Genetic Diversity of Benign Theileria Parasites of Cattle in the Okinawa Prefecture

Zakimi, Satoshi; Kim, Jung-Yeon; Oshiro, Mamoru; Hayashida, Kentaro; Fujisaki, Kozo; Sugimoto, Chihiro

doi:10.1292/jvms.68.1335

Cited by 33 publications

(41 citation statements)

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“…The cow (an Angus) from the farm in Shintoku district that harbored T. orientalis with the type-5 MPSP gene was originally born on the farm, but its ancestors had been introduced from Miyazaki and Kagoshima prefectures, located in Kyushu Island, in the southern part of Japan. Type-4 and -5 MPSP genes have previously been found in Okinawa prefecture [27], and were also detected in the Kyushu area in our latest study (manuscript in preparation). In Shin-Hidaka district, the cow (a Hereford) with the type-4 MPSP and Buffeli-type p23 genes was also born on the study farm.…”

Section: Discussionsupporting

confidence: 71%

“…Blood plasma fractions were obtained by centrifugation at 700  g for 15 min at 4C, and then stored at -30C, for subsequent analysis by enzyme-linked immunosorbent assay (ELISA). PCR: A pair of universal primers (857 base pairs (bp)) [8,24], and three kinds of type-specific primer pairs for the Ikeda strain (826 bp), Chitose strain (831 bp), and T. buffeli Warwick stock (825 bp) [10,13,27] were used for the PCR identification of T. orientalis. All the primer pairs were targeted to the MPSP (p32/33/34) gene of T. orientalis, and were able to amplify the indicated sizes of DNA fragments by PCR.…”

Section: Collection Of Blood Samplesmentioning

confidence: 99%

“…The MPSP and p23 genes are expressed in the intraerythrocytic stage of the parasite, called piroplasm, and are conserved to some degree among field isolates of T. orientalis [6,8,17]. T. orientalis populations have recently been found to consist of seven types, based on a series of all the registered MPSP gene sequences [10,27]. Another gene, which encodes a 23-kDa piroplasm membrane protein, was also reported to show genetic diversity [18,28].…”

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Epidemiological Survey of Theileria orientalis Infection in Grazing Cattle in the Eastern Part of Hokkaido, Japan

et al. 2009

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ABSTRACT. Theileria orientalis is one of the benign species of Theileria that is widely distributed in Japan and is sometimes responsible for serious economic losses in the livestock industry. In the present study, we surveyed the current status of T. orientalis infection in grazing cattle in the eastern areas of Hokkaido (Taiki, Otofuke, Shintoku, and Shin-Hidaka districts) using molecular methods, as well as traditional methods, of diagnosis. The genes encoding the major piroplasm surface protein (MPSP) and p23 of T. orientalis were identified using highly detectable polymerase chain reaction (PCR). Results of the MPSP-PCR assay indicated that grazing cattle in these districts, after about 1.5 months pasturage, showed high rates of infection, ranging from 10.0-64.8%. Although the main MPSP and p23 genotypes detected were the Ikeda-or Chitose-types, an MPSP gene closely relating to that found in Okinawa prefecture, and a p23 gene closely relating to the Australian (Warwick) Buffeli-type gene, were found in the cattle in Shintoku and Shin-Hidaka districts. The present survey indicated that there were at least five types of T. orientalis classified by their MPSP genes in Hokkaido, Japan, and that T. orientalis infection rates are still high in this region.

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confidence: 71%

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Epidemiological Survey of Theileria orientalis Infection in Grazing Cattle in the Eastern Part of Hokkaido, Japan

et al. 2009

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“…Of these genotypes, type 2 (Ikeda) and to a lesser extent type 1 (Chitose) are typically found in association with clinical disease (6,9,10,(17)(18)(19)(20)(21)(22). The presence of pathogenic and benign forms of T. orientalis greatly complicates its clinical diagnosis, with standard blood film analysis unable to identify the pathogenic genotypes.Multiple conventional PCR (cPCR) assays have been published for the identification of T. orientalis in blood samples (9,21,23,24). The most commonly cited assays detect and genotype T. orientalis by amplifying unique regions of the MPSP gene (21).…”

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confidence: 99%

“…These assays use two universal primers to detect T. orientalis infection and specific forward primers to identify the Ikeda, Chitose, and Buffeli genotypes. While this method is highly sensitive and has been validated in prior studies (19,21), it is not multiplexed and is highly susceptible to PCR inhibitors (6), which are often found in nucleotide extractions obtained from blood (25,26). To overcome inhibition, undiluted and diluted nucleotide extracts can be examined in parallel to prevent false-negative results (6,19,27).…”

