Genetic diversity of Theileria orientalis from cattle in Turkey

Theileria orientalis is usually a benign parasite but some genotypes cause infection and economic losses to the cattle industry. This study was carried out to determine T. orientalis genotypes in cattle. T. orientalis positive 63 sample were analyzed by amplifying the MPSP gene region by PCR. As a result of the SSCP analysis, samples with different band profiles were sent to the sequence analysis and genotypes were determined. T. orientalis genotype-specific PCR was performed to determine the mix genotypes. Type 1 (chitose), type 3 and type 1-type 3 mix were found positive 11.1%, 46%, and 17.5% respectively. In addition, phylogenetic analysis was performed to separate the chitose genotypes, and two samples were found in chitose A, one sample was found in chitose B. Although chitose A genotype is suggested to be more pathogenic than chitose B, but there is little evidence for this. As a result of this study, we showed the presence of pathogenic genotype T. orientalis in Turkey. Therefore, extensive epidemiological studies are required to understand the geographic distribution, different genotypes and clinical pathologies of T. orientalis.     

Genetic diversity of major piroplasm surface protein genes and their allelic variants of Theileria parasites in Thai cattle.

Twenty-eight field isolated Theileria parasite DNAs obtained from dairy and beef cattle in distinct geographical areas of Thailand were characterized by using polymerase chain reaction (PCR) amplification with six sets of oligonucleotide primers. Three sets of them were modified from two genes of immunodominant major piroplasm surface protein (MPSP) coding for 32 kDa (p32) of T. sergenti and 33/34 kDa (p33/34) of T. buffeli, and MPSP of Theileria spp.(Thai-isolate). The other three sets of primers were basically generated from three alleles of MPSP which were specific for Japanese T. sergenti-Ikeda stock (I-type), Japanese T. sergenti-Chitose stock (C-type) and Australian T. buffeli-Warwick stock (B1-type), respectively. The results indicated that 14 out of 28 isolates were amplified by the Thai-specific primer whereas 6 isolates were amplified by the p32 specific primer and the other 5 isolates were amplified by the p32 and Thai-specific primers. In addition, by using the allele-specific PCR, 14 out of 28 isolates contained C-type MPSP whereas 3 isolates contained B1 type parasites. Interestingly, 20 out of 28 isolates could be amplified by the Thai-specific primer. The majority of Theileria parasites distributed in Thailand contained Thai type parasites, whereas C-type parasites showed the mixed population with B1 and Thai type parasites. No I type parasite was detected.     

Genotypic diversity of Theileria orientalis detected from cattle grazing in Kumamoto and Okinawa prefectures of Japan

Theileria orientalis is a benign protozoan species that is widely distributed in Japan, yet sometimes causes serious economic losses in the livestock industry. In this study, we conducted a molecular survey based on genes encoding the major piroplasm surface protein (MPSP) and p23 for T. orientalis detected in cattle grazing in southern areas of Japan, consisting of 2 farms in Kumamoto prefecture (Aso and Kuma districts) and 3 farms in Okinawa prefecture (Ishigaki, Iriomote, and Yonaguni Islands). High prevalence rates of T. orientalis infection were shown in all the cattle populations using the diagnostic MPSP- and p23-PCR assays. Phylogenetic analyses revealed 4 MPSP genotypes and 3 p23 genotypes. Furthermore, MPSP genotype-specific PCR methods were developed in this study and wide distributions of 5-district genotypes of T. orientalis were observed for the examined farms. Our results indicate that at least 5 types of T. orientalis exist in Kumamoto and Okinawa prefectures of Japan and that genotype-specific PCR assays are highly applicable for the quarantine of transported cattle and for epidemiological surveys of bovine theileriosis in Japan.     

