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Abstract Background SET domain-containing histone lysine methyltransferases (HKMTs) and JmjC domain-containing histone demethylases (JHDMs) are essential for maintaining dynamic changes in histone methylation across parasite development and infection. However, information on the HKMTs and JHDMs in human pathogenic piroplasms, such as Babesia duncani and Babesia microti, and in veterinary important pathogens, including Babesia bigemina, Babesia bovis, Theileria annulata and Theileria parva, is limited. Results A total of 38 putative KMTs and eight JHDMs were identified using a comparative genomics approach. Phylogenetic analysis revealed that the putative KMTs can be divided into eight subgroups, while the JHDMs belong to the JARID subfamily, except for BdJmjC1 (BdWA1_000016) and TpJmjC1 (Tp Muguga_02g00471) which cluster with JmjC domain only subfamily members. The motifs of SET and JmjC domains are highly conserved among piroplasm species. Interspecies collinearity analysis provided insight into the evolutionary duplication events of some SET domain and JmjC domain gene families. Moreover, relative gene expression analysis by RT‒qPCR demonstrated that the putative KMT and JHDM gene families were differentially expressed in different intraerythrocytic developmental stages of B. duncani, suggesting their role in Apicomplexa parasite development. Conclusions Our study provides a theoretical foundation and guidance for understanding the basic characteristics of several important piroplasm KMT and JHDM families and their biological roles in parasite differentiation.

Background: SET domain-containing histone lysine methyltransferases (HKMTs) and JmjC domain-containing histone demethylases (JHDMs) are essential for maintaining dynamic changes in histone methylation across parasite development and infection. However, information on the HKMTs and JHDMs in human pathogenic piroplasms, such as Babesia duncani and Babesia microti, and in veterinary important pathogens, including Babesia bigemina, Babesia bovis, Theileria annulata and Theileria parva, is limited. Results: A total of 38 putative KMTs and eight JHDMs were identified using a comparative genomics approach. Phylogenetic analysis revealed that the putative KMTs can be divided into eight subgroups, while the JHDMs belong to the JARID subfamily, except for BdJmjC1 (BdWA1_000016) and TpJmjC1 (Tp Muguga_02g00471) which cluster with JmjC domain only subfamily members. The motifs of SET and JmjC domains are highly conserved among piroplasm species. Interspecies collinearity analysis provided insight into the evolutionary duplication events of some SET domain and JmjC domain gene families. Moreover, relative gene expression analysis by RT‒qPCR demonstrated that the putative KMT and JHDM gene families were differentially expressed in different intraerythrocytic developmental stages of B. duncani, suggesting their role in Apicomplexa parasite development. Conclusions: Our study provides a theoretical foundation and guidance for understanding the basic characteristics of several important piroplasm KMT and JHDM families and their biological roles in parasite differentiation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

ABSTRACT: Equine piroplasmosis is an apicomplexan hemoprotozoan-caused disease that affects equids worldwide. Horses that survive piroplasmosis can become asymptomatic carriers for the rest of their lives. The present study aimed to determine the frequency of piroplasms and to compare three different blood smear tests (jugular, peripheral, and splenic blood) to detect piroplasms in asymptomatic wandering horses seized by the “Secretaria de Estado da Agricultura, Abastecimento e Desenvolvimento Rural do Distrito Federal” (SEAGRI-DF), Midwest region of Brazil. Of the 100 horses evaluated, 38 were diagnosed positive for piroplasm (38%), with the etiological agents found at the jugular blood smear in 11% (11/100) of horses, peripheral blood smear in 13% (13/100), and splenic blood smear in 38% (38/100). Piroplasm-positive horses showed anemia, neutrophilia, and lymphopenia, but the hematological changes did not differ statistically between positive and negative horses. In summary, the stray horses evaluated showed a high incidence of piroplasm (38%). All positive horses presented one of the etiologic agents in the splenic blood, but some did not show the parasite in the jugular blood smear (27/38, 71%) or the peripheral blood smear (25/38, 65.8%). Thus, the splenic blood was shown to be the best sample to determine the presence of piroplasm in wandering horses. As it is a low-cost and easy-to-perform test, it can be included in the routine diagnosis of equine piroplasmosis, helping to monitor the prevalence of piroplasms in places where molecular techniques are not accessible.

Introduction: Blood parasites pose a significant threat to livestock production in southern Egypt, yet there is a scarcity of information regarding their circulation and epidemiology in sheep in this region. This study aimed to investigate the seroprevalence of blood parasite infections in sheep in Assiut governorate, Upper Egypt. Methods: A total of 400 blood samples were collected from sheep of varying ages and genders. The preliminary screening for the presence of piroplasms, mainly Babesia and Theileria spp., via microscopic examination, followed by investigation of the potential risk factors linked with the exposure to infection. Moreover, molecular identification of both parasites on some of positive samples was performed using PCR targeting Babesia 18S rRNA and Theileria annulata Tams1 gene. Results: The microscopic examination revealed that among the examined sheep, there was an overall prevalence of blood parasites at 44% (176 out of 400), with Babesia spp. observed in 14% (56 out of 400) and Theileria spp. in 30% (120 out of 400). Furthermore, the infection rate was non-significantly higher in young animals (50%) compared to adults (38.5%) (P = 0.246). Male sheep exhibited a significantly higher vulnerability to both parasites’ infection (63.3%) compared to females (35.7%) (P = 0.011). Interestingly, the prevalence of both blood parasites was significantly higher during the cold season (66.1%) compared to the hot season (15.9%) (P = <0.001). The molecular analysis identified the presence of Babesia ovis and Theileria annulata among a subsample of the positive sheep’s bloods films. The identified species were recorded in the GenBankTM databases and assigned specific accession numbers (OQ360720 and OQ360719 for B. ovis), and (OP991838 for T. annulata). Conclusions: Taken together, this study confirms a high prevalence of piroplasmosis and oers epidemiological and molecular insights into blood parasites in sheep from Upper Egypt, highlighting the importance of detecting these parasites in various hosts and their competent vectors (ticks) [ABSTRACT FROM AUTHOR]

