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ISSN : 2287-7991(Print)
ISSN : 2287-8009(Online)
Journal of the Preventive Veterinary Medicine Vol.37 No.1 pp.53-55
DOI :

Detection of Bartonella species from cattle ticks in South Korea

Se Eun Choe†, Ji-Yeon Kim, Myeong Ju Chae, Sung-Il Kang, Moon Her, Jeong Hui Gu, Hyang Keun Lee, Kichan Lee, Seung Won Kang, Suk Chan Jung
Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang 430-757, Republic of Korea
Received 1 February 2013, Accepted 18 March 2013

Abstract

During 2010~2011, 877 ticks (Hemaphysalis longicornis and Ixodes spp.) were collected from cattle in South Korea.According to 16S RNA PCR assay for Bartonella species, 9 of them showed specific 356-bp band products. A phylogeneticanalysis based on the 16S RNA showed two clusters of Bartonella strains. One cluster included four isolates that were quitesimilar to those previously registered on Genbank from various sources, whereas the other was highly related to the referencestrain from human. It suggested that cattle ticks harbouring various Bartonella spp. might circulate among human, animals,and farm surroundings. Therefore, as ticks are main vectors that transfer pathogens affecting human beings and domesticcattle, further studies are required for bartonellosis in cattle.

11 Short Communication 최은서_가편집(1)_1교+가제본.pdf758.0KB

 Bartonellae is a Gram-negative, rod-shaped, and facultative intracellular pathogen, which can be transferred to susceptible hosts via blood-sucking arthropods, including ticks, lice, flea, and mites [2, 7]. To date, over 26 species and sub-species of Bartonella have been officially investigated [5]. In Korea, some case reports and research articles on Bartonella henselae infection in human patients and companion animals have been previously published [3, 6]. B. elizabethae infection has also been reported in rodents and small mammals [3]. Ixodes species ticks are reported to be related with Bartonella infection. Haemaphysalis longicornis is the most dominant tick species in South Korea [3]. To our knowledge, no study has reported the presence of Bartonella species in cattle in South Korea. We report the presence of Bartonella species in cattle ticks and provide the genetic relationship between these species and Bartonella isolates registered in GenBank.

 During 2010~2011, ticks were removed from cattle from five different provinces of Korea (Gyeongbuk, Chungbuk, Jeonbuk, Jeonnam and Jeju) in South Korea by using tick twister (Tick Twister® , France). Ticks collected from the same cattle were kept in the same container. Microscopic observation of all 877 ticks collected allowed for species-level classification and pooling into 556 samples. They were also segregated according to their developmental stage and gender and were subsequently stored in 70% ethanol.

Genomic DNA was extracted from the ticks by using the QIAamp DNA extraction kit (Qiagen, Germany) as per the kit instructions. The extracted DNAs (50 ㎕) were stored at -20℃ until use. To detect DNAs of Bartonella species, a PCR assay was performed using the BTNi-F and BTNi-R primer set, which targets the 16S RNA gene [3] and included the reference strain B. henselae (ATCC 49882). The PCR products were confirmed as 356-bp amplicons by 1.5% agarose gel electrophoresis and were visualised under UV light. The amplified products were purified using the QIAquick Gel Extraction Kit (Qiagen, Germany), cloned in a pTOP TA V2 vector (Enzynomics, Korea), and enzymatically digested and sequenced (Cosmo Genetech, Korea). Phylogenetic analysis was accomplished using the MEGA 3.1 software [4].

 From the 877 ticks, 874 ticks were identified as H. longicornis (99.66%), and three (0.34%) were identified as Ixodes species. H. longicornis was the most dominant species in grazing cattle in South Korea (Table 1). All the collected ticks were in the adult stage and included 814 females (92.8%) and 63 males (7.2%). They were mainly collected from the southern part of South Korea, that is, from Jeju Island (44.5%) and the Jeonnam province (28.3%).

A 16S RNA PCR assay for Bartonella species yielded a specific 356-bp amplicon for nine of the 556 pooled DNA samples. All samples corresponded to H. longicornis obtained from dairy cattle at the same farm located in the Jeonnam province (Table 1).

Table 1. Species of cattle ticks classified on the basis of microscopic observation and PCR results

 These nine samples were sequenced and a Basic Local Alignment Search Tool (BLAST) search was performed, which indicated that four DNA samples (JN 1106-36-1, JN 1106-45-2, JN 1106-46-2, and JN 1106-50-4) showed 97% sequence similarity to B. elizabethae (AB 246801.2 and AB 246807.1). Another four DNA samples (JN 1106-32-3, JN 1106-38-4, JN 1106-49-1, and JN 1106-135-5) showed 96% similarity to B. elizabethae sequences with the same accession numbers. Only JN 1106-31-4 sample showed high similarity (97%) to B. grahamii (AB 426636.1 and AB 426635.1). Five DNA samples (32-3, 36-4, 41-4, 49-1, and 135-5) also showed 97% similarity to the reference strain B. henselae (BX 897699.1).

