Full genome sequence and virulence analyses of the recent equine isolate of Japanese encephalitis virus

In the past 25 years, there has been only one case of Japanese encephalitis in horses in Japan. We determined the full genome sequence of the Japanese encephalitis virus (JEV) strain JEV/eq/Tottori/2003 isolated from an afflicted horse and also analyzed its virulence in mice. The sequence analysis showed that the genome of JEV/eq/Tottori/2003 is similar to that of genotype I, a dominant genotype of JEV presently circulating in Japan. Its neurovirulence, but not neuroinvasiveness, was still as high as it was for genotype III, thus indicating the necessity for continuation of a vaccination program of horses against JEV.     

Serosurveillance for Japanese encephalitis virus infection among equines in India

The seroprevalence of Japanese encephalitis virus (JEV) among equines was evaluated from January 2006 to December 2009 in 13 different states of India by hemagglutination inhibition (HI) test and virus neutralization test (VNT). Antibodies against JEV were detected in 327 out of 3,286 (10%) equines with a maximum prevalence reported in the state of Manipur (91.7%) followed by Gujarat (18.5%), Madhya Pradesh (14.4%), and Uttar Pradesh (11.6%). Evidence of JEV infection was observed in equines in Indore (Madhya Pradesh) where a 4-fold or higher rise in antibody titer was observed in 21 out of 34 horses in November 2007 to October 2006. In March 2008, seven of these horses had a subsequent 4-fold rise in JEV antibody titers while this titer decreased in nine animals. JEV-positive horse sera had a JEV/WNV (West Nile virus) ratio over 2.0 according to the HI and/or VNT. These results indicated that JEV is endemic among equines in India.     

Isolation and genetic analysis of Japanese encephalitis virus from a diseased horse in Japan

Japanese encephalitis (JE) developed in an unvaccinated half-bred horse kept in Tottori Prefecture, Japan. The animal showed ataxia with pyrexia and low appetite, and ultimately died. A viral strain was isolated from the cerebrum of the horse and was identified as JE virus (JEV) by RT-PCR using JEV specific primers. The isolated JEV was classified into genotype I by nucleotide sequence analysis of the viral envelope gene. We believe that this is the first report of the genotype I strain being isolated from a horse.     

Frequency of and risk factors for epistaxis associated with exercise-induced pulmonary hemorrhage in horses: 251,609 race starts (1992-1997)

OBJECTIVE: To determine the frequency of epistaxis during or after racing among racehorses and identify factors associated with development of epistaxis. DESIGN: Retrospective study. SAMPLE POPULATION: 247,564 Thoroughbred and 4,045 Anglo-Arab race starts. PROCEDURE: Race start information (breed, age, sex, racing distance, and race type) was obtained for Thoroughbred and Anglo-Arab horses racing in Japan Racing Association-sanctioned races between 1992 and 1997. All horses that raced were examined by a veterinarian within 30 minutes of the conclusion of the race; any horse that had blood at the nostrils was examined with an endoscope. If blood was observed in the trachea, epistaxis related to exercise-induced pulmonary hemorrhage (EIPH) was diagnosed. RESULTS: Epistaxis related to EIPH was identified following 369 race starts (0.15%). Frequency of EIPH-related epistaxis was significantly associated with race type, age, distance, and sex. Epistaxis was more common following steeplechase races than following flat races, in older horses than in horses that were 2 years old, following races < or =1,600 m long than following races between 1,601 and 2,000 m long, and in females than in sexually intact males. For horses that had an episode of epistaxis, the recurrence rate was 4.64%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that frequency of EIPH-related epistaxis in racehorses is associated with the horse's age and sex, the type of race, and the distance raced. The higher frequency in shorter races suggests that higher intensity exercise of shorter duration may increase the probability of EIPH.     

Nosological study of Borna disease virus infection in race horses.

