Yoshiro Endo

Genetic and serological surveillance for non-primate hepacivirus in horses in Japan.

Non-primate hepacivirus (NPHV) is a recently discovered homolog of the hepatitis C virus in horses. The frequency and distribution of NPHV infections among horses in Japan is unknown. In this study, serum samples from 453 horses across Japan were screened for NPHV RNA using real-time RT-PCR and anti-nonstructural 3 protein (NS3) antibodies using the Gaussia luciferase immunoprecipitation system assay. In order to monitor the course of NPHV infection in horses, we examined 31 stored samples (9 adult horses and 22 young horses) obtained one year ago and compared the results to the recent data. Stored sera from 7 mare-foal pairs were also examined. The NS3 region sequences of 14 NPHV strains from NPHV RNA positive serum samples were determined and analyzed phylogenically. Of the 453 serum samples tested, 33.55% were positive for anti-NS3 antibody and 13.68% were positive for NPHV RNA. We found a higher rate of NPHV RNA detection in serum obtained from young horses (1-2 years of age) than that of adults, in two geographically distinct areas. We observed higher variation in the course of infection over one year in young horses than in adult horses. The foals were infected with NPHV after the weaning period. Phylogenic analysis revealed that while NPHV NS3 genes isolated in Japan clustered with sequences previously classified as NPHV, but the genetic diversity of the Japanese NPHV strains we detected was not correlated with their geographic origin. In conclusion, Japanese horses exhibit a high prevalence of NPHV. Young age appears to be a risk factor for such viral infection in Japan, although the infectious route was not determined.

Ultrasonographic Evaluation of Equine Fetal Growth Throughout Gestation in Normal Mares Using a Convex Transducer

ABSTRACT It has not been common to perform regular ultrasound examination of the fetus in equine practice, due to the increasing volume of the uterus caused by fetal development. The convex three-dimensional transducer is bulb-shaped and is able to observe wide areas. In addition, its operation is simple, making it easy to create appropriate angles for various indices using a transrectal approach. The aim of this study was to measure Thoroughbred fetal growth indices throughout gestation using a convex transducer and to clarify the detectable period of some indices for clinical use. We demonstrated changes in fetal indices, such as crown rump length (CRL), fetal heart rate (FHR), fetal eye and kidney and the combined thickness of uterus and placenta (CTUP). CTUP increased from 30 weeks of gestation, and FHR peaked at 8 weeks and then decreased to term. CRL could be observed until 13 weeks due to its wide angle, longer than in previous reports. Fetal eye and kidney could be observed from 10 and 28 weeks, respectively, and these increased with pregnancy progress. The present results showed the advantage of transrectal examination using a convex transducer for evaluation of normal fetal development. Although ultrasonographic examination in mid- to late-gestation is not common in equine reproductive practice, our comprehensive results would be a useful basis for equine pregnancy examination.

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.

Effects of ozonated autohemotherapy on the antioxidant capacity of Thoroughbred horses

The performance of horses undergoing regular intense exercise is adversely affected by oxidative stress. Thus, it is important to increase antioxidant production in horses in order to reduce oxidative stress. Ozonated autohemotherapy (OAHT) reportedly promotes antioxidant production. This study aimed to evaluate the effects of OAHT on antioxidant capacity. Ten Thoroughbred horses were used in this study. After the OAHT, we collected serum samples and measured biological antioxidant potential (BAP). We found that BAP began to increase after the OAHT and was significantly higher in the OAHT group than at 3 (P<0.01) and 7 days (P<0.05) after OAHT than in the control group at 3 and 7 days after starting collection of blood samples. Therefore, it was shown that OAHT improved the antioxidant capacity of the horses.

Oxidative stress markers in Thoroughbred horses after castration surgery under inhalation anesthesia.

ABSTRACT Oxidative stress has been reported to occur during surgery. It is important to reduce intraoperative oxidative stress to improve the postoperative prognosis. However, there are no reports regarding oxidative stress related to surgery in horses. In the present study, we measured pre and postsurgical diacron-reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP); the oxidative stress index (OSI) was then calculated (OSI=d-ROMs/BAP × 100). d-ROMs were not significantly different between the pre and postsurgical periods. However, BAP significantly decreased after surgery (P=0.02), and OSI significantly increased after surgery (P=0.02). Based on these results, it suggested that castration surgery under inhalation anesthesia decreases the antioxidant potential and causes oxidative stress in horses.

The cross‐sectional area changes in digital flexor tendons and suspensory ligament in foals by ultrasonographic examination

REASONS FOR PERFORMING STUDY Flexural deformities are common conditions of growing horses and are suggested to have a relationship with the contraction of musculotendinous units. However, limited studies have documented the changes in each tendon and ligament in the metacarpal region with age. OBJECTIVES To investigate the changes in the cross-sectional area (CSA) of each tendon and ligament in the metacarpal region with age by ultrasonographic examination. STUDY DESIGN Longitudinal study of foals from Day 1 to age 24 months. METHODS The CSA of the superficial digital flexor tendon, deep digital flexor tendon, accessory ligament of the deep digital flexor tendon and suspensory ligament was measured by ultrasonographic examination at monthly intervals from Day 1 to age 24 months in 7 Thoroughbred foals. RESULTS Changes in superficial digital flexor CSA in all regions were larger than those of other structures from 10 months to 15 months. The suspensory ligament CSA was significantly larger than those of other structures on Day 1 in both the region of suspensory origin (RSO) and region of suspensory body (RSBO). This condition continued until 2 months in the RSO and until 5 months in the RSBO. The changes in deep digital flexor CSA were larger than those of other structures from 2 to 5 months in both the RSO and RSBO. CONCLUSIONS The rate of change in each structure varies with age. Thus, the functional adaptation with age that takes place may differ among structures because the primary function of each structure differs.

