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  • Author or Editor: Jacob W. IJdo x
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Abstract

Objective—To determine whether cats in the northeastern United States develop serum antibodies against antigens of Borrelia burgdorferi and Anaplasma phagocytophilum and whether coinfection with the 2 organisms occurs.

Sample Population—Serum samples from 84 healthy cats and 9 cats with lameness, fever, anorexia, or fatigue.

Procedure—Serum antibodies against B burgdorferi and A phagocytophilum were measured with an ELISA incorporating a whole-cell preparation or purified recombinant antigens, by means of western blot analysis, or indirect fluorescent antibody (IFA) staining.

Results—ELISA results indicated that 44 of 93 (47%) sera contained antibodies against ≥ 3 B burgdorferi antigens, whereas 43 (46%) were reactive to wholecell B burgdorferi. Serum reactivity to protein 35, VlsE, and outer surface proteins A and F was most common. Seropositivity to ≥ 3 antigens occurred at the same rate (5/9) in the 9 ill cats as in the 84 healthy cats (46% [39/84]). Of 13 sera reactive to recombinant antigens, 9 were seropositive as measured by western blot testing with whole-cell antigen. Seropositivity rates of 30% and 38% were detected for antibodies against A phagocytophilum via IFA and ELISA testing, respectively. Fifteen (16%) sera had antibodies against both pathogens.

Conclusions and Clinical Relevance—Cats living in areas infested by Ixodes scapularis ticks are exposed to B burgdorferi and A phagocytophilumand, in some instances, may be coinfected. Most cats appeared healthy. An ELISA incorporating specific recombinant antigens may be used adjunctively with western blot and other assays to confirm B burgdorferi and A phagocytophilum infection in cats. (Am J Vet Res 2005;66:1895–1899)

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in American Journal of Veterinary Research

Abstract

Objective—To develop and evaluate a polyvalent ELISA incorporating a highly specific recombinant antigen (p44) for diagnosis of granulocytic ehrlichiosis in dogs and horses.

Animals—32 dogs and 43 horses.

Procedure—Results of the ELISA were compared with results of indirect fluorescent antibody (IFA) staining and western immunoblotting incorporating whole-cell antigen.

Results—For the canine and equine samples, percentages of samples with positive IFA staining, western immunoblotting, and ELISA results were similar. For 29 (91%) canine samples and 30 (70%) equine samples, results of IFA staining, western immunoblotting, and the ELISA were in complete agreement. Results of the ELISA for 3 canine serum samples known to contain antibodies to Ehrlichia canis and 12 equine serum samples known to contain antibodies to E risticii were negative.

Conclusions and Clinical Relevance—Results of the present study suggest that a polyvalent ELISA incorporating a recombinant p44 antigen is suitable for detecting antibodies to E equi in dogs and horses. ( Am J Vet Res 2001;62:29–32)

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in American Journal of Veterinary Research

Abstract

Objective—To test serum samples of dogs and horses by use of class-specific recombinant-based ELISA for establishing a diagnosis of granulocytic ehrlichiosis attributable to infection with organisms from the Ehrlichia phagocytophila genogroup.

Sample Population—Serum samples from 43 clientowned dogs and 131 horses (81 with signs of acute illness and 50 without signs of disease).

Procedure—Serum samples were analyzed, using ELISA with a recombinant 44-kd protein antigen for IgM and IgG antibodies to the human granulocytic ehrlichiosis (HGE) agent (NCH-1 strain). Western blot analyses, using infected human promyelocytic leukemia cells, were conducted on 38 serum samples of horses and 11 serum samples of dogs to verify reactivity to the 44-kd peptide.

Results—IgM or IgG antibodies to the HGE agent were detected in 5 to 28% of dog serum samples and 5 to 37% of horse serum samples. Thirty-five of 38 (92%) horse serum samples had corresponding results on both tests (2 positive results for 26 samples and 2 negative results for 9 samples), using an ELISA for IgG antibodies or immunoblotting for total immunoglobulins. All 11 serum samples of dogs had positive results for both methods.

Conclusion and Clinical Relevance—These ELISA with recombinant 44-kd antigen are suitable for detecting IgM or IgG antibodies to the HGE agent in serum samples of dogs and horses. Positive results for serum samples of horses from Connecticut, New York, Virginia, and Georgia indicate that the HGE agent is widely distributed in tick-infested areas of the eastern United States. (Am J Vet Res 2001;62:1365–1369)

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in American Journal of Veterinary Research

Abstract

Objective—To determine whether horses living in tick-infested areas of northeastern United States with clinical signs of borreliosis or granulocytic ehrlichiosis had detectable serum antibodies to both Borrelia burgdorferi and Ehrlichia equi.