Genetic diversity and genomic inbreeding in Japanese Black cows in the islands of Okinawa Prefecture evaluated using single-nucleotide polymorphism array

Maintaining genetic diversity and inbreeding control are important in Japanese Black cattle production, especially in remote areas such as the islands of Okinawa Prefecture. Using a single-nucleotide polymorphism (SNP) array, we evaluated the genetic diversity and genomic inbreeding in Japanese Black cows from the islands of Okinawa Prefecture and compared them to those from other locations across Japan. Linkage disequilibrium decay was slower in cows in the islands of Okinawa Prefecture. The estimated effective population size declined over time in both populations. The genomic inbreeding coefficient (F_ROH) was estimated using long stretches of consecutive homozygous SNPs (runs of homozygosity; ROH). F_ROH was higher in the cows on the islands of Okinawa Prefecture than on other locations. In total, 818 ROH fragments, including those containing NCAPG and PLAG1, which are major quantitative trait loci for carcass weight in Japanese Black cattle, were present at significantly higher frequencies in cows in the islands of Okinawa Prefecture. This suggests that the ROH fragments are under strong selection and that cows in the islands of Okinawa Prefecture have low genetic diversity and high genomic inbreeding relative to those at other locations. SNP arrays are useful tools for evaluating genetic diversity and genomic inbreeding in cattle.     

Survey of benign Theileria parasites of cattle and buffaloes in Thailand using allele-specific polymerase chain reaction of major piroplasm surface protein gene

During a year from 1999 to 2000, a total of 247 blood samples were collected from 214 cattle and 33 water buffaloes in 16 distinct geographical locations of Thailand and analyzed by allele-specific PCR amplification of major piroplasm surface protein (MPSP) genes of benign Theileria parasites. Four allelic MPSP gene types were determined namely C-type, I-type, B-type and Thai-type, which were originally designated from Japanese Theileria orientalis (Chitose, Ikeda), Australian T. buffeli (Warwick) and Thai T. sp. (Kamphaeng Saen), respectively. Only two allelic MPSP gene types were successively amplified from 204 (82.6%) blood samples. Among positive cases, 138 (67.6%) and 17 (8.3%) samples contained either Thai-type or C-type parasites, respectively, while 49 (24%) samples contained both types. However, nucleotide sequences of MPSP genes of Thai T. sp. amplified by C-type specific primers revealed higher (96.3%) similarity to Indonesian T. sp. rather than (87.8% similarity) to Japanese T. orientalis (Chitose) designated as C-type.     

Genetic diversity in Swiss cattle breeds

Introduction First attempts at establishing the genetic relationships among cattle populations relied on archeological evidence (Epstein 1971; Epstein and Mason 1984) and protein polymorphisms (Baker and Manwell 1980, 1991). Loftus et al. (1994) examined mitochondrial DNA to determine the divergence time between Bos taurus and Bos indicus. Today most studies on genetic diversity are based on microsatellite analysis (Litt and Luty 1989; Tautz 1989; Weber and May 1989). Microsatellites were used in, e.g. man (Bowcock et al. 1994), canids (Roy et al. 1994; Fredholm and Winter ø ; 1995) and sheep (Buchanan et al. 1994). Recent studies in cattle are also microsatellite based (e.g. Machugh et al. 1994; Ciampolini et al. 1995; Moazami -Goudarzi et al. 1997) and aim at facilitating the development of management programs for endangered breeds (FAO 1981). Our microsatellite-based investigation on the genetic diversity between and within Swiss cattle breeds included Original Swiss Brown, purebred Simmental, Holstein, Hérens and Evolènard. Previous studies in Swiss breeds made use of blood group systems (Reuse 1969), serum transferrin and hemoglobin (Krummen 1964), amylase (Buser 1970) and carboanhydrases (Kä ;stli et al. 1980). The Hérens breed is endemic to the canton of Wallis. The Evolènard, which are very few in numbers and restricted to a single valley in the canton of Wallis, are phenotypically very similar to the Hérens with the exception of the coat colour. In the Aosta valley (Italy) which borders the canton of Wallis these two breeds find their counterparts. The phenotype of the Aosta Chestnut fits the Hérens and the Aosta Black Pied fits the Evolènard. The Holstein breed replaced the Fribourg breed which was a colour variant of the purebred Simmental breed (Engeler et al. 1961) and is now extinct. The Original Swiss Brown and the purebred Simmental are endemic to Switzerland.     

