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Purpose: Tropical theileriosis is a tick-borne haemoprotozoan disease, and cardiac function assessment in buffaloes with theileriosis was poorly documented. Methods: The Present study was carried out from April 2022 to December 2022. Theileriosis was confirmed by microscopic examination of stained blood smears and lymphnode smears further confirmed by PCR assay. Electrocardiography was performed by using the base apex lead system, and echocardiography was performed by using the right parasternal view. Results: The incidence of theileriosis was 16.25% by examination of stained blood smears, and 30.42% by PCR examination in 240 buffaloes. Repeatedly noted clinical signs were the absence of rumination, anorexia, loss of milk yield, depressed demeanour, emaciation, hyperthermia, lymphadenopathy, tick infestation, tachycardia, cardiac arrhythmia, and increased intensity of heartbeat. Haematological findings disclosed decreased haemoglobin, packed cell volume, total erythrocyte count, and neutrophils; increased eosinophils and monocytes. Serum biochemical findings revealed decreased albumin, albumin/globulin ratio, glucose, calcium, phosphorous, sodium, potassium, and chloride; increased globulin, aspartate aminotransferase, bilirubin, blood urea nitrogen, creatinine, cholesterol, lactate dehydrogenase, gamma-glutamyl transferase, and creatine kinase myocardial band isoenzymes. Electrocardiography explorations were sinus tachycardia, broad T wave, and sinus arrhythmia. Echocardiography examination showed ventricular wall thickening, cardiac chamber dilatation, valvular defects/valvular regurgitation, and pericarditis/cardiac tamponade. Conclusion: The present research proposes the changes in the electrocardiography and echocardiography findings in buffaloes with theileriosis, which are essential in clinics to identify the secondary complications during theileriosis and formulate therapeutics. [ABSTRACT FROM AUTHOR]

Background:Theileria spp. are responsible for ovine and caprine theileriosis, leading to significant morbidity and mortality in small ruminants. The present study aims to investigate Theileria spp. infections in small ruminants from Southern Punjab in Pakistan, and genetic characterize revealed Theileria spp. isolates. Methods: A total of 93 sheep and 107 goats were sampled between May and August 2022. Blood smears were examined microscopically, and PCR amplification targeting the 18S rRNA gene was performed to detect Theileria spp. Additionally, specific PCR assays targeting 18S rRNA and ms1 partial sequences were used to identify Theileria ovis and T. lestoquardi, respectively.  Results: The prevalence of Theileria spp. was significantly higher using PCR (13.5%) compared to microscopic screening (5%). Sheep showed a higher prevalence rate (19.4%) compared to goats (8.4%) (p = 0.024). Young sheep aged ≤ 1 year were more commonly infected with Theileria spp. (41%) compared to older sheep (p = 0.006). The prevalence of Theileria spp. was higher in sheep-only herds (37.3%) compared to goat-only herds (18%) or mixed-species herds (8.1%) (p = 0.015). The prevalence rates of T. ovis and T. lestoquardi were 9% and 2.5%, respectively, with four animals (2 goats and 2 sheep) showing co-infection. Phylogenetic analysis revealed that our T. ovis 18S rRNA sequence clustered with previously reported sequences from sheep in Turkey, China, Spain, and goats in Tanzania. The obtained T. lestoquardi ms1 partial sequence formed a distinct cluster from other T. lestoquardi isolates in Pakistan and neighboring countries.  Conclusion:Theileria spp. co-circulation in Pakistani small ruminants, particularly the presence of T. ovis and T. lestoquardi, highlights the need for attention from animal health decision-makers due to their financial and health impacts. [ABSTRACT FROM AUTHOR]

Background: In Algeria, data on the epidemiology of theileriosis in small ruminants are limited. Objectives: The current study investigates the seroprevalence of Theileria spp. in goats from the M'Sila region, Central Algeria. Methods: Blood samples of 128 goats from 19 farms were collected from the locality of Maâdid. The indirect fluorescence antibody test (IFAT) was performed to test the goats' sera for antibodies against Theileria ovis and Theileria lestoquardi. Results: Out of 128 tested samples, 21 sera (16.40%) were positive for T. ovis antibodies. All samples were seronegative for T. lestoquardi. The seroprevalence of T. ovis varied from 10% to 30% on farms. The seropositivity rates did not differ significantly with age, sex, or breeding system of goats. Tick-infested goats were significantly more seropositive than no-infested goats. Conclusion: The present study reports essential data on the epidemiology of caprine theileriosis from Central Algeria. [ABSTRACT FROM AUTHOR]

Background & objectives: Theileriosis is an important tick-bome hemoprotozoan disease of cattle which causes severe economic loss due to morbidity and mortality. A diagnostic test having high sensitivity, specificity and easy application at the field level is the need of the hour. In this regard Loop-mediated isothermal amplification (LAMP) is proven to be a sensitive, easy and time efficient method. One of the major obstacles for the application of LAMP is the difficulty in maintaining the cold chain to preserve reagents. Thus, the challenge is to develop a LAMP kit in a ready-to-use format with dried reagents useful for quick and simple application in field conditions. Methods: The optimized reaction of wet LAMP was followed for the standardization of dry LAMP with certain modifications which are needful. The major modification is vitrification technology of enzyme using trehalose. Results: LAMP assay (dry and wet LAMP) was found to be more sensitive (100%) when compared to microscopy (69.5%) and PCR (86.9%). It was observed that the dry LAMP reaction tubes at room temperature as well as refrigeration temperature provided successful amplification till 7 weeks. Interpretation & conclusion: The drying conditions of LAMP reagents were optimized, and finally managed to dry them in a single reaction tube without reducing the sensitivity. This technology enables us to transport LAMP kits to areas where the cold chain is not easily available.

Theileriosis, a parasitic disease, caused by Theileria spp. and transmitted through ticks, poses a significant threat to livestock, leading to elevated morbidity and mortality rates. This study investigated the incidence trend of theileriosis in Kerala, India, over three years (2019–21). Notably, the research unveiled a substantial upsurge in bovine theileriosis cases within Kerala during this period, partly attributed to the state’s severe flooding and landslides in 2018, triggered by incessant monsoon rains. The present study envisaged pinpointing the risk factors underlying the prevalence of theileriosis in Kerala. Employing linear discriminant analysis, key environmental and remote sensing variables influencing the disease’s incidence were identified. Subsequently, these risk factors underwent climate disease modelling, leading to the creation of risk maps. To predict areas sensitive to theileriosis outbreaks in Kerala, two regression models and nine machine learning models were employed. The gradient boost and random forest models demonstrated the most accurate fit among these. The study also estimated the basic reproduction number (R0), which ranged from 0.89 to 1.8. This value indicates a high potential for Theileria spp. transmission within the study area. Consequently, the research outcomes offer valuable insights into pinpointing high risk theileriosis locations in livestock in Kerala. [ABSTRACT FROM AUTHOR]

Piroplasm including Babesia spp. and Theileria spp. in cattle can cause illness that affects livestock productivity, resulting in significant production losses, especially in tropical and subtropical regions such as Thailand. This study aimed to investigate the prevalence of bovine piroplasms and to identify these blood parasites based on the 18S ribosomal RNA gene in cattle in the northeastern part of Thailand. Piroplasmid infections among beef and dairy cattle were examined using nested PCR. Furthermore, amplicon DNA was sequenced and analyzed, and a phylogenetic tree was constructed to determine the genetic diversity and relationships of the parasite in each area. A total of 141 out of 215 (65.6%) cattle were positive for infection with Babesia or Theileria. DNA analysis revealed that infection by Babesia bigemina, Babesia bovis, Theileria orientalis, Theileria sinensis, and Theileria sp. were common piroplasms in cattle in this region, with a high sequence shared identity and similarity with each other and clustered with isolates from other countries. This study provides information on the molecular epidemiology and genetic identification of Babesia spp. and Theileria spp. in beef and dairy cattle to provide a better understanding of piroplasm infection in cattle in this region, which will help control these blood parasites. Moreover, this is the first report identifying T. sinensis circulating among Thai cattle. [ABSTRACT FROM AUTHOR]

