Theileriosis In Cattle Research Articles

Hematological profile and pathology of theileriosis in cattle from coastal districts of Odisha

Hematological profile and pathology of theileriosis in cattle from coastal districts of Odisha

Investigations on First Confirmed Outbreak of Ovine Theileriosis (Theileria luwenshuni) from Maharashtra State, India

Background: Clinical theileriosis of small ruminants is tick-borne disease caused by Theileria lestoquardi, Theileria uilenbergi and Theileria luwenshuni. Theileria annulata, the causative agent of bovine tropical theileriosis in cattle, can also infect sheep but does not cause any significant illness. It is one of the economically important diseases. There are no reports of ovine clinical theileriosis from Maharashtra state and there is paucity of information on its epidemiology. This paper reports first confirmed outbreak of ovine theileriosis based on clinical signs, microscopic examination, PCR and sequencing in the Maharashtra State of India. Methods: Whole blood samples from 22 ailing sheep were collected and subjected to hematological examination. Blood smears stained with Leishman’s stain were examined under 100X objective of the microscope. The blood samples from sheep found positive by microscopic method were subjected to PCR detection of 18S rRNA gene of hemoprotozoa and then for nucleotide sequencing and sequence analysis.Conclusion: Samples from 14 out of 22 sheep were found positive for piroplasms of Theileria spp by light microscopy. All positive samples were further confirmed by PCR detection of 18S rRNA gene of hemoprotozoa. PCR amplification yielded expected product of 1750 bp for all samples. BLAST and phylogenetic analysis of one sample revealed high sequence homology with T. luwenshuni reported from India and other countries. Characteristic clinical signs like fever, progressive anaemia, laboured breathing, lymphadenopathy, debility and non-responsiveness to antibiotic therapy were recorded. The animals responded to specific treatment against theileriosis. It is the first ever confirmed report of ovine theileriosis in Maharashtra state of India and hence reported.

Cattle Theileriosis: Prevalence and Comparative by Using ELISA and Blood Smears Techniques in Sulaimani Province - Kurdistan Region / Iraq.

Isolated one hundred fifty adult female cattle (local breed) one hundred infested with a hard tick (Ixodidae) and show some clinical signs for a search of theileriosis such as; enlargement pre-scapular lymph node and pale of the eye mucus membrane, fifty of them use a control (Non infested cattle) during the period of April to November 2018 in Sulaimani province. Indicated the highest sensitivity (prevalence) of Theileria annulata when examined by ELISA technique 63% (63/100), the prevalence rate of macroscopically detection to T. annulata merozoite was 48% as the following, in mixed lymph node biopsy cells and blood smears technique was 22%, the blood and lymph node biopsy smear examinations were 15% and 11% respectively. Distinguished the total prevalence rate in control cattle was 6% and 4% in the ELISA technique and microscopically method respectively. Sensitivity values and positive predictive noted a highly in ELISA serolog-ical test 63% and 95.45% respectively, while in mixed lymph node biopsy and blood smears examined 22% and 100% respectively. During the study collected 1368 hard tick and identified to three genera in infested cattle of Ixodidae; Hyalomma species are the highest number percentage (44.8%), followed Boophilus species (28.6%) and the Rhipicephalus species is the lowest percentage (26.6%). This study conducted to find out the prevalence the local cattle with theileriosis (T. annulata) by the microscopic examination for both blood and lymph biopsy smear (merozoite) and compared with the (ELISA) technique.

Determination of the Infectivity of Cryopreserved Theileria annu-lata Sporozoites in Tick Derived Stabilates Iran Ak-93 Strain, by In Vivo and In Vitro Methods

The protozoan parasite Theileria annulata is the causative agent of tropical theileriosis in cattle. Vaccination is recommended by administration of attenuated schizont-infected cell lines. The expected protective immunity post-vaccination can be demonstrated by challenge test through inoculation of highly virulent infective sporozoites. The aim of this study was to produce Hyalomma anatolicum anatolicum tick infected with T. annulata (local strain) for preparation of tick-derived sporozoite stabilates for molecular characterization and infectivity test assay. A local T. annulata strain was used for experimental infection of calves. A field isolate of H. a. anatolicum was isolated, laboratory-reared and infected by blood-feeding on Theileria infected above-mentioned calves. The infectivity of calf, tick and prepared stabilate were confirmed by clinical signs of theileriosis, microscopic inspection, RT-PCR and in vitro cell culture. The tick stabilate was prepared and cryopreserved in liquid nitrogen. The infectivity of the tick stabilate was verified by in vivo bioassay, in vitro cell culture infection, microscopic inspection in salivary glands and RT-PCR assay. The in vitro produced cell line in this study was characterized by T. annulata Cytochrome b gene analyzing. The infectivity of a new prepared tick-derived sporozoite stabilate was confirmed in susceptible calves; by microscopically, post mortem, tick microscopic and molecular assays. Moreover, naïve PBMCs were transformed and proliferated by T. annulata infected tick stabilate to immortal T. annulata schizont infected cell line. The potent infective sporozoite tick derived stabilate could be used for vaccine efficacy and challenge test as well as in vaccine development.

