BARTONELLA PATHOGENS IN SMALL MAMMALS (RODENTIA AND INSECTIVORA) AND THEIR FLEAS (SIPHONAPTERA) AND TICKS (IXODIDAE) IN

Lipatova Indr ė (1), Paulauskas Algimantas (1), Radzijevskaja Jana (1), Gedminas Vaclovas (2) (1)Faculty of Natural Sciences, Vytautas Magnus University, , Lithuania (2)Kaunas Tadas Ivanauskas Zoological Museum, Kaunas, Lithuania e.mail: [email protected], [email protected]

Introduction Bartonella is gram-negative bacteria that are transmitted by the arthropod vector such as ticks, fleas, flies and lice. This pathogen is emerging zoonotic agent and has been identified in domestic and wild mammals (Breitschwerdt et al. 2000). The genus Bartonella has about 30 species or subspecies of which at least 14 have been associated with a variety of human diseases (Morick et al. 2010).

The aim of this study was to detect and identify Bartonella pathogens in fleas and ticks collected from small mammals (rodentia and insectivora) in Lithuania.

Methods In our study were captured 318 rodents (including Apodemus flavicollis , Apodemus sylvaticus, Myodes glareolus, Microtus agrestis, Microtus arvalis, Microtus oeconomus and Micromys minutus ) and 48 insectivores ( Sorex minutus) with live-traps (Fig. 1) in the peninsula Curonian Spit of Lithuania (Fig. 2). One hundred eighty four fleas (identified 5 speces: Hystrichopsylla t. talpae, Ctenophthalmus a. agyrtes, Palaeopsylla soricis, Megabothris turbidus and Megabothris walkeri ) and 111 ticks ( Ixodes ricinus ) were collected from small mammals (Fig. 3). All samples were tested by polymerase chain reaction for the detection of Bartonella using BhCS.781p/BhCS.1137n primers (Dong et al. 2007). Differentiation of Bartonella species was done by nested PCR of the ITS region and the gltA gene (table 1) (Kaewmongkol 2012) . A nested PCR of the ITS region was performed using WITS-F/WITS-R and Bh311-332F/Bh473-452R primers. A nested PCR of the gltA gene was performed using gltA-F2/gltA-R2 and BhCS.781F/BhCS.1137R. PCR products were purified and subjected to sequence analysis. The obtained sequences were aligned with gene sequences of Bartonella pathogens registered in GenBank database.

Table 1. PCR conditions and primers used for the detection of Bartonella pathogens

Fig. 1. Live-traps

Results Bartonella pathogens were isolated from 27% small mammals, 30% fleas and 25% pools of ticks. A nested PCR and sequencing analysis Fig. 2. Rodents and insectivores capturing place of PCR products identified Bartonella pathogens to three species which can cause disease in human. B. grahamii was detected in 41% small mammals, 38% fleas and 11% pools of ticks; B. taylorii was detected in 12% small mammals, 35% fleas and 18% pools of ticks and 4% fleas were positive for both pathogens. Also we have detected B. rochalimae in 2% flea.

Conclusions In Lithuania only B. henselae was diagnosed for patient. Fleas have previously been involved in the transmission of B. henselae infection. Our study confirms that fleas and ticks are vector of Bartonella pathogens in small mammals. This is the first detection of B. grahamii, B. taylorii and B. rochalimae in Lithuania. B. grahamii and B. rochalimae have previously been associated with human disease. Fleas and ticks may be involved in the transmission of rodent Bartonella pathogens some of which can be pathogenic to humans.

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