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This study is the first to conduct a sero-surveillance of Bovine Tropical Theileriosis (BTT) caused by the protozoan parasite Theileria annulata (T. annulata) using a recombinant Tams1 protein-based dot-ELISA in cattle, and to compare its efficacy with plate-ELISA, single PCR, nested PCR, and blood microscopy. The goal was to identify the most effective method for the early and accurate detection of theileriosis, which significantly impacts livestock through reduced milk yield and increased mortality. A total of 101 field blood samples were examined using blood smear analysis, single PCR, nested PCR, and dot-ELISA. The recombinant Tams1 protein was successfully cloned and expressed using a pET-30b (+) expression vector in a prokaryotic system. The protein was purified with Ni-NTA chromatography, confirmed for immunoreactivity with T. annulata positive serum via Western blot analysis, and used to optimize both dot-ELISA and plate-ELISA. Both dot-ELISA and plate-ELISA using recombinant Tams1 protein exhibited comparable diagnostic performance, with a kappa value of 0.826 and similar analytical productivity ( P  = 0.6165). Dot-ELISA revealed a BTT seroprevalence of 58.4% in the cattle population, demonstrating good sensitivity (93.33%) and specificity (90%). The diagnostic performance of dot-ELISA was found to be superior to other molecular techniques, including microscopy, single PCR, and nested PCR. Dot-ELISA is also a sustainable solution in comparison to other laboratory diagnostic techniques with benefits of early diagnosis, reduced waste generation, resource efficiency, cost-effective point of care disease surveillance. With its minimal antigen requirement, Tams1 molecule based dot-ELISA is recommended as an effective tool for epidemiological studies and field surveys of BTT.

Theileria orientalis , the causal agent of oriental theileriosis, is known to cause mild disease in cattle and buffalo across the world. Recently, different genotypes of T. orientalis have emerged as pathogenic, causing high reported morbidity in cattle. This study focuses on investigating three suspected outbreaks of oriental theileriosis that resulted in fatalities among crossbred and indigenous bulls in Karnataka, India. Examination of blood smears revealed the presence of T. orientalis piroplasms within erythrocytes. The genetic characterization of T. orientalis was conducted by targeting specific markers, including the mpsp gene, p23 gene, and ribosomal DNA markers (18S rRNA gene, ITS-1, and ITS-2). Analysis based on the 18S rRNA gene unveiled the presence of both Type A and Type E genotypes of T. orientalis in the outbreaks. The mpsp gene-based analysis identified genotype 7 of T. orientalis in crossbred cows, whereas genotype 1 (Chitose B) was found to be present in indigenous bulls. Haplotype network analysis based on the mpsp gene revealed the presence of 39 distinct haplotypes within the 12 defined genotypes of T. orientalis with a high haplotype diversity of 0.9545 ± 0.017. Hematological and biochemical analysis revealed a decrease in calcium, hemoglobin levels, red blood cell counts, and phosphorus. This study constitutes the initial documentation of a clinical outbreak of oriental theileriosis in indigenous bulls with genotype 1 (Chitose 1B). Substantial epidemiological investigations are imperative to gain a comprehensive understanding of the geographical distribution of distinct genotypes and the diverse clinical manifestations of the disease across various hosts.

Ovine theileriosis is a tick-borne hemoparasitic disease caused by protozoan parasites of the genus Theileria, which poses a substantial threat to sheep and goats. The disease is prevalent in northwestern China, yet studies on the diversity of its pathogens in Qinghai Province remain limited. From 2014 to 2025, 1062 blood samples were collected from sheep in 11 counties of Qinghai Province, and the Theileria spp. were investigated by PCR targeting the 18S rRNA gene. Genetic diversity analysis was performed to assess sequence variations and phylogenetic relationships. The results show that 424 samples tested positive for Theileria spp., with an overall infection rate of 39.92%, including T. uilenbergi 26.74%, T. luwenshuni 22.41%, T. ovis 18.17%, T. capreoli 0.56%, and Theileria sp. OT3 0.09%. The co-infection rate was 22.69%. Six haplotypes were detected in both T. uilenbergi and T. ovis, and fifteen in T. luwenshuni. Hap 1 was the dominant haplotype for all three species, with respective proportions of 73.08%, 77.06% and 82.17%. Phylogenetic analysis showed the strains were most closely related to those from Hunan, Turkey, Shaanxi, etc. These findings reveal a high diversity of Theileria species in Qinghai sheep, which provides crucial epidemiological insights into the transmission dynamics of ovine theileriosis in Qinghai and vital support for developing effective control strategies.

