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Abstract Babesia spp. and Theileria spp. are tick-borne protozoan parasites with veterinary importance. In China, epidemiological and genetic investigations on many Babesia and Theileria species were still absent in many areas and many tick species. From Aug 2021 to May 2023, 645 ticks were collected from the body surface of domestic animals (camels, goats, sheep, and cattle) using tweezers in seven counties in three provinces including Xinjiang (Qitai, Mulei, Hutubi, and Shihezi counties), Chongqing (Youyang and Yunyang counties), and Qinghai (Huangzhong county). Three tick species were morphologically and molecularly identified (334 Hyalomma asiaticum from Xinjiang, 245 Rhipicephalus microplus from Chongqing, and 66 Haemaphysalis qinghaiensis from Qinghai). A total of three Babesia species and two Theileria species were detected targeting the 18S gene. The COI and cytb sequences were also recovered from Babesia strains for further identification. In R. microplus from Chongqing, Babesia bigemina, the agent of bovine babesiosis, was detected. Notably, in H. asiaticum ticks from Xinjiang, a putative novel genotype of Babesia caballi was identified (0.90%, 3/334), whose COI and cytb genes have as low as 85.82% and 90.64–90.91% nucleotide identities to currently available sequences. It is noteworthy whether the sequence differences of its cytb contribute to the drug resistance of this variant due to the involvement of cytb in the drug resistance of Babesia. In addition, Theileria orientalis and Theileria annulata were detected in R. microplus from Chongqing (12.20%, 31/245) and H. asiaticum from Xinjiang (1.50%, 5/334), respectively. These results suggest that these protozoan parasites may be circulating in domestic animals in these areas. The pathogenicity of the novel genotype of B. caballi also warrants further investigation.

Equine piroplasmosis (EP) is a global tick-borne disease of equids caused by the intraerythrocytic apicomplexan parasites Theileria equi and Babesia caballi, and the more recently discovered Theileria haneyi. These parasites can be transmitted by several tick species, including Dermacentor, Hyalomma, and Rhipicephalus, but iatrogenic and vertical transmission are also common. Clinical signs of EP include poor performance, fever, icterus, abortions, among others, and peracute or acute forms of infection are associated with high mortality in non-endemic areas. EP is a reportable disease and represents an important barrier for the international trade of horses and other equids, causing disruption of international equine sports. Tick control measures, serological and molecular diagnostic methods, and parasiticidal drugs are currently used against EP, while vaccines remain unavailable. Since most acaricides used in equids are non-environmentally friendly and linked to drug resistances, this is considered as an unsustainable approach. Imidocarb dipropionate (ID) and buparvaquone (BPQ) are currently the main drugs used to control the disease. However, while ID has several side and toxic effects and recurrent failures of treatment have been reported, BPQ is less effective in the clearance of T. equi infection and not available in some countries. Thus, novel alternative and effective therapeutics are needed. While current trade regulations require testing equids for EP before exportation, the lack of standardized PCR tests and limitations of the currently recommended serological assays entail a risk of inaccurate diagnosis. Hereby, we propose a combination of standardized PCR-based techniques and improved serological tests to diminish the risks of exporting EP-infected animals making equid international trade safer. In addition, this review discusses, based on scientific evidence, several idiosyncrasies, pitfalls and myths associated with EP, and identifies weaknesses of current methods of control and gaps of research, as initial steps toward developing novel strategies leading to control this disease. [ABSTRACT FROM AUTHOR]

Babesia spp. and Theileria spp. are tick-borne protozoan parasites with veterinary importance. In China, epidemiological and genetic investigations on many Babesia and Theileria species were still absent in many areas and many tick species. From Aug 2021 to May 2023, 645 ticks were collected from the body surface of domestic animals (camels, goats, sheep, and cattle) using tweezers in seven counties in three provinces including Xinjiang (Qitai, Mulei, Hutubi, and Shihezi counties), Chongqing (Youyang and Yunyang counties), and Qinghai (Huangzhong county). Three tick species were morphologically and molecularly identified (334 Hyalomma asiaticum from Xinjiang, 245 Rhipicephalus microplus from Chongqing, and 66 Haemaphysalis qinghaiensis from Qinghai). A total of three Babesia species and two Theileria species were detected targeting the 18S gene. The COI and cytb sequences were also recovered from Babesia strains for further identification. In R. microplus from Chongqing, Babesia bigemina, the agent of bovine babesiosis, was detected. Notably, in H. asiaticum ticks from Xinjiang, a putative novel genotype of Babesia caballi was identified (0.90%, 3/334), whose COI and cytb genes have as low as 85.82% and 90.64–90.91% nucleotide identities to currently available sequences. It is noteworthy whether the sequence differences of its cytb contribute to the drug resistance of this variant due to the involvement of cytb in the drug resistance of Babesia. In addition, Theileria orientalis and Theileria annulata were detected in R. microplus from Chongqing (12.20%, 31/245) and H. asiaticum from Xinjiang (1.50%, 5/334), respectively. These results suggest that these protozoan parasites may be circulating in domestic animals in these areas. The pathogenicity of the novel genotype of B. caballi also warrants further investigation. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis (EP) is a global worldwide infection, which can lead to the death of animals. Despite the causative agents of EP being well studied, there are no data on the distribution and genetic characteristics of EP agents in any region of Russia. In this study, blood samples from 750 horses from Novosibirsk province, Irkutsk province, and Altai region of Russian Siberia were examined for the presence of EP agents. Theileria equi and Babesia caballi were detected in all examined regions, with mean prevalence rates of 60.4% and 7.2%, respectively. The identified pathogens were genetically characterized by the 18S rRNA gene. The determined T. equi sequences were highly conserved and belonged to genotypes A and E, with genotype E being found in 88.6% of genotyped samples. In contrast to T. equi, B. caballi sequences were genetically diverse. Seven sequence variants of B. caballi were identified, and only two of them matched known sequences from the GenBank database. The determined B. caballi sequences belonged to four distinct branches within genotype A. Mixed infections with several variants of B. caballi or with T. equi and B. caballi were common. The conducted phylogenetic analysis based on all available B. caballi sequences of the 18S rRNA gene (> 900 bp) from GenBank and from this study first demonstrated the presence of five monophyletic clusters within genotype A and three clusters within genotype B. Thus, the genetic study of B. caballi from Siberia has significantly expanded the data on the genetic diversity of this pathogen. [ABSTRACT FROM AUTHOR]

