Sixteen healthy ponies were inoculated IV with Ehrlichia risticii-infected P388D1 mouse monocytes. Of the 16 ponies, 15 developed clinical signs of equine ehrlichial colitis. Twenty-four hours after onset of fever (rectal temperature > 38.8 degrees C), 7 ponies were treated with 25 mg of erythromycin stearate/kg of body weight and 10 mg of rifampin/kg, given orally every 12 hours for 5 days. The remaining 8 ill ponies served as nontreated controls. All ponies were observed for progression of clinical signs typical of equine ehrlichial colitis. Within 12 hours of initiation of treatment, 4 of the 7 treated ponies had rectal temperature < 38.4 C and, within 24 hours, 6 of the 7 ponies had rectal temperature < 38.3C. In contrast, all control ponies had rectal temperature > 39.2 C at 24 hours (P < 0.05). Of the 7 treated ponies, 4 no longer had signs of mental depression after the second day of treatment, and only 1 of the 7 ponies had mild signs of depression after the third day of treatment. In contrast, control ponies had high mental depression score during the observation period (P < 0.05). Feed intake improved in ponies of the treatment group, with feed intake of 4 of the 7 ponies returning to normal; the other 3 ponies were only mildly anorectic by the second day of treatment. Control ponies progressively decreased their feed intake during the observation period (P < 0.05). One control pony and 2 treated ponies developed diarrhea before the treatment/observation period began. Only 1 treated pony developed diarrhea after treatment began. Of the 8 control ponies, 7 developed diarrhea. Profound decrease in borborygmal sounds with silent periods lasting longer than 3 minutes was observed in 7 of the 8 control ponies. Only 1 of the 7 treated ponies had such profound decrease in borborygmi (P < 0.05). The decrease in borborygmal sounds progressed in the control ponies during the observation period. None of the treated ponies continued to have decreased borborygmi after treatment day 2 (P < 0.05). Of the 8 control ponies, 2 were euthanatized; all treated ponies survived. In survivors, signs lasted 8 to 17 (mean, 10) days in control ponies but only 1 to 5 (mean, 2.9) days in treated ponies.

Fetal infectivity of Ehrlichia risticii was investigated in 19 ponies that were E risticii negative on the basis of results of an indirect fluorescent antibody (IFA) test. Thirteen pregnant ponies were infected by IV administration of E risticii between 90 and 180 days of gestation. Six pregnant ponies served as noninfected controls. Each infected pony had clinical signs of equine monocytic ehrlichiosis, was confirmed to be ehrlichemic, and developed an IFA titer to E risticii. Two infected ponies became recumbent, were unresponsive to supportive care, and were euthanatized. After recovery from clinical illness, the remaining ponies were observed throughout gestation for reproductive abnormalities. On abortion, each fetus was necropsied and tissue specimens from the liver, bone marrow, spleen, colon, and mesenteric lymph nodes were inoculated into canine monocyte cell cultures. Six infected ponies aborted at a mean 217 days of gestation, which was between postinoculation days 65 and 111. Five fetuses were recovered for evaluation, and E risticii was isolated from 4 of them. All 5 fetuses recovered had similar histologic findings, including enterocolitis, periportal hepatitis, and lymphoid hyperplasia with necrosis of the mesenteric lymph nodes and spleen. All fetuses tested negative for IgG to E risticii, although 3 had low IgM titer to E risticii. The remaining 5 infected ponies had normal parturition. Presuckle IFA titer to E risticii was measured in 4 of the term foals, and results for 3 were positive. Two foals from infected ponies were monitored for 6 months and daily gain in body weight was comparable to that of a control foal. None of the control ponies became ill or seroconverted during the clinical illness phase, and none aborted throughout gestation. Two control ponies seroconverted to E risticii 6 weeks before parturition. Results of this study indicate that E risticii is a primary abortifacient under experimental conditions.

Eighteen ponies were inoculated IV with Ehrlichia risticii-infected P388D1 mouse monocyte cells. Twenty-four hours after onset of fever (rectal temperature > 38.8 C), 9 ponies were treated with oxytetracycline (6.6 mg/kg of body weight, IV, q 24 h) for 5 days. The remaining 9 ponies served as infected nontreated controls. Mean scores of the following variables were not significantly different between groups on the day treatment was begun: rectal temperature, diarrhea, borborygmal sounds, feed intake, mental attitude, and evidence of a hyperresonant area in the abdomen. All ponies were observed for progression of clinical signs typical of ehrlichial colitis. Within 12 hours of initiation of treatment, only 1 treated pony had a rectal temperature > 38.8 C and most rectal temperatures were < 38.3 C. In contrast, only 2 control ponies had rectal temperatures < 38.8 C (mean rectal temperature values were significantly, P = 0.01, different between groups). In the treatment group, 4 ponies had no signs of depression after the first day of treatment, and only 1 had signs of depression after the second day of treatment (mean scores between groups were significantly different, P = 0.01). Feed intake remained normal in 4 treated ponies and improved in 4 of the remaining 5 after treatment began. Most of the control ponies had a progressive decrease in their feed intake during the observation period (mean scores between groups were significantly, P = 0.01, different). Three ponies in the control group and 2 ponies in the treatment group developed diarrhea before the treatment observation period began. Of the remaining 6 control ponies, 4 developed diarrhea after the treatment observation period began. None of the ponies in the treatment group developed diarrhea after treatment began. A profound decrease in borborygmal sounds, with silent periods lasting longer than 3 minutes, developed in 7 control ponies. Only 2 treatment ponies had borborygmi decreased to this level (mean scores between groups were significantly, P = 0.01, different). Three of 9 control group ponies developed severe disease and were euthanatized. All treatment-group ponies survived. In surviving ponies, clinical signs lasted 8 to 16 days (mean, 11.5) in the control group, but lasted only 1 to 7 days (mean, 4.5) in the treatment group (P = 0.01). Ponies from the treatment group did not develop clinical signs when reinoculated with Ehrlichia risticii at 4 and 8 months after original inoculation.

