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Differential responses of monocyte-derived macrophages from Theileria orientalis infected carrier cattle to Pasteruella multocida B:2 infection and latex beads: A preliminary study

Agina OA, Shaari MR, Isa NMM, Ajat MMM, Zamri-Saad M, Samad MJ, et al.

Res Vet Sci, 2023 Dec;165:105073.
PMID: 37939633 DOI: 10.1016/j.rvsc.2023.105073

This study aims to evaluate the responses of peripheral blood monocyte-derived macrophages (PBMDMs) from Theileria orientalis carrier cattle following exposure to Pasteruella multocida B:2 (PM B:2) and latex beads. Twenty-six male crossbred Kedah-Kelantan (KK) cattle were sampled for this study and quantitative PCR (qPCR) was employed in the detection of T. orientalis MPSP gene. Bactericidal assay using a 10:1 multiplicity of infection was performed to measure the phagocytosis and intracellular killing of PM B:2 by PBMDMs. The cell cultures were inoculated with 107 cfu/mL of PM B:2 and incubated in a humidified incubator. The absence of clinical signs, previous history of T. orientalis infection and an MPSP gene copy number below 15,000 GC/μL suggest that the cattle were asymptomatic chronic carriers. A non-significant phagocytic and mean cell death rates were observed in the PBMDMs of T. orientalis positive cattle relative to clinically healthy cattle (CHC) (p > 0.05). The PBMDMs of T. orientalis positive cattle had the lowest mean rate of intracellular killing relative to the CHC at the 30th minute post-infection only (p

Matched MeSH terms: Theileriasis*; Theileria*
Molecular detection, prevalence and risk factors of Theileria orientalis infection among cattle in Peninsular Malaysia

Ola-Fadunsin SD, Sharma RSK, Abdullah DA, Gimba FI, Jesse FFA, Sani RA

Prev Vet Med, 2020 Jul;180:105027.
PMID: 32442824 DOI: 10.1016/j.prevetmed.2020.105027

There is need to confirm the presence of Theileria orientalis among the cattle population in Peninsular Malaysia and to evaluate the risk factors associated with the infection. To this effect, blood samples were collected from 1045 cattle from 43 farms throughout the entire States of Peninsular Malaysia. The collected blood samples were subjected to DNA extraction and subsequent PCR amplification of the major piroplasm surface protein (MPSP) gene of the haemoprotozoan. Representative positive amplicons were purified, sequenced and compared with other sequences of the MPSP gene of T. orientalis curated from the GenBank. A well-structured questionnaire was used to get information about each cattle, it's demography, the bio-security, environmental and management factors. Univariate and multivariate analysis were used for the statistical evaluation, with significance set at p < 0.05. A total prevalence of 49.76% (520/1045; 95% CI: 46.73 - 52.79) was obtained. Types of breeds, age, production type, herd size, level of farm biosecurity, farm size, presence of other animal species in the farm, management systems and prophylaxis were significantly (p < 0.05) associated with the prevalence of T. orientalis. This study confirmed the presence of T. orientalis and establish that the haemoprotozoan is endemic among cattle in Peninsular Malaysia.

Matched MeSH terms: Theileriasis/epidemiology*; Theileriasis/parasitology; Theileria/isolation & purification*
Molecular phylogenetic studies on Theileria parasites based on small subunit ribosomal RNA gene sequences

Chansiri K, Kawazu S, Kamio T, Terada Y, Fujisaki K, Philippe H, et al.

