Serosurvey of horses with evidence of equine monocytic ehrlichiosis

In August 1986, an extensive serosurvey for prevalence of IgG and IgM antibodies against Ehrlichia risticii, the causative agent of equine monocytic ehrlichiosis (EME), was performed at 2 Ohio racetracks, River Downs (RD) and Beulah Park (BP). Of 840 horses at RD and 574 at BP, 13 and 20%, respectively, were IgG antibody-positive (by indirect fluorescent antibody test results), with antibody titer ranging from 1:20 to 1:10,240. The titer observed at highest frequency at both racetracks was 1:80. A higher proportion of horses was ill at RD (operating during the summer months) than at BP (winter track). Of ill horses, 41% (24/58) at RD and 58% (11/19) at BP were seropositive. At RD, 70% (589/840) of all horses and 95% (102/107) of IgG seropositive horses had been stabled only at RD during the month prior to testing. Analysis of these sera by use of an ELISA to detect IgM antibody against E risticii antigen indicated that at RD, 42% (57/137) of the seropositive horses were IgM seropositive. At BP, 17% (20/120) of seropositive horses were IgM seropositive. The larger number of IgM seropositive horses at RD indicates that more horses were recently infected at RD than at BP (P = 0.0001). Therefore, at least half the seropositive horses at RD seemed to have acquired the infection at RD. These serosurvey data also indicate that at BP and RD, 78% (85/109) and 91% (111/122) of IgG seropositive horses, respectively, had subclinical infection. At less than or equal to 1:40 titer, there was no difference in seropositive rates between healthy and ill horses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Disease features in horses with induced equine monocytic ehrlichiosis (Potomac horse fever)

Fifty-five horses were inoculated IV and/or SC with materials containing Ehrlichia risticii, ie, infected whole blood, buffy coat cells, or cell culture, to study clinical and hematologic features of equine monocytic ehrlichiosis (Potomac horse fever). Major clinical and hematologic features of induced E risticii infection were biphasic increase in rectal temperature with peak increases of 38.9 C and 39.3 C on postinoculation days (PID) 5 and 12, respectively; depression; anorexia; decreased WBC count (maximal decrease of 47% on PID 12); and diarrhea from PID 14 to PID 18. Increased WBC count was an inconsistent feature, with a maximal increase of 51.5% on PID 20. During times of decreased and increased WBC counts, lymphocyte/neutrophil ratios remained fairly constant. However, not all horses had all clinical and hematologic features, and these features were present in different degrees among horses. Increased rectal temperature, depression, anorexia, and decreased WBC count were more consistent features, whereas diarrhea developed in 73% of the horses. Of 55 horses, 39 (71%) had all clinical and hematologic features of the disease (classic disease), whereas 16 (29%) horses did not have greater than or equal to 1 of these features (nonclassic disease). The E risticii titer in the blood (ehrlichemia) was maximum during the peak increase in rectal temperature. In 55 horses, mortality was 9%. Significant differences (P greater than 0.5) in clinical and hematologic features were not detected between horses that survived and those that died of E risticii infection.     

Risk factors for equine monocytic ehrlichiosis seropositivity in horses in Colorado

A study was conducted in April through August 1989 whereby 318 horses were tested for immunofluorescent antibody (IFA) titers to Ehrlichia risticii. The state of Colorado was divided into four regions and an approximately equal number of horses was tested in each region. Questionnaires were completed for each horse; a multiple logistic regression model was used to identify risk factors for horses being exposed to E. risticii.

Of the 318 horses tested, 48 (16%) had IFA titers of 1:40 or greater (range 1:40–1:1280). There was no association between the region of Colorado and seropositivity. Other variables which were not identified as risk factors included gender, transportation to other parts of Colorado, transportation to other states and exposure to other horses. When the variables were examined individually, older horses, Quarter horses, horses housed at a boarding stable, and horses having access to pasture or direct access to a stream, pond or lake were more likely to have positive titers. Horses confined to a stall with a run were less likely to be exposed to E. risticii. All apparent associations disappeared when the factors were tested in the multiple logistic regression model.     

Serodiagnosis of equine monocytic ehrlichiosis in selected groups of horses in Minnesota

Antibody titer to Ehrlichia risticii was determined, in 2,549 equine serum samples, using an indirect fluorescent antibody assay. During 1986, samples were obtained from the Minnesota State-Federal Equine Infectious Anemia Diagnostic Laboratory, the Minnesota Racing Laboratory, from horses admitted to the University of Minnesota Veterinary Teaching Hospital, and as a result of field investigations of horses with acute diarrhea. Results of the study revealed antibody prevalence of 33, 24, 47, and 25% for the respective groups. There was no statistical association between seropositive status and age, sex, breed, or clinical problem of horses referred to the teaching hospital. There was an increase in the total percentage of seropositive samples over the duration of the sample collection period, suggesting a seasonal exposure pattern, and E risticii was associated with clinical and subclinical infections in horses of Minnesota.     

