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.

BACKGROUND: Babesiosis and Thosis are parasitic tick-borne diseases that cause a lot of economic loss in livestock Industry. Objectives: The purpose of the present study was to detect Babesia and Theileria infection in goats and and vector ticks in goats in Mashhad. Methods: One hundred blood samples of goats and 246 ticks were collected from some suspected flocks with history of piroplasmosis. The samples were transported to laboratory under cold condition. Blood smears were prepared and stained by Geimsa method and examined with a light microscope at ×1000 magnitude. The collected ticks were separated into tick pools of five according to their species and sex. The blood, salivary gland and ovaries of tick samples were examined using specific primers of Babesia.spp and Theileria.spp by semi nested-PCR. Results: Piroplasm bodies were not observed in any blood samples of goat in Mashhad. In a total of 246 collected ticks, seven species were identified as follows: R. turanicus 127(51.6%), D. marginatus 67 (27.2%), Hy. marginatum 44 (17.9%), R. sangunincus 4(1.6%), Hy. anatolicum 2(0.8%), Hy. asiaticum 1(0.4%) and Heam. sulcata 1(0.4%). Dominant tick species of goats in Mashhad suburb were R. turanicus and D. marginatus. The results of PCR showed that none of the blood samples were infected with Babesia spp. and Theileria spp. Also, Theileria infectoin was detected in a sample salivary glands  of Hy. marginatum. ConclusionS: Based on microscopic and molecular results, no  Theileria spp. and Babesia spp. infection were detected in goats. R.turanicus was the dominat tick species and Theileria spp. infection was detected in one sample of Hy.marginatum.

The objectives of the present study were to determine the infection rate of equine piroplasmosis (EP) in horses and ponies in Kelantan,Malaysia and compare the microscopic examination with competitive enzymelinked immunosorbent assay (cELISA) test as methods for diagnosis of EP. 306 blood samples were randomly collected from equids including 148 horses and 158 ponies in various districts of Kelantan, from September 2013 to March 2014. Based on microscopic examination of the staining blood smears, the infection rates ofTheileria equi, Babesia caballi and of both infections in horses were 19.59%, 25% and 8.78% respectively, whereas in ponies theinfection rates were 14.55%, 19.62%, and 5.69% respectively. Based on cELISA test, the infection rates of T. equi, B. caballi and of both infections in horses were 50.67%, 62.16% and 33.10% respectively,whereas in ponies, the infection rates were 51.89%, 63.92% and 35.44% respectively. No significant difference were observed between equids species associated with a seroprevalence of T. equi, B. caballi andof both infections (P≤ 0.05). According to the Kappa value there was no compatibility between microscopic examination and cELISA on the diagnosis of T. equi, B. caballi and of both infections which were 0.235, 0.013 and 0.080 respectively. In conclusion, the current results for this research work indicate that equine piroplasmosis is widespread in Kelantan, Malaysia and cELISA test is more efficientthan microscopic examination for diagnosis of EP.

Bovine haemoparasites are cosmopolitan in distribution and are known to cause substantial losses to the cattle industry. In spite of their economic importance, there remains a dearth of information on their molecular epidemiology in many parts of the world including Malaysia. To ascertain the molecular prevalence and species co-infection of bovine haemoparasites in the country, blood samples were collected from 1,045 heads of beef and dairy cattle on 43 farms from six geographical zones throughout Peninsular Malaysia. Samples subjected to PCR amplification of parasite species-specific genetic fragments revealed that Anaplasma marginale was the most prevalent haemoparasite (72.6%),followed by Theileria orientalis(49.8%),Candidatus Mycoplasma haemobos ( 47. 0 % ),Babesia bovis(32. 5%), Babesia bigemina (30.5%) and Trypanosomaevansi(17.9%). A high percentage (92.1%) of cattle was infected with either one or more haemoparasites. Triple haemoparasite species co-infection was the most prevalent (25.6%), followed closely by double species co-infection (25.1%). The most common (8.8%) and significantly correlated(rs= 0.250; p<0.01) combination was A. marginale+ T.orientalis. The present study constitutes the first attempt in the country to document the molecular prevalence and species co-infection of bovine haemoparasites over a wide spatial distribution. The data obtained will facilitate treatment, control and prevention measures to improve the local cattle industry.

A cDNA clone encoding a prohibitin-like protein (Toprh) was isolated from a piroplasm cDNA library of Theileria orientalis and its nucleotide sequence was determined. An open reading frame, encoding a polypeptide of 278 amino acid residues, was found in Toprh cDNA sequence. An intron of 89 bp was identified when this cDNA clone was compared with the Toprh gene in the genome of T. orientalis. The deduced amino acid sequence of Toprh shares 93.8, 93.1 and 69.1% identities with the prohibitins of T. parva (from chromosome 1), T. annulata (from chromosome 1), and Plasmodium falciparum, (from chromosome 10), respectively. By Western blot analysis, Toprh was found to be expressed in the piroplasm stage of the parasites.

The donkey population has remained unchanged in the last two decades despite a decrease in the overall population of equids, emphasizing the usefulness of the donkey as a draught and pack animal. Piroplasmosis in donkeys, caused by Theileria equi and Babesia caballi, has been recognized as a serious problem of major economic importance as the affected animals manifest decreased working capacity, loss of appetite, etc. In tropical countries, T. equi infections are more wide-spread and pathogenic than those caused by B. caballi. Donkeys usually remain asymptomatic carriers with positive antibody titres throughout life. Transmission of infection occurs from animal to animal through ticks such as Hyalomma spp. Rhipicephalus spp. and Dermacentor spp. The clinical form of the disease is diagnosed by peripheral blood smear examination, but in carrier donkeys it is very difficult to demonstrate the parasite in stained blood smears as the parasitaemia is extremely low. For diagnosis of such low grade infection or carrier animals, serological tests and DNA-based molecular diagnostic techniques, which are discussed in the present review, have become mandatory. Currently, there is no suitable pharmacotherapy available to clear the T. equi infection from affected donkeys, though some new drugs and drug combinations used against this disease condition have been discussed. In the present situation, there is an urgent need for international cooperation and coordination for development of sensitive molecular diagnostic tools and effective pharmacotherapies for curtailment of the disease condition. Hence, it is imperative to develop and exchange reagents and technology developed through human resource sharing in the interest of sustainability of donkey husbandry.

Objective: The effect of Artemisia herba-alba methanolic extract monotherapy and combination therapies on the in vitro growth of several Babesia and Theileria parasites in vitro and mice was investigated in this study. Materials and Methods: Fluorescence assay using SYBR Green I stain was used to evaluate the antibabesial efficacy inhibitory of A. herba-alba either in vitro or in vivo. Hematological parameters in the treated mice were analyzed using a Celltac MEK-6450 computerized hematology analyzer. Results: Artemisia herba-alba reduced the growth of Babesia bovis, Babesia bigemina, Babesia divergens, Theileria equi, and Babesia caballi in vitro in a dose-dependent manner. The in vitro inhibitory impact of A. herba-alba on B. divergens and B. caballi cultures was amplified when combined with either diminazene aceturate (DA). In B. microti-infected mice, a combination therapy consisting of A. herba-alba and a low DA dose inhibited B. microti growth significantly (p < 0.05) better than treatment with 25 mg kg−1 DA. Conclusions: These data show that A. herba-alba, when paired with a modest DA dose, could be a promising medicinal plant for babesiosis treatment. [J Adv Vet Anim Res 2022; 9(2.000): 267-274]