Antinuclear antibodies can be detected in dog sera reactive to Bartonella vinsonii subsp. berkhoffii, Ehrlichia canis, or Leishmania infantum antigens

The presence of antinuclear antibodies (ANAs) is used to support a clinical diagnosis of systemic lupus erythematosus (SLE) in dogs. However, clinicians must interpret the detection of ANAs with caution, particularly in light of increasing evidence that dogs with known bacterial and protozoal infections can have high ANA titers. Retrospectively, medical records were reviewed for all dogs that were concurrently tested for antinuclear antigens and Bartonella vinsonii (berkhoffii), Ehrlichia canis, or Rickettsia rickettsii antigens between 1990 and 2000. When analyzed on the basis of reactivity to a specific infectious agent, 75% of the B vinsonii (berkhoffii) seroreactors, 16.7% of the E canis seroreactors, and 0% of the R rickettsii seroreactors had concurrent ANAs. Subsequent prospective testing did not detect ANAs in convalescent sera from dogs experimentally infected with B vinsonii (berkhoffii), E canis, or R rickettsii. However, 10-20% B vinsonii (berkhoffii), E canis, or Leishmania infantum reactive sera from naturally infected dogs contained ANAs. In addition, 45% of sera from dogs that are reactive to multiple vectorborne organisms were more likely to contain ANAs when compared to sera from dogs reactive to only 1 test antigen. When interpreting the relevance of seroreactivity to nuclear antigens, clinicians should recognize that dogs with seroreactivity to B vinsonii (berkhoffii), E canis, or L infantum antigens (especially those with seroreactivity to more than one of these pathogens) may produce ANAs.     

A serological survey of Ehrlichia canis, Ehrlichia equi, Rickettsia rickettsii, and Borrelia burgdorferi in dogs in Oklahoma

Serum samples from 259 dogs were tested for antibodies to Ehrlichia canis, Ehrlichia equi, Rickettsia rickettsii, and Borrelia burgdorferi using the indirect fluorescent antibody test. The sera were obtained from submissions to the Oklahoma Animal Disease Diagnostic Laboratory during a 14-month period from June 1986 through July 1987. The rate for positive antibody titers to E. canis was 53%, to E. equi was 33%, to R. rickettsii was 38%, and to B. burgdorferi was 18%. Higher percentages of sera serologically positive to E. canis were found in the spring through the fall months, but there were no seasonal variations for E. equi, R. rickettsii, and B. burgdorferi. There was no consistent pattern of titers to the 4 antigens when age-groups of the dogs were compared. Forty-four different breeds were tested.     

Bartonella henselae IgG antibodies are prevalent in dogs from southeastern USA

In contrast to the large body of literature regarding Bartonella henselae in humans and cats, there is little information about B. henselae as an infectious agent in dogs. Due to the paucity of information regarding the B. henselae serology in dogs, we performed a cross-sectional serosurvey using B. henselae antigen in order to compare the seroprevalence between sick and healthy dogs from the south-eastern USA. Ninety-nine sera were collected from clinically healthy dogs. Three hundred and one sera from sick dogs were submitted to North Carolina State University for serologic screening against a panel of arthropod-transmitted organisms. Serological tests were performed using B. henselae (Bh), Rickettsia rickettsii (Rr), Ehrlichia canis (Ec), Bartonella vinsonii subspecies berkhoffii (Bvb), Babesia canis (Bc) and Borrelia burgdorferi (Bb) antigens. Serum B. henselae IgG antibodies were detected in 10.1% of healthy dogs and in 27.2% of sick dogs. The difference in seroprevalence between the two groups was statistically significant. The majority of seroreactive dogs (80%) had low titers of 1:64 or 1:128. In healthy dogs, seroprevalence for Rr was 14.1% and for Bvb was 1%. In sick dogs, Rr seroprevalence was 29.7%, Ec 6.5%, Bvb 4.7%, Bb 1.7% and Bc was 0.85%. Of the sick dogs that were seroreactive to B. henselae antigens, 40.6% were also seroreactive to Rr, 15.0% reactive to Bvb antigens, 14.8% reactive to Ec antigens, 1.8% reactive to Bc antigens and 1.75% reactive to Bb antigens. Sera from dogs experimentally infected with B. vinsonii subsp. berkhoffii, E. canis or R. rickettsii did not cross react with B. henselae antigens, by IFA testing. This study indicates that B. henselae IgG antibodies are prevalent in healthy and sick dogs living in the south-eastern USA. Nevertheless, further studies are needed to evaluate the epidemiological, clinical and zoonotic relevance of B. henselae infection in dogs.     

