoplasma gondii and Neospora caninum from dogs with neurological disorders

Introduction: Toxoplasma gondii and Neospora caninum are related Apicomplexa parasites responsible for systemic diseases in many species of animals, including dogs. Methods: This study aimed to determine the occurrence of T. gondii and N. caninum infections in 50 dogs with neurological signs that were admitted to the Veterinary Hospital of Universidade Estadual Paulista, City of Botucatu, Brazil. All animals were screened for antibodies using an immunofluorescent antibody test for both parasites. Tissues of positive animals were bioassayed in mice (T. gondii) and gerbils (N. caninum), and DNA was analyzed using the polymerase chain reaction (PCR). Positive samples for T. gondii by PCR were typed using restriction fragment length polymorphism-PCR for 11 markers: SAG1, SAG2 (5'-3'-SAG2 and alt.SAG2), SAG3, Btub, GRA6, L358, c22-8, c29-6, PK1 and Apico, and CS3 marker for virulence analysis. Results: Specific antibodies were detected in 11/50 (22%; 95% confidence interval (CI95%), 12.8-35.3%) animals for T. gondii and 7/50 (14%; CI95%, 7.02-26.3%) for N. caninum. In the bioassay and PCR, 7/11 (63.6%; CI95%, 34.9-84.8%) samples were positive for T. gondii and 3/7 (42.9%; CI95% I, 15.7-75.5%) samples were positive for N. caninum. Three different genotypes were identified, but only 1 was unique. Conclusions: These data confirm the presence of T. gondii and N. caninum in dogs from Brazil, indicating the importance of this host as a sentinel of T. gondii for human beings, and the genotypic variation of this parasite in Brazil.

Seroprevalence and risk factors associated with neosporosis in sheep and dogs from farms

This study aimed to establish the seroprevalence and risk factors associated with neosporosis in sheep and dogs from rural properties. 1497 blood samples were collected from sheep and 42 from dogs that cohabited with sheep from 16 farms located in the central region of São Paulo State, Brazil. For the detection of N. caninum antibodies it was performed the indirect fluorescent antibody test (IFAT >= 25). For the epidemiological study it was applied a questionnaire for the owners or responsible from the sheep and dogs regarding informations related to neosporosis. The seroprevalence obtained out of the 1497 sheep sera tested was 8.0% (CI95% = 6.7-9.2%) and out of the 42 dogs 4.8% (CI95% = 0-7.2%). Variables statistically related to seropositivity for N. caninum in sheep were: dams well as water supply (P = 0.0004; OR = 2.15), presence of other domestic canids (P = 0.0013; OR = 2.38) and presence of reproductive problems (P = 0.0031; OR = 1.75). (C) 2011 Elsevier B.V. All rights reserved.

Seroprevalence of anti-Leptospira spp. antibodies in dogs in Bahia, Brazil

The aim of this study was to determine the prevalence of anti-Leptospira spp. antibodies in dogs living in the urban area of the city of Ilheus, Bahia, Brazil using the microscopic agglutination test (MAT) to investigate 24 serovars. A semi-structured questionnaire was administered to dog owners to collect data about demography, husbandry and environmental factors. The prevalence of anti-Leptospira spp. antibodies in the population of 282 dogs was 7.1% (95% confidence interval: 4.4-10.7%). Serovar Copenhageni was the most prevalent, followed by serovars Bratislava, Canicola and Gryppotyphosa. No risk factor was detected with regard to demography (age, gender and breed), husbandry (Leptospira vaccinations, food and water exposure through their environment, hunting habits, contact with other animals and contact with rats) and environmental factors (sewage network, garbage collection, history of flooding, river proximity and wastelands). Despite the low prevalence found in this study, the seroprevalence of Leptospira spp. in healthy dogs in Ilheus indicates the presence of this agent in the environment, which may be a source of human infection. Knowledge of the serovars present in this environment is important for understanding the epidemiology of leptospirosis and establishing public health policies aimed at its control. (C) 2012 Elsevier B.V. All rights reserved.

