Piroplasm Infection Research Articles

First Molecular Evidence of Babesia caballi and Theileria equi in Imported Donkeys from Kyrgyzstan.

Equine piroplasmosis (EP) is an important tick-borne disease of equids, caused by Theileria equi, Theileria haneyi, and Babesia caballi. Nonetheless, there has been a scarcity of systematic reports on EP parasites in donkeys in Kyrgyzstan, Central Asia. In this study, piroplasms were screened in 1900 blood samples from imported donkeys from the Osh Oblast (southwestern Kyrgyzstan) by targeting partial 18S ribosomal RNA using the polymerase chain reaction (PCR). Through molecular and phylogenetic analyses, all positive samples were sequenced to identify the species and genotypes. The results indicated the presence of both B. caballi and T. equi, with prevalence rates of 8.4% (160/1900) and 12.2% (232/1900), respectively. By amplifying part of the Erythrocyte Merozoite Antigen 1 (EMA-1) and Rhoptry-Associated Protein (RAP-1) genes, B. caballi genotype B and T. equi genotype A were identified. To the best of our knowledge, this is the first report on piroplasm infection among donkeys from Kyrgyzstan.

Molecular occurrence and genetic identification of Babesia spp. and Theileria spp. in naturally infected cattle from Thailand.

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

Are Colpodella Species Pathogenic? Nutrient Uptake and Approaches to Diagnose Infections.

Colpodella species are free-living protists phylogenetically related to apicomplexans. Colpodella sp. have been detected in human and animal tissues, as well as in ticks and biting flies. The trophozoite and cyst stages of Colpodella species can be distinguished from stages of the prey Parabodo caudatus using Sam-Yellowe's trichrome staining. Colpodella species obtain nutrients by attaching to their prey, aspirating the prey's cytoplasmic contents into a posterior food vacuole and encysting. It is unclear whether both trophozoite and cyst stages are present in human and animal tissues. Molecular techniques have detected Colpodella species in human blood, cerebrospinal fluid, and in ticks and flies. However, no morphological information was reported to aid life-cycle stage identification of Colpodella species. This review discusses the increased reports of Colpodella species detection in animals and in arthropods and the need to identify stages present in human and animal tissues. We previously used Sam-Yellowe's trichrome staining to identify life-cycle stages of Colpodella sp. In this review, we examine the reports of Colpodella species detection in human and animal tissues to determine whether the identification of Colpodella species represents true infections or contaminations of samples collected during routine surveillance of piroplasm infections in animals and arthropods. This review also aims to provide insights regarding Colpodella, nutrient uptake, and the survival of Colpodella sp. within humans, animals, and arthropods, as well as whether the attachment of trophozoites to cells occurs in tissues leading to myzocytosis and endocytosis.

Molecular epidemiological investigation of piroplasms carried by pet cats and dogs in an animal hospital in Guiyang, China.

Piroplasmosis is a zoonotic disease mainly caused by the Babesia and Theileria parasites. Piroplasmosis is often a subclinical infection in dogs and cats that is difficult to detect and is often suspected when clinical signs such as anemia are present. It has been reported to be prevalent in China. However, molecular evidence of the disease has not been reported in pet dogs and cats in Guiyang. In this study, we collected 307 anticoagulated blood samples from an animal hospital in the Wudang District of Guiyang during the period March 2021 to November 2021 and extracted DNA from the samples. The 18S rDNA gene was amplified using PCR, and the positive amplification product was sequenced. The sequences were then analyzed for homology and phylogeny. Of the 307 samples collected, 164 were feline and 143 were canine, with a total of 23 amplifying a target band of approximately 400 bp. The percentage of positives of piroplasms infection in pet cats was 4.27% (7/164), with the pathogens being T. uilenbergi (3) and T. luwenshuni (4). One Colpodella sp. and two undetermined species were also detected in the cat samples. The percentage of positives of piroplasms infection in pet dogs was 7.69% (11/143), with the pathogen being T. uilenbergi (11). One Colpodella sp. was also detected in the dog samples. The results confirmed that T. uilenbergi and T. luwenshuni are prevalent in pet cats and dogs in this area. In addition, the study found a rare zoonotic pathogen, Colpodella sp., in cats and dogs. Therefore, this study is expected to serve as a valuable reference for decision-making regarding animal health management and public health work.