To investigate the prevalence of diseases in the Borna disease virus (BDV) antibody positive race horses, we undertook seroepidemiological studies of BDV infection on 125 culled race horses in Hokkaido, Japan. The serological study disclosed the presence of antibodies only to BDV-p40 or -p24 in 19.2% (24/125) and 3.2% (4/125) of culled horses, respectively. Antibodies to both BDV-p40 and -p24 were found in 24.0% (30/125) of these horses. Of particular note was the finding that locomotorium disorders were detectable at a significantly higher rate in BDV antibody positive horses than that in the seronegative horses. These results imply that BDV infection may possibly contribute to an increase in the incidence rate of locomotorium disorders in race horses.     

Serosurveillance for Japanese encephalitis, Akabane, and Aino viruses for Thoroughbred horses in Korea

Recent global warming trends may have a significant impact on vector-borne viral diseases, possibly affecting vector population dynamics and disease transmission. This study measured levels of hemagglutination-inhibition (HI) antibodies against Japanese encephalitis virus (JEV) and neutralizing antibodies against Akabane virus (AKAV) and Aino virus (AINV) for Thoroughbred horses in Korea. Blood samples were collected from 989 racehorses in several provinces, between October 2005 and March 2007. Sera were tested using either an HI assay or a virus neutralization test. Approximately half (49.7%; 492/989) of the horses tested were antibody-positive for JEV. The HI titer against JEV was significantly correlated with racehorse age (p < 0.05). Horses with an HI antibody titer of 1:160 or higher accounted for 3.9% of the animals tested, indicating that vectors transmitting arthropod-borne viruses bit relatively few horses. In contrast, 3.8% (19/497) and 19.5% (97/497) of horse sera collected in March 2007 were positive against AKAV and AINV, respectively. The presence of antibodies against AKAV and AINV may indicate the multiplication of AKAV and AINV in these horses.     

Intranasal infection of Getah virus in experimental horses.

Aerosol transmission in equine Getah virus (GV) infection was examined by intranasal inoculation with 10(3.0) to 10(7.0) TCID50 of the MI-110 strain in 7 experimental horses. The establishment of intranasal infection of GV was confirmed in all these horses by detecting serum neutralizing antibody against the MI-110 strain. Horses inoculated with more than 10(4.0) TCID50 of the virus manifested mild pyrexia, eruptions, serous nasal discharge, lymphopenia or monocytosis. Viremia ranging from 10(1.0) to 10(3.5) TCID50/0.2 ml occurred in horses inoculated with 10(5.0) TCID50, or more. Virus recovery from the nasal cavity was observed only in horses inoculated with 10(7.0) TCID50, and the viral titers recorded were 10(3.0) TICD50/ml or less. From these results, it is assumed that GV disseminated from the nasal cavity of naturally infected horses, except for intranasal infection with a lot of the virus, is probably very low in titer. So it seems to be rare that GV in natural cycles is spread from horse to horse by aerosol transmission.     

Risk factors for atrial fibrillation during racing in slow-finishing horses

OBJECTIVE: To determine prevalence of atrial fibrillation (AF) immediately after racing among racehorses that finished well behind the winners and examine potential risk factors for AF in these horses. DESIGN: Case-control study. ANIMALS: 39,302 racehorses representing 404,090 race starts in races sanctioned by the Japan Racing Association between 1988 and 1997. PROCEDURE: Horses that finished > or = 4 (turf races) or 5 (dirt races) seconds behind the winner or that did not complete the race were examined for AF within 5 minutes after the race. Logistic regression and chi2 analyses were used to determine whether sex, age, race distance, race surface, year, or development of epistaxis was associated with development of AF. RESULTS: Estimated minimum frequency of AF was 0.03% (123 instances of AF following 404,090 race starts), and estimated minimum prevalence of AF among racehorses was 0.29% (115 horses with AF among 39,302 racehorses). Estimated frequency of AF among horses that finished slowly or did not finish was 1.39% (120 instances of AF among 8,639 examinations), and estimated prevalence of AF in horses that finished slowly was 1.23% (92 instances of AF among 7,500 horses) or 1.01% when only the first time a horse finished slowly was considered (76 instances of AF among 7,500 horses). Atrial fibrillation was paroxysmal in most horses. Among horses that finished slowly, 4-year-old and older horses and horses that raced on turf were more likely to develop AF. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the likelihood of AF among racehorses that finish slowly is related to age and racing surface.     