Leishmania spp. and/or Trypanosoma cruzi diagnosis in dogs from endemic and nonendemic areas for canine visceral leishmaniasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spatial Distribution of Seropositive Dogs to Leptospira spp. and Evaluation of Leptospirosis Risk Factors Using a Decision Tree

Background: Leptospirosis is an important zoonotic disease associated with poor areas of urban settings of developing countries and early diagnosis and prompt treatment may prevent disease. Although rodents are reportedly considered the main reservoirs of leptospirosis, dogs may develop the disease, may become asymptomatic carriers and may be used as sentinels for disease epidemiology. The use of Geographical Information Systems (GIS) combined with spatial analysis techniques allows the mapping of the disease and the identification and assessment of health risk factors. Besides the use of GIS and spatial analysis, the technique of data mining, decision tree, can provide a great potential to find a pattern in the behavior of the variables that determine the occurrence of leptospirosis. The objective of the present study was to apply Geographical Information Systems and data prospection (decision tree) to evaluate the risk factors for canine leptospirosis in an area of Curitiba, PR.Materials, Methods & Results: The present study was performed on the Vila Pantanal, a urban poor community in the city of Curitiba. A total of 287 dog blood samples were randomly obtained house-by-house in a two-day sampling on January 2010. In addition, a questionnaire was applied to owners at the time of sampling. Geographical coordinates related to each household of tested dog were obtained using a Global Positioning System (GPS) for mapping the spatial distribution of reagent and non-reagent dogs to leptospirosis. For the decision tree, risk factors included results of microagglutination test (MAT) from the serum of dogs, previous disease on the household, contact with rats or other dogs, dog breed, outdoors access, feeding, trash around house or backyard, open sewer proximity and flooding. A total of 189 samples (about 2/3 of overall samples) were randomly selected for the training file and consequent decision rules. The remained 98 samples were used for the testing file. The seroprevalence showed a pattern of spatial distribution that involved all the Pantanal area, without agglomeration of reagent animals. In relation to data mining, from 189 samples used in decision tree, a total of 165 (87.3%) animal samples were correctly classified, generating a Kappa index of 0.413. A total of 154 out of 159 (96.8%) samples were considered non-reagent and were correctly classified and only 5/159 (3.2%) were wrongly identified. on the other hand, only 11 (36.7%) reagent samples were correctly classified, with 19 (63.3%) samples failing diagnosis.Discussion: The spatial distribution that involved all the Pantanal area showed that all the animals in the area are at risk of contamination by Leptospira spp. Although most samples had been classified correctly by the decision tree, a degree of difficulty of separability related to seropositive animals was observed, with only 36.7% of the samples classified correctly. This can occur due to the fact of seronegative animals number is superior to the number of seropositive ones, taking the differences in the pattern of variable behavior. The data mining helped to evaluate the most important risk factors for leptospirosis in an urban poor community of Curitiba. The variables selected by decision tree reflected the important factors about the existence of the disease (default of sewer, presence of rats and rubbish and dogs with free access to street). The analyses showed the multifactorial character of the epidemiology of canine leptospirosis.

Reduction of humeroulnar congenital elbow luxation in 8 dogs by using the transarticular pin

The purpose of this study was to evaluate the clinical and radiographic outcome in 8 dogs of surgical reduction of congenital humeroulnar luxation by using the transarticular pin. Five cases were bilateral and 3 were unilateral, for a total of 13 elbows. The treatment was performed in animals between 45 and 150 days of age. Articular stabilization was achieved by using a transarticular pin driven from the caudal aspect of the olecranon into the body of the humerus or into the distal condyle and distal metaphysis of the humerus. The follow-up period was between 1 and 19 months. There were 5 postsurgical reluxations, 3 related to the insertion of the pin into the humeral condyle and 3 related to the insertion into the humeral body. These animals needed further surgery. Six animals showed near normal return to limb function and 2 had lameness. We conclude that the use of the transarticular pin is an effective and simple method for the treatment of humeroulnar congenital elbow luxation.