Molecular detection and characterisation of Theileria in hard ticks of small ruminants in Zarrin Dasht County, Southern Iran.

Ticks are important ectoparasites of small ruminants in tropics and subtropics including Iran. They transmit serious zoonotic pathogens such as Babesia and Theileria. These parasites cause major burden on small ruminants jeopardising livelihoods of rural people in Zarrin Dasht County. This study was carried out to investigate the diversity and distribution of hard ticks of small ruminants and their piroplasm infection in a bid to contribute to Theileria and/or Babesia detection and control in Zarrin Dasht County of Fars province, Iran. We examined 751 sheep and goats from 10 sites of the County during four seasons for hard tick infestation. The collected hard ticks (994) were taxonomically identified before being separately confined in microtubes coded to indicate their species and host animals as well as site and date of collection. In total 50 pooled samples were analysed by PCR technique for Theileria and Babesia infection. The identified ticks included Hyalomma marginatum 994/362); 36.4%), Rhipicephalus turanicus 994/352); 35.51%), Hyalomma anatolicum 994/264); 26.6%), Hyalomma dromedarii 994/14); 1.41%) and Hyalomma asiaticum 994/2) 0.2%). Molecular analyses showed that 7 out of 50 pooled sample were infected with piroplasm genome in ticks shared by Theileria ovis (6:50) and Theileria lestoquardi (1:50). Babesia was absent in collected hard ticks. This is the first report on the presence of piroplasm infection in hard ticks of small ruminants in Zarrin Dasht County. Theileria ovis was more prevalent than Theileria lestoquardi but Babesia was absent. Piroplasm infection was detected in Hyalomma marginatum, Hyalomma anatolicum and Rhipicephalus turanicus. Hyalomma marginatum appears to be more competent to vector Theileria spp. This study may contribute to risk assessment and prevention of epizootic theileriosis in the County.

Piroplasm Infection in Domestic Cats in the Mountainous Region of Rio de Janeiro, Brazil.

Piroplasm infections in domestic cats have been reported with increasing frequency in numerous countries. However, in some states of Brazil, little information is available about the occurrence of these parasites. Blood samples were collected from 250 cats treated at a private clinic in the mountainous region of Rio de Janeiro. The samples were each subjected to a blood count, microscopic examination, and molecular research on piroplasms. The animals’ clinical histories and epidemiological information were analyzed to identify the risk factors associated with infection. Ticks were recovered during the clinical care and were identified and subjected to molecular analyses to determine the presence of piroplasm DNA. Piroplasms were detected in 2.7% (7/250) of the cats. Nucleotide sequences of Babesia vogeli were identified in six cats, while the Cytauxzoon sp. was identified in one cat. Cats displaying apathy/weakness/prostration and hemorrhage/bleeding were more likely to be infected. In addition, Amblyomma aureolatum was recovered from a cat PCR-negative for piroplasms. This is the first study in Rio de Janeiro that has detected Babesia vogeli in cats. The results obtained here underscore the need for further studies in Rio de Janeiro to investigate the dynamics of such infections and the vectors involved.

Genome variation in tick infestation and cryptic divergence in Tunisian indigenous sheep

BackgroundTicks are obligate haematophagous ectoparasites considered second to mosquitos as vectors and reservoirs of multiple pathogens of global concern. Individual variation in tick infestation has been reported in indigenous sheep, but its genetic control remains unknown.ResultsHere, we report 397 genome-wide signatures of selection overlapping 991 genes from the analysis, using ROH, LR-GWAS, XP-EHH, and FST, of 600 K SNP genotype data from 165 Tunisian sheep showing high and low levels of tick infestations and piroplasm infections. We consider 45 signatures that are detected by consensus results of at least two methods as high-confidence selection regions. These spanned 104 genes which included immune system function genes, solute carriers and chemokine receptor. One region spanned STX5, that has been associated with tick resistance in cattle, implicating it as a prime candidate in sheep. We also observed RAB6B and TF in a high confidence candidate region that has been associated with growth traits suggesting natural selection is enhancing growth and developmental stability under tick challenge. The analysis also revealed fine-scale genome structure indicative of cryptic divergence in Tunisian sheep.ConclusionsOur findings provide a genomic reference that can enhance the understanding of the genetic architecture of tick resistance and cryptic divergence in indigenous African sheep.