Efficacy of Arteether Against Theileriosis in Cattle

Efficacy of Arteether Against Theileriosis in Cattle

Discovery of a new Theileria sp. closely related to Theileria annulata in cattle from Sri Lanka

Theileria annulata is a haemoprotozoan parasite that causes a cancer-like illness known as tropical theileriosis in cattle. In the course of analyzing the genetic diversity of T. annulata in Sri Lanka, we observed that merozoite-piroplasm surface antigen (tams1) and surface protein (tasp)-like gene sequences obtained from bovine blood DNA samples, which were PCR-positive for T. annulata, were conserved but shared low identity with T. annulata GenBank sequences. Moreover, the 18S rRNA sequences from the Sri Lankan samples contained ten unique single-nucleotide polymorphisms compared with all known T. annulata sequences. The cytochrome b (cob) gene sequences isolated from the Sri Lankan samples were highly conserved and shared low identity scores with similarly conserved T. annulata sequences from GenBank. Phylogenetic analysis showed that the Sri Lankan tams1-like, tasp-like, 18S rRNA, and cob sequences clustered together and formed sister clades to the common ancestors of all known T. annulata and Theileria lestoquardi sequences. These findings demonstrated that the Sri Lankan cattle were not infected with T. annulata but with a new Theileria sp. (designated as Theileria sp. Yokoyama) closely related to T. annulata.

Chemotherapy against piroplasmosis and anaplasmosis in cattle

It has been established that Teilercid is a specific medication in the treatment of cattle theileriosis. Two-time intramuscular injection of Teilercid in a dose of 1 ml per 20 kg of live weight with an interval of 48 hours in combination with symptomatic and pathogenic agents has a high therapeutic effect against cattle theileriosis. Babesan is established to have high therapeutic efficacy in babesiosis (Babesia bigemina) and cattle anaplasmosis (Anaplasma marginale) at a dose of 2.0 and 3.0 ml per 100 kg of live weight, respectively, injected intramuscularly in combination with symptomatic and pathogenic agents. Neozidinum M at a dose of 1 ml per 20 kg of live weight intramuscularly has the efficiency of 93-100% in the treatment of cattle babesiosis and theileriosis.

Molecular characterization of Theileria spp. in livestock and the first report on the occurrence of Theileria sp. OT3 in Iran.

Theileria lestoquardi, T. ovis, and T. annulata are recognized as major causative agents for ovine and cattle theileriosis in Iran, respectively. Recently, there have been reports on the presence of Theileria spp. (Theileria sp. OT1, Theileria sp. OT3, and Theileria sp.). In this study, 37 blood samples were collected from sheep and cattle with clinically suspected Theileria infection in the Northwest of Iran. The samples were analyzed using a light microscope. DNA samples were amplified via nested-polymerase chain reaction (PCR) of 18S rRNA gene. The amplicons were digested with HpaII, following restriction fragment length polymorphism (RFLP) and sequenced to reconfirm Theileria species. The microscopic examination indicated that 4 out of 37 (10.8%) blood samples were infected with Theileria spp. Based on the nested PCR-RFLP and sequencing data, 5.4%, 13.5%, and 27% of blood samples were infected with Theileria sp. OT3, T. ovis, and T. annulata, respectively. The pairwise distance matrix of Theileria sp. OT3 showed 99.8-100% identity and 0-0.2% divergence in comparison with the registered sequences. The present study is the first report of Theileria sp. OT3 in Iran. To the evaluate evolution of Theileria spp. and providing resultant genetic data, further research with a larger sample is necessary in the region.

In vitro treatment of Besnoitia besnoiti with the naphto-quinone buparvaquone results in marked inhibition of tachyzoite proliferation, mitochondrial alterations and rapid adaptation of tachyzoites to increased drug concentrations.

