Population Statistics and Biological Traits of Endangered Kiso Horse

The objective of this study was to clarify the current status of endangered Kiso horse, population statistics and biological traits, in order to take a step for the conservation by scientific approach. We surveyed 125 Kiso horses (86.2% of the whole breed), analyzed the construction of the population, and calculated the coefficient of inbreeding and effective population size. Moreover, we confirmed coat color variations and the traditional traits of the Kiso horse, and measured their height at the withers and chest circumference to clarify their physical characteristics. The population pyramid of the horses was stationary or contractive, suggesting a reduction of the population in the near future. The effective population size of the horse (47.9) suggested that the diversity was much less than their census size, and the high coefficient of inbreeding, 0.11 ± 0.07 on average, suggested that the horses were surely inbred. The horses had only 4 coat colors; bay, dark bay, buckskin dun, and chestnut, and 116 horses (92.8%) were bayish color, suggesting the fixation in their coat color. Moreover, the majority of them had dorsal stripe (83 horses; 66.4%), and the average heights at withers(131.9 ± 4.4 cm) and chest circumference (167.1 ± 10.1 cm) were not significantly different between males and females.     

Genetic characterization of the endangered Kiso horse using 31 microsatellite DNAs

In order to contribute to conservation of the endangered Kiso horse, we clarified their genetic information using 31 microsatellite DNAs, and genotyped 125 horses, 83% of the existing breed. First, we clarified the current status of the horses. The horses were confirmed to have experienced rapid loss of population causing a bottleneck, and their effective population size was much smaller than their census size. Moreover, the number of alleles (6.3), observed heterozygosity (0.674), and expected heterozygosity (0.662) were in the same range as other endangered horses all over the world. Therefore, although their inbreeding level was not so severe (F(is): -0.017), the Kiso horse is surely one of the endangered. Second, we obtained genetic information of individuals. This information allowed us to understand the genetic distance of individuals, and might help in development of a reproductive strategy concerning the genetic distance between the mating pairs. Moreover, there appeared to be 4 subpopulations of Kiso horse, and this result was in good agreement with their historical background. Third, we confirmed that the parentage test for identification using the 31 microsatellite DNAs was highly reliable (probability of exclusion: 0.999999993). This identification increases the reliability of stud certification, and is also helpful for effective management. Understanding the genetic diversity within the population and the relationships among individuals is important to ensuring effective management for maintenance of genetic variation, and this study may help in conservation of the endangered Kiso horse.     

Isolation of virulent Rhodococcus equi from native Japanese horses

R. equi was isolated from soil samples obtained from the environment of seven native Japanese horse breeds (Hokkaido, Kiso, Noma, Misaki, Tokara, Miyako and Yonaguni) and from fecal samples collected from three native horse breeds (Hokkaido, Kiso and Misaki). Virulent R. equi at various levels (ranging from 0.5 to 12.9%) was isolated from the feces or soil environment of Hokkaido, Kiso and Misaki horses. Isolates were investigated both for the presence of 15- to 17-kDa antigens (virulence-associated protein antigens; VapA) by colony blotting, using the monoclonal antibody 10G5, and the gene of VapA by PCR. Plasmid DNAs extracted from positive isolates were digested with restriction endonucleases, and the digestion patterns of the plasmids of virulent isolates were divided into three types. Two of the three types (87-kb type II and 90-kb type I) had already been reported in Japanese isolates, and a new type (tentatively designated as 90-kb type II) had been found in isolates from Kiso horses. Six virulent R. equi isolates from the Hokkaido horses contained an 87-kb type II plasmid. Eight of 24 isolates from the Kiso horses contained an 87-kb type II plasmid, and the remaining 16 contained a 90-kb type II (a new type) plasmid. Two isolates from the Misaki horses contained a 90-kb type I plasmid. These results demonstrate the geographic difference in the distribution of virulence plasmids in R. equi isolates among native Japanese horses.     