A survey for infection with Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi, and Babesia canis in feral and client-owned dogs in the Turks and Caicos Islands, British West Indies

The frequency of infection with Dirofilaria immitis and Babesia canis and seropositivity to Ehrlichia canis and Borrelia burgdorferi in feral and client-owned dogs was determined. Feral dogs were 14.8 and 11.2 times more likely to be seropositive to D. immitis and E. canis, respectively, than were client-owned dogs. None of the dogs tested positive for B. burgdorferi or B. canis.     

Serologic and molecular evidence of coinfection with multiple vector-borne pathogens in dogs from Thailand

Forty-nine dogs from Thailand were evaluated for serologic evidence of exposure or polymerase chain reaction (PCR) evidence of infection with vectorborne pathogens, including Ehrlichia sp. (Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia equi, and Ehrlichia risticii), Bartonella vinsonii subsp. berkhoffi (Bvb), spotted fever group (SFG) rickettsiae (Rickettsia rickettsii), Typhus group (TG) rickettsiae (Rickettsia canada, Rickettsia prowazekii, and Rickettsia typhi), and Babesia sp. (Babesia canis and Babesia gibsonii). All study dogs had at least 1 of 3 entry criteria: fever, anemia, or thrombocytopenia. By immunofluorescence antibody (IFA) testing, seroreactivity was most prevalent to E chaffeensis (74%) and E canis (71%) antigens, followed by E equi (58%), Bvb (38%), E risticii (38%), R prowazekii (24%), B canis (20%), R rickettsii (12%), R canada (4%), and B gibsonii (4%) antigens. There was 100% concordance between E canis IFA and Western blot immunoassay (WI) for 35 of 35 samples; 2 samples were IFA and WI reactive only to E equi antigens. By PCR amplification, 10 dogs were found to be infected with E canis, 5 with Ehrlichia platys, and 3 with B canis. Sequencing of PCR products was undertaken to compare Ehrlichia strains from Thailand to strains originating from the United States. Partial DNA sequence analysis confirmed infection with E canis and E platys, with identical 16S rRNA sequence alignment to E canis (U26740) and to E platys (M83801), as reported in GenBank. Partial E canis P28.1 and P28.2 amino acid sequences from Thai dogs were divergent from analogous sequences derived from North American E canis (AF082744) strains, suggesting that the Thai dogs were infected with a geographically distinct strain of E canis compared to North American strains. The results of this study indicate that dogs in Thailand have substantial exposure to vectorborne diseases and that coinfection with these pathogens may be common.     

Is the detection of anti-Rhipicephalus sanguineus (Rs24p) antibodies a valuable epidemiological tool of tick infestation in dogs?

Antibodies against the 24 kDa Rhipicephalus sanguineus (Rs24p) protein were detected by ELISA to evaluate the relationship between antibodies and tick infestation. The mean titer of 3 dogs that underwent 2 experimental infestations with adult ticks was transiently increased after the second infestation. There was a significant difference in mean titers between positive control dogs naturally infested with ticks and tick-naive dogs. These results suggested that anti-Rs24p antibodies detected by ELISA are a marker of tick exposure. There was no significant difference in mean titers between tick-naive dogs and seropositive dogs to Ehrlichia canis. Some dogs positive for E. canis antibodies showed, however, higher titers than most tick-naive dogs. R. sanguineus may be related to the E. canis infection in Japan.     

