Investigation of an outbreak of encephalomyelitis caused by West Nile virus in 136 horses

OBJECTIVE: To describe an outbreak of encephalomyelitis caused by West Nile virus (WNV) in horses in northern Indiana. DESIGN: Case series. ANIMALS: 170 horses. PROCEDURES: Horses with clinical signs suggestive of encephalomyelitis caused by WNV were examined. Date, age, sex, breed, and survival status were recorded. Serum samples were tested for anti-WNV antibodies, and virus isolation was attempted from samples of brain tissue. Climate data from local weather recording stations were collected. An epidemic curve was constructed, and case fatality rate was calculated. RESULTS: The most common clinical signs were ataxia, hind limb paresis, and muscle tremors and fasciculations. Eight horses had been vaccinated against WNV from 2 to 21 days prior to the appearance of clinical signs. West Nile virus was isolated from brain tissue of 2 nonvaccinated horses, and anti-WNV IgM antibodies were detected in 132 nonvaccinated horses; in 2 other nonvaccinated horses, anti-WNV antibodies were detected and WNV was also isolated from brain tissue. Thirty-one (22.8%) horses died or were euthanatized. The peak of the outbreak occurred on September 6, 2002. Ambient temperatures were significantly lower after the peak of the outbreak, compared with prior to the peak. CONCLUSIONS AND CLINICAL RELEVANCE: The peak risk period for encephalomyelitis caused by WNV in northern Indiana was mid-August to mid-September. Reduction in cases coincided with decreasing ambient temperatures. Because of a substantial case fatality rate, owners of horses in northern Indiana should have their horses fully protected by vaccination against WNV before June. In other regions of the United States with a defined mosquito breeding season, vaccination of previously nonvaccinated horses should commence at least 4 months before the anticipated peak in seasonal mosquito numbers, and for previously vaccinated horses, vaccine should be administered no later than 2 months before this time.     

Equine encephalomyelitis outbreak caused by a genetic lineage 2 West Nile virus in Hungary

Background: The spread of lineage 2 West Nile virus (WNV) from sub‐Saharan regions to Europe and the unpredictable change in pathogenicity indicate a potential public and veterinary health threat and requires scientific awareness. Objectives: To describe the results of clinical and virological investigations of the 1st outbreak of a genetic lineage 2 WNV encephalomyelitis in horses. Animals: Seventeen horses with neurologic signs. Methods: Information regarding signalment, clinical signs, and outcome was obtained for each animal. Serology was performed in 15 cases, clinicopathological examination in 7 cases, and cerebrospinal fluid was collected from 2 horses. Histopathology was carried out in 4 horses, 2 of which were assessed for the presence of WNV in their nervous system. Results: WNV neutralizing antibody titers were between 10 and 270 (median, 90) and the results of other serological assays were in agreement with those of the plaque reduction neutralization test. Common signs included ataxia, weakness, asymmetric gait, muscle tremors, hypersensitivity, cranial nerve deficits, and recumbency. Twelve animals survived. Amplicons derived from the infection‐positive specimens allowed molecular characterization of the viral strain. Conclusions and Clinical Importance: From our results, we conclude that this outbreak was caused by a lineage 2 WNV strain, even though such strains often are considered nonpathogenic. Neurological signs and survival rates were similar to those reported for lineage 1 virus infections. The disease occurrence was not geographically limited as had been the typical case during European outbreaks; this report describes a substantial northwestern spread of the pathogen.     