The effect of season on serum testosterone concentrations in dogs

Photoperiod and environmental temperature are two important factors that may influence the reproductive cycle of various species. The objective of this study was to investigate seasonal influences on serum testosterone concentrations in dogs in a tropical zone, where the variation in day length between winter and summer solstice was approximately 2.5 h. Blood samples were collected every 15 days from seven adult dogs over a 14-month interval and serum testosterone concentrations were determined by radioimmunoassay. The year was divided into four seasons and mean testosterone concentrations for each season were related to the mean environmental temperature and rainfall during that season. Mean testosterone concentrations were 1.81 ng/mL (winter 2002), 1.93 ng/mL (spring 2002),1.31 ng/mL (summer 2003), 2.02 ng/mL (autumn 2003) and 1.93 ng/mL (winter 2003). The temperature ranged from 10.2 to 32.8 degrees C and the rainfall from 33 to 476 mm. Serum testosterone concentrations were lower in summer 2003 than in both spring 2002 (P = 0.05) and autumn 2003 (P = 0.0 16). In a tropical zone, a combination of high temperature and substantial rainfall may have reduced serum testosterone concentrations in dogs. (c) 2006 Elsevier B.V. All rights reserved.

Evaluation testicular fine needle aspiration cytology and serum testosterone levels in dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Virulence factors in Escherichia coli strains isolated from urinary tract infection and pyometra cases and from feces of healthy dogs

The aim of this study was to compare the prevalence of virulence genes in 158 Escherichia coli strains isolated from 51 clinical cases of UTIs, 52 of pyometra and from 55 fecal samples from healthy dogs by PCR. papC was found in 12 (23.5%) strains isolated from UTIs, 19 (36.5%) from pyometra and 10 (18.2%) from feces. papGII was observed in 3 (5.8%) strains from pyometra, and papGIII in 10 (19.6%) from UTIs, 15 (28.8%) from pyometra and 9 (16.4%) from feces. sfaS was detected in 22 (43.1%) strains from UTIs, 24 (46.1%) from pyometra and 19 (34.5%) from feces. hlyA was observed in 17 (33.3%) strains from UTIs, 18 (34.6%) from pyometra and 7 (12.7%) from feces, while cnf-1 was detected in 11 (21.6%) from UTIs, 21 (40.4%) from pyometra and 9 (16.4%) from feces. iucD was observed in 12 (23.5%) strains from UTIs, 9 (17.3%) from pyometra and 1 (1.8%) from feces. usp was found 17 (33.3%) isolates from UTIs and 36 (69.9%) from pyometra. (c) 2008 Elsevier Ltd. All rights reserved.

Use of extensible internal device in the femur of young dogs

An extensible internal device (EID) was developed to preserve growth plate during the treatment of fracture complications or segmental bone loss from tumour resection in children. Since this type of extensible, trans-physeal, internal fixation device has only been used in a few paediatric cases; the aim of this study was to evaluate an in vivo canine study, a surgical application of this device, and its interference with longitudinal growth of the non-fractured distal femur. Ton clinically healthy two- to three-month-old poodles weighing 1.5-2.3 kg were used. Following a medial approach to the right distal femur, one extremity of the EID, similar to a T-plate, was fixed in the femoral condyle with two cortical screws placed below the growth plate. The other extremity, consisting of an adaptable brim with two screw holes and a plate guide, was fixed in the third distal of the femoral diaphysis with two cortical screws. The EID was removed 180 days after application. All of the dogs demonstrated full weight-bearing after surgery. The values of thigh and stifle circumferences, and stifle joint motion range did not show any difference between operated and control hindlimbs. The plate slid in the device according to longitudinal bone growth, in all but one dog. In this dog, a 10.5% shortening of the femoral shaft was observed due to a lack of EID sliding. The other dogs had the some longitudinal lengths in both femurs. The EID permits longitudinal bone growth without blocking the distal femur growth plate if appropriately placed.

Osteopontin in Seminal Plasma and Sperm Membrane of Dogs

The objective of this work was to describe the presence of osteopontin (OPN) in canine seminal plasma and sperm membranes. A pool of seminal plasma and sperm membrane extract from 30 dogs was used. Polyacrylamide electrophoresis gels were performed and the bands were transferred to nitrocellulose paper and Western blot was undertaken using an antibody anti-OPN. Two and 12 bands were marked in the seminal plasma (77.2 and 15.6 kDa) and sperm membrane extracts (70.6-26.6 kDa), respectively. However, from 12 marked bands in the sperm membrane extract, only three (46.4, 37.7 and 36.5 kDa) were strongly marked. We conclude that, seminal plasma and sperm membranes from dogs contain different isoforms of OPN; yet, further studies will be necessary to determine their function in this species.