Pathogens such as Plasmodium , Babesia , and Theileria invade and multiply within host red blood cells, leading to the pathological consequences of malaria, babesiosis, and theileriosis. Establishing continuous in vitro culture systems and suitable animal models is crucial for studying these pathogens. This review spotlights the Babesia duncani in culture-in mouse (ICIM) model as a promising resource for advancing research on the biology, pathogenicity, and virulence of intraerythrocytic parasites. The model offers practical benefits, encompassing well-defined culture conditions, ease of manipulation, and a well-annotated genome. Moreover, B. duncani serves as a surrogate system for drug discovery, facilitating the evaluation of new antiparasitic drugs in vitro and in animals, elucidating their modes of action, and uncovering potential resistance mechanisms. The B. duncani ICIM model thus emerges as a multifaceted tool with profound implications, promising advancements in our understanding of parasitic biology and shaping the development of future therapies. [ABSTRACT FROM AUTHOR]

This veterinary clinical and pathology case report describes a severe clinical case of theileriosis in adult cattle with prolonged recumbency and terminal wasting because of complications due to chronic theileriosis in two adult (2-year-old) Jersey-Friesian Cross. The case was referred to the large animal clinic of the University Veterinary Hospital (UVH), Faculty of Veterinary Medicine, Universiti Putra Malaysia (UPM). The animals were presented as recumbent, dull, depressed, hypothermic, dehydrated, and cachexic. There were poor responses of the patients to fluid therapy and non-steroidal anti-inflammatory treatments, and they both died within 24 hrs of clinical presentation. Haematology (complete blood count and serum biochemistry) results revealed haemoconcentration and faecal examinations were negative for liver fluke infestation and blood smear examination was positive for intraerythrocytic piroplasms of Theileria (0.01% parasitaemia). Post-mortem examination further revealed advanced cachexia, generalised serous atrophy of fat, hepatomegaly, hepatic congestion, pulmonary congestion, and pulmonary oedema. Histological findings revealed atrophy of the hepatic cord, hepatic congestion, splenic hemosiderosis, pulmonary oedema and congestion. The clinicopathological findings supported an aetiological diagnosis of a severe clinical case of Theileriosis with terminal wasting because of complications due to chronic Theileriosis infection. [ABSTRACT FROM AUTHOR]

Theileriosis is an important disease in exotic and cross bred dairy cattle in India. Group I was treated with therapeutic drug injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + Yakrifit liquid @ 50 mL twice daily for 7 days. Group II was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + Yakrifit boli @ 2 boli twice daily for 7 days, Group III was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + AV/SYL/12 @ 50 mL twice daily for 7 days, Group IV was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + AV/SYB/12 @ 2 boli twice daily for 7 days and Group V was treated with injection buparvaquone @ 2.5 mg/kg B.W. deep intramuscularly + Liv 52 bolus @ 2 boli twice daily for 7 days. In present study, all theileriosis infected groups showed 100 % recovery after treatment with standard drug buparvaquone and adjunct therapy (Yakrifit liquid, Yakrifit boli, AV/SYL/12 liquid, AV/SYL/12 boli and Liv 52 bolus) within 7 days. Within a week, all six animals showed clinical recovery as evaluated on the basis of clinical parameters which returned to normal and improvement in the hematological and biochemical parameters. [ABSTRACT FROM AUTHOR]

Theileriosis is the most important common vector (Tick) born and hemoprotozoan diseases (TBDS) which pose a serious threat to the livestock population in terms of mortality, reduced milk yield and lowered draft power. A total 1900 cases were suspected for theileriosis on the basis of clinical signs and out of these a total of 950 cases were found positive for theileriosis on the basis of blood smear examination. As per the stages of pregnancy and lactation, the cases were classified as cows in early pregnancy, cows in late pregnancy, cows in post parturient stage, cows in early and late lactation period. The cows in post parturient period were more susceptible to theileriosis. The incidence was higher in 2-6 years of age group, post monsoon period and mostly in cross bred animals. In most of the cases the clinical signs observed were reduced appetite, non-remittent fever, lacrimation and salivation and infrequent signs like swollen lymphnodes, respiratory symptoms and reduced milk yield. Haematological values of PCV, Hb, MCH and MCHC were significantly low in post parturient cows, followed by cows in late pregnancy, in early lactation, early pregnancy and late lactation period. [ABSTRACT FROM AUTHOR]

The Arabian Peninsula's endemic ungulate, Oryx leucoryx, was on the verge of extinction at the end of the 1970s. Despite the different reintroduction programs, the International Union for Conservation of Nature is still classifying it as Vulnerable. Among other factors, their vulnerability lies in their susceptibility to specific etiological agents that affect livestock, necessitating health monitoring and strict preventive/biosecurity measures. Within this frame, the current work investigated the determination of the etiological agent potentially involved with cutaneous lesions observed in eight males of Arabian oryx within one of the several national governance conservation programs. Microscopic examination from one animal specimen suggested theileriosis association, which was confirmed by molecular tools using 18S gene sequencing and the report of a novel Theileria sp. not clustering with previously reported antelope sequences. This finding prompts further explorations into the disease dynamics within the Arabian oryx population, especially with the scarcity of data in Qatar about tick-borne pathogens and their transmission. [ABSTRACT FROM AUTHOR]

Tropical theileriosis is a fatal leukemic-like disease of cattle caused by the tick-transmitted protozoan parasite Theileria annulata. The economics of cattle meat and milk production is severely affected by theileriosis in endemic areas. The hydroxynaphtoquinone buparvaquone (BPQ) is the only available drug currently used to treat clinical theileriosis, whilst BPQ resistance is emerging and spreading in endemic areas. Here, we chronically exposed T. annulata-transformed macrophages in vitro to BPQ and monitored the emergence of drug-resistant parasites. Surviving parasites revealed a significant increase in BPQ IC50 compared to the wild type parasites. Drug resistant parasites from two independent cloned lines had an identical single mutation, M128I, in the gene coding for T. annulata cytochrome B (Tacytb). This in vitro generated mutation has not been reported in resistant field isolates previously, but is reminiscent of the methionine to isoleucine mutation in atovaquone-resistant Plasmodium and Babesia. The M128I mutation did not appear to exert any deleterious effect on parasite fitness (proliferation and differentiation to merozoites). To gain insight into whether drug-resistance could have resulted from altered drug binding to TaCytB we generated in silico a 3D-model of wild type TaCytB and docked BPQ to the predicted 3D-structure. Potential binding sites cluster in four areas of the protein structure including the Q01 site. The bound drug in the Q01 site is expected to pack against an alpha helix, which included M128, suggesting that the change in amino acid in this position may alter drug-binding. The in vitro generated BPQ resistant T. annulata is a useful tool to determine the contribution of the various predicted docking sites to BPQ resistance and will also allow testing novel drugs against theileriosis for their potential to overcome BPQ resistance. [ABSTRACT FROM AUTHOR]

The hemostatic mechanism is crucial to stop bleeding while ensuring that tissues receive enough blood flow. This investigation examined selected haematological and coagulation parameters in different degrees of anemic Theileriosis infected cattle and study correlation between RBCs and platelet indices. Haematological analysis indicated substantial declines (p < 0.01) in total RBCs counts, PCV% and (Hb) in different degrees of anemic Theileriosis infected cattle compared with control group. In severe anemic group, MCV showed a substantial rise (p < 0.01) compared with moderate anemic group and (p < 0.05) compared with mild anemic and control groups; while MCH showed a substantial increase (p < 0.05) compared with mild and moderate anemic groups. In the coagulation profile, Platelets count (PLT) and PCT% showed insignificant difference (p > 0.05) among anemic groups compared with control group. MPV and PDW demonstrated substantial decreases (p < 0.01) in all anemic groups when compared with control group. In moderate anemic group, Prothrombin time (PT) exhibited a substantial prolongation (p < 0.01) compared to control group and (p < 0.05) compared to mild anemic group. In severe anemic group, PT was significantly prolonged (p < 0.05) compared with control and mild anemic groups. Correlations between RBCs and platelet indices in mild, moderate and severe anemic Theileriosis infected cattle revealed significant correlations in all blood parameters. It could be concluded that different degrees of anemia caused by T. annulata infection in cattle are associated with marked changes in measured haematological and coagulation parameters. [ABSTRACT FROM AUTHOR]

