Development and characterization of mouse monoclonal antibodies reactive with chicken CD83

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein containing the extracellular region of chCD83 was expressed in Chinese Hamster Ovary (CHO) cells and isolated from the spent cell culture medium by protein G affinity chromatography. The extracellular region of the chCD83 protein was purified and used to immunize mice. A cell fusion was performed, from which 342 hybridomas were screened for mAbs to chCD83. Two mAbs, chCD83-159 and chCD83-227, stained the greatest percentage of chCD83-transfected CHO cells and were selected for further characterization. By flow cytometry, both mAbs reacted with a chicken macrophage cell line, HD11. Both mAbs also recognized a single 53 kDa protein on Western blots of lysates from lipopolysaccharide-stimulated spleen mononuclear cells or unstimulated HD11 cells. Immunostaining of chicken secondary lymphoid organs identified chCD83(+) cells with morphologic and subtissue localization properties comparable to mammalian DCs. In vitro stimulation of spleen mononuclear cells with concanavalin A (Con A) decreased the percentage of chCD83(+) cells compared with cells treated with medium alone. Interestingly, spleen cells treated with Con A in the presence of chCD83-227 mAb exhibited decreased percentage of MHCII(+) cells compared with cells treated with an isotype-matched negative control mAb. These chCD83 mAbs may be useful for future investigations of chicken immune cell maturation and mechanisms of action.     

Evaluation of bee venom and hyaluronic acid in the intra-articular treatment of osteoarthritis in an experimental rabbit model

The aim of this study was to investigate bee venom and hyaluronic acid in the intra-articular treatment of osteoarthritis in an experimental rabbit model. Forty-five rabbits were used and they were randomly divided into three groups (BVI, BVII, and HA) and each group was divided to two subgroups to evaluate the radiologic, magnetic resonance imaging, histopathologic, and biochemical evaluation in post treatment second week (a) and twelfth week (b). Radiologically, a significant difference was observed in the HA group (P<0.05). The MRI evaluation of at any time in group BVI(b) was found to be different. No significant differences were seen between the groups, biochemically. Histopathologically, cellularity, and orthochromasia was evident with Safranin-O in the BVI(b) and BVII(a); adhesions were seen in the BVII(a) group and clustering of chondrocyte in the HA(b) group were found to be different. Consequently, intra-articular application of HA and BV for experimental model of osteoarthritis has no significant influence upon recovery after therapy.     

Experimental field trial with an immunostimulating complex (ISCOM) vaccine against contagious bovine pleuropneumonia

The experimental field trial with an immunostimulating complex (ISCOM) vaccine has been an occasion to explore the role of a Th1 response in the pathogenesis caused by Mycoplasma mycoides subsp. mycoides small colony (MmmSC) and in immune protection. The ISCOM complex is known to promote Th1 response. Antibodies to MmmSC were detected by indirect enzyme-linked immunosorbent assay (ELISA) in the vaccinated cattle, although the levels were lower than in a previous study. No antibodies were detected by complement fixation test (CF). After the challenge infection, vaccinated animals developed CF antibody response. They showed significantly reduced mortality compared with controls. However, gross pathological and histopathological score for vaccinated animals was as high as for the non-vaccinated, characterized by a high inflammatory reaction with histopathology dominated by interlobular pneumonia with vasculitis.     

Diurnal variation and the effect of feed restriction on plasma and milk metabolites in TMR-fed dairy cows

The objective was to study the diurnal variation in metabolites in plasma and milk of dairy cows fed total mixed rations (TMR) with a low-energy (LE) or high-energy content (HE) expected to give a minor and a major diurnal variation, respectively. Further, the purpose was to quantify and compare the responses in plasma and milk parameters when cows changed from ad libitum to restrictive feeding. Eight multiparous, early-lactating Danish Holstein cows were used in a cross-over design with two consecutive 14-day periods. Blood and milk samples were collected hourly on day 11 of each period and on days 12-14 of each period, the cows were fed restrictively (65% of ad libitum dry-matter intake). The concentration of beta-hydroxybutyrate (BHB) in plasma was significantly higher in the evening for cows fed the HE TMR, than for cows fed the LE TMR. There was a significant diurnal variation in BHB in milk, with the highest concentrations between milkings and the lowest concentrations at milking. Non-esterified fatty acids (NEFA) in plasma showed significant diurnal variation that was caused by high concentrations in the morning. Plasma glucose did not show any diurnal variation. It has been argued that feeding a TMR removes diurnal changes related to feeding, which is contrary to earlier diurnal studies where concentrates have been fed twice daily. Feed restriction increased (P < 0.001) NEFA and BHB in plasma by 121 and 90%, respectively, while the glucose concentration decreased (P < 0.001) by 19%. Milk concentrations of BHB, citrate and fat increased (P < 0.001) by 163, 11 and 26%, respectively, because of feed restriction, while there were no changes in milk protein and lactose. The relatively high increase in BHB during feed restriction suggests that BHB is more advantageous as a milk indicator of metabolic status in dairy cows than citrate and fat.     