Background: Piroplasmosis is a common and prevalent tick‐borne disease that affects equids. Objectives: To determine the infection and molecular characteristics of the piroplasms in donkeys from Xinjiang, northwestern China, we undertook a cross sectional study by collecting representative samples across several counties within the region. Methods: A total of 344 blood samples were collected from adult domestic donkeys from 13 counties in Xinjiang. PCR was conducted to test for T. equi and B. caballi in the blood samples based on the equine merozoite antigen‐1 (Ema‐1) gene and the 48 kDa rhoptry protein (BC48) gene, respectively. Results: Sixteen blood samples tested positive for piroplasms and the overall infection rate was 4.7% (16/344). Seven of the 13 counties were positive for piroplasms. Among the 16 piroplasm‐positive samples, 15 were singly infected with T. equi with an infection rate of 4.4% (15/344), and coinfection with T. equi and B. caballi was detected in one sample (0.3%, 1/344) from Wushi. Four T. equi sequence genotypes were identified and grouped into different branches of the evolutionary trees. Conclusion: These findings suggest that the infection rate of piroplasms is low in domestic donkeys in southern Xinjiang and that T. equi genotypes have a regional distribution. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis (EP) is caused by Theileria (T.) equi and/or Babesia (B.) caballi. The aim was to assess the percentage of positive test results for EP in horses in Europe and to identify risk factors for pathogen contact/infection. This study included results from PCR and competitive enzyme-linked immunosorbent assay testing requested by European veterinarians between 2008 and 2021. Binary bivariate logistic regression was used to analyze risk factors. A total of 4060 horses were included. PCR testing was positive in 9.7% (154/1589), serology for T. equi in 15.2% (393/2591) and for B. caballi in 6.8% (175/2578). The odds of positive serology increased by 6.8% (B. caballi, p = 0.008) and 9.5% (T. equi, p < 0.001) each year. Regionality had a statistically significant impact on PCR (Eastern p = 0.047/OR = 1.605; Southern p = 0.029/OR = 1.451; Central p = 0.007/OR = 0.617) and serological testing for T. equi (Southern p < 0.001/OR = 2.521; Central p < 0.001/OR = 0.537; Northern p = 0.003/OR = 0.462), as well as breeds on seroprevalence of B. caballi (heavy horses: p = 0.016/OR = 2.239) and T. equi (ponies: p = 0.007/OR = 0.340; warmbloods: p = 0.025/OR = 1.602). In conclusion, there was a significant geographical impact on the results of PCR and serology, consistent with known vector habitats. The rising numbers of horses tested serologically positive highlights the importance of surveillance. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis (EP) is a global tick-borne disease of equids caused by the intraerythrocytic apicomplexan parasites Theileria equi and Babesia caballi, and the more recently discovered Theileria haneyi. These parasites can be transmitted by several tick species, including Dermacentor, Hyalomma, and Rhipicephalus, but iatrogenic and vertical transmission are also common. Clinical signs of EP include poor performance, fever, icterus, abortions, among others, and peracute or acute forms of infection are associated with high mortality in non-endemic areas. EP is a reportable disease and represents an important barrier for the international trade of horses and other equids, causing disruption of international equine sports. Tick control measures, serological and molecular diagnostic methods, and parasiticidal drugs are currently used against EP, while vaccines remain unavailable. Since most acaricides used in equids are non-environmentally friendly and linked to drug resistances, this is considered as an unsustainable approach. Imidocarb dipropionate (ID) and buparvaquone (BPQ) are currently the main drugs used to control the disease. However, while ID has several side and toxic effects and recurrent failures of treatment have been reported, BPQ is less effective in the clearance of T. equi infection and not available in some countries. Thus, novel alternative and effective therapeutics are needed. While current trade regulations require testing equids for EP before exportation, the lack of standardized PCR tests and limitations of the currently recommended serological assays entail a risk of inaccurate diagnosis. Hereby, we propose a combination of standardized PCR-based techniques and improved serological tests to diminish the risks of exporting EP-infected animals making equid international trade safer. In addition, this review discusses, based on scientific evidence, several idiosyncrasies, pitfalls and myths associated with EP, and identifies weaknesses of current methods of control and gaps of research, as initial steps toward developing novel strategies leading to control this disease.

Abstract Background Piroplasmosis is a common and prevalent tick‐borne disease that affects equids. Objectives To determine the infection and molecular characteristics of the piroplasms in donkeys from Xinjiang, northwestern China, we undertook a cross sectional study by collecting representative samples across several counties within the region. Methods A total of 344 blood samples were collected from adult domestic donkeys from 13 counties in Xinjiang. PCR was conducted to test for T. equi and B. caballi in the blood samples based on the equine merozoite antigen‐1 (Ema‐1) gene and the 48 kDa rhoptry protein (BC48) gene, respectively. Results Sixteen blood samples tested positive for piroplasms and the overall infection rate was 4.7% (16/344). Seven of the 13 counties were positive for piroplasms. Among the 16 piroplasm‐positive samples, 15 were singly infected with T. equi with an infection rate of 4.4% (15/344), and coinfection with T. equi and B. caballi was detected in one sample (0.3%, 1/344) from Wushi. Four T. equi sequence genotypes were identified and grouped into different branches of the evolutionary trees. Conclusion These findings suggest that the infection rate of piroplasms is low in domestic donkeys in southern Xinjiang and that T. equi genotypes have a regional distribution.

Abstract Background Equine piroplasmosis is caused by two tick-borne protozoan parasites, Theileria equi and Babesia caballi,, which are clinically relevant in susceptible horses, donkeys, and mules. Moreover, equine piroplasmosis significantly constrains international trading and equestrian events. Rapidly diagnosing both parasites in carrier animals is essential for implementing effective control measures. Here, a rapid immunochromatographic test for the simultaneous detection of antibodies to T. equi and B. caballi was evaluated using samples from horses and donkeys collected in Greece, Israel, and Italy. The results were compared with an improved competitive enzyme-linked immunosorbent assay (cELISA) for detecting antibodies to both parasites using the same panel of samples. Methods Blood samples were collected from 255 horses and donkeys. The panel consisted of 129 horses sampled at four locations in northern Greece, 105 donkeys sampled at four locations in Sicily, and 21 horses sampled at two locations in Israel. The rapid test and the cELISA were performed according to the manufacturer’s instructions, and the results were subjected to a statistical analysis to determine the sensitivity and specificity of both tests and their association. Results The immunochromatographic test provided a result within 15 min and can be performed in the field, detecting both pathogens simultaneously. The overall coincidence rate between the rapid test and the cELISA for detecting antibodies against T. equi was 93% and 92.9% for B. caballi. The rapid test’s sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for T. equi were above 91.5%. Sixteen samples were positive for both parasites in the rapid test and eight in the cELISA. Either test had no significant association between T. equi and B. caballi detection. The detection rates of both parasites were significantly higher in Italy than in Greece or Israel and in donkeys than in horses. The agreement for T. equi between the results of both tests was high in Greece (93.8%) and Italy (95.2%) and moderate in Israel (76.2%). For B. caballi, the specificity and NPV of the rapid test were high (94.2% and 98.3%, respectively), although the sensitivity and PPV were moderate (69.2% and 39.1%, respectively) due to the small sample size. However, for B. caballi, the sensitivity was higher with the rapid test. Conclusions The rapid test detected T. equi and B. caballi simultaneously in the field, potentially replacing laborious cELISA testing and is recommended for import/export purposes. The test can also be helpful for the differential diagnosis of clinical cases, since seropositivity may rule out equine piroplasmosis since it does not indicate current or active infection. Graphical Abstract

Purpose: Piroplasmosis is responsible for anemia, fever, loss of physical activity and even death in equines. In epidemiological studies, accurate diagnostic tests are essential for detecting asymptomatic carriers. This study aimed to investigate the prevalence of infection in asymptomatic horses from Lorestan province, western Iran by developing a multiplex PCR. Methods and results: Blood samples were examined by microscopy and multiplex PCR targeting the SSU rRNA gene of Theileria equi and Babesia caballi. Out of the total of 165 horses, 19 (11.51%) and 31 (18.78%) cases were positive for piroplasms by microscopy and PCR, respectively. The detection rates of both genera were significantly higher in multiplex PCR compared to microscopy (p < 0.0001). Compared with multiplex PCR, the sensitivities of microscopy for the detection of Babesia were only 28.5%. The prevalence of T. equi infection was significantly higher in summer (p = 0.035). The prevalence of B. caballi was significantly higher in males (p = 0.038). Conclusion: Findings indicate that the multiplex PCR described here is a sensitive technique for the detection of piroplasm DNA in carriers. Furthermore, asymptomatic carriers must be considered as an important source of infection for equids living in this region. [ABSTRACT FROM AUTHOR]

