• 한국동물위생학회지
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• 제34권3호
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• pp.251-258
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• 2011

This study was performed to determine the prevalence of Theileria sergenti (T. sergenti) in cattle reared in eastern areas of Gyeongbuk province by PCR. Among 443 samples collected from 42 cattle farms, 96 (21.7%) samples from 13 (31.0%) farms were positive for T. sergenti. By regions, 87 (26.6%) out of 327 cattle in Gyeongju, 8 (9.5%) out of 84 cattle in Pohang, and 1 (3.1%) out of 32 cattle in Ulleung were positive for T. sergenti. T. sergenti infection in dairy cattle (27.9%) was significantly higher than that in Korean cattle (9.4%, P<0.0001). Accordingly, Korean cattle were more resistant to T. sergenti infection. Prevalence of T. sergenti in cattle was increased with age (P<0.0001). The infection rate in cows (23.3%) was significantly higher than that in bulls (5.0%, P<0.01). Seasonally, prevalence of T. sergenti in cattle was highest in autumn (32.7%, P<0.01). Prevalence of T. sergenti in grazing cattle (66.7%) was significantly higher than that in non-grazing cattle (15.8%, P<0.0001). Since prevalence of T. sergenti infection is high in cattle reared in eastern areas of Gyeongbuk province, survey on other hemoparasites and appropriate control programs need to be established in this region.

• 대한수의학회지
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• 제36권1호
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• pp.187-193
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• 1996

The T sergenti DNA fragments used as probes of KTS1(2.4kb) and KTS3(1.5kb) were labeled with digoxigenin-11-dUTP for the Southern hybridization. T sergenti DNAs from different geographic locations(Korea; Chonbuk, Kyungbuk, Chungnam, Kangwon, Cheju island, Japan; Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9202, Shintoku 9102) which had been digested with Pst I and EcoR I were probed by the digoxigenin-11-dUTP-labeled KTS1 and KTS3. As the results, the samples from Chonbuk, Kyungbuk, Cheju island in Korea and Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9102 in Japan were positively reacted, but the others from the other locations not reacted. In the comformation test of T sergenti DNA from different geographic locations, all of the samples were positively detected by PCR amplification.

• 대한수의학회지
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• 제35권1호
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• pp.87-94
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• 1995

Western immunoblot analysis of antigen of T sergenti merozoite revealed that the immunodominant proteins of this organism were characterized as the 18KD, 29KD, 34KD, 45KD and 105KD in Korea. The 34KD and 45KD among those immunodominant proteins of the parasite were isolated and their amino acid sequences from the $NH_2$-terminus were determined and synthesized. They respective polypeptides were cationized to enhance their antigenicity, fortified with Freund's adjuvant and tested for immunogenicity in rabbits and cattle. The results obtained were as follows; 1. Theileria sergenti merozoite antigen was shown in 120KD, 100KD, 66KD, 45KD, 34KD and 30KD in western immunoblot using serum of rabbits immunized with 34KD synthetic polypeptide and 70KD, 58KD, 55KD and 45KD using bovine serum. In western immunoblot, 45KD, 34KD and 30KD were recognized by immunized rabbits, and 50KD and 45KD by cattle sera immunized with 45KD synthetic polypeptide, respectively. 2. The ELISA utilizing the synthetic polypeptides demonstrated significant antibody response to the respective peptides. After the 2nd booster injection, an OD of 0.760(preimmunization 0.132) in rabbits and an OD of 0.645(preimmunization 0.488) to 34KD synthetic polypeptide in cattle were observed. In animals immunized with 45KD synthetic polypeptide, after the 2nd booster injection, an OD of 0.640(preimmunization 0.144) in rabbit, and an OD of 0.776 (preimmunization 0.477) in cattle were measured. 3. After the 2nd booster the reciprocal IFA titer was 1:64 in rabbits and 1:512 in cattle immunized with the 34KD synthetic polypeptide. The IFA titre was observed as 1:512 in rabbit and 1:1,024 in cattle in immunized with the 45KD synthetic polypeptide.

