Antibodies to spotted fever-group rickettsiae in dogs and prevalence of infected ticks in southern Connecticut

Blood samples and ticks were obtained from dogs to assess canine exposure to spotted fever-group (SFG) rickettsiae during 1978-1980 in southern Connecticut. Of the 1,576 dog sera screened by microimmunofluorescence. 174 (11.0%) contained specific antibodies at titers greater than or equal to 1:64 against Rickettsia montana (n = 34), R rickettsii (n = 31), R rhipicephali (n = 19), or the unclassified 369-C rickettsia (n = 90). End points greater than or equal to 1:8,192 to R rickettsii and to R rhipicephali were recorded for 6 and 3 sera, respectively. Seropositivity rates from southwestern and southeastern Connecticut were similar (about 11%), with positive sera obtained from each region in nearly all months of the investigation. Rates were between 10% for dogs 2 to 7 years old and 14% for those greater than or equal to 8 years. Eight of 629 Dermacentor variabilis, 1 of 18 Ixodes dammini, and 2 of 3 Amblyomma americanum were positive by direct immunofluorescence for SFG rickettsiae. Thirteen D variabilis contained unidentified, long, bacillus-like organisms that differed from the short, ovoid (coccal) forms typical fo the spotted-fever agent, R rickettsii. With the exposure to infected ticks and production of type-specific antibodies against at least 4 SFG antigens, dogs may serve as suitable enzootic or epizootic indicators of rickettsial activity.     

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)     

Antibodies to Borrelia burgdorferi in dogs in North Carolina

An indirect immunofluorescence assay was used to detect antibodies against Borrelia burgdorferi in sera from 600 dogs in 1983 and 402 dogs in 1985. In 1983, the overall prevalence rate of dogs with B burgdorferi titers greater than or equal to 1:64 was 3.6%, whereas in 1985, the prevalence rate was 2.7%. An unexplainable higher seroprevalence was detected in 1 group of dogs tested in 1983. These dogs were from the southern coastal plains of North Carolina. In the dogs tested in 1985, this regional difference in sero-prevalence was not noticed. Statistical differences were not noticed (P greater than 0.05) between dogs from 2 sources or when gender was considered. Seemingly, the prevalence of anti-B burgdorferi antibodies in dogs in North Carolina was low.     

Antibody activity to Babesia canis in dogs in North Carolina

Sera from 600 dogs from 6 geographic regions of North Carolina were analyzed for antibody activity to Babesia canis, using an indirect fluorescent antibody test. Overall, 3.8% of the dogs had an antibody titer greater than or equal to 1:80, which was considered to be indicative of infection. Dogs were more likely to be seropositive if they were housed at humane facilities and were located within areas of the state with a milder climate than were pet animals living within colder regions of North Carolina.     

Babesia gibsoni infections in dogs from North Carolina

The recognition of canine babesiosis in North Carolina caused by Babesia gibsoni documents the expansion of the previously reported endemic area of this disease. Clinical signs ranged from severe hemolytic anemia and thrombocytopenia to subclinical infections. No infected dogs had traveled to endemic areas. Antibabesial treatment failed to eradicate the organism from infected dogs.     

Identification of rickettsiae in cutaneous biopsy specimens from dogs with experimental Rocky Mountain spotted fever

Direct immunofluorescence reaction for Rickettsia rickettsii was performed on formalin-fixed, paraffin-embedded cutaneous biopsy specimens collected from dogs with experimental Rocky Mountain spotted fever (RMSF). A technique of trypsin digestion of deparaffinized, rehydrated sections was successful in demonstrating discrete, immunofluorescent organisms in endothelia and adjacent vessel walls in the dermis. R rickettsii was identified only in grossly evident dermal lesions (macular rash or oral vesicles) and was not apparent in randomly collected biopsy specimens from clinically normal inguinal skin. These results suggest that clinical application of this technique for diagnosis of RMSF may be limited in dogs without cutaneous lesions.     