Multivariate analysis of sexual size dimorphism in local turkeys (Meleagris gallopavo) in Nigeria

Sexual size dimorphism is a key evolutionary feature that can lead to important biological insights. To improve methods of sexing live birds in the field, we assessed sexual size dimorphism in Nigerian local turkeys (Meleagris gallopavo) using multivariate techniques. Measurements were taken on 125 twenty-week-old birds reared under the intensive management system. The body parameters measured were body weight, body length, breast girth, thigh length, shank length, keel length, wing length and wing span. Univariate analysis revealed that toms (males) had significantly (P < 0.05) higher mean values than hens (females) in all the measured traits. Positive phenotypic correlations between body weight and body measurements ranged from 0.445 to 0.821 in toms and 0.053–0.660 in hens, respectively. Three principal components (PC1, PC2 and PC3) were extracted in toms, each accounting for 63.70%, 19.42% and 5.72% of the total variance, respectively. However, four principal components (PC1, PC2, PC3 and PC4) were extracted in hens, which explained 54.03%, 15.29%, 11.68% and 6.95%, respectively of the generalised variance. A stepwise discriminant function analysis of the eight morphological traits indicated that body weight, body length, tail length and wing span were the most discriminating variables in separating the sexes. The single discriminant function obtained was able to correctly classify 100% of the birds into their source population. The results obtained from the present study could aid future management decisions, ecological studies and conservation of local turkeys in a developing economy.     

Post mortem survey of peripheral dental caries in 510 Swedish horses

Reasons for performing study: Peripheral caries (PC) of equine teeth is a poorly described disorder that can cause serious clinical problems if it progresses. Objectives: To assess the prevalence, sites and severity of PC in a population of Swedish horses. Methods: A post mortem study of 510 equine skulls was performed in 2 Swedish equine abattoirs. Results: PC only affected the cheek teeth (CT) and was present in 6.1% (31/510) of skulls. It affected mainly the peripheral cementum, and 87% of PC in the 29 affected mature horses occurred in the 3 caudal CT (Triadan 09‐11). Concurrent infundibular caries involving most maxillary CT (mean 9.7/skull) was present in 32% of skulls affected with PC. Trotting horses (mean age 8.1 years) believed to be on a high concentrate and silage diet were preferentially affected with PC in this population. Food was usually tightly adherent to the PC lesions and this feature may have promoted the progression of the disease. Significantly increased levels of diastemata were present in PC‐affected horses, and periodontal disease was present in areas adjacent to some PC lesions. Conclusions: PC is a relatively common disorder of horses under certain management conditions that can progress to cause serious dental disorders, especially if concurrent, widespread infundibular caries is present. Potential relevance: Equine clinicians should be aware of this significant dental disorder and research into its aetiopathogenesis, possible prevention and treatment are required.     