Maximum permissible load for Kiso horses trotting over a short,straight course

This study aimed to determine the load‐bearing capacity of trotting Kiso horses using gait analysis. Ten Kiso horses with a height at withers of 128 cm were included. Their riders were fitted with a marker (70 mm in diameter placed on their chest) recorded by two digital DVD cameras while trotting along a short, straight course. In total, eight tests were performed for each horse: the first with a 70 kg load; six with randomly loaded weights ranging 80–130 kg; and then a final test again with a 70 kg load. Three‐dimensional movement of the marker was analyzed using a motion capture system. The time series of vertical displacement of the marker underwent spectrum analysis by the maximum entropy method, and the autocorrelation coefficient was calculated. The first two peaks of the autocorrelation were defined as symmetry and regularity, and their sum was defined as stability. Regularity in the 120 kg test (0.54) was lower than that in the first 70 kg test (0.61), and stability in the 120 kg test (1.31) was lower than that in the first 70 kg test (1.42). We concluded that the maximum permissible load for a trotting Kiso horse is < 120 kg, which represents 31% of its bodyweight.     

Dietary Capsaicin Does Not Alter Synovial Concentrations of Prostaglandin E2 or the Acute Phase Response in Aged Horses after Antigenic Challenge

Dietary capsaicin enhances disease resistance and immunity in various species. Because relatively little is known about the potential benefits of capsaicin when used on horses, this study was conducted to determine the effect of dietary capsaicin on measures of health in horses. Twelve horses were fed over 28 days a basal diet with three levels of dietary capsaicin: 0 mg (C), 50 mg (CAP50), or 100 mg (CAP100) per horse per day. Before feeding on day 0, horses were weighed, a blood sample taken, and a sample of synovial fluid from the left distal carpal joint was taken. Subsequent body weights and blood samples were obtained on days 7, 14, 21, and 28. On day 21, tetanus toxoid (TT) and an immunomodulator (EqStim) were given to each horse. On days 21 to 28, daily rectal temperature (RT) and blood samples were taken. On day 28, synovial fluid was obtained immediately after blood sampling and RT measurement. Synovial concentrations of prostaglandin E₂ did not differ among dietary treatments or between days 0 and 28. No effect of dietary capsaicin on serum immunoglobulin G subclass T or α₁-acid glycoprotein concentrations was observed. Serum haptoglobin was elevated (P < .0003) and RT increased (P < .05) after challenge with EqStim and TT; however, haptoglobin concentrations and RT did not differ due to diet. We conclude that the doses of dietary capsaicin fed to horses in this study had no beneficial effect on measures of joint health or the immune response in horses.     

Leptospira seroprevalence and associations between seropositivity,clinical disease and host factors in horses

Background

A cross-sectional study was carried out to determine the seroprevalence of different serovars of Leptospira spp. and their association with clinical disease and host factors in Swedish horses.

Methods

Sera from 2017 horses brought to equine clinics during 1997–98 were investigated. The sera were examined by microscopic agglutination test for the presence of antibodies against the following L. interrogans serovars: Bratislava strain Jez, Icterohaemorrhagiae strain Kantorowicz and Pomona strain Pomona and also L. kirschneri sv Grippotyphosa strain Duyster and L. borgpetersenii sv Sejroe strain M 84. Host factors, disease factors, season, pasture access and outdoor confinement variables were analysed with respect to seropositivity to sv Bratislava and Icterohaemorrhagiae. Multivariable logistic regression was used to model seropositivity to sv Bratislava and Icterohaemorrhagiae (seroprevalence > 8%).

Results

The seroprevalence, at a cut-off 1:100, were for sv Bratislava (16.6%), Icterohaemorrhagiae (8.3%), Sejroe (1.2%), Pomona (0.5%) and Grippotyphosa (0.4%). In the multivariable analysis, it was demonstrated that seroprevalence increased with age for sv Bratislava and Icterohaemorrhagiae. For sv Bratislava the seasons April – June and October – December and for sv Icterohaemorrhagiae October – December had higher seroprevalences than other seasons. Horses not used for racing had higher levels of seropositivity to sv Bratislava. Furthermore, horses with respiratory problems as well as horses with fatigue had higher levels of seropositivity to sv Bratislava. Ponies and coldbloods, and horses with access to pasture, had lower seroprevalence for sv Icterohaemorrhagiae. Healthy horses had lower seroprevalence for sv Icterohaemorrhagiae, than non-healthy horses.

Conclusion

There was no significant association between clinical signs and disease and positive titres to sv Bratislava (except for the association between respiratory problems and fatigue and seropositivity to sv Bratislava). The results suggest that horses with increasing age and exposed to factors associated with outdoor life had an increased seroprevalence for sv Bratislava, indicating that horses get infected from outdoor and/or are exposed to shedding from other horses (management dependent). For sv Icterohaemorrhagiae, management possibly plays a role as ponies and coldbloods as well as healthy horses had lower seroprevalence. Overall, the age of the horse should be taken into consideration when evaluating the titre as the average healthy horse has a higher titre than a young horse.     

