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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 .

Theileria annulata and T. parva are closely related protozoan parasites that cause lymphoproliferative diseases of cattle. We sequenced the genome of T. annulata and compared it with that of T. parva to understand the mechanisms underlying transformation and tropism. Despite high conservation of gene sequences and synteny, the analysis reveals unequally expanded gene families and species-specific genes. We also identify divergent families of putative secreted polypeptides that may reduce immune recognition, candidate regulators of host-cell transformation, and a Theileria-specific protein domain [frequently associated in Theileria (FAINT)] present in a large number of secreted proteins.

Pakistan has a huge sheep population (37.2 million in 2024) that is largely unexplored for the presence of vector transmitted parasites. Present study was aimed to document the prevalence of Anaplasma sp. , Anaplasma ovis , Theileria ovis and Theileria lestoquardi in sheep blood samples (N = 329) that were collected from six districts (Muzaffargarh, Rajanpur, Dera Ghazi Khan, Layyah, Taunsa and Khanewal) during August till December 2024 and to report the genetic diversity of screened pathogens. Molecular analyses revealed that the prevalence of Anaplasma sp. , Anaplasma ovis and Theileria ovis in screened sheep was 11%, 20% and 21% respectively. None of the screened sheep was Theileria lestoquardi infected. Co-infection of the screened pathogens was also observed. Presence of the detected pathogens was confirmed by DNA sequencing and subsequent BLAST analysis. Phylogenetic analysis revealed that these pathogens displayed genetic similarities with the sequences that were deposited from various countries across the globe. Prevalence of all screened pathogens varied significantly between the sampling districts. Similarly, the Anaplasma sp., Anaplasma ovis and Theileria ovis prevalence varied significantly among the sheep breeds. Anaplasma ovis infection was more common in large herds and in un-infested sheep. Theileria ovis infection was more frequent in small herds. In conclusion, we are reporting the presence of Anaplasma sp., Anaplasma ovis and Theileria ovis in Pakistani sheep that were enrolled from all six districts. Large-scale studies are recommended in various geo-climatic regions of Pakistan to confirm the genetic diversity, epidemiology and host-pathogen interactions that will contribute towards effective control of these infections among the local sheep population.

Strict control measures apply to movement of buffalo in South Africa including testing for Theileria parva, the causative agent of Corridor disease in cattle. The official test is a real-time hybridization PCR assay that amplifies the 18S rRNA V4 hyper-variable region of T. parva, T. sp. (buffalo) and T. sp. (bougasvlei). Mixed infections with the latter organisms affect diagnostic sensitivity due to PCR suppression. While the incidence of mixed infections in the Corridor disease endemic region of South Africa is significant, little information is available on the specific distribution and prevalence of T. sp. (buffalo) and T. sp. (bougasvlei). Specific real-time PCR assays were developed and a total of 1211 samples known to harbour these parasites were screened. Both parasites are widely distributed in southern Africa and the incidence of mixed infections with T. parva within the endemic region is similar (∼25–50%). However, a significant discrepancy exists in regard to mixed infections of T. sp. (buffalo) and T. sp. (bougasvlei) (∼10%). Evidence for speciation between T. sp. (buffalo) and T. sp. (bougasvlei) is supported by phylogenetic analysis of the COI gene, and their designation as different species. This suggests mutual exclusion of parasites and the possibility of hybrid sterility in cases of mixed infections.

Theileria equi has a biphasic life cycle in horses, with a period of intraleukocyte development followed by patent erythrocytic parasitemia that causes acute and sometimes fatal hemolytic disease. Unlike Theileria spp . that infect cattle (Theileria parva and Theileria annulata), the intraleukocyte stage (schizont) of Theileria equi does not cause uncontrolled host cell proliferation or other significant pathology. Nevertheless, schizont-infected leukocytes are of interest because of their potential to alter host cell function and because immune responses directed against this stage could halt infection and prevent disease. Based on cellular morphology, Theileria equi has been reported to infect lymphocytes in vivo and in vitro , but the specific phenotype of schizont-infected cells has yet to be defined. To resolve this knowledge gap in Theileria equi pathogenesis, peripheral blood mononuclear cells were infected in vitro and the phenotype of infected cells determined using flow cytometry and immunofluorescence microscopy. These experiments demonstrated that the host cell range of Theileria equi was broader than initially reported and included B lymphocytes, T lymphocytes and monocyte/macrophages. To determine if B and T lymphocytes were required to establish infection in vivo , horses affected with severe combined immunodeficiency (SCID), which lack functional B and T lymphocytes, were inoculated with Theileria equi sporozoites. SCID horses developed patent erythrocytic parasitemia, indicating that B and T lymphocytes are not necessary to complete the Theileria equi life cycle in vivo . These findings suggest that the factors mediating Theileria equi leukocyte invasion and intracytoplasmic differentiation are common to several leukocyte subsets and are less restricted than for Theileria annulata and Theileria parva . These data will greatly facilitate future investigation into the relationships between Theileria equi leukocyte tropism and pathogenesis, breed susceptibility, and strain virulence.

