Equine adipose-tissue derived mesenchymal stem cells and platelet concentrates: their association in vitro and in vivo

Equine mesenchymal stem cells (MSC) are of particular interest both for basic research and for the therapeutic approach to musculoskeletal diseases in the horse. Their multilineage differentiation potential gives them the capability to contribute to the repair of tendon, ligament and bone damage. MSCs are also considered a promising therapeutic aid in allogeneic cell transplantation, since they show low immunogenicity and immunomodulating functions.Adipose tissue-derived adult equine stem cells (AdMSC) can be isolated, expanded in vitro and then inoculated into the damaged tissue, eventually in the presence of a biological scaffold. Here we report our preliminary experience with adipose-derived mesenchymal stem cells in allogeneic cell-therapy of tendonitis in the horse. MSCs, derived from visceral adipose tissue, were grown in the presence of autologous platelet lysate and characterized for their differentiation and growth potential. Expanded AdMSC were inoculated into the damaged tendon after their dispersion in activated platelet-rich plasma (PRP), a biological scaffold that plays an important role in maintaining cells in defect sites and contributes to tissue healing. Fourteen out of sixteen treated horses showed a functional recovery and were able to return to their normal activity.     

Bovine Oocyte Quality in Relation to Ultrastructural Characteristics of Zona Pellucida,Polyspermic Penetration and Developmental Competence

In the present study, the effect of bovine oocyte quality related to ultrastructural characteristics of zona pellucida (ZP), polyspermic penetration and embryo developmental competence was evaluated. Cumulus–oocyte complexes were punctioned from 453 ovaries, classified as 1, 2, 3 and 4 according to their morphological aspect, matured for 24 h and then divided into two groups. In group A, oocytes were fixed in 2.5% glutaraldehyde and 0.1 m sodium cacodylate and examined under a scanning electron microscope. Photomicrographs were taken and ZP’s pores were evaluated in squares of 6.4‐μm width. In group B, oocytes were fertilized in vitro. After 48 h, non‐cleaved oocytes were fixed for polyspermy evaluation. On days 7, 9 and 10, embryos were classified as developed (blastocysts and hatched blastocysts). Results showed that quality 1 oocytes revealed a ZP pore diameter of 0.50 ± 0.07 μm, which was smaller than the observed on oocytes of quality 2 (0.83 ± 0.10 μm), quality 3 (1.02 ± 0.22 μm) and quality 4 (1.38 ± 0.59 μm) (p ≤ 0.05). For In Vitro Fertilization (IVF), results showed that embryos originating from oocytes classed as 3 and 4 had lower cleavage rate (68.4% and 43.8%) than those belonging to class 1 and 2 (79.5% and 69.3%) (p ≤ 0.05). None oocyte classified as 3 and 4 developed to hatch blastocysts, while for oocytes belonging to quality 1 and 2, these values were, respectively, 15.2% and 12.5%. Concerning polyspermy, oocytes class 1 and 2 had lower polyspermic penetration than those belonging to class 3 and 4 (respectively 4.1%, 4.5%, 11.1% and 9.8%, for class 1, 2, 3 and 4). In conclusion, the present study demonstrated that oocytes with low qualities result in lower developmental competence and with high percentage of polyspermy after IVF, which can be the result of the ZP structure such as the number and the pore’s diameter.     

Retrospective study of 103 presumed cases of tick (Ixodes holocyclus) envenomation in the horse