Previously, a high prevalence of piroplasms has been reported from Florida pumas (Puma concolor coryi) from southern Florida. In the current study, we describe the biological characteristics of a novel Babesia species in Florida pumas. Ring-stage trophozoites were morphologically similar to trophozoites of numerous small babesids of felids including B. leo, B. felis, and Cytauxzoon felis. Parasitemias in Florida pumas were very low (

IntroductionBlood parasites pose a significant threat to livestock production in southern Egypt, yet there is a scarcity of information regarding their circulation and epidemiology in sheep in this region. This study aimed to investigate the seroprevalence of blood parasite infections in sheep in Assiut governorate, Upper Egypt.MethodsA total of 400 blood samples were collected from sheep of varying ages and genders. The preliminary screening for the presence of piroplasms, mainly Babesia and Theileria spp., via microscopic examination, followed by investigation of the potential risk factors linked with the exposure to infection. Moreover, molecular identification of both parasites on some of positive samples was performed using PCR targeting Babesia 18S rRNA and Theileria annulata Tams1 gene.ResultsThe microscopic examination revealed that among the examined sheep, there was an overall prevalence of blood parasites at 44% (176 out of 400), with Babesia spp. observed in 14% (56 out of 400) and Theileria spp. in 30% (120 out of 400). Furthermore, the infection rate was non-significantly higher in young animals (50%) compared to adults (38.5%) (P = 0.246). Male sheep exhibited a significantly higher vulnerability to both parasites' infection (63.3%) compared to females (35.7%) (P = 0.011). Interestingly, the prevalence of both blood parasites was significantly higher during the cold season (66.1%) compared to the hot season (15.9%) (P = < 0.001). The molecular analysis identified the presence of Babesia ovis and Theileria annulata among a subsample of the positive sheep's bloods films. The identified species were recorded in the GenBank™ databases and assigned specific accession numbers (OQ360720 and OQ360719 for B. ovis), and (OP991838 for T. annulata).ConclusionsTaken together, this study confirms a high prevalence of piroplasmosis and offers epidemiological and molecular insights into blood parasites in sheep from Upper Egypt, highlighting the importance of detecting these parasites in various hosts and their competent vectors (ticks).

Among vector-borne protozoa Hepatozoon felis and Cytauxzoon europaeus are considered emerging species in felids in Europe. To investigate the presence of these two protozoa 127 domestic cats and 4 wildcats were screened by PCRs targeting the 18S rRNA gene of Hepatozoon spp. and piroplasms, as well as the cytb gene of Cytauxzoon spp. The samples were collected inside and outside a region of Hungary, where both protozoan groups are endemic in wildcats. Among domestic cats, one proved to be infected with H. felis. Furthermore, spleen samples of four wildcats were also examined, among which three tested positive for H. felis, and one had co-infection with C. europaeus. Importantly, H. felis from the co-infected wildcat belonged to genogroup II, similarly to H. felis from the positive domestic cat. Based on phylogenetic evidence, this genogroup probably represents a separate species from genogroup I of H. felis, which was hitherto reported from Mediterranean countries in Europe. The two other wildcats also harbored H. felis from genogroup I. Neither Hepatozoon nor Cytauxzoon infections were detected outside the recently discovered endemic region. In conclusion, this study demonstrates for the first time in Europe that H. felis from genogroup II may emerge in free-roaming domestic cats in regions where this protozoan parasite is endemic in wildcats. [ABSTRACT FROM AUTHOR]

We report a case of Babesia microti infection in an immunocompetent child

Background and Purpose: The imidazo[1,2-a] pyridines have huge applications in medicinal chemistry with potent activity against wide spectrum of infectious agents. The efficacy of imidazo[1,2-a]pyridine on the in vitro growth of different piroplasms, including Babesia bovis, B. bigemina, B. divergens, B. caballi, and Theileria equi, was investigated in this study. Methods: The anti-piroplasm efficacy of imidazo[1,2-a] pyridines was assessed using a fluorescence-based SYBR Green I assay. Furthermore, efficacy of imidazo[1,2-a]pyridine against piroplasms following discontinuation of treatment was also assessed using a viability assay. In vitro cultures of B. bovis and T. equi were used to assess the imidazo[1,2-a]pyridine and diminazene aceturate (DA) interaction. Results: In vitro, imidazo[1,2-a]pyridine inhibited the growth of B. bovis, B. bigemina, B. caballi, and T. equi in a dose-dependent manner. The highest inhibitory effects of imidazo[1,2-a]pyridine were detected on the growth of B. caballi with IC50 value of 0.47 ± 0.07. Interestingly, the efficacy of imidazo[1,2-a]pyridine was higher against B. bigemina (IC50: 1.37 ± 0.15) compared to the positive-control DA (IC50: 2.29 ± 0.06). The viability test findings indicate that imidazo[1,2-a]pyridine had a long-lasting inhibitory effect on bovine Babesia parasites in vitro growth up to 4 days after treatment. Notably, when coupled with DA at 0.75 or 0.50 IC50, a high concentration (0.75 IC50) of imidazo[1,2-a]pyridine produced additive suppression of B. bovis growth which suggest that imidazo[1,2-a]pyridine/DA could be a promising combination therapy for the treatment of B. bovis. Conclusion: The obtained encouraging findings pave the way for in vitro and in vivo efficacy trials of imidazo[1,2-a]pyridine derivatives against several piroplasmids. [ABSTRACT FROM AUTHOR]