Background: Theileriosis is a haemoprotozoan illness that poses severe economic loss to dairy farmers due to its negative impact on mortality and productivity, especially in tropical and subtropical areas of the world. Methods: The present study was conducted for a period of one year (June 2020 to July 2021) during which 55 blood samples of cattle showing clinical signs suggestive of theileriosis were examined and quantified by using multiplex real time PCR for Theileria orientalis and Theileria annulata using Taq-Man probe. Result: Prevalence of theileriosis in three Theileria-endemic districts of Odisha due to Theileria annulata and Theileria orientalis were 26.11 per cent (11/42) and 57.14 per cent (24/42), respectively. Mixed infection were reported in 16.6 percent (7/42) cases. Parasite load for T. annulata in clinically affected cases ranged from 2.85X10³ to 1.51X105 parasites per ml of blood while for T. orientalis, it ranged from 1.94X104 to 5.88X105 parasites per ml of blood. Ten cattle positive for T. orientalis were administered two doses of buparvaquone (Butalex) @2.5 mg/kg b.w. at an interval of 48 hours after which degree of parasitemia were quantified ten days post treatment. The parasitic load in blood ten days post treatment decreased significantly from 4.04 = 104 to 2.21-102 per ml of blood. [ABSTRACT FROM AUTHOR]

Theileriosis, a tick borne protozoan disease, drastically affects the productivity of bovine with massive economic repercussion. The current investigation was conducted to determine the molecular prevalence of T. annulata employing cytochrome b gene based PCR assay. Investigation was conducted on 150 blood samples of cattle obtained from three districts, viz. Rajnandgaon (50), Balod (50) and Kabirdham (50) of Chhattisgarh state. Initially microscopic examinations of smears were done followed by PCR analysis of DNA samples. The overall prevalence of tropical theileriosis in plain region of Chhattisgarh was found to be 27.3% (41/150) and 54.0% (81/150) by microscopy and PCR assay, respectively. Among districts, highest prevalence 32% and 60% was recorded in Kabirdham followed by 22% and 54% at Balod and 28% and 50% at Rajnandgaon, by microscopy and PCR assay, respectively. Observations showed that, more number of infected animals were from unorganized farm (62.6%) than organized farm (45%). Present study revealed crossbred cattle more susceptible to T. annulata infection than indigenous animals. [ABSTRACT FROM AUTHOR]

For over a decade, bovine anaemia caused by Theileria orientalis Ikeda has been a significant disease in the Australian cattle industry. In this study, we conducted a spatial and temporal analysis of theileriosis in Australia using historic data from submissions to the New South Wales Department of Primary Industries (NSW DPI) from 2006 to 2022, where herd history, clinical signs, and PCR results were available. Since the first detections of bovine theileriosis in the Sydney area in 2006, the disease spread north- and southward and is now endemic to the southeast coast of Australia, closely mirroring the distribution of the principal vector Haemaphysalis longicornis. Across all years, the prevalence of the Ikeda genotype was 88%, while the prevalence of the benign Chitose and Buffeli genotypes was 55% and 38%, respectively. The majority of submissions were from beef cattle in coastal NSW, with anaemia, fever, jaundice, abortion, and lethargy the most frequently reported clinical signs. Transportation was identified as the major risk factor for disease. Until 2015, the majority of cases were reported in adult cattle, while in later years, calves made up the majority of cases, most likely due to the widespread acquisition of immunity in adults. Calves were significantly more likely to present with diarrhoea, lethargy, and anaemia, and to suffer mortality, while adults were significantly more likely to present with jaundice. Instances of abortion were observed to be significantly associated with beef cattle. The relationship between the level of parasitaemia and anaemia revealed a strong negative correlation for all animals examined. [ABSTRACT FROM AUTHOR]

Site‐selective C(sp3)−H functionalization of unreactive hydrocarbons is always challenging due to its inherited chemical inertness, slightly different reactivity of various C−H bonds, and intrinsically high bond dissociation energies. Here, a site‐selective C−H alkylation of naphthoquinone with unactivated hydrocarbons using Mn2(CO)10 as a catalyst under blue‐light (457 nm) irradiation without any external acid or base and pre‐functionalization is presented. The selective C−H functionalization of tertiary over secondary and secondary over primary C(sp3)−H bonds in abundant chemical feedstocks was achieved, and hydroxylation of quinones was realized in situ by employing the developed methodology. This protocol provides a new catalytic system for the direct construction of high‐value‐added compounds, namely, parvaquone (a commercially available drug used to treat theileriosis) and its derivatives under ambient reaction conditions. Moreover, this operationally simple protocol applies to various linear‐, branched‐, and cyclo‐alkanes with high degrees of site selectivity under blue‐light irradiated conditions and could provide rapid and straightforward access to versatile methodologies for upgrading feedstock chemicals. Mechanistic insight by radical trapping, radical scavenging, EPR, and other controlled experiments well corroborated with DFT studies suggest that the reaction proceeds by a radical pathway. [ABSTRACT FROM AUTHOR]

Antitheilerial resistance and treatment failure are growing international concerns. Against this backdrop, the first study aimed to compare the efficacy of Juglone (a naturally occurring naphthoquinone) and buparvaquone against bovine tropical theileriosis during natural infection. A total of 60 adult cattle (2-4 years) were selected and divided into five groups, comprising 15 animals each. The animals of Group-I were administered two doses of buparvaquone (BPQ) at a dose rate of 2.5mg/kg body weight intramuscular, 48 hours apart. The cattle of Group-II were administered with BPQ and oxytetracycline (OXY) at the dose rate of 2.5 mg/kg (two doses at 48-hour intervals) and 20 mg/kg intramuscular 3 doses at 48-hour intervals, respectively. While infected animals of Group-III were treated with Juglone (JUG) at a dose rate of 0.48 mg/kg IV on alternate days for 7 days. Whereas, Group-IV acted as a healthy control. The hemogram revealed normocytic normochromic anemia in infected animals. A significant decrease of neutrophils along with decrease of total leukocytes and lymphocytes was noticed in recovered animals from all treated groups (Group I to III). After 21 days, the results of the therapeutic trial showed the highest efficacy of Group II (BPQ+OXY) (93.3%; 14/15), followed by Group III (JUG) (73.3%) and Group I (BPQ) (66.7%; 10/15) groups, three weeks after treatment. It can be concluded that BPQ+OXY had the highest efficacy; whereas the efficacy of Juglone is also comparable, this can be used as an alternative treatment against tropical theileriosis. [ABSTRACT FROM AUTHOR]

Background: The production of bovine theileriosis vaccine involves in vitro cultivation of Theileria annulata schizont-infected cell lines. Fetal bovine serum (FBS) is commonly used in animal cell culture, including the Theileria cell line. However, we aimed to reduce the amount of serum needed for cell culture by modifying the Stoker culture medium with supplements such as excretion factor and serum substitutes. Methods: To evaluate the effectiveness of these modifications, techniques such as cell counting, cell viability assays, and genomic analysis were employed in the Parasitic Vaccines Production Department of Razi institute of Iran, from 2020 to 2022. Statistical analysis was used to compare the results of different experimental conditions. Results: The three experimental media were as effective as the commonly used 10% Stoker medium in supporting the growth and viability of cells. Conclusion: The significant reduction in the required amount of serum and the remarkable cell growth achieved by using defined serum replacements for the production of cell culture media is a significant step towards the preparation of a proper cell culture medium for the production of bovine Theileriosis vaccine. [ABSTRACT FROM AUTHOR]