Diagnosis of equine monocytic ehrlichiosis (Potomac horse fever) by indirect immunofluorescence

The recent establishment of a system for the continuous in vitro propagation of Ehrlichia risticii, the causative agent of equine monocytic ehrlichiosis (EME; synonym, Potomac horse fever), has facilitated the development of an indirect fluorescent antibody test for the diagnosis of this disease under laboratory and field conditions. The field diagnostic application of the test has aided in the recognition of the disease in 16 states of the United States and in 1 province of Canada. A limited epidemiologic study conducted between January and September 1985, in an area where the disease is known to be enzootic, revealed that conversion from seronegative to seropositive status is not always accompanied by clinical manifestations of the disease. Confirmatory findings in experimentally inoculated horses suggest the existence of clinically undetectable infections.     

IgG antibody subclass response against equine herpesvirus type 4 in horses

In this study, IgG subclass responses against equine herpesvirus type 4 (EHV-4) were examined by enzyme-linked immunosorbent assay (ELISA) using a type-specific region of EHV-4 glycoprotein G (gG). ELISA using sera collected from horses experimentally infected with EHV-4 revealed that IgGa and IgGb antibodies were detected at high level, but IgGc and IgG(T) antibody responses were detected at low level or were undetectable. The IgGa antibody response reached its peak on day 10 post-infection, and then dropped. The IgGb antibody response reached its maximum level on day 12 post-infection, and then the level was sustained during at least 28 days after infection. Forty healthy racehorses that had already been infected with EHV-4 possessed antibody against EHV-4. Although IgGa antibodies specific for EHV-4 were not detected in any horses, IgGb antibodies were detected and the levels correlated with total IgG antibodies against EHV-4 gG. The results suggest that EHV-4-specific IgGa and IgGb antibodies are induced in EHV-4-infected horses, and that IgGb antibody, but not IgGa, is long lasting.     

The effect of age on serum antibody titers after rabies and influenza vaccination in healthy horses

BACKGROUND: The proportion of geriatric horses within the equine population has increased in the past decade, but there is limited information on the immune function of these animals. HYPOTHESIS: Aged horses will have a lesser increase in serum antibody response to vaccination. ANIMALS: Thirty-four aged healthy horses (> or = 20 years) and 29 younger adult horses (4-12 years) of various breeds. METHODS: All horses were vaccinated with vaccines of killed rabies and influenza virus. Horses in each age group were allocated to receive either rabies or influenza booster vaccine 4 weeks after the initial vaccination. Serum samples were taken at 0, 4, 8, and 24 weeks. Rabies serum neutralization titers and equine influenza virus specific antibody sub-isotypes (IgGa, IgGb, IgG(T), and IgA) as well as single radial hemolysis (SRH) titers were determined. RESULTS: Rabies antibody titers were similar in the 2 age groups at all sampling times. Aged horses had higher IgGa and IgGb influenza antibody titers before vaccination than younger horses but similar titers after vaccination (P= .004 and P= .0027, respectively). Younger horses had significantly greater increases in titer than aged horses at all sampling times for IgGa (P= .001) and at 8 and 24 weeks for IgGb (P= .041 and .01, respectively). There was no detectable serum IgG(T) at any time point. A significant booster vaccine effect was seen for both antirabies and anti-influenza titers. Anti-influenza titer before vaccination also had a significant effect on subsequent antibody response. CONCLUSIONS AND CLINICAL IMPORTANCE: Healthy aged horses generated a primary immune response to a killed rabies vaccine similar to that of younger adult horses. Aged horses had a significantly reduced anamnestic response to influenza vaccine.     

