Concomitant Babesia gibsoni and Ehrlichia canis infection in a dog

Experimental transmission of Babesia gibsoni in a dog, in a state of subclinical infection by Ehrlichia canis, resulted in a severe concomitant infection of E. canis and B. gibsoni characterized by neurological signs, including nystagmus and seizures.Mononuclear cells harbouring the characteristic morulae of E. canis were numerous in lung impression smears.B. gibsoni were observed in the peripheral blood in erythrocytes and within macrophages in spleen, liver and kidney impression smears.Erythrocytes were found trapped within capillaries of the brain.B. gibsoni were seen in various stages of division and on average 16 merozoites were seen in a single infected erythrocyte. Key words: ehrlichiosis, babesiosis, dogVet. arhiv 75 (6), [513][514][515][516][517][518][519][520] 2005 in a dog that was in the subclinical phase of ehrlichiosis.It also focuses attention on the resultant syndrome caused by the mixed infection of E. canis and B. gibsoni. Materials and methodsAn isolate from a dog with naturally occurring canine ehrlichiosis diagnosed at the Madras Veterinary College Clinics was used.The dog had a history of tick infestation and had pyrexia and enlargement of popliteal lymph nodes.Microscopic examination of peripheral blood smears revealed the presence of Ehrlichial inclusions in mononuclear cells.Babesia gibsoni was isolated from a dog that was naturally infected with B. gibsoni, at the Madras Veterinary College Clinics.The dog was an intact, one-year-old male Labrador retriever with a history of exposure to ticks.The dog was pyrexic and had blanched conjunctival mucous membranes.Microscopic examination of peripheral blood smears revealed the presence of B. gibsoni piroplasms in RBC.The experimental dog was housed in a kennel with tick-proof conditions and was vaccinated against canine distemper, infectious canine hepatitis and leptospirosis.The dog was also treated with anthelmintic before its use in the experiment.Freedom from E. canis was confi rmed on the basis of negative results with ELISA and Dot-ELISA and freedom from haemoprotozoan infections was ascertained by thorough screening of peripheral blood smears for 10 days before being used in the experiment.Experiment I: Experimental dog aged about 6 months, seronegative for E. canis by ELISA and Dot-ELISA, was inoculated with fi ve ml of heparinized blood collected from a dog naturally infected with E. canis.Experiment II: The same dog was inoculated with 5 ml of B. gibsoni-infected blood after 90 days post-infection with canine ehrlichiosis.Sera were analyzed for E. canis antibodies by ELISA and Dot-ELISA using DH 82 cells infected with E. canis (Oklahoma strain).The sera were examined before and after experimental infection with E. canis.Antigen and known positive serum was provided by the Department of Infectious Diseases and Physiology, College of Veterinary Medicine, Stillwater, USA.The two tests were used to examine sera from suspected dogs.ELISA was carried out as per standard methods (GRAY et al., 1980).Antigen diluted 1:500 in 0.05 M carbonate/bicarbonate buffer (pH 9.6) was dispensed in volumes of 0.1 ml into wells of polystyrene microplates (Nunc, USA).After washing with PBS-T 20 three times, plate was blocked for one hour with bovine serum albumin (Merck).Test sera were diluted 1:100 in PBS and incubated with antigen for 1 h at room temperature.Bound antibody was detected using 0.1 ml of anti-dog rabbit IgG conjugated with horse radish peroxidase (Sigma, USA)

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