Fourteen ponies and 3 horses were inoculated with Ehrlichia risticii 2 to 20 months after a similar initial inoculation. Although all 17 had clinical signs of equine ehrlichial colitis after the first inoculation, 16 of 17 remained clinically normal following the second inoculation. The remaining pony had a transient fever and developed signs of depression. Before the initial inoculation, none of the animals had a detectable antibody titer to E risticii. All animals developed titers after the initial infection; however, a significant change of titer did not develop after reinoculation in most animals.

The seroprevalence and seasonal trend of antibody titers against equine monocytic ehrlichiosis (Potomac horse fever) were determined in apparently healthy horses in selected areas of Illinois in 1986. Sera from 1,367 horses (6 months to 29 years old) were evaluated for the presence of antibodies against Ehrlichia risticii with indirect immunofluorescence. The majority (88%) of the horses were Thoroughbred or Standardbred racehorses. The number of horses with antibodies against E risticii was 229/1,367 (16.75%). The titers in these horses ranged from 1:10 to 1:640. As the year progressed, the number of seropositive horses (titers greater than or equal to 1:10) and the magnitude of the titers increased significantly, both reaching a maximum in July and August, respectively (P less than 0.05). A relationship between seropositivity and gender was not detected. In the year prior to sampling, 56.8% of the seropositive horses had not been ill, whereas 0.8% had diarrhea, an episode of acute abdominal pain, or laminitis. It was concluded that a large number of horses in Illinois are exposed to E risticii, that maximal exposure occurs in July, and that the most common form of the disease in Illinois is not associated with clinical signs.

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 > 0.5) in clinical and hematologic features were not detected between horses that survived and those that died of E risticii infection.

A retrospective study was designed to determine the distribution of equine monocytic ehrlichiosis among the equine population in New York state, and to identify factors associated with risk of disease. Serum samples submitted to the diagnostic laboratory of the university during the period from January 1985 through December 1986 were examined for antibodies to Ehrlichia risticii, using the indirect fluorescent antibody technique. Factors evaluated included geographic origin and date of submission of the sample, and age, breed, and sex of the horse. Logistic regression analysis was used to identify which factors were significantly associated with the risk of seropositivity to E risticii, while simultaneously controlling for other factors. Of the 2,579 tested samples, 1,950 (76%) had positive results. Factors significantly associated with risk of seropositivity to E risticii were: breed of the horse (Thoroughbreds were 3 times more likely to have been exposed to E risticii, compared with non-Standardbred, non-thoroughbred breeds); sex (female horses were 2.7 times more likely to have been exposed, compared with male horses); age of the horse (the risk of being exposed to E risticii increased with age, peaked at around 12 years, and decreased thereafter); and month of submission (horses tested during November and December had the highest odds of being seropositive [odds ratio = 2.1], and horses tested during March through April were least likely to be seropositive [odds ratio = 0.5], compared with horses tested during January and February).

The antigenic profile of Ehrlichia canis, E risticii, E sennetsu, and E equi was investigated by the use of protein (western) immunoblot technique. Results of analysis of serum from acutely and chronically infected animals indicated that the 4 Ehrlichia species share a unique 25-kD polypeptide in addition to other peptides. Immune sera from dogs inoculated with E canis recognized a wide range of E canis polypeptide antigens, as determined by western blot analysis. A larger number of E sennetsu polypeptides were detected when homologous antiserum and antiserum to E equi were used. The latter antiserum did not recognize antigens of E canis or E risticii. Antisera to E canis, E risticii, and E sennetsu detected E equi antigens. Data indicate that a 25-kD protein is a common antigen among the species of the genus Ehrlichia and that the ascending order of abundance of immunodominant determinants in the 4 species of Ehrlichia studied would be: E risticii leads to E equi leads to E sennetsu leads to E canis. Implications of these findings for diagnosis of ehrlichial infections and prophylaxis are evident.