Phylogenetic analysis of benign Theileria species based on major piroplasm surface protein (MPSP) genes from ticks of grazing cattle in Korea

Parasitology and Insect Disease Research Laboratory, Animal, Plant and Fisheries Quarantine and Inspection Agency, 480 Anyang 6 dong, Anyang City 420-480, Republic of Korea.
Veterinary Parasitology (Impact Factor: 2.46). 05/2012; 189(2-4):145-52. DOI: 10.1016/j.vetpar.2012.04.038
Source: PubMed


Complete major piroplasm surface protein (MPSP) gene sequences of benign Theileria parasites were isolated from ticks of grazing cattle in Korea. A total of 556 tick samples were collected in five provinces: Chungbuk, Jeonbuk, Jeonnam, Gyeongbuk, and Jeju during 2010-2011. Fifteen samples from Chungbuk and Jeonnam were positive for the Theileria MPSP gene by PCR amplification using a specific primer set. A phylogenetic tree was constructed with the amplified gene sequences and 26 additional sequences published in GenBank. The benign Theileria parasites were classified into eight types, those isolated from Korean cattle ticks belonged to Types 1 (Ikeda), 2 (Chitose), 4, and 8. Types 2 and 4 were the most common types, with the rate of 40%, followed by Types 1 and 8 (with the rate of 13% and 7%, respectively). Nucleotide sequence identities of 23 theilerial MPSP sequences (15 MPSP gene sequences amplified and 8 sequences published) ranged from 67.3 to 99.8%. Multiple alignments of the deduced amino acid sequences also showed that each type was characterized by specific amino acids: 7 for Type 1, 9 for Type 2, 4 for Type 4, and 3 for Type 8.

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    • "Other investigators showed substantially less sequence variation in the widely-used mpsp gene within genotypes (buffeli: 0–5.9%; chitose: 0–2%; ikeda: 0–1.7%) compared with differences among them (11.9–19.4%) (Kim et al., 1998; Kawazu et al., 1999; Gubbels et al., 2000; Zakimi et al., 2006; Ota et al., 2009; Altangerel et al., 2011; Islam et al., 2011; Kamau et al., 2011a; Khukhuu et al., 2011; Cufos et al., 2012; Kang et al., 2012; Perera et al., 2013; Sivakumar et al., 2013). Thus, although inter-genotypic differences determined in the present study are lower than those obtained in previous studies for other nuclear genes, they are substantially higher than the degree of intra-genotypic sequence variability, which indicates that the p23 gene region employed here is useful for the identification of T. orientalis genotypes in Victoria, Australia, and is likely applicable in other parts of the world. "
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    ABSTRACT: Abstract Oriental theileriosis is a tick-borne, protozoan disease of cattle caused by one or more genotypes of Theileria orientalis complex. In this study, we assessed sequence variability in a region of the 23 kDa piroplasm membrane protein (p23) gene within and among three T. orientalis genotypes (designated buffeli, chitose and ikeda) in south-eastern Australia. Genomic DNA (n = 100) was extracted from blood of infected cattle from various locations endemic for oriental theileriosis and tested by polymerase chain reaction (PCR)-coupled mutation scanning (single-strand conformation polymorphism (SSCP)) and targeted sequencing analysis. Eight distinct sequences represented all DNA samples, and three genotypes were found: buffeli (n = 3), chitose (3) and ikeda (2). Nucleotide pairwise comparisons among these eight sequences revealed considerably higher variability among the genotypes (6.6–11.7%) than within them (0–1.9%), indicating that the p23 gene region allows the accurate identification of T. orientalis genotypes. In the future, we will combine this gene with other molecular markers to study the genetic structure of T. orientalis populations in Australasia, which will pave the way to establish a highly sensitive and specific PCR-based assay for genotypic diagnosis of infection and for assessing levels of parasitaemia in cattle.
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    • "Each MPSP sequence determined was compared with reference sequences available for members of the T. orientalis complex in current databases (15 January 2013), and sequence identities (in %) calculated by pairwise comparison . Subsequently, all MPSP sequences were aligned with reference sequences of T. orientalis, and of T. annulata and T. parva (outgroups) (Altangerel et al. (2011), Govaerts et al. (1998), Kang et al. (2012), Kawazu et al. (1992, 1999), Kim et al. (1998), Khukhuu et al. (2011), Matsuba et al. (1993), Sarataphan et al. (2003), Shiels et al. (1995), Sivakumar et al. (2012); see Supplementary Table 1) over a consensus length of 309 bp. The phylogenetic analysis of sequence data was conducted using Bayesian inference (BI) and Neighbour-Joining methods (NJ) (cf. "
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    ABSTRACT: This study investigated Theileria orientalis following outbreaks of oriental theileriosis in cattle in the state of Victoria, Australia, from September 2010 to January 2012, using traditional and molecular methods of diagnosis. A questionnaire was used to collect epidemiological information from cattle farms. Blood samples (n=301), collected from individual symptomatic and asymptomatic cattle from 19 cattle farms, were examined for the presence of Theileria on stained blood smears and tested using a PCR-based approach, employing a region within the major piroplasm surface protein (MPSP) gene as a marker. The microscopic examination of stained blood smears detected stages consistent with Theileria piroplasms in 28.1% (79/281) of the samples. PCR products were amplified from 70.8% (213/301) of the samples. Mutation scanning analysis of all amplicons displayed seven distinct profiles. Following the direct sequencing of representative amplicons, the genotypes ikeda, chitose, buffeli and type 5 were detected in 91.1%, 32.9%, 2.4% and 1.4% of 213 blood samples, respectively. The distribution of these four genotypes varied among the 19 farms; genotype ikeda was detected on all farms, whereas genotypes chitose, buffeli and type 5 were detected on 14, 3 and 2 farms, respectively. Mix infections with genotypes ikeda and chitose were common (21.6%). Survey results revealed that oriental theileriosis affected mainly beef cows of more than two years of age, prior to calving, and disease was associated with abortion and cow deaths. Future investigations should focus on developing improved tools for investigating and managing oriental theileriosis.
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