Objective Review 103 cases of presumed tick envenomation in horses. Design Retrospective study. Method Variables, including date of presentation, age, breed, weight, presence of ticks, gait and respiration scores, duration of recumbency, treatment, outcome and complications were recorded. A series of univariable screening tests were performed and used in a multivariable logistic regression model. Results There were a total of 103 cases affecting 10 breeds, aged between 1 week and 18 years of age. Horses >6 months old and weighing >100 kg had a higher odds of death than those <6 months old and <100 kg. Cases were seen from North Queensland to the central coast of New South Wales and were more likely to present in the warmer months. There was no association between the number of ticks found on an animal and death. Horses with a higher respiratory score had higher odds of dying, but there was no association between gait score and survival. Horses recumbent >120 h after presentation had higher odds of dying. Complications were reported in 35% of horses. The odds ratio for survival was higher for horses receiving >0.5 mL/kg of tick antiserum. Overall, 74% of horses survived. Multivariable modelling was limited by the small sample size. Conclusion In general, tick envenomation in horses follows the geographic distribution of Ixodes holocyclus. Tick antiserum administered at >0.5 mL/kg increases the odds of survival. It would appear that the complications associated with managing a recumbent horse increase the odds of death.     

Phage display identifies an Eastern equine encephalitis virus glycoprotein E2-specific B cell epitope

The present study identified a linear B-cell epitope in the Eastern equine encephalitis virus (EEEV) E2 glycoprotein by screening a phage-displayed random 12-mer peptide library using an EEEV E2 specific monoclonal antibody (mAb) 7C11 and defined L/F-E/R-Y-T-W-G/R-N-H/W-P as the consensus binding motif. A sequence ((321)EGLEYTWGNHPP(332)) encompassing this consensus motif was found in the EEEV E2 glycoprotein and synthesized for further epitope confirmation. Meanwhile, the corresponding epitope peptides in E2 protein of associated alphaviruses were synthesized for specificity identification. Results showed the mAb 7C11 and murine antisera all reacted strongly against the synthesized polypeptide of EEEV antigen complex, but no reaction with Western equine encephalitis virus (WEEV) and Venezuelan equine encephalitis virus (VEEV) was detected. The knowledge and reagents generated in this study may have potential applications in differential diagnosis and the development of epitope-based marker vaccines against EEEV.     

Frequency of equine laminitis: a systematic review with quality appraisal of published evidence

Equine laminitis is a highly debilitating disease of the foot. Despite its perceived importance, epidemiological characteristics are poorly understood and the true frequency of the disease remains unclear. The objective of this study was to retrospectively assess previous research to identify publications which provide the best evidence of the frequency of naturally-occurring equine laminitis. A systematic review of English language publications was conducted using MEDLINE (1950–2010), CAB Direct (1910–2010) and IVIS (1997–2010). Additional publications were included by searching bibliographies. Search terms included laminitis, equine, frequency, prevalence and incidence. Studies that allowed frequency estimations to be made for naturally-occurring equine laminitis were included. Information was extracted using predefined data fields, including 13 study quality indicators.Sixty-nine publications were appraised. Ten were considered to provide the most reliable information, estimating the frequency of equine laminitis ranging from 1.5% to 34%. Previous publications estimating laminitis frequency were generally poor quality. Laminitis frequency varied across publications however the publications included in this review focussed on many of the different underlying laminitis aetiologies and comparison of the frequencies between groups would be inappropriate. High-quality evidence-based studies are needed to estimate the true disease frequency in different equine populations.     

Efficiency of utilisations of food energy by female growing minks

The efficiency of utilisation of food energy by female growing minks, from weaning to adult age, was studied. The food given, pelleted according to an original technology, has the following chemical composition on a DM basis: 87.0% organic matter, 37.1% crude protein, 11.7% crude fat, 2.6% crude fiber, 35.6% nitrogen-free extractives and 13.0 per cent ash. Young minks had a feed intake, in relation to body weight, warying from 11.6 g to 58.6 g DM/d. Maximum feed intake related to kg0.75 was recorded at 700 g body weight (approximately 98 g DM/kg0.65). Digestibility of the given food expressed in DE, averaged 87.7 +/- 1.2%, while metabolizability, 82.3 +/- 1.1%. Total heat production related to the intaked gross energy, was 48.0 +/- 3.0%, and the retained energy, 34.3 +/- 4.0 per cent. The net efficiency of the metabolizable energy used for maintenance and production could not be accurately determined. However, taking to account the calculated values required for maintenance, of 649 kJ/kg0.75 in 300 to 600 g young minks, and of 607 kJ/kg0.75 in 600 to 1100 g young minks and also the maintenance efficiency, Km = 0.75, the coefficient for ME utilisation in protein and fat synthesis, of 0.50 and 0.75, respectively, it was able to determine the average ME efficiency used as net energy for maintenance and production: 70%. The highest values of nictemeral metabolism were recorded in the evening, and the lowest ones, at noon; the difference between the maximal and the minimal value did not exceed 6 per cent.     