Molecular epidemiological survey of benign Theileria parasites of cattle in Japan: detection of a new type of major piroplasm surface protein gene

Benign Theileria species of cattle are found in most parts of the world. The major piroplasm surface protein (MPSP), a conserved protein in all Theileria species, has been used as a maker for epidemiological and phylogenetical studies of benign Theileria species. Parasites with Ikeda- or Chitose-type MPSP genes are dominant in Japan, but we report here mixed infection cases of Theileria parasites with an additional MPSP type parasite infecting cattle in Abashiri District, Hokkaido. The MPSP gene sequence found in the additional type was closely related to MPSP genes of Theileria parasites found in Southeast Asian countries, including Thailand (Narathiwat) and Indonesia (Java). Theileria parasites from the blood sample were also distinguishable from the Ikeda or Chitose type parasites by the small subunit (SSU) rRNA gene sequence analysis, and they are grouped into the SSU rRNA types C/D found in Korea, North America, and Spain. The present finding of mixed infections of cattle with three different types of Theileria makes epidemiological feature of bovine theileriosis in Japan more complex. We have designed a set of primers specific to this MPSP type in order to conduct further epidemiological study.     

Antigenic diversity of Theileria major piroplasm surface protein gene in Jeju black cattle

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.     

Genetic diversity of Afrikaner cattle in southern Africa

The Afrikaner is an indigenous South African breed of “Sanga” type beef cattle along with breeds such as the Drakensberger and Nguni. Six composite breeds have been developed from crosses with the Afrikaner. Additionally, Afrikaner has been the base from which exotic breeds were established in South Africa through backcrossing. The study examined genetic diversity of Afrikaner cattle by genotyping 1257 animals from 27 herds in different geographic areas of South Africa and Namibia using 11 microsatellite markers. Multiple-locus assignment, performed using the Bayesian clustering algorithm of STRUCTURE, revealed three underlying genotypic groups. These groups were not geographically localized. Across herds and markers, the proportion of unbiased heterozygosity ranged from 0.49 to 0.72 averaging 0.57; mean number of alleles per locus ranged from 3.18 to 7.09, averaging 4.81; and allelic richness ranged from 2.35 to 3.38, averaging 2.67. It is concluded that a low inbreeding level of 2.7% and a moderate to high degree of variation still persists within the Afrikaner cattle breed, despite the recent decline in numbers of animals.     

Genetic diversity among some cattle breeds in the Alpine area

Seven autochthonous Italian cattle breeds bred in the alpine area (Aosta Black Pied, Aosta Red Pied, Aosta Chestnut, Oropa Red Pied, Grey Alpine, Rendena and Burlina) were investigated in order to characterise their genetic structure and to study their phylogenetic origin. Two cattle breeds from Germany (Original German Brown and Holstein) and four from Switzerland (Simmental, Herens, Evolene and Brown Swiss) were included in the study in order to determine the genetic diversity existing among Italian local breeds, similar breeds bred on the other side of the Alps and in the well known Holstein.
Seventeen microsatellites, of the internationally accepted panel for the study of cattle biodiversity, were used for the analysis.
Microsatellites were highly polymorphic with a mean number of 5,5 alleles (ranging from 2 to 12 per locus). For each locus, allelic frequencies, heterozygosity (H) and the Polymorphism Information Content (PIC) were computed. The genetic equilibrium according to Hardy–Weinberg was calculated for each population and for each locus. Allele frequencies were used to estimate genetic distances and to draw a phylogenetic tree. The two closest breeds were Aosta Red Pied and Aosta Black Pied, while the two genetically most different were Holstein and Aosta Chestnut. Aosta valley breeds, Evolene and Herens constituted a tight cluster in the phylogenetic consensus tree. Principal component analysis showed a similar pattern for all the alpine breeds, while Holstein and Original German Brown were far away. The genetic differences among breeds were in accordance with their geographical and historical origins.     

