Rhipicephalus bursa is a common tick parasite of small-to-medium size ungulates, principally in warm, temperate, and subtropical areas. Although common in livestock and showing a wide geographic distribution, its epidemiological role in tick-borne bacterial disease is barely known. This study addressed the knowledge gap and aimed to screen for the presence of Anaplasmataceae and spotted fever group (SFG) Rickettsia species in R. bursa ticks collected from domestic animals in Romania, Eastern Europe. A total of 64 pools of R. bursa ticks collected from small ungulates were tested by PCR for Anaplasmataceae DNA presence using group-specific primers. Specific testing was performed for Anaplasma marginale/A. centrale/A. ovis, A. platys, A. phagocytophilum, Ehrlichia canis, and SFG Rickettsia. The positive samples were purified and sequenced, and sequences analysis was used to identify the species and to confirm the PCR results. The only pathogen identified in this study was E. canis. The obtained sequences confirmed the PCR results. The presence of E. canis in R. bursa in Romania and in ticks from sheep was shown for the first time in this study. Based on these findings, it may be presumed that the E. canis DNA originated from ticks; however, the vectorial role of R. bursa (and other arthropod species) in the transmission of E. canis should be proved experimentally.

Indirect immunofluorescent antibody (IFA), latex agglutination (IA), and enzyme immunoassay (EIA) methods were compared for evaluation of the serum antibody responses of dogs experimentally and naturally exposed to spotted fever-group rickettsiae. Selected sera (obtained on days 1, 42, 53, 124, 145, 236, 255, 264, and 292) were examined from three 8-month-old female Beagles inoculated with Rickettsia rickettsii on days 34 and 250 of the study. A second group of dogs comprised three 8-month-old female Beagles inoculated with R montana on days 34 and 102. Subsequently, these dogs were inoculated with R rickettsii on day 250. Serum samples were obtained from the second group of dogs on days 1, 96, 103, 132, 180, 215, 292, and 494. A third group consisted of 21 naturally exposed dogs, from which sequentially obtained serum samples were available, and which had clinical signs compatible with Rocky Mountain spotted fever. Clinical signs of disease in dogs of the third group resolved after treatment with tetracycline (22 mg/kg of body weight, Po, q 8 h) was instituted. At least 2 sequentially obtained serum samples from each dog were tested. In general, the first sample was obtained just prior to treatment and the convalescent serum samples were obtained at weekly or greater intervals thereafter. For correlation and reactivity data, an IFA test for IgG/IgM (using heavy and light chains-specific conjugate) was used as the reference standard for comparison of results with those of the other tests,

The vascular permeability of the ocular fundus, alterations in the coagulation system, and plasma concentrations of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) were studied in dogs following intradermal inoculation with 5 x 10(5) TCID50 of Rickettsia rickettsii. Twenty-four to 48 hours after the onset of fever and rickettsemia, multifocal areas of retinal vasculitis were evident, which corresponded to areas of altered vascular permeability demonstrated by fluorescein angiography. The number and intensity of retinal vessels with sodium fluorescein leakage peaked during the second week after inoculation, and retinal vascular permeability remained altered during the third week of infection, well past the phase of clinical and clinicopathologic recovery. Development of retinal vasculitic foci was associated with thrombocytopenia, increased concentrations of circulating fibrinogen, and slight prolongation of activated partial thromboplastin time. Increased concentrations of fibrin/fibrinogen degradation products were detected in 4 of 9 dogs. Despite the degree of vascular endothelial damage evident on fluorescein angiographic and histologic studies in these dogs, plasma TXB2 and 6-keto-PGF1 alpha concentrations were not increased.

Amblyomma hebraeum, 60CO irradiation of nymphs failed to attenuate Cowdria ruminatium

The kinetics of specific IgM and IgG antibody response was characterized in four 9-month-old Beagles after inoculation of 2 x 10(2) plaque-forming units (PFU) of Sheila Smith strain of Rickettsia rickettsii. Immunoglobulin M antibodies were first detected by indirect immunoflorescence on postinoculation (PI) day 9, peaked by PI day 20, and were no longer detectable by PI day 80. Immunoglobulin G antibodies became detectable between PI days 22 and 28, peaked by PI day 42, and decreased gradually through PI day 130. Subsequent challenges with R rickettsii on PI days 216 (2 x 10(2) PFU/dog) and 1,029 (5 x 10(4) tissue culture infective dose [TCID50]/dog) resulted in slightly different serologic responses. The initial challenge exposure failed to increase the concentration of IgG antibodies and induced only low concentrations of IgM antibodies. After the second challenge inoculation, IgM antibodies were not detectable and the concentration IgG antibodies increased slightly. Clinical abnormalities and seroconversion were documented in control dogs following each challenge exposure. Examination of acute and convalescent serum samples from 55 dogs in which Rocky Mountain spotted fever was suspected clinically suggested that sole evaluation of IgM antibodies in acute-phase serum would result in inaccurate diagnoses because of false-positive and -negative results. Use of a composite conjugate that detects IgM and IgG antibodies to R rickettsii appears to be satisfactory for diagnostic purposes; however, concurrent quantitation of IgM antibodies may facilitate serodiagnosis in a select group of dogs in which a four-fold increase in convalescent antibody titer is not detected by use of the composite conjugate. With the exception of a dog with a serum antibody titer of 1:8,192, we were unable to detect IgM or IgG antibodies in CSF samples from 9 dogs with experimentally and 3 dogs with naturally acquired infections.