• Parasites, Hosts and Diseases
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• 제30권3호
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• pp.201-208
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• 1992

Theileria sergenti merozoite의 면역원성 polypeptide중 29, 34, 35 그리고 105KD를 함유한, 수용성 항원을 흘스타인 송아지 (1개월령)에게 접종한 후, 제주도의 야외 목장에 방목시킴으로써 진드기로부터 병원성 충주의 도전에 따른 항원의 효력, 안전성 그리고 방어력을 관찰하였다. 즉 송아지 20두에 수용성 항원(100 mg/dose)을 Freund's adjuvant와 함께 1차 접종, 4주 후에 추가접종하였으며, 다른 20두의 송아지를 대조군으로 사용하였다. 추가접종 5주 후에 이들 송아지를 야외 목장에 방목, 매월 혈액학적 그리고 생화학적 변화를 관찰하였다 예방접종군에 있어서 hematocrit와 총적혈구수는 진드기 도전 전과 비교하여 약간의 변화를 인정할 수 있었다. 일반적인 혈액학적 소견은 예방접종군과 비교하여 유의한 차이를 인정할 수 있었다(p<0.05). 7월에 있어서 적혈구내 기생률은 예방접종군에 있어서는 0.4%이었으나, 대조군에 있어서는 약 3.6%이었다. 체중 증가율, albumin, aspartate alninotransferase, total protein, 그리고 bilirubin에 있어서도 유의한 차이 (p<0.05)를 인정할 수 있었다. 예방접종군의 혈액학적 그리고 생화학적 측정치는 대조군 보다는 정상치에 접근하였다. 실험종료 직후에 있어서 예방접종군의 30% 송아지와 대조군의 모든 송아지의 치료와 대조군 중 25%는 수혈이 요구되었다. T. sergenti merozoite의 수용성 항원에 대한 이 같은 연구 결과는 앞으로 유전공학적 백신제조를 위한 연구자료로 활용될 수 있을 것이다.

• 대한수의학회지
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• 제33권1호
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• pp.137-143
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• 1993

This study was carried out to completelycure the experimental bovine theileriasis with small unilamella vesicle liposome incorporated buparvaquone which was effective both to schizonts in lymphocyte of lymph nodes and piroplasmic stage in erythrocytes. Small unilamella vesicle liposome incorporated buparvaquone was prepared by French pressure cell method using egg phosphatidylcholine. The diameter of the vesicles was ranged from 5 to 220 nm, but the most vesicles were ranged from 10 to 50 nm in diameter. The incorporation rate for buparvaquone was 100%. Parasitaemia of the 10 calves inoculated with $5{\times}10^8$ erythrocytes infected with Theileria sergenti were first detected from on day 16 to day 23 after inoculation. In calves treated with a dose rate 2.5 mg/kg BW of free buparvaquone, a gradual decrease in piroplasmic parasitaemia was observed following treatment to day 5. However parasitaemia levels returned to near pretreatment values after approximately 60 days. In calves treated with a dose rate 5.0mg/kg BW of free buparvaquone, parasitaemia were disappeared on day 3 after treatment, but there was a mild recrudescence of infection on day 28 after treatment. In calves treated intraavenously with a dose rate 2.5 mg/kg BW of buparvaquone incorporated in liposome, the calves were all cured on day 2 or day 3 after treatment. In calves treated subcutaneously and intraperiotoneally with a dose rate 2.5 mg/kg BW of buparvaquone incorporated in liposome, parasitaemia were disappeared on day 3 or day 4 after treatment, but there was a mild recrudescence of infection on day 40 or day 45 after treatment.

