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Canine Leishmaniosis Spread in Croatia: Feasibilities of PCR-Based and Serological Monitoring Activities

In the south littoral parts of Croatia canine leishmaniosis (CanL) had been recognized as a problem for the first time in the first part of 20th century. Human visceral leishmaniosis has been sporadically reported in Croatia, and as with other diseases transmitted by arthropods, it is not a major public health problem in Croatia. Since 1997 CanL in Croatia has been proven parasitologically and serologically in hundreds of symptomatic and asymptomatic dogs . Isoenzymatic analysis from 30 in vitro cultivated isolates (performed in WHO Centre for Leishmaniosis in Madrid, Spain and in Istituto Superiore di Sanità in Rome, Italy) revealed: Leishmania infantum (MON-1 ; MON-27 ; and MON -34). All the dogs visited or lived in central and south parts of Dalmatia (from the city of Trogir along the coast and city of Drnis in the hinterlands in the West, Montenegro border in the East, Bosnia and Herzegovina border in the North, the Adriatic sea in the South, as well as on middle and south Dalmatian islands). During 2005, 1776 healthy dogs sera originating from central and southern parts of Dalmatia had been serologically investigated by indirect immunofluorescence (home antigen). During the autumn in 2006, field samples taken from 200 clinically healthy dogs originating from south littoral Croatia (Dubrovnik area) were serologically tested by indirect immunofluorescence with home antigen, and a PCR-based protocol for the detection of Leishmania parasites in canine whole blood was performed. The parasite DNA had been extracted using QIAmp DNA Mini Kit® (QIAGEN). Polymerase chain reaction was performed on the highly reiterated minicircle kDNA from dogs peripheral blood that amplified a fragment of 145 bp.In 2005, 141 (7, 9%) sera among 1776 reacted positive (titre ≥ 80-1). In 2006, 16 among 200 dogs sera (8, 0 %) reacted positive with titre ≥ 80-1 and the 145 bp fragment originated from kDNA was detected in 161 among 200 whole blood samples (80, 5%). Since the PCR-based monitoring protocol revealed the infection prevalence, further serological and especially PCR monitoring activities among dogs in areas bordering to enzootic should be useful in identification of new positive canines before stable foci of CanL were developed.

dog; leishmaniosis; PCR; monitoring; Croatia

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