Genetic diversity of growth hormone receptor gene in cattle

Growth hormone receptor (GHR) belongs to a member of the cytokine receptor superfamily. Polymorphism of presence or absence of an approximately 1.2 kbp LINE-1 element is observed in bovine GHR gene. The present study was carried out for estimating the genetic diversity and the origin of the LINE-1 element in 10 European, Southeastern Asian and East Asian cattle breeds or populations. Genotyping of the LINE-1 revealed predominant LINE-1 presence in European breeds (0.917∼0.991), absence in the Bos taurus indicus populations (0.000∼0.017), and intermediate presence in Northeast Asian cattle (0.417∼0.522). From genetic features of LINE families, LINE-1 of GHR could be attributed to the same origin in both European and Asian cattle, and Asian LINE-1 may not be derived from recent introgression. This result suggested that LINE-1 in bovine GHR gene could have arisen in an ancestral population of Bos taurus taurus .     

甘孜藏牛mtDNA 基因组遗传多样性分析

［目的］探究甘孜藏牛的mtDNA基因组遗传多样性与母系起源。［方法］采用mtDNA全基因组序列比对及生物信息学方法。［结果］结果显示：在28头甘孜藏牛mtDNA 基因组中,共检测到1232个变异位点,确定了22种单倍型,其单倍型多样度（Hd）为0.98820±0.00010,核苷酸多样度（Pi）为0.02420±0.00003,表明甘孜藏牛具有丰富的母系遗传多样性。系统发育树和网络分布图表明,28头甘孜藏牛mtDNA基因组包括4种母系支系,分别为普通牛的T2、T3与T4支系,还有牦牛支系,其中T2支系占7.14%,T3支系占64.29%,T4支系占3.57%,牦牛支系占25 %。［结论］甘孜藏牛具有较丰富的母系遗传多样性,为普通牛母系起源,但与牦牛有杂交。     

Theileria mutans infection in nigerian cattle

Summary In a 12 year survey in the former Northern Provinces of Nigeria antibodies toTheileria mutans were found to have a prevalence rate of 32.6 to 85%. The indirect fluorescent antibody test was found to be more efficient than thin blood smear examination in detecting cattle exposed toT. mutans. The high prevalence ofT. mutans infection was attributed to the high prevalence ofAmblyomma species in the area under study.

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Infeccion ConTheileria Mutans En Ganado Nigeriano

Resumen En un reconocimiento que duró 12 años, en las otrora Provincias Septrionales de Nigeria, se encontraron anticuerpos deTheileria mutans, con una rata de prevalencia de 32.6 a 85%. La prueba indirecta de anticuerpos fluorescentes, fué más eficiente que los frotis sanguíneos, para detectar animales infectados. La alta prevalencia de infecciones porT. mutans, se atribuyó a la alta prevalencia de especies deAmblyoma en el área de estudio.

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Theileriose AT. Mutans Chez Des Bovins Du Nigeria

Resumé Au cours d'une enquête dans les provinces nord du Nigéria, des anticorps deTheileria mutans ont été trouvés atteignant un taux d'incidence de 32,6 à 85 p.100. L'immunofluorescence indirecte s'est révélée plus efficace que l'examen de frottis de sang sur lame pour mettre en évidence les bovins exposés àTh. mutans. L'incidence élevée de la theilériose àTh. mutans a été atribuée à la densité importante de l'espèceAmblyomma dans la région étudiée.

     

Intraerythrocytic schizogony of Theileria sergenti in cattle

The characteristics of developing intraerythrocytic stages of T. sergenti were studied by light and transmission electron microscopy. The parasites with many ribosomes, acristate mitochondria, cytostome, and food vacuoles were morphologically regarded as the trophozoite stage. Although this type of parasites was frequently detected, intraerythrocytic merozoite stage with electron dense cisternae, rhoptries and small electron dense bodies was rarely observed in high parasitaemia. The intraerythrocytic stages of T. sergenti were divided mainly into four daughters by schizogony, and alternatively into two by binary fission. The daughter parasites in each division had the same ultrastructural features as of merozoites. As a result, it was suggested that T. sergenti trophozoites multiplied by schizogony to four organisms or by binary fission in the peripheral erythrocyte, and differentiated to the merozoites which acquired penetrating ability into the erythrocytes.     