The control of Tropical Theileriosis, a tick-borne disease with a strong impact on cattle breeding, can be facilitated using marker-assisted selection in breeding programs. Genome-wide association studies (GWAS) using high-density arrays are extremely important for the ongoing process of identifying genomic variants associated with resistance to Theileria annulata infection. In this work, single-nucleotide polymorphisms (SNPs) were analyzed in the Portuguese autochthonous cattle breeds Alentejana and Mertolenga. In total, 24 SNPs suggestive of significance (p ≤ 10−4) were identified for Alentejana cattle and 20 SNPs were identified for Mertolenga cattle. The genomic regions around these SNPs were further investigated for annotated genes and quantitative trait loci (QTLs) previously described by other authors. Regarding the Alentejana breed, the MAP3K1, CMTM7, SSFA2, and ATG13 genes are located near suggestive SNPs and appear as candidate genes for resistance to Tropical Theileriosis, considering its action in the immune response and resistance to other diseases. On the other hand, in the Mertolenga breed, the UOX gene is also a candidate gene due to its apparent link to the pathogenesis of the disease. These results may represent a first step toward the possibility of including genetic markers for resistance to Tropical Theileriosis in current breed selection programs. [ABSTRACT FROM AUTHOR]

Theileria annulata is an intracellular haemoprotozoan transmitted by ticks and its prevalence is reported mostly in the tropical region of the world. Tropical theileriosis mainly treated with Buparvaquone and tetracycline along with supportive therapy. Since there are reports on the development of resistance against buparvaquone so for that present study was carried out to evaluate the antitheilerial drug regime against the local strain of theileriosis. The present study on antitheilerial drug response was done to compare the combination therapy (Buparvaquone with tetracycline) and Buparvaquone only. A total of 18 Buffalo were selected from Ayodhya a Eastern Plane Zone (EPZ) of Uttar Pradesh and separated into three groups each with 6 animals. Treatment Group I and Group II were comprised of infected animals and group III included the healthy animals only. Before the experiment, at zero-day all 18 animals were screened for Theileria annulata infection by microscopic examination, and confirmed by Tams1 gene-based Polymerase Chain Reaction (PCR). Further, the same procedure followed for the blood for the samples collected on different days of post-treatment. Group I animal received the dose of buparvaquone @2.5mg/kg.b.wt. i.m., whereas the group II animals received the dose of buparvaquone @2.5mg/kg.b.wt. i.m., with oxytetracycline @20mg/kg.b.wt. i.v. twice within 48 hours. Hematological (Hb, PCV, TEC, DLC) and biochemical variables (total serum protein, albumin, blood urea nitrogen, AST, ALT) were assessed before and after treatment. After treatment the level of haemato-biochemical parameters restored early in the group II treated with combination therapy of Buparvaquone with tetracycline than in group I treated with Buparvaquone only. Moreover, the results of the treatment and control groups were significantly (<0.05) different with each other. The present study revealed that the treatment of Theileria annulata infected Buffalos with the combination of buparvaquone and oxytetracycline provides slightly better recovery than the treatment with buparvaquone only (group I). However, treatment with Buparvaquone or combination with tetracycline repeated after 48 hours both worked well against tropical theileriosis prevalence. [ABSTRACT FROM AUTHOR]

ABSTRACT Theileria annulata is a tick-transmitted apicomplexan parasite that gained the unique ability among parasitic eukaryotes to transform its host cell, inducing a fatal cancer-like disease in cattle. Understanding the mechanistic interplay between the host cell and malignant Theileria species that drives this transformation requires the identification of responsible parasite effector proteins. In this study, we used TurboID-based proximity labeling, which unbiasedly identified secreted parasite proteins within host cell compartments. By fusing TurboID to nuclear export or localization signals, we biotinylated proteins in the vicinity of the ligase enzyme in the nucleus or cytoplasm of infected macrophages, followed by mass spectrometry analysis. Our approach revealed with high confidence nine nuclear and four cytosolic candidate parasite proteins within the host cell compartments, eight of which had no orthologs in non-transforming T. orientalis. Strikingly, all eight of these proteins are predicted to be highly intrinsically disordered proteins. We discovered a novel tandem arrayed protein family, nuclear intrinsically disordered proteins (NIDP) 1–4, featuring diverse functions predicted by conserved protein domains. Particularly, NIDP2 exhibited a biphasic host cell-cycle-dependent localization, interacting with the EB1/CD2AP/CLASP1 parasite membrane complex at the schizont surface and the tumor suppressor stromal antigen 2 (STAG2), a cohesion complex subunit, in the host nucleus. In addition to STAG2, numerous NIDP2-associated host nuclear proteins implicated in various cancers were identified, shedding light on the potential role of the T. annulata exported protein family NIDP in host cell transformation and cancer-related pathways.IMPORTANCETurboID proximity labeling was used to identify secreted proteins of Theileria annulata, an apicomplexan parasite responsible for a fatal, proliferative disorder in cattle that represents a significant socio-economic burden in North Africa, central Asia, and India. Our investigation has provided important insights into the unique host-parasite interaction, revealing secreted parasite proteins characterized by intrinsically disordered protein structures. Remarkably, these proteins are conspicuously absent in non-transforming Theileria species, strongly suggesting their central role in the transformative processes within host cells. Our study identified a novel tandem arrayed protein family, with nuclear intrinsically disordered protein 2 emerging as a central player interacting with established tumor genes. Significantly, this work represents the first unbiased screening for exported proteins in Theileria and contributes essential insights into the molecular intricacies behind the malignant transformation of immune cells.

Immunisation of livestock with high-quality vaccines and the use of acaricides are highly ranked tick control strategies worldwide. However, the effects of coupling livestock immunisation and acaricide control on mitigating the spread of East Coast Fever (ECF) is not well understood. Effective strategies to curb the disease require an understanding of the influence of control strategies on ECF dynamics. This paper presents a new mathematical model for ECF in ticks and livestock to analyze the effect of livestock immunisation and acaricide control on preventing ECF spread. Our research is focused on examining how vaccine efficacy, inoculation rate, and acaricide efficacy affect disease progression. Our finding is that acaricide control alone may not be sufficient to stop the spread of ECF, even if it has an 80% effectiveness all the time. However, by pairing acaricide control with livestock vaccination, disease transmission is significantly reduced and elimination is possible under certain circumstances. Overall, results show that it is crucial to understand the influence of combining control strategies to mitigate the spread of this devastating livestock disease and enhance decision making among policymakers and livestock keepers.

Theileriosis is one of the crucial vector-transmitted diseases in cross-bred dairy cattle in India and it is commonly caused by Theileria annulata. Frequently reported clinical signs in theileriosis were pyrexia, lymphadenopathy, conjunctival petechiae, and anemia. The purpose of the present communication report the anterior uveitis and bilateral corneal opacity in young crossbred calves due to theileriosis. Two Jersey crossbred calves aged between 22 to 40 days were presented to the clinic with a history of inappetence, pyrexia, severe tick infestation, corneal opacity, and proptosis. Ultrasonography of the eye revealed anterior uveitis and peripheral blood smears revealed the presence of piroplasms of Theileria in the erythrocytes and further confirmation was done by PCR assay. The calves were treated with injections of buparvaquone, flunixin meglumine, and vitamin A along with eye drops containing gatifloxacin, flurbiprofen, and carboxy methyl cellulose sodium.