Characteristics of an outbreak of West Nile virus encephalomyelitis in a previously uninfected population of horses

Equine West Nile virus (WNV) encephalomyelitis cases - based on clinical signs and ELISA serology test results - reported to Texas disease control authorities during 2002 were analyzed to provide insights into the epidemiology of the disease within a previously disease-free population. The epidemic occurred between June 27 and December 17 (peaking in early October) and 1,698 cases were reported. Three distinct epidemic phases were identified, occurring mostly in southeast, northwest and then central Texas. Significant (P<0.05) disease clusters were identified in northwest and northern Texas. Most (91.1%) cases had no recent travel history, and most (68.9%) cases had not been vaccinated within the previous 12 months. One-third of cases did not survive, 71.2% of which were euthanatized. The most commonly reported presenting signs included ataxia (69%), abnormal gait (52%), muscle fasciculations (49%), depression (32%) and recumbency (28%). Vaccination status, ataxia, falling down, recumbency and lip droop best explained the risk of not surviving WNV disease. Results suggest that the peak risk period for encephalomyelitis caused by WNV may vary substantially among regions within Texas. Recumbent horses have a poor prognosis for survival. Vaccines, even if not administered sufficiently in advance of WNV infection within a district, may reduce the risk of death by at least 44%.     

Immunologic responses to West Nile virus in vaccinated and clinically affected horses

OBJECTIVE: To compare neutralizing antibody response between horses vaccinated against West Nile virus (WNV) and horses that survived naturally occurring infection. DESIGN: Cross-sectional observational study. ANIMALS: 187 horses vaccinated with a killed WNV vaccine and 37 horses with confirmed clinical WNV infection. PROCEDURE: Serum was collected from vaccinated horses prior to and 4 to 6 weeks after completion of an initial vaccination series (2 doses) and 5 to 7 months later. Serum was collected from affected horses 4 to 6 weeks after laboratory diagnosis of infection and 5 to 7 months after the first sample was obtained. The IgM capture ELISA, plaque reduction neutralization test (PRNT), and microtiter virus neutralization test were used. RESULTS: All affected horses had PRNT titers > or = 1:100 at 4 to 6 weeks after onset of disease, and 90% (18/20) maintained this titer for 5 to 7 months. After the second vaccination, 67% of vaccinated horses had PRNT titers > or = 1:100 and 14% had titers < 1:10. Five to 7 months later, 33% (28/84) of vaccinated horses had PRNT titers > or = 1:100, whereas 29% (24/84) had titers < 1:10. Vaccinated and clinically affected horses' end point titers had decreased by 5 to 7 months after vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: A portion of horses vaccinated against WNV may respond poorly. Vaccination every 6 months may be indicated in certain horses and in areas of high vector activity. Other preventative methods such as mosquito control are warranted to prevent WNV infection in horses.     

Pathogenicity of West Nile virus in chickens

In the fall of 1999, West Nile virus (WNV) was isolated for the first time in the Western Hemisphere during an outbreak of neurologic disease in humans, horses, and wild and zoo birds in the northeastern United States. Chickens are a potential reservoir for WNV, and little is known about the pathogenicity of WNV in domestic chickens. Seven-week-old chickens derived from a specific-pathogen-free flock were inoculated subcutaneously with 1.8 x 10(3) 50% tissue culture infectious dose of a crow isolate of WNV in order to observe clinical signs and evaluate the viremic phase, gross and microscopic lesions, contact transmission, and immunologic response. There were no observable clinical signs in the WNV-inoculated chickens during the 21-day observation period. However, histopathologic examination of tissues revealed myocardial necrosis, nephritis, and pneumonitis at 5 and 10 days postinoculation (DPI); moderate to severe nonsuppurative encephalitis also was observed in brain tissue from one of four inoculated birds examined at 21 DPI. WNV was recovered from blood plasma for up to 8 DPI. Virus titers as high as 10(5)/ml in plasma were observed at 4 DPI. Fecal shedding of virus was detected in cloacal swabs on 4 and 5 DPI only. The WNV also was isolated from myocardium, spleen, kidney, lung, and intestine collected from chickens euthanatized at 3, 5, and 10 DPI. No virus was isolated from inoculated chickens after 10 DPI. Antibodies specific to WNV were detected in inoculated chickens as early as 5 DPI by the plaque reduction neutralization test and 7 DPI by the indirect fluorescent antibody test. Chickens placed in contact with inoculated chickens at 1 DPI lacked WNV-specific antibodies, and no WNV was isolated from their blood plasma or cloacal swabs throughout the 21 days of the experiment.     