Susceptibility of dogs to infection with Ehrlichia risticii, causative agent of equine monocytic ehrlichiosis (Potomac horse fever)

Adult dogs 1 to 5 were inoculated IV and/or SC with 3, 5, or 6 ml of a suspension containing 1.2 x 10(4) Ehrlichia risticii-infected cells (derived from primary canine monocyte cell cultures)/ml. Dogs 6 to 8 were inoculated IV and/or SC with 3 or 6 ml of 1.2 x 10(5) organism-free cultured canine monocytes/ml. Ehrlichia risticii was isolated in cultures from inoculated dogs 3, 4, and 5 on postinoculation days (PID) 10 to 16, but not from dogs 6 to 8. Dogs inoculated with E risticii seroconverted between PID 6 and 12. Clinical signs of illness were not observed in these 5 E risticii-inoculated dogs. A pony, inoculated with E risticii isolated from inoculated dog 5, developed clinical signs of equine monocytic ehrlichiosis, including fever, anorexia, depression, and diarrhea, and E risticii was isolated from the pony's blood. This E risticii isolate was then inoculated into susceptible dog 9, and E risticii was repeatedly isolated from dog 9 during PID 6 to 17. Dogs were susceptible to infection with E risticii and may serve as a reservoir of the organism in the field.     

High prevalence of bovine papillomaviral DNA in the normal skin of equine sarcoid-affected and healthy horses

Bovine papillomavirus (BPV), the causative agent of papillomas in cattle, has been shown to play a major role in the pathogenesis of equine sarcoids in horses. BPV has also been detected occasionally in normal equine skin. In this study, presence and activity of BPV in normal skin and peripheral blood of 4 groups of horses were evaluated: sarcoid-affected horses, horses living in contact with sarcoid-affected horses, horses living in contact with papilloma-affected cattle and control horses. From each horse, 3 samples on 4 locations were collected: a swab of the intact skin surface and both a swab and a biopsy after decontamination. BPV DNA was found in the normal skin of 24 of 42 horses (57%). Mainly sarcoid-affected horses and horses living in contact with cattle were carriers (73%), but BPV DNA was also detected in 50% of the horses living in contact with sarcoid-affected horses and in 30% of the control population. BPV mRNA was detected in 58% of the samples positive for BPV DNA, although in a much lower quantity compared to sarcoids. In most of the BPV DNA positive samples mild acanthosis, slight basophilic cytoplasmic swelling of the epidermal layers and/or thickening of the basal membrane were noticed, but these observations were also present in several BPV DNA negative normal skin samples. BPV DNA could not be detected in peripheral blood. These findings suggest latent infection and a wide-spread occurrence of BPV in the horse population.     

The prevalence and anatomical distribution of equine gastric ulceration syndrome (EGUS) in 201 horses in Denmark

Reasons for performing study: The prevalence (up to 93% in Thoroughbred racehorses) and severity of equine gastric ulceration syndrome (EGUS) have been correlated with the type of training and associated management practices. However, there have been few reports to confirm these findings in nonracehorses in Europe. Objectives: To describe the prevalence, anatomical distribution, severity and number of gastric ulceration lesions in a population of Danish pleasure horses; and to investigate differences for groups based on age, breed type and workload. Methods: A total of 201 horses not in active race‐training, age 7 months‐27 years, were evaluated, representing 23 different stables from all 5 regions of Denmark. These horses were considered to be healthy by the owner and not on veterinary treatment for EGUS. Endoscopically observed ulcer lesion scores were based on the number present (0–4) and severity (0–5). The presence or absence of ulcers in the glandular and/or nonglandular regions of the stomach was recorded and which site the most severe ulcers were found. Results: The prevalence of EGUS severity score ≥2 was 53%. The most severe lesions were commonly observed at the margo plicatus. Although older horses were not more likely to be affected by clinically significant EGUS they were more likely to have lesions in both the glandular and nonglandular regions. Differences in location of EGUS lesions were identified in different age groups, breed types and in horses exposed to different levels of work. Conclusion and potential relevance: This study confirms that gastric ulceration can be prevalent in a group of apparently clinically normal horses, not in intensive work. Further investigation of reasons for differences in EGUS location between different populations may aid toward the development of novel preventive measures.     

The classification of seven serotypes of equine encephalosis virus and the prevalence of homologous antibody in horses in South Africa

Selected isolates of equine encephalosis virus were shown to have comparable viral protein profiles and to represent seven distinct serotypes, based on cross-neutralization tests. Serotype-specific virus-neutralizing antibody in serum samples from horses confirmed the widespread occurrence of infection. The distribution and prevalence of individual serotypes however, varied considerably. Localised foci with an increased seasonal seroconversion in groups of horses to a specific serotype and the detection of an ongoing low level of infection from other serotypes within the population, confirmed the independent persistence of the viruses in a maintenance cycle. The identification of donors with antibody resulting from infection with multiple serotypes indicated a low level of cross-protection in horses to natural reinfection.     