Susceptibility and reservoir potential of the dog to spotted fever-group rickettsiae

This study was initiated to determine the degree of susceptibility of dogs to virulent and nonvirulent spotted fever-group rickettsiae and to evaluate dogs as sources of infection for ticks. Dogs were exposed either by inoculation (syringe) or by infective tick bite to the following rickettsial serotypes: (1) Rickettsia rickettsii (Wachsmuth and Sawtooth female 2 strains), (2) R montana (M/5-6 B strain), and (3) R rhipicephali (3-7-female 6 strain). Results indicated that dogs inoculated with 1,000 or 10,000 egg infective doses of virulent R rickettsii developed a rickettsemia that was detectable as early as 4 days after inoculation to as late as 10 days. Conversely, none of the dogs inoculated with R montana (M/5-6 B) or R rhipicephali (3-7-female 6) or exposed to ticks infected with these strains developed detectable rickettsemia, fever, or other observable clinical signs. None of the 394 ticks that fed on rickettsemic dogs (R rickettsii) infected by inoculation became infected, and only 3 of 348 ticks (0.9% infection rate) were infected after feeding on dogs which had been infected by tick bite. All ticks fed on dogs exposed to R rhipicephali and R montana were shown to be free of rickettsiae. The largest concentrations of plaque-forming units (PFU) in Vero cell culture from undiluted whole blood were found on day 6 and on day 7 in dogs that were inoculated with 10,000 and 1,000 R rickettsii, respectively, of the Sawtooth female 2 strain. The highest rickettsial concentration observed for the dog infected by tick feeding was on day 9.(ABSTRACT TRUNCATED AT 250 WORDS)     

The immunologic response of dogs to Bartonella vinsonii subspecies berkhoffii antigens: as assessed by Western immunoblot analysis.

Bartonella vinsonii subspecies berkhoffii is a recently recognized zoonotic pathogen that causes endocarditis, granulomatous rhinitis, and granulomatous lymphadenitis in dogs. Isolation of B. vinsonii (berkhoffii) from blood or tissue samples is frequently unsuccessful; therefore, diagnosis is primarily dependent on serologic or molecular testing modalities. Because previous canine serologic studies have used an indirect immunofluorescence assay (IFA), without Western immunoblot (WI) confirmation, the overall objective of this study was to examine the diagnostic use of WI for confirmation of B. vinsonii (berkhoffii) infection in dogs. To confirm that agar-grown and cell culture-grown organisms yielded similar patterns of WI antigenic protein recognition, the 2 preparations were compared using IFA-reactive sera obtained from dogs experimentally infected with B. vinsonii (berkhoffii). Temporal changes in the pattern of antigenic protein recognition were characterized using sera obtained from dogs at various time points after experimental B. vinsonii (berkhoffii) infection. The specificity of B. vinsonii (berkhoffii) WI was examined by testing canine sera that were reactive to B. henselae, B. clarridgeiae, Ehrlichia canis, Rickettsia rickettsii, Babesia canis, Anaplasma phagocytophilum (previously E. equi), or Brucella canis antigens. Clinical accessions including serum samples obtained from B. vinsonii (berkhoffii) culture-positive dogs and B. vinsonii (berkhoffii) culture-negative dogs that were IFA seroreactive to B. vinsonii (berkhoffii) antigens were examined by WI. The results of this study indicate that WI using agar-grown or cell culture-grown B. vinsonii (berkhoffii) antigens produce identical patterns of antigenic protein recognition. After experimental infection, there is a progressive increase in the number of antigenic proteins that are recognized by WI, with the 33-kD antigen representing the first and the most persistent antigen recognized by B. vinsonii (berkhoffii)-infected dogs. Regarding specificity, sera from dogs that were reactive to various heterologous antigens did not recognize B. vinsonii (berkhoffii) antigens by IFA or WI, and sera from dogs experimentally infected with B. henselae did not recognize B. vinsonii (berkhoffii) antigens by WI. Regarding clinical accessions, there was good agreement between B. vinsonii (berkhoffii) IFA test results and WI analysis. Western immunoblot analysis can be used to detect or confirm exposure to B. vinsonii (berkhoffii) in dogs.     

