Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological findings

Clinical signs, humoral and cellular immune responses, and microscopic and gross tissue alterations resulting from acute experimental Ehrlichia canis infection in dogs were studied. Four dogs were inoculated with E. canis and four were used as uninfected controls. After a 10-14-day incubation period, infected dogs developed pyrexia up to 41 degrees C for 6-8 days. Antibody titers to E. canis antigen were demonstrable in all inoculated dogs at 30 days post-infection. Necropsy of infected animals revealed pale mucous membranes, generalized lymphadenopathy, splenomegaly, edema and ascites. Microcopically, the main lesions were: lymphoreticular hyperplasia in cortical areas of lymph nodes and spleenic white pulp, periportal accumulation of mononuclear cells and centrolobular fatty degeneration of the liver. Kidneys presented with glomerulonephritis characterized by interstitial mononuclear infiltration. Immunophenotyping of lymphocytes from lymph nodes and spleen sections displayed alterations in IgG, IgM, CD3+ and CD8+ cells population in infected dogs.     

Flow cytometric assay in peripheral blood of dogs--reference values for leukocytes from Brazilian beagles

Use of domestic reference values in the flow cytometry analysis is known to improve its accuracy by integrating local variations as gender, race and age. Up to date application of flow cytometry in veterinary medicine has been limited to describe the percentual values just for peripheral lymphocytes subsets of blood. We now report establishment of reference values for a wide range of proportional and absolute numbers of peripheral blood leukocytes, including T cells subsets, B cells, monocytes and eosinophils, applicable to the healthy population of Beagles in Brazil and other regions with similar demographic characteristics. Normal reference values were also established to estimate the gender-related differences. This information will provide clinical aid in the evaluation of immunologic status as well as standard values for experimental animals of dogs from Brazil and other similar regions.     

Molecular characterization of canine coronavirus strains circulating in Brazil

To characterize canine coronavirus (CCoV) circulating in diarrheic puppies in Brazil, 250 fecal samples collected between 2006 and 2012 were tested. By using RT-PCR to partially amplify the M gene, CCoV RNA was detected in 30 samples. Sequence analysis of the M protein grouped eight strains with CCoV-I and another 19 with CCoV-II prototypes. To genotype/subtype the CCoV strains and assess the occurrence of single or multiple CCoV infections, RT-PCR of the S gene was performed, and 25/30 CCoV-positive strains amplified with one or two primer pairs. For 17/25 samples, single infections were detected as follows: six CCoV-I, nine CCoV-IIa and two CCoV-IIb. Eight samples were positive for more than one genotype/subtype as follows: seven CCoV-I/IIa and one CCoV-I/IIb. Sequence analysis revealed that the CCoV-I and IIa strains shared high genetic similarity to each other and to the prototypes. The Brazilian strains of CCoV-IIb displayed an aminoacid insertion that was also described in CCoV-IIb-UCD-1 and TGEV strains. Among the 25 CCoV-positive puppies, five had a fatal outcome, all but one of which were cases of mixed infection. The current study is the first reported molecular characterization of CCoV-I, IIa and IIb strains in Brazil.     

Effect of Breeding Activity on the Microflora of the External Genitalia and in the Semen of Stallions,and the Relationship Between Micro‐organisms on the Skin and on the External Genitalia

A possible role of breeding activities in the composition of the microbial population in stallions' external genitalia (EG) and the relationship between micro‐organisms colonizing the skin of the abdomen and the ones colonizing the EG have not been studied. In experiment 1, EG microbiological samples were collected from 41 stallions used for both natural cover and semen collection (BST) and from 18 non‐breeding stallions (NBST). A higher (p < 0.05) frequency of isolation of potentially pathogenic species was found for BST. Age did not influence number of micro‐organism species isolated both in BST and NBST. In experiment 2, the microbial content of the EG and semen was compared in 23 BST. Most micro‐organisms isolated from the EG were present in semen, albeit with a numerically lower prevalence. In 7 stallions, six microbial species isolated from semen were absent from the EG cultures, suggesting contamination by the operator. In experiment 3, a numerically higher number of micro‐organism species was isolated from the EG of 31 stallions, than from their skin of the ventral abdomen in contact with the penis or from the skin of the thorax. With the sole exception of Escherichia coli, potentially pathogenic bacteria were only isolated from the EG but not from the skin. Results suggest that breeding activity increased the number of species colonizing the EG; most species isolated from the EG were also found in semen even if with a lower frequency, and additional semen contamination seemed to occur during its manipulation. Many micro‐organism species of the skin were also isolated from the penis, but independently of being or not in contact with the penis, skin did not seem to provide an adequate environment for the growth of potentially pathogenic bacteria that were isolated from EG, with the sole exception for E. coli.     