Serological prevalence study of exposure of cats and dogs in Launceston, Tasmania, Australia to spotted fever group rickettsiae

A sero-epidemiological study of cats and dogs in the Launceston area of Tasmania, Australia was undertaken to determine the prevalence of antibodies to spotted fever group (SFG) rickettsiae. Results showed that 59% of cats and 57% of dogs were positive for antibodies, but there was no correlation between the animal's health and seropositivity at the time of testing, suggesting that rickettsial exposure is unrelated to ill-health in these two species of domestic animals.     

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.     

In vitro cultivation of a newly recognized Babesia sp. in dogs in North Carolina

A novel large Babesia sp. from an infected dog was cultivated in vitro by microaerophilous stationary phase culture methodology. A primary culture initiated in enriched RPMI-1640 medium supplemented with 40% canine serum and incubated in a 2% oxygen environment supported parasite growth in vitro. Subsequent subcultures into enriched HL-1 medium with 20% fetal bovine serum also supported parasite propagation. Cultures were successfully introduced to 5% carbon dioxide in air atmosphere at passage 4. To date, the parasites have been continuously cultured through 35 passages, although the parasitemias are low, ranging from 0.2 to 0.3%. Parasites cultured in RPMI with canine serum were cryopreserved and successfully recovered from liquid nitrogen storage. The small subunit ribosomal rRNA gene sequence was identical in blood-derived and culture-derived parasites, differing in a single base position from the previously reported sequence for this Babesia sp. The ultrastructure of the parasite was consistent with that of other large Babesia spp., except that the spherical body contained numerous round particles unlike the inclusions previously described in Babesia spp.     

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.     

Using policy goals to evaluate red wolf reintroduction in eastern North Carolina

Despite being politicized in its early stages, the Red Wolf (Canis rufus) Recovery Program (RWRP) existed for 30 years in eastern North Carolina with little attention paid to finding common ground between policy contestants. We situated our data from 62 key informant interviews within Stone’s (2002) policy goals framework to evaluate red wolf recovery in eastern North Carolina, United States. Four goals underpin U.S. political culture and offer common ground with which to begin negotiation among policy contestants. Our results highlight challenges to achieving one definition prescribing how to treat each policy goal. The future of recovery within the red wolf’s historic range could hinge on decision-makers’ willingness to prioritize policy design over policy tool evaluation, developing a process by which to achieve an agreed-upon set of values underscoring red wolf recovery. This process must reconcile disagreements about what is fair or equitable treatment of landowners and include local-level experiences.     

Ocular manifestations of Rocky Mountain spotted fever in dogs

Dogs were examined ophthalmoscopically to determine the prevalence and type of ocular lesions associated with naturally acquired Rocky Mountain spotted fever (RMSF). In a consecutive series of 11 dogs with serologically confirmed RMSF, 9 had ocular involvement reflecting various degrees of vascular damage, including subconjunctival hemorrhage, hyphema, anterior uveitis, iris stromal hemorrhage, retinal petechiae, and focal areas of retinal edema and perivascular inflammatory cell infiltrate. The ophthalmic lesions generally were mild, and most resolved without complication after systemic administration of appropriate antibiotics. The high prevalence of ocular lesions in these dogs suggested that critical ophthalmologic evaluation is a useful diagnostic tool for the early detection and management of RMSF in dogs.     

Serosurvey of antiBabesia antibodies in stray dogs and American pit bull terriers and American staffordshire terriers from North Carolina

Stray dogs (n=359) and kennel dogs (n=149) from North Carolina were tested for evidence of antiBabesia antibodies. AntiBabesia antibodies were detected in 21/359 and 22/149 of the stray and kennel dogs, respectively. A total of 57 dogs from both groups were tested for babesiasis by light microscopy and polymerase chain reaction (PCR). Babesia deoxyribonucleic acid (DNA) was detected in 3/28 of the stray dogs and 14/29 of the kennel dogs. When Babesia DNA was detected by PCR, the species-specific PCR results differed from the Babesia species antibody titer results in 6/17 of the PCR-positive dogs. There was no association between antiBabesia antibodies and the presence of ticks. There are currently Babesia gibsoni epizootics affecting American pit bull terrier kennels.     