The American bison (Bison bison) is the largest terrestrial mammal of North America, with around 350,000 individuals in the wild and in private herds but the knowledge regarding the presence of different vector-borne pathigens in these mammals is very poor. Babesia and Theileria spp. are tick-borne apicomplexan parasites which are considered to be among the most commonly found blood parasites of large ruminants, often with a high economic importance. However, the knowledge on piroplasms of bisons is extremely scarce. The aim of our study was to evaluate the presence of apicomplexan parasites in blood and tissues of farmed American bison from Romania. Overall, we tested 222 blood samples and 11 tissues samples (heart, liver, and spleen) from farmed B. bison raised for meat in Romania. All the samples were analyzed by nPCR targeting the 18SrRNA gene for piroplasmids. All positive samples were sequenced and analyzed phylogenetically. The overall prevalence of infection with piroplasmids in American bison was 1.65%, with Babesia divergens and Theileria sp. identified following sequencing. To our knowledge, this is the first report of piroplasms detected in blood and tissues of farmed B. bison from Europe. Further studies are necessary in order to obtain a better overview on the epidemiological status and clinical relevance of piroplasms in farmed American bisons.

Abstract Background Cytauxzoon felis is a life‐threatening protozoan disease of cats. Identification of schizont‐laden macrophages is a point‐of‐care diagnostic test for acute cytauxzoonosis. Hypothesis/Objectives The primary objective determined cytologic agreement between sample types to diagnose acute cytauxzoonosis. The secondary objective evaluated novices' ability to identify cytauxzoon organisms in blood films and tissue aspirates. Animals Thirty‐eight cats with suspected acute cytauxzoonosis and 5 controls examined postmortem. Methods Cases were prospectively submitted and collected. Blood film, lymph node, and splenic aspirates were blindly reviewed for sample quality, presence of schizont‐laden macrophages, and agreement between sample types. A subset of cases and controls were evaluated by 12 blinded novice observers to determine sensitivity and specificity for identifying organisms in various sample types. Results Acute cytauxzoonosis diagnosis was made on at least 1 sample type in 28/38 cats. Schizont‐laden macrophages were seen on 33% (10/30) of blood films, 56% (19/34) lymph node aspirates, 77% (26/34) splenic aspirates. Schizont‐laden macrophages were more likely seen on splenic than lymph node aspirates (McNemar's, P = .03) or blood film (McNemar's, P =

Theileria orientalis is an emerging apicomplexan pathogen of cattle occurring in areas populated by the principal vector tick, Haemaphysalis longicornis. Unlike transforming Theileria spp. that induce cancer-like proliferation of lymphocytes via their schizont stage, T. orientalis destroys host erythrocytes during its piroplasm phase resulting in anaemia. The underlying pathogenic processes of T. orientalis infection are poorly understood; consequently, there are no vaccines for prevention of T. orientalis infection and chemotherapeutic options are limited. To identify antigens expressed during the piroplasm phase of T. orientalis, including those which may be useful targets for future therapeutic development, we examined the proteome across three common genotypes of the parasite (Ikeda, Chitose and Buffeli) using preparations of piroplasms purified from bovine blood. A combination of Triton X-114 extraction, one-dimensional electrophoresis and LC-MS/MS identified a total of 1113 proteins across all genotypes, with less than 3% of these representing host-derived proteins. Just over three quarters of T. orientalis proteins (78%) identified were from the aqueous phase of the TX-114 extraction representing cytosolic proteins, with the remaining 22% from the detergent phase, representing membrane-associated proteins. All enzymes involved in glycolysis were expressed, suggesting that this is the major metabolic pathway used during the T. orientalis piroplasm phase. Proteins involved in binding and breakdown of haemoglobin were also identified, suggesting that T. orientalis uses haemoglobin as a source of amino acids. A number of proteins involved in host cell interaction were also identified which may be suitable targets for the development of chemotherapeutics or vaccines. [ABSTRACT FROM AUTHOR]

Abstract Background Babesia spp. are protozoan parasites of great medical and veterinary importance, especially in the northern Hemisphere. Ticks are known vectors of Babesia spp., although some Babesia-tick interactions have not been fully elucidated. Methods The present review was performed to investigate the specificity of Babesia-tick species interactions that have been identified using molecular techniques in studies conducted in the last 20 years under field conditions. We aimed to indicate the main vectors of important Babesia species based on published research papers (n = 129) and molecular data derived from the GenBank database. Results Repeated observations of certain Babesia species in specific species and genera of ticks in numerous independent studies, carried out in different areas and years, have been considered epidemiological evidence of established Babesia-tick interactions. The best studied species of ticks are Ixodes ricinus, Dermacentor reticulatus and Ixodes scapularis (103 reports, i.e. 80% of total reports). Eco-epidemiological studies have confirmed a specific relationship between Babesia microti and Ixodes ricinus, Ixodes persulcatus, and Ixodes scapularis and also between Babesia canis and D. reticulatus. Additionally, four Babesia species (and one genotype), which have different deer species as reservoir hosts, displayed specificity to the I. ricinus complex. Eco-epidemiological studies do not support interactions between a high number of Babesia spp. and I. ricinus or D. reticulatus. Interestingly, pioneering studies on other species and genera of ticks have revealed the existence of likely new Babesia species, which need more scientific attention. Finally, we discuss the detection of Babesia spp. in feeding ticks and critically evaluate the data on the role of the latter as vectors. Conclusions Epidemiological data have confirmed the specificity of certain Babesia-tick vector interactions. The massive amount of data that has been thus far collected for the most common tick species needs to be complemented by more intensive studies on Babesia infections in underrepresented tick species. Graphical abstract