Severe acute rhabdomyolysis associated with Streptococcus equi infection in four horses

Four Quarter Horses (9 months to 7 years of age) with submandibular lymphadenopathy and firm muscles (palpation of which elicited signs of pain) were evaluated; in general, the horses had a stiff gait, and 3 horses became recumbent. Streptococcus equi was cultured from aspirates of lymph nodes or samples of purulent material collected from the auditory tube diverticula. Once the horses were recumbent, their condition deteriorated rapidly despite aggressive antimicrobial and antiinflammatory treatment, necessitating euthanasia within 24 to 48 hours. One horse did not become recumbent and recovered completely. Among the 4 horses, common clinicopathologic findings included neutrophilia, hyperfibrinogenemia, and high serum activities of creatine kinase and aspartate aminotransferase. Necropsies of the 3 euthanatized horses revealed large, pale areas most prominent in the semimembranosus, semitendinosus, sublumbar, and gluteal muscles that were characterized histologically by severe acute myonecrosis and macrophage infiltration of necrotic myofibers. Streptococcus equi was identified in sections of affected muscle by use of immunofluorescent stains for Lancefield group C carbohydrate and S. equi M protein. In the 4 horses of this report, acute severe rhabdomyolysis without clinical evidence of muscle atrophy or infarction was associated with S. equi infection; rhabdomyolysis was attributed to either an inflammatory cascade resembling streptococcal toxic shock or potentially direct toxic effects of S. equi within muscle tissue.     

Congestive heart failure in horses: 14 cases (1984-2001)

OBJECTIVE: To identify clinical signs, underlying cardiac conditions, echocardiographic findings, and prognosis for horses with congestive heart failure. DESIGN: Retrospective study. ANIMALS: 14 horses. PROCEDURE: Signalment; history; clinical signs; clinicopathologic, echocardiographic, and radiographic findings; treatment; and outcome were determined by reviewing medical records. RESULTS: All 14 horses were examined because of a heart murmur; tachycardia was identified in all 14. Twelve horses had echocardiographic evidence of enlargement of 1 or more chambers of the heart. Other common clinical findings included jugular distention or pulsation, crackles, cough, tachypnea, and ventral edema. Nine horses had signs consistent with heart failure for > 6 days. Underlying causes for heart failure included congenital defects, traumatic vascular rupture, pericarditis, pulmonary hypertension secondary to heaves, and valvular dysplasia. Seven horses were euthanatized after diagnosis of heart failure; 5 were discharged but were euthanatized or died of complications of heart disease within 1 year after discharge. The remaining 2 horses were discharged but lost to follow-up. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that congestive heart failure is rare in horses. A loud heart murmur accompanied by either jugular distention or pulsation, tachycardia, respiratory abnormalities (crackles, cough, tachypnea), and ventral edema were the most common clinical signs. Echocardiography was useful in determining the underlying cause in affected horses. The long-term prognosis for horses with congestive heart failure was grave.     

Evaluation of the efficacy provided by a Recombinant Canarypox-Vectored Equine West Nile Virus vaccine against an experimental West Nile Virus intrathecal challenge in horses

Efficacy of the Recombitek Equine West Nile Virus (WNV) vaccine was evaluated against a WNV intrathecal challenge model that results in WNV-induced clinical disease. Ten vaccinated (twice at days 0 and 35) and 10 control horses were challenged 2 weeks after administration of the second vaccine with a virulent WNV by intrathecal administration. After the challenge, eight of 10 controls developed clinical signs of encephalomyelitis whereas one vaccinate exhibited muscle fasciculation only once. Nine controls and one vaccinate developed a fever. Histopathology revealed mild to moderate nonsuppurative encephalitis in eight controls and one vaccinate. None of the vaccinates and all of the controls developed WNV viremia after challenge. All vaccinated horses developed antibodies to WNV after vaccination. These and results of previous studies demonstrate efficacy of the Recombitek WNV vaccine against WNV-induced clinical disease and natural challenge with WNV-infected mosquitoes.     