Seroprevalence of Ehrlichia canis, Ehrlichia equi, and Ehrlichia risticii in sick dogs from North Carolina and Virginia

Ehrlichia canis, E. equi, and E. risticii seroprevalence was determined by microimmunofluorescent antibody testing (IFA) in a sequential population of 1,845 sick dogs admitted during a 1-year period to the North Carolina State University Veterinary Teaching Hospital. A seroreactor was defined by a reciprocal IFA titer of > or =80 to E. canis, E. equi, or E. risticii antigens. Of the 48 IFA seroreactors, 44 dogs were seroreactive to E. canis, 21 to E. equi, and 0 to E. risticii. Seventeen dogs reacted to both E. canis and E. equi antigens. There was concordance of E. canis IFA and western immunoblot (WI) test results for 36/44 dogs. Because of cross-reactivity of E. canis sera with E. equi antigens, WI was of less utility to confirm E. equi exposure. After elimination of E. canis seroreactors, there was concordance of 2/4 E. equi IFA and WI test results. Based upon a retrospective review of medical records, ehrlichiosis was diagnosed in 10/48 (21%) IFA seroreactive dogs, 9 of which were confirmed positive by WI. Of the remaining 38 IFA seroreactors, 29 also were confirmed by E. canis or E. equi WI. These results indicate that (1) ehrlichiosis was not diagnosed in the majority of serologically confirmed cases, (2) based upon E. canis and E. equi WI analysis, IFA testing was not specific (21% false positive), (3) E. canis sera cross-react with E. equi antigens, and (4) serologic evidence of E. risticii infection was lacking in the dog population studied.     

Granulocytic Ehrlichiosis in Dogs from North Carolina and Virginia

Medical records of 3 dogs from North Carolina and 3 dogs from Virginia with ehrlichial morulae in circulating neutrophils were studied retrospectively. Two clinically distinct disease syndromes, including chronic, moderate to severe anemia (n = 3) and polyarthritis (n = 2) were associated with canine granulocytic ehrlichiosis (CGE) in these dogs. One dog was clinically healthy, and abnormalities were not detected during physical examination. Clinical signs were nonspecific and included fever, lethargy, anorexia, vomiting, and diarrhea. The most frequent laboratory abnormalities were normocytic normochromic nonregenerative anemia, moderate thrombocytopenia with large platelets, lymphopenia, and eosinopenia. Considerable variability was found in the serologic responses to Ehrlichia equi, Ehrlichia canis , and Ehrlichia chaffeensis antigens among the 5 dogs for which stored sera were available for indirect fluorescent antibody testing. Polymerase chain reaction amplification and sequencing of portions of the 16S rRNA gene from blood (collected in ethylenediaminetetraacetic acid) of 1 severely anemic dog (dog 3) and 1 polyarthritic dog (dog 4) resulted in DNA sequences nearly identical to the GenBank accessions for Ehrlichia ewingii. The DNA sequence from a 3rd dog (dog 5) was most similar to that of E. canis. Serologic or molecular results support the possibility of E. ewingii, E. equi , and E. canis coinfection or serologic cross-reactivity among canine granulocytic and monocytic Ehrlichia species in dogs from North Carolina and Virginia. Variability in response to tetracycline or doxycycline treatment was noted in these dogs, with more rapid resolution of signs in dogs with polyarthritis. We report the 1st cases of CGE in dogs from North Carolina and Virginia, including recognition of CGE in a healthy dog.     

Dermal necrosis associated with Rocky Mountain spotted fever in four dogs

Four dogs with Rocky Mountain spotted fever developed extensive dermal necrosis. Factors contributing to this complication included delay in initiation of appropriate therapy.