We here assessed the in vitro efficacy of the naptho-quinone buparvaquone (BPQ) against Besnoitia besnoiti tachyzoites in vitro. BPQ is currently licensed for the treatment of theileriosis in cattle in many countries, but not in the EU. In 4-day treatment assays, BPQ massively impaired tachyzoite proliferation with an IC50 of 10 ± 3 nm, and virtually complete inhibition was obtained in the presence of nm BPQ. Exposure to 1 µm BPQ leads to ultrastructural changes affecting initially the mitochondrial matrix and the cristae. After 96 h, most parasites were largely distorted, filled with cytoplasmic amylopectin granules and vacuoles containing components of unknown composition. Host cell mitochondria did not appear to be notably affected by the drug. However, upon prolonged exposure (14-16 days) to increased BPQ concentrations, B. besnoiti tachyzoites exhibited the capacity to adapt, and they resumed proliferation at dosages of up to 10 µm BPQ, albeit at a lower rate. These BPQ-adapted parasites maintained this lower susceptibility to BPQ treatment after freeze-thawing, and inspection by the transmission electron microscopy revealed that they underwent proliferation in the absence of structurally intact mitochondria.

Relationship between cardiac injury, selected biochemical parameters, DIC, and hemogram levels in cattle with theileriosis

This study was undertaken to investigate the effects of theileriosis on the cardiovascular system in cattle. Blood samples were collected from 50 cattle naturally infected with Theileria spp. (before and after treatment) as well as from 30 non-infected cattle. Hematological parameters and the serum concentrations of total cardiac troponin I (cTnI), aspartate aminotransferase (AST), creatine kinase (CK), and creatine kinase MB (CK-MB) were measured. Also coagulation parameters, plasma levels of activated partial thrombo plastin time (APTT), prothrombin time (PT) and D-dimer were measured. The presence of anemia and the levels of cTnI, CK-MB, CK and AST indicated statistically significant differences between infected and healthy cattle, but no statistically significant differences between the two groups were found in DD levels at the end of the study. Reduction in anemia and reduced levels of cTnI, CK-MB, CK, AST, APTT and PT were also found in post-treatment analyses. The results indicate that anemia, coagulation disorders and hypoxia in theileriosis in cattle can induce myocardial tissue damage.

Comprehensive structural analysis of the open and closed conformations of Theileria annulata enolase by molecular modelling and docking

Theileria annulata is an apicomplexan parasite which is responsible for tropical theileriosis in cattle. Due to resistance of T. annulata against commonly used antitheilerial drug, new drug candidates should be identified urgently. Enolase might be a druggable protein candidate which has an important role in glycolysis, and could also be related to several cellular functions as a moonlight protein. In this study; we have described three-dimensional models of open and closed conformations of T. annulata enolase by homology modeling method for the first time with the comprehensive domain, active site and docking analyses. Our results show that the enolase has similar folding patterns within enolase superfamily with conserved catalytic loops and active site residues. We have described specific insertions, possible plasminogen binding sites, electrostatic potential surfaces and positively charged pockets as druggable regions in T. annulata enolase.

Biochemical and in silico Characterization of Recombinant L-Lactate Dehydrogenase of Theileria annulata.

Theileria annulata is a parasite that causes theileriosis in cattle. Reports about drug resistance made essential to develop new drug. LDH of Theileria schizonts is the vital enzyme for its anaerobic metabolism. TaLDH gene was first cloned into pGEM-T cloning vector with two introns in our previous study. Here we report cloning of TaLDH without introns into pLATE 31 vector in E. coli BL21(DE3). Protein was in an inactive form. Two mutations were fixed to express the active protein. Protein was purified by affinity chromatography and evaluated by SDS-PAGE and size exclusion chromatography. Optimum pH of enzyme was performed in pH 7.5, and enzyme was stabilized at 20-40 °C. Enzyme kinetics of recombinant TaLDH were found to be in the direction of pyruvate to lactate K m 0.1324 and K i 4.295 mM, k cat, 44.55/s and k cat /K m, 3.3693 × 10(5)/M/s. 3D structure of TaLDH was predicted, and possible drug binding sites were determined by homology modelling.

Repurposing of antiparasitic drugs: the hydroxy-naphthoquinone buparvaquone inhibits vertical transmission in the pregnant neosporosis mouse model.

The three anti-malarial drugs artemiside, artemisone, and mefloquine, and the naphthoquinone buparvaquone known to be active against theileriosis in cattle and Leishmania infections in rodents, were assessed for activity against Neospora caninum infection. All four compounds inhibited the proliferation of N. caninum tachyzoites in vitro with IC50 in the sub-micromolar range, but artemisone and buparvaquone were most effective (IC50 = 3 and 4.9 nM, respectively). However, in a neosporosis mouse model for cerebral infection comprising Balb/c mice experimentally infected with the virulent isolate Nc-Spain7, the three anti-malarial compounds failed to exhibit any activity, since treatment did not reduce the parasite burden in brains and lungs compared to untreated controls. Thus, these compounds were not further evaluated in pregnant mice. On the other hand, buparvaquone, shown earlier to be effective in reducing the parasite load in the lungs in an acute neosporosis disease model, was further assessed in the pregnant mouse model. Buparvaquone efficiently inhibited vertical transmission in Balb/c mice experimentally infected at day 7 of pregnancy, reduced clinical signs in the pups, but had no effect on cerebral infection in the dams. This demonstrates proof-of-concept that drug repurposing may lead to the discovery of an effective compound against neosporosis that can protect offspring from vertical transmission and disease.