The purpose of the research is the characterization of species composition and abundance of ixodid ticks that parasitize on horses, seasonal dynamics of ticks inhabiting natural pastures and their piroplasmid infection. Materials and methods. The studies were performed in four Gorny Altai physiographic provinces. Ticks were manually collected from horses (1183 specimens). Ticks were counted on natural pastures every ten days with subsequent determination of the species composition (1109 specimens). Collected ticks of the genus Dermacentor (443 specimens) were examined by nested PCR in the presence of genus-specific primers for Babesia spp. and Theileria spp.; the species identification of discovered piroplasmas was determined by 18S rRNA gene sequencing. Results and discussion. Ixodid ticks of three genera (Dermacentor, Ixodes, Haemaphysalis) and 6 species: D. nuttalli, D. silvarum, D. reticulatus, D. marginatus, H. concinna and Ix. Persulcatus, were recorded in the collections from horses and pastures. The most numerous species was D. nuttalli (56.1%), and the most common, Ix. persulcatus, that was recorded in all districts except Kosh-Agachsky in South-Eastern Altai. D. silvarum accounted for 14.9%, and other species were represented to a lesser extent. The highest species diversity, 5 species, was observed in the Northern Altai pastures and only 1 species, D. nuttalli, was recorded in the South-Eastern Altai. The seasonal dynamics of ticks in the pasture was characterized by two peaks in numbers, namely, the spring peak in the 3rd decade of April and the autumn peak in the 2nd decade of October. The spring-summer period (March-June) had 87.1% of recorded ticks, and the autumn period (September-October) had 7.2%. Three species of the examined ticks (D. silvarum, D. marginatus, D. nuttalli) were found to have DNA of three piroplasmid species (Babesia caballi, Theileria equi and Babesia sp). Piroplasmids infection rate in ticks was 2.7-25.0%, and 2.7% on average in Gorny Altai.

Abstract Background Babesia caballi is an intraerythrocytic parasite from the phylum Apicomplexa, capable of infecting equids and causing equine piroplasmosis. However, since there is limited genome information available on B. caballi, molecular mechanisms involved in host specificity and pathogenicity of this species have not been fully elucidated yet. Results Genomic DNA from a B. caballi subclone was purified and sequenced using both Illumina and Nanopore technologies. The resulting assembled sequence consisted of nine contigs with a size of 12.9 Mbp, rendering a total of 5,910 protein-coding genes. The phylogenetic tree of Apicomplexan species was reconstructed using 263 orthologous genes. We identified 481 ves1-like genes and named “ves1c”. In contrast, expansion of the major facilitator superfamily (mfs) observed in closely related B. bigemina and B. ovata species was not found in B. caballi. A set of repetitive units containing an open reading frame with a size of 297 bp was also identified. Conclusions We present a chromosome-level genome assembly of B. caballi. Our genomic data may contribute to estimating gene expansion events involving multigene families and exploring the evolution of species from this genus.

Equine piroplasmosis is reported to be mostly endemic in tropical and subtropical regions of the world including Africa. The disease is of major negative economic significance by affecting the trade of horses as well as leading to poor performance of horses and death. The study was aimed at ascertaining the prevalence of equine piroplasmosis and its associated hematological changes in horses in the Accra Metropolis. Simple random sampling technique was used to sample a total of 101 horses from the Shangrilla, Osu, and the Burma Camp. Whole blood samples were obtained from the jugular vein of all sampled horses and subjected to microscopy and complete blood count analysis. Data collected was subjected to descriptive statistics and independent t-test using Statistical Package for Social Sciences (SPSS) version 26.0. Statistical significance was tested at a 5% significance level (P-value ≤ 0.05). The overall prevalence of equine piroplasmosis was 3.96% of which Theileria spp. was the only piroplasm identified. The mixed breed of horses was most infected as compared to the foreign and indigenous breeds. The location of horses had a significant effect on the prevalence of equine piroplasmosis. With respect to the blood parameters, there was a significant reduction in the packed cell volume (PCV) for the infected horses as compared to the uninfected horses. Equine piroplasms exist in horses in Ghana; hence, further studies should be carried out in order regions to obtain a nationwide view of equine piroplasms in Ghana using serology and molecular diagnostic techniques. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis (EP) is caused by Theileria (T.) equi and/or Babesia (B.) caballi. The aim was to assess the percentage of positive test results for EP in horses in Europe and to identify risk factors for pathogen contact/infection. This study included results from PCR and competitive enzyme-linked immunosorbent assay testing requested by European veterinarians between 2008 and 2021. Binary bivariate logistic regression was used to analyze risk factors. A total of 4060 horses were included. PCR testing was positive in 9.7% (154/1589), serology for T. equi in 15.2% (393/2591) and for B. caballi in 6.8% (175/2578). The odds of positive serology increased by 6.8% (B. caballi, p = 0.008) and 9.5% (T. equi, p < 0.001) each year. Regionality had a statistically significant impact on PCR (Eastern p = 0.047/OR = 1.605; Southern p = 0.029/OR = 1.451; Central p = 0.007/OR = 0.617) and serological testing for T. equi (Southern p < 0.001/OR = 2.521; Central p < 0.001/OR = 0.537; Northern p = 0.003/OR = 0.462), as well as breeds on seroprevalence of B. caballi (heavy horses: p = 0.016/OR = 2.239) and T. equi (ponies: p = 0.007/OR = 0.340; warmbloods: p = 0.025/OR = 1.602). In conclusion, there was a significant geographical impact on the results of PCR and serology, consistent with known vector habitats. The rising numbers of horses tested serologically positive highlights the importance of surveillance.

Background: Babesia caballi is an intraerythrocytic parasite from the phylum Apicomplexa, capable of infecting equids and causing equine piroplasmosis. However, since there is limited genome information available on B. caballi, molecular mechanisms involved in host specificity and pathogenicity of this species have not been fully elucidated yet. Results: Genomic DNA from a B. caballi subclone was purified and sequenced using both Illumina and Nanopore technologies. The resulting assembled sequence consisted of nine contigs with a size of 12.9 Mbp, rendering a total of 5,910 protein-coding genes. The phylogenetic tree of Apicomplexan species was reconstructed using 263 orthologous genes. We identified 481 ves1-like genes and named "ves1c". In contrast, expansion of the major facilitator superfamily (mfs) observed in closely related B. bigemina and B. ovata species was not found in B. caballi. A set of repetitive units containing an open reading frame with a size of 297 bp was also identified. Conclusions: We present a chromosome-level genome assembly of B. caballi. Our genomic data may contribute to estimating gene expansion events involving multigene families and exploring the evolution of species from this genus. [ABSTRACT FROM AUTHOR]

Existen hemoparásitos (protozoarios y bacterianos) intracelulares que parasitan diversas células sanguíneas. También, en forma extracelular, como el protozoario Trypanosoma evansi, el cual ocasiona enfermedades en animales y humanos. En zonas tropicales, como Costa Rica, la transmisión de hemoparásitos se favorece por la abundancia de artrópodos hematófagos que actúan como vectores biológicos (por ejemplo, las garrapatas son vectores biológicos de los protozoarios Babesia sp. y Theileria sp. y bacterias como Ehrlichia sp. y Anaplasma sp.) y vectores mecánicos (moscas y tábanos son vectores mecánicos de Anaplasma marginale y Trypanosoma evansi). El objetivo de esta investigación es determinar la presencia de parásitos sanguíneos (Anaplasma spp., Ehrlichia spp., Babesia caballi, Theileria equi y Trypanosoma evansi) en los equinos de la Unidad de la Policía Montada de Costa Rica y su relación con el estado general de salud. En total, 41 equinos se sometieron a un examen clínico. Seguidamente, se tomaron muestras sanguíneas y de ectoparásitos en los equinos como en el ambiente cercano a estos. Se revisó, además, el historial clínico (padecimientos previos y resultados de hemogramas recientes). El ADN de las muestras (sangre y artrópodos) fue extraído y analizado mediante diferentes protocolos de la técnica de reacción en cadena de la polimerasa (PCR) convencional o en tiempo real. Ocho (19.5%) equinos resultaron positivos a hemoparásitos, una nueva especie de Anaplasma (n=2), coinfección de esta nueva especie de Anaplasma con B. caballi (n=2), B. caballi (n=1), T. equi (n=1), especie nueva de Ehrlichia (n=1). En un equino solamente se pudo detectar la presencia de Anaplasmataceae. Stomoxys calcitrans, recolectadas cerca de los caballos, resultaron positivas a B. caballi. Tanto el grupo de equinos PCR positivos, como PCR negativos presentaron anemia, en ausencia de signos clínicos. Se recomienda secuenciar el genoma completo de los dos patógenos nuevos detectados en los caballos (Anaplasma sp. y Ehrlichia sp.) como investigar la competencia vectorial de las moscas Stomoxys calcitrans para hemoparásitos en equinos.