Effect of training location and time period on racehorse performance in New Zealand. 1. Descriptive analysis

AIM: To describe characteristics of Thoroughbred training stables in Matamata and in all other locations in New Zealand combined, over two 19-month time periods in 1996–1997 and 1998–1999, representing equal length periods immediately prior to and after the construction of a new training surface at the Matamata Racing Club.

METHODS: Retrospective records covering all horses training and racing in New Zealand during two 19-month time periods (1996–1997 and 1998–1999), covering 161 locations, were obtained from New Zealand Thoroughbred Racing (NZTR). Outcome variables included whether a horse was raced again in the 6 months following any start in the first 13 months of either time period, number of race starts for every horse, and finishing position. Summary measures with confidence intervals (CI) and unadjusted odds ratios (OR), measuring strength of associations for various factors, were computed.

RESULTS: The datasets contained information on 45,446 horses, 11,336 races, 5,110 trials and a total of 110,643 race starts. Horses trained at Matamata represented 8% (3,715) of the total horse datasets, and accounted for 11,977 race starts (10.8%). They were more likely to start in a race or trial in either time period and were 1.4 and 1.3 times as likely to finish first, second or third compared with horses trained at other locations in 1996–1997 and 1998–1999, respectively. A 6-month no-race period occurred for 9,306/12,584 (74%) horses that started at least once in the first 13 months of either time period. Horses trained at Matamata were less likely to have a 6-month no-race period than horses trained at other locations in both time periods. There was no effect of time period within each location on the probability of either a horse having a 6-month no-race period or of a race start being followed by a 6-month no-race period, but there was an overall effect of time and more 6-month no-race periods were observed in 1998–1999 relative to 1996–1997.

CONCLUSION: Summary statistics are presented for Thoroughbred racing in New Zealand over two 19-month time periods. Differences between the populations of horses trained in Matamata compared with those trained at other locations were attributed, in part, to the fact that many of the more successful racehorse trainers in the country have stables at Matamata. As a result, the population of horses in Matamata may not be representative of the racehorse population in New Zealand. Although more likely to win or place in both time periods, the magnitude of the advantage to horses in Matamata was reduced in 1998–1999 relative to 1996–1997, and this could be due, in part, to effects of the new track surface at Matamata. There was no evidence of a rise in risk of a 6-month no-race period following any race start in those horses trained in Matamata in 1998–1999 relative to either horses trained at other locations or to horses trained in Matamata during the earlier time period.     

A new modified live equine influenza virus vaccine: phenotypic stability, restricted spread and efficacy against heterologous virus challenge.

Flu Avert IN vaccine is a new, live attenuated virus vaccine for equine influenza. We tested this vaccine in vivo to ascertain 1) its safety and stability when subjected to serial horse to horse passage, 2) whether it spread spontaneously from horse to horse and 3) its ability to protect against heterologous equine influenza challenge viruses of epidemiological relevance. For the stability study, the vaccine was administered to 5 ponies. Nasal swabs were collected and pooled fluids administered directly to 4 successive groups of na?ve ponies by intranasal inoculation. Viruses isolated from the last group retained the vaccine's full attenuation phenotype, with no reversion to the wild-type virus phenotype or production of clinical influenza disease. The vaccine virus spread spontaneously to only 1 of 13 nonvaccinated horses/ponies when these were comingled with 39 vaccinates in the same field. For the heterologous protection study, a challenge model system was utilised in which vaccinated or na?ve control horses and ponies were exposed to the challenge virus by inhalation of virus-containing aerosols. Challenge viruses included influenza A/equine-2/Kentucky/98, a recent representative of the 'American' lineage of equine-2 influenza viruses; and A/equine-2/Saskatoon/90, representative of the 'Eurasian' lineage. Clinical signs among challenged animals were recorded daily using a standardised scoring protocol. With both challenge viruses, control animals reliably contracted clinical signs of influenza, whereas vaccinated animals were reliably protected from clinical disease. These results demonstrate that Flu Avert IN vaccine is safe and phenotypically stable, has low spontaneous transmissibility and is effective in protecting horses against challenge viruses representative of those in circulation worldwide.     