Background. Cattle anaplasmosis and babesiosis and theileriosis of horses are a group of natural focal vector-borne diseases that cause significant economic damage to livestock and horse breeding around the world. On the territory of the Baikal region these diseases and their pathogens remain little explored. Purpose – to study the epizootic situation of blood-parasitic diseases of farm animals in the Irkutsk region, to establish the species affiliation of the identified pathogens and their genetic diversity. Materials and methods. For the presence of Babesia spp./Theileria spp. and Anaplasma spp. has been researched659 horse blood samples, 579 sheep blood samples, 25 goat blood samples and 647 cattle blood samples. For the detection of babesia and theileria, nested two-round PCR with primers from the 18S rRNA gene region was used, and for the detection of anaplasma DNA with primers from the 16S rRNA gene region. For study the genetic diversity of the identified pathogens, selective sequencing of samples was carried out with subsequent phylogenetic analysis. Results. The DNA of Anaplasma spp. was detected in blood samples of small ruminants from 12 districts of the region. The infection rate of sheep and goats was 68.7±1.9% and 68%±9.3%, respectively. Nucleotide sequences of the 16S rRNA gene fragment were determined in 68 A. ovis samples from the blood of sheep and goats. The researched sequences were conservative for the studied gene and differed from each other in two heterozygous sites of the gene(T/C and G/A nucleotide substitutions).They corresponded to the sequences of the prototype strain Haibei (CP015994), as well as to the A. ovis sequences previously found in the blood of sheep from Altai (Russia) and Mongolia, deer and D. niveus and D. nuttalli ticks from China. DNA of Babesia spp./Theileria spp. found in blood samples of horses from 13 districts of the region. The average infection of horses was 64.2%±1.9%. Two etiological agents of equine piroplasmosis were found in the blood of the examined horses: T. equi (genetic groups A and E) and B. caballi. Definite sequences of the B. caballi 18S rRNA gene (700–1146 bp) were identical to each other and differed by at least ten substitutions and one deletion from the sequences available in the GenBank database. However, the greatest similarity was observed with B. caballi sequences found in the blood of a horse from Brazil (KY952238), ticks from Kazakhstan (MN907451) and China (MN173021). The results indicate that DNA of Anaplasma and Babesia in the blood of cattle have not been identified. Conclusion: The data received evidence a wide distribution of foci of small cattle anaplasmosis and piroplasmosis of horses in the Irkutsk region and the need to continue monitoring their epizootic condition.

In this study, examination of infected cattle infested with tick, identification of collected tick samples were based on the12S rDNA PCR products as Rhipicephalus annulatus, the GenBank accession number is (OP650242). A total of 72 blood samples from crossbred cattle of both sexes were examined clinically and in the laboratory. Out of these, 43 cattle were healthy, while 19 (26.38%) had theileriosis and 10 (13.88%) had babesiosis. Hemogram analysis revealed distinct anemia patterns, with Babesia-infected cattle displaying macrocytic hypochromic anemia and Theileria-infected ones showing normocytic normochromic anemia, both with reduced platelet counts. Babesia-infected cattle had elevated total leukocyte counts, neutrophilia, eosinophilia, and lymphopenia, while Theileria-infected cattle had decreased total leukocyte counts, neutropenia, lymphocytosis, and eosinophilia. In infected cattle, serum biochemistry showed increased ALT, AST, creatinine, and urea levels in both Babesia and Theileria infections. There was decreased serum protein, and albumin, in both cases. Oxidative stress revealed elevated serum malonaldehyde (MDA), reduced glutathione peroxidase (GPx) and catalase (CAT) levels in infected animals compared to controls. After administering Imidocarb dipropionate (1mg/kg S/C) and Buparvaquone (1ml/20kg I/M) to animals with babesiosis and theileriosis, respectively, there was a positive change in the hematological and biochemical measures, bringing them closer to the normal values. There is a genuine danger to the cattle industry in Egypt due to the existence of babesiosis, theileriosis, and their vector. Modern techniques like PCR should be utilized for precise monitoring and to prevent spread of such diseases. Furthermore, adverse effect of Babesia and Theileria on hematological and biochemical parameters can be eliminated through the appropriate use of Imidocarb dipropionate and Buparvaquone for babesiosis and theileriosis respectively. [ABSTRACT FROM AUTHOR]

The present case reports an atypical form of theileriosis which is seldom seen in field condition. A 9-year-old cattle brought to veterinary hospital, teaching veterinary clinical complex of University with a history of multiple subcutaneous nodules (2–4 cm) on the upper half, fever and decreased milk production for the past 10 days. Detailed clinical examination revealed pale mucous membranes, enlargement of superficial lymph nodes and pyrexia. Fine needle aspiration cytology of swollen lymph nodes revealed an increased number of lymphoid cells. Haematology revealed mild anaemia, leucopenia and typical piroplasm in peripheral blood smear. DNA was extracted from the suspected blood sample which was subjected to PCR assay for diagnosis of theileriosis and lumpy skin disease. Blood PCR assay results showed sample positive for Theileria annulata and negative for lumpy skin disease virus. Specific treatment with buparvaquone at 2.5 mg/kg was given along with supportive treatment. Spontaneous clinical recovery and improvement in haematocrit was observed day 5 post therapy. In conclusion, in case of dermatological nodular skin lesion in cattle theileriosis should be considered as one of the tentative aetiology and differential diagnosis should be done with lumpy skin disease and lymphosarcoma. [ABSTRACT FROM AUTHOR]

Theileriosis is one of the crucial vector-transmitted diseases in cross-bred dairy cattle in India and it is commonly caused by Theileria annulata. Frequently reported clinical signs in theileriosis were pyrexia, lymphadenopathy, conjunctival petechiae, and anemia. The purpose of the present communication report the anterior uveitis and bilateral corneal opacity in young crossbred calves due to theileriosis. Two Jersey crossbred calves aged between 22 to 40 days were presented to the clinic with a history of inappetence, pyrexia, severe tick infestation, corneal opacity, and proptosis. Ultrasonography of the eye revealed anterior uveitis and peripheral blood smears revealed the presence of piroplasms of Theileria in the erythrocytes and further confirmation was done by PCR assay. The calves were treated with injections of buparvaquone, flunixin meglumine, and vitamin A along with eye drops containing gatifloxacin, flurbiprofen, and carboxy methyl cellulose sodium. [ABSTRACT FROM AUTHOR]

Tropical or Mediterranean theileriosis in dairy cattle is widely distributed in many tropical regions of the world. The purpose of this study was to evaluate the proliferation status of mononuclear cells infected with Theileria annulata schizonts in different tissues and its relationship with the pathogenesis of the parasite in cattle by histopathology, immuno-histochemistry and polymerase chain reaction (PCR). Blood and tissue samples of eight Holstein cattle that had been lost due to theileriosis and eight healthy slaughtered cattle of the same breed were collected as a control group after necropsy. The piroplasms in the blood smears and the schizonts in the cytoplasm of the lymphocytes and macrophages of the lymph nodes were microscopically detected. Histopathologically, the proliferation of macrophages, lymphocytes, and plasma cells in lymph nodes and the heart, congestion, and bleeding in the red pulp of the spleen, portal tracts of the liver, interstitial tissue of the kidneys, multifocal necrosis and ulceration in the abomasum together with hyperemia and hemorrhages and lymphoblastic infiltration in the submucosa and lamina propria adjacent to these lesions and emphysema with ecchymotic hemorrhage in the lungs were evident. Immunohistochemistry identified the proliferated cells as mostly Cluster of Differentiation 3- Positive T lymphocytes and macrophage marker antibody 387- positive macrophages. Positive results of PCR for the Tamsl 30.00 kDa gene were observed in lymph nodes, liver, lung and abomasum. It was concluded that the pathological changes were the result of schizont-infected macrophage proliferation leading to severe uncontrolled proliferation of uninfected T lymphocytes. [ABSTRACT FROM AUTHOR]