The purpose of the research is to study the prophylactic efficacy of piroplasmicides and to elucidate the effect of prophylactic doses of Imidocarb Dipropionate on the pathogen viability.Materials and methods. Babezan 12% was intramuscularly injected to 150 horses based on the active substance of Imidocarb Dipropionate at 2.5 mg/kg of the body weight and Neosidin M was injected to 30 horses twice at a dose of 2.5 mL per 100 kg of the live weight with a 15-day interval. The control group of animals did not receive the drug. The clinical follow-up of the experimental and control animals’ condition was done for 72 days. Before the experiment and 14 days after the drugs, blood samples were examined by the nested PCR in the presence of genus-specific primers from the 18S rRNA gene sequence for the Babesia spp. / Theileria spp. DNA. Species identification and genotyping of the detected Piroplasma were performed by sequencing 18S rRNA gene fragments.Results and discussion. Among 12 examined horses, 8 animals (66.7%) had the Piroplasmida DNA found in the blood samples, of which 50.0% were identified as Theileria equi and 16.7% as Babesia caballi. A high T. equi DNA prevalence (over 50.0%) indicates an endemic course of equine theileriosis. Babesan 12% in early chemotherapy of the horses helped to prevent the incidence of theileriosis within 48 and 59 days. Double chemotherapy of the horses with Neosidin M with a 15-day interval prevented the morbidity for 52 days. Babesan 12% at the preventive dose had no effect on the viability of persistent T. equi stages.

Background: Equine piroplasmosis (EP) is currently not endemic in the UK, despite a lack of formal surveillance and the presence of carrier horses in the equine population. Pathogen establishment would have significant welfare and economic impacts on the national equine industry, but the disease is often overlooked by UK practitioners. Objectives: To assess the risk of disease entry, exposure and consequences to the UK equine population. Study design: Qualitative risk assessment. Methods: A qualitative risk assessment was constructed utilising the current World Organisation for Animal Health (OIE) published framework for importation risk assessment, assessing the key areas of disease entry, exposure and consequences to the UK equine population. Results: The overall risk of EP entry to the UK via importation of infected equidae with acute disease is very low but considered medium with subclinical carrier animals. Entry via importation of ticks or the importation of blood is considered very low. The risk of EP exposure to susceptible equidae in the UK is considered low by the infection routes of tick‐bites, contaminated needles and contaminated blood, but very high via transplacental transfer. However, the consequences of EP endemic establishment are considered of high significance to the UK equine industry. Main limitations: A lack of available numerical data for events and variables in disease import risk meant a qualitative assessment was the most practical method for this scenario. Conclusions: This risk assessment highlights that EP positive animals are able to enter and are currently present in the UK, and that conditions do exist that could allow forward transmission of the disease. It has highlighted a gap in existing policy where the UK falls behind OIE guidelines and has suggested steps to correct this discrepancy and improve national biosecurity. [ABSTRACT FROM AUTHOR]

Piroplasmosis is a disease that negatively affects equine health worldwide. Hence, 324 blood samples were collected from grazing horses in ten sites in Xinjiang and testing them for the presence of Theileria equi and Babesia caballi by PCR of the EMA-1 gene and BC48 gene, respectively. Of the 324 blood samples, 161 (49.7%) were positive for equine piroplasms. The prevalence of T. equi was 38.9% (126/324), while that of B. caballi was 30.2% (98/324). The T. equi and B. caballi co-infection rate was 19.4% (63/324). From the 126 EMA-1 gene sequences and 98 BC48 gene sequences we obtained, 21 and 27 genotypes were identified, respectively. The EMA-1 sequences together with the GenBank reference sequences grouped into four clusters, with those from the present study forming two distinct clusters. In contrast, the BC48 sequences formed eight clusters with the GenBank reference sequences, while those obtained in the present study formed five distinct clusters. Our results highlight the widespread distribution and abundant gene polymorphism of T. equi and B. caballi in grazing horses from Xinjiang. [ABSTRACT FROM AUTHOR]

Theileria equi (T. equi) and Babesia caballi (B. caballi) are the causative pathogens of Equine piroplasmosis (EP), a disease that has brought huge economic losses and great restrictions to the global equine industry. Rapid and accurate diagnostic methods are critical for the effective monitoring of the disease. In this study, we developed novel competitive ELISA methods and western blot assays based on the EMA1 or Bc48 proteins to detect antibodies against T. equi or B. caballi, respectively. In the novel cELISA, horseradish peroxidase (HRP)-labeled monoclonal antibodies are used in place of enzyme-conjugated secondary antibodies, in order to speed up the entire procedure. These methods have high sensitivity and no cross-reactivity with antibodies against other equine diseases. In the newly developed western blot assays, we optimized the dilution of T. equi or B. caballi positive serum samples to 1:200. Compared with the commercially available kit, both the novel cELISA assay and the western blot assay showed high coincidence rates in detecting antibodies against T. equi and B. caballi. Taken together, the novel cELISA and the western blot assays for detecting antibodies against T. equi or B. caballi have the potential to rapidly test for T. equi or B. caballi and to contribute to the surveillance and control of this disease. [ABSTRACT FROM AUTHOR]

Background Ticks in Xinjiang distribute widely and account for one third of China. Ticks can carry and transmit bacteria, virus, and parasite. However, the research of tick‐borne pathogens in Xinjiang is rather little. Objective To understand the situation of hard tick carry Theileria equi, Babesia caballi and Rickettsia spp. of Zhaosu and Altay in Xinjiang. Methods In this study, 119 tick samples were obtained from horses in Xinjiang, China, Ticks were identified morphologically to determine species and PCR was used to investigate the situation of pathogens by hard ticks. Results One hundred and seven belong to Dermacentor marginatus, five belong to D. niveus, and seven belong to D. silvarum. Theileria equi and Babesia caballi were detected in one tick and 18 ticks, respectively. However, the carrying rate of Rickettsia spp. was 51.26% (61/119). Among these, the mixed carriage rate of T. equi and Rickettsia spp. was 0.8% (1/119). The mixed carriage rate of B. caballi and Rickettsia spp. was 10.1% (12/119). Conclusion Our results revealed that hard tick can carry not only haeimoparasite but also many important zoonotic pathogens in Xinjiang, and this situation was worth heeding.

Equine piroplasmosis (EP) is a tick-borne disease affecting horses, donkeys, mules and zebras, caused by the intracellular apicomplexan protozoa Babesia caballi and Theileria equi. The geographical distribution of EP is closely related to the distribution of its vector tick species belonging to the genera of Dermacentor, Rhipicephalus and Hyalomma. Since the discovery of Dermacentor reticulatus ticks in 2007 and the first reported autochthonous cases in the South of the Netherlands in 2012, no data on the (sero)prevalence of EP in horses in the Netherlands have been reported and it remains unclear whether B. caballi and T. equi have been able to establish themselves in the Netherlands. This study aims to give an update on the current status of EP in horses in the Netherlands using data from serological tests performed in the context of export and screening of 12,881 horses from 2015 through 2020. Horses were categorized as “Dutch,” “Foreign,” or “Unknown” based on microchip number. The overall seroprevalence of EP in Dutch horses was found to be 0.5% (95% exact CI [0.4–0.7]), compared to 1.9% (95% exact CI [1.3–2.6]) in horses in the category “Foreign” and 1.7% (95% exact CI [1.2–2.3]) in horses in the category “Unknown.” In addition, the seroprevalence per country in the category “Foreign” ranged from 0% (0.95% exact CI [0–2.8]) for Ireland to 6.0% (0.95% exact CI [3.5–9.3]) for Spain. In light of the reports on the seroprevalence during the outbreak of autochthonous EP reported in 2012 and on seroprevalences of EP in other countries in Northwestern Europe, the seroprevalence of EP in horses exported from the Netherlands is very low. However, the higher seroprevalence of EP in horses from abroad warrants the need for the monitoring of EP, as tick vectors are present in the Netherlands and the import of horses from endemic areas increases the chances of EP becoming more prevalent in the Netherlands.