• 대한수의학회지
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• 제39권4호
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• pp.786-796
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• 1999

As an attempt to develop an effective control method against theileriosis, recombinant antigen protein was produced. Thirty-two kDa membrane protein(MP) gene of T sergenti was amplified through RT-PCR from extracted total RNA of T sergenti isolated in Chonbuk, Korea. The amplified 869 bp of Korean T sergenti membrane gene was cloned and the base sequences were analyzed. The amplified gene was cloned into E coli expression vector, pQE32 plasmid vector, and the vector was introduced into E coli strain M15 to produce the recombinant membrane protein. For the induction of T sergenti membrane protein(KTs-MP), the plasmid harboring E coli strain M15 were cultured in the presence of IPTG, and the recombinant protein were purified by $Ni^+$-NTA agarose. Then, to confirm the authenticity of the produced membrane protein, molecular weight of expressed recombinant KTs-MP was analyzed by SDS-PAGE and Western blotting. The molecular weight of expressed recombinant protein was 32 kDa as expected. The recombinant KTs-MP was successfully recognized by anti-His Tag antibody, antisera of T sergenti infected cattle and monoclonal antibody of T sergenti membrane protein. Therefore, we concluded that the authentic 32 kDa membrane protein of T sergenti was produced as immunologically recognizable form.

• 한국임상수의학회지
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• 제27권5호
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• pp.501-507
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• 2010

본 연구는 국내 타일레리아에서 주요항원단백질(major surface protein) 유전자의 다양성을 분석해 보고자 수행되었다. 나아가 Msp 유전자의 다양성과 타일레리아의 병원성과의 관계도 분석하였다. 제주에 있는 목장으로부터 총 177마리의 한우 혈액을 공시재료로 사용하여 혈액검사와 18S rRNA를 표적으로 하는 PCR을 실행하였다. 그 후, 타일레리아 18S rRNA에 양성인 28마리 (16마리 빈혈군과 12마리의 정상군)를 무작위로 선발하여 Msp유전자의 염기 서열을 반복하여 분석하였다. 총 56개의 염기서열 결과는 다변성 부위(517-571 bp)에 따라 크게 type I에서 type V까지 5가지 형태로 나눌 수 있었는데, 이는 유전자은행(GenBank)에 등록되어 있는 다음의 유전자와 98.9% 이상 일치하였다 (Theileria spp. from China-EU584237; T. sergenti from China-DQ078264; Theileria spp. from Thailand-AB081329; Theileria spp. from Japan-AB218442; T. sergenti from Japan-AB016280). 그 분포는 22, 15, 9, 8, 2개가 각각 type I에서 V까지 분포하였고 빈혈과 관계없이 type I이 가장 많이 나타나는 것으로 밝혀졌다(37.5%의 빈혈군과 41.7%의 정상군). 나머지 type중에서는 type II가 빈혈군에서 가장 많이(37.5%) 나타났으며, 반면 type IV는 정상군에서 많이 (25%) 나타났다. 본 연구는 국내 타일레리아 Msp유전자의 다양성을 밝히는데 좋은 자료로 활용될 수 있을 것이다.

• 한국동물위생학회지
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• 제42권2호
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• pp.113-116
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• 2019

Bovine theileriosis caused by Theileria sergenti is a tick-borne hematoprotozoan disease that is characterized by chronic anemia and fever in cattle. In this study, results of microscopic examination and PCR detection confirmed 17 Korean native cattle with emaciation and fever as acute bovine theileriosis caused by T. sergenti. Buparvaquone was injected as treatment, but was proved to be an inappropriate measure according to our study. After 6 months of injection, clinical signs and hematological values were recovered, but T. sergenti was still identified in blood sample as a result of microscopic exam and PCR. These results suggest that continuous management is necessary to control bovine theileriosis. Therefore, findings of this study may provide significant guideline on the control of bovine theileriosis.

• Journal of Veterinary Science
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• 제19권6호
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• pp.846-849
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• 2018

A group of benign Theileria species, which are often referred to as T. orientalis/T. buffeli/T. sergenti group, has low pathogenicity in cattle. Herein, we report on Theileria spp. in cattle on a farm from China. Based on phylogenetic analysis of the major piroplasm surface protein gene sequences, we detected 6 genotypes that were categorized as Types 1, 2, 3, 4, and 5 as well as an additional Type 9 genotype. The new epidemiological features of the T. orientalis/T. buffeli/T. sergenti parasites in China indicate a greater diversity in the genetics of these species than had been previously thought.

• 대한수의학회지
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• 제39권4호
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• pp.797-805
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• 1999

T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.