Evaluation of an outbreak of West Nile virus infection in horses: 569 cases (2002)

OBJECTIVE: To characterize an outbreak of West Nile virus (WNV) infection in horses in North Dakota in 2002, evaluate vaccine effectiveness, and determine horse characteristics and clinical signs associated with infection. DESIGN: Retrospective study. ANIMALS: 569 horses. PROCEDURE: Data were obtained from veterinary laboratory records, and a questionnaire was mailed to veterinarians of affected horses. RESULTS: Affected horses were defined as horses with typical clinical signs and seroconversion or positive results of virus isolation; affected horses were detected in 52 of the 53 counties and concentrated in the eastern and northeastern regions of the state. Among affected horses, 27% (n = 152) were vaccinated against WNV, 54% (309) were not, and 19% (108) had unknown vaccination status; 61 % (345) recovered, 22% (126) died, and 17% (98) had unknown outcome. The odds of death among nonvaccinated horses were 3 and 16 times the odds among horses that received only 1 or 2 doses of vaccine and horses that were vaccinated according to manufacturer's recommendations, respectively. Horses with recumbency, caudal paresis, and age > 5 years had higher odds of death, whereas horses with incoordination had lower odds of death, compared with affected horses without these characteristics. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination appears to have beneficial effects regarding infection and death caused by WNV.     

Botulism associated with feeding alfalfa hay to horses

Botulism was believed to be the cause of progressive symmetric myasthenia in 8 horses on a farm in North Carolina. One horse was found dead, 6 were euthanatized after becoming recumbent, and 1 affected horse recovered. Cecal and colonic contents of 2 horses were determined to contain Clostridium botulinum spores. Alfalfa hay that was fed to the horses contained spores and toxin.     

Outcome of equids with clinical signs of West Nile virus infection and factors associated with death

OBJECTIVE: To determine outcome of equids in the western United States with clinical signs of West Nile virus (WNV) infection and identify factors associated with risk of death in infected equids. DESIGN: Cross-sectional study. ANIMALS: 484 equids in Nebraska and Colorado. PROCEDURE: Owners of 484 equids with laboratory-confirmed West Nile virus infection in Nebraska and Colorado were contacted by telephone, and a questionnaire was used to obtain information on signalment, management, clinical signs, date of disease onset, duration of disease, WNV vaccination status, and health status at the time of the interview. RESULTS: 137 of 482 (28.4%) animals died or were euthanatized. Ataxia, lethargy, muscle fasciculations, and weakness were the most common clinical signs of disease. Animals > or = 3 years old were more likely to die than were animals < or = 2 years old. Unvaccinated equids were twice as likely to die as were animals that had been vaccinated at least once prior to the onset of disease. Animals that were recumbent and unable to rise were 78 times as likely to die as were animals that never lost the ability to rise. Females were 2.9 times as likely to die as males. Two hundred seventy-one of 339 (79.9%) animals that survived recovered fully; mean duration of disease for these animals was 22.3 days. CONCLUSIONS AND CLINICAL RELEVANCE: Among equids with WNV infection, age, vaccination status, an inability to rise, and sex were associated with the risk of death.     

Strangulating obstruction caused by intestinal herniation through the proximal aspect of the cecocolic fold in 9 horses