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Background: Ovine and caprine theileriosis is a tick-borne hemoprotozoan disease, caused by Theileria spp., responsible for heavy economic losses in terms of high mortality and morbidity rates. Diagnosis of ovine theileriosis is primarily based on clinical symptoms, microscopic screening of stained blood smears, and lymph node biopsy smears, but the limitations of these detection methods against Theileria spp. infection limits their specificity. Aim: To overcome these limitations, the current study reports the differential diagnosis of theileriosis through a blood smear examination and polymerase chain reaction (PCR) in small ruminants from Pakistan. Methods: The study was conducted on 1,200 apparently healthy small ruminants (737 sheep and 463 goats). First, blood smears were screened for the presence of Theileria piroplasms in red blood cells. Second, PCR amplification based on 18S rRNA gene was performed by using primers specific to Theileria spp. Results: Out of the 1,200 samples of examined blood smears, 100 animals (8.33%) were found positive for Theileria species, which showed intra-erythrocytic bodies in the form of dot and comma shapes. Amplification of the isolated DNA from randomly collected blood samples of 737 sheep and 463 goats showed that an amplicon size of 1,098 bp was positive for Theileria spp. In total, 315 out of the 1,200 small ruminants examined in this study were found positive for Theileria spp. DNA through PCR amplification. Notably, out of the 885 blood samples negative by PCR amplification, only 15 blood samples were found positive by the blood smear test. Conversely, 230 blood samples that tested negative in the smear technique produced a specific band through PCR amplification. Overall, the sensitivity and specificity rates were 26.98% and 98.31% for the blood smear method and 73.01% and 100% for the PCR assay, respectively. Conclusion: Our finding suggests that PCR is the gold standard method compared to the conventional method of smear examination for the diagnosis of ovine and caprine theileriosis in Pakistan. [ABSTRACT FROM AUTHOR]

Background: Tropical theileriosis, Theileria annulata infection, is the most prevalent summer disease in Tunisia. It is transmitted by Hyalomma scupense, a two‐host tick known to be endophilic. Objectives: The present study aimed to estimate the infection prevalence of cattle by T. annulata in two districts from central Tunisia. Methods: Blood samples collected from 270 Holstein cattle from the Sidi Bouzid (140 samples) and Kasserine districts (130 samples) were analysed by Giemsa staining and T. annulata‐specific PCR. Results: In both regions, PCR revealed a prevalence of 32.6%. This was significantly higher than the 6.3% prevalence obtained by Giemsa staining blood smears (p < 0.001). Giemsa staining also revealed a low parasitaemia of 0.05%. The PCR‐based prevalence was not statistically different between the two districts (31.4 ± 0.04 and 33.8 ± 0.04% in Sidi Bouzid and Kasserine districts, respectively, p = 0.6). On the contrary, the results of blood smear examination (2.85 and 10% in Sidi Bouzid and Kasserine, respectively) differed significantly between the two sampling sites (p = 0.01). There was no evidence of a statistically significant difference between the overall molecular infection prevalence when the samples were segregated based on animals' age or gender (p = 0.1 and 0.2, respectively) and a similar trend was observed for Giemsa staining. Ten PCR amplicons of the Tams1 gene (721 bp) were subsequently sequenced from the two regions. The phylogenetic analyses showed 100% similarity between all sequences. The unique conserved Tams1 sequence was deposited in GenBank under the accession number OP428816 and used to infer its phylogenetic relationships with those available in the GenBank repository. Conclusions: This is the first report of the presence of T. annulata in this region of central Tunisia which has no history of tropical theileriosis. Priority areas for future studies include understanding the origin of these T. annulata‐positive animals in a region where the presence of a known natural vector tick, H. scupense, has not been reported. Given that the disease severely constrains cattle productivity, it would also be worthwhile to investigate if other potential vectors for T. annulata, such as Hyalomma dromedarii, are present in the arid regions. [ABSTRACT FROM AUTHOR]

Theileria sp. (Piroplasmida: Theileriidae) is one of the most widely known infections transmitted by hard ticks (Acari: Ixodidae) and has been linked to significant economic losses across the globe. The study's main emphasis was theileriosis, a disease that is common in Pakistan and has an incidence ranging from 0.6% to 33%. Through DNA screening of the vector ticks and host blood, this study sought to determine the risk of tick-borne theileriosis in populations of buffalos (Bubalus bubalis) and cattle (Bos indicus) in Toba Tek Singh district of Punjab, Pakistan. Identified tick species include Hyalomma anatolicum (35.4%), Rhipicephalus (Boophilus) microplus (30.2%), and R. sanguineus (25%). Tick specimens were collected from animals and their respective microenvironments. PCR assays targeting Theileria annulata were used to investigate the infection in the DNA extracted from the collected blood samples from large ruminants and salivary glands (SGs) of the Hyalomma ticks. The 18S rRNA of T. annulata was amplified using specific primers. Positive T. annulata amplicons were sequenced and verified using BLAST analysis. Overall, 50% of SGs contained T. annulate DNA. Female ticks, and those collected from cattle and from riverine environments had significantly higher (p < 0.05) rates of Theileria infection in their acini. Overall prevalence of Theileria infection was 35.9% in blood collected from large ruminants. Cattle had a substantially greater frequency of bovine theileriosis (43.2%) than buffalos (28.7%). Age and sex of large ruminants were significantly positively associated (p < 0.05) with Theileria infection. Furthermore, compared to non-riverine cattle (35%) and buffalo (19.5%), riverine cattle (52.2%) and buffalo (36.2%) showed a considerably higher prevalence. The results of this study, which is the first in Pakistan to examine the blood of large ruminants and vectorial function of Ixodid ticks in the transmission of T. annulata along with associated risk factors, offer an important insight for risk assessment of Theileria infection in livestock using vectorial infectivity. [ABSTRACT FROM AUTHOR]

The current work was conducted to unveil the current situation for the infection by Theileria annulata in cattle in Al-Hilla City, Iraq. A total of 225 blood samples (200 from suspected infected animals and 25 from clinically healthy animals as a control group) were collected. These samples were subjected to a direct slide-smearing for detection using a microscope and DNA sequencing, targeting the cytochrome b (Cyt b) gene of 10 polymerase chain reaction (PCR) products. The thin smear findings of the 200 suspected cases revealed that 63 (31.5%) were infected with Theileria spp., while 115 (57.5%) cases had no Theileria but other blood parasites; however, only 22 (11%) suspected cases showed no presence of any parasites. Unsurprisingly, the 25 blood samples from the control group demonstrated no presence of any blood parasite. Moreover, the DNA sequencing demonstrated that the Theileria spp. belonged to T. annulata species, and these sequences were nucleotide-based similar to Gene-Bank isolates from Tunisia (ON035604, ON035605, ON035606, ON035607, ON035608, ON035609, ON035610, ON035611, ON035612, and ON035613). The present study outcomes indicate that theileriosis is the dominant parasitic infection in cattle in Al-Hilla City and is highly caused by Theileria annulata.

