Immunoproteomic analyses of outer membrane antigens of Actinobacillus pleuropneumoniae grown under iron-restricted conditions

Actinobacillus pleuropneumoniae, a bacterial pathogen of swine and agent of porcine pneumonia, causes a highly infectious disease of economic importance in the pig industry. Commercial vaccines for A. pleuropneumoniae include whole-cell bacterins and second generation subunit vaccines but they only confer partial protective immunity. Our search for new vaccine candidates identified antigens that are expressed during conditions that mimic infection; the outer membrane (OM) proteome of A. pleuropneumoniae serotype 5b was examined under iron restriction. Quantitative profiling by 2D-DiGE technology revealed that iron restriction induced expression of previously described transferrin binding proteins (TbpA, TbpB) plus four lipoproteins including spermidine/putrescine binding periplasmic protein 1 precursor (PotD2). Immunoproteomic analyses with antisera from na?ve animals and from infected pigs were able to differentiate antigens within the OM proteome that were specifically recognized during A. pleuropneumoniae infection. Immunoblots of iron-restricted profiles detected PotD2, heme-binding protein A (HbpA), and capsule polysaccharide export protein (CpxD) as well as surface antigens TbpA, TbpB, and OmlA. These data identify OM proteins that demonstrate immunogenicity and upregulation under conditions mimicking infection, providing emphasis on lipoproteins as an important class of antigens to exploit for vaccine development for A. pleuropneumoniae.     

An evaluation of the Abaxis VSPro for the measurement of equine plasma fibrinogen concentrations

Reasons for performing study: Accurate measurement of plasma fibrinogen concentrations is an important tool for assessment of horses with inflammatory diseases. Objectives: To determine the precision and accuracy of a benchtop instrument using both fresh and frozen equine plasma by comparing the plasma fibrinogen concentration measured by a benchtop instrument to 2 separate laboratory standard methods (ACL 100 and STA Compact) for fibrinogen measurement. Methods: Accuracy and precision of the VSPro was evaluated using both human fibrinogen standards and samples from horses. Fifty frozen samples from horses with gastrointestinal disease had the fibrinogen concentration measured using the ACL 1000 and the VSPro. Fifty fresh samples were collected from hospitalised horses and fibrinogen concentration was measured using the STA Compact coagulation machine and the VSPro. Correlations for measurements were performed, as well as Bland‐Altman analysis. Results: Coefficients of variability for the VSPro ranged from 7% to 15%. The VSPro fibrinogen values were well correlated to both the ACL 1000 (r = 0.94, P<0.001) and the STA Compact measurements (r = 0.926, P<0.001). Bland‐Altman analysis showed a mean bias of ‐0.83 g/l (95% confidence interval ‐2.03–0.324 g/l) for the ACL 1000 and a mean bias of ‐0.024 g/l (95% confidence interval ‐1.434–1.386 g/l) for the STA Compact. Conclusions: The VSPro appears to have adequate accuracy and precision for clinical measurement of plasma fibrinogen concentrations. Potential relevance: The VSPro provides a measurement of equine plasma fibrinogen concentration using a benchtop instrument with a rapid test time that has comparable accuracy to the fibrinogen concentration obtained from reference laboratories.     

Heredity of navicular disease

Bone structure, nutrient canals, form and definition, and a total evaluation of both fore navicular bones were radiologically evaluated in 169 three- and four-year-old Dutch warmblood mares divided into eleven daughter (half sister) groups. On the basis of the 'total evaluation', differences between the three- and four-year-old horses were detectable. Using the 'total evaluation', significant differences were found between daughter groups consisting of three-year-old mares. These differences support the theory that podotrochleosis may be (partly) due to the presence of genetic factors. The possibilities concerning selection are presented. The results of the radiological evaluation of the stallion do not have predictive value as far as his progeny is concerned.     

Effects of calcium salts of long-chain fatty acids and rumen-protected methionine on plasma concentrations of ghrelin, glucagon-like peptide-1 (7 to 36) amide and pancreatic hormones in lactating cows

Our objective was to determine the effects of calcium salts of long-chain fatty acids (CLFAs) and rumen-protected methionine (RPM) on plasma concentrations of ghrelin, glucagon-like peptide-1 (7 to 36) amide, and pancreatic hormones in lactating cows. Four Holstein cows in midlactation were used in a 4 by 4 Latin square experiment in each 2-wk period. Cows were fed corn silage-based diets with supplements of CLFAs (1.5% added on dry matter basis), RPM (20 g/d), CLFAs plus RPM, and without supplement. Jugular blood samples were taken from 1 h before to 2 h after morning feeding at 10-min intervals on day 12 of each period. CLFAs decreased dry matter intake, but RPM did not affect dry matter intake. Both supplements of CLFAs and RPM did not affect metabolizable energy intake and milk yield and composition. Plasma concentrations of NEFAs, triglyceride (TG), and total cholesterol (T-Cho) were increased with CLFAs alone, but increases of plasma concentrations of TG and T-Cho were moderated by CLFAs plus RPM. Calcium salts of long-chain fatty acids increased plasma ghrelin concentration, and the ghrelin concentration with CLFAs plus RPM was the highest among the treatments. Plasma concentrations of glucagon-like peptide-1, glucagon, and insulin were decreased with CLFAs, whereas adding RPM moderated the decrease of plasma glucagon concentration by CLFAs. These results indicate that the addition of methionine to cows given CLFAs increases plasma concentrations of ghrelin and glucagon associated with the decrease in plasma concentrations of TG and T-Cho.     

Cover Image,Volume 16, Issue 3

The cover image, by K. Sakai et al., is based on the Original Article Association of tumour‐infiltrating regulatory T cells with adverse outcomes in dogs with malignant tumours, DOI: 10.1111/vco.12383 .

     

Between-founder heterogeneity in inbreeding depression for production in Jersey cows

Severity of inbreeding depression depends on the hidden (i.e., recessive) genetic load carried by a population. If the load is distributed unevenly among founder genomes, or founder-lines were exposed to variable amounts of selection, descendants from different founders may be differentially affected by inbreeding. Between-founder heterogeneity in inbreeding depression for production traits and somatic cell score in milk (SCS) was studied using records from 59,788 Jersey cows. Inbreeding coefficients (F) were partitioned into components due to four founders, plus a remainder. A two-stage statistical analysis was performed. First, empirical best linear unbiased predictions (EBLUP) of residuals for milk, fat and protein yield, and for SCS, were computed using linear models including fixed effects of herd–year–season, age at calving and days in milk, and random additive genetic effects of individual cows. Second, models with total and partial inbreeding coefficients as predictor variables were fitted to EBLUP residuals, for each trait. Tests of differences between slopes indicated that regressions of milk, fat and protein yield on partial inbreeding coefficients were heterogeneous; SCS did not exhibit inbreeding depression. Hence, alleles contributing to inbreeding depression for production in this Jersey population seem to be associated with specific founders. This indicates that a homogeneous effect of inbreeding on production may be an incorrect statistical specification in genetic evaluation models that attempt to account for inbreeding depression. Furthermore, the observed variability between effects of partial inbreeding due to different founders implies that inbreeding effects on yield traits may be due to alleles with major effects.