Clinical value of carcinoembryonic antigen in mammary neoplasms of bitches

This study aimed at evaluating the behaviour and understanding the diagnostic value of the carcinoembryonic antigen (CEA) in bitches with mammary carcinoma as a tool for monitoring and prognosis of canine cancer patients. Serum samples from 77 bitches were divided into four groups, G1 (n = 21), control group (healthy/neoplasia free bitches); G2 (n = 31), bitches with non‐metastatic mammary carcinoma less than 3 cm; G3 (n = 12), bitches with non‐metastatic mammary carcinoma greater than 3 cm; and, G4 (n = 13) bitches with mammary carcinoma and lymph node metastasis. The marker was dosed once in G1, whereas in G2, G3 and G4, CEA levels were determined before (M0) and 15 days after (M1) mastectomy, using the ELISA kit for humans while reading used ELISYS ONE human. A group of 11 bitches was followed up 45 days after mastectomy (M2). The results for the concentration of markers in blood serum samples at the evaluated times and their relationship with neoplasia biological behaviour and observed clinicopathological changes were evaluated by the Tukey test at 5% significance. The ROC curve was established to find the cut‐off value and calculate the test sensitivity and specificity, the multivariate matching analysis was performed to confirm the association between CEA values and clinicopathological variables. CEA values increased significantly in bitches with mammary carcinoma, metastatic tumours with a diameter larger than 3.0 cm and high grade, compared with healthy ones. In addition, mastectomy reduced the CEA concentration in the blood (P < .05) whereas high CEA levels were associated with unfavourable prognostic factors (P < .05). The biomarker presented good diagnostic value, especially for more aggressive tumours. In conclusion, CEA serum concentrations allowed to follow efficiently the evolution of mammary tumours in bitches, since CEA values increased in bitches with mammary gland tumour and decreased after mastectomy while correlating with prognostic factors such as tumour size, nodal metastasis and histological grade. Further studies are still needed to confirm its diagnostic value for follow‐up of relapse and early metastasis.     

SHORT COMMUNICATION: Neutrophil oxidative metabolism and haemogram of sheep experimentally infected with Haemonchus contortus and supplemented with selenium and vitamin E

The objective of this study was to evaluate neutrophil oxidative metabolism and haemogram in sheep experimentally infected with Haemonchus contortus and supplemented with selenium and vitamin E. Twenty male Corriedale sheep were utilised and distributed into four experimental groups each with five animals infected with larvae: G1‐ supplemented with sodium selenite, 0.2 mg/kg body weight (bw) given intramuscularly (IM); G2‐ supplemented with sodium selenite and vitamin E, 0.2 mg/kg bw and 2000 IU per animal, respectively, both IM; G3‐ supplemented with vitamin E, 2000 IU per animal IM; G4‐ not supplemented. A haemogram and the number of parasite eggs were determined in samples of blood and faeces, respectively, on days zero (T0), 20 (T1), 40 (T2) and 60 (T3) and nitroblue tetrazolium (NBT) assays were performed on heparinized blood samples taken on days zero, 30 and 60. A lower total leucocyte count was detected in G1 in relation to G4 at T4. Lymphocytes were reduced in G1 in relation to G3 and G4 at T3. In both non‐stimulated (NBT‐NS) and stimulated (NBT‐S) dye reduction assays, there was reduced activity at 60 days, in relation to other times in the groups treated with selenium (G1 and G2). On the basis of results obtained, we conclude that supplementation with selenium provides better antioxidant protection to neutrophils.     

Genome-wide association study for age at puberty in young Nelore bulls

Selection for bulls that would reach puberty early reduces the generation interval and increases fertility and herd productivity. Despite its economic importance, there are few QTL associated with age at puberty described in the literature. In this study, a weighted single-step genome-wide association study was performed to detect genomic regions and putative candidate genes related to age at puberty in young Nelore bulls. Several protein-coding genes related to spermatogenesis functions were identified within the genomic regions that explain more than 0.5% of the additive genetic variance for age at puberty in Nelore bulls, such as ADAM11, BRCA1, CSNK2A, CREBBP, MEIOC, NDRG2, NECTIN3, PARP2, PARP9, PRSS21, RAD51C, RNASE4, SLX4, SPA17, TEX14, TIMP2 and TRIP13 gene. Enrichment analysis by DAVID also revealed several GO terms related to spermatogenesis such as DNA replication (GO:0006260), male meiosis I (GO:0007141), double-strand break repair (GO:0006302), base excision repair (GO:0006284), apoptotic process (GO:0006915), cell–cell adhesion (GO: 0098609) and focal adhesion (GO:0005925). The heritability for age at puberty shows that this trait can be improved based on traditional EBV selection. Adding genomic information to the system helps to elucidate genes and molecular mechanisms controlling the sexual precocity and could help to predict sexual precocity in Nelore bulls with greater accuracy at younger age, which would speed up the breeding programme for this breed.     

Rural veterinary services in Western Australia: Part B. Rural practice

OBJECTIVE: To determine the current status of rural veterinary services in Western Australia. PROCEDURE: A questionnaire was sent to all eligible rural practitioners registered in 2006 and the replies were transferred to Microsoft Excel for analysis. RESULTS: Of the rural practitioners invited to participate in the survey replies were received from 67%. There were equal numbers of females and males. Their mean age was 44 years. Ninety per cent of respondents considered knowledge gained as an undergraduate was sufficient to equip them for practice, but only 60% considered their practical skills adequate. Thirteen per cent of those in rural practices in 2005 had left by 2006. Eighty-nine per cent of respondents were in mixed practice, the balance in specific species practice, such as equine, large animal and production animal consultancy. The majority of rural practitioners relied on servicing companion animals for their viability; 7% earned their income from servicing production animals only. Seventy per cent utilised merchandising and the sale of pet foods to supplement the income received from the traditional veterinary services and 34% found it necessary to earn an independent income. A quarter considered that rural practice did not have a future. CONCLUSION: The majority of rural practitioners in Western Australia depend on companion animals, not production animals, to remain viable, with very few operating production animal services. Poor remuneration is a major reason why veterinarians leave rural practice, and many find it necessary to supplement their income or develop an independent income.