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An unusual pattern of seroreactivity to antigens of rickettsial organisms (Rickettsia rickettsii, R rhipicephali, R montana, and R bellii), particularly to R bellii antigen, was detected in 3 dogs during a 2-month period. Thus, studies were initiated to clarify the pathogenic potential of the more distantly related rickettsial organisms (R canada and R prowazekii) in dogs. Because R bellii are nonpathogenic rickettsiae that share numerous common properties with spotted fever-group and typhus-group rickettsiae, and because closely related pathogenic relatives of R bellii have not been identified, we examined the pathogenic potential of these typhus-group rickettsiae by testing stored serum samples, by attempting rickettsial isolation from febrile dogs, and by experimentally inoculating dogs with R canada and R prowazekii.

Evaluation of results of a serosurvey of acute and convalescent serum samples from 80 dogs in which Rocky Mountain spotted fever had been considered as a differential diagnosis, but seroconversion to R rickettsii had not been documented, identified 1 dog with a fourfold increase in antibody titer to R rhipicephali and 3 dogs with fourfold increases in antibody titer to 1 or more antigens of typhus-group rickettsial organisms. A study of 15 dogs that were febrile during summer months failed to identify serologic or tissue culture evidence of typhus-group rickettsial infection or typhus-group rickettsemia, but did result in isolation of R rickettsii and Ehrlichia canis, respectively, from 1 dog each. In our final study, after experimentally inoculating 6 dogs with R canada and R prowazekii, all dogs seroconverted to the respective rickettsiae, but rickettsemia or clinical and hematologic evidence of disease was not observed.

Collectively, our results did not provide convincing evidence to support a pathogenic role for R canada and R prowazekii organisms in dogs. Our findings supported the conclusion that an unidentified microorganism, which results in production of antibodies against R bellii and antigens of typhus-group rickettsial organisms, contributed to an unexplained febrile illness of dogs in the southeastern United States. In an effort to identify dogs in which serologic evidence supports this conclusion, we recommend the use of selected spotted fever-group, typhus-group, and R bellii antigens for serodiagnostic purposes. We also recommend that diagnosticians attempt tissue culturing to isolate organisms when a rickettsial agent is suspected.

Free access
in Journal of the American Veterinary Medical Association



To compare the performance of 5 synthetic peptide–based ELISAs with that of 3 commercially available immunofluorescent assays (IFAs) for serologic diagnosis of anaplasmosis and ehrlichiosis in dogs.


A convenience set of 109 serum samples obtained before and at various times after inoculation for 23 dogs that were experimentally infected with Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Ehrlichia chaffeensis, or Ehrlichia ewingii and 1 uninfected control dog in previous studies.


All serum samples were assessed with 5 synthetic peptide–based ELISAs designed to detect antibodies against A phagocytophilum, A platys, E canis, E chaffeensis, and E ewingii and 3 whole organism–based IFAs designed to detect antibodies against A phagocytophilum, E canis, and E chaffeensis. The species-specific seroreactivity, cross-reactivity with the other tick-borne pathogens (TBPs), and diagnostic sensitivity and specificity were calculated for each assay and compared among assays.


All serum samples obtained from dogs experimentally infected with a TBP yielded positive results on a serologic assay specific for that pathogen. In general, sensitivity was comparable between ELISAs and IFAs and tended to increase with duration after inoculation. Compared with the IFAs, the corresponding ELISAs were highly specific and rarely cross-reacted with antibodies against other TBPs.


Results suggested that peptide-based ELISAs had enhanced specificity relative to whole organism–based IFAs for detection of antibodies against Anaplasma and Ehrlichia spp, which should facilitate accurate diagnosis and may help detect dogs coinfected with multiple TBPs.

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in American Journal of Veterinary Research


Objective—To determine the prevalence of stray dogs in eastern Tennessee seropositive to Ehrlichia canis and examine the correlation between results for an ELISA, indirect immunofluorescent antibody (IFA) test, and polymerase chain reaction (PCR) assay.

Sample Population—Blood samples obtained from 90 adult dogs admitted to an animal shelter in eastern Tennessee.

