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The aim of the present study was to detect and characterise the species and subspecies of Babesia spp. that cause canine babesiosis in Croatia. Twenty-eight dogs with typical signs of babesiosis (lethargy, anorexia, fever, dark urine and thrombocytopenia) were included in this study. Their blood smears showed the presence of Babesia canis . The results showed the detection of one subspecies, namely Babesia canis canis using PCR, and subsequent sequence analysis demonstrated portions of the nss rRNA gene in 27 out of 28 samples. Sequence analysis of the isolates showed 100% identity in 11 samples, 99.7% identity (one nucleotide difference) in 11 samples and 99.4% identity (two nucleotides difference) in 5 samples with B. canis canis . The results of this study confirm the presence of B. canis canis in infected dogs in Croatia and demonstrate a slightly new genetic variant of Babesia subspecies.

Changes in coagulation parameters were studied in dogs naturally infected with Babesia canis canis (n = 30), and haemostasis was evaluated and compared to values obtained from healthy dogs (n = 29). To date, there have not been any studies examining the dynamics of thrombin-antithrombin complex formation in cases of canine babesiosis. Coagulation parameters evaluated before (day 0) and on days 1, 2, and 3 after treatment with imidocarb (6 mg/kg inj. s.c.) included the determination of platelet counts, the formation of thrombin-antithrombin complexes (TAT), prothrombin time (PT), activated partial thromboplastin time (APTT) and antithrombin III (AT III) activity. TAT complexes were significantly elevated in animals with babesiosis on days 0 and 2 (mean 49.7 and 87.7 microg/L vs. control, 7.2 microg/L). AT III activity was significantly decreased at all time-points examined. There were no differences in PT. On days 2 and 3 the APTT was significantly shortened in the infected dogs when compared to control animals (means of 21.3 and 19.2 s vs. control, 30.0 s). Our analysis demonstrated that infected dogs had significant thrombocytopenia during the course of the study (mean day 0 - 29 x 10(9) /L, day 1 - 48 x 10(9) /L, day 2 - 47 x 10(9) /L and day 3 - 87 x 10(9) /L, vs. control -259 x 10(9) /L). These data suggest that babesiosis in dogs compromise primary and secondary haemostasis and that induction of disseminated intravascular coagulation (DIC) occurs in canine babesiosis.