When studying a vector‐borne disease, an eco‐epidemiological approach is vital for a comprehensive understanding of how the pathogen circulates amongst populations. Equine piroplasmosis (EP), a tick‐borne disease caused by the protozoans Babesia caballi and Theileria equi, is endemic in the Mediterranean basin of Europe and causes both animal health and economic issues for the equine sector. With no vaccine available, defining the episystem of the disease can help to identify which components of the host‐pathogen‐vector‐environment system to target to improve preventive measures. In this systematic literature review, we collected relevant data on the eco‐epidemiology of EP in Europe. The 62 studies remaining after the selection procedure explored potential vectors, indicators of parasite circulation and putative risk factors of EP. Eight hard tick species were identified as potential vectors of one or both piroplasm species. Meta‐analyses were then conducted on prevalence and seroprevalence data in equids in European countries, demonstrating an estimated seroprevalence of 30% and 8% and prevalence of 25% and 2% for T. equi and B. caballi, respectively. Finally, herd management practices and environmental risk factors analysed in studies showed no real consensus between studies, but revealed a general trend highlighting age and exposure to ticks as risk factors, and vaccination as a protective factor. Through this study, we point out that only a few studies have focused on disease management practices and even fewer have studied the effect of environmental parameters on equid infections. Further investigation in these areas is required to better characterize the eco‐epidemiology of EP and risk factors associated with this disease. [ABSTRACT FROM AUTHOR]

This study was conducted to diagnose Babesia caballi in one-humped camels in Nineveh gover-norate, Iraq, using microscopic examination and conventional polymerase chain reaction (PCR) and to evaluate the clinical, haematological and blood biochemical parameters in camels. A total of 62 camels from local breeds, 27 years old from both sexes, reared in Nineveh Governorate, were natu-rally infected with B. caballi and 12 clinically healthy camels were used as controls based on clinical and laboratory examination. Babesia caballi was diagnosed as pear-shaped organisms, existing mainly in pairs within the erythrocytes in blood smears stained by Giemsa, with mean parasitaemia rate 8.59±0.13%. Blood smears stained by acridine orange and conventional PCR confirmed B. ca-balli diagnosis. Infected camels were found to be suffering from partial or total loss of appetite, ema-ciation, pale mucous membranes, irregular heart beats, rough hair coat, digestive disturbances, haemoglobinuria and presence of ticks on various body parts. There was considerably elevated body temperature, respiratory and heart rates among infected camels in comparison with control groups. Haematological examinations indicated a statistically significant (P

Abstract Background The emergence of tick-borne disease is increasing because of the effects of the temperature rise driven by global warming. In Turkey, 19 pathogens transmitted by ticks to humans and animals have been reported. Based on this, this study aimed to investigate tick-borne pathogens including Hepatozoon spp., Theileria spp., Babesia spp., Anaplasma spp., Borrelia spp., and Bartonella spp. in tick samples (n = 110) collected from different hosts (dogs, cats, cattle, goats, sheep, and turtles) by molecular methods. Methods To meet this objective, ticks were identified morphologically at the genus level by microscopy; after DNA isolation, each tick sample was identified at the species level using the molecular method. Involved pathogens were then investigated by PCR method. Results Seven different tick species were identified including Rhipicephalus sanguineus, R. turanicus, R. bursa, Hyalomma marginatum, H. anatolicum, H. aegyptium, and Haemaphysalis erinacei. Among the analyzed ticks, Hepatozoon spp., Theileria spp., Babesia spp., and Anaplasma spp. were detected at rates of 6.36%, 16.3%, 1.81%, and 6.36%, respectively while Borrelia spp. and Bartonella spp. were not detected. Hepatozoon spp. was detected in R. sanguineus ticks while Theileria spp., Babesia spp., and Anaplasma spp. were detected in R. turanicus and H. marginatum. According to the results of sequence analyses applied for pathogen positive samples, Hepatozoon canis, Theileria ovis, Babesia caballi, and Anaplasma ovis were identified. Conclusion Theileria ovis and Anaplasma ovis were detected for the first time to our knowledge in H. marginatum and R. turanicus collected from Turkey, respectively. Also, B. caballi was detected for the first time to our knowledge in ticks in Turkey.

Babesia caballi is an intra-erythrocytic parasite causing equine piroplasmosis. Three B. caballi genotypes (A, B, and C) have been identified based on the 18 S rRNA and rhoptry-associated protein (rap -1) gene sequences. These variant parasite genotypes compromise the diagnostic utility of the WOAH-recommended serological assays in declaring horses free of equine piroplasmosis. Although a gene encoding a spherical body protein 4 (sbp 4) has recently been identified as a potential antigen for the serological detection of B. caballi , the ability of this antigen to detect the different geographical strains has not been determined. The molecular distinction between variant B. caballi genotypes is limited and therefore we developed molecular typing assays for the rapid detection and quantification of distinct parasite genotypes. Field samples were screened for the presence of B. caballi using an established multiplex equine piroplasmosis qPCR assay. In this study, B. caballi genotype A was not detected in any field samples screened. However, phylogenetic analysis of the amplified sbp 4 and 18 S rRNA genes confirmed the phylogenetic groupings of the South African isolates into either B. caballi genotypes B or C. A multiple sequence alignment of the sbp 4 gene sequences obtained in this study together with the published sbp 4 sequences representing B. caballi genotype A, were used to identify conserved regions within the gene to design three primer pairs and three genotype-specific TaqMan minor-groove binder (MGB™) probes. The qPCR assays were shown to be specific and efficient in the detection and differentiation between B. caballi genotypes A, B, and C and could be used as a diagnostic assay to prevent the unintentional spread of variant B. caballi genotypes globally. [Display omitted] • Three B. caballi genotypes (A, B, C) occur that complicate diagnostics. • The sbp 4 gene was targeted for molecular typing qPCR assays. • Phylogenetic analysis confirmed the genotype grouping. • Conserved regions of the sbp 4 gene were identified for assay design. • qPCR assays detect and differentiate genotypes efficiently. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis (EP) is a tick-borne disease of horses, donkeys and wild equids. Chronically infected animals, which often only show reduced performance or are clinically unremarkable, constitute a potential risk for parasite spread. In Europe, currently no transboundary preventive measures for identifying infected animals or preventing their cross-border transportation are in place. In Austria, as in the neighbouring countries of Switzerland and Germany, EP has only been detected sporadically. By contrast, Austria’s eastern neighbour, Hungary, is considered endemic for EP. The geographic vicinity to Hungary and a recently described autochthonous case of EP induced the present pilot study on the serological status of horses in eastern Austria. Serum samples of 173 clinically healthy horses of different age and sex from 43 different locations in Vienna and Lower Austria, without known stays abroad within the last 12 months, were examined by cELISA. Sampling had been conducted during a prevalence study on West Nile infections in 2017. Four horses (2.3 %) aged 7–28 years from three different locations were positive for anti-T. equi antibodies. Two of the positive horses had a Hungarian passport, and two horses an Austrian passport. One of the horses with an Austrian passport, however, was born in Romania. No horse was positive for anti-B. caballi-antibodies. The results of this study show that animals infected with T. equi occur in the east of Austria. Equine practitioners should consider EP as a possible cause of febrile diseases and haemolytic anaemia, but also be aware of chronic or subclinical courses of infection and the risk of parasite transmission from inapparent carriers. In addition, EP should be monitored in areas with known occurrence of Dermacentor as a vector-competent tick genus. [ABSTRACT FROM AUTHOR]

