Rapid Diagnosis of Babesia gibsoni by Point-of-Need Testing by Insulated Isothermal PCR in Dogs at High Risk of Infection.

BACKGROUND: Dogs seized by law enforcement agencies during dogfighting investigations are at increased risk of Babesia gibsoni infection. A rapid and cost-effective diagnostic test would increase the feasibility of mass screening of dogs for infection and monitoring treatment efficacy in B. gibsoni-infected dogs. OBJECTIVE: To determine the performance of a point-of-need insulated isothermal PCR (iiPCR) test for diagnosis of B. gibsoni in dogs rescued in dogfighting investigations. ANIMALS: Two hundred and thirty-three dogs seized in dogfighting investigations. METHODS: Cross-sectional study. Whole blood samples were tested for B. gibsoni and Babesia spp. by iiPCR. Results were compared to a reference standard comprised of concordant results from real-time PCR in a commercial diagnostic laboratory and antibody titers. RESULTS: The iiPCR system was quick to learn, portable, and had a short processing time of <2 hours. Sensitivity and specificity of the iiPCR assay for B. gibsoni were 90% (95% confidence interval [CI] 81-95%) and 99% (CI, 95-100%), respectively. Sensitivity and specificity of the iiPCR assay for Babesia spp. were 87% (CI, 78-93%) and 98% (CI, 0.94-99%), respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: The iiPCR system produced few false-positive results, indicating that positive results are likely to represent true infections when used in high-risk animals. The iiPCR system can fail to identify 10-15% of truly infected dogs. However, the portability, speed, and economy of the iiPCR system compared to testing through a reference laboratory can allow rescue groups to screen and identify infection in more dogs.

Efficacy of Azithromycin and Compounded Atovaquone for Treatment of Babesia gibsoni in Dogs.

BACKGROUND: Approximately one-third of dogs confiscated during dogfighting investigations are infected with Babesia gibsoni. Traditional management of B. gibsoni with polymerase chain reaction (PCR)-screening, treatment with commercially available azithromycin and atovaquone, and PCR testing after 60 and 90 days is costly and impractical for large numbers of dogs at a time. HYPOTHESIS/OBJECTIVES: To assess the efficacy of an alternative protocol in which commercial atovaquone was replaced by compounded medication and PCR monitoring was initiated at 30 days after the end of treatment to decrease the total management time. METHODS: Prospective observational study. Forty-two pit bull-type dogs confiscated as part of an investigation of dogfighting, diagnosed with B. gibsoni infection, and judged to be suitable for adoption were treated with azithromycin (10 mg/kg PO q24h) and compounded atovaquone (13.4 mg/kg PO q8h with a fatty meal) for 10 days. PCR testing was repeated at 30 and 60 days after end of treatment if dogs with positive PCR tests at either time were tested at 90 days. Treatment was considered successful; 2 PCR tests 30 days apart were negative. RESULTS: Treatment was successful in 39 dogs (93%) as defined by 2 consecutive PCR-negative test results 30 days apart. In 38 dogs (90%), PCR results were the same at 30 and 60 days. CONCLUSIONS AND CLINICAL IMPORTANCE: Use of compounded atovaquone and a reduced monitoring period can reduce costs and holding times without compromising treatment efficacy. This more economical protocol can remove barriers to mass screening and management of B. gibsoni infections in dogfighting cases.

Infectious diseases in dogs rescued during dogfighting investigations.

Dogs used for dogfighting often receive minimal preventive health care, and the potential for spread of infectious diseases is high. The purpose of this study was to describe the prevalence of infectious diseases in dogs rescued from fighting operations to guide medical protocols for their immediate and long-term care. A total of 269 pit bull-type dogs were seized in a multi-state investigation. Fleas were present on most dogs, but few ticks were observed. Testing performed at intake included packed cell volume (PCV), serology and PCR for vector-borne pathogens, and fecal analysis. The most common infections were Babesia gibsoni (39%), 'Candidatus Mycoplasma haematoparvum' (32%), Mycoplasma haemocanis (30%), Dirofilaria immitis (12%), and Ancylostoma (23%). Anemia was associated with B. gibsoni infection (63% of infected dogs, odds ratio = 2.5, P <0.001), but not with hemotropic mycoplasmas or Ancylostoma. Pit bull heritage and dogfighting are known risk factors for B. gibsoni infection, possibly via blood transmission from bites and vertical transmission. Hemotropic mycoplasmas have a similar risk pattern. Empirical care for dogs from dogfighting cases should include broad-spectrum internal and external parasiticides and monitoring for anemia. Dogfighting case responders should be prepared for mass screening and treatment of B. gibsoni and heartworm infections and should implement protocols to prevent transmission of infectious and zoonotic diseases in the shelter and following adoption. Former fighting dogs and dogs with possible dog bite scars should not be used as blood donors due to the risk of vector-borne pathogens that can escape detection and for which curative treatment is difficult to document.

The effects of processing and low dose irradiation on cortical bone grafts.

The authors studied the effects of standard processing and preprocessing low dose gamma irradiation (1.5 Mrad) on the strength and incorporation of syngeneic and allogeneic cortical bone grafts. Bilateral femoral middiaphyseal 8-mm segmental defects in 120 male Fisher rats were stabilized with internal fixation. Each defect received one of six types of grafts: fresh syngeneic, processed syngeneic, irradiated processed syngeneic, fresh allogeneic, processed allogeneic, and irradiated processed allogeneic grafts. Graft processing included soaking in 70% ethanol and deep freezing for preservation. Irradiation was performed by 60Co source immediately before processing. Grafts were evaluated by histologic analysis, histomorphometric analysis, and biomechanical testing at 4 and 6 months after surgery. Graft treatment, either processing or irradiation processing, did not affect consistently or significantly the incorporation of syngeneic or allogeneic grafts. Graft allogenicity was the major determinant of the revascularization and the histologic pattern of graft incorporation. Processed and irradiated processed allogeneic grafts gained compressive strength with time and were as strong as syngeneic grafts at 6 months. Biomechanical and histologic data from this study suggest that standard processing and preprocessing low dose irradiation do not compromise the natural course of allogeneic cortical bone graft incorporation.