Piroplasmosis, a disease of domestic and wild animals, is caused by tick-borne protozoa in the genera of Theileria and Babesia. There is limited information available about the prevalence of piroplasmosis in ticks in China, and to assess the potential threat of piroplasmosis in China, we investigated the infections of ovine and bovine Babesia and Theileria species in ticks collected from cattle, yaks, sheep, horses, and camels in several regions of China where tick-borne diseases have been reported. In total, 652 ticks were collected from the animals in 6 provinces of China. Babesia spp. and Theileria spp. were detected with a PCR-RLB method and identified by sequencing. Overall, 157 ticks (24.1%) were infected with 5 Babesia and 4 Theileria species. Among tested tick samples, 134 (20.6%) were single infections with 1 of 7 piroplasm species, with Theileria annulata (118/652, 18.1%) being dominant. Only 23 (3.5%) tick samples were double or triple infected, Theileria luwenshuni and Theileria sinensis (18/652, 2.8%) were frequently observed in co-infections. Some piroplasm species were carried by ticks that were not previously reported to be vectors.

Tick-borne pathogens, particularly Babesia and Theileria species, are major threats to cattle production, causing economically significant diseases such as babesiosis and theileriosis. In this study, a real-time SYBR Green PCR assay was developed to detect Babesia and Theileria species in hard ticks ( N  = 65) and cattle blood samples ( N  = 143) from Thailand. Using primers targeting the mitochondrial cytochrome b gene for Babesia and the nuclear 18S rRNA gene for Theileria , the assay measured specific melting temperatures (Tm) for each species. The results showed distinct Tm values for Babesia bigemina (74.38 ± 0.04 °C), Babesia bovis (75.7 ± 0.06 °C), Theileria orientalis (74.61 ± 0.03 °C), Theileria sinensis (75.84 ± 0.03 °C), and Theileria annulata (74.06 ± 0.03 °C). The assay demonstrated high specificity, with a cutoff cycle threshold of < 35 cycles and a minimum detectable concentration of 10 copies/μL. Significant species differences in melting curves were confirmed using Tukey’s HSD test ( p  < 0.05). Theileria orientalis was detected in 8.4% of cattle blood samples, while T. sinensis was found in 25.9%, and B. bigemina in 0.7%. Theileria orientalis was also detected in 7.7% of tick samples, T. sinensis in 16.9%, and B. bigemina in 6.1%. The assay returned negative results for all non-target blood and tissue pathogens tested for specificity. This robust, high-throughput assay is highly effective for monitoring Babesia and Theileria infections, facilitating close surveillance and intervention efforts against tick-borne diseases in cattle. Les agents pathogènes transmis par les tiques, en particulier les espèces de Babesia et de Theileria , constituent une menace majeure pour la production bovine, provoquant des maladies économiquement importantes telles que la babésiose et la theilériose. Dans cette étude, un test de PCR SYBR Green en temps réel a été développé pour détecter les espèces de Babesia et de Theileria dans des tiques dures ( N  = 65) et des échantillons de sang de bovins ( N  = 143) en Thaïlande. À l’aide d’amorces ciblant le gène du cytochrome b mitochondrial pour Babesia et le gène de l’ARNr 18S nucléaire pour Theileria , le test a mesuré les températures de fusion (Tm) spécifiques de chaque espèce. Les résultats ont montré des valeurs de Tm distinctes pour Babesia bigemina (74,38 ± 0,04 °C), Babesia bovis (75,7 ± 0,06 °C), Theileria orientalis (74,61 ± 0,03 °C), Theileria sinensis (75,84 ± 0,03 °C) et Theileria annulata (74,06 ± 0,03 °C). Le test a démontré une spécificité élevée, avec un seuil de cycle de coupure < 35 cycles et une concentration minimale détectable de 10 copies/μL. Les différences significatives entre les espèces dans les courbes de fusion ont été confirmées à l’aide du test HSD de Tukey ( p  < 0,05). Theileria orientalis a été détecté dans 8,4 % des échantillons de sang de bovins, tandis que T. sinensis a été trouvé dans 25,9 % et B. bigemina dans 0,7 %. Theileria orientalis a également été détectée dans 7,7 % des échantillons de tiques, T. sinensis dans 16,9 % et B. bigemina dans 6,1 %. Le test a donné des résultats négatifs pour tous les agents pathogènes sanguins et tissulaires non ciblés testés pour la spécificité. Ce test robuste et à haut débit est très efficace pour le suivi des infections à Babesia et Theileria , facilitant ainsi la surveillance étroite et les interventions contre les maladies transmises par les tiques chez les bovins.