Seroprevalence of spotted fever group rickettsiae in canines along the United States–Mexico border

Portions of northern Mexico are experiencing a re‐emergence of Rocky Mountain spotted fever (RMSF), a tickborne disease caused by Rickettsia rickettsii, a member of the spotted fever group of rickettsiae (SFGR). Infection with R. rickettsii can result in serious and life‐threatening illness in people and dogs. Canine seroprevalence has been used as a sentinel for human RMSF in previous studies. This study aims to quantify SFGR seroprevalence in canines in three northern Mexican states and identify risk factors associated with seropositivity. A total of 1,136 serum samples and 942 ticks were obtained from dogs participating in government sterilization campaigns and from animal control facilities in 14 Mexican cities in three states. SFGR antibodies were detected using indirect immunofluorescence antibody assays at titre values ≥1/64. Six per cent (69 dogs) showed antibodies to SFGR, with the highest seroprevalence reported in Baja California (12%), Coahuila (4%) and Sonora (4%). Dogs from Baja California had three times higher odds of having SFGR antibodies compared to dogs from Sonora (OR = 3.38, 95% CI, 1.81–6.37). Roughly one quarter (25%) of surveyed dogs were parasitized by ticks (Rhipicephalus sanguineus sensu lato) at the time of sample collection. A portion of collected ticks were tested for rickettsial DNA using polymerase chain reaction. Positive samples were then sequenced, showing evidence of SFGR including R. massiliae, R. parkeri and R. rickettsii. Dogs that spent the majority of time on the street, such as free‐roaming or community‐owned dogs, showed a greater risk of tick infestation, seropositivity, bearing seropositive ticks, and may play a pivotal role in the spread of SFGR among communities. Estimating the seroprevalence of SFGR in the canine population can help public health campaigns target high‐risk communities for interventions to reduce human RMSF cases.     

Seroprevalence of antibodies against Bartonella species and evaluation of risk factors and clinical signs associated with seropositivity in dogs

OBJECTIVE: To determine the seroprevalence of antibodies against Bartonella spp in a population of sick dogs from northern California and identify potential risk factors and clinical signs associated with seropositivity. SAMPLE POPULATION: Sera from 3,417 dogs. PROCEDURE: Via an ELISA, sera were analyzed for antibodies against Bartonella vinsonii subsp berkhoffii, Bartonella clarridgeiae, and Bartonella henselae; test results were used to classify dogs as seropositive (mean optical density value > or = 0.350 for B henselae or > or = 0.300 for B clarridgeiae or B vinsonii subsp berkhoffi) or seronegative. Overall, 305 dogs (102 seropositive and 203 seronegative dogs) were included in a matched case-control study. RESULTS: 102 of 3,417 (2.99%) dogs were seropositive for > or = 1 species of Bartonella. Of these, 36 (35.3%) had antibodies against B henselae only, 34 (33.3%) had antibodies against B clarridgeiae only, 2 (2.0%) had antibodies against B vinsonii subsp berkhoffii only, and 30 (29.4%) had antibodies against a combination of those antigens. Compared with seronegative dogs, seropositive dogs were more likely to be herding dogs and to be female, whereas toy dogs were less likely to be seropositive. Seropositive dogs were also more likely to be lame or have arthritis-related lameness, nasal discharge or epistaxis, or splenomegaly. CONCLUSIONS AND CLINICAL RELEVANCE: Only a small percentage of dogs from which serum samples were obtained had antibodies against Bartonella spp. Breed appeared to be an important risk factor for seropositivity. Bartonella infection should be considered in dogs with clinical signs of lameness, arthritis-related lameness, nasal discharge or epistaxis, or splenomegaly.     

Seroepidemiology of Bartonella vinsonii subsp berkhoffii exposure among healthy dogs

OBJECTIVE: To determine seroprevalence of antibodies to Bartonella vinsonii subsp berkhoffii and risk factors for seropositivity among working dogs owned by the US government. DESIGN: Cross-sectional study. ANIMALS: 1,872 dogs. PROCEDURE: An ELISA was used to detect antibodies to B vinsonii subsp berkhoffii. RESULTS: Antibodies to B vinsonii subsp berkhoffii were detected in 162 dogs (8.7%; 95% confidence interval, 7.4 to 10.0%). Dogs living in the southeast, plains states, southwest, and south-central were significantly more likely to be seropositive than were dogs living in other regions of the United States. German Shepherd-type dogs were significantly less likely to be seropositive than were dogs of other breeds, and dogs entering training programs or that had been rejected from a training program were significantly more likely to be seropositive than were dogs used for narcotics detection and dogs trained to patrol or detect explosives. Dogs used by the border patrol or Federal Aviation Administration were more likely to be seropositive than were dogs used by the Department of Defense or customs service. Odds that dogs would be seropositive were significantly higher for dogs stationed in the southern United States, the northeastern United States, or a foreign country, compared with dogs stationed in all other regions of the United States. CONCLUSIONS AND CLINICAL RELEVANCE: Overall, 8.7% of this diverse group of healthy dogs was found to be seropositive for antibodies to B vinsonii subsp berkhoffii, and seropositivity rates were associated with location, suggesting either that there are multiple vectors for the organism or that the major vector for the organism depends on geographic and environmental factors.     