Effect of training location and time period on racehorse performance in New Zealand. 1. Descriptive analysis

AIM: To describe characteristics of Thoroughbred training stables in Matamata and in all other locations in New Zealand combined, over two 19-month time periods in 1996-1997 and 1998-1999, representing equal length periods immediately prior to and after the construction of a new training surface at the Matamata Racing Club. METHODS: Retrospective records covering all horses training and racing in New Zealand during two 19-month time periods (1996-1997 and 1998-1999), covering 161 locations, were obtained from New Zealand Thoroughbred Racing (NZTR). Outcome variables included whether a horse was raced again in the 6 months following any start in the first 13 months of either time period, number of race starts for every horse, and finishing position. Summary measures with confidence intervals (CI) and unadjusted odds ratios (OR), measuring strength of associations for various factors, were computed. RESULTS: The datasets contained information on 45,446 horses, 11,336 races, 5,110 trials and a total of 110,643 race starts. Horses trained at Matamata represented 8% (3,715) of the total horse datasets, and accounted for 11,977 race starts (10.8%). They were more likely to start in a race or trial in either time period and were 1.4 and 1.3 times as likely to finish first, second or third compared with horses trained at other locations in 1996-1997 and 1998-1999, respectively. A 6-month no-race period occurred for 9,306/12,584 (74%) horses that started at least once in the first 13 months of either time period. Horses trained at Matamata were less likely to have a 6-month no-race period than horses trained at other locations in both time periods. There was no effect of time period within each location on the probability of either a horse having a 6-month no-race period or of a race start being followed by a 6-month no-race period, but there was an overall effect of time and more 6-month no-race periods were observed in 1998-1999 relative to 1996-1997. CONCLUSION: Summary statistics are presented for Thoroughbred racing in New Zealand over two 19-month time periods. Differences between the populations of horses trained in Matamata compared with those trained at other locations were attributed, in part, to the fact that many of the more successful racehorse trainers in the country have stables at Matamata. As a result, the population of horses in Matamata may not be representative of the racehorse population in New Zealand. Although more likely to win or place in both time periods, the magnitude of the advantage to horses in Matamata was reduced in 1998-1999 relative to 1996-1997, and this could be due, in part, to effects of the new track surface at Matamata. There was no evidence of a rise in risk of a 6-month no-race period following any race start in those horses trained in Matamata in 1998-1999 relative to either horses trained at other locations or to horses trained in Matamata during the earlier time period.     

Hendra and Nipah virus infections.

The most important clinical and pathological manifestation of Hendra virus infection in horses and humans is that of severe interstitial pneumonia caused by viral infection of small blood vessels. The virus is also capable of causing nervous disease. Hendra virus is not contagious in horses and is spread by close contact with body fluids, such as froth from infected lungs. Diagnosis should be based on the laboratory examination of blood, lung, kidney, spleen, and, if nervous signs are present, also of the brain. Evidence of infection with the more recently identified and related Nipah virus was found in the brain of one horse in which there was inflammation of the meningeal blood vessels. Fruit bats, especially Pteropus s., have been incriminated as the natural and reservoir hosts of both Hendra and Nipah viruses.     

Effect of training location and time period on racehorse performance in New Zealand. 2. Multivariable analysis

AIM: To investigate training location (horses trained in Matamata vs those trained at all other venues in New Zealand), and time period (1996-1997 and 1998-1999), while controlling for other horse- and race- or trial-related factors, as a means of assessing the possible impact of construction of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance (number of starts, and failure to race within 6 months of any start). METHODS: Multivariable logistic regression and poisson analysis were used to analyse data derived using a retrospective cohort approach. Multivariable logistic regression was also used to analyse a case-control study. All data were derived from New Zealand Thoroughbred Racing (NZTR), records of race and trial results for racehorses trained in Matamata and other venues in New Zealand, covering two 19-month time periods (1996- 1997 and 1998-1999). Outcome variables included whether a horse started again in the 6 months following any start that occurred in the first 13 months of either time period, and a count of the total starts for every horse. RESULTS: Factors associated with increased risk of a start being followed by a 6-month no-race period included training location other than Matamata in comparison to horses trained in Matamata in the 1996-1997 time period, increasing age, 1998- 1999 over 1996-1997, starting in a trial rather than a race, placing fourth or worse in a start, softer track conditions, summer vs autumn, increasing cumulative exercise intensity in the 60 days prior to a start, and increasing race distance. Factors associated with an increase in the total number of starts included horses trained at Matamata in 1996-1997 compared with other time period-location combinations, younger age of horses at the time of a start, longer race distance, and an increasing proportion of starts in stakes races. CONCLUSIONS: Official race and trial results data provided a valuable resource for epidemiological studies of factors influencing racehorse performance. Results of analyses performed here provided little evidence of any adverse impact of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance.     

