J Vet Sci.  2008 Jun;9(2):155-160. 10.4142/jvs.2008.9.2.155.

Antigenic diversity of Theileria major piroplasm surface protein gene in Jeju black cattle

• Ko MS
• Lee KK
• Hwang KK
• Kim BS
• Choi GC
• Yun YM

• KMID: 1106230
• DOI: http://doi.org/10.4142/jvs.2008.9.2.155

Abstract

Piroplasms are tick-transmitted, intracellular, hemoprotozoan parasites that cause anorexia, fever, anemia, and icterus. Theileriosis is caused by Theileria sergenti and causes major economic losses in grazing cattle in Japan and Korea. In May 2003, we examined the antigenic diversity of the major piroplasm surface protein (MPSP) gene in 35 healthy Jeju black cattle that were born and raised at the National Institute of Subtropical Agriculture. On microscopic examination of Giemsa-stained blood smears, 9 of 35 cattle had intra-erythrocytic piroplasms. Hematological data were within normal range for all 35 cattle. Amplification of DNA from all blood samples using universal MPSP gene primers showed mixed infections with C, I, and B type Theileria spp. Type C was identified in 20 of 35 blood samples, and type B was identified in 17 samples. Allelic variation was seen in type B.

Keyword

cattle; MPSP gene; piroplasma; surface protein; theileriosis

MeSH Terms

Animals
Antigens, Protozoan/*genetics
Base Sequence
Cattle
DNA Primers/genetics
*Genetic Variation
Korea
Molecular Sequence Data
*Phylogeny
Protozoan Proteins/*genetics
Sequence Analysis, DNA
Theileria/*genetics
Theileriasis/*parasitology

Figure

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  Fig. 1 Restriction pattern of the PCR product amplified with primers Ts-B and Ts-R. The PCR product was digested with restriction enzymes BglI (Bg), DraI (D), EcoT14I (E14), EcoRV (EV), and HindIII (H), electrophoresed on a 2.0% agarose gel, and stained with ethidium bromide. Lane M: marker (HaeIII digested ØX174), U (undigested): PCR product. A: B-type pattern; B, C: B-type similar pattern.

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  Fig. 2 Comparison of the partial nucleotide sequences of the PCR product from Jeju black cattle (JBC-1, 2) and MPSP genes of other Theileria spp. from the GenBank database. Gaps (-) indicate spaces introduced into the aligned sequences by the multiple alignment program in CLUSTAL W. An asterisk represents identical nucleotides. T. sergenti-D50304 (Aomori), D11046 (Ikeda), AB016280 (Fukushima); T. buffeli-D11047 (Warwick); T. orientalis-AB008369 (Essex); Jeju black cattle (JBC)-1, 2 - Theileria isolate from Jeju black cattle.

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  Fig. 3 Phylogenetic tree for the MPSP gene of Theileria parasites. This phylogenetic tree was constructed using the DNASTAR program, with B. equi as an out-group. The GenBank accession numbers for the sequences used in the analysis are as follows: T. sergenti-D50304 (Aomori), D11046 (Ikeda), AB016280 (Fukushima); T. buffeli-D11047 (Warwick); T. orientalis-AB008369 (Essex); B. equi-L13784; JBC-1, 2 - Theileria isolate from Jeju black cattle.

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