Design—Prospective study.

Animals—Serum samples from 51 clinically normal horses, 14 horses with clinical signs of borreliosis, and 17 horses with clinical signs of granulocytic ehrlichiosis.

Procedure—Serum B burgdorferi or E equi antibodies were measured by use of an ELISA, immunoblot analysis, or indirect fluorescent antibody (IFA) staining.

Results—Of the 82 serum samples tested, 37 (45.1%) and 13 (15.9%) had detectable antibodies to B burgdorferior E equi, respectively. Test results indicated that 12 horses had been exposed to both agents, 11 of these horses had granulocytic ehrlichiosis. The ELISA regularly detected antibodies to the following recombinant protein (p) antigens of B burgdorferi: p29, p37, p39, and p41-G. The use of immunoblot analysis confirmed ELISA results by indicating antibody reactivities to antigens of whole-cell B burgdorferi having molecular masses of predominantly 31, 34, 37, 39, 41, 58, and 93 kd.

Conclusion and Clinical Relevance—Horses living in areas where ticks (Ixodes scapularis) abound are sometimes exposed to multiple pathogens. Analyses for specific recombinant borrelial antibodies using an ELISA can help separate horses with borreliosis from those with granulocytic ehrlichiosis, even when antibodies to both etiologic agents are detected in serum samples. Analysis using immunoblots is sensitive, and along with ELISA or IFA procedures, is suitable for confirming a clinical diagnosis of each disease (J Am Vet Med Assoc 2000;217:1045–1050)

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in Journal of the American Veterinary Medical Association

Abstract

Objective

To diagnose granulocytic ehrlichiosis in horses, compare results of indirect fluorescent antibody (IFA) staining procedures with those of immunoblot analysis, and compare serologic test findings with polymerase chain reaction (PCR) results.

Animals

69 horses with high rectal temperatures (≥ 39 C) and lethargy, anorexia, or limb edema.

Procedure

43 convalescent serum samples obtained from 38 horses 2 to 18 weeks after onset of illness were analyzed by use of immunoblot procedures and IFA staining methods, using the NCH-1 or BDS ehrlichial strains. Blood samples from 69 acutely ill horses were tested by PCR to detect ehrlichial DNA.

Results

Antibodies to Erlichial equi were detected in serum samples obtained during all seasons; seropositivity rates ranged from 50 to 93%. In IFA assays using the BDS or NCH-1 strain, seropositivity rates were 70 and 79%, respectively, whereas in immunoblot analyses using the NCH-1 strain, a seropositivity value of 79% was recorded. By immunoblot analysis, all serum samples of all seropositive horses were reactive to a protein having molecular mass of about 44 kd. Blood samples from 29 of 69 (42%) acutely ill horses contained ehrlichial DNA.

Conclusion and Clinical Relevance

Results of the various serologic testing procedures were in close agreement with each other. All serologic testing methods are suitable for laboratory diagnosis of equine granulocytic ehrlichiosis. (Am J Vet Res 1999; 60:631–635)

Free access
in American Journal of Veterinary Research

Objective

To determine whether dogs living in tick-infested areas of the northeastern United States had been exposed to Ehrlichia equi, an etiologic agent of granulocytic ehrlichiosis.

Design

Analyses of dog sera.

Animals

106 ill dogs and 12 clinically normal dogs.

Procedure

Antibodies to E equi were detected by indirect fluorescent antibody staining methods and western blot analyses.

Results

10 of 106 (9.4%) sera tested from ill, privately owned dogs living in tick-infested areas of Connecticut and New York state had antibodies to E equi, a member of the E phagocytophila genogroup. Titration end points ranged from 1:80 to 1:1,280. Immunoblots revealed antibodies to proteins of E equi having molecular masses of predominantly 29, 40, 44, 105, 120, and 160 kd. There was good agreement between results of serologic testing methods, but use of the human isolate (NCH-1 strain) in western blot analyses detected 2 additional seropositive dogs found to be negative by indirect fluorescent antibody staining methods with the MRK strain.

Clinical Implications

Dogs living in areas where Ixodes scapularis is abundant may be exposed to multiple pathogens, such as E equi or Borrelia burgdorferi. Although mild or subclinical infections with E equi may develop, dogs with marked leukopenia, thrombocytopenia, or anemia should be viewed as possibly having ehrlichiosis. Laboratory diagnosis should include examinations for morulae in granulocytes or monocytes in addition to serologic analyses. (J Am Vet Med Assoc 1997;211:1134–1137)

Free access
in Journal of the American Veterinary Medical Association