Cumulative racing-speed exercise distance cluster as a risk factor for fatal musculoskeletal injury in Thoroughbred racehorses in California

Thoroughbred racehorses which suffered a fatal musculoskeletal injury (FMI) while racing or race training at a California racetrack during 9 months of 1991 were studied to determine the importance of intensive, high-speed exercise schedules prior to injury. Seventy-seven horses which sustained an FMI while racing and 45 horses which sustained an FMI while race training were successfully matched by race or timed workout session with one control horse and included in the analyses. Race and timed workout (racing-speed exercise) histories were obtained for the case and control horses. Two-month cumulative, racing-speed cutoff distances were calculated from the control horse sample by two methods. Median racing-speed exercise frequencies and distances of the control horses were used to estimate age-specific (2, 3, 4 and ≥ 5 years), 2-month cumulative, racing-speed distances (Method 1). For the second method, the last race or timed workout for each control horse occurring just prior to, or on the date of injury for the matched case horse was identified. Cumulative racing-speed distances 2 months prior to these exercise events were determined for each control horse and used to estimate median age-specific (2, 3, 4 and ≥ 5 years), 2-month cumulative racing-speed distances (Method 2). The cumulative cutoff distances estimated from both methods were used to classify each matched pair according to the presence or absence of a 2-month cumulative, racing-speed distance which exceeded the age-appropriate cutoff distance (exercise distance cluster) within 6 months prior to injury. Manlel-Haenszel matched-pair odds ratios and 95% confidence limits were calculated separately for the racing and race-training fatal injuries. The relative risk for racing FMI was significantly greater for those horses which ran 2-month, cumulative racing and timed workout distances in excess of the cutoff values determined with Method 1 (relative risk (RR) = 3.0, 95% confidence interval (CI) = 1.2, 7.6) and Method 2 (RR = 7.2, 95% CI = 2.6, 20.6). The relative risk for race-training FMI was significantly greater for those horses which ran 2-month, cumulative racing and timed workout distances in excess of the cutoff values determined with Method 2 (RR = 3.4, 95% CI =1.0, 13.2).     

Effect of training location and time period on racehorse performance in New Zealand. 2. Multivariable analysis

AIM: To investigate training location (horses trained in Matamata vs those trained at all other venues in New Zealand), and time period (1996–1997 and 1998–1999), while controlling for other horse- and race- or trial-related factors, as a means of assessing the possible impact of construction of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance (number of starts, and failure to race within 6 months of any start).

METHODS: Multivariable logistic regression and poisson analysis were used to analyse data derived using a retrospective cohort approach. Multivariable logistic regression was also used to analyse a case-control study. All data were derived from New Zealand Thoroughbred Racing (NZTR), records of race and trial results for racehorses trained in Matamata and other venues in New Zealand, covering two 19-month time periods (1996–1997 and 1998–1999). Outcome variables included whether a horse started again in the 6 months following any start that occurred in the first 13 months of either time period, and a count of the total starts for every horse.

RESULTS: Factors associated with increased risk of a start being followed by a 6-month no-race period included training location other than Matamata in comparison to horses trained in Matamata in the 1996–1997 time period, increasing age, 1998–1999 over 1996–1997, starting in a trial rather than a race, placing fourth or worse in a start, softer track conditions, summer vs autumn, increasing cumulative exercise intensity in the 60 days prior to a start, and increasing race distance. Factors associated with an increase in the total number of starts included horses trained at Matamata in 1996–1997 compared with other time period-location combinations, younger age of horses at the time of a start, longer race distance, and an increasing proportion of starts in stakes races.