Serological comparison of type strains of porcine, bovine, and ovine mycoplasmas with atypical colony morphology

The type strains of Mycoplasma hyopneumoniae, M. flocculare, M. dispar, and M. ovipneumoniae, all characterized by nipple-less colonies on solid media, were compared serologically. By indirect hemagglutination and by complement fixation tests they were found to constitute a related group. By crossed immunoelectrophoresis a sharing of common antigens was demonstrated, whereas no cross reactivity was noted by the metabolism inhibition test.The type strains of Mycoplasma hyorhinis and Mycoplasma bovirhinis were included in the study for comparison. Although some cross reaction was noted, they appeared just moderately related to the nipple-less group as well as to each other.     

Infertility in a ram associated with a knobbed acrosome abnormality of the spermatozoa

A yearling Rambouillet ram with an asymmetrical scrotum was examined for potential breeding soundness prior to use in a synchronized mating program in a purebred flock of 20 ewes. Initial sperm cell evaluation revealed 78% knobbed acrosomes associated with few other abnormalities of the head and midpiece. Use of the ram resulted in no conception in one group of ten synchronized ewes. One month later, the proportion of sperm cells with knobbed acrosomes was 80%.     

Distribution of immunoglobulin-bearing cells in the gut-associated lymphoid tissues of the turkey: effect of antibiotics

Distribution of immunoglobulin (Ig)-bearing cells in the gut-associated lymphoid tissues of antibiotic treated and untreated control turkeys (Meleagris gallopavo) was compared. Antibiotic treatment was similar to a regimen used in commercial turkey production, which included preincubation dipping of fertile eggs in gentamicin solution, injection of turkeys with gentamicin at hatching, and inclusion of chlortetracycline in the diet. Tissues were examined from turkeys at 3, 7, 14, and 21 days of age with a direct immunofluorescence procedure. Cell distribution in control turkeys was as follows: In the bursa of Fabricius, IgA-carrying cells predominated at 3 days of age, but at later intervals, the 3 classes of Ig-bearing cells were in equal numbers. In the cecal tonsils, IgM- and IgA-bearing cells were in larger numbers at 3 days of age, whereas, the IgG-bearing cells were sparsely distributed. By 7 days of age, IgM cells became more numerous in the cecal tonsils and remained numerous until 21 days of age. At 3 days of age, IgA cells predominated in the small intestines and IgM cells predominated in the large intestine. At 7 and 14 days of age, IgM cells were more numerous in the small and large intestines, but by 21 days of age, IgA cell population equaled that of IgM. The IgG cells were generally sparse in the intestines. Antibiotic treatment often resulted in lower numbers of Ig-positive cells, especially those bearing IgM and IgA. Normal development of the bursa of Fabricius was also retarded in this group.     

Coccidia in cows and calves on pasture in north Georgia (U.S.A.)

The prevalence and abundance of coccidian oocysts were determined in a herd of beef cows and calves on fescue pastures in the Piedmont area of Georgia during 4-consecutive grazing seasons. Twelve species of Eimeria were found in the feces of the calves and 10 species were found in the feces of the cows. Eimeria bovis was the most prevalent species found in both the calves and cows. It occurred in 72.5% of 1090 samples from the calves and 10.2% of 719 samples from cows. Eimeria bovis also comprised the majority of oocysts present in the fecal samples from the calves and cows. The greatest number of E. bovis in a sample was 45 800 oocysts per gram of feces (OPG) from a calf and 1900 OPG from a cow. No cases of clinical coccidiosis were seen in any of the animals sampled during the survey.