Comparison of the Occurrence of Transportation-associated Fever in 2 Years Old Thoroughbreds before and after Introduction of Prophylactic Marbofloxacin Administration

ABSTRACT In order to reveal the preventive effect of marbofloxacin (MRFX) administration just before transportation, we compared the occurrence of transportation-associated fever before and after introduction of MRFX administration. After the introduction of prophylactic MRFX administration, the rectal temperatures of horses after transportation were significantly lower than before the introduction of MRFX administration (P<0.01) and the number of febrile horses was significantly lower than before the introduction of MRFX administration (P<0.01). In conclusion, these results show that prophylactic MRFX administration just before transportation is clinically effective at preventing transportation-associated fever.

Comparison of physical body growth and metabolic and reproductive endocrine functions between north and south climates of Japan in trained Thoroughbred yearling horses

ABSTRACT This study aimed to compare body growth, metabolic, and reproductive hormonal changes in trained Thoroughbred yearling horses under different climate conditions with and without light supplementation (LS). Thoroughbred yearlings raised at research centers of the Japan Racing Association in Hokkaido (north) or Miyazaki (south) were divided into control and LS groups. In the LS groups, 44 colts and 47 fillies from Hokkaido and 11 colts and 11 fillies from Miyazaki were exposed to LS with an extended photoperiod of 14.5 hr of daylight and 9.5 hr of darkness. One week before and once a month after LS, circulating total thyroxine (T4), insulin-like growth factor-I (IGF-1), prolactin (PRL), cortisol, and progesterone (P4) concentrations were measured by radioimmunoassay and fluoroimmunoassay, respectively. Growth parameters, including body weight, height, girth, and cannon bone circumferences, were measured monthly. Hair coat (HC) condition was scored. Under natural conditions, the T4 concentrations of Hokkaido yearlings tended to be higher, whereas the IGF-1 (colt) and PRL levels were significantly lower than those of yearlings in Miyazaki. Growth parameters and HC scores were lower in Hokkaido yearlings. With LS, the PRL and P4 concentrations in Hokkaido and Miyazaki were higher, and the first ovarian activity tended to be earlier than in the controls. Only LS Hokkaido yearlings showed significantly higher HC scores than the controls. Comparing the different climates among the LS yearlings, the levels of PRL and P4 and the HC scores in Hokkaido yearlings increased and reached levels similar to those in Miyazaki yearlings. The body weight and girth increment percentages of Hokkaido yearlings in January dramatically decreased and then eventually increased to levels similar to those of Miyazaki yearlings. This suggested that yearlings in naturally colder Hokkaido exhibit higher basal metabolism to maintain homeostasis. However, providing LS may help to improve growth and early development of reproductive function in Hokkaido yearlings to levels equal to those of Miyazaki horses.

Effects of low-dose G-CSF formulation on hematology in healthy horses after long-distance transportation

The present study evaluated the effects of single-dose filgrastim on hematology in 16 healthy horses after long-distance transportation. Horses were assigned to receive filgrastim (0.23 µg/kg, SC, once; G-CSF group; n=8) or saline (0.9% NaCl) solution (0.3 ml, SC, once; control group; n=8) ≤ 1 hr before transportation. Horses were transported 2,530 km using commercial vans over the course of approximately 44 hr. Clinical examinations and hematologic analyses were performed on all horses before and after transportation. Because the post-transportation white blood cell counts and bacillary neutrophil to segmented neutrophil ratio were significantly higher in the G-CSF group, filgrastim may have promoted the mobilization of neutrophils from marrow. Filgrastim deserves a further study for efficacy in preventing horse shipping fever.

A study of the distribution of color Doppler flows in the superficial digital flexor tendon of young Thoroughbreds during their training periods

ABSTRACT Aim of this study was to evaluate the relationships of exercise and tendon injury with Doppler flows appearing in the superficial digital flexor tendon (SDFT) of young Thoroughbreds during training periods. The forelimb SDFTs of 24 one- to two-year-old Thoroughbreds clinically free of any orthopaedic disorders were evaluated using grey-scale (GS) and color Doppler (CD) images during two training periods between December 2013 to April 2015. Twelve horses per year were examined in December, February, and April in training periods that began in September and ended in April. The SDFT was evaluated in 3 longitudinal images of equal lengths (labelled 1, 2, 3 in order from proximal to distal), and 6 transversal images separated by equal lengths (labelled 1A, 1B, 2A, 2B, 3A and 3B in order from proximal to distal) of the metacarpus using both GS and CD. The running (canter and gallop) distance for 1 month before the date of the ultrasonographic examinations was increased in December, February, and April in both of the two training periods. CD flows defined as rhythmically blinking or pulsatory colored signals were found in 56 of 864 (6.4%) transversal CD images, in 28, 12, 13, and 3 images of 1A, 1B, 2A and 2B, respectively, and in 7, 14, and 35 images captured in December, February, and April, respectively. There were no longitudinal or transversal GS images indicating injury in the SDFTs in either of the two training periods. The increase of CD flows in the proximal regions of the SDFT are possibly related to the increase of the running distance during the training periods of the one- to two-year-old Thoroughbreds. Because no injury was diagnosed in the SDFTs by GS images during the training periods, the increase of CD flows in the proximal parts of SDFT is not necessarily predictive of tendon injury in the near future during the training period of young Thoroughbreds.