BACKGROUND: Ovine theileriosis is an important hemoprotozoal disease of sheep and goats in tropical and subtropical regions which causes high economic loss in the livestock industry. OBJECTIVES: The aim of this study was the differential detection of THEILERIA species in sheep using PCR method. METHODS: Two hundred blood samples of sheep were investigated in order to differentially diagnose THEILERIA species. DNA was extracted from blood samples and DNA samples were amplified using specific primers designed for 18S rRNA, TamS1 and TaSp genes. RESULTS: In this study, from 200 examined samples, 42 samples (21%) were infected by THEILERIA spp. and none of them were infected by Babesia spp. Moreover, from these 42 positive samples, 24 samples (57. 1%) were only infected by T. ovis. 12 samples (28. 5%) were only infected by T. lestoquardi, 2 samples (4. 7%) were only infected by T. ANNULATA and 4 samples (9. 5%) were simultaneously infected by T. lestoquardi and T. ovis. The results of nucleotide sequencing showed that PCR product of 18S rRNA from T. lestoquardi has 99 and 95% similarity with T. ANNULATA and T. ovis respectively. T. lestoquardi and T. ANNULATA showed 86% similarity. Also TaSp gene of T. ovis in comparison with T. ANNULATA and T. lestoquardi showed 96 and 86% similarity, respectively. CONCLUSIONS: In the present study could be shown that the two genes (TamS1 and TaSp) from examined three genes could be used for THEILERIA species specific diagnosis by PCR.

In order to effect of THEILERIA ANNULATA on some blood serum electrolytes' (Na, K, Ca, P, Cl, Fe , Zn, Mg) and non-electrolytes (urea, BUN, uric acid, creatinine) parameters, blood samples were taken from jugular vein of 50 males and females crossbred cattles naturally infected with THEILERIA ANNULATA in Ahwaz city and compared with same electrolytes and non electrolytes of blood serum in 68 males and females clinically healthy crossbred cattles in similar age groups. Statistically analysis show that, regardless of age and sex, cattles infected with THEILERIA ANNULATA have significantly higher serum urea, blood urea nitrogen and creatinine concentration and significantly lower calcium, phosphorus, magnesium, potassium, iron and cloride concentration than healthy cattle. There were not significant different between infected and uninfected cattles for uric acid, glucose, sodium and zinc (p>0.05). In> 6 and <24 month age groups of infected cattle, the mean values of uric acid were significantly lower and higher than healthy cattles, respectively (p<0.01). Glucose concentration in> 6 and 12-24 month age groups of infected cattles were lower than the uninfected cattles (p<0.05). In 12-24 and < 24 month age groups of infected cattle, the mean value of sodium was significantly lower than the healthy cattles (p<0.05).

The aim of this study was to detect and differentiate THEILERIA ANNULATA and T. lestoquardi (hirci) by PCR. Members of the genus THEILERIA are tick-borne hemoprotozoan parasites; those cause fatal and enervating diseases of cattle and sheep in Iran. In order to develop a specific method for detecting and identification of THEILERIA species, specific primers from the surface protein (SP) sequence were designed that allowed the specific diagnosis of T. ANNULATA and T. lestoquardi infection simultaneously. THEILERIA surface protein genes have consensus and variable sequences regions that allowed us to design the common primers for both species, which amplified two different PCR products. The results of this study demonstrated that a novel, simple, and high specific PCR for detecting and identifying T. ANNULATA and T. lestoquardi infection.

Background: Bovine theileriosis results from infection with obligate intracellular protozoa of the genus THEILERIA. The phylogenetic relationships between two isolates of THEILERIA ANNULATA, and 36 THEILERIA spp., as well as 6 out group including Babesia spp. and coccidian protozoa were analyzed using the 18S rRNA gene sequence.Methods: The target DNA segment was amplified by PCR. The PCR product was used for direct sequencing. The length of the 18S rRNA gene of all THEILERIA spp. involved in this study was around 1, 400 bp.Results: A phylogenetic tree was inferred based on the 18S rRNA gene sequence of the Iran and Iraq isolates, and other species of THEILERIA available in Gen Bank. In the constructed tree, THEILERIA ANNULATA (Iran vaccine strain) was closely related to other T. ANNULATA from Europe, Asia, as well as T.lestoquardi, T. parva and T. taurotragi all in one clade.Conclusion: Phylogenetic analyses based on small subunit ribosomal RNA gene suggested that the percent identity of the sequence of Iran vaccine strain was completely the same as Iraq sequence (100% identical), but the similarity of Iran vaccine strain with other T. ANNULATA reported from China, Spain and Italy determined the 97.9 to 99.9% identity.

