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.
Agriculture ; Animals ; Babesiosis ; Cattle ; China ; Genetics ; Genotype ; Parasites ; Phylogeny ; Theileria ; Virulence

Tick-borne pathogens can cause serious problems in grazing cattle. However, little information is available on tick-mediated diseases in cattle grazing on mountains. Thus, this study aimed to understand the potential problems related to tick-borne diseases in grazing cattle through the investigation of prevalent tick-transmitted infections, and their associated hematological changes, in terms of season and grazing type in Korean indigenous cattle (=Hanwoo). Hanwoo cattle from 3 regions of the Republic of Korea (=Korea) were either maintained indoors or placed on grassy mountains from spring to fall of 2014 and 2015. Cattle that grazed in mountainous areas showed a greater prevalence of tick-borne infections with an increased Theileria orientalis infection rate (54.7%) compared to that in non-grazing cattle (16.3%) (P < 0.001). Accordingly, the red blood cell (RBC) count and hematocrit (HCT) values of grazing cattle were significantly lower than those of non-grazing cattle throughout the season (P < 0.05). Moreover, RBC, hemoglobin (Hb), and HCT of T. orientalis-positive group were significantly lower than those of T. orientalis-negative group (P < 0.05). T. orientalis is a widespread tick-borne pathogen in Korea. Grazing of cattle in mountainous areas is closely associated with an increase in T. orientalis infection (RR=3.4, P < 0.001), and with consequent decreases in RBC count and HCT. Thus, these findings suggest that the Hanwoo cattle in mountainous areas of Korea are at a high risk of infection by T. orientalis, which can lead to hematological alterations. This study highlights the necessity of preventive strategies that target T. orientalis infection.
Animals ; Cattle* ; Erythrocytes ; Hematocrit ; Korea ; Prevalence ; Republic of Korea ; Seasons ; Theileria* ; Tick-Borne Diseases

Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (β-actin), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.
Animals ; B-Lymphocytes/parasitology ; Cattle ; Cell Line ; Cells/*parasitology ; Cells, Cultured ; Gene Expression Profiling ; Host-Parasite Interactions/*genetics ; Real-Time Polymerase Chain Reaction/*veterinary ; Reproducibility of Results ; Signal Transduction/*genetics ; Theileria annulata/physiology ; Theileriasis/*physiopathology

The objective of this study was to investigate the prevalence of tick-borne pathogens in the Korean water deer (Hydropotes inermis argyropus). Pathogens were identified using PCR which included Anaplasma, Ehrlichia, Rickettsia, and Theileria. Rickettsia was not detected, whereas Anaplasma, Ehrlichia, and Theileria infections were detected in 4, 2, and 8 animals, respectively. The most prevalent pathogen was Theileria. Of the 8 Theileria-positive animals, 2 were mixed-infected with 3 pathogens (Anaplasma, Ehrlichia, and Theileria) and another 2 animals showed mixed-infection with 2 pathogens (Anaplasma and Theileria). Sequencing analysis was used to verify the PCR results. The pathogens found in this study were identified as Anaplasma phagocytophilum, Ehrlichia canis, and Theileria sp. To the best of our knowledge, this is the first report identifying these 3 pathogens in the Korean water deer. Our results suggest that the Korean water deer may serve as a major reservoir for these tick-borne pathogens, leading to spread of tick-borne diseases to domestic animals, livestock, and humans. Further studies are needed to investigate their roles in this respect.
Anaplasma/isolation & purification ; Animals ; Bacterial Infections/epidemiology/microbiology/*veterinary ; Cluster Analysis ; Coinfection/epidemiology/microbiology/veterinary ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Ehrlichia/*isolation & purification ; Korea/epidemiology ; Molecular Sequence Data ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*isolation & purification ; Ruminants/*microbiology ; Sequence Analysis, DNA ; Theileria/*isolation & purification

Species identification using DNA sequences is the basis for DNA taxonomy. In this study, we sequenced the ribosomal large-subunit RNA gene sequences (3,037-3,061 bp) in length of 13 Chinese Theileria stocks that were infective to cattle and sheep. The complete 28S rRNA gene is relatively difficult to amplify and its conserved region is not important for phylogenetic study. Therefore, we selected the D2-D3 region from the complete 28S rRNA sequences for phylogenetic analysis. Our analyses of 28S rRNA gene sequences showed that the 28S rRNA was useful as a phylogenetic marker for analyzing the relationships among Theileria spp. in ruminants. In addition, the D2-D3 region was a short segment that could be used instead of the whole 28S rRNA sequence during the phylogenetic analysis of Theileria, and it may be an ideal DNA barcode.
Animals ; Base Sequence ; China ; DNA, Ribosomal/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 28S/genetics ; Ruminants ; Sequence Alignment ; Sequence Analysis, DNA/veterinary ; Theileria/*classification/genetics/isolation & purification ; Theileriasis/*parasitology