群牛暴发牛泰勒虫病的诊治

牛泰勒虫病在南方地区较为罕见。2001年7月，湖南省衡阳县樟树乡某村牛场所养的93头牛暴发牛泰勒虫病，经笔者诊治及时控制了疫情，现报告如下：     

Isolation and characterization of bovine Theileria parasites in Zimbabwe

Theilerial parasites of cattle were isolated by a variety of methods from the Harare area of Zimbabwe. Parasite stocks were established in lymphoid cell cultures and as cryopreserved sporozoite stabilates in the laboratory. Fourteen stocks in culture were characterized by testing them with monoclonal antibodies (MAb) raised against T. parva parva and T. parva lawrencei antigen. Two of these stocks had profiles similar to T. taurotragi isolates from East Africa, the other stocks had profiles similar to T. parva parva, however, many of them failed to bind MAb No. 7, and this may be a distinctive feature for T. parva bovis. Three T. p. bovis stocks were titrated by injecting different doses of the respective stabilates into pairs of cattle. Reactions ranged from severe to inapparent according to the stocks and dose used, but no fatal reactions were recorded, even at the highest dose rate. On recovery, all cattle were given homologous and then heterologous challenge. The results of the latter challenge showed that the Boleni stock gave good cross-protection against challenge with two other Zimbabwean stocks. This stock may therefore be a candidate for immunizing cattle, under field conditions, to protect them against T. p. bovis in Zimbabwe. Non-pathogenic strains of T. p. bovis may be difficult to distinguish from T. taurotragi unless cross-challenge experiments can be conducted and/or MAb profiles have been made. An improved serological test is needed to differentiate antibodies to these parasites in the sera of recovered cattle.     

Naturally occurring infections of cattle with Theileria lestoquardi and sheep with Theileria annulata in the Sudan

Theileria annulata is endemic in northern Sudan, hindering all efforts at upgrading cattle for milk production. T. lestoquardi clinical cases occur throughout the year and causes annual outbreaks that result in substantial losses in sheep. In the northern Sudan both cattle and small ruminants are frequently raised together and/or share common grazing grounds at river banks. In an attempt to evaluate field cross infectivity of Theileria lestoquardi and T. annulata in cattle and sheep respectively, a PCR analysis was carried out on samples collected from closely reared sheep and cattle using both T. annulata and T. lestoquardi specific primers. A total of 19 sheep out of 51 (37.3%) were positive for T. lestoquardi while four sheep (7.8%) showed T. annulata specific amplicons. A total of 38 out of 52 (73.1%) surveyed cattle were PCR positive for T. annulata and only two (3.8%) showed T. lestoquardi specific bands. These findings indicate complex epidemiology of both infections in areas where both parasites are transmitted by the same vector and call for further investigations of this phenomenon.     

Antigenic diversity of Theileria annulata macroschizonts

A series of monoclonal antibodies has been produced which reacts with the intracellular macroschizont of Theileria annulata, and this series has been used to examine the level of antigenic diversity between and within stocks of the parasite in addition to species specificity within the genus. The majority of the monoclonal antibodies (six of eight) reacted with all stocks tested and four were species-specific. Two monoclonal antibodies detected variation between stocks and within stocks. The variation within a stock was manifest as only a proportion of the macroschizonts reacting with the monoclonal antibody. This was demonstrated as being due to the strain containing more than one antigenic type of parasite by cloning the infected lymphocyte cell line and showing that the resulting cloned cell lines were antigenically homogeneous. The variation between stocks allowed them to be divided into three groups on the basis of antigenic profile.