Theileria annulata is a tick-borne hemoprotozoan parasite responsible for tropical theileriosis in the bovine population, which causes substantial economic losses to the livestock sector. The present study has investigated, characterized, and shaped epidemiologic and phylogenetic profiles of T . annulata infection in the cattle population of central Khyber Pakhtunkhwa, Pakistan. A total of 600 blood samples were collected from cattle. Microscopy and PCR ( 18S rRNA taxonomic marker) assays were performed to detect T . annulata infection in cattle from the study area. The overall relative prevalence rates of T . annulata in the examined cattle population were 12.8% (microscopy) and 23.7% (PCR). District-wise analysis (microscopy/PCR) showed that cattle from district Mardan were found more infected (16.0%/28.0%), as compared to cattle from district Charsadda (13.5%/25.5%) and district Peshawar (9.0%/17.5%). Based on host demographic and ecological parameters analysis, theileriosis was found to be higher in young, female, crossbred, freely grazing, tick-infested, and irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, tick infestation, acaricides use, and feeding method were significant risk factors (P<0.05) whereas multivariate analysis indicated that host age, gender, tick infestation, acaricidal application, and feeding method were potential risk factors (P<0.05) for tropical theileriosis in the cattle population. Phylogenetic and sequence analysis showed that T . annulata 18S rRNA isolates shared homology and phylogeny with other isolates from Asia and Europe. This study has addressed the epidemiology and phylogeny of T . annulata circulating in bovid in the study area where gaps were still present. These findings will serve as a baseline and will facilitate future large-scale epidemiological investigations on tropical theileriosis in the cattle population at a national level.

Tropical theileriosis is a tick-borne disease caused by the protozoan Theileria annulata and transmitted by numerous species of Ixodid ticks of the genus Hyalomma. The main clinical signs are fever, lymphadenopathy, and anemia responsible for heavy economic losses, including mortality, morbidity, vaccination failure, and treatment cost. Development of poor cell-mediated immunity (CMI) has been observed in the case of many bovine pathogens (bacteria, viruses, and parasites). Quantification of CMI is a prerequisite for evaluating vaccine efficacy against theileriosis caused by T. annulata. The current study evaluated the CMI in calves administered with two types of T. annulata vaccine (live attenuated and killed) . We prepared a live attenuated T. annulata vaccine by attenuation in a rabbit model and also prepared killed vaccine from non-attenuated T. annulata . For the evaluation of immune response in experimental groups including control, 20 calves were divided into four different groups (A, B, C, and D). They were either inoculated subcutaneously with live rabbit-propagated- Theileria -infected RBCs (5 × 10 6 ) (group A) or with killed T. annulata vaccine (2 × 10 9 schizonts) with Freund’s adjuvant (group B), along with an infected group (group C) and a healthy control group (group D). The protection of vaccinated calves was estimated with challenge infection. Our results showed that with a single shot of live-attenuated and killed vaccine with a booster dose elicited cell-mediated immune responses in immunized calves. We observed a significant elevation in CD4 + and CD8 + T cells in immunized calves. A significant difference in the CD8 + T cell response between the post-challenge stage of killed and live vaccine ( p  < 0.0001) was observed, whereas no other difference was found at both pre- and post-immunization stages. A similar finding was recorded for the CD4 + T cells at a post-challenge stage, where a significant difference was seen between killed and live vaccine ( p  < 0.0001). Another significant difference was observed between the CD8 + T cells and CD4 + T cells at the post-challenge stage in the live vaccine group, where there was a significantly higher induction of CD4 + T cell response ( p  < 0.0001).

Equine theileriosis, a tick-transmitted disease caused by the hemoprotozoan parasites Theileria equi and Theileria haneyi , affects equids throughout tropical and subtropical regions of the world. It is a significant regulatory concern in non-endemic countries, where testing for equine theileriosis is required prior to horse import to prevent parasite entry. Within endemic areas, infection causes significant morbidity and mortality, leading to economic losses. No vaccine for equine theileriosis is available, and current drug treatment protocols are inconsistent and associated with significant side effects. Recent work has revealed substantial genetic variability among equine theileriosis organisms, and analysis of ribosomal DNA from affected animals around the world indicates that the organisms can be grouped into five distinct clades. As these diverse parasites are capable of infecting a wide range of both tick and mammalian hosts, movement of different equine Theileria species between endemic countries, and eventually into non-endemic countries, is a significant concern. Furthermore, the substantial genetic variability of these organisms will likely render currently utilized importation diagnostic tests unable to detect all equine Theileria spp. To this end, more complete characterization of these diverse parasites is critical to the continued global control of equine theileriosis. This review discusses current knowledge of equine Theileria spp. in this context, and highlights new opportunities and challenges for workers in this field.