Effects of pre-shipping marbofloxacin administration on fever and blood properties in healthy Thoroughbreds transported a long distance

The present study evaluated the effects of single-dose marbofloxacin in protecting horses against fever associated with transportation using 48 healthy Thoroughbreds. All horses were premedicated with interferon-α (0.5 U/kg, sublingually, every 24 hr) for 2 days before transportation and on the day of transportation. Horses were randomly assigned to receive marbofloxacin (2 mg/kg, IV, once; MRFX group), enrofloxacin (5 mg/kg, IV, once; ERFX group) or saline (0.9% NaCl) solution (10 ml, IV, once; control group) ≤1 hr before being transportation. Each group contained 16 horses (8 males, 8 females). Horses were transported 1,210 km using commercial vans over the course of approximately 26 hr. Clinical examinations and hematologic analyses were performed on all horses both before and after transportation. Post-transportation neutrophil to lymphocyte ratios were significantly lower in horses in the MRFX group compared with the control horses. The serum amyloid A levels were significantly lower in horses in the MRFX group and ERFX group compared with the control horses. Regarding the post-transportation rectal temperatures, fever was detected in 0 horses and 1 horse in the MRFX and ERFX groups, respectively, whereas fevers exceeding 39.1°C were detected in 2 horses in the control group. Additionally, the number of essential post-transportation treatments provided by veterinarians was reduced 3-fold in the MRFX and ERFX groups compared with the saline group. MRFX provided ERFX-like protection against fever associated with long-distance transportation, yielding significantly better protection than saline. Administration of MRFX just before transportation deserves a further study for efficacy in preventing horse fever associated with transportation.     

Target Group Segmentation in the Horse Buyers’ Market against the Background of Equestrian Experience

Whereas in former times horses were reserved primarily for people involved in agriculture, elite equestrians or the military, nowadays equestrian sport has become an activity for people with a wide variety of backgrounds. However, as more and more people become involved with equestrian sport today, the knowledge concerning animal husbandry in general is diminishing due to an alienation from agricultural themes in modern societies. As a consequence, this development affects both riding ability and the appraisal of horses, especially with respect to the purchase of horses. In order to analyse which factors influence purchase decisions in the horse market in conjunction with equestrian experience, 739 horse riders were surveyed on their purchase behaviour in this study. Using cluster analysis, a typology was generated that provides a differentiated picture of the preferences of the various rider groups. Three clusters were distinguished: the “amateurs”, the “experienced” and the “experts”. Taking personal horse riding proficiency into account, it could be concluded that especially the “amateur” group required objective criteria for the evaluation of a horse they are considering purchasing. Alongside “measureable” qualities, such as previous showing success or the level of training of the horse, also other attributes such as the simple handling of the horse should be taken into consideration. As particularly the “amateur” group in equestrian sport is increasing in numbers, it is therefore advisable when preparing a horse for sale to align oneself to the needs of this customer segment in order to ensure an effective and targeted marketing of horses.     

Effect of protective padding on forelimb intracompartmental muscle pressures in anesthetized horses

Wick catheters were used to measure intracompartmental muscle pressures (ICMP) within the long heads of the triceps brachii and extensor carpi radialis muscles of 8 horses maintained under halothane anesthesia while their breathing was controlled by intermittent positive-pressure ventilation. Blood gas, cardiac output, and blood pressure determinations were monitored to maintain a stable plane of anesthesia. The horses were positioned in left lateral recumbency and were placed sequentially on each of 4 contact surfaces for 1 hour. The 4 surfaces used for each horse were concrete, foam rubber, air dunnage bag, and a water mattress. Hematologic and biochemical determinations were made before and 24 hours after anesthesia. All horses recovered from the anesthesia. One horse had forelimb lameness for 36 hours after anesthesia, which was clinically diagnosed as a myoneuropathy. The ICMP values were markedly elevated in the muscle bellies of the lower limb of all horses. Supporting the horse on a water mattress caused the least dramatic pressure elevation and foam caused the most. The triceps muscle and, to a lesser extent, the extensor carpi radialis muscle of the lower limb are at risk of ischemia in anesthetized horses because the ICMP may exceed the critical closing pressure of 30 mm of Hg required for capillary blood flow.     