THEILERIA are important hemoprotozoan parasites of cattle, transmitted by ixodid ticks leading to diseases which range from mild in apparent reactions to highly fatal diseases. This study aimed to determine the variety of THEILERIA species among cattle of Shahreza city, the south part of Isfahan Province. A total of 253 blood samples were randomly collected via the ear vein and jugular vein from healthy cattle for microscopy and molecular examination, respectively. The extracted DNA from blood cells was amplified by THEILERIA-all primers, which amplify an approximately 400bp DNA fragment from the region of the 18S rRNA gene from various members of the genus THEILERIA. All cattle positive samples were further analysed for the presence of THEILERIA ANNULATA and THEILERIA orientalis by specific semi-nested PCR. T. ANNULATA and T. orientalis were identified by specific semi-nested PCR in 54. 9% and 47% of cattle blood samples, respectively. Chi-square (x2) tests were used to compare molecular prevalence values relative to Season, Farm, Type, Hygiene, Vectors, Age, and Milk Yield. T. orientalis prevalence was significantly higher in cattle in summer as compared to those in other seasons (p < 0. 05). The prevalence rates of T. orientalis in different seasons, hygiene levels, and vectors were significantly different (p < 0. 05). T. orientalis prevalence was higher in spring, farms with tick vectors and normal level of Hygiene. The examination of 50 microscopic fields showed 14. 39% sensitivity and 99. 12% specificity compared to molecular examination. The Kappa coefficient between molecular and microscopy (50 fields) techniques indicated a poor level of agreement (Kappa= 0. 124). This study is the first molecular detection of THEILERIA species from cattle in the south of Isfahan province, Iran.

Background: Because of the strong immunologic responses of surface protein TaSp in THEILERIA ANNULATA infected host, we tried to characterize this protein in a T. ANNULATA isolate from Iran.Methods: The RNA prepared from T. ANNULATA infected cells was used to produce SMART-DScDNA.The Double strand cDNA was then amplified with primers derived from TaSp mRNA sequences.The PCR product was cloned in pTZ57R/T vector, sequenced and registered under accession no.JQ003240 in GenBank.Results: The sequence analysis showed 90%-94% nucleotide sequence identity and 68%-94% amino acid homology to the corresponding sequences of TaSp gene by T. ANNULATA, T. sp. china I, T. sp. china and T. lestoquardi and three T. ANNULATA reported from Iran respectively. Interestingly, the sequence analysis also showed small nucleotide sequence region near the 5` end in which the presented TaSp protein differed very strongly from the other known TaSp sequences. For the preparation of the recombinant protein, the cDNA was cloned in pQE-32 vector, the recombinant protein was prepared and assayed by THEILERIA infected bovine serum.Conclusion: The polymorphism in TaSp gene could be detected in intra-as well as inter species. The different characterized TaSp proteins had a common identic region, which may be helpful for development of broad band vaccine based on the recombinant proteins. The polymorphism in this gene, make this protein also interesting for the diagnostic purposes.

THEILERIA ANNULATAis one of the protozoan parasites which is transmitted by hard ticks and causes tropical theileriosis in cattle and buffaloes. This disease is one of the most important economic loss producing causes in several countries. This survey was conducted on 90, 2-5 years old native cattle (30 healthy, 30 involved with theileriosis and 30 asymptomatic cattle infected with THEILERIA). Blood samples were collected and then some hematologic and biochemical parameters were analyzed. The results revealed that in the involved group, RBC, PCV, HB, MCHC and WBC measures were significantly reduced compared with control group (P<0.05). The number of eosinophils in the involved and infected asymptomatic group were significantly higher than control (P<0.05), but the number of neutrophils were significantly less than control and asymptomatic groups (P<0.05). The counts of lymphocytes were also significantly lower in the involved and infected asymptomatic cattle than controls (P<0.05). Only PLT, lymphocyte, eosinophil and neutrophil rates had significant changes in asymptomatic infected cattle (P<0.05). The serum activity of ALT, AST, ALP and the amount of BUN were also significantly higher in the involved group compare to controls (P<0.05). The present investigation showed that unlike the involved group, studied parameters were returned to normal range in asymptomatic infected cattle after recovering from the disease.

Theileriosis is one of the most important protozoan parasitic diseases in Iran that causes economic losses and mortality in livestock. The disease is transmitted by Ixodid ticks and is characterized by fever, anemia, anoxia, and eventually death. Diagnosis of the disease was previously based on staining of blood and lymph nodes samples by Giemsa method. The use of molecular methods is preferred over previous methods due to their high sensitivity and specificity. Due to the fact that Ixodid ticks that can carry the disease are found in many traditional Iranian farms, it is very important to identify and introduce these ticks. The aim of this study was to molecularly identify THEILERIA ANNULATA infection in ticks isolated from cows in Urmia region. A total of 100 samples from herds with a history of THEILERIA ANNULATA infection in the area were collected and transferred to the laboratory. Ticks were identified using diagnostic keys. Salivary glands of ticks were separated and analyzed by PCR after isolation using THEILERIA ANNULATA specific primer (N516, N517). Out of 100 ticks, ten genera and species including Hyalomma anatolicum anatolicum (33%), Hy. detritum (16%), Boophylus anulatus (16%), Dermacentor marginatum (14%), Rhipicephalus bursa (9%), Hy. anantolicum excavatum (7%), D. niveus (4%) and Haemaphysalis punctate (1%). The predominant tick was identified as Hyalomma anatolicum anatolicum. PCR test showed the presence of 721 bp specific band on gel in 62% of tick samples. The high rate of infection in vector mites in the area indicates their high potential for infection of cows with the agent of THEILERIA ANNULATA.