Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

The intra‐ and intertester repeatability of radiographic elbow incongruity grading is high in chondrodystrophic dog breeds

Elbow incongruity is a form of elbow dysplasia that causes osteoarthritis, pain, and lameness, and it is common in chondrodystrophic dog breeds. The objective of this retrospective secondary analysis study was to evaluate the intra‐ and interobserver repeatability of a novel radiographic incongruity grading system for assessing elbow incongruity in three chondrodystrophic dog breeds—the dachshund, Skye Terrier, and Glen of Imaal Terrier. We conducted an observer agreement study that included 220 mediolateral antebrachial radiographs from 110 dogs with the elbow in 90° flexion. The radiographs were independently assessed by three observers at three time points, using a four‐stepped grading scale. The proportion of agreement and Kappa coefficient were calculated. Both the intra‐ and interobserver proportions of agreement were substantial when three grades were required to be identical (.705‐.777 and .609, respectively), and almost perfect for two identical grades (.991‐1.000 and .991, respectively). Some differences in repeatability between breeds were noted; specifically, the intraobserver repeatability was higher in the dachshund, and the interobserver repeatability was lower in the Glen of Imaal Terrier. Our study showed that the radiographic imaging protocol and incongruity grading system have high repeatability when assessing elbow incongruity in chondrodystrophic dog breeds.     

Evaluation of a live Salmonella choleraesuis vaccine by intranasal challenge.

Weaned pigs were immunised with a live attenuated Salmonella choleraesuis vaccine. The protective immunity induced was compared with that of unvaccinated pigs by intranasal challenge with a field strain of S choleraesuis. Of 18 unvaccinated pigs, three died as a result of challenge from S choleraesuis and 10 of the survivors developed chronic lung lesions. None of the vaccinated pigs died as a result of challenge and two only showed macroscopic evidence of salmonellosis on autopsy three weeks after challenge. It was concluded that subcutaneous vaccination of pigs prevented death and clinical illness but did not prevent invasion of tissues following intranasal challenge with S choleraesuis.     

Seroprevalence and risk factors associated with Chlamydophila spp. infection in ewes in the northeast of Algeria

A cross-sectional study was carried out to estimate prevalence of Chlamydophila spp. antibodies and to investigate risk factors associated with chlamydial infection in 552 ewes between March 2011 and January 2012 in the province of Constantine. Anti-Chlamydophila antibodies were detected using an indirect enzyme-linked immunosorbent assay kit in 24.5 % of examined sera. Of the herds, 70.4 % had at least one seropositive animal. A pretested structured questionnaire was administered in order to collect information on individual animal health and herd management practices. Chi-square analysis and multivariable logistic regression model were used to identify risk factors related to Chlamydophila seropositivity. Univariable analysis revealed 17 variables with p?<?0.25 that were offered to the multivariable logistic regression model which in turn identified 12–23 months age group (OR?=?5.903, 95 % CI (OR)?=?1.690; 20.618) and not using disinfectants (OR?=?2.099, 95 % CI (OR)?=?1.314; 8.065) as risk factors for Chlamydophila spp. seropositivity. Moreover, occurrence of stillbirth problem (OR?=?3.682, 95 % CI (OR)?=?1.825; 7.430) and 5–10 % mortality rate in young lambs (OR?=?2.584, 95 % CI (OR)?=?1.058; 6.310) were significantly associated with seropositivity to Chlamydophila spp. On the other hand, availability of veterinary service was identified as a protective factor (OR?=?0.161, 95 % CI (OR)?=?0.051; 0.511).     

Effect of FMD vaccination on semen quality parameters in Karan Fries and Murrah buffalo bulls

Effect of Foot and Mouth disease (FMD) vaccination was studied on semen quality parameters of 19 Karan Fries (KF) and eight Murrah (MU) breeding bulls during the period 2002 to 2004 at Artificial Breeding Complex, NDRI, Karnal. A total of non-vaccinated 155 KF and 72 MU bulls' ejaculates were taken as control, while 169 KF and 51 MU bulls' ejaculates, collected after vaccination, were used to study the effect of vaccination stress. The results showed that FMD vaccination had no significant (P > 0.05) effect on ejaculate volume and total volume per day of semen in both KF and MU bulls. Volume of semen increased slightly during post-vaccination period in both the breeds. After FMD vaccination, there was significant (P < 0.01) decrease in mass activity (2.27 ± 0.06 vs. 1.67 ± 0.07 and 2.49 ± 0.09. vs. 1.75 ± 0.10, for KF and MU, respectively), initial motility (56.89 ± 0.03% vs. 44.62 ± 0.02% and 62.26 ± 0.04% vs. 47.08 ± 0.05%, for KF and MU, respectively), sperm concentration (754.19 ± 23.96 vs. 554.14 ± 22.95 × 10⁶/ml and 848.61 ± 33.65 vs. 571.57 ± 39.99 × 10⁶/ml, for KF and MU, respectively), and total sperm output per ejaculate (3,685.94 ± 158.40 vs. 2,781.54 ± 151.70 × 10⁶ and 2,218.75 ± 133.14 vs. 1,582.84 ± 158.20 × 10⁶, for KF and MU, respectively). Application of FMD vaccine had significantly (P < 0.05) adverse effect on most of the seminal attributes during post-vaccination in KF and MU buffalo bulls. So, the spermiograms affected following vaccination suggest that in bovines, the semen collection and preservation should be suspended till normal fertility of sperm is restored to avoid the failure of conception from artificial insemination using such semen.     