CONCLUSIONS: Official race and trial results data provided a valuable resource for epidemiological studies of factors influencing racehorse performance. Results of analyses performed here provided little evidence of any adverse impact of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance.     

Experimental infection of Japanese encephalitis virus in dogs

A previous serosurvey of Japanese encephalitis virus (JEV) among dogs suggested that dogs are well suited for use as sentinels for assessing the risk of JEV transmission to humans. To examine the clinical symptoms and duration of anti-JEV antibodies in dogs, three dogs were experimentally challenged with JEV. All JEV-infected dogs did not show any clinical signs or abnormal blood tests, except for C-reactive protein. Virus-neutralization titers rapidly increased and were maintained until 70 days postinfection, and neither the virus nor the viral genome was detected in blood. Thus, since dogs live in close proximity to humans as companion animals, they are well suited for use as sentinels for surveying the human risk of JEV infection.     

Pathogenicity for horses of original Sagiyama virus, a member of the Getah virus group

Sagiyama virus is a member of the Getah virus group. Its pathogenicity for horses was examined. All the horses infected with the original 4 strains of Sagiyama virus (M6/Mag 33, Mag 121, Mag 132 and Mag 258) developed pyrexia ranging from 39.0 to 40.0 degrees C. Other clinical signs, characterized by eruptions, edema in the hind legs, enlargement of the submandibular lymph node and mild leukopenia, were also manifested. Viremia occurred 1-4 days post-inoculation (p.i.). Virus was recovered from spleen, liver, lung and various lymph nodes of a horse autopsied on Day 4 p.i. The maximum titer of virus (10(6.0) TCID50 g-1) was detected in the inguinal lymph node. Seroconversion was demonstrated in all the infected horses on Day 5 p.i. These clinical signs and virological findings were similar to those of horses infected naturally. The results indicate that Sagiyama virus has pathogenicity for horses and is similar to that of Getah virus.     

Epizotiology and phylogeny of equine arteritis virus in hucul horses

The aim of the study was to determine the situation of equine arteritis virus (EAV) infections in hucul horses. A total of 176 horses (154 mares and 22 stallions) from the biggest hucul horse stud in Poland were tested. Antibodies against EAV were detected in 97 (55.1%) horses. The EAV seroprevalence among mares was 53.2% while in stallions - 68.2%. The percentage of positive mares increased with their age, thus amongst the mares of less than 2 years of age the percentage was 32.5%, while in the group of 3-5 years old increased to 59.4% and in the mares in the age of 6-10 years and older than 10 years 89.5% and 95% were seropositive, respectively. Among 11 seropositive stallions five were supposed to be shedders of EAV with their semen. It is likely that those persistently infected stallions were the reservoirs of the virus in the stud. Genetic studies using of ORF5 gene showed high homology between the viruses detected in the semen of those stallions what suggested lateral transmission between the stallions sharing the same stable. Persistent infection in an immature stallion, which has not yet been used for breeding, was established as a result of infection via respiratory route. Phylogenetic analysis confirmed that all hucul viruses shared the same ancestor and as most of EAV strains dominating in Polish horse population belonged to the European origin EAV subgroup (EU-1).     

Molecular phylogenetic analysis of bovine papular stomatitis viruses detected in Saga,Japan

In this study, we performed a molecular phylogenetic analysis of six bovine papular stomatitis virus (BPSV) field strains detected from Japanese beef calves kept on a farm in Saga prefecture, a southwest part of Japan, from 2017 to 2020. The phylogenetic analysis based on a partial B2L gene (554-nt) showed that these field strains were divided into two lineages, a lineage (A-lineage) constructed by a Saga strain and strains obtained from various regions of Japan and the world, and other lineage (B-lineage) constructed by five Saga strains and strains obtained from France, USA and Iwate prefecture (a north part of Japan). Furthermore, a Saga field strain named BPSV_SAGAbv2 and strains obtained from USA and Iwate prefecture belonged to a sub-lineage blanched from B-lineage. This is the first report elucidating molecular epidemiological characters of field BPSVs obtained from Saga prefecture. The existence of the multiple lineages was thought to be related to a history of calf introduction from various regions of Japan into Saga prefecture.