Identification of Theileria spp. and investigation of hematological profiles of their infections in goats in Hainan Island, China.

Theileria spp. are a group of parasites primarily transmitted by ticks and can pose a significant threat to domestic and wild animals globally. The main objective of this study was to understand the epidemiology of Theileria spp. in goats of Hainan Island/province, which is the only tropical region of China, and to study their hematological profiles in naturally infected goats. A total of 464 blood samples were collected from randomly selected local adult goats (Capra hircus, local domestic breed with black hair), from six cities and eight counties of Hainan, from November 2017 to October 2020. Blood smear microscopy of the sample and a nested polymerase chain reaction (nPCR) targeting the 18S rRNA gene combined with DNA sequencing were used to detect piroplasm infections in goats. Data analysis of the obtained sequences revealed that all the sequences were highly similar to the Theileria luwenshuni 18S rRNA gene sequence from the database. This result is consistent with the microscopic examination. In the hematological test, hematocrit, mean corpuscular volume, and mean corpuscular hemoglobin of the goats naturally infected with T. luwenshuni significantly increased, while mean corpuscular hemoglobin concentration and red blood cell distribution width (RDW) were significantly decreased. Results showed that T. luwenshuni could cause macrocytic, hypochromic anemia in goats. This study provides reliable and comprehensive information about the epidemiology of the parasite infections and hematological profile of the infected goats in Hainan, which encourages further investigations to develop practical control strategies for Theileria spp. infections in tropical areas.

A Survey of Tick Infestation and Tick-Borne Piroplasm Infection of Cattle in Oudalan and Séno Provinces, Northern Burkina Faso.

In this study, cattle farms located in Oudalan and Séno, two provinces in the Sahel region, northern Burkina Faso, were surveyed. Cattle owners were interviewed, cattle were examined for tick infestation, and ticks as well as blood samples were collected during the dry season (October). Blood DNA samples were tested for Babesia and Theileria infections using nested PCRs and sequencing. A total of 22 herds, 174 Zebu cattle were investigated at 6 different sites. Overall, 76 cattle (43.7 %) from 18 farms (81.8%) were found infested with ticks. Cattle in Séno, adult cattle (>5 years) and those owned by the Fulani ethnic group were significantly (p < 0.05) more likely to be tick-infested. A total of 144 adult ticks belonging to five species namely: Hyalomma impeltatum, Hyalomma impressum, Hyalomma rufipes, Rhipicephalus evertsi evertsi, and Rhipicephalus guilhoni were collected from the animals. Piroplasms were detected in the blood DNA of 23 (13.2%) cattle. The cattle in Séno and adult cattle were significantly more likely to be piroplasm-positive. Five pathogens diversely distributed were identified. Theileria mutans (12/174), Babesia bigemina (5/174), Theileria annulata (3/174), and Theileria velifera (3/174) were detected for the first time in northern Burkina Faso, whereas Babesia occultans (1/174) was found for the first time in cattle in West Africa. The analysis of the sequences, including B. bigemina RAP-1a, T. annulata Tams1 genes, and the 18S rRNA genes of all the five protozoa, revealed identities ranging from 98.4 to 100% with previously published sequences. Phylogenetic analysis based on the 18S rRNA gene sequences located north Burkina Faso piroplasms in the same clade as isolates from Africa and other regions of the world. Notably, T. mutans sequences were distributed in two clades: the T. mutans Intona strain clade and the Theileria sp. (strain MSD)/ Theileria sp. B15a clade, suggesting the presence of at least two strains in the area. These findings indicate that the control of ticks and tick-borne diseases should be taken into account in strategies to improve animal health in the Sahel region.

Molecular Characterization and Phylogenetic Analysis of Pathogenic Theileria spp. Isolated from Cattle and Sheep based on Cytochrome b Gene in Iran.

