Assessment of the effect of long-term serum storage for retrospective serologic diagnosis of canine monocytic ehrlichiosis (Ehrlichia canis).

There is currently sparse information on the possible effect of long-term storage of serum specimens for the retrospective serodiagnosis of canine monocytic ehrlichiosis (CME). The aim of this study was to assess the agreement between the original serologic outcome and the results of a repeat indirect fluorescent antibody (IFA) assay for the detection of IgG antibodies against E. canis. A secondary aim was to compare the diagnostic performance of two commercially available point-of-care (POC) immunochromatographic (IC) assays. Archived serum samples originally tested as positive (n=66) or negative (n=19) for E. canis IgG antibodies and kept frozen at -20°C for a median of 22 years, were retrospectively examined by IFA and by two POC IC assays. Cohen's Kappa coefficient (0.748, p < 0.0001), indicated a substantial agreement between the original and repeat serologic testing results. An almost identical high sensitivity and moderate specificity were established for the two POC IC assays. Canine serum specimens on long-term storage may still be of value for seroepidemiologic surveys investigating the exposure to E. canis.

Assessment of a quantitative 5' nuclease real-time polymerase chain reaction using groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil.

New genotypes of Anaplasmataceae agents have been detected in wild carnivores, birds and deer in Brazil. The present work aimed to investigate the presence of Ehrlichia and Anaplasma species in rodents sampled in Brazil. Additionally, a newly designed quantitative 5' nuclease real-time multiplex PCR for Ehrlichia and Anaplasma spp. detection based on groEL gene amplification was designed, showing high specificity and sensitivity (10 groEL fragment copy/µL). Between 2000 and 2011, different rodent species [n=60] were trapped in 5 Brazilian biomes. Among 458 rodent spleen samples, 0.4% (2/458) and 2.4% (11/458) were positive for Ehrlichia and Anaplasma spp., respectively. Of 458 samples, 2.0% (9/458) and 1.1% (5/458) were positive for Anaplasma sp. and Ehrlichia sp., respectively, using conventional 16S rRNA PCR assays. Maximum Likelihood phylogenetic analyse based on a small region of 16S rRNA genes positioned the Anaplasma genotypes in rodents near Anaplasma phagocytophilum or Anaplasma marginale and Anaplasma odocoilei isolates. Ehrlichia genotypes were closely related to E. canis. There was a low occurrence of Anaplasma and Ehrlichia in wild and synanthropic rodents in Brazil, suggesting the circulation of new genotypes of these agents in rodents in the studied areas.

Detection of Anaplasma platys in dogs using real-time loop-mediated isothermal amplification.

Anaplasma platys is a parasite of canine platelets that causes infectious cyclic thrombocytopenia. In this study, a novel real-time loop-mediated isothermal amplification (RT-LAMP) method was developed to detect A. platys. RT-LAMP primer sets were designed using a citrate synthase gene sequence and the assay was performed at 63 °C for 30 min. No cross-reactivity was observed with other Anaplasma or Ehrlichia spp. and the method exhibited a similar level of sensitivity in detecting the organism in 58 canine blood samples to that of a nested PCR. This RT-LAMP is a rapid and potentially cost-effective method of diagnosing A. platys infection in dogs.

Prospective assessment of the etiology of acute febrile illness after a tick bite in Slovenia.

A prospective study established the etiology of febrile illnesses in residents of Slovenia that occurred within 6 weeks after a tick bite. A combination of laboratory and clinical criteria identified 64 (49.2%) of 130 patients as having confirmed, probable, or possible cases of tickborne disease during 1995 and 1996. Of the 130 patients, 36 (27.7%) had laboratory evidence of tickborne encephalitis, all of whom had clinically confirmed disease. Evidence of infection with Borrelia burgdorferi sensu lato was identified in 26 patients; 10 (7.7%) had confirmed Lyme borreliosis. Of 22 patients with evidence of Ehrlichia phagocytophila infection, 4 (3.1%) had confirmed ehrlichiosis. Infection by multiple organisms was found in 19 (14.6%) of 130 patients. Patients with meningeal involvement (43 [72.3%] of 59) were more likely to have confirmed tickborne disease than were patients with illness of undefined localization (18 [26.5%] of 68; P<.0001). Tickborne viral and bacterial infections are an important cause of febrile illness in Slovenia.

Evaluation of vaccination of horses as a strategy to control equine monocytic ehrlichiosis.

OBJECTIVE: To determine whether preferentially vaccinated horses were at risk for exposure to Ehrlichia risticii, whether horses with equine monocytic ehrlichiosis (EME) were likely to have been nonvaccinated, and whether clinical severity and financial costs associated with care and treatment of EME were less for vaccinated horses with EME than for nonvaccinated horses with EME. DESIGN: Cross-sectional and case-control studies. PROCEDURE: Information on usage of E risticii bacterins to control EME was collected for 2,587 horses located on 511 farms throughout New York. Each horse was tested for serum antibodies directed against E risticii. Data on efficacy of vaccination to reduce the prevalence and clinical severity of EME and monetary losses associated with EME were collected from 68 horses with EME and 132 clinically normal horses. RESULTS: A correlation was not detected between the county seropositive proportion and the proportion of horses vaccinated against EME. Among horses diagnosed for EME, median date of diagnosis was not delayed for vaccinated horses, compared with that for nonvaccinated horses. Mean cost per case was not significantly different for nonvaccinated horses, compared with that for vaccinated horses ($ 1,082 and $ 1,001, respectively). Vaccination was not associated with a reduction in prevalence or in severity of EME-related clinical signs. CLINICAL IMPLICATIONS: Administering killed E risticii bacterin once a year to control EME in New York appears to have limited success. Among horses in which EME was diagnosed, severity of illness and financial costs attributable to EME were indistinguishable for vaccinated and nonvaccinated horses.