Infection by spotted fever rickettsiae in people, dogs, horses and ticks in Londrina, Parana State, Brazil

Spotted fever is a disease caused by bacteria from the genus Rickettsia of the spotted fever group (SFG). Rickettsia rickettsii is likely the main agent of Brazilian spotted fever (BSF). With the objective of gathering information on the circulation of SFG rickettsiae in Londrina, Parana state, ticks from dogs and horses and also blood from dogs, horses and humans were collected in a neighbourhood of the city which presented potential for circulation of rickettsiae between hosts and vectors. Amblyomma cajennense, Dermacentor nitens, and Rhipicephalus sanguineus ticks were subjected to Polymerase Chain Reaction targeting a fragment of the Rickettsia gltA gene. This specific gene encodes the enzyme citrate synthase of Rickettsia spp., and results on all ticks were negative. Human and animal sera were tested by Indirect Immunofluorescence Assay in which R. rickettsii and R. parkeri were used as antigens. Sera from 4.7% human, 2.7% canine and 38.5% equine were positive for R. rickettsii. For R. parkeri, 0.9% human, 2.7% canine and 11.5% equine samples were positive. All samples reactive to R. parkeri also reacted to R. rickettsii. An epidemiological questionnaire was applied, but there were no statistically significant results. Comparison of our serological results with previous studies in Brazil, among BSF endemic and non-endemic areas, indicates that there is no established rickettsial infection in the study area, a statement corroborated with our molecular analysis. Nonetheless, as humans of the present study are highly exposed to tick infestations, health education within the population is needed to obtain efficient tick control.     

Transstadial and intrastadial experimental transmission of Ehrlichia canis by male Rhipicephalus sanguineus

The acquisition and transmission of rickettsial pathogens by different tick developmental stages has important epidemiological implications. The purpose of this study was to determine if male Rhipicephalus sanguineus can experimentally acquire and transmit Ehrlichia canis in the absence of female ticks. Two trials were performed where nymphal and male R. sanguineus were simultaneously acquisition fed on the same infected donor hosts, and transstadially or intrastadially exposed male ticks were fed on separate pathogen-free dogs as a test for transmission. A single-step p30-based PCR assay was used to test canine and tick hosts for E. canis infections before and after tick feeding. E. canis was detected after either intrastadial or transstadial passage in male ticks, the organism remained detectable in both tick groups after transmission feeding, and both tick groups transmitted the rickettsia to susceptible dogs. Infection of dogs via tick feeding resulted in milder clinical signs and lower antibody titers than intravenous inoculation of carrier blood, but further investigation is needed to understand the mechanisms responsible for this observation. These results demonstrate that male R. sanguineus can take multiple feedings, and that they can both acquire and transmit E. canis in the absence of female ticks. This tick development stage could be important in transmission of E. canis, and perhaps related pathogens, between vertebrate hosts under natural and experimental conditions.     