The present study investigated the phylogenetic relationship based on cytochrome b gene sequences among pathogenic Theileria species (spp.) in Iran, including Theileria annulata and Theileria lestoquardi, along with other data available in GenBank. A total of 136 (cattle) and 80 (sheep) blood samples suspected of piroplasm infection were obtained from six different provinces of Iran. Both microscopic and molecular methods using species-specific primers were used for screening T. annulata and T. lestoquardi positive samples. Finally, the partial cytochrome b gene of 30 T. annulata and 5 T .lestoquardi were amplified, sequenced, and deposited in GenBank. The results indicated that there were 12 different genotypes among T. annulata isolates, while only one genotype was observed among T. lestoquardi isolates. T. lestoquardi infection in cattle was detected in one sample, and no T. annulata and T. lestoquardi coinfection were detected in sheep and cattle. In the phylogenetic tree, different Theileria spp. were placed in separate clades, and the reliability of depicted tree and monophyly of T. annulata and T. lestoquardi ingroups were supported by the bootstrap value of 94% which significantly indicated that these two species evolved from a common ancestor. The tree also showed that these two pathogenic spp. shared a more recent common ancestor, compared to another species of Theileria parasites. To the best of our knowledge, this study is the first phylogenic analysis of pathogenic Theileria spp. in Iran based on the cytochrome b gene sequences. In addition, the first T. lestoquardi cytochrome b gene was sequenced and deposited in GenBank.

Detection of Babesia caballi, Theileria mutans and Th. velifera in ixodid ticks collected from cattle in Guinea in 2017-2018.

Intraerythrocytic protozoan parasites from the genera Babesia and Theileria may infect a wide range of animals and humans. The purpose of this study was to detect the 18S ribosomal RNA gene of Babesia spp. and Theileria spp. in ticks collected from household cows in the Republic of Guinea from 2017 to 2018 by PCR and then genotype the gene fragments by sequencing. A total of 907 ticks from 319 cows were collected in seven prefectures of Guinea (Boke, Faranah, Kankan, Kindia, Labe, Mamou and N'Zerekore). The following tick species on cattle were identified: Amblyomma variegatum (44.2%), Rhipicephalus decoloratus (34.7%), Rh. annulatus (10.3%), Rh. geigyi (7.3%) Hyalomma truncatum (2.4%), Rh. senegalensis (0.8%) and Haemaphysalis leachi (0.6%). Genetic markers for piroplasms were found in Am. variegatum, Rh. decoloratus, Rh. annulatus, and Rh. geigyi ticks, and the total infection rate for these ticks was 4.2%. The highest infection rate was found in Rh. annulatus ticks (10.9%). The piroplasms were genotyped as Babesia caballi, Theileria mutans and Theileria velifera by phylogenetic analysis of the 1150bp 18S ribosomal RNA gene fragments. These pathogens were discovered in practically all studied prefectures in Guinea except for Mamou Prefecture. We propose that these ixodid ticks might play a major role in the transmission of piroplasm infections in domestic animals in Guinea.

Molecular Detection and Identification of Piroplasms in Semi-Intensively Managed Cattle from Abeokuta, Nigeria

Abstract Piroplasmosis is a tick-borne haemolytic disease caused by different species of the Babesia and Theileria genera. Data on the prevalence of bovine piroplasms and their genetic diversity are scanty in Nigeria. Hence, this study reported the detection of some piroplasms in the blood of cattle in Abeokuta, Nigeria by the polymerase chain reaction (PCR). Blood samples were collected from 252 cattle and subjected to DNA extraction followed by PCR amplification of the partial region of 18S rRNA of the haemoprotozoans. Selected positive amplicons were unidirectionally sequenced and compared to the reference sequences from the Genbank. A total of 220 (87.3 %) cattle were positive for Theileria velifera and/or Babesia bigemina. The T. velifera was detected only in 163 (64.7 %) cattle, while 7 (2.8 %) cattle had a single infection with B. bigemina. Fifty cattle (19.8 %) had mixed infections with both parasites. There were no significant differences in piroplasm infections between the ages of cattle for both parasites. There were no significant differences in infection rates between the sexes for T. velifera, while the males had a significantly higher (P &lt; 0.05) rate of infection for B. bigemina than the female cattle. The molecular detection of Babesia and Theileria species of cattle are reported for the first time in cattle in Abeokuta, Nigeria. This study, which confirmed the endemic nature of the parasites in cattle in the study area, stresses their importance in livestock health and production in Nigeria.