Procedure—Serum samples were analyzed for antibodies against E canis by use of a commercially available ELISA kit, 2 IFA tests, and a PCR assay; testing was performed at the University of Tennessee (TN) and North Carolina State University (NCSU). The PCR amplification was performed by use of DNA extracted from EDTA-anticoagulated blood and primers designed to amplify DNA of Ehrlichia spp.

Results—Antibodies against E canis were detected in only 1 dog by use of the ELISA. By IFA testing at TN, 10 of 90 (11%) dogs were seroreactive against E canis antigens, all of which had medium to high titers to E canis. Only 5 of the 10 TN seroreactors were also reactive against E canis antigens in IFA tests conducted at NCSU, and all 5 had low to medium titers. The DNA of Ehrlichia spp was not amplified in any blood samples by use of PCR assays conducted at the TN or NCSU.

Conclusions and Clinical Relevance—The discordant ELISA, IFA, and PCR results obtained in this study were unexpected and may have been related to exposure of dogs to an Ehrlichia species other than E canis, such as E ewingii. (Am J Vet Res 2004;65:1200–1203)

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in American Journal of Veterinary Research


Objective—To examine the correlation between results for an indirect immunofluorescence assay (IFA) that uses Ehrlichia canis antigen as a substrate (ie, E canis-IFA), 2 western blot (WB) analyses, and a commercially available ELISA in the detection of E canis antibody in dog sera.

Sample Population—54 canine serum samples that were reactive on E canis-IFA and 16 canine serum samples that were E canis-IFA nonreactive.

Procedure—Serum samples were evaluated by use of 2 WB analyses and a commercially available ELISA. Correlation between results of the 3 testing modalities (ie, IFA, WB analyses, and the ELISA) was examined by use of nonreactive (E canis-IFA reciprocal titer, < 20), low-titer (reciprocal titer, 80 to 160), medium-titer (reciprocal titer, 320 to 2,560), and high-titer (reciprocal titer, 5,120 to > 20,480) serum samples.

Results—For all serum samples in the nonreactive (n = 16), medium-titer (17), and high-titer (18) groups, correlation of results among IFA, WB analyses, and the commercially available ELISA was excellent. A poor correlation was found between IFA results and those of WB analyses and the ELISA for serum samples in the low-titer group (19), with only 4 of the 19 serum samples having positive results on both WB analyses and the commercially available ELISA.

Conclusions and Clinical Relevance—The discrepancy between E canis-IFA, WB analyses, and the commercially available ELISA results for the low-titer serum samples may be related to a high IFA sensitivity or, more likely, a lack of specificity associated with cross-reactivity among Ehrlichia spp.

Full access
in American Journal of Veterinary Research


Objective—To determine whether the geographic distribution of deer ticks (Ixodes scapularis) was associated with the distribution of dogs seropositive for various tick-transmitted disease organisms (ie, Borrelia burgdorferi, Rickettsia rickettsii, the human granulocytic ehrlichiosis [HGE] agent, Ehrlichia canis, and Bartonella vinsonii subsp berkhoffii).

Design—Serologic survey.

Sample Population—Serum samples from 277 dogs in animal shelters and veterinary hospitals in Rhode Island.

Results—Overall, 143 (52%) dogs were seropositive for B burgdorferi, 59 (21.3%) were seropositive for R rickettsii, 40 (14.4%) were seropositive for the HGE agent, 8 (2.9%) were seropositive for E canis, and 6 (2.2%) were seropositive for B vinsonii. Regression analysis indicated that the natural logarithm of nymphal deer tick abundance was correlated with rate of seropositivity to the HGE agent and to B burgdorferi but not to rate of seropositivity to R rickettsii, E canis, or B vinsonii. Percentages of samples seropositive for B burgdorferi, R rickettsii, the HGE agent, and E canis were significantly higher for samples from the southwestern part of the state where ticks in general and deer ticks in particular are abundant than for samples from the northern and eastern portions of the state, where ticks are relatively rare.