The low seroprevalence of tick-transmitted agents of disease in dogs from southern Ontario and Quebec

Infectious diseases caused by pathogens transmitted by ticks and other insect vectors are an important cause of morbidity and mortality in both dogs and humans throughout North America. The purpose of this study was to determine the seroprevalence of selected vector-transmitted pathogens in southern Ontario and Quebec. Samples submitted to the Vector Borne Disease Diagnostic Laboratory (VBDDL) at the North Carolina State University College of Veterinary Medicine were evaluated for antibodies to Ehrlichia canis, Anaplasma phagocytophilum, Babesia canis, Bartonella henselae, Borrelia burgdorferi, Bartonella vinsonii subspecies berkhoffii, and Rickettsia rickettsii. Information regarding breed and the city or province from which the sample originated was recorded; however, travel history was unknown for the majority of dogs. Overall seroprevalence to these tick-borne pathogens in southern Ontario and Quebec is low compared with most regions of the United States, suggesting that veterinarians in this region of Canada should pursue diagnostic evidence of infection in dogs with a travel history or prior residence in areas endemic for exposure to tick-borne infections.     

Clinical and serologic studies of canine borreliosis

During 1984 and 1985, blood samples were obtained from 271 dogs that were suspected of having borreliosis. The dogs lived in areas known to be infested with ticks and had been examined because of limb/joint disorders or for unknown illnesses marked by fever, anorexia, or fatigue. Lameness had been the most frequently reported clinical manifestation. Analyses of serum specimens, by an indirect fluorescent antibody (IFA) method or by an ELISA, detected antibodies to Borrelia burgdorferi, the etiologic agent of borreliosis in dogs and of Lyme disease in human beings. Antibody to B burgdorferi was detected in 76.3% of 114 specimens from dogs living in the lower Hudson Valley region of New York State (predominantly Westchester County), in 66.5% of 155 specimens from dogs from southern Connecticut, and in single specimens from dogs from Rhode Island and California. Geometric mean antibody titers peaked during the winter. Results of IFA tests and ELISA were in agreement, but the latter method yielded less variable results, had greater sensitivity, and was more easily standardized. Five dogs from New York State and Connecticut seropositive to B burgdorferi had developed kidney disorders during or after episodes of intermittent lameness. Application of murine monoclonal antibody in an IFA procedure verified the presence of B burgdorferi in renal cortical tissues from one dog.     

Evaluation of Rhipicephalus sanguineus as a potential biologic vector of Ehrlichia platys

The brown dog tick, Rhipicephalus sanguineus (Acari: Ixodidae), transmits several diseases among dogs including Ehrlichia canis infection. The role of Rhipicephalus sanguineus as a biologic vector for E platys, the rickettsial agent of infectious canine cyclic thrombocytopenia, was studied in dogs. Laboratory-cultured, pathogen-free nymph ticks were fed to repletion on dogs acutely infected with E platys. Tick engorgement coincided with the development of initial parasitemia and thrombocytopenia in the infected dogs. Following repletion, nymph ticks were allowed to molt under controlled conditions. One-month-old E platys-exposed adult ticks failed to infect naive dogs in animal transmission studies. The presence of E platys was not detected in midguts or salivary glands of similarly exposed adult ticks by use of light and transmission electron microscopy. These studies indicate that R sanguineus may not transmit E platys infection.     

Serologic evidence of Rickettsia akari infection among dogs in a metropolitan city

OBJECTIVE: To determine whether dogs in New York, NY are naturally infected with Rickettsia akari, the causative agent of rickettsialpox in humans. DESIGN: Serologic survey. ANIMALS: 311 dogs. PROCEDURE: Serum samples were obtained from dogs as a part of a study on Rocky Mountain spotted fever and borreliosis or when dogs were examined at area veterinary clinics for routine care. Dog owners were asked to complete a questionnaire inquiring about possible risk factors at the time serum samples were obtained. Samples were tested for reactivity to spotted fever group rickettsiae by use of an enzyme immunoassay (EIA). Twenty-two samples for which results were positive were tested by use of an indirect immunofluorescence antibody (IFA) assay followed by confirmatory cross-absorption testing. RESULTS: Results of the EIA were positive for 24 (7.7%) dogs. A history of tick infestation and increasing age were significantly associated with whether dogs were seropositive. Distribution of seropositive dogs was focal. Seventeen of the 22 samples submitted for IFA testing had titers to R rickettsii and R akari; for 11 of these, titers to R akari were higher than titers to R rickettsii. Cross-absorption testing indicated that in 6 of 7 samples, infection was caused by R akari. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dogs can be naturally infected with R akari. Further studies are needed to determine the incidence of R akari infection in dogs, whether infection is associated with clinical illness, and whether dogs can serve as sentinels for human disease.     

