Evaluation of parentage testing in the Turkish Holstein population based on 12 microsatellite loci

In the present study, 12 microsatellite loci (ETH10, ETH225, ETH03, TGLA122, TGLA227, BM1824, BM2113, INRA23, SPS115, TGLA126, RM006 and BM1818) were evaluated for their possible use to confirm selected pedigree relationships between 7 bulls, their 21 male offspring, and their 64 second-generation female offspring within the progeny test started in Turkey. The nine loci (BM1824, INRA23, BM2113, SPS115, ETH10, TGLA122, ETH225, TGLA126 and TGLA227) recommended by ISAG displayed high values for the measures of informativeness (allele numbers, heterozygosity, polymorphic information content, frequency of the most common allele, and power of discrimination). When both parents are known calculated combined probability of exclusion was at least 0.999. Range of probability of paternity (POP) values were 0.814–0.9999. Except 3 cases (4.7%), the alleged paternity relationships were confirmed. To have a higher confidence in POP values new loci must be integrated into the set of 9 loci used.     

Molecular characterization and differentiation of five horse breeds raised in Algeria using polymorphic microsatellite markers

In this study, genetic analyses of diversity and differentiation were performed on five horse breeds raised in Algeria (Barb, Arab‐Barb, Arabian, Thoroughbred and French Trotter). All microsatellite markers were highly polymorphic in all the breeds. A total of 123 alleles from 14 microsatellite loci were detected in 201 horses. The average number of alleles per locus was the highest in the Arab‐Barb horses (7.86) and lowest in the thoroughbred breed (5.71), whereas the observed and expected heterozygosities per breed ranged from 0.71 (Thoroughbred) to 0.752 (Barb) and 0.71 (Thoroughbred) to 0.77 (Arab‐Barb), respectively. The genetic differentiation between the breeds was significant (p < 0.01) based on the infinitesimal model (F_ST). Three different approaches for evaluating the genetic relationships were applied. Genetic distances, the factorial correspondence analysis and structure analysis showed that a significant amount of genetic variation is maintained in the native horse populations and the other breeds. The Barb and Arab‐Barb breeds seem to be the most genetically related and support the decision to consider the breeds as same population.     

采用血液遗传标记对马亲仔关系案例的检测与分析

采用16个血液遗传住点对北京华骏育马场的275组及中国马术协会所提供的2组马匹进行了亲仔关系的鉴定，并对其中典型的8组马匹进行了分析。得出结果如下：6匹幼驹与其指定父母双方均不符而被排除亲仔关系，经验证确认为属于幼驹被串换；在所检的遗传住点中，以D血型、运铁蛋白(Tf)、蛋白酶抑制剂(Pi)和6-磷酸葡萄糖脱氢酶(6-PGD)检测亲仔关系错误的效果最好；在根据自身外貌无法断定为非纯血马的情况下，采用从非纯血马亲本来鉴定是行之有效的。     

Analysis of genetic diversity and the determination of relationships among western Mediterranean horse breeds using microsatellite markers

The distribution of genetic diversity and the genetic relationships among western Mediterranean horse breeds were investigated using microsatellite markers. The examined sample included seven Spanish and three Italian local horse breeds and populations, plus a Spanish Thoroughbred outgroup. The total number of animals examined was 682 (on average 62 animals per breed; range 20–122). The microsatellite marker set analysed provided 128 alleles (10.7 alleles per locus). Within‐breed genetic diversity was always high (>0.70), with breeds contributing about 8% of the total genetic variability. The mean molecular coancestry of the entire population examined was 0.205, Losino being the breed that contributed most. In addition to Nei's standard and Reynolds’ genetic distances, pair‐wise kinship distance and molecular coancestry were estimated. Remarkably similar breed rankings were obtained with all methods. Clustering analysis provided an accurate representation of the current genetic relationships among the breeds. Determining coancestry is useful for analysing genetic diversity distribution between and within breeds and provides a good framework for jointly analysing molecular markers and pedigree information. An integrated analysis was undertaken to obtain information on the population dynamics in western Mediterranean native horse populations, and to better determine conservation priorities.     

A comparison of intradermal testing and detection of allergen-specific immunoglobulin E in serum by enzyme-linked immunosorbent assay in horses affected with skin hypersensitivity

Skin hypersensitivities (allergies) in horses are often diagnosed using clinical signs only. Intradermal testing or serological assays are diagnostic options to confirm the allergic nature of the disease and to identify the allergen(s). Our objective was to develop an allergen-specific enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for horse IgE and to examine its potential for allergen detection in serum in comparison to intradermal testing. Intradermal testing with 61 allergen extracts was performed on 10 horses affected with skin hypersensitivity. Their sera were analyzed by ELISA for IgE antibodies to the same allergens. The kappa test of concordance was used for comparison of the results of both tests. Out of 61 allergen extracts, only two (Timothy and Quack) had kappa values greater than 0.60, suggesting a substantial agreement between skin testing and IgE ELISA. The statistical comparison of the remaining 59 allergens showed little or no concordance between the tests beyond chance. To identify parameters that may influence the sensitivity of the ELISA, the assay was modified to detect allergen-specific IgGb and IgG(T) in serum, and the protein content in all allergen extracts was determined by SDS-PAGE. The commercial allergen extracts revealed a high variation in detectable protein. High concentrations of allergen-specific IgG in horse serum were found to compete with IgE for binding to the plates. In conclusion, an ELISA using whole serum and crude allergen preparations provides limited diagnostic information in horses. The reliable diagnosis of allergens in equine skin hypersensitivity is essential to improve allergen-specific treatments, such as hyposensitization, or the development of allergy vaccines.     