Piroplasmosis is an important tick-borne disease in several regions, and can lead to significant economic animal production losses. The current study aimed to systematically examine the incidence of bovine piroplasmosis in Kashgar, Xinjiang, to provide baseline data for the effective prevention and control of this disease among bovines in the region. A total of 1403 bovine blood samples from 12 sampling points were screened via PCR with universal Piroplasma primers targeting the 18S rRNA locus and specific Theileria annulata primers targeting the cytochrome b ( COB ) gene. The overall prevalence of bovine Piroplasma was 65.9% (925/1403). Three species of pathogenic Theileria , including T. annulata , T. orientalis , and T. sinensis , were detected, and the infection rates for these species were 65.1% (913/1403), 0.5% (7/1403), and 0.1% (1/1403), respectively. The mixed infection rate for T. orientalis and T. annulata was 0.3% (4/1403). No Babesia was detected in this study. In conclusion, bovine piroplasmosis was still common in Kashgar and T. annulata was the dominant species, and a mixed infection of T. annulata and T. orientalis was detected. Notably, T. sinensis was reported for the first time in this region. Therefore, strategies for the prevention and control of bovine piroplasmosis should be enhanced.

Parasites of the Theileria genus infect cattle and transform their host cells, a transformation that can be reversed by treatment with the drug buparvaquone; here, a Theileria homologue of the peptidyl-prolyl isomerase PIN1 is shown to be secreted into the host cell, where it promotes transformation and can be directly inhibited by buparvaquone. Link between theileriosis and cancer Protozoan parasites of the Theileria genus are pathogenic in cattle. Uniquely among eukaryotic parasites, Theileria can transform host leukocytes to produce proliferative and invasive phenotypes associated with JNK and AP-1 signalling pathways. The transformation can be reversed by the antitheilerial drug buparvaquone. Here Justine Marsolier et al . identify a homologue of the peptidyl-prolyl isomerase PIN1 in Theileria annulata (TaPIN1), showing that it is secreted into the host cell where it interacts with the host's ubiquitin ligase FBW7, resulting in the stabilization of c-JUN which promotes transformation. The authors also show that TaPIN1 is directly inhibited by buparvaquone, using in vitro and in vivo zebrafish xenograft experiments, and demonstrate that TaPIN1 is mutated in a drug-resistant strain. This work highlights a surprising link between parasites and host oncogenic pathways. Infectious agents develop intricate mechanisms to interact with host cell pathways and hijack their genetic and epigenetic machinery to change host cell phenotypic states. Among the Apicomplexa phylum of obligate intracellular parasites, which cause veterinary and human diseases, Theileria is the only genus that transforms its mammalian host cells 1 . Theileria infection of bovine leukocytes induces proliferative and invasive phenotypes associated with activated signalling pathways, notably JNK and AP-1 (ref. 2 ). The transformed phenotypes are reversed by treatment with the theilericidal drug buparvaquone 3 . We used comparative genomics to identify a homologue of the peptidyl-prolyl isomerase PIN1 in T. annulata (TaPIN1) that is secreted into the host cell and modulates oncogenic signalling pathways. Here we show that TaPIN1 is a bona fide prolyl isomerase and that it interacts with the host ubiquitin ligase FBW7, leading to its degradation and subsequent stabilization of c-JUN, which promotes transformation. We performed in vitro and in silico analysis and in vivo zebrafish xenograft experiments to demonstrate that TaPIN1 is directly inhibited by the anti-parasite drug buparvaquone (and other known PIN1 inhibitors) and is mutated in a drug-resistant strain. Prolyl isomerization is thus a conserved mechanism that is important in cancer and is used by Theileria parasites to manipulate host oncogenic signalling.

Theileria and Babesia are emerging threats to wildlife health but remain underreported in captive large herbivores. This study aimed to investigate the presence and genetic identity of Theileria and Babesia in large captive herbivores in Thailand using PCR targeting the 18 S rRNA gene. Blood samples were collected from 31 individuals representing five herbivore species: Malayan tapirs ( Tapirus indicus ), white rhinoceroses ( Ceratotherium simum ), pygmy hippopotamuses ( Choeropsis liberiensis ), bantengs ( Bos javanicus ), and gaurs ( Bos gaurus ) across five zoological parks in central Thailand. A total of 16 positive samples were identified, including one coinfection, resulting in an overall infection rate of 51.6% (16/31; 95% CI: 33.1–69.9). Theileria equi -like was detected in 37.5% (3/8; 95% CI: 8.5–75.5) of Malayan tapirs. Theileria bicornis was detected in 75% (9/12; 95% CI: 42.8–94.5) of white rhinoceroses. In gaur, the infection rate was 33.3% (4/12; 95% CI: 9.9–65.1), comprising one Babesia ovata infection, two Theileria orientalis , and one coinfection. This study provided the first molecular confirmation of Babesia ovata infection in gaurs. No infections were detected in pygmy hippopotamuses or bantengs. These results provide novel baseline data on tick-borne pathogens in captive environments, highlighting potential risks to susceptible wildlife, both non-domestic and domestic species, and underscoring implications for conservation. Our findings emphasize the need for continued surveillance, integrated vector management, and targeted control strategies in zoological settings to mitigate pathogen transmission and protect animal health.