Control of Theileria equi, the primary cause of equine theileriosis, is largely reliant on acaracide use and chemosterilization with imidocarb dipropionate (ID). However, it is currently unknown if ID is effective against Theileria haneyi, the recently identified second causative agent of equine theileriosis, or if the drug maintains effectiveness against T. equi in the presence of T. haneyi co-infection. The purpose of this study was to address these questions using ID treatment of the following three groups of horses: (1) five T. haneyi infected horses; (2) three T. haneyi-T. equi infected horses; and (3) three T. equi-T. haneyi infected horses. Clearance was first evaluated using nPCR for each Theileria sp. on peripheral blood samples. ID failed to clear T. haneyi in all three groups of horses, and failed to clear T. equi in two of three horses in group two. For definitive confirmation of infection status, horses in groups two and three underwent splenectomy post-treatment. The T. equi-nPCR-positive horses in group two developed severe clinical signs and were euthanized. Remaining horses exhibited moderate signs consistent with T. haneyi. Our results demonstrate that ID therapy lacks efficacy against T. haneyi, and T. haneyi-T. equi co-infection may interfere with ID clearance of T. equi.

Tropical Bovine Theileriosis is an important tick-borne disease. This study aims to assess the occurrence of Theileria annulata infection in two indigenous Portuguese cattle breeds. A total of 843 blood samples collected from animals of Alentejana (n = 420) and Mertolenga (n = 423) breeds were analyzed. The detection of Theileria annulata was determined by amplification of a fragment of the merozoite-pyroplasm surface antigen gene with 319 base pairs (bp). The prevalence found (10.8%) is lower than that reported in previous studies (21.3%). A statistically significant difference was found for positivity between breeds (p < 0.05). There is also a higher probability of older animals being positive compared to younger ones (p < 0.05). The region where Mertolenga animals are located is shown to have a significant impact on positivity (p < 0.05). Thus, the development of sustainable T. annulata control strategies and their implementation, adapted to the epidemiological conditions of higher risk, will be extremely important.

Ticks are important vectors of an array of viral, bacterial and protozoan pathogens resulting in a wide range of animal and human diseases. There is limited information in the literature about tick species in the Middle East and North Africa (MENA) countries, even though they have suitable climate and vegetation for ticks and their hosts. We reviewed the occurrence of tick species and the pathogens they transmit from the MENA on published papers from 1901–2020. We found taxonomic records of 55 tick species infesting livestock representing the following eight genera: Ornithodoros, Otobius, Amblyomma, Dermacentor, Haemaphysalis, Hyalomma, Ixodes, and Rhipicephalus. In addition, 15 pathogens were recorded causing diseases of significance, with Crimean–Congo hemorrhagic fever, theileriosis, babesiosis and anaplasmosis being widely distributed diseases in the region. In recent decades, there has been increasing trends in disease occurrence and movement associated with global movement of humans and global trade of animals. We suggest that disease control and prevention could be achieved effectively through good integration between public health, veterinary medicine and animal management, and ecological approaches. We recommend further research in the areas of tick ecology and tick born-disease transmission. Furthermore, we suggest evaluation and improvement of disease control policies in the region.

Benign theileriosis, caused by the members of the Theileria orientalis complex, can develop fatal clinical outbreaks characterized by acute respiratory manifestation in stressful conditions. This report describes the molecular diagnosis and clinical management of a recently transported buffalo calf with severe Theileria buffeli infection and associated acute pneumonia. A five-month-old male buffalo calf having an inter-state travel history three days back was presented with pyrexia, anorexia, weakness, mucoid rhinorrhoea, dyspnoea and diarrhoea from the day of procurement. The history and physical examination revealed a clinical presentation similar to shipping fever. Whereas, severe parasitemia of Theileria spp. with anaemia, thrombocytopenia and granulopenia were evident on laboratory investigation. The Theileria spp. infection was confirmed by PCR method using specific primers and the authentication was made by detailed sequence analysis. The small subunit rRNA was amplified using universal apicomplexan primers and the phylogenetic analysis was carried out for further characterisation. The animal was stabilized by steroid nebulization therapy and the specific chemotherapy was instigated using buparvaquone and sulphamethoxazole-trimethoprim combination. Supportive medications like non-steroidal anti-inflammatory drugs, antihistamines, antidiarrhoeals and vitamins were provided symptomatically. The animal showed a good response to therapy and recovered from parasitemia by day 10 and the molecular clearance was later confirmed on day 70 of therapy. The present case of Theileria buffeli infected buffalo calf with acute respiratory signs points towards the possible hemoparasitic outbreaks in transport-stressed animals with the signs of shipping fever-associated syndrome. Highlights First report of a Theileria buffeli infected buffalo calf with immediate travel history manifesting acute respiratory sickness similar to shipping fever. Complete clearance of Theileria spp. organisms obtained by combination therapy of buparvaquone and sulphamethoxazole-trimethoprim. Subclinical carrier status of benign theileriosis may cause obscure production loss and unexpected life-threatening clinical flare-ups in buffaloes.