 A phylogenetic tree based on the partial sequencing of 16S RNA was constructed using neighbour-joining analysis with MEGA 3.1 and showed two clusters of Bartonella strains (Fig. 1). Four samples (36-1, 45-2, 46-2, and 50-4) belonged to cluster A, which included various Bartonella species. The five samples (32-3, 36-4, 41-4, 49-1, and 135-5) were highly related to the reference strain B. henselae (ATCC 49882).

Fig. 1. Phylogenetic tree based on 356 bp of the 16S RNA partial region of Bartonella strains obtained in this study, constructed by using the MEGA 3.1 software. The tree was constructed using the neighbour-joining method with the Kimura- 2 parameter model, and the bootstrap was determined using 1,000 bootstrap data sets.

 Although the exact pathway of transmission of Bartonella species from ticks to cattle is unknown, ticks are considered as one of main vectors of bartonellosis [2]. In South Korea, Bartonella species have been previously detected in ticks, rodents, and small mammals [1, 3]. Several studies on vector-borne diseases such as ticks and fleas have already been published in neighbouring Asian countries such as China and Taiwan [4, 7]. Studying Bartonella species in cattle ticks is important for estimating economic loss in the cattle industry due to global warming- induced climate change.

 This is the first study reporting the presence of Bartonella species in cattle ticks (H. longicornis) from South Korea. The farm in Jeonnam in this study showed the prevalence of Bartonella species as well as other vectorborne pathogens such as Ehrlichia, Anaplasma, Babesia, and Theileria species; furthermore, their prevalence was higher on this farm than on other farms surveyed in this study (data not shown). Although Bartonella species were detected from only one dairy cattle farm, the current results suggest that cattle may develop bartonellosis because of ticks.

 Though the Bartonella species in this study were from the same farm, the phylogenetic tree analysis showed 2 clusters. Cluster A included four isolates that were very similar to those previously registered on GenBank and were obtained from various sources, including wild rodents; wild rabbit; deer; cattle; cats; composts; and human patients. Furthermore, the cluster B isolates were almost alike with B. henselae (ATCC 49882) from a human patient. Therefore, we can infer that H. longicornis, harbouring various Bartonella species, might circulate among human beings (farm owner and workers); cattle; rodents; nest; and composts near the farm. Furthermore, this study shows the presence of more than two species of Bartonella in domestic cattle ticks from South Korea.

 Because ticks are one of the main vectors that transfer pathogens affecting human beings and domestic cattle, further studies are necessary for investigating epidemiological relatedness between ticks and these pathogens and surveying bartonellosis in cattle.

ACKNOWLEDGEMENTS

 This study was financially supported by the Animal, Plant and Fisheries Quarantine and Inspection Agency, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea (Project No. C-AD13-2010-11-01).

Reference

1.Chae JS, Yu DH, Shringi S, Klein TA, Kim HC, Chong ST, Lee IY, Foley J. Microbial pathogens in ticks, rodents and a shrew in northern Gyeonggi-do near the DMZ, Korea. J Vet Sci. 2008, 9(3):285-293.
2.Cherry NA, Maggi RG, Cannedy AL, Breitschwerdt EB. PCR detection of Bartonella bovis and Bartonella henselae in the blood of beef cattle. Vet Microbiol. 2009, 135(3-4):308-312.
3.Kim CM, Kim JY, Yi YH, Lee MJ, Cho MR, Shah DH, Klein TA, Kim HC, Song JW, Chong ST, O'Guinn ML, Lee JS, Lee IY, Park JH, Chae JS. Detection of Bartonella species from ticks, mites and small mammals in Korea. J Vet Sci. 2005, 6(4):327-334.
4.Li DM, Liu QY, Yu DZ, Zhang JZ, Gong ZD, Song XP. Phylogenetic analysis of Bartonella detected in rodent fleas in Yunnan, China. J Wildl Dis. 2007, 43(4):609-617.
5.Mediannikov O, Davoust B, Cabre O, Rolain JM, Raoult D. Bartonellae in animals and vectors in New Caledonia. Comp Immunol Microbiol Infect Dis. 2011, 34(6):497- 501.
6.Suh B, Chun JK, Yong D, Lee YS, Jeong SH, Yang WI, Kim DS. A report of cat scratch disease in Korea confirmed by PCR amplification of the 16S-23S rRNA intergenic region of Bartonella henselae. Korean J Lab Med. 2010, 30(1):34-37.
7.Tsai YL, Chomel BB, Chang CC, Kass PH, Conrad PA, Chuang ST. Bartonella and Babesia infections in cattle and their ticks in Taiwan. Comp Immunol Microbiol Infect Dis. 2011, 34(2):179-187.