Multiple forms of alkaline phosphatase in horse bone,liver, kidney,duodenum, caecum and serum separated by agarose gel electrophoresis with and without neuraminidase pretreatment

Horse bone, liver, duodenum, caecum and kidney alkaline phosphatases were separated by a commercial agarose gel electrophoresis method with and without neuraminidase pretreatment, following the manufacturer's directions. Tissue extracts were obtained in saline solution and ALP extracted from cell membranes by the butanol method. Electrophoresis was performed using a TRIS/barbital/sodium barbital buffer with detergent, pH 8.6 to 9.0, at 250 V for 30 minutes. Bone, liver and kidney untreated extracts showed two ALP bands each, but with different relative migration (compared to albumin migration). When they were preincubated with neuraminidase, the two bone bands showed a marked decrease in their migration, followed by the kidney ALP bands and the most anodic band of liver Both intestinal untreated extracts showed three bands but with different mobilities. After preincubation with neuraminidase, the three bands of caecum mucosa decreased in their migration, and the most anodic duodenum band disappeared, overlapping the second one. When tissue extracts were incubated with wheat germ-lectin (WGL), 74.5% of bone extract ALP and 67.2% of caecum extract ALP precipitated, which demonstrated that the ALP band of both tissues have similar groups in the carbohydrate side chains. Horse serum showed two electrophoretic bands, which increased to three bands when treated with neuraminidase. ALP from hepatocytes was the dominant isoform, followed by a caecum band. Because the electrophoretic mobilities of some of the tissue bands studied were identical, the neuraminidase agarose electrophoretic method appeared to be a satisfactory alternative to separate them.     

The Effect of Pulsed Electromagnetic Fields on Back Pain in Polo Ponies Evaluated by Pressure Algometry and Flexion Testing—A Randomized,Double-blind,Placebo-controlled Trial

This study assessed the therapeutic effect of pulsed electromagnetic fields (PEMF) on the backs of Polo ponies by measuring mechanical nociceptive thresholds (MNTs) and induced back movement. Twenty Polo ponies in regular training and competition were assigned to two groups. A double-blind, placebo-controlled, crossover field study with PEMF was performed, consisting of two 10-day therapy periods. At the beginning and end of each therapy period, the MNTs from 25 sites of the horses backs were assessed by pressure algometry (PA), and induced back movement was evaluated by flexion testing. Baseline MNTs were generally low, with means between 6.4 and 10.0 kg/cm². Significant changes in MNTs occurred nearly equally after both the PEMF and the placebo control treatments (at 5 of 25 or 6 of 25 sites, respectively) and without any regular pattern. Changes were evident, predominantly as decreased MNTs within and between treatment periods. Flexion testing revealed stiffness or avoidance in 19 of 20 horses. Results of the flexion testing showed an increased number of physiologic reactions at the end of both treatment periods compared with baseline values. The effect of PEMF on back pain and range of induced back movement could not be proven in this study. Although pretherapy values indicated the horses might have experienced back pain, all horses were still actively used in sport, and back pain might not have been severe enough to allow a significant effect to be demonstrated.     

Epidemiological survey,general blood biochemistry,and histological examination of slaughtered heavy horse breeds with hemorrhage in the adipose tissue in the crest of the neck

Fifty-four slaughtered horses were classified into groups having adipose tissue in the crest of the neck with or without hemorrhage (AH and NH groups, respectively). Blood biochemical tests (Alb, TP, T-bil, GOT, GPT, LDH, T-cho, and BUN) and an epidemiological survey (age, gender, weight, origin, breed, BCS, CNS, and hoof disease) were performed. T-bil tended to be high, while the other parameters were normal. Weight, BCS, and CNS were higher in the AH group (P<0.05). GOT was lower in the AH group (P<0.05). It was suspected that the horses in the AH group had lipomatosis. It was assumed that the adipose tissue of the horses in the AH group contained damaged capillaries, and inflammation was confirmed based on evidence of macrophages and lymphocytes.     

Evaluation of the genetic structure of sika deer (Cervus nippon) in Japan's Kanto and Tanzawa mountain areas,based on microsatellite markers

The browsing habits of sika deer (Cervus nippon) in Japan have caused serious ecological problems. Appropriate management of sika deer populations requires understanding the different genetic structures of local populations. In the present study, we used 10 microsatellite polymorphisms to explore the genetic structures of sika deer populations (162 individuals) living in the Kanto region. The expected heterozygosity of the Tanzawa mountain range population (Group I) was lower than that of the populations in the Kanto mountain areas (Group II). Our results suggest that moderate gene flow has occurred between the sika deer populations in the Kanto mountain areas (Group II), but not to or from the Tanzawa mountain range population (Group I). Also, genetic structure analysis showed that the Tanzawa population was separated from the other populations. This is probably attributable to a genetic bottleneck that developed in the Tanzawa sika deer population in the 1950s. However, we found that the Tanzawa population has since recovered from the bottleneck situation and now exhibits good genetic diversity. Our results show that it is essential to periodically evaluate the genetic structures of deer populations to develop conservation strategies appropriate to the specific structures of individual populations at any given time.