Cyclic CD8+ lymphopenia in dogs experimentally infected with Bartonella vinsonii subsp. berkhoffii

Until recently, it was presumed that Bartonella vinsonii only infected voles, a species of North American rodents. In April of 1993, however, our laboratory isolated a novel subspecies of B. vinsonii (B. vinsonii subsp. berkhoffii) from the blood of a dog diagnosed with vegetative valvular endocarditis. Subsequently, based on a seroepidemiologic survey of dogs from North Carolina and Virginia presenting for a variety of medical problems, we found evidence supporting a potentially important association between B. vinsonii and Ehrlichia canis co-infection in dogs. In the following study, eight dogs were infected with B. vinsonii: four specific pathogen free dogs and four dogs that had previously been infected with E. canis. Flow cytometric analysis of peripheral blood lymphocytes revealed a cyclic elevation of the CD4/CD8 T-cell ratio that correlated with cyclic CD8+ lymphopenia in all dogs infected with B. vinsonii, regardless of prior exposure to E. canis.     

Prevalence of Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, Bartonella vinsonii berkhoffii, and Rickettsia spp. in dogs from Grenada

To identify the tick-borne pathogens in dogs from Grenada, we conducted a serologic survey for Ehrlichia canis in 2004 (104 dogs) and a comprehensive serologic and molecular survey for a variety of tick-borne pathogens in 2006 (73 dogs). In 2004 and 2006, 44 and 32 dogs (42.3% and 43.8%) were seropositive for E. canis, respectively. In 2006, several tick-borne pathogens were identified by serology and PCR. DNA of E. canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, and Bartonella sp. were identified in 18 (24.7%), 14 (19.2%), 5 (7%), 5 (7%), and 1 (1.4%) dogs, respectively. Six (8.2%) dogs were seropositive for Bartonella vinsonii subsp. berkhoffii. All dogs were seronegative and PCR-negative for Rickettsia spp. Coinfection with two or three pathogens was observed in eight dogs. Partial 16S rRNA E. canis and A. platys sequences were identical to sequences in GenBank. Partial 18S rRNA gene sequences from the Grenadian H. canis were identical to each other and had one possible mismatch (ambiguous base) from H. canis detected from Spain and Brazil. Grenadian B. c. vogeli sequences were identical to B. c. vogeli from Brazil and Japan. All of the detected pathogens are transmitted, or suspected to be transmitted, by Rhipicephalus sanguineus. Results of this study indicate that dogs from Grenada are infected with multiple tick-borne pathogens; therefore, tick-borne diseases should be included as differentials for dogs exhibiting thrombocytopenia, leukopenia, fever, or lethargy. One pathogen, E. canis, is also of potential public health significance.     

Spirochetemia caused by Borrelia turicatae infection in 3 dogs in Texas

Spirochetemia was diagnosed in 2 Siberian Huskies and a Rottweiler from the northwestern region of Texas between June 1999 and October 2001. Clinical findings were nonspecific; tick exposure was documented in 2 of the dogs. Hematologic abnormalities included anemia (n=2), neutrophilia (n=2, including 1 with a left shift), lymphopenia (n=3), eosinopenia (n=3), and thrombocytopenia (n=2). One anemic dog had a positive Coombs' test. In 1 dog, Western blot analysis of serum yielded multiple positive bands with B turicatae lysate, indicating the spirochetemia most likely was due to B turicatae infection. In 2 dogs, spirochetes were cultured from the blood and identified using DNA analysis as Borrelia turicatae; 1 of these dogs also was seropositive for Ehrlichia canis and B burgdorferi. In 2 cases, spirochetemia was more prominent in blood smears prepared immediately after sample collection than in smears prepared from EDTA blood. Two dogs recovered with doxycycline treatment; 1 dog declined clinically despite treatment and was euthanized. B turicatae is the agent of tick-borne (endemic) relapsing fever in humans and is distinct from B burgdorferi, the agent of Lyme disease; however, serologic cross-reactivity may occur. B turicatae is transmitted by the soft tick, Ornithodoros turicata, and infection should be considered in dogs with spirochetemia and possible exposure to the tick vector.