Feline Sporotrichosis in the Southern Region of Rio Grande Do Sul,Brazil: Clinical,Zoonotic and Therapeutic Aspects

Clinical cases of feline sporotrichosis, originating in the Pelotas region and diagnosed at the Laboratory of Infectious Diseases (UFPel), were studied in the period from 2002 to 2006. The animals were evaluated according to the clinical forms of the mycosis, time of lesion appearance, severity of the clinical diagnosis and evolution of cutaneous lesions throughout the treatment period. Mycological analyses, carried out through direct examination, cultivation of tissue samples and exudates of feline lesions all confirmed the diagnosis of sporotrichosis in the 15 animals under study. The cutaneous dissemination form was observed in 10 animals, of which three showed prostration, anorexia and dehydration. The zoonosis occurred in 20% of case studies, and the pet owners and one attendant at a veterinary clinic were infected, developing the fixed and disseminated cutaneous forms. The treatment of mycosis was carried out with itraconazole, 10 mg kg^?1, once a day, on 12 animals. The cure of the clinical symptoms was observed on 50% of the felines. This study demonstrates a good clinical response of felines with sporotrichosis, when they were treated itraconazole and calls the attention for the incidence of human sporotrichosis on people related to the veterinary activity as well as for pet owners.     

Comparisons of 3‐, 2‐Dimensional,and M‐Mode Echocardiographical Methods for Estimation of Left Chamber Volumes in Dogs with and without Acquired Heart Disease

Background: Real‐time 3‐dimensional echocardiography (RT3D) is a recent technique based on volumetric scanning, eliminating the need for geometric modeling of the cardiac chambers and minimizing the errors caused by foreshortened views. Hypothesis: Estimations of left ventricular (LV) end‐diastolic (EDV) and end‐systolic volume (ESV), and left atrial (LA) size, differ depending on the echocardiographic technique of estimation. Animals: Fifty‐one dogs with acquired heart disease and 34 healthy control dogs. Methods: Prospective observational study by M‐mode (Teichholz method), Simpson's modified 2‐dimensional (2D) method, and RT3D methods for estimation of LV volumes. LA size was evaluated by 2D and RT3D methods. Results: RT3D showed good agreement with 2D for EDV and ESV, whereas Teichholz method overestimated LV volumes in comparison with the other 2 methods by approximately a factor 2. There were no statistically significant differences among the 3 methods in estimating ejection fraction. Comparison between RT3D assessment of LA end‐systolic volume per kilogram (LAs/kg) and LA to aortic ratio (LA/Ao) measured by 2D relative to each other showed that the RT3D method underestimated LAs/kg at lower values, and overestimated it at higher values. The difference between methods increased with increasing LA size. Conclusions and Clinical Importance: There was good agreement between RT3D and 2D methods of estimating EDV and ESV, whereas the Teichholz method overestimated LV volumes by approximately a factor 2. In comparison with RT3D, LA/Ao underestimated LA size, especially when LA was enlarged.     

Risk factors associated with Neospora caninum seropositivity in randomly sampled Canadian dairy cows and herds

Our objective was to determine cow- and herd-level risk factors associated with seropositivity for Neospora caninum in a large number of randomly selected Canadian dairy herds, controlling for important confounding variables and co-infections with bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV) and Mycobacterium avium subspecies paratuberculosis (MAP). Serum samples were obtained from 30 randomly selected cows, where available, in 240 herds using monthly milk testing, within 6 of 10 provinces, and these samples were tested for antibodies against BLV, MAP and N. caninum using commercially available ELISA test kits. Five unvaccinated cattle >6 months old from each herd were tested for antibodies to BVDV using virus neutralization. Most herd-level predictors were obtained through personal interviews with questionnaires administrated to each farm manager. A mixed logistic-regression model was built using N. caninum serostatus at the cow-level as the outcome variable, with herd as a random effect and province as a fixed effect. A BLV seropositive cow was 1.50 times more likely to be seropositive for N. caninum than a BLV-seronegative cow, and this was the only cow-level variable to remain in the final model. Regarding herd-level variables, with “no on-farm dogs” as the baseline, “presence of dogs but not known to eat placentas and/or fetuses” increased the odds of seropositivity for N. caninum by a factor of 1.66. For “presence of dogs known to eat placentas and/or fetuses”, the odds ratio (OR) was 2.75, demonstrating a dose–response relationship. “Using embryo transfer” (OR = 0.69), “asking for a BVDV-negative test before introducing an animal” (OR = 0.30), “using monensin in dry cows” (OR = 0.71), and “heifers having nose-to-nose contact with calves” (OR = 0.73) were all dichotomous variables negatively associated with seropositivity for N. caninum. “Number of milk cows on the farm” (OR = 0.99), and “area (acres) used for forage production” (OR = 0.99) were continuous variables negatively associated with N. caninum seropositivity.