Tick-borne diseases (TBDs), including emerging and re-emerging zoonoses, are of public health importance worldwide; however, TBDs tend to be overlooked, especially in countries with fewer resources, such as Zambia and Angola. Here, we investigated Rickettsia, Anaplasmataceae, and Apicomplexan pathogens in 59 and 96 adult ticks collected from dogs and cattle, respectively, in Shangombo, a town at the Zambia–Angola border. We detected Richkettsia africae and Rickettsia aeschilimannii in 15.6% of Amblyomma variegatum and 41.7% of Hyalomma truncatum ticks, respectively. Ehrlichia minasensis was detected in 18.8% of H. truncatum, and Candidatus Midichloria mitochondrii was determined in Hyalomma marginatum. We also detected Babesia caballi and Theileria velifera in A. variegatum ticks with a 4.4% and 6.7% prevalence, respectively. In addition, Hepatozoon canis was detected in 6.5% of Rhipicephalus lunulatus and 4.3% of Rhipicephalus sanguineus. Coinfection of R. aeshilimannii and E. minasensis were observed in 4.2% of H. truncatum. This is the first report of Ca. M. mitochondrii and E. minasensis, and the second report of B. caballi, in the country. Rickettsia africae and R. aeschlimannii are pathogenic to humans, and E. minasensis, B. caballi, T. velifera, and H. canis are pathogenic to animals. Therefore, individuals, clinicians, veterinarians, and pet owners should be aware of the distribution of these pathogens in the area. [ABSTRACT FROM AUTHOR]

Keywords: Anaplasma phagocytophilum; Babesia caballi; Theileria equi; Rickettsiales; protozoa; piroplasmosis; granulocytosis; horse EN Anaplasma phagocytophilum Babesia caballi Theileria equi Rickettsiales protozoa piroplasmosis granulocytosis horse N.PAG N.PAG 8 04/26/22 20220415 NES 220415 1. [Extracted from the article]

Babesia caballi, Theileria equi and several species of rickettsias are agents of vector-borne diseases that affect equines. The objective of this study was to detect infections by B. caballi and T. equi in horses and to identify rickettsias in horses and ticks in the western region of Cuba. Two nPCR assays were developed and standardized for the detection of B. caballi and T. equi. Blood samples from horses and ticks were collected. Identification by blood smear and molecular detection and identification of B. caballi, T. equi and Rickettsia spp. were carried out. Intraerythrocytic formations compatible with B. caballi and T. equi were observed. The nPCR showed that 25 % of the samples were positive for B. caballi, 73 % for T. equi and 20 % showed coinfection. The results were confirmed with the partial sequencing of the genes bc48 (B. caballi) and ema-1 (T. equi). The sequencing of the 18S rRNA gene of T. equi demonstrated the presence of at least two genotypes of T. equi isolates in Cuba. The real time qPCR assay and sequencing revealed the presence of Rickettsia amblyommatis in A. mixtum and Rickettsia felis in D. nitens. Conclusions: These results constitute the first piece of molecular evidence of B. caballi and T. equi in horses and the first report of R. felis in D. nitens in Cuba, which broadens the knowledge about the distribution of pathogens and the spectrum of potential vectors contributing to the strengthening of management and control programs. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis (EP) is a type of blood protozoan disease caused by tick-borne parasites, Theileria equi (T. equi), Babesia caballi (B. caballi) and Theileria haneyi. While many studies have been conducted on EP diagnosis, diagnostic methods exhibiting high sensitivity and specificity remain lacking. Therefore, nested PCR (nPCR) and duplex real-time fluorescence quantitative PCR (qPCR) that can simultaneously detect both T. equi and B. caballi causing agents were established and compared. The two techniques were used to analyze 36 horse blood samples for EP. This set of samples was also detected by a multinested PCR (mnPCR) targeting the EMA-1 gene of T. equi and the RAP-1 gene of B. caballi. By nPCR, duplex real-time fluorescence qPCR and mnPCR, infections with B. caballi were detected in 16.67% (6/36), 2.78% (1/36), 19.44% (7/36) of the horses, respectively. The T. equi prevalence was 58.33% (21/36) by the nPCR, 33.33% (12/36) by the duplex real-time fluorescence qPCR and 2.78% (1/36) by the mnPCR. The overall prevalence of infection with mixed parasites by nPCR was 5.56% (2/36), by duplex real-time fluorescence qPCR was 2.78% (1/36) and by mnPCR 0% (0/36). Results suggest that nPCR can detect T. equi and B. caballi positive samples with good specificity and sensitivity, although distinguishing between the two parasites requires an electrophoresis with 4% agarose gels. The duplex real-time fluorescence qPCR can readily distinguish between T. equi and B. caballi infection, but with low sensitivity.

Equine piroplasmosis (EP) is a tick-borne disease caused by Babesia caballi and Theileria equi that is potentially emerging in non-endemic countries. We conducted a descriptive study to investigate EP prevalence and spatial distribution in an endemic region: the Camargue and the Plain of La Crau in France. In spring 2015 and 2016, we carried out sampling at stables (total n = 46) with a history of horses presenting chronic fever or weight loss. Overall, we collected blood from 632 horses, which were also inspected for ticks; these horses had been housed in the target stables for at least 1 year. We obtained 585 ticks from these horses and described land use around the stables. Real-time PCR was employed to assess T. equi and B. caballi prevalence in the horses and in the ticks found on the horses. For the horses, T. equi and B. caballi prevalence was 68.6% and 6.3%, respectively. For the ticks found on the horses, prevalence was 28.8% for T. equi and 0.85% for B. caballi. The most common tick species were, in order of frequency, Rhipicephalus bursa, R. sanguineus sl., Hyalomma marginatum, Haemaphysalis punctata, and Dermacentor sp. Horses bearing Rhipicephalus ticks occurred in wetter zones, closer to agricultural areas, permanent crops, and ditches, as well as in drier zones, in the more northern countryside. Compared to horses bearing R. bursa, horses bearing R. sanguineus sl. more frequently occurred near the Rhone River. Prevalence of T. equi in the ticks was as follows: Hyalomma marginatum (43%), Dermacentor sp. (40%), R. bursa (33%), R. sanguineus sl. (19%), and Haemaphysalis punctata (17%). In contrast, B. caballi only occurred in Dermacentor sp. (20%) and R. bursa (1%). [ABSTRACT FROM AUTHOR]

Abstract Background The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. The endochin-like quinolones (ELQ)-300 and ELQ-316 have been proven to be safe and efficacious against related apicomplexans, such as Plasmodium spp., with ELQ-316 also being effective against Babesia microti, without showing toxicity in mammals. Methods The inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. The percentage of parasitized erythrocytes was measured by flow cytometry, and the effect of the ELQ compounds on the viability of horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by monitoring cell metabolic activity using a colorimetric assay. Results We calculated the half maximal inhibitory concentration (IC50) at 72 h, which ranged from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 among all cultured parasites tested at 72 h. None of the parasites tested were able to replicate in cultures in the presence of ELQ-300 and ELQ-316 at the maximal inhibitory concentration (IC100), which ranged from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 h. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions The compounds ELQ-300 and ELQ-316 showed significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