It is established thitithet takes place the wide circulation of horned cattle teileriosis in the areas of Samarkand, Navoi, Djisak regions. Which differ with its environmental conditions? At the same taim there is distingulcheda a low virulentb strain from the jf Samarkand region, which is sultable for manufacturing vacsinis against theileriosis. [ABSTRACT FROM AUTHOR]

Wrong identification and characterization of Theileria species in cattle may be associated with economic loss via low productive rate in cattle business. A study was designed to compare microscopy and polymerase chain reaction (PCR) in identification and characterization of Theileria species of cattle within Makurdi, Benue State Nigeria. Blood samples were collected from cattle of both sexes and analysed microscopically using thin blood film and DNA examination was done using polymerase chain reaction (PCR). Theileria species were identified and characterized by PCR amplification using species specific primers of 18s rRNA for Theileria Spp. Data obtained were analysed using chi square, and P values at less than 0.05 were considered significant different. The result revealed the presence of Theileria. The result of comparison of prevalence of infection of Theileriosis using microscopy and PCR revealed that, microscopy was able to detect only 1.77 % of Theileria but PCR detected 13.3% of Theileria. The percentage of prevalence of Theileria species was highly detected by PCR than microscopy and this was statistically significant (P < 0.05). Theileriosis in cattle have been seen to be prevalence in Makurdi metropolis and molecular method such as PCR can effectively. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate an outbreak of Theileria annulata (T. annulata) infection in an organized dairy cattle farm in Madhya Pradesh, India, using clinical and molecular techniques. Following the deaths of two crossbred cattle in March 2021, 43 blood samples were collected from infected and apparently healthy animals and examined by blood smear and polymerase chain reaction (PCR) techniques. The blood smear examination showed that 23.25% of samples were positive for Theileria organisms, while conventional PCR targeting the 18S ribosomal RNA (18S rRNA) and T. annulata merozoite surface antigen-1 (TAMS-1) genes revealed that 32.55% of samples were positive for T. annulata. PCR targeting cytochrome b (Cytb) gene showed 46.51% of samples were positive for T. annulata. Haematological analysis confirmed clinical signs of infection in affected animals, which were treated with buparvaquone @ 2.5 mg/kg body weight intramuscularly along with supportive medicine. Two 18S rRNA gene amplicons were sequenced and analysed in a phylogenetic tree and haplotype network with 54 Indian and 38 foreign sequences. The phylogenetic tree revealed two groups with a high posterior probability and bootstrap value, while the haplotype network revealed 35 haplotypes, with haplotype 1 (H1) being the most abundant and several single haplotypes clustering around it, indicating fast and widespread expansion. Genetic diversity indices and neutrality tests confirmed that the population was expanding. These studies highlight the significance of prompt and precise diagnosis and management of T. annulata outbreaks and provide insights into its evolutionary history and population dynamics of T. annulata in India, which could aid improving disease preventive and control strategies. [ABSTRACT FROM AUTHOR]

The molecular changes occurring in the host in response to in vivo Theileria annulata parasitic infection are not well understood. Therefore, the present study investigated the differential expression profiles of peripheral blood mononuclear cells (PBMCs) across Theileria annulata-infected and non-infected crossbred cows. The differential expression profiles from PBMCs of infected and non-infected crossbred cows were generated by RNA sequencing. A marked difference in the expression of genes associated with innate immunity (FTH1, ACTB, ISG15) was observed between the two groups. The over-represented pathways in Theileria annulata-infected cows were associated with the immune system and regulation of the mitotic cycle. Enriched genes and pathways in non-infected animals were associated with the maintenance of chromatin integrity and cell structure. The highly connected genes identified in this study form potential candidates for further investigation into host–parasite interactions in cattle. An improved understanding of the transcriptomic dynamics during theileriosis would lead to underpinning molecular level differences related to the health status of cattle. [ABSTRACT FROM AUTHOR]

Context: Malignant ovine theileriosis is a tick-borne disease of sheep and goats, caused by protozoan Theileria lestoquardi. The disease has serious economic implications for small ruminant production around the world. Methods: An outbreak of malignant ovine theileriosis in a sheep flock was investigated from Hisar district of Haryana, India, in March 2022. The etiological agent was identified using polymerase chain reaction assay with genus specific primers targeting 18S rRNA gene and subsequently confirmed by sequencing. Results: The morbidity, mortality and case fatality rate reported in the outbreak were 22.2, 18.8 and 85%, respectively. The phylogenetic analysis clustered the present study T. lestoquardi isolate in the same clade with T. lestoquardi from Iraq, Iran and Pakistan with maximum nucleotide identity of 99.37% with strains from Iraq. The tick vector Hyalomma anatolicum recovered from dead animals was implicated in the disease's transmission. Conclusions: Malignant ovine theileriosis resulted in high case fatality rate. This study presents the first molecularly confirmed outbreak of malignant ovine theileriosis outbreak in the North Indian region, with characteristic post-mortem findings. [ABSTRACT FROM AUTHOR]

Tropical theileriosis is a protozoan infection caused by Theileria annulata, which significantly affects cattle worldwide. This study was aimed to analyze the TaSPAG1 protein and design a novel multi-epitope vaccine candidate. Online tools were employed for the prediction of Physico-chemical properties, antigenicity, allergenicity, solubility, transmembrane domains and signal peptide, posttranslational modification (PTM) sites, secondary and tertiary structures as well as intrinsically disordered regions, followed by identification and screening of potential linear and conformational B-cell epitopes and those peptides having affinity to bind bovine major histocompatibility complex class I (MHC-I) molecules. Next, a multi-epitope vaccine construct was designed and analyzed. This 907-residue protein was hydrophilic (GRAVY: -0.399) and acidic (pI: 5.04) in nature, with high thermotolerance (aliphatic: 71.27). Also, 5 linear and 12 conformational B-cell epitopes along with 8 CTL epitopes were predicted for TaSPAG1. The 355-residue vaccine candidate had a MW of about 35 kDa and it was antigenic, non-allergenic, soluble and stable, which was successfully interacted with cattle MHC-I molecule and finally cloned into the pET28a(+) vector. Further wet studies are required to assess the vaccine efficacy in cattle. [ABSTRACT FROM AUTHOR]

The study was carried out to study the prevalence of piroplasmid infections in the cattle population of Mathura. A total of 143 blood samples were randomly collected from apparently healthy animals under small-scale farming in four villages. A molecular prevalence study was conducted using the 18S rRNA gene based PCR reactions. The overall prevalence of piroplsmids was recorded at 27.97%. Results of the prevalence study revealed a high risk of infection in crossbred and young age animals during the monsoon season. In the studied animals, the piroplasmid stages of Theileria annulata and Babesia bigemina were present in 23.07% and 4.89% of samples, respectively. Molecular detection of piroplasmids was also conducted in ticks infesting cattle. The results of the study corroborate the role of R. microplus and H. anatolicum ticks in the transmission of babesiosis and theileriosis, respectively. Two positive samples for B. bigemina were sequenced for a partial sequence of 18S rRNA gene and subjected to phylogenetic analysis. B. bigemina of Mathura isolate had similarities with Sri Lanka and Vietnam isolate. This study reports the genetic diversity of B. bigemina and thus will help understand the molecular evolutionary history and the emergence and spread of new strains in the area under study. The results of this study will be helpful in further molecular surveillance studies and thereby in the effective control of these diseases. [ABSTRACT FROM AUTHOR]

Theileriosis is a tick‐borne protozoal disease caused by a piroplasm of the genus Theileria. Hard ticks are obligate hematophagous ectoparasites that serve as vectors of Theileria spp. Studies of the presence of theileriosis in Egyptian dogs and associated ticks are scarce. This study was conducted to detect and identify Theileria spp. in dogs and Rhipicephalus sanguineus ticks and to monitor the epidemiological data of this disease. The prevalence rates of Theileria equi infection were 12.02%, 0.73%, 2.93%, and 1.83% by microscopic examination of dog blood, tick hemolymph, tick midgut, and tick salivary smears, respectively. Conversely, the T. equi prevalence in dog blood and associated ticks assessed by PCR was 25.81% and 10.42%, respectively. Epidemiological data about Theileria infection revealed a significant difference in the infection between different seasons and different dog breeds (p value <0.05), whereas host, sex, and age of dogs had no significant effect on the infection. Sequencing of PCR products showed that all PCR positive samples were infected with T. equi. Transmission electron microscopy (TEM) described the different stages of Theileria in the midgut and salivary gland of infected ticks. The current study confirmed that T. equi is not specific to equine hosts, and confirmed that dogs are a susceptible host to T. equi. [ABSTRACT FROM AUTHOR]