Occurrence of Trypanosoma spp. and piroplasm infections of horses at Obollo-Afor southeastern Nigeria and resistance profiles of trypanosomes to isometamidium and diminazene salts

Epidemiological data on trypanosomosis and piroplasmosis of horses are lacking in southeastern Nigeria. The prevalence of trypanosome and piroplasm infections in horses and resistance profile of isolated trypanosomes to diminazene and isometamidium salts were investigated. For the cross-sectional study of horses billed for slaughter, 304 horses were randomly sampled. Approximately 2ml of blood was collected into anticoagulant-treated bottles for haematocrit (HCT) determination, direct microscopic examinations, and rat inoculation. Gender, body condition scores (BCS), age groups, and body weights of sampled horses were noted. Two isolates of Trypanosoma brucei recovered from the cross-sectional study were profiled for resistance to isometamidium hydrochloride and diminazene diaceturate in 36 BALB/c mice. Standardized protocols were used (Eisler et al., Veterinary Parasitology 97:171-182, 2001). 19.1% of horses (95% confidence interval 14.7-23.5%) were positive for haemoparasite infections including Theileria equi (16.1%) and Babesia caballi (3.9%). Only two (0.66%) Trypanosoma brucei infections were seen, being from active cases. Associations between age or gender, and presence of haemoparasites were only random. Haemoparasite-infected horses had significantly (p < 0.05) lower mean HCT and body weights and poorer BCS. From resistance profiling, for each isolate, all mice in control groups were parasitaemic by day 6 post-inoculation, while mice in test groups remained aparasitaemic over 60-day observation period. The study showed the endemicity and weights of Trypanososma spp. and piroplasm infections and among horses within the area. Furthermore, circulating strains of Trypanosoma brucei in the area are still susceptible to isometamidium and diminazene salts in mice. The pharmacoepidemiological significances of these findings were discussed.

First report of genetic diversity and risk factor analysis of equine piroplasm infection in equids in Jilin, China

BackgroundEquine piroplasmosis (EP) is a tick-borne hemoprotozoan disease of equids, caused by Theileria equi and Babesia caballi. Equine piroplasmosis represents a serious challenge to the equine industry due to important economic losses worldwide. The present study aimed to evaluate the prevalence of Theileria equi and Babesia caballi infections in equids from Jilin Province, China.MethodsA total of 220 blood samples (192 horses and 28 donkeys/mules) were collected from March 2018 to October 2019 in five districts of Jilin Province and analyzed by PCR. Potential risk factors, including the region, sex, management, and host species of the animals were assessed in relation to T. equi infection. Moreover, the V4 hypervariable region of the T. equi 18S rRNA gene was analyzed to identify specific genotypes.ResultsThe overall prevalence of T. equi in equids was 27.7%, whereas B. caballi infection was not identified. The district with the highest positive rate was Baicheng (43.3%), followed by Tonghua (28.9%), Yanbian (26.4%), Jilin (23.3%), and Liaoyuan (20.9%). The sex of the animals and farm management were identified as main risk factors, which were significantly associated with the prevalence of Equine piroplasmosis (P < 0.05). The risk factor analysis indicated that the females were at a higher risk (OR: 2.48, 95% CI: 1.17–5.27) of being infected compared to the males, whereas the organized farm was protective factor (OR: 0.42, 95% CI: 0.22–0.80). The phylogenetic analyses revealed that there were two T. equi genotypes (A and E) in Jilin.ConclusionsOur findings provided important epidemiological data for the prevention and control of T. equi infection in Jilin, China.