Prevalence of Tropical Theileriosis in Cattle in Chhattisgarh State

Prevalence of Tropical Theileriosis in Cattle in Chhattisgarh State

Emergence of oriental theileriosis in cattle and its transmission through Rhipicephalus (Boophilus) microplus in Assam, India.

Aim:The aim of the present study was to investigate the presence of Theileria in blood samples of crossbred and indigenous adult cows raised under unorganized small scale farming system in a Babesia and Anaplasma endemic geographical area from Assam, India and to see its transmission through Rhipicephalus (Boophilus) microplus ticks.Materials and Methods:For the present study, 57 clinical cases of cattle suspected to be of hemoparasitic infections were taken into consideration. The parasites were identified based on morphology in giemsa stained blood smear followed by polymerase chain reaction (PCR). Sera samples were tested for T. annulata antibodies in plate and Dot-ELISA. PCR was also conducted in eggs of Rhipicephalus (Boophilus) microplus tick collected from a Theileria orientalis positive animal.Results:PCR amplified 1124, 776, and 160 bp DNA fragments of B. bigemina (64.91%), T. orientalis (21.05%) and A. marginale (14.03%), respectively. This assay further conducted in 12 T. orientalis positive blood samples with primers of Buffeli, Chitose, and Ikeda variants of T. orientalis showed 3 samples positive to Ikeda type and none for Buffeli and Chitose. Babesia bovis and Theileria annulata specific primers also did not amplify any fragment during the PCR assay of the blood samples. Further, all sera samples tested negative to T. annulata antibodies in Plate and Dot-ELISA. PCR conducted in eggs of R (B).microplus tick collected from a T. orientalis positive animal revealed presence of the parasite DNA. Gradual improvement in physical condition leading to complete recovery in 10 out of 12 T. orientalis infected clinical cases treated with buparvaquone(at 2.5mg/kg.b.wt I/M) was the feedback obtained from field veterinarians and the cattle owners.Conclusion:The present investigation represents the first report of occurrence of T. orientalis in cattle of Assam with involvement of pathogenic Ikeda strain in clinical outbreaks and its possible natural transmission by R (B). microplus through the transovarian mode.

An additional records of Hyalomma marginatum rufipes Koch, 1844 (Acari: Ixodidae) in southwestern and southern Iran with a molecular evidence

Hyalomma marginatum rufipes (Acari: Ixodidae) along with three closely related subspecies is considered as marginatum group. The subspecies had proven as main vector of Crimean Congo hemorrhagic fever virus, the cause of human death in Asia, Africa and Europe. This tick is also a vector of parasitic protozoan Theileria annulata, agent of tropical theileriosis in cattle. Nonetheless, taxonomical status of this tick not recognized or confirmed in tick’s fauna of Iran, then we decided to show most taxonomic characteristics and confirm the presence of this subspecies in Iran by molecular methods. Tick specimens were collected from cattle in Manujan township, Kerman province, southern Iran. Specimens were identified morphologically using suitable taxonomical identification keys. The morphologically identified specimens were subjected to molecular studies. Morphological and COI gene analysis clearly confirmed the occurrence of H. m. rufipes in Iran, however, according to ITS2 fragment H. m. rufipes can be the same H. m. marginatum. Thus, it seems that based on most popular molecular markers, H. m. rufipes and its relative H. m. marginatum really should be assigned as a polymorphic species H. marginatum.