Conclusions and Clinical Relevance—Results suggested that all 5 disease agents are in Rhode Island and pose a risk to dogs and humans. Knowledge concerning tick distributions may be useful in predicting the pattern of disease associated with particular tick species and may aid diagnostic, prevention, and control efforts. (J Am Vet Med Assoc 2001;218:1092–1097)

Full access
in Journal of the American Veterinary Medical Association


Objective—To determine historical, physical examination, hematologic, and serologic findings in dogs with Ehrlichia ewingii infection.

Design—Retrospective study.

Animals—15 dogs.

Procedure—In all dogs, infection with E ewingii was confirmed with a polymerase chain reaction (PCR) assay. Follow-up information and clarification of information recorded in the medical records was obtained by telephone interviews and facsimile correspondence with referring veterinarians and owners.

Results—Fever and lameness were the most common findings with each occurring in 8 dogs. Five dogs had neurologic abnormalities including ataxia, paresis, proprioceptive deficits, anisocoria, intention tremor, and head tilt. Neutrophilic polyarthritis was identified in 4 dogs. No clinical signs were reported in 3 dogs. The predominant hematologic abnormality was thrombocytopenia, which was identified in all 12 dogs for which a platelet count was available. Reactive lymphocytes were seen in 5 of 13 dogs. Concurrent infection with another rickettsial organism was identified in 4 dogs. Of the 13 dogs tested, 7 were seroreactive to E canis antigens. Morulae consistent with E ewingii infection were identified in neutrophils in 8 dogs. Treatment with doxycycline, with or without prednisone, resulted in a rapid, favorable clinical response in the 9 dogs for which follow-up information was available.

Conclusions and Clinical Relevance—Results suggest that PCR testing for E ewingii infection should be considered in dogs with fever, neutrophilic polyarthritis, unexplained ataxia or paresis, thrombocytopenia, or unexplained reactive lymphocytes, and in dogs with clinical signs suggestive of ehrlichiosis that are seronegative for E canis. Following treatment with doxycycline, the prognosis for recovery is good. (J Am Vet Med Assoc 2003;222:1102–1107)

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in Journal of the American Veterinary Medical Association



To characterize the pathogenic potential of a unique Borrelia isolate obtained from a dog from Florida (FCB isolate).


Prospective experimental infection.


32 preweanling Swiss Webster mice and 12 adult male Hartley guinea pigs were injected intraperitoneally with 105 spirochetes.


Mice were used as controls and blood recipients, and at 3- to 4-day intervals, 1 control mouse and 2 infected mice were necropsied, tissues were cultured, and a recipient mouse was inoculated with blood. Guinea pigs were randomized to 4 groups and inoculated intradermally with 100, 102, 103, or 104 spirochetes. For 48 days, clinical, hematologic, serologic, and microbiologic tests were performed on them, after which they were necropsied.


In mice, spirochetemia was detectable between postinoculation days (PID) 3 and 13, and seroreactivity to homologous antigen was detectable during PID 10 through 31. Compared with control mice, infected mouse spleens were 2 to 3 times larger. Histologic lesions included lymphoid hyperplasia, neutrophilic panniculitis, epicarditis, and myocarditis, with intralesional spirochetes detected from PID 3 through 6. During PID 10 through 31, nonsuppurative epicarditis developed. Signs of illness and hematologic abnormalities were not observed in guinea pigs, despite isolating spirochetes from blood during PID 7 to 27. When necropsied on PID 48, histologic lesions included lymphoid hyperplasia and lymphocytic plasmacytic epicarditis.


The FCB isolate causes spirochetemia, lymphoid hyperplasia, dermatitis, and myocardial injury in Swiss Webster mice and can be transmitted by blood inoculation. In Hartley guinea pigs, the isolate causes spirochetemia, lymphoid hyperplasia, and epicarditis. Documentation of disease in mice, guinea pigs, and, presumably, dogs raises the level of concern that the FCB isolate might be pathogenic for man and other animal species. (Am J Vet Res 1996;57:505–511)

Free access
in American Journal of Veterinary Research