VECTOR-BORNE DISEASES,SURVEILLANCE,PREVENTION Molecular and Parasitological Survey of Bovine Piroplasms in the Black Sea Region, Including the First Report of Babesiosis Associated with Babesia divergens in Turkey M. AKTAS 1 AND S. OZUBEK Department of Parasitology, Faculty of Veterinary Medicine, University of Firat, 23119, Elazig, Turkey. J. Med. Entomol. 52(6): 1344–1350 (2015); DOI: 10.1093/jme/tjv126 ABSTRACT Clinical cases of babesiosis were evaluated, and the frequency of bovine Babesia and Theileria parasites was determined in cattle. Blood samples and thin blood smears were collected from 23 cattle exhibiting clinical signs of babesiosis. In addition, tick and blood samples were collected from 100 apparently healthy cattle cograzing from the same area. Egg masses obtained from fully engorged female ticks were included. DNA isolated from blood and tick samples was screened for Babesia and Theileria by reverse line blot assay. Piroplasms compatible with Babesia spp. were observed microscopi- cally for symptomatic cattle as circular, oval, elongated, or pear-shaped bodies. Parasitemia ranged from 0.08 to 0.9% for Babesia bovis, 2.5 to 15.4% for Babesia bigemina, and 7.4% for Babesia divergens. Reverse line blot showed positivity in 13 (13%) of the sampled clinically normal cattle and revealed the presence of three Babesia species. Babesia bovis was the most prevalent (9/100, 9%), followed by Babesia occultans (3/100, 3%) and B. bigemina (1/100, 1%). One animal infected with B. bigemina was also in- fected with B. bovis. The single animal infected with B. divergens showed symptoms of babesiosis. Ticks were identified as Rhipicephalus annulatus, Rhipicephalus turanicus, and Ixodes ricinus. One female R. annulatus and its egg mass were infected with B. bigemina. Neither Theileria annulata nor Theileria buffeli/orientalis infections were observed in cattle or ticks. This is the first report of clinical babesiosis caused by B. divergens in cattle from Turkey. KEY WORDS Babesia, PCR, reverse line blot, cattle Babesiosis, caused by intraerythrocytic apicomplexan parasites of the genus Babesia that infect a wide range of domestic and wild animals, is gaining increased in- terest as an emerging zoonosis (Penzhorn 2006). The disease is characterized by high fever, jaundice, anemia, hemoglobinuria, ataxia, anorexia, general circulatory shock, and occasionally neurological signs as a result of sequestration of infected erythrocytes in cerebral capillaries (Bock et al. 2004). Bovine babesiosis is caused by Babesia bovis, Babesia bigemina, Babesia divergens, Babesia major , and Babesia occultans. Among these, B. bovis, B. bigemina, and B. divergens are clinically important agents in the etiology of bovine babesiosis in tropical and subtropical areas (Bock et al. 2004). B. divergens is particularly significant due to its zoonotic potential (Zintl et al. 2003). Babesia spp. are transmitted by ixodid ticks of the genera Rhipicephalus, Hyalomma, Dermacentor , Haemaphysalis, and Ixodes (Bock et al. 2004, Aktas et al. 2014). The traditional method of laboratory diagnosis of piro- plasm infections is based on the microscopic detection of parasites in Giemsa-stained blood smears prepared from animals with clinical symptoms. However, this method is not sufficiently specific or sensitive to detect Babesia and Theileria parasites, particularly when mixed infections occur. Recently, PCR-based reverse line blot (RLB) hybridization has been used for the detection and identification of Babesia and Theileria species (Gub- bels et al. 1999, Georges et al. 2001, Garcia-Sanmartin et al. 2006, Ica et al. 2007b, Altay et al. 2008, 2012). The main tick-borne hemoparasitic disease occurring in cattle throughout Turkey is theileriosis (Aktas et al. 2001a, b, Dumanli et al. 2005, Inci et al. 2013). How- ever, bovine babesiosis caused by B. bovis, B. bigemina, B. major , and B. occultans have also been detected by molecular methods in some areas of the country (Duzlu et al. 2011, Yavuz et al. 2011). B. divergens has been detected by serological tests in Turkey (Dinc ¸er et al. 1991, Aktas et al. 2001a, b). We performed a PCR-based RLB survey on symptomatic and asymp- tomatic cattle in Bartın Province of the West Black Sea region of Turkey. The aims of the study were to de- scribe clinical cases of babesiosis, and to determine the frequency of bovine Babesia and Theileria parasites. Materials and Methods Study Area and Collection of Blood and Tick Samples. The study was conducted in cattle of Bartın Province in the West Black Sea region of Turkey 1 Corresponding author, e-mail: maktas@firat.edu.tr. V C The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com Downloaded from https://academic.oup.com/jme/article/52/6/1344/868705 by guest on 12 June 2022