Evaluation of a positioning method for equine lateral stifle scintigrams

Background

The current lack of a standardized protocol for positioning of the gamma camera relative to the horse limb in a lateral stifle scintigram, and thus the reliance on subjective positioning, may be a cause of diagnostic error and inter-operator variability due to variations of the view angle. The aims of this study were to develop a reliable method to obtain a lateral scintigram of the equine stifle based on fixed anatomical landmarks and measure the resulting foot to gamma camera angle on sequential measurements of the same horse and of different horses

Methods

Technetium filled capsules were glued on the skin on sites adjacent to the origin of the medial and lateral femorotibial collateral ligaments in 22 horses using ultrasound guidance. A lateral view of the stifle was defined as the image where the two radioactive point sources were aligned vertically (point sources guided method). Five sequential lateral acquisitions (one to five) of the stifle with the point sources vertically aligned were acquired in each horse, and the angle between the mid-sagittal foot-axis and the vertical axis of the gamma camera (FC angle) was measured for each of these acquisitions

Results

For acquisition group one to five, the mean of the means FC angle was 91.6 ± 2° (2SD) and the coefficient of variation (COV) was 1.1%. In the 22 horses the 95% CI for the mean FC angles was 91.6° ± 12.1° (2SD) and the COV was 6.6%.

Conclusions

The use of point sources to guide gamma camera position results in less variation in the lateral scintigram than if the distal limb is used as guidance due to a difference in FC angle between horses. The point source guided positioning method is considered suitable as a reference standard method to obtain lateral scintigrams of the equine stifle, and it will be of value in clinical scintigraphy and research. The use of alignment of specifically located point sources may also be applied in other regions to standardize scintigraphic views.     

Dietary supplementation of a Saccharomyces cerevisiae fermentation product attenuates exercise-induced stress markers in young horses

Mitigation of exercise-induced stress is of key interest in determining ways to optimize performance horse health. To test the hypothesis that dietary supplementation of a Saccharomyces cerevisiae fermentation product would decrease markers of exercise-induced stress and inflammation in young horses, Quarter Horse yearlings (mean ± SD; 9 ± 1 mo) were randomly assigned to receive either no supplementation (CON; n = 8) or 21 g/d S. cerevisiae fermentation product (10.5 g/feeding twice daily; SCFP; n = 10) top-dressed on a basal diet of custom-formulated grain as well as ad libitum Coastal bermudagrass hay. After 8 wk of dietary treatments, horses underwent a 2-h submaximal exercise test (SET) on a free-stall mechanical exerciser. Serum was collected before dietary treatment supplementation (week 0), at week 8 pre-SET, and 0, 1, and 6 h post-SET and analyzed for concentrations of cortisol and serum amyloid A (SAA) by commercial enzyme-linked immunosorbent assay (ELISA) and for cytokine concentrations by commercial bead-based ELISA. Data were analyzed using linear models with repeated measures in SAS v9.4. From week 0 to 8 (pre-SET), serum cortisol decreased (P = 0.01) and SAA did not change, but neither were affected by diet. Serum concentrations of all cytokines decreased from week 0 to 8 (P ≤ 0.008), but granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor, and interleukin-8 (IL-8) decreased to a greater extent in CON than in SCFP horses (P ≤0.003). In response to the week 8 SET, serum cortisol increased in all horses (P < 0.0001) but returned to pre-SET levels by 1 h post-SET in horses receiving SCFP. At 6 h post-SET, cortisol concentrations in CON horses returned to pre-SET concentrations, whereas cortisol declined further in SCFP horses to below pre-SET levels (P = 0.0002) and lower than CON (P = 0.003) at that time point. SAA increased at 6 h post-SET in CON (P < 0.0001) but was unchanged through 6 h in SCFP horses. All cytokines except G-CSF increased in response to the SET (P < 0.0001) but showed differing response patterns. Concentrations of IL-1β, IL-6, and tumor necrosis factor-alpha were lesser (P ≤ 0.05), and concentrations of G-CSF and IL-18 tended to be lesser (P ≤ 0.09) in SCFP compared with CON horses throughout recovery from the SET. In summary, 8 wk of dietary supplementation with 21 g/d of SCFP may mitigate cellular stress following a single, prolonged submaximal exercise bout in young horses.     