Climate directly influences the epidemiology of vector-borne diseases at various spatial and temporal scales. Following the recent increased incidences of theileriosis in Zimbabwe, a disease mainly transmitted by Rhipicephalus appendiculatus, we determined lethal temperatures for the species and current and possible future distribution using the machine learning algorithm 'Maxent'. Rhipicephalus appendiculatus larvae had an upper lethal temperature (ULT50) of about 44 ± 0.5 °C and this was marginally higher for nymphs and adults at 46 ± 0.5 °C. Environmental temperatures recorded in selected zonal tick microhabitats were below the determined lethal limits, indicating the ability of the tick to survive these regions. The resultant model under current climatic conditions showed areas with high suitability indices to the eastern, northeastern and southeastern parts of the country, mainly in Masvingo, Manicaland and Mashonaland Central provinces. Future predictions as determined by 2050 climatic conditions indicate a reduction in suitable habitats with the tick receding to presently cooler high elevation areas such as the eastern Highlands of Zimbabwe and a few isolated pockets in the interior of the country. Lowveld areas show low suitability under current climatic conditions and are expected to remain unsuitable in future. Overall, the study shows that R. appendiculatus distribution is constrained by climatic factors and helps identify areas of where occurrence of the species and the disease it transmits is highly likely. This will assist in optimizing disease surveillance and vector management strategies targeted at the species. [ABSTRACT FROM AUTHOR]

Background and Aim: Theileriosis, caused by tick-borne hemoprotozoans of the genus Theileria, severely impacts the economics of the livestock industry in most tropical and subtropical countries. The aim of the present study was to detect Theileria spp. in domesticated animals (camels, cows, sheep, and goats) using direct microscopy and to determine the infection rate in geographically distinct regions in the northern emirates of the UAE. Materials and Methods: Blood samples (n = 536) were collected from clinically symptomatic and asymptomatic domesticated animals and subjected to Giemsa staining and examined microscopically for the identification of Theileria. Results: Smears showed an overall rate of positivity for Theileria spp. in 325/536 (60.6%) animals. Different infection rates were recorded across the various animal groups in the different study areas (Middle region 215/386 [55.7%], East region 100/139 [71.9%]). Of the 11 goat samples collected from the North region, 10 (90%) were positive. Infection rates per animal group based on microscopy were as follows: camels, 3/35 (8.5%); cows, 19/36 (52.7%); goats, 200/303 (66%); and sheep, 103/162 (63.5%). Real-time polymerase chain reaction confirmation of all microscopy-positive samples identified 23/325 (7.1%) results as false-positive. Conclusion: This study clarified that Theileria spp. is present in the Middle (Sharjah, Umm Al Quwain, and Ajman), East, and North regions. This report also confirmed the use of direct microscopy with Giemsa-stained blood films as the method of choice for diagnosing acute infections. Further work is needed to molecularly determine the prevalence and species of Theileria spp. circulating in the different parts of the UAE. [ABSTRACT FROM AUTHOR]

Theileriosis is a tick-borne disease that causes enormous losses in the dairy industry. There are several species of Theileria that can infect bovines. Generally, more than one species are prevalent in any geographical area; thus, chances of co-infections are high. Differentiation of these species may not be possible by microscopic examination or serological tests. Therefore, in this study, a multiplex PCR assay was standardized and evaluated for rapid and simultaneous differential detection of two species of Theileria viz., Theileria annulata and Theileria orientalis. Species-specific primers were designed to target the merozoite piroplasm surface antigen gene (TAMS1) of T. annulata and the major piroplasm surface protein gene of T. orientalis, yielding specific amplicon of 229 bp and 466 bp, respectively. The sensitivity of multiplex PCR was 102 and 103 copies for T. annulata and T. orientalis, respectively. The simplex and multiplex PCRs were specific and showed no cross-reactivity with other hemoprotozoa for either primer. For comparative evaluation, blood samples from 216 cattle were tested by simplex and multiplex PCR for both species. Using multiplex PCR, 131 animals were found infected for theileriosis, of which 112 were infected with T. annulata, five were infected with T. orientalis, and 14 had mixed infections. This is the first report of T. orientalis from Haryana, India. Representative sequences of T. annulata (ON248941) and T. orientalis (ON248942) were submitted in GenBank. The standardized multiplex PCR assay used in this study was specific, sensitive, for the screening of field samples. [ABSTRACT FROM AUTHOR]

Schizont-infected nucleated cells and anemia-induced hypoxia may cause acute kidney injury (AKI) in cattle with tropical theileriosis. In most cases, serum creatinine (sCr) fails to detect early kidney injury. Recently, urine biomarkers such as neutrophil gelatinase-associated lipocalin (NGAL), Cystatin C (CysC), and Clusterin (Clu) have outperformed sCr in identifying AKI in various animal species. However, these markers have not been tested in cattle for AKI detection. This study investigated AKI in cattle with tropical theileriosis using serum and urinary NGAL, CysC, and Clu and compared them to standard renal function markers such as sCr, urine protein to creatinine (uPC) ratio and urine-specific gravity (USG). Seventeen cattle were infected with Theileria annulata (Th) and ten were controls (CTRL) in this investigation. The urinary NGAL (uNGAL) in the Th group was considerably (P<0.001) greater than that in the CTRL group. uCysC increased significantly in the Th group compared to CTRL group (P<0.001). The Th group had higher sClu than the CTRL group (P<0.001). There was a significant difference (p=0.005) in sCr concentrations between Th and CTRL groups, but the values were within the reference range in both groups. Th group had a higher uPC ratio than CTRL (p= 0.002), whereas USG was lower in Th group than in CTRL group (P<0.001). These results suggested that cattle with tropical Theileriosis have acute tubular injury. [ABSTRACT FROM AUTHOR]

Theileriosis is one of the most important tick-borne diseases that has been affecting farmers and thousands of livestock in Zimbabwe. The main government strategy to combat theileriosis is use of plunge dips with anti-tick chemicals at specified times; however, an increase in number of farmers caused a strain on government services resulting in disease outbreak. One of the key issues that have been highlighted by department of veterinary is the strain in communication and knowledge of the disease with the farmers. Hence, it is important to evaluate the communication between farmers and veterinary services and identify possible areas of strain. A field survey was conducted with 320 farmers in Mhondoro Ngezi, a district badly affected by theileriosis. Face-to-face interviews with smallholders and communal farmers were conducted between September and October 2021, and the data were analyzed using Stata 17. Communal farmers relied mainly on oral communication and had limited knowledge of theileriosis; therefore, dead cattle % was high among them. Though veterinary extension officers were the prime source of information, oral communication medium affected knowledge transferred. The results of this study recommend adoption of communication mediums that encourage retention, such as brochures and posters by veterinary extension services. The government may also partner with private players to ease pressure of increased farming population due to land reform. [ABSTRACT FROM AUTHOR]

Accurate quantification based on nucleic acid amplification is necessary to avoid the spread of pathogens, making early diagnosis essential. Droplet digital PCR (ddPCR) stands out for absolute parasite quantification because it combines microfluidics with the TaqMan test. This helps deliver maximum accuracy without needing a reference curve. This study assessed the efficacy of ddPCR as a detection tool for the bovine theileriosis (BT) caused by Theileria parasites. We developed and validated a duplex ddPCR method that detects and quantifies the Theileria genus (18S rRNA) and identifies clinically significant Theileria annulata parasites (TaSP) in experimental and clinical samples. ddPCR was shown to be as effective as qPCR throughout a 10-fold sample dilution range. However, ddPCR was more sensitive than qPCR at lower parasite DNA concentrations and reliably assessed up to 8.5 copies/µL of the TaSP gene in the infected DNA (0.01 ng) samples. The ddPCR was very accurate and reproducible, and it could follow therapeutic success in clinical cases of theileriosis. In conclusion, our ddPCR assays were highly sensitive and precise, providing a valuable resource for the study of absolute parasite quantification, drug treatment monitoring, epidemiological research, large-scale screening, and the identification of asymptomatic parasite reservoirs in the pursuit of BT eradication. [ABSTRACT FROM AUTHOR]