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Development and Validation of a Quantitative PCR Assay Using Multiplexed Hydrolysis Probes for Detection and Quantification of Theileria orientalis Isolates and Differentiation of Clinically Relevant Subtypes

et al. 2015

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T heileria orientalis is a vector-borne hemoprotozoan that infects cattle and buffalo and is generally spread by ticks of the Haemaphysalis genus (1-3). Historically, this organism has been referred to as Theileria sergenti, Theileria buffeli, or the T. orientalis/T. sergenti/T. buffeli complex; however, the name T. sergenti is now considered invalid (4), and T. orientalis is commonly used to refer to all (5). From a clinical perspective, T. orientalis can cause anemia, lethargy, jaundice, fever, abortion, and mortality in cattle (6). Clinical infection can also result in decreased milk production in dairy cattle (7). T. orientalis is a conditional pathogen, and while pathogenic forms are largely limited to Eastern Asia and Australasia (5, 6, 8-10), it is frequently detected in asymptomatic animals; these benign forms are globally spread (5,(11)(12)(13)(14). Analysis of the most common genotyping locus (p32), encoding the variable major piroplasm surface protein (MPSP), currently identifies 11 distinct T. orientalis genotypes (13,15,16). Of these genotypes, type 2 (Ikeda) and to a lesser extent type 1 (Chitose) are typically found in association with clinical disease (6,9,10,(17)(18)(19)(20)(21)(22). The presence of pathogenic and benign forms of T. orientalis greatly complicates its clinical diagnosis, with standard blood film analysis unable to identify the pathogenic genotypes.Multiple conventional PCR (cPCR) assays have been published for the identification of T. orientalis in blood samples (9,21,23,24). The most commonly cited assays detect and genotype T. orientalis by amplifying unique regions of the MPSP gene (21). These assays use two universal primers to detect T. orientalis infection and specific forward primers to identify the Ikeda, Chitose, and Buffeli genotypes. While this method is highly sensitive and has been validated in prior studies (19,21), it is not multiplexed and is highly susceptible to PCR inhibitors (6), which are often found in nucleotide extractions obtained from blood (25,26). To overcome inhibition, undiluted and diluted nucleotide extracts can be examined in parallel to prevent false-negative results (6,19,27). However, if multiple reactions per sample are required to determine the presence of infection, the procedure becomes both expensive and time-consuming. Furthermore, cPCR does not give an accurate representation of parasite load and therefore cannot provide an indication of the severity of T. orientalis infection. A recently developed multiplexed tandem PCR (28) is able to discriminate between four genotypes of T. orientalis, but it is only semiquantitative and therefore cannot be used to define a clinical threshold.Hydrolysis probe quantitative PCR (qPCR) assays employ sequence-specific, fluorescently labeled probes attached to duplex-

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Mutation scanning‐based analysis of Theileria orientalis populations in cattle following an outbreak

Cufos¹,

Jabbar

Carvalho

et al. 2012

Electrophoresis

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Bovine theileriosis is a tick-borne disease caused by one or more hemoprotozoan parasites of the genus Theileria. In the past, Theileria infection in cattle in Australia was largely asymptomatic and recognized to be associated with Theileria buffeli. However, outbreaks of theileriosis have occurred in beef and dairy cattle in subtropical climatic regions (New South Wales) of Australia. There is also one published report of a recent theileriosis outbreak in a beef farm near Seymour in the southeastern state of Victoria. In order to gain an improved insight into the genetic composition of Theileria populations following this outbreak, we undertook herein an integrated PCR-coupled mutation scanning-sequencing-phylogenetic analysis of sequence variation in part of the major piroplasm surface protein (MPSP) gene within and among samples from cattle involved in the outbreak. Theileria DNA was detected in 89.4% of 94 cattle in the Seymour farm; the genetic analysis showed that the ikeda and chitose genotypes representing the Theileria orientalis complex were detected in 75 and 4.8% of 84 infected cattle, respectively, and that mixed populations of these two genotypes were found in 20.2% of infected cattle. Given unpublished reports of a significant increase in the number of outbreaks in Victoria, future investigations should focus sharply on elucidating the epidemiology of Theileria to subvert the economic impact on the cattle industry in this state. Although used here to explore genetic variation within the T. orientalis complex in Australia, a mutation scanning-based approach has broad applicability to other species of Theileria in other countries.

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Genetic Diversity of Benign Theileria Parasites of Cattle in the Okinawa Prefecture

Cited by 33 publications

References 18 publications

Epidemiological Survey of Theileria orientalis Infection in Grazing Cattle in the Eastern Part of Hokkaido, Japan

Epidemiological Survey of Theileria orientalis Infection in Grazing Cattle in the Eastern Part of Hokkaido, Japan

Development and Validation of a Quantitative PCR Assay Using Multiplexed Hydrolysis Probes for Detection and Quantification of Theileria orientalis Isolates and Differentiation of Clinically Relevant Subtypes

Mutation scanning‐based analysis of Theileria orientalis populations in cattle following an outbreak

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