Molecular evidence of piroplasm infection in companion animals in Hunan Province, China

BackgroundFeline and canine babesiosis is an important tick-borne disease caused by parasites of the genus Babesia. The disease has a worldwide distribution and causes serious health problems in domestic and wild canidae and felidae.ResultsGenomic DNA was isolated from blood samples, which were randomly collected from pet dogs (n = 115) and cats (n = 25) in Changsha city of Hunan Province, China. Results of nested PCR assay targeting 18S rRNA gene and partial gene sequencing revealed that seven animals were infected with Babesia species, five dogs (5/115, 4.3%) and two cats (2/25, 8.0%). Sequence analysis showed that four dogs (3.5%) were positive for Babesia canis, and the other one for Babesia vogeli (0.87%). The two cats were infected by Babesia hongkongensis.ConclusionsThe findings of this study will expand knowledge of the distribution of Babesia species and provide important epidemiological information for the control of animal babesiosis in China.

Theileria orientalis Ikeda in host-seeking Haemaphysalis longicornis in Virginia, U.S.A.

The Asian longhorned tick, Haemaphysalis longicornis, has recently become established in the United States. In East Asia, Australia, and New Zealand, the native and previously introduced ranges, this tick is a vector of an important pathogen of cattle, Theileria orientalis. In 2017, the pathogenic Ikeda genotype of T. orientalis was associated with cattle mortalities in Virginia and in 2018 the exotic H. longicornis was detected at this same site. To investigate the possible role of this exotic tick in the epidemiology of theileriosis in Virginia, we tested host-seeking H. longicornis for piroplasm infections. We document the detection of exotic Theileria orientalis Ikeda genotype in 12.7 % (15/118) environmentally collected H. longicornis using both the 18S rRNA and major piroplasm surface protein (MPSP) gene targets. This is the first detection of a pathogen in H. longicornis in its introduced range in the United States and offers new insight into the animal health risks associated with the introduction of this exotic tick species to North America.

Detection of novel piroplasmid species and Babesia microti and Theileria orientalis genotypes in hard ticks from Tengchong County, Southwest China

To reveal the genetic diversity of Babesia microti and Theileria orientalis in Southwest China, we conducted a molecular survey of piroplasms in hard ticks in a China-Myanmar border county. Host infesting and questing ticks were collected from Tengchong County in 2013 and 2014. Piroplasm infection in ticks was detected by PCR, and then, phylogenetic analysis was conducted to study the genetic diversity of the pathogens identified in ticks. All in all, six piroplasm species comprising of B. microti; B. orientalis; a novel Babesia species designated Babesia sp. Tengchong, China; T. orientalis; T. luwenshuni; and an as yet undescribed piroplasmid species referred to as Piroplasmid sp. Tengchong, China, have been identified after screening goat- and cattle-attached ticks. In addition, B. bigemina has been identified by screening questing ticks. Phylogenetic analysis based on the 18S rRNA and partial β-tubulin gene revealed two novel potentially zoonotic genotypes designated B. microti Tengchong-Type A and B. The T. orientalis genotypes identified in the present study represent the seven known genotypes 1-5, 7, and N3 as revealed by phylogenetic analysis of 18S rRNA and MPSP genes. Importantly, an additional genotype designated N4 has also been identified in this study, which brings the number of recognized T. orientalis genotypes to a total of twelve. Thus, besides the two novel species, Babesia sp. Tengchong, China, closely related to Babesia species isolated from yak and Piroplasmid sp. Tengchong, China, our study demonstrates that additional novel B. microti and T. orientalis genotypes exist in Southwest China.

Prevalence, distribution, and diversity of cryptic piroplasm infections in raccoons from selected areas of the United States and Canada.

The order Piroplasmida contains a diverse group of intracellular parasites, many of which can cause significant disease in humans, domestic animals, and wildlife. Two piroplasm species have been reported from raccoons (Procyon lotor), Babesia lotori (Babesia sensu stricto clade) and a species related to Babesia microti (called B. microti-like sp.). The goal of this study was to investigate prevalence, distribution, and diversity of Babesia in raccoons. We tested raccoons from selected regions in the United States and Canada for the presence of Babesia sensu stricto and Babesia microti-like sp. piroplasms. Infections of Babesia microti-like sp. were found in nearly all locations sampled, often with high prevalence, while Babesia sensu stricto infections had higher prevalence in the Southeastern United States (20–45% prevalence). Co-infections with both Babesia sp. were common. Sequencing of the partial 18S rRNA and cytochrome oxidase subunit 1 (cox1) genes led to the discovery of two new Babesia species, both found in several locations in the eastern and western United States. One novel Babesia sensu stricto sp. was most similar to Babesia gibsoni while the other Babesia species was present in the ‘western piroplasm’ group and was related to Babesia conradae. Phylogenetic analysis of the cox1 sequences indicated possible eastern and western genetic variants for the three Babesia sensu stricto species. Additional analyses are needed to characterize these novel species; however, this study indicates there are now at least four species of piroplasms infecting raccoons in the United States and Canada (Babesia microti-like sp., Babesia lotori, a novel Babesia sensu stricto sp., a novel western Babesia sp.) and a possible fifth species (Babesia sensu stricto) in raccoons in Japan.