Vet Parasitol, 1999 Jun 15;83(2):99-105.
PMID: 10392966

Classification of Theileria parasites of south-east Asian countries is still ambiguous due to the lack of basic studies, especially their molecular genetic information. In this study, we included 6 known species and 14 unclassified Theileria parasite isolates: Theileria annulata, Theileria parva, Theileria taurotragi, Theileria sergenti, Theileria buffeli, Theileria types Sable, Theileria types A, B, B1, B2, C, D, E, F, G, G1, Theileria type Medan (Indonesia), Theileria type Ipoh (Malaysia) and Theileria type Thong Song (Thailand). Small subunit ribosomal RNA (srRNA) nucleotide sequence data were collected by PCR, cloning and dideoxy sequencing. The srRNA nucleotide sequences were aligned and analyzed by distance methods, maximum parsimony algorithms and maximum likelihood methods to construct phylogenetic trees. Bootstrap analysis was used to test the strength of the different phylogenetic reconstructions. The data indicated that all of the tree-building methods gave very similar results. This study identified two groups of Theileria, the pathogenic and benign groups, which are strongly supported by bootstrap analysis. The analysis also indicated that three subgroups (A, B and C) were generated within the benign Theileria group whereas the classification of Theileria type D and Thong Song is questionable. However, more basic information such as life cycle differences, vectors, modes of transmission, virulent and genetic/sexual compatability is essential for clearer taxonomic definition of the benign Theileria parasites.

Matched MeSH terms: Theileriasis/parasitology; Theileria/classification; Theileria/genetics*
Genetic characterization of Theileria species infecting bovines in India

Kundave VR, Ram H, Shahzad M, Garg R, Banerjee PS, Nehra AK, et al.

Infect Genet Evol, 2019 11;75:103962.
PMID: 31302242 DOI: 10.1016/j.meegid.2019.103962

Genetic characterization of Theileria species infecting bovines in India was attempted targeting the 18S ribosomal RNA region of the parasite. Blood samples of bovines (n = 452), suspected for haemoprotozoan infections, from 9 different states of the country were microscopically examined for Theileria species infection. Four Theileria spp. positive blood samples from each state were randomly utilized for PCR amplification of the 18S rRNA gene (approx. 1529 bp) followed by cloning and sequencing. The sequence data analysis of all the 36 isolates revealed that 33 isolates had high sequence similarity with published sequences of T. annulata, whereas 3 isolates (MF287917, MF287924 and MF287928) showed close similarity with published sequences of T. orientalis. Sequence homology within the isolates ranged between 95.8 and 100% and variation in the length of targeted region was also noticed in different isolates (1527-1538 nt). Phylogenetic tree created for T. annulata sequences revealed that a total of 24 Indian isolates formed a major clade and grouped together with isolates originating from countries like China, Spain, Turkey and USA. Remaining 09 isolates clustered in a separate group and were closely related to the TA5 isolate of T. annulata (a new genotype) originating from India and also with the isolates from East Asian countries like Japan and Malaysia. All the three T. orientalis isolates had minimal intraspecific variation (99-100% homology) amongst themselves. Further, in the phylogenetic analysis T. orientalis Indian isolates were found to cluster away from other 14 isolates of T. buffeli/sergenti/orientalis originating from different countries (Australia, China, Indonesia and Spain). However, these 3 isolates clustered together with the T. buffeli Indian isolate (EF126184). Present study confirmed the circulation of different genotypes of T. annulata in India, along with T. orientalis isolates.

Matched MeSH terms: Theileriasis/epidemiology; Theileriasis/parasitology*; Theileria/genetics*
Fulltext Molecular detection of Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos, Trypanosoma evansi and first evidence of Theileria sinensis-associated bovine anaemia in crossbred Kedah-Kelantan x Brahman cattle

Agina OA, Shaari MR, Isa NMM, Ajat M, Zamri-Saad M, Mazlan M, et al.

BMC Vet Res, 2021 Jul 18;17(1):246.
PMID: 34275459 DOI: 10.1186/s12917-021-02902-0