Previously, a high prevalence of piroplasms has been reported from Florida pumas (Puma concolor coryi) from southern Florida. In the current study, we describe the biological characteristics of a novel Babesia species in Florida pumas. Ring-stage trophozoites were morphologically similar to trophozoites of numerous small babesids of felids including B. leo, B. felis , and Cytauxzoon felis. Parasitemias in Florida pumas were very low (<1%) and hematologic values of 25 Babesia- infected Florida pumas were within normal ranges for P. concolor. Phylogenetic analysis of near full-length 18S rRNA gene, β-tubulin, cytochrome c oxidase subunit I, cytochrome c oxidase subunit III, and cytochrome b gene sequences indicated that this Babesia species is a member of the Babesia sensu stricto clade and is related to groups of Babesia spp. from carnivores or ungulates, although the closest group varied by gene target. Internal transcribed spacer (ITS)-1 region sequences from this Babesia sp. from 19 Florida pumas were 85.7–99.5% similar to each other and ∼88% similar to B. odocoilei. Similarly, an ITS-2 sequence from one puma was 96% similar to B. bigemina and 92% similar to a Babesia sp. from a red panda (Ailurus fulgens). Infected pumas were positive for antibodies that reacted with B. odocoilei , B. canis, and B. bovis antigens with titers of 1:256, 1:128, and 1:128, respectively. No serologic reactivity was noted for Theileria equi. No molecular evidence of congenital infection was detected in 24 kittens born to 11 Babesia -infected female pumas. Pumas from other populations in the United States [Louisiana (n = 1), North Dakota (n = 5) and Texas (n = 28)], British Columbia, Canada (n = 9), and Costa Rica (n = 2) were negative for this Babesia sp. Collectively, these data provide morphologic, serologic, genetic, and natural history data for this novel Babesia sp. which we propose the name Babesia coryicola sp. nov. sp. This is the first description of a felid-associated Babesia species in North America. [Display omitted] • Novel species of small Babesia described from the Florida puma. • Babesia coryicola sp. nov. was phylogenetically included in the Babesia sensu stricto. • Prevalence of Babesia coryicola sp. nov. in the Florida puma was high. • No clinical abnormalities noted in infected Florida pumas. [ABSTRACT FROM AUTHOR]

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 < 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.

Among vector-borne protozoa Hepatozoon felis and Cytauxzoon europaeus are considered emerging species in felids in Europe. To investigate the presence of these two protozoa 127 domestic cats and 4 wildcats were screened by PCRs targeting the 18S rRNA gene of Hepatozoon spp. and piroplasms, as well as the cytb gene of Cytauxzoon spp. The samples were collected inside and outside a region of Hungary, where both protozoan groups are endemic in wildcats. Among domestic cats, one proved to be infected with H. felis. Furthermore, spleen samples of four wildcats were also examined, among which three tested positive for H. felis, and one had co-infection with C. europaeus. Importantly, H. felis from the co-infected wildcat belonged to genogroup II, similarly to H. felis from the positive domestic cat. Based on phylogenetic evidence, this genogroup probably represents a separate species from genogroup I of H. felis, which was hitherto reported from Mediterranean countries in Europe. The two other wildcats also harbored H. felis from genogroup I. Neither Hepatozoon nor Cytauxzoon infections were detected outside the recently discovered endemic region. In conclusion, this study demonstrates for the first time in Europe that H. felis from genogroup II may emerge in free-roaming domestic cats in regions where this protozoan parasite is endemic in wildcats.

Background: Cytauxzoon felis is a life‐threatening protozoan disease of cats. Identification of schizont‐laden macrophages is a point‐of‐care diagnostic test for acute cytauxzoonosis. Hypothesis/Objectives: The primary objective determined cytologic agreement between sample types to diagnose acute cytauxzoonosis. The secondary objective evaluated novices' ability to identify cytauxzoon organisms in blood films and tissue aspirates. Animals Thirty‐eight cats with suspected acute cytauxzoonosis and 5 controls examined postmortem. Methods: Cases were prospectively submitted and collected. Blood film, lymph node, and splenic aspirates were blindly reviewed for sample quality, presence of schizont‐laden macrophages, and agreement between sample types. A subset of cases and controls were evaluated by 12 blinded novice observers to determine sensitivity and specificity for identifying organisms in various sample types. Results: Acute cytauxzoonosis diagnosis was made on at least 1 sample type in 28/38 cats. Schizont‐laden macrophages were seen on 33% (10/30) of blood films, 56% (19/34) lymph node aspirates, 77% (26/34) splenic aspirates. Schizont‐laden macrophages were more likely seen on splenic than lymph node aspirates (McNemar's, P =.03) or blood film (McNemar's, P = <.001). Novice observers were more likely to agree with experts when identifying schizont‐laden macrophages in splenic aspirates (sensitivity = 77.1%, specificity = 94.4%) versus lymph node aspirates (sensitivity = 52.8%, specificity = 96.4%) or blood films (sensitivity = 41.7%, specificity = 96.9%). Conclusions and Clinical Importance: Schizont‐laden macrophages are most frequently identified in spleen, even by novice observers. If the diagnosis of acute cytauxzoonosis cannot be confirmed via blood film, then splenic, followed by peripheral lymph node aspirates can be considered. [ABSTRACT FROM AUTHOR]