The role of healthy dog carriers of Babesia microti-like piroplasms

BackgroundWhile in Europe Babesia canis has been traditionally held responsible for canine piroplasmosis, Babesia microti-like piroplasm (Bml) infection is being ever more observed in dogs, with the first clinical cases reported in northwestern Spain. This study examines the epidemiological role of healthy dogs living in endemic areas of Bml infection in Spain. The data obtained were used to describe the clinical status and map the geographical distribution of Bml infection in healthy dogs in northwestern Spain.ResultsBlood samples and ticks were taken from 756 healthy dogs representatively across the whole Galicia region (northwestern Spain): stray (n = 211), hunting dogs (n = 333) and pets (n = 212). Blood samples were tested by microscopy parasite observation, nested PCR-RFLP and sequencing. Piroplasm infection prevalences in healthy dogs from northwestern Spain were 17.1% (129/756) by PCR and 3.4% (26/756) by microscopy observation. The species found by PCR were: 2.2% (17/756) for B. canis and 15.1% (114/756) for Bml. Co-infection with B. canis and Bml was noted in 2 dogs. The higher prevalences detected were Bml in hunting dogs (25.5%; 85/333) and B. canis in stray dogs (6.6%; 14/211). In fox-hunting dogs from any area and dogs from the A Coruña Province, significantly higher prevalences of Bml infection were detected (P < 0.001). Upon physical examination, tick infestation was observed: 130 ticks in 18 hunting and three pet dogs. These were subsequently identified as Rhipicephalus sanguineus (s.l.) (49.2%), Ixodes hexagonus (38.5%), Ixodes ricinus (6.9%) and Dermacentor reticulatus (5.4%). Among the more prevalent ticks infesting healthy carrier dogs were I. hexagonus, followed by D. reticulatus and I. ricinus.ConclusionsBabesia canis and Bml were the only piroplasm species found infecting healthy dogs in Galicia, the prevalence of Bml being higher than of B. canis. Factors correlated with a higher Bml infection risk were being a hunting dog and living in the A Coruña Province. Healthy dogs travelling to other countries could act as carriers and probably contribute to the spread of Bml infection in dogs and wild carnivores throughout Europe.

Genetic diversity and prevalence of piroplasm species in equids from Turkey.

Equine piroplasmosis (EP) is a protozoon disease caused by Babesia caballi and Theileria equi transmitted by ticks from the Ixodidae family. This study investigated the genetic heterogeneity and diversity of piroplasm genotypes using the Reverse Line Blotting (RLB) technique for piroplasm species in equids in Turkey. A total of 233 blood samples from 142 horses and 91 donkeys were collected in Şanlıurfa, Tunceli, and Iğdır. The RLB assay was performed for simultaneous detection of piroplasm species. The prevalence of piroplasm infection was 33.5% (95% CI; 27.4-39.9). T. equi was the most common species (%20.2; 95% CI 15.2-25.9), whereas B. caballi infection was not detected. Nine samples hybridized to the Theileria/Babesia and Theileria catch-all probes but did not hybridize to any species-specific probe tested, suggesting the presence of unrecognized Theileria variants or genotypes. Nine samples that gave Theileria spp. signal and two samples that gave T. equi specific signal were sent for sequence analysis and the A and D genotypes of T. equi were found. In addition, T. annulata, B. ovis, and B. canis species were detected in the samples and verified by sequence analysis in Turkey. More research is needed to understand the clinical impact of atypical piroplasm infections in equids.