The prevalence of methicillin-resistant staphylococci in healthy horses in the Netherlands

Two hundred healthy horses housed at 23 different farms and one clinic and 42 persons in close contact with these horses were screened for the presence of methicillin resistant staphylococci. Samples were taken from the nose and the pastern of the horses and from the nose and throat of the humans and incubated in selective media. Isolates were identified by standard techniques and their susceptibilities were tested using an agar diffusion method. Methicillin-resistant strains were tested for the presence of the mecA gene by PCR. In 45 horses (22.5%) and 15 humans (35.7%) mecA positive staphylococci were found. All isolates were coagulase negative staphylococci, except for one methicillin-resistant Staphylococcus aureus isolated from a veterinarian. Staphylococcus sciuri was the predominant species found among the methicillin resistant staphylococci (MRS) in the horses, whereas S. epidermidis predominated in the humans. From the horses, often more than one species of MRS could be isolated, resulting in a total of 175 mecA positive equine isolates. The equine isolates were predominantly susceptible to most antimicrobials tested, whereas the human isolates showed more resistance. In conclusion, no methicillin-resistant Staphylococcus aureus was found in healthy horses in the Netherlands, but methicillin-resistant coagulase negative staphylococci were found frequently. Further studies are needed in order to investigate whether horses can be a reservoir for MRS or the mecA gene for humans.     

Pericarditis in horses: six cases (1982-1986)

Records of 6 horses with pericarditis were reviewed. Septic pericarditis was suspected in all horses, based on historic and clinical findings. In horses 1, 2, and 4, cytologic examination of the pericardial effusion revealed acute inflammation with severe neutrophil degeneration. In horses 3 and 5, cytologic examination of pericardial fluid revealed subacute inflammation with degenerated neutrophils, and in horse 6, chronic active inflammation, with well preserved neutrophils. In horses 1 and 3, bacteria were identified on cytologic examination of pericardial fluid. Results of microbiologic cultures of pericardial fluid were positive in horse 3. All horses were treated with broad-spectrum antibiotics. An indwelling pericardial catheter was used to lavage and directly administer antibiotics into the pericardial sac. Horses 1, 4, 5, and 6 survived, horse 2 died of unrelated causes, and horse 3 was euthanatized at the owner's request. Surviving horses returned to athletic performance.     

Cholelithiasis in horses: ten cases (1982-1986)

Ten horses with clinical signs consistent with cholelithiasis were evaluated. Fever, icterus, mild intermittent colic, and weight loss were reported. Clinical laboratory abnormalities included leukocytosis, hyper-proteinemia, and hyperfibrinogenemia. Gamma glutamyltransferase and liver isoenzyme of lactate dehydrogenase activities also were high. Choleliths were observed via ultrasonography of the liver in 5 of the 8 horses evaluated, and increased echogenicity of the hepatic parenchyma and dilated bile ducts were observed in all horses. Seven horses were treated medically, 5 of which died or were euthanatized. Three horses were treated surgically, of which only 1 survived. Cholelith composition varied, but cholesterol, calcium bilirubinate, and mixed bile pigments were most commonly observed.     

Bone marrow mastocytosis in dogs with myelosuppressive monocytic ehrlichiosis (Ehrlichia canis): a retrospective study

BACKGROUND: Bone marrow mastocytosis has been reported rarely in naturally occurring canine monocytic ehrlichiosis (CME). OBJECTIVES: The aims of the present study were to estimate the prevalence and magnitude of bone marrow mastocytosis in a case series of dogs with natural CME and to assess the association, if any, between mastocytosis and the clinical severity of the disease. METHODS: Seventy-six dogs with confirmed CME (Ehrlichia canis) were included in the study. Affected dogs were allocated into group A (n = 51) without bone marrow hypoplasia and group B (n = 25) with bone marrow hypoplasia. Twenty clinically healthy Beagles not previously exposed to E canis served as controls (group C). The main inclusion criteria for group A were documentation of normocellular to hypercellular bone marrow and complete clinical cure following a 4-week treatment with doxycycline, while those for group B were bone marrow hypoplasia and lack of response to doxycycline. Bone marrow aspirate smears from all 96 dogs were Giemsa-stained and examined for the presence of mast cells, which were calculated as a percentage of 1,000 nucleated cells (NCs). The prevalence of mastocytosis was compared among the 3 groups by the Pearson's chi-square test. RESULTS: Bone marrow mastocytosis (>0.1% of NCs) was found in 5 (20%) dogs in group B (range, 0.5-2.5% of NCs; median, 1% of NCs). One dog in each of groups A and C had 0.1% mast cells in the marrow. The prevalence of bone marrow mastocytosis in dogs in group B was significantly higher (P = .004) than in groups A and C. CONCLUSION: Bone marrow mastocytosis can be seen in a substantial number of dogs with E canis-induced myelosuppression.