BackgroundEmerging long reads sequencing technology has greatly changed the landscape of whole-genome sequencing, enabling scientists to contribute to decoding the genetic information of non-model species. The sequences generated by PacBio or Oxford Nanopore Technology (ONT) be assembled de novo before further analyses. Some genome de novo assemblers have been developed to assemble long reads generated by ONT. The performance of these assemblers has not been completely investigated. However, genome assembly is still a challenging task.Methods and ResultsWe systematically evaluated the performance of nine de novo assemblers for ONT on different coverage depth datasets. Several metrics were measured to determine the performance of these tools, including N50 length, sequence coverage, runtime, easy operation, accuracy of genome and genomic completeness in varying depths of coverage. Based on the results of our assessments, the performances of these tools are summarized as follows: 1) Coverage depth has a significant effect on genome quality; 2) The level of contiguity of the assembled genome varies dramatically among different de novo tools; 3) The correctness of an assembled genome is closely related to the completeness of the genome. More than 30× nanopore data can be assembled into a relatively complete genome, the quality of which is highly dependent on the polishing using next generation sequencing data.ConclusionConsidering the results of our investigation, the advantage and disadvantage of each tool are summarized and guidelines of selecting assembly tools are provided under specific conditions.

Babesiosis developed in a 62-year-old immunocompetent physician, who had an uneventful recovery after receiving atovaquone and azithromycin. Three years later, babesiosis developed again, and he was again successfully given treatment. Clinical and laboratory evidence were highly supportive of Babesia reinfection. Healthcare professionals should be aware that reinfection might occur in babesiosis. [ABSTRACT FROM AUTHOR]

Background: Babesia spp. are protozoan parasites of great medical and veterinary importance, especially in the northern Hemisphere. Ticks are known vectors of Babesia spp., although some Babesia-tick interactions have not been fully elucidated. Methods: The present review was performed to investigate the specificity of Babesia-tick species interactions that have been identified using molecular techniques in studies conducted in the last 20 years under field conditions. We aimed to indicate the main vectors of important Babesia species based on published research papers (n = 129) and molecular data derived from the GenBank database. Results: Repeated observations of certain Babesia species in specific species and genera of ticks in numerous independent studies, carried out in different areas and years, have been considered epidemiological evidence of established Babesia-tick interactions. The best studied species of ticks are Ixodes ricinus, Dermacentor reticulatus and Ixodes scapularis (103 reports, i.e. 80% of total reports). Eco-epidemiological studies have confirmed a specific relationship between Babesia microti and Ixodes ricinus, Ixodes persulcatus, and Ixodes scapularis and also between Babesia canis and D. reticulatus. Additionally, four Babesia species (and one genotype), which have different deer species as reservoir hosts, displayed specificity to the I. ricinus complex. Eco-epidemiological studies do not support interactions between a high number of Babesia spp. and I. ricinus or D. reticulatus. Interestingly, pioneering studies on other species and genera of ticks have revealed the existence of likely new Babesia species, which need more scientific attention. Finally, we discuss the detection of Babesia spp. in feeding ticks and critically evaluate the data on the role of the latter as vectors. Conclusions: Epidemiological data have confirmed the specificity of certain Babesia-tick vector interactions. The massive amount of data that has been thus far collected for the most common tick species needs to be complemented by more intensive studies on Babesia infections in underrepresented tick species. [ABSTRACT FROM AUTHOR]

The protozoan parasites Theileria equi and Babesia caballi, transmitted by ticks, cause equine piroplasmosis, the most prevalent tick-borne disease in equids. Trichinellosis is a worldwide food-borne zoonosis caused by helminth Trichinella spp. that can lead to serious disease in humans, with fatal outcome. Although the infection is rare in horses, it deserves attention due to the increasing use of horse meat as a source of protein for humans. Horse trichinellosis is caused by several Trichinella species, most commonly by T. spiralis. The aim of the study was to determine the prevalence of antibodies to T. equi, B. caballi and Trichinella spp. in equids from three states of Northern Nigeria. Serum samples were collected from 139 clinically healthy animals, comprising 115 horses and 24 donkeys. Antibodies to T. equi and B. caballi were detected in serum by competitive-inhibition enzyme-linked immunosorbent assay (cELISA) and antibodies to Trichinella spp. by ELISA. Antibodies to T. equi were detected in 34% of equids (41% horses and 0% donkeys), antibodies to B. caballi in 9% of equids (8% horses and 13% donkeys), and antibodies to Trichinella spp. in 4% of equids (4% horses and 0% donkeys). There was co-infection of T. equi and B. caballi in 1% of horses and co-infection of T. equi and Trichinella spp. in 2.6% of horses. This is the first report on seroprevalence of Trichinella spp. in equids from Northern Nigeria. [ABSTRACT FROM AUTHOR]

Background: Emerging long reads sequencing technology has greatly changed the landscape of whole-genome sequencing, enabling scientists to contribute to decoding the genetic information of non-model species. The sequences generated by PacBio or Oxford Nanopore Technology (ONT) be assembled de novo before further analyses. Some genome de novo assemblers have been developed to assemble long reads generated by ONT. The performance of these assemblers has not been completely investigated. However, genome assembly is still a challenging task. Methods and Results: We systematically evaluated the performance of nine de novo assemblers for ONT on different coverage depth datasets. Several metrics were measured to determine the performance of these tools, including N50 length, sequence coverage, runtime, easy operation, accuracy of genome and genomic completeness in varying depths of coverage. Based on the results of our assessments, the performances of these tools are summarized as follows: 1) Coverage depth has a significant effect on genome quality; 2) The level of contiguity of the assembled genome varies dramatically among different de novo tools; 3) The correctness of an assembled genome is closely related to the completeness of the genome. More than 30× nanopore data can be assembled into a relatively complete genome, the quality of which is highly dependent on the polishing using next generation sequencing data. Conclusion: Considering the results of our investigation, the advantage and disadvantage of each tool are summarized and guidelines of selecting assembly tools are provided under specific conditions. [ABSTRACT FROM AUTHOR]

Abstract Background Understanding how fauna translocation and antiparasitic drug treatment impact parasite community structure within a host is vital for optimising translocation outcomes. Trypanosoma spp. and piroplasms (Babesia and Theileria spp.) are known to infect Australian marsupials, including the woylie (Bettongia penicillata). However relatively little is known about these haemoparasites, or how they respond to management practices such as translocation. We monitored haemoparasites infecting woylies for up to 12 months during two fauna translocations to supplement existing woylie populations in three different sites (Dryandra, Walcott and Warrup East) within south-western Australia between 2014 and 2016, with the aim of investigating (i) how haemoparasite prevalence, Trypanosoma spp. richness and Trypanosoma spp. community composition varied over time and between different sites following translocation; and (ii) whether ivermectin treatment indirectly impacts haemoparasite prevalence. Using molecular methods, 1211 blood samples were screened for the presence of trypanosomes, and a subset of these samples (n = 264) were also tested for piroplasms. Results Trypanosomes and piroplasms were identified in 55% and 94% of blood samples, respectively. We identified five Trypanosoma species, two Theileria species, a single species of Babesia and a novel Bodo species. Trypanosoma spp. richness and the prevalence of haemoparasite co-infection increased after translocation. Prior to translocation, Trypanosoma spp. community composition differed significantly between translocated and resident woylies within Walcott and Warrup East, but not Dryandra. Six months later, there was a significant difference between translocated and resident woylies within Dryandra, but not Walcott or Warrup East. The response of haemoparasites to translocation was highly site-specific, with predominant changes to the haemoparasite community in translocated woylies occurring within the first few months following translocation. Ivermectin treatment had no significant effect on haemoparasite prevalence. Conclusions This study contributes to our understanding of haemoparasite dynamics in woylies following translocation. The highly site-specific and rapid response of haemoparasites to translocation highlights the need to better understand what drives these effects. Given that haemoparasite prevalence and composition of translocated and resident animals changed significantly following translocation, we propose that parasite monitoring should form an essential component of translocation protocols, and such protocols should endeavour to monitor translocated hosts and cohabiting species.

Abstract Background Vector-borne diseases exert a global economic impact to the livestock industry. Understanding how agriculture practices and acaricide usage affect the ecology of these diseases is important for making informed management decisions. Theileria cervi is a hemoprotozoan parasite infecting white-tailed deer (Odocoileus virginianus) and is transmitted by the lone star tick, Amblyomma americanum. The purpose of this study was to determine if acaricide treatment decreased hematozoan prevalence in farmed white-tailed deer when compared to geographically-close wild deer or altered the genotypes of T. cervi present. Results We compared prevalence of T. cervi in 52 farmed adult white-tailed deer which were regularly treated with permethrin and ivermectin, 53 farmed neonates that did not receive treatment for vector control, and 42 wild deer that received no form of chemical vector control. Wild deer had significantly higher prevalence of T. cervi than farmed deer. Additionally, no neonate fawns tested positive for T. cervi, and we found that age was a significant predictor of infection status. We found no difference in genotypic variation in T. cervi isolates between adjacent herds of farmed and wild white-tailed deer, although a divergent genotype X was identified. Chronic infection with T. cervi had no significant effects on mortality in the white-tailed deer. Conclusions We found significantly lower prevalence of T. cervi infection in farmed (40%) compared to wild white-tailed deer (98%), which may be due to the inclusion of chemical vector control strategies. More work is needed to determine the implications, if any, of mixed genotypic infections of T. cervi, although we found no significant effect of infection with Theileria on mortality in farmed deer. Theileria infection does sometimes cause disease when an animal is stressed, immunosuppressed, or translocated from non-endemic to endemic regions.

Theileria orientalis is an emerging apicomplexan pathogen of cattle occurring in areas populated by the principal vector tick, Haemaphysalis longicornis. Unlike transforming Theileria spp. that induce cancer-like proliferation of lymphocytes via their schizont stage, T. orientalis destroys host erythrocytes during its piroplasm phase resulting in anaemia. The underlying pathogenic processes of T. orientalis infection are poorly understood; consequently, there are no vaccines for prevention of T. orientalis infection and chemotherapeutic options are limited. To identify antigens expressed during the piroplasm phase of T. orientalis, including those which may be useful targets for future therapeutic development, we examined the proteome across three common genotypes of the parasite (Ikeda, Chitose and Buffeli) using preparations of piroplasms purified from bovine blood. A combination of Triton X-114 extraction, one-dimensional electrophoresis and LC-MS/MS identified a total of 1113 proteins across all genotypes, with less than 3% of these representing host-derived proteins. Just over three quarters of T. orientalis proteins (78%) identified were from the aqueous phase of the TX-114 extraction representing cytosolic proteins, with the remaining 22% from the detergent phase, representing membrane-associated proteins. All enzymes involved in glycolysis were expressed, suggesting that this is the major metabolic pathway used during the T. orientalis piroplasm phase. Proteins involved in binding and breakdown of haemoglobin were also identified, suggesting that T. orientalis uses haemoglobin as a source of amino acids. A number of proteins involved in host cell interaction were also identified which may be suitable targets for the development of chemotherapeutics or vaccines.

Theileriosis in bovines leads to economical burden to livestock farmers due to mortality, treatment costs and diminished productivity of affected animals. This manuscript reports about diagnosis of theileriosis in crossbred dairy cattle of Southern Haryana and its successful therapeutic management. Blood samples were received at Disease Investigation Laboratory, Lala Lajpat Rai University of Veterinary and Animal Sciences, Mahendragarh, Haryana from crossbred dairy cattle of Southern Haryana having a clinical history of fever, anorexia, weakness, anaemia and lymphadenopathy. Out of 34 suspected blood samples of cattle received, 11 samples were found positive for theileriosis by the identification of piroplasms in erythrocytes. The positive animals were treated with buparvaquone along with supportive therapy. The affected animals were found as recovered from fever and become normal within a week of treatment. The timely diagnosis and early treatment for haemoprotozoan infections can reduce the mortality rate as well as morbidity rate among the valuable livestock. [ABSTRACT FROM AUTHOR]

Theileria parva is a protozoan parasite that causes East Coast fever (ECF), an economically important disease of cattle in Africa. It is transmitted mainly by the tick Rhipicephalus appendiculatus. Research efforts to develop a subunit vaccine based on parasite neutralizing antibodies and cytotoxic T-lymphocytes have met with limited success. The molecular mechanisms underlying T. parva life cycle stages in the tick vector and bovine host are poorly understood, thus limiting progress toward an effective and efficient control of ECF. Transcriptomics has been used to identify candidate vaccine antigens or markers associated with virulence and disease pathology. Therefore, characterization of gene expression throughout the parasite's life cycle should shed light on host–pathogen interactions in ECF and identify genes underlying differences in parasite stages as well as potential, novel therapeutic targets. Recently, the first gene expression profiling of T. parva was conducted for the sporoblast, sporozoite, and schizont stages. The sporozoite is infective to cattle, whereas the schizont is the major pathogenic form of the parasite. The schizont can differentiate into piroplasm, which is infective to the tick vector. The present study was designed to extend the T. parva gene expression profiling to the piroplasm stage with reference to the schizont. Pairwise comparison revealed that 3,279 of a possible 4,084 protein coding genes were differentially expressed, with 1,623 (49%) genes upregulated and 1,656 (51%) downregulated in the piroplasm relative to the schizont. In addition, over 200 genes were stage-specific. In general, there were more molecular functions, biological processes, subcellular localizations, and pathways significantly enriched in the piroplasm than in the schizont. Using known antigens as benchmarks, we identified several new potential vaccine antigens, including TP04_0076 and TP04_0640, which were highly immunogenic in naturally T. parva-infected cattle. All the candidate vaccine antigens identified have yet to be investigated for their capacity to induce protective immune response against ECF.

This study, conducted in a nature reserve in southern Portugal, investigated the frequency and diversity of tick-borne piroplasms in six species of adult ixodid ticks removed from 71 fallow deer (Dama dama) and 12 red deer (Cervus elaphus), collected over the period 2012–2019. The majority of 520 ticks were Ixodes ricinus (78.5%), followed by Rhipicephalus sanguineus sensu lato, Hyalomma lusitanicum, Haemaphysalis punctata, Dermacentor marginatus, and Ixodes hexagonus. The R. sanguineus ticks collected from the deer were clearly exophilic, in contrast to the endophilic species usually associated with dogs. Four tick-borne piroplasms, including Theileria spp., and the zoonotic species, Babesia divergens and Babesia microti, were detected. B. divergens 18S rDNA, identical to that of the bovine reference strain U16370 and to certain strains from red deer, was detected in I. ricinus ticks removed from fallow deer. The sporadic detection of infections in ticks removed from the same individual hosts suggests that the piroplasms were present in the ticks rather than the hosts. Theileria sp. OT3 was found in I. ricinus and, along with T. capreoli, was also detected in some of the other tick species. The natural vector and pathogenic significance of this piroplasm are unknown.

Over the last two decades, Cytauxzoon spp. has been conquering Eurasia, although this fact has only been brought to light through recent more intensive research after the discovery of C. manul in Pallas’ cat. In Europe, Cytauxzoon was detected mainly in southern countries and later in central Europe. This pathogen has now been found in Russia for the first time (50 km from Moscow), this being the most northern sighting in Eurasia. A captive serval (Leptailurus serval) was found to be infected. Hematological analysis showed a crucial decrease in the number of leukocytes and erythrocytes, as well as in hemoglobin concentration. Genetic analysis confirmed the presence of Cytauxzoon spp. in serval blood at the beginning of the disease period. The identical pathogen was found in one bobcat at the same breeding center. Two other haplotypes of Cytauxzoon spp. were obtained from domestic cats at the same location, identical to the samples from Italy. One new haplotype, which was sequenced for the first time, was found in 7/7 investigated Amur wildcats (100%). The high occurrence and diversity of these pathogens suggest that they are present in free-ranging domestic cats and wild felids in Russia, and may be considered a potential threat to the endangered species. Current research shows that the genetic diversity of this pathogen may be even higher than it was suggested previously. Further genetic research is necessary to describe the diversity and phylogeny of this pathogen in Eurasia.

Abstract Background Cytauxzoonosis is an emerging tick-borne disease of domestic and wild felids. Cytauxzoon felis induces severe and often fatal disease in domestic cats. In Europe, clinical and subclinical infections caused by Cytauxzoon sp. are described. We report the first cases of Cytauxzoon sp. infection in domestic cats in Switzerland. Methods Clinical and laboratory data and results of PCR analyses were collected from Cytauxzoon sp. PCR-positive cats and the cats followed for up to 851 days. Results The cases were three two-month old kittens from the same litter (Cases 1–3) and two adult domestic shorthair cats (Cases 4 and 5). The cats originated from the north-west and west of Switzerland. Cases 1–3 presented with moderate to severe regenerative anaemia and intraerythrocytic inclusions. Cytauxzoon sp. was confirmed by PCR and sequencing. The kittens made a clinical and haematological recovery after blood transfusion and/or treatment with azithromycin and atovaquone, but erythroparasitaemia persisted. Case 4 presented with severe non-regenerative anaemia. Case 5 was healthy and used as a blood donor for Case 4. Following blood transfusion, Case 4 showed intraerythrocytic inclusions, and Cytauxzoon sp. was confirmed in both Cases 4 and 5 using PCR and sequencing. Case 4 achieved clinical and haematological remission after treatment with azithromycin, atovaquone and immunosuppressive drugs. Eight months later, Case 4 was presented again with anaemia but tested Cytauxzoon sp. PCR-negative. Sequencing of 1637 bp of the 18S rRNA gene of Cytauxzoon sp. revealed 100% nucleotide sequence identity among isolates of Cases 1–3 and between isolates of Cases 4 and 5, and 99% sequence identity between isolates of all cases. Phylogenetic analysis revealed the closest relationship of the Swiss isolates to Cytauxzoon sp. isolates from domestic cats and wild felids from France, Spain and Romania and to Cytauxzoon manul from a Pallas’s cat. Conclusions This is the first report of Cytauxzoon sp. infection in domestic cats in Switzerland. It is also the first report of infection in very young kittens and transmission of Cytauxzoon sp. to an adult cat by transfusion of blood from an asymptomatic cat. The cats recovered but some developed chronic asymptomatic erythroparasitaemia for up to 28 months. Domestic cats may act as reservoirs for Cytauxzoon sp. in Europe and blood donor cats should be screened for this agent by PCR.

Cytauxzoon felis, a tick-borne hemoprotozoal pathogen of felids, causes an acute, often-fatal disease in domestic cats. While public awareness of the disease has increased, few studies have evaluated the incidence of acute cytauxzoonosis cases and their associated risk factors. The objective of this study was to retrospectively review records of cats diagnosed with acute cytauxzoonosis in eastern Kansas from 2006-2019 using clinic records and determine: (i) feline cytauxzoonosis risk factors; and (ii) if cytauxzoonosis case incidence is increasing. Although inter-annual variation of acute cytauxzoonosis diagnosis was observed in the eastern Kansas domestic cat population, the overall incidence trend remained largely unchanged over the 14-year case review period. In comparison to ill (C. felis-unrelated) control cases, more acute cytauxzoonosis cases were diagnosed in spring and summer, suggesting a seasonal fluctuation of infection, with samples most commonly submitted from ≥1 year old, owned, male cats. Although cytauxzoonosis case submissions remained consistent over the broad study period, increasing tick vector and domestic cat reservoir populations may contribute to additional cytauxzoonosis case expansion in endemic areas. Investigating the incidence of acute cytauxzoonosis, patient risk factors, and ecological variables that influence disease transmission are important steps towards developing and communicating the need for effective cytauxzoonosis control strategies for high-risk cat populations. [ABSTRACT FROM AUTHOR]

Increasing amount of data attest that (in the context of vector-borne infections) birds are not only important as hosts of blood-sucking arthropod vectors, but also as reservoirs of vector-borne pathogens. From 2015 to 2019 cadavers of 100 birds (from 45 species, nine orders) were collected in Hungary, and their organs were screened for DNA from a broad range of vector-borne bacteria with PCR and sequencing. Molecular analyses revealed the presence of Anaplasmataceae, and sequencing identified bacteria closely related to Neorickettsia helminthoeca and Ehrlichia chaffeensis in a Eurasian teal (Anas crecca) and a song thrush (Turdus philomelos), respectively. All samples were PCR negative for rickettsiae, borreliae, Francisella and Coxiella spp., as well as for piroplasms. To our knowledge, this is the first report of a Neorickettsia and an Ehrlichia sp., which belong to the phylogenetic groups of N. helminthoeca and E. chaffeensis, respectively, from Europe. The potential presence of these two vector-