OBJECTIVE: To report the clinical and surgical findings and outcome for horses with strangulating obstruction caused by herniation through the proximal aspect of the cecocolic fold. STUDY DESIGN: Retrospective study. ANIMALS: Nine horses. METHODS: Medical records were reviewed for clinical signs, surgical findings and technique, and outcome. Cadaver ponies and necropsy specimens were also used to study the regional anatomy of the cecocolic fold. RESULTS: The ileum and distal jejunum were strangulated in 8 horses, whereas in 1 horse the small intestine and the left ascending colons were incarcerated in a rent in the cecocolic fold. Two horses were euthanatized at surgery, 6 horses had a small intestinal resection (mean length, 3 m; range, 1.5-6.4 m) and an end-to-side jejunocecostomy, and the entrapment was reduced without resection in the horse that had small intestine and ascending colon incarceration; cecocolic fold defects were not closed. One horse was euthanatized 36 hours after surgery because of endotoxemia. Six horses were discharged; 4 were available for long-term follow-up, of which 2 were euthanatized, and 2 were euthanatized 12 and 18 months after surgery because of colic signs. Variations in thickness of the cecocolic fold were observed in specimens obtained from necropsy of other horses and ponies. CONCLUSIONS: Reasons for this defect are unknown, although observed anatomic differences in cecocolic fold thickness may contribute to the development of defects. CLINICAL RELEVANCE: Reduction of the entrapped bowel is easiest when traction is placed on the bowel at a 90 degrees to the base of the cecum. Intestinal incarceration through rents within the proximal part of the cecocolic fold should be considered as a differential diagnosis for strangulating obstruction in horses.     

Fractures of the palmar aspect of the carpal bones in horses: 10 cases (1984-2000)

OBJECTIVE: To determine clinical and radiographic findings, treatment, and outcome of horses with fractures of the palmar aspect of the radial carpal bone, with or without concurrent fractures of the palmar surfaces of the other carpal bones. DESIGN: Retrospective study. ANIMALS: 10 horses. PROCEDURE: Medical records were reviewed to obtain information on history, signalment, clinical and radiographic findings, treatment, and outcome. Follow-up information was gathered from owners and referring veterinarians. RESULTS: 7 horses became lame after recovery from general anesthesia for treatment of an unrelated problem. The remaining 3 horses developed a fore-limb lameness after falling (1 horse) or being turned out in a pasture (2 horses). Fractures involved the palmar surface of the radial carpal bone in all 10 horses; in addition, the ulnar carpal bone was affected in 2 horses, the intermediate carpal bone in 2 horses, and the distal aspect of the radius in 4 horses. None of the 4 horses treated nonsurgically returned to work, and 3 were euthanatized because of recalcitrant lameness. In the other 6 horses, fragments were removed surgically. Two were euthanatized because of continued lameness, 1 was euthanatized for other reasons, 2 were sound enough for light work, and 1 returned to athletic work. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that fractures of the palmar aspect of the carpal bones are uncommon in horses. The prognosis appears to be poor for affected horses but may be better for horses that undergo arthroscopic removal of intra-articular fragments.     

Ataxia and paresis with equine herpesvirus type 1 infection in a herd of riding school horses

An outbreak of neurologic disease associated with serologic evidence of equine herpesvirus type 1 (EHV-1) infection occurred in a herd of 46 riding school horses. Ataxia and paresis were observed in 14 geldings and 5 barren mares. Eight affected horses had distal limb edema, 1 horse had a head tilt, and 3 others had urinary incontinence. Other clinical signs included fever, depression, and inappetance in 30 horses. Seven horses with neurologic signs were treated with acyclovir. Serum neutralizing antibody titers against EHV-1 increased 4-fold between acute and convalescent samples or exceeded 1: 256 in 19 of 44 horses, confirming recent infection. A significantly greater proportion of horses that seroconverted were mares ( P = .014). Of the 19 horses exhibiting ataxia and paresis, 17 made a complete recovery, 1 made a partial recovery, and 1 was euthanized.     

Spreading of West Nile virus infection in Croatia

West Nile virus (WNV) is an emerging zoonotic pathogen with rapid global expansion. The virus circulation is confirmed in many countries of Mediterranean Basin and Southern and Central Europe. In our study detection of specific WNV antibodies was performed in horses and cattle sera samples collected from October 2010 to April 2011. Serum samples were randomly taken from different parts of Croatia and tested by IgG and IgM ELISA. Positive serological results were confirmed by virus neutralization assay (VN-assay) and plaque reduction neutralization test (PRNT). Results showed that WNV antibodies were present in 72 out of 2098 horse sera (3.43%) and 3 of 2695 cattle sera (0.11%). The highest seroprevalence was found in Eastern Croatia in counties next to Hungarian, Serbian and Bosnia and Herzegovinian state borders. In Adriatic part of Croatia positive animals were found only in the westernmost county, near Slovenian and Italian borders. Geographic distribution and number of positive horses indicated that WNV is highly present in Croatia and spreading from East to West. However, positive horses in westernmost part of country indicate possible second origin of spreading. Location of serological positive cattle supports the hypothesis that seropositive cattle could be indicators of high WNV activity in the respective geographic regions.     