Abstract Background The tick-borne intra-erythrocytic apicomplexan Babesia caballi is one of the etiological agents of equine babesiosis, an economically important disease of equids in most tropical and subtropical areas of the world. Discovering candidate antigens for improved diagnostic tools and vaccines remains needed for controlling equine babesiosis. This study describes the B. caballi sbp4 (Bcsbp4) gene and protein (BcSBP4) and analyzes its antigenicity in infected equids. Methods BLAST searches of an uncurated B. caballi assembly genome using the B. bovis SBP4 as a query were carried out, followed by PCR amplification and sequencing of a newly identified BcSBP4. Characterization of this novel gene and protein was performed by bioinformatics analysis, western blots, immunofluorescence (IFA) and an in vitro neutralization test using anti SBP4 peptide antibodies. Antigenicity of recombinant BcSBP4 (rBcSBP4) was tested with sera from field animals (n = 18) using an indirect ELISA (iELISA). Results Babesia caballi genome searches using B. bovis SBP4 as a query allowed identification of a novel gene termed Bcsbp4. The Bcsbp4 gene encodes for a protein of 30.58 kDa, which is fully conserved among B. caballi isolates from USA and Egypt. Bioinformatics analysis indicates that BcSBP4 contains a signal peptide and lacks additional transmembrane domains. Expression of BcSBP4 in blood stages of B. caballi was confirmed by western blot and IFA using antibodies against synthetic peptides representing putative B-cell epitopes of BcSBP4 predicted by in silico analysis. In vitro neutralization tests using anti-BcSBP4 peptide antibodies showed a marginal, but statistically significant inhibitory effect on the infectivity of B. caballi merozoites in horse red blood cells. Sera from eight B. caballi-infected equids, but none out of ten negative equid control sera, gave a positive signal in an rBcSBP4 based iELISA. Conclusions The Bcsbp4 gene is expressed in B. caballi blood stages. The BcSBP4 protein is a potential candidate for developing a novel serological test that could detect B. caballi infection in equids in tropical and subtropical countries worldwide.

Abstract Background Equine piroplasmosis is a highly endemic protozoan disease of horses worldwide, caused by Theileria equi and Babesia caballi. While most horses in endemic areas are subclinically infected, the mechanisms leading to clinical outcome are vastly unknown. Moreover, since clinical signs of disease are not specific, and the prevalence in endemic areas is high, it is difficult to determine if equine piroplasmosis is the cause of disease. To identify possible mechanisms leading to the clinical outcome in an endemic area, we compared parasite loads and genotypes in clinically and subclinically infected horses. Methods Blood was collected from horses with clinical signs consistent with equine piroplasmosis, and from apparently healthy horses in Israel. Packed cell volume and total solids were measured. Quantitative and diagnostic polymerase chain reaction were used to identify, quantify and classify equine piroplasmosis infection. Phylogenetic analyses were used to determine the genotype of both parasites. Results For both parasites, clinical cases were associated with low mean packed cell volume and high mean parasite load (P

An apparently healthy 2½-year-old male Grant’s Zebra weighing approximately 200kg located at Nandankanan Zoological Park, Odisha, India, procured from Zoological Centre, Tel Aviv- Ramat Gan, Israel during September 2015 was noticed in a sitting position making frequent attempts to get up. The zebra was immobilised the same day with a combination of 1.96mg etorphine hydrochloride, 8.0mg of acepromazine and 40.0mg of xylazine hydrochloride to facilitate diagnosis and treatment. Clinical examinations did not reveal any signs suggestive of disease or disorder of the musculoskeletal system. Microscopic examination of blood smears stained in Giemsa’s stain revealed the presence of intra-erythrocytic inclusions, either single or pairs, suggestive of haemoprotozoans, i.e., B. caballi and/or T. equi. The zebra was administered with two divided doses of imidocarb injection @4.0mg/kg b.wt. deep intramuscularly in the neck region with supportive therapy. Progressive improvement in posture, gait, and appetite were noticed following 24h of medication. Three more doses of imidocarb were administered at 72h intervals, each time after immobilisation.

ABSTRACT Equine piroplasmosis (EP) is a serious problem in the horse industry, and controlling EP is critical for international horse trading. EP is caused by two apicomplexan protozoan parasites, Theileria equi and Babesia caballi. Rapid and accurate methods that are suitable for detecting these parasites in the field are crucial to control the infection and spread of EP. In this study, we developed a card to detect antibodies against T. equi and B. caballi based on two colloidal gold immunochromatographic strips according to the principle of the double-antigen sandwich. The proteins equi merozoite antigen 1 (EMA1) and rhoptry protein BC48 are commonly used as diagnostic antigens against T. equi and B. caballi, respectively. On the strip, the purified EMA1 or BC48 protein labeled with colloidal gold was used as the detector, and nitrocellulose membranes were coated with EMA1 or BC48 and the corresponding MAb as the test and control lines, respectively. The protocol takes 10 to 15 min and requires no specialized equipment or chemical reagents, and one test can detect two EP pathogens in one card. Specificity tests confirmed there was no cross-reactivity with sera positive for common equine pathogens. Using a commercial competitive enzyme-linked immunosorbent assay (cELISA) kit for comparison, 476 clinical samples were tested with the card. The coincidence rates were 96.43% and 97.90% for T. equi and B. caballi, respectively. The field trial feedback was uniformly positive, suggesting that this diagnostic tool may be useful for controlling the spread of T. equi and B. caballi. IMPORTANCE Equine piroplasmosis (EP), caused by Theileria equi and Babesia caballi, is an important tick-borne disease of equines that is prevalent in most parts of the world. EP is considered a reportable disease by the World Organization for Animal Health (OIE). The accurate diagnosis and differentiation of T. equi and B. caballi are very important for the prevention, control, and treatment of EP. Therefore, we developed a double-antigen sandwich colloidal gold immunochromatography assay (GICG) to detect T. equi and B. caballi. Two GICG strips were assembled side by side on one card for the detection of T. equi and B. caballi, and the two EP pathogens could be detected in one test. This method was simple, rapid, and specific for the detection of EP; therefore, compared to the previous methods, this method is more suitable for pathogen diagnosis in the field.

Introducción. Babesia caballi, Theileria equi y varias especies de rickettsias son agentes de enfermedades transmitidas por vectores que afectan a los equinos. El objetivo del presente estudio fue detectar infecciones por B. caballi y T. equi en caballos e identificar rickettsias en caballos y garrapatas en la región occidental de Cuba. Métodos. Se estandarizaron 2 ensa- yos de nPCR para la detección de B. caballi y T. equi. Se colectaron muestras de sangre de caballos, y garrapatas. Se realizó identificación por frotis sanguíneo y detección e identifica- ción molecular de B. caballi, T. equi y Rickettsia sp. Resultados. Se observaron formaciones intraeritrocíticas compatibles con B. caballi y T. equi. El nPCR mostró que el 25 % de las Anales de la Academia de Ciencias de Cuba; Vol. 12, 1 (2022): enero-abril ISSN 2304-0106 | RNPS 2308muestras fueron positivas para B. caballi, 73 % para T. equi y 20 % mostraron coinfección. Los resultados se confirmaron con la secuenciación parcial de los genes bc 48 (B. caballi) y ema-1 (T. equi). La secuenciación del gen ARNr 18S de T. equi demostró la presencia de al menos 2 genotipos de aislados de T. equi en Cuba. El ensayo de PCR en tiempo real y la secuenciación revelaron la presencia de Rickettsia amblyommatis en A. mixtum y Rickettsia felis en D. nitens. Como conclusiones estos resultados constituyen la primera evidencia molecular de B. caballi y T. equi en equinos y el primer reporte de Rickettsia felis en D. nitens en Cuba, lo que amplía el conocimiento sobre la distribución de patógenos y el espectro de vectores potenciales contribuyendo al fortalecimiento de los programas de manejo y control.

