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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 .

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.

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.

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.

Ticks transmit more pathogens than any other arthropod, and one single species can transmit a large variety of bacteria and parasites. Because co-infection might be much more common than previously thought, we evaluated the prevalence of 38 known or neglected tick-borne pathogens in Ixodes ricinus ticks. Our results demonstrated that coinfection occurred in almost half of the infected ticks, and that ticks could be infected withup to five pathogens. Moreover, as it is well established that symbionts can affect pathogen transmission in arthropods, we also evaluated the prevalence of four symbiont species and demonstrated that all ticks were infected by at least one microorganism. This work highlights the co-infection phenomenon in ticks, which may have important implications for human and animal health, emphasizing the need for new diagnostic tests better adapted to tick-borne diseases. Finally, the high co-occurrence of symbionts and pathogens in ticks, reveals the necessity to also account for these interactions in the development of new alternative strategies to control ticks and tick-borne disease.

Piroplasmid parasites such as Theileria luwenshuni protozoa pose a global threat to both animal and human health. However, human theileriosis remains underexplored compared to infections caused by Plasmodium and Babesia species parasites. We investigated potential hemoparasite infections among 1,721 persons with fever, anemia, or both in Yunnan Province, China. Molecular detection identified 13 cases positive for T. luwenshuni protozoa, of which 5 patients were further confirmed by Western blot antibody analysis. We also identified 6 babesiosis cases, 3 infections with B. microti and 3 with novel Babesia spp. Subsequent vector and host investigations in the vicinity of the index cases revealed T. luwenshuni protozoa in 1 tick and 53 livestock animals. Of note, 3.3% combined vector-host samples tested positive for genetically diverse Babesia species. Our findings highlight the endemic circulation of T. luwenshuni and Babesia spp. parasites in southwest China, underscoring their importance as emerging public health concerns.

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.

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.

In this study, we sequenced the complete mitochondrial genome of Theileria velifera and compared it with other Apicomplexan parasites. The mitochondrial genome of T. velifera is a linear monomer molecule spanning 6,125 bp, and it encodes 3 protein-coding genes (PCGs): cox1 , cob , and cox3 . Besides, it contains 5 large subunit (LSU) rRNA gene fragments and terminal inverted repeats (TIR) at both ends. Moreover, the mitochondrial genomes of most Apicomplexan parasites in this study are typically around 6,000 bp in length and are linear in structure, featuring three PCGs. The start codons observed in Thaleria spp. and Babesia spp. parasites predominantly include ATN, GTN, and TTN, while the end codons are mainly TAA, TAG, and TGA. Phylogenetic analysis showed that T. velifera was closely related to T. annulata , T. parva , T. taurotragi and T. lestoquardi . The complete mitochondrial genome sequence of T. velifera was examined and compared to other Apicomplexan parasites in this study, offering fresh perspectives on the evolution and phylogenetic connections among these parasites.

Raising small ruminants is the main source of income for farmers in Pakistan especially in rural areas of Dera Ghazi Khan in Punjab. Despite having large sheep population, the prevalence of intra-erythrocytic protozoa, Theileria ( T .) lestoquardi , has never been reported from this area. This study was conducted to fill this knowledge gap and 333 blood samples of apparently healthy small ruminants (168 sheep and 165 goats) along with their epidemiological data were collected from Dera Ghazi Khan district during August till November 2022. The polymerase chain reaction (PCR) analysis amplified a 785 base pair amplicon specific for the Merozoite surface antigen ( ms 1–2 ) gene of T . lestoquardi in 2 out of the 168 (3.3%) sheep blood samples, while no goat blood sample out of 165 was found to be infected with T . lestoquardi . DNA sequencing confirmed the presence of Theileria lestoquardi in both samples and phylogenetic analysis revealed that these amplicon resembled the partial ms 1–2 gene sequences detected in small ruminants from Pakistan, India Iran and Egypt. All the studied epidemiological factors (age, sex, breed, size of herd, dogs with herd, composition of herd, size of herd and Tick burden on sheep) were not found associated with the prevalence of T . lestoquardi . In conclusion, this study reports a low prevalence of T . lestoquardi infection in the Dera Ghazi Khan District of Punjab, Pakistan. The data generated from this work will help pave the way for the prophylactic detection and control of ovine and caprine theileriosis in the region.