BACKGROUND: Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos and Trypanosoma evansi, and determine their phylogenetic relationships and haemato-biochemical abnormalities in naturally infected cattle.
METHODS: Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of Anaplasma and Mycoplasma species, MPSP genes of T. orientalis and T. sinensis, MSP4 gene of A. marginale, 16SrRNA gene of Candidatus Mycoplasma haemobos, and RoTat1.2 VSG gene of Trypanosoma evansi, in sixty-one (61) clinically ill Kedah-Kelantan x Brahman cattle in Pahang, Malaysia.
RESULTS: A total of 44 (72.13%) cattle were infected with more than one blood pathogen. Theileria species was the blood pathogen with the highest molecular detection rate (72.13, 95% CI 59.83-81.81%). Nucleotide blast analyses of all sequences demonstrated high degree of molecular similarity (98-100%) in comparison with their respective reference sequences. Analysis of 18SrRNA gene sequences of Theileria species and 16SrRNA gene sequences of Anaplasma species revealed Theileria sinensis and Anaplasma platys respectively as additional species detected in these cattle. MPSP-PCR analysis was conducted for further confirmation of T. sinensis. The blood picture of eight infected cattle groups revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. High mean erythrocyte fragility values were common in infected cattle groups. Anaemia of the macrocytic normochromic type and spherocytes were observed in the T. evansi and Anaplasma platys + Theileria sinensis double species co-infected cattle group. Normocytic normochromic anaemia was observed in the T. sinensis infected cattle group. Significant (p Theileria sinensis-associated bovine anaemia (TSABA) in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of A. platys, there is an urgent need for appropriate preventive and control measures against these blood pathogens.

Matched MeSH terms: Theileriasis/blood; Theileriasis/epidemiology*; Theileria/genetics; Theileria/isolation & purification
Fulltext The prevalence of parasitic infestation of small ruminant farms in perak, malaysia

Zainalabidin FA, Raimy N, Yaacob MH, Musbah A, Bathmanaban P, Ismail EA, et al.

Trop Life Sci Res, 2015 Apr;26(1):1-8.
PMID: 26019746 MyJurnal

Helminthiasis due to strongyles such as Haemonchus contortus, coccidiosis caused by Eimeria sp. and blood parasite diseases such as theileriosis by Theileria sp. have been reported to cause severe morbidity and mortality annually in small ruminants in Malaysia. The aims of this study were to investigate the prevalence of helminthiasis, coccidiosis and theileriosis and to determine the packed cell volume (PCV) value of small ruminants in Perak, Malaysia. Blood and faecal samples were obtained from a total of 175 animals from 7 small ruminant farms in Kampar, Larut Matang and Selama, Kuala Kangsar and Manjung districts in Perak; the samples were examined for parasitic infestations from April to July 2011. The results of this study show that H. contortus was found in 152 (86.86%) animals, Eimeria sp. was found in 162 (92.57%) animals and the blood protozoa Theileria sp. was found in 25 (14.30%) animals. The PCV values of all of these animals were recorded between 7% and 44%. A total of 42 (24%) animals were anaemic, with a PCV of less than 21%. Continuous monitoring of small ruminant farms will provide important information for assisting farmers with managing the spread of parasitic infections and maintaining the productivity of animals.

Matched MeSH terms: Theileriasis; Theileria
The first molecular survey of theileriosis in Malaysian cattle, sheep and goats

Kho KL, Amarajothi ADG, Koh FX, Panchadcharam C, Hassan Nizam QN, Tay ST

Vet Parasitol Reg Stud Reports, 2017 12;10:149-153.
PMID: 31014589 DOI: 10.1016/j.vprsr.2017.08.003

This study reports the molecular detection of Theileria spp. from six cattle farms, a sheep farm and a goat farm located at different states in Peninsular Malaysia. Animal blood samples were screened for the presence of Theileria DNA using a conventional polymerase chain reaction (PCR) assay. A total of 155 (69.2%) of 224 cattle investigated were PCR-positive for Theileria DNA. The occurrences of Theileria spp. ranged from 17.5% to 100.0% across six cattle farms. Theileria DNA was detected from 90.0% of 40 sheep but none of 40 goats examined in this study. Sequence analyses of amplified 18S rRNA partial fragments (335-338bp) confirmed the identification of Theileria buffeli, Theileria sergenti, and Theileria sinensis in representative samples of cattle and ticks. T. luwenshuni was identified in the infected sheep. The high occurrences of Theileria spp. in our farm animals highlight the needs for appropriate control and preventive measures for theileriosis.

Matched MeSH terms: Theileriasis/blood; Theileriasis/epidemiology*; Theileria/isolation & purification