Investigating the first outbreak of oriental theileriosis in cattle in South Australia using multiplexed tandem PCR (MT-PCR)

This study investigated the first outbreak of oriental theileriosis in a herd of beef cattle in South Australia using a newly established multiplexed tandem PCR (MT-PCR) to identify, differentiate and quantitate the four genotypes (buffeli, chitose, ikeda and type 5) of Theileria orientalis recognised to occur in Australasia. Following clinical diagnosis of oriental theileriosis (based on clinical signs, laboratory findings and post mortem examination), 155 blood samples were collected from individual cows (n=85) and calves (n=70), and tested by MT-PCR. In total, 117 (75.48%) cattle were shown to be test-positive for T. orientalis. All four genotypes were detected, and ikeda had the highest prevalence (90.6%; 106/117), followed by buffeli (83.8%; 98/117), chitose (18.8%; 22/117) and type 5 (5.1%; 6/117). Mixed infections with genotypes buffeli and ikeda had a higher prevalence (55.5%; 65/117) than any other combination of genotypes. The prevalences of buffeli and ikeda were significantly higher (P<0.005) than those of chitose and type 5. The average intensity of infection with genotype ikeda (329,775 DNA copies) was significantly higher (P<0.0001) than buffeli (212,843) and chitose (125,462). This study reinforces the utility of MT-PCR as a diagnostic tool for rapidly investigating oriental theileriosis outbreaks in cattle herds and as a pre-movement screening test for preventing the introduction of this disease into non-endemic regions.

Correlation between Platelets and Red Blood Cells Parameters in Cattle Theileriosis

The purpose of the study is to investigate the values for platelets (PLT) count, plateletcrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW) and the correlation with the parallel red blood cell parameters, packed cell volume (PCV), mean corpuscular volume (MCV), and red blood cell distribution width (RDW) in cattle Theileriosis. A total number of 25 cows were subjected to study. Out of them 18 cows were suffered from Theileriosis, the remained 7 cows were clinically healthy and kept as control. Red blood cells (RBCs) and platelet indices were measured in whole blood samples collected on EDTA. Animals infected with Theileria were classified into two groups based on total erythrocytes count. Group 1: included animals that had total RBCs count above 5 x 106/mm3. Group 2: included animals that had total RBCs count below 5 x 106/mm3. Comparing data from theileria infected groups with the control group revealed that there was a significant decrease in total RBCs count only in group 2 (p&lt;0.01). MCV showed significant decreases in group 1 (p&lt;0.01) and group 2 (p&lt;0.05), PCV was significantly decreased in both groups (p&lt;0.01), RDW was significantly increased only in group 2 (p&lt;0.05), PLT count was significantly decreased only in group 2 (p&lt;0.05), PCT was significantly decreased only in group 2 (p&lt;0.05), no significant changes were observed in MPV and PDW. Comparing data from group 1 with group 2 revealed that RBCs count, PCV and PLT count were significantly higher in group 1 than group 2. Furthermore, MCV value was significantly higher in group 2 than group 1. RDW was significantly higher in group 2 than group 1. The present study revealed that in cattle Theileriosis, PLT indices greatly affected by the degree of anemia. Furthermore, the decrease in PLT count and PCT usually accompany oligocythemia.

Molecular prevalence and genetic diversity of bovine Theileria orientalis in Myanmar

Theileria orientalis is a causative agent of benign theileriosis in cattle and distributed in mainly Asian countries. In the present study, we examined the prevalence of T. orientalis infection by PCR based on the major piroplasm surface protein gene (MPSP) sequences in cattle in Myanmar, followed by phylogenetic analysis of the MPSP genes. The MPSP gene was amplified in 258 of 713 (36.2%) cattle blood DNA samples collected from five cities in different geographical regions of Myanmar. Phylogenetic analysis of MPSP sequences from 54 T. orientalis-positive DNA samples revealed the presence of six allelic genotypes, including Types 1, 3, 4, 5, 7, and N-3. Types 5 and 7 were the predominant types detected. Sequences of the MPSP genes detected in Myanmar were closely related to those from Thailand, Vietnam or Mongolia. These findings suggest that movement of animals carrying T. orientalis parasites between Southeast Asian countries could be a reason for the similar genotype distribution of the parasites in Myanmar.

Detection of theileriosis in cattle and buffaloes by polymerase chain reaction.

Bovine tropical theileriosis caused by Theileriaannulata is a tick-borne disease of great economic importance in tropical and subtropical regions of the world. The present study was undertaken to detect theilerosis in cattle and buffaloes by polymerase chain reaction (PCR). The diagnosis of theileriosis is usually carried out by blood smear staining technique, which is not sufficiently sensitive to detect the piroplasms in the carrier animals. In this study, a total of 116 samples were collected from infected as well as apparently healthy cattle and buffaloes. Screening of blood smears by Giemsa staining detected 15 samples (12.93%) positive for Theileria piroplasms out of 116 samples. However, the PCR based screening using the specific primers from the major merozoite-piroplasm surface antigen sequence of T.annulata (Tams1) gene detected 74 samples (63.79%) positive for T.annulata which included 59 samples found negative by Giemsa staining. Our study suggests that the PCR based screening is more sensitive and accurate method for diagnosis of tropical theileriosis in cattle and buffaloes.