Theileria (T.) buffeli (formerly T. sergenti/T. orientalis) is the major hemo-protozoan distributed in the Far East Asian countries such as Korea, China and Japan. It is responsible for the clinical symptoms of anorexia, ateliosis, anemia, fever and icterus. It also causes abortion and sudden death under severe cases, resulting in economic losses for many livestock farms. The objective of this study was to analyze the genetic diversity of the major surface protein (Msp) gene in T. buffeli in Holstein in Korea, and we characterized the association of the diversification of the Msp gene and its relationship with the pathogenicity of Theileria. For this, complete blood counts and Theileria PCR sequence analysis were performed from 57 Holstein in Jeju Island. A total of 26 PCR positive Holstein (16 anemic and 10 non-anemic) were then randomly selected based on 18s rRNA sequence typing of the Theileria Msp gene. The DNA sequence of the T. buffeli Msp gene in Holstein showed 99.0%, 99.2%, 99.9%, 99.5%, 98.7%, 98.4% and 98.4% homology with T. sergenti, Theileria spp., T. sergenti, Theileria spp., Theileria spp., Theileria spp. and Theileria spp., respectively. The result showed a genetic variation of 57.7% (type I), 3.8% (type II), 15.4% (type III), 7.7% (type IV), 13.5% (type V) and 1.9% (type VI). Type I is the most frequent type in both anemic and non-anemic Holstein while type II was found in only non-anemic Holstein. This 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 Holstein in Jeju Island.
Anemia ; Anorexia ; Antigenic Variation ; Asian Continental Ancestry Group ; Base Sequence ; Blood Cell Count ; China ; Death, Sudden ; Far East ; Fever ; Genetic Variation ; Humans ; Japan ; Jaundice ; Korea ; Livestock ; Polymerase Chain Reaction ; Sequence Analysis ; Theileria

This study was carried out to investigate fifteen cases of acute lethal infection of calves (< or = 4 months of age) by the protozoan parasite Theileria (T.) annulata in the south of Portugal. Calves developed multifocal to coalescent nodular skin lesions, similar to multicentric malignant lymphoma. Infestation with ticks (genus Hyalomma) was intense. Theileria was seen in blood and lymph node smears, and T. annulata infection was confirmed by isolation of schizont-transformed cells and sequencing of hypervariable region 4 of the 18S rRNA gene. At necropsy, hemorrhagic nodules or nodules with a hemorrhagic halo were seen, particularly in the skin, subcutaneous tissue, skeletal and cardiac muscles, pharynx, trachea and intestinal serosa. Histologically, nodules were formed by large, round, lymphoblastoid neoplastic-like cells. Immunohistochemistry (IHC) identified these cells as mostly CD3 positive T lymphocytes and MAC387 positive macrophages. A marker for B lymphocytes (CD79alphacy) labeled very few cells. T. annulata infected cells in these nodules were also identified by IHC through the use of two monoclonal antibodies (1C7 and 1C12) which are diagnostic for the parasite. It was concluded that the pathological changes observed in the different organs and tissues were caused by proliferation of schizont-infected macrophages, which subsequently stimulate a severe uncontrolled proliferation of uninfected T lymphocytes.
Animals ; Base Sequence ; Cattle ; Cattle Diseases/epidemiology/*parasitology/pathology ; Cell Growth Processes/physiology ; DNA, Protozoan/chemistry/genetics ; Disease Outbreaks/*veterinary ; Female ; Immunohistochemistry/veterinary ; Lymphocytes/parasitology/*pathology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; Portugal/epidemiology ; RNA, Ribosomal, 18S/chemistry/genetics ; Sequence Analysis, DNA ; Skin Diseases/epidemiology/parasitology/pathology/*veterinary ; Theileria annulata/*isolation & purification ; Theileriasis/epidemiology/parasitology/*pathology

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.
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

Theileria annulata, a protozoan parasite of cattle and domestic buffaloes, is transmitted by ticks of the genus Hyalomma, and causes a disease named Mediterranean or tropical theileriosis. In this research 50 cattle naturally infected with Theileria annulata were treated with the extract of the plant Peganum harmala. The treatment was continued for 5 days, the dose of the extract being 5 mg/kg per day. After the treatment, 39 cattle responded to the treatment and recovered, but 11 did not respond to the treatment and died. The recovery rate of animals treated with the extract of the plant Peganum harmala was 78%.
Animals ; Antiprotozoal Agents/*therapeutic use ; Cattle ; Lymph Nodes/parasitology/pathology ; *Peganum ; Phytotherapy/*veterinary ; Plant Extracts/*therapeutic use ; *Theileria annulata ; Theileriasis/*drug therapy/pathology ; Treatment Outcome

This study was performed to investigate the prevalence of theileriosis and to compare the prevalence of this disease in Korean native cattle reared under different environmental conditions, namely, in a grazing area and a non-grazing area by polymerase chain reaction. Three hundred and one Korean native cattle (276 cows and 25 bulls) that had not received prior treatment or been vaccinated to prevent theileriosis were examined by PCR for Theileria sergenti infection from 2001 to 2002. In our study, the parasitemia range in T. sergenti-positive cattle by microscopy were from 0.1 to 3% (mean 0.8%). In terms of mean prevalence, 204 of the 301 Korean native cattle (67.8%) were positive reaction by PCR. Our results also revealed that the infection rate among cows (70.3%) was significantly higher than that among bulls (40.0%) (p < 0.01). T. sergenti infection among the over 3 year-old-group (75%) had a significant higher prevalence than that among the less than 3 year-old-group (61.8%) (p < 0.05). Our data also showed that grazing areas (76.1%) had the significant higher prevalence than non-grazing areas (51%) (p < 0.001). In conclusion, this study demonstrates that the prevalence of T. sergenti infection is high and that its prevalence in grazing cattle is higher than that in non-grazing cattle. Therefore, life-long treatment and the development of an optimal vaccine are needed to reduce the numbers of bovine theileriosis in both grazing and non-grazing areas.
Animals ; Cattle ; Cattle Diseases/diagnosis/*epidemiology/parasitology ; DNA, Protozoan/*analysis ; Female ; Korea/epidemiology ; Male ; Poaceae ; Polymerase Chain Reaction/methods/veterinary ; Prevalence ; Sex Factors ; Theileria/genetics/*isolation & purification ; Theileriasis/diagnosis/*epidemiology/parasitology