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Molecular Prevalence of Canine Hepatozoonosis in Owned-Dogs in Central Part of Turkey
Erol U., Altay K., Atas A.D.,...
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Abstract
Canine hepatozoonosis is a tick-borne disease caused by Hepotozoon canis and Hepotozoon americanum. H. canis is widespread almost all over the world, while H. americanum exists only in the continental of America. The aim of this study was to determine the prevalence of canine hepatozoonosis from blood samples of 150 owned-dogs in central part of Turkey using polymerase chain reaction (PCR). Sixty seven out of 150 (44.67%) samples were PCR positive for Hepatozoon spp. Five out of 67 positive PCR products were sequenced to determine Hepatozoon species. Partial nucleotide sequences of 18S small subunit ribosomal RNA gene were compared to Hepatozoon spp. sequences registered in GenBank. Nucleotide sequencing resulted in three samples of the Hepatozoon spp.; two samples were determined to be H. canis. Five nucleotide sequences detected in this study were deposited in GenBank under accession numbers MW350127, MW350128 (Hepatozoon canis), MW350129- MW350131 (Hepatozoon spp.). In this study, canine hepatozoonosis infection rate was found to be very high (44.67%), resulting in a high rate of infection among the owneddog populations in the study areas. To the best of our knowledge, it is the first study on the etiology and epidemiology canine hepatozoonosis in Sivas province in Turkey.
Keywords: Canine Hepatozoonosis; Dog; PCR; DNA Sequences; Sivas; Turkey
Introduction
Species of the Hepatozoon genus are apicomplexan parasites belonging to the Hepatozoidae family. This genus has more than 340 Hepatozoon species. Hepatozoon species infect both domestic and wild animals such as carnivores, reptiles, amphibians, and birds (1, 2). Two species (Hepatozoon canis and Hepotozoon americanum) of the Hepatozoon genus infect domestic and wild canid species causing the disease referred to as canine hepatozoonosis (1, 3).
Hepatozoon canis infection in dogs was first reported from India in 1905. Thereafter this species has been described from almost all parts of the world (4). Other species of canine hepatozoonosis, H. americanum was detected in the U.S.A. in 1978. H. americanum was thought of as a more virulent form of H. canis at that time, but studies revealed that this was a different type of Hepatozoon species and named H. americanum in 1997 (3).
Hepatozoon canis and H. americanum are transmitted to the dogs by hard (Ixodidae) ticks. The main vector of H. canis is Rhipicephalus sanguineus (brown dog tick). H. canis is also transmitted by the tick R. turanicus and Amblyomma ovale (3, 5). H. americanum is transmitted by A. maculatum (Gulf Coast tick) in the U.S.A. (6). Most of tick-borne protozoan parasites are transmitted to their host during blood feeding. On the contrary to other tick-borne protozoan parasites, H. canis and H. americanum are transmitted to the host by ingestion of ticks or tick parts contaminated with sporozoites (1, 2, 3, 7).
Microscopic, histopathological, serological, and molecular techniques have been used to diagnose canine hepatozoonosis. Microscopic examination is mostly based upon the presence of intracellular ellipsoidal-shaped gamonts within neutrophils and rarely in monocytes in Giemsa- or Wright’s-stained blood smears (3, 4, 8). Microscopic examination may be used for identification of H. canis infection, but since H. americanum gamonts are rarely seen in blood smears, histopathological examination especially immunohistochemical techniques are more sensitive than microscopic technique for the identification of H. americanum (2, 3). The histopathology of skeletal muscle of dogs infected with H. americanum can be seen in pyogranulomatous myositis and large round or oval “onion skin” cysts containing a central nucleus (2, 3, 4, 7). ELISA and IFAT have been commonly used to diagnose canine hepatozoonosis in large epidemiological studies to determine chronic infections (2, 9, 10).
Molecular identification techniques such as PCR, RealTime PCR, and DNA sequencing have been preferred for the diagnosis of canine hepatozoonosis (11, 12). These methods have been accepted to be more sensitive and specific than the other methods for the diagnosis of canine hepatozoonosis. Furthermore, molecular methods can detect a small number of pathogen DNA in suspect material (2, 3, 4, 11, 12). Recently, some new genotypes of Hepatozoon spp. have been found in dogs using these methods (13).
Canine hepatozoonosis has been described in almost all parts of the world, such as southern Europe, the Middle East, Africa, Southeast Asia, and North and South America (2, 3, 4). The disease has also been detected in different provinces of Turkey (10, 13, 18). To date, to the best knowledge of the authors canine hepatozoonosis has not been investigated in Sivas province of Turkey. The aim of this study was to determine and to identify the presence of Hepatozoon species among owned-dogs’ population in Sivas province by using PCR and sequencing. [...]
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