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Background: The emergence of tick-borne disease is increasing because of the effects of the temperature rise driven by global warming. In Turkey, 19 pathogens transmitted by ticks to humans and animals have been reported. Based on this, this study aimed to investigate tick-borne pathogens including Hepatozoon spp., Theileria spp., Babesia spp., Anaplasma spp., Borrelia spp., and Bartonella spp. in tick samples (n = 110) collected from different hosts (dogs, cats, cattle, goats, sheep, and turtles) by molecular methods. Methods: To meet this objective, ticks were identified morphologically at the genus level by microscopy; after DNA isolation, each tick sample was identified at the species level using the molecular method. Involved pathogens were then investigated by PCR method. Results: Seven different tick species were identified including Rhipicephalus sanguineus, R. turanicus, R. bursa, Hyalomma marginatum, H. anatolicum, H. aegyptium, and Haemaphysalis erinacei. Among the analyzed ticks, Hepatozoon spp., Theileria spp., Babesia spp., and Anaplasma spp. were detected at rates of 6.36%, 16.3%, 1.81%, and 6.36%, respectively while Borrelia spp. and Bartonella spp. were not detected. Hepatozoon spp. was detected in R. sanguineus ticks while Theileria spp., Babesia spp., and Anaplasma spp. were detected in R. turanicus and H. marginatum. According to the results of sequence analyses applied for pathogen positive samples, Hepatozoon canis, Theileria ovis, Babesia caballi, and Anaplasma ovis were identified. Conclusion: Theileria ovis and Anaplasma ovis were detected for the first time to our knowledge in H. marginatum and R. turanicus collected from Turkey, respectively. Also, B. caballi was detected for the first time to our knowledge in ticks in Turkey. [ABSTRACT FROM AUTHOR]

Equine piroplasmosis is caused by apicomplexan parasites, namely, Babesia caballi and Theileria equi, which are transmitted to equids principally through ticks. To ascertain the exposure of equines to agents of equine piroplasms, we tested serum samples collected from horses (n = 272) and donkeys (n = 170) in North-Western Nigeria for the presence of antibodies against B. caballi and T. equi using IFAT and ELISA. The seroprevalence of T. equi in the horses determined using IFAT and ELISA was 48.89% and 45.96%, respectively, while for B. caballi, it was 6.3% and 0.4%, respectively. For T. equi, the seroprevalence based on IFAT and ELISA results in donkeys was 14.1% and 2.9%, respectively, while for B. caballi, the seroprevalence was 2.4% and 0.6%, respectively, for ELISA and IFAT. Mixed infection detected in the horses using IFAT and ELISA was 5.5% and 0.4%, respectively, while no mixed infection was observed in the donkeys. The seroprevalence of T. equi was significantly (P <.0001) higher than that of B. caballi in both horses and donkeys. Comparatively, the IFAT detected a greater number of piroplasm seropositive animals than ELISA, indicating a difference in their diagnostic accuracy. Findings from this study confirm the existence of equine piroplasms in both horses and donkeys in North-Western Nigeria and highlights the need for robust and effective control measures against the disease. [ABSTRACT FROM AUTHOR]

Background: Ticks in Xinjiang distribute widely and account for one third of China. Ticks can carry and transmit bacteria, virus, and parasite. However, the research of tick‐borne pathogens in Xinjiang is rather little. Objective: To understand the situation of hard tick carry Theileria equi, Babesia caballi and Rickettsia spp. of Zhaosu and Altay in Xinjiang. Methods: In this study, 119 tick samples were obtained from horses in Xinjiang, China, Ticks were identified morphologically to determine species and PCR was used to investigate the situation of pathogens by hard ticks. Results: One hundred and seven belong to Dermacentor marginatus, five belong to D. niveus, and seven belong to D. silvarum. Theileria equi and Babesia caballi were detected in one tick and 18 ticks, respectively. However, the carrying rate of Rickettsia spp. was 51.26% (61/119). Among these, the mixed carriage rate of T. equi and Rickettsia spp. was 0.8% (1/119). The mixed carriage rate of B. caballi and Rickettsia spp. was 10.1% (12/119). Conclusion: Our results revealed that hard tick can carry not only haeimoparasite but also many important zoonotic pathogens in Xinjiang, and this situation was worth heeding. [ABSTRACT FROM AUTHOR]

Theileria equi (T. equi) and Babesia caballi (B. caballi) are the causative pathogens of Equine piroplasmosis (EP), a disease that has brought huge economic losses and great restrictions to the global equine industry. Rapid and accurate diagnostic methods are critical for the effective monitoring of the disease. In this study, we developed novel competitive ELISA methods and western blot assays based on the EMA1 or Bc48 proteins to detect antibodies against T. equi or B. caballi, respectively. In the novel cELISA, horseradish peroxidase (HRP)-labeled monoclonal antibodies are used in place of enzyme-conjugated secondary antibodies, in order to speed up the entire procedure. These methods have high sensitivity and no cross-reactivity with antibodies against other equine diseases. In the newly developed western blot assays, we optimized the dilution of T. equi or B. caballi positive serum samples to 1:200. Compared with the commercially available kit, both the novel cELISA assay and the western blot assay showed high coincidence rates in detecting antibodies against T. equi and B. caballi. Taken together, the novel cELISA and the western blot assays for detecting antibodies against T. equi or B. caballi have the potential to rapidly test for T. equi or B. caballi and to contribute to the surveillance and control of this disease.

Equine piroplasmosis (EP) is a widely spread tick-borne disease of horses, caused by the hemoparasites Theileria equi and Babesia caballi. Although most horses in endemic areas are subclinical carriers of parasites, acute or peracute disease may occur, especially when naïve horses are infected. This case report presents a characteristic case of peracute and fatal EP in a yearling filly, that was transferred from stable to pasture with first exposure to ticks. The filly was diagnosed with high parasitemia of both T. equi and B. caballi, with coinfection of both parasites within the same erythrocyte. Post mortem findings revealed multi-organ damage (spleen, kidney, lungs and muscles) and systemic bleeding. Quantitative PCR revealed high parasitemia of both parasites, consistent with acute and clinical infection in the sick filly. An epidemiological investigation including molecular analyses of blood samples was conducted on the farm, in which of thirteen horses, two were carriers of T. equi and one was a carrier of B. caballi with similar genotypes to the filly. The described case emphasizes the importance of implementation of preventive measures when transferring naïve horses to tick infected pasture in endemic areas. In addition, identifying the previously undocumented presence of co-infection within the same erythrocytes raises questions regarding the pathogenesis and cell invasion mechanisms of these parasites, with possible future therapeutic implications. [ABSTRACT FROM AUTHOR]

This study was conducted to diagnose Babesia caballi in one-humped camels in Nineveh governorate, Iraq, using microscopic examination and conventional polymerase chain reaction (PCR) and to evaluate the clinical, haematological and blood biochemical parameters in camels. A total of 62 camels from local breeds, 2-7 years old from both sexes, reared in Nineveh Governorate, were naturally infected with B. caballi and 12 clinically healthy camels were used as controls based on clinical and laboratory examination. Babesia caballi was diagnosed as pear-shaped organisms, existing mainly in pairs within the erythrocytes in blood smears stained by Giemsa, with mean parasitaemia rate 8.59±0.13%. Blood smears stained by acridine orange and conventional PCR confirmed B. caballi diagnosis. Infected camels were found to be suffering from partial or total loss of appetite, emaciation, pale mucous membranes, irregular heart beats, rough hair coat, digestive disturbances, haemoglobinuria and presence of ticks on various body parts. There was considerably elevated body temperature, respiratory and heart rates among infected camels in comparison with control groups. Haematological examinations indicated a statistically significant (P<0.05) decrease in red blood cell counts, haemoglobin and haematocrit with macrocytic hypochromic anaemia. Additionally, leukocytosis due to significant (P<0.05) rise in total leukocyte count and lymphocytes in infected camels in comparison with controls besides a significant (P<0.05) decrease in platelet count and fibrinogen were demonstrated. On the other hand, the mean platelet volume, platelet distribution width, clotting time, prothrombin time and activated partial thromboplastin time were significantly (P<0.05) elevated in infected camels vs control. Biochemical analysis revealed significant (P<0.05) rise in troponin I, creatine phosphokinase myocardial band and lactate dehydrogenase in infected camels compared to healthy animals. This study concluded that B. caballi infected camels might be endemic in Nineveh governorate, Iraq. Babesia caballi in camels causes clinical, haematological, and blood biochemical changes. [ABSTRACT FROM AUTHOR]