Plasma pharmacokinetics and milk levels of ceftriaxone following single intravenous administration in healthy and endometritic cows

Pharmacokinetics and milk levels of ceftriaxone were studied in healthy and endometritic cows following single intravenous administration. The drug was detected up to 8 h of dosing in plasma of healthy and endometritic cows and the drug disposition followed three-compartment open model. The values of Vd_area, AUC, t_1/2β, Cl_B, MRT and P/C ratio were 0.50 ± 0.19 L.kg⁻¹, 62.2 ± 23.3 μg.ml⁻¹.h, 1.02 ± 0.07 h, 0.30 ± 0.09 L.kg⁻¹.h⁻¹, 1.55 ± 0.25 h and 0.52 ± 0.27, respectively, in healthy and 1.55 ± 0.52 L.kg⁻¹, 37.0 ± 17.1 μg.ml⁻¹.h, 1.56 ± 0.25 h, 0.56 ± 0.14 L.kg⁻¹.h⁻¹, 2.14 ± 0.34 h and 1.44 ± 0.60, respectively, in endometritic cows. The drug was detected in milk for 36 h after administration. For MIC₉₀ of 0.5 μg.ml⁻¹ the most appropriate dosage for ceftriaxone, would be 9.0 mg.kg⁻¹ repeated at 6 h intervals for the treatment of endometritis in cows.     

Sample pooling for real-time PCR detection and virulence determination of the footrot pathogen <Emphasis Type="Italic">Dichelobacter nodosus</Emphasis>

Dichelobacter nodosus is the principal cause of ovine footrot and strain virulence is an important factor in disease severity. Therefore, detection and virulence determination of D. nodosus is important for proper diagnosis of the disease. Today this is possible by real-time PCR analysis. Analysis of large numbers of samples is costly and laborious; therefore, pooling of individual samples is common in surveillance programs. However, pooling can reduce the sensitivity of the method. The aim of this study was to develop a pooling method for real-time PCR analysis that would allow sensitive detection and simultaneous virulence determination of D. nodosus. A total of 225 sheep from 17 flocks were sampled using ESwabs within the Swedish Footrot Control Program in 2014. Samples were first analysed individually and then in pools of five by real-time PCR assays targeting the 16S rRNA and aprV2/B2 genes of D. nodosus. Each pool consisted of four negative and one positive D. nodosus samples with varying amounts of the bacterium. In the individual analysis, 61 (27.1%) samples were positive in the 16S rRNA and the aprV2/B2 PCR assays and 164 (72.9%) samples were negative. All samples positive in the aprV2/B2 PCR-assay were of aprB2 variant. The pooled analysis showed that all 41 pools were also positive for D. nodosus 16S rRNA and the aprB2 variant. The diagnostic sensitivity for pooled and individual samples was therefore similar. Our method includes concentration of the bacteria before DNA-extraction. This may account for the maintenance of diagnostic sensitivity. Diagnostic sensitivity in the real-time PCR assays of the pooled samples were comparable to the sensitivity obtained for individually analysed samples. Even sub-clinical infections were able to be detected in the pooled PCR samples which is important for control of the disease. This method may therefore be implemented in footrot control programs where it can replace analysis of individual samples.     

A spatial hazard model for cluster detection on continuous indicators of disease: application to somatic cell score

Methods for spatial cluster detection dealing with diseases quantified by continuous variables are few, whereas several diseases are better approached by continuous indicators. For example, subclinical mastitis of the dairy cow is evaluated using a continuous marker of udder inflammation, the somatic cell score (SCS). Consequently, this study proposed to analyze spatialized risk and cluster components of herd SCS through a new method based on a spatial hazard model. The dataset included annual SCS for 34 142 French dairy herds for the year 2000, and important SCS risk factors: mean parity, percentage of winter and spring calvings, and herd size. The model allowed the simultaneous estimation of the effects of known risk factors and of potential spatial clusters on SCS, and the mapping of the estimated clusters and their range. Mean parity and winter and spring calvings were significantly associated with subclinical mastitis risk. The model with the presence of 3 clusters was highly significant, and the 3 clusters were attractive, i.e. closeness to cluster center increased the occurrence of high SCS. The three localizations were the following: close to the city of Troyes in the northeast of France; around the city of Limoges in the center-west; and in the southwest close to the city of Tarbes. The semi-parametric method based on spatial hazard modeling applies to continuous variables, and takes account of both risk factors and potential heterogeneity of the background population. This tool allows a quantitative detection but assumes a spatially specified form for clusters.