• Korean Journal of Veterinary Service
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• v.34 no.3
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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.

• Korean Journal of Veterinary Research
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• v.36 no.1
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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.

• Korean Journal of Veterinary Research
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• v.35 no.1
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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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• v.30 no.3
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• pp.201-208
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• 1992

A Theileria sergenti soluble merozoite preparation containing the 29, 34, 35 and 105 KD as the immunodominant polypeptides, was evaluated for efficacy, safety and protectivity in Holstein calves against virulent field tick challenge. The soluble antigens (100 mg/dose) were fortified with either complete or incomplete Freund's adjuvant. Twenty naive calves, aged one month, were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later. Twenty additional calves served as controls. Five weeks after the booster dose, vaccinates and uninoculated controls were moved to a pasture, a heavily tick infested area in Cheju-do, Korea. The vaccinates showed negligible change in hematocrit and total RBC count whereas control animals showed significant (p<0.05) hematological changes and associated anemia. Only 30% of vaccinates required chemotherapy after the experiment was terminated. All control animals required chemotherapy and 25% received blood transfusion. The highest percent parasitized erythrocytes in vaccinated cattle was 0.4% as compared with 3.6% among controls during the month of July. A significant difference (p<0.05) was observed in the rate of body weight increase. Significant difFerences were also noted in serum albumin, aspartate aminotransferase, total protein and bilirubin. Significantly more vaccinated cattle maintained normal ranges of hematological and biochemical values as compared with the control group. It is suggested that soluble merozoite T. sergenti antigens may be potential vaccine candidates for developing a genetic vaccine in Korea.

• Korean Journal of Veterinary Research
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• v.33 no.1
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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.

• Korean Journal of Veterinary Research
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• v.39 no.4
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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.

• Journal of Veterinary Clinics
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• v.27 no.5
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• pp.501-507
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• 2010

The aim of the current study was to analyze the diversity of the major surface protein (Msp) gene in Theileria buffeli, which is known as the major antigenic protein recognized by the immune system of the host. In addition, we characterized the diversification of the Msp gene and its relationship to with the pathogenicity of Theileria. Complete blood counts (CBC) and Theileria 18S rRNA PCR sequence analysis were performed for 177 Korean indigenous cattle (KIC) in Jeju Island. A total of 28 KIC (16 anemic and 12 non-anemic KIC) were then randomly selected based on 18s rRNA PCR positive samples for sequence analysis of the Theileria Msp gene, which was performed twice for each specimen. The resulting 56 Msp gene sequences were classified into five antigenicity types (type I to V), according to the variable region (517-571 bp), which exhibited high similarity (${\geq}$ 98.9%) to several available GenBank sequences (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). The 56 Msp sequences consisted of 22, 15, 9, 8, and 2 cases of type I to type V Msp genes, respectively. The most prevalent type in both anemic and non-anemic KIC was type I (37.5% in anemic and 41.7% in non-anemic). Among the remaining types, type II was the most prevalent (37.5%) in anemic KIC, while type IV was the most prevalent (25%) in non-anemic KIC. The results of our study help confirm the diversity of Msp gene types and demonstrate that the gene type distribution of Msp genes varies among Theileria-infected KIC in Jeju Island.

• Korean Journal of Veterinary Service
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• v.42 no.2
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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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• v.19 no.6
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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.

• Korean Journal of Veterinary Research
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• v.39 no.4
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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.