An outbreak of type C botulism in 12 horses and a mule

A USDA Early Response Team investigated deaths of several horses and a mule in northern Arizona at the request of local animal health officials. Thirteen animals (12 horses and 1 mule) housed at 5 facilities in a 7.4 square mile area died between August 1998 and January 1999. Clinical signs consisted of muscular weakness that rapidly progressed to lateral recumbency. Ten animals had paresis of the tongue, throat, or lips. Affected animals appeared alert and were interested in eating and drinking, even while recumbent. All 13 animals were euthanatized. Clostridium botulinum type C was isolated from feces or intestinal contents from 3 affected horses. Preformed toxin was detected in samples of soil and bird droppings collected from a nearby horse burial site. It was hypothesized that the outbreak was a result of birds, presumably ravens, feeding at the burial site and at horse facilities in the area that transferred toxin to the affected animals.     

West Nile virus infection in reindeer (Rangifer tarandus).

West Nile virus (WNV) infection in 4 reindeer (Rangifer tarandus) resulted in lymphohistiocytic encephalomyelitis within the medulla oblongata and cervical spinal cord. Immunohistochemistry revealed WNV antigen within neurons and among mononuclear cell infiltrates. These represent the first known cases of clinical WNV infection in Cervidae. Clinical signs and lesions were similar to those described in horses. Nucleotide sequence of a 768-bp region of the WNV E-glycoprotein gene revealed 1 nucleotide mutation, which resulted in a single amino acid substitution from a serine to a glycine (position 227 of E-glycoprotein) when compared with the prototype WNV-NY99 strain (isolated from Bronx zoo flamingo 382-99).     

Efficacy, duration, and onset of immunogenicity of a West Nile virus vaccine, live Flavivirus chimera, in horses with a clinical disease challenge model

REASON FOR PERFORMING STUDY: West Nile virus (WNF) is a Flavivirus responsible for a life-threatening neurological disease in man and horses. Development of improved vaccines against Flavivirus infections is therefore important. OBJECTIVES: To establish that a single immunogenicity dose of live Flavivirus chimera (WN-FV) vaccine protects horses from the disease and it induces a protective immune response, and to determine the duration of the protective immunity. METHODS: Clinical signs were compared between vaccinated (VACC) and control (CTRL) horses after an intrathecal WNV challenge given at 10 or 28 days, or 12 months post vaccination. RESULTS: Challenge of horses in the immunogenicity study at Day 28 post vaccination resulted in severe clinical signs of WNV infection in 10/10 control (CTRL) compared to 1/20 vaccinated (VACC) horses (P<0.01). None of the VACC horses developed viraemia and minimal histopathology was noted. Duration of immunity (DPI) was established at 12 months post vaccination. Eight of 10 CTRL exhibited severe clinical signs of infection compared to 1 of 9 VACC horses (P<0.05). There was a significant reduction in the occurrence of viraemia and histopathology lesion in VACC horses relative to CTRL horses. Horses challenged at Day 10 post vaccination experienced moderate or severe clinical signs of WNV infection in 3/3 CTRL compared to 5/6 VACC horses (P<0.05). CONCLUSIONS: This novel WN-FV chimera vaccine generates a protective immune response to WNV infection in horses that is demonstrated 10 days after a single vaccination and lasts for up to one year. POTENTIAL RELEVANCE: This is the first USDA licensed equine WNV vaccine to utilise a severe challenge model that produces the same WNV disease observed under field conditions to obtain a label claim for prevention of viraemia and aid in the prevention of WNV disease and encephalitis with a duration of immunity of 12 months.