Genetic Analysis of Three Lithuanian Native Horse Breeds

This study was the first to analyze the polymorphic characteristics of a wide range of genetic markers in three Lithuanian horse breeds. Genetic variation at seven red blood cell alloantigen (blood group) loci, 10 biochemical genetic loci and 16 microsatellites was examinated in 31 Zemaitukai, 30 large-type Zemaitukai and 24 Lithuanian Heavy Draught horse and compared to that of other domestic horse breeds. Genetic variability of the Zemaitukai breeds based upon blood groups and biochemical loci was higher than average for domestic horse breeds. For microsatellite loci, the variability of the large-type Zemaitukai and the Heavy Draught breeds also was high compared to the mean variation for domestic breeds; however, for the Zemaitukai breed, variation levels were very near the average for horses.     

Evaluation of Factors Contributing to Daily Within-Horse Variation of Plasma Alpha-Tocopherol Concentration

The current study was conducted to evaluate daily within-horse variation of plasma alpha-tocopherol concentration (TOC) (Experiment 1 [EXPT1]) and to determine the effect of a single meal on plasma TOC (Experiment 2 [EXPT2]). In EXPT1, venous jugular blood samples were collected from four pregnant (6–7 months of gestation), light horse mares (8.8 ± 2.9 years of age, mean ± SD; range, 5–11 years of age) between 7:30 AM and 8:30 AM, followed by eight hourly samples, and analyzed for plasma TOC. All horses were fed 5 kg dry matter (DM) grass hay 18 hours before sample collection and received no additional feed during the sampling period. Mean within-horse coefficient of variation for TOC was 3.5% ± 0.01%. In EXPT2, seven mature light horses, four mares and three geldings (4.9 ± 4.4 years of age, mean ± SD; range, 2–14 years of age), were randomly assigned to one of two treatments (fasted [FST; n = 7] or fed [FD; n = 7]), followed by reversal of treatments 24 hours later. At 7:30 AM, FD horses were fed 2.1 kg DM grain-mix concentrate (187 IU vitamin E/kg DM) plus 3.3 kg DM mixed-grass hay, whereas FST horses received no feed. Neither group received any feed for 18 hours before 7:30 AM. Venous jugular blood samples were collected at 10:30 AM from all horses and analyzed for plasma TOC. The mean (±SD) within-horse paired-treatment difference was not significant (0.05 ± 0.12 μg/mL). The results suggest that variation in within-horse plasma TOC is relatively small over an 8-hour period in fasting horses and is unaffected 3 hours after a hay grain meal.     

The relationship between respiratory exchange ratio, plasma lactate and muscle lactate concentrations in exercising horses using a valved gas collection system.

A valved gas collection system for horses was validated, then used to examine the relationship between the respiratory exchange ratio (RER), and plasma and muscle lactate in exercising horses. Four healthy Standardbred horses were trained to breathe through the apparatus while exercising on a treadmill. Comparisons of arterial blood gas tensions were made at 3 work levels for each horse, without (control), and with the gas collection system present. At the highest work level, the arterial oxygen tension (PaO2) was significantly lower (P < 0.05), and the arterial carbon dioxide tension (PaCO2) was significantly higher (P < 0.05), than control levels when the apparatus was present; however arterial oxygen content remained unchanged. The horses completed a standardized incremental treadmill test on 4 occasions to determine the repeatability of measurements of oxygen consumption (VO2), carbon dioxide production (VCO2), inspired minute ventilation (VI), respiratory exchange ratio (RER), ventilatory equivalent for oxygen (VI/VO2), tidal volume (VT), and ventilatory frequency (VF). All gas exchange and respiratory measurements showed good reproducibility with the mean coefficient of variation of the 4 horses ranging from 3.8 to 12%. We examined the relationship between 3 indices of energy metabolism in horses performing treadmill exercise: respiratory exchange ratio (RER), central venous plasma and muscle lactate concentrations. A relationship between RER and plasma lactate concentration was established. To compare muscle and plasma lactate concentrations, the horses completed a discontinuous exercise test without the gas collection apparatus present. Significant relationships (P < 0.05), between plasma lactate concentration and RER, and between plasma and muscle lactate concentration, were described for each horse. The valved gas collection system produced a measurable but tolerable degree of interference to respiration, and provided reproducible measurements of gas exchange and ventilatory measurements. It was concluded that measurements of both gas exchange and blood lactate may be used to indicate increased glycolytic activity within exercising skeletal muscle.     

The Kinetics of Hematopoiesis in the Light Horse I. The Lifespan of Peripheral Blood Cells in the Normal Horse

Three Standardbred horses were given 0.2 mg (1 mCi) of ⁷⁵selenomethionine intravenously and a second group of three were given 10 mCi of tritiated diisopropylfluorophosphate (0.5 mg) intravenously. Observations on labeled cells were continued for 250 days after radioselenium injection and 160 days after tritium injection.

The lifespan of erythrocytes using ⁷⁵selenmethionine was 155 ± 10 days and 148 ± 7.8 days using tritiated diisopropylfluorophosphate. There was no significant difference at the 10% level between the lifespans, using these labels. The uptake of radioselenium into erythrocytes reached a mean maximum of 11.5% at 82 ± 18.9 days after injection of the label. There was an elution of from 17.6% of the injected dose of tritium label down to 7.5% eight days after injection of this isotope. From this study both of these labels appear to be satisfactory for determining the erythrocyte lifespan of the horse.

The mean time of the curve of mean radioselenium activity of peripheral blood leukocytes for the three horses was 5.54 days and the lifespan of these cells was seven days. The mean lifespan of peripheral blood leukocytes of the three horses after in vivo labelling with tritiated diisopropylfluorophosphate was 17.3 hours. No specific labelling was found in bone marrow or peripheral blood cells at this level of tritium labelling by dipping emulsion autoradiography.

The mean lifespan of equine platelets for three horses using radioselenomethionine was 9.2 days and the mean lifespan of equine platelets using tritiated diisopropylfluorophosphate was 6.6 days in a group of three horses.

     

Temperature regulation in horses during exercise and recovery in a cool environment

Background

Clipping the winter coat in horses is done to improve heat dissipation during exercise and make grooming easier. It is often combined with blanketing to keep the horse warm. The aims of the present study were to investigate how clipping and the use of blankets affect thermoregulation during exercise and recovery in horses.

Methods

One Gotland pony, one New Forest pony, and one warm-blooded horse exercised one after the other on a 6450 m long track. The horses walked, trotted and cantered according to a predetermined scheme, which took about 50 minutes including three stops. The scheme was repeated on five consecutive days when horses were: 1) unclipped 2) unclipped + blanket during recovery, 3) left or right side clipped, 4) clipped, and 5) clipped + riding blanket + blanket during recovery. Heart rate (HR) was measured with telemetry, respiratory rate (RR) by counting flank contractions, skin temperatures by thermistor probes, and rectal temperature with a digital thermometer. Skin wetness (SW) was estimated by ocular inspection (dripping = 5, dry = 0).

Results

Mean outdoor temperature varied from -1.1 to - 8.7°C. HR increased progressively during exercise with no difference between treatments. Maximum RR was 77 ± 30 breaths/min (unclipped) and 49 ± 27 breaths/min (clipped). The lowest skin temperature was 17.5 ± 2.7°C in a hind leg during exercise, which increased to 34.5 ± 0.1°C during recovery. Rectal temperature was elevated during recovery in unclipped, but not in clipped horses and skin temperature at base of tail was elevated during recovery except in unclipped horses without blanket. Moisture after exercise scored 3.2 ± 0.8 in unclipped and zero in clipped horses.

Discussion and conclusion

Leg skin temperature initially dropped at onset of exercise in clipped horses, and then increased after about 30 minutes due to internal heat from the working muscles. These changes were not significant when clipped horses had riding blankets, whereas unclipped horses became overheated as judged from respiratory rate and elevated rectal temperature. Providing clipped horses with blankets dampened the changes in leg skin temperature during exercise.     

Detection of equine immunoglobulin-secreting cells by a plaque assay.

A protein A-hemolytic plaque assay was applied to detect immunoglobulin (Ig)-producing cells in horse peripheral blood, using pokeweed mitogen as a B lymphocyte activator. A maximum number of Ig-secreting cells was obtained when horse peripheral blood lymphocytes were cultured in a medium containing horse serum. The number of Ig-secreting cells in young horses (2 years old) was lower than that in adult horses (6 to 23 years old). In addition, the plaque formation was unchanged from blood samples kept at 4 degrees C for 24 hours, while blood samples kept for 72 hours did not yield plaques. These results indicate that the plaque assay is a reliable and useful method for detecting Ig-secreting cells in the peripheral blood of the horse.     

Salivary Cortisol Concentration in Exercised Thoroughbred Horses

Both physical activity and stress result in an increase in plasma cortisol level. The measurement of cortisol in plasma requires taking blood samples, which is stressful itself. Therefore, the aim of this study was to evaluate the use of saliva sampling for the determination of cortisol concentrations, indicating the intensity of exercise in horses during race training. Twelve Thoroughbred horses aged 2-3 years were examined during their speed training sessions. The horses galloped on the 1,200-m sand track at a speed of 14.4-15.3 m/s. Three saliva samples and three blood samples were collected from each horse. Both types of samples were taken when the horse was at rest, immediately after returning from the track and 30 minutes after the end of exercise. Blood lactic acid (LA) concentration was determined using the enzymatic cuvette test. The concentrations of cortisol in saliva and plasma samples were measured by enzyme immunoassay methods. Statistically significant correlations were found between salivary cortisol level determined 30 minutes after the end of exercise and blood LA concentration obtained immediately after exercise (P = .003) and between salivary and plasma cortisol levels measured 30 minutes after the end of training session (P = .015). The measurement of cortisol concentration in saliva samples taken from race horses 30 minutes after the end of exercise can be recommended for use in practice under field conditions to estimate the level of relative intensity of exercise in race horses.     

Molecular typing of VapA-positive Rhodococcus equi isolates from Jeju native horses, Korea

We recently demonstrated the presence of virulence-associated protein antigen (VapA)-positive Rhodococcus equi in Jeju Island, Korea. These bacteria contained one of two distinct plasmid types, a 90-kb type II plasmid, which has been found in isolates from the native Kiso horses of Japan, and a new variant, a 90-kb type V plasmid. However, the genotypic characters of the VapA-positive R. equi from Jeju native horses and their environments are poorly understood. Ninety-eight isolates from soil samples and 89 isolates from fecal samples were collected from five farms that breed or have bred Jeju native horses, and were tested for the presence of VapA by immunoblotting and PCR. Of the 98 soil isolates and 89 fecal isolates, seven and 13 were VapA-positive R. equi, respectively. In 2003, two Jeju foals died suddenly and were brought to the Faculty of Veterinary Medicine, Cheju National University, for postmortem examination. Pure cultures of R. equi were isolated from the lung lesions of both foals. Of the 16 clinical isolates, 14 were VapA-positive R. equi. Of the 34 VapA-positive clinical and environmental isolates, 16 contained the 90-kb type II plasmid and 18 contained a 90-kb type V plasmid. Pulsed-field gel electrophoresis (PFGE) patterns of the VapA-positive isolates from Jeju horses and Kiso horses, containing the 90-kb type II plasmid, were compared and formed two distinct groups. Furthermore, 18 virulent isolates containing the 90-kb type V plasmid formed two distinct PFGE groups (of 16 and two isolates). These results demonstrate that two virulence plasmid types are widespread in R. equi in Jeju native horses. However, there is little diversity in the PFGE patterns of virulent isolates, suggesting the clonal spread of virulent R. equi. The PFGE pattern of the virulent R. equi isolates from Jeju native horses in Korea is not identical to those of Kiso native horses in Japan.     

Glycated Hemoglobin is an Indicator of Blood Glucose Status in Horses: Preliminary Study

Glycated hemoglobin (HbG) concentration is a retrospective measure of mean blood glucose level and is not affected by recent stresses, food ingestion, or exercise. Although HbG has been determined in various wild and domestic animals such as kestrels, mankhor, mouflon, aoudad, deer, goat, sheep, dog, camel, ostrich, and horse, there is no information about diagnostic values of HbG as an indicator of blood glucose status in horses. The purposes of this study were to determine normal value of HbG in Iranian crossbred horses and to investigate its relation to fasting plasma glucose. Blood samples were collected from jugular veins of 193 clinically healthy adult crossbred horses (102 males and 91 females). After separation and washing of red blood cells, hemolysate was prepared and subjected to weak cation exchange chromatography for determination of HbG. Glucose was measured in fasting plasma samples. HbG percent (HbG%) in the studied horses was 3.21 ± 0.56 in males and 3.34 ± 0.72 in females. Fasting plasma glucoses were 81.3 ± 7.6 and 84.2 ± 14.5 mg/dL in males and females, respectively. HbG% and plasma glucose were highly correlated (r = 0.81, P < .01). We concluded that HbG% can be considered as a good indicator of blood glucose status in the horse. Two horses with abnormally higher HbG% were found in this study showing persistent high fasting plasma glucose. We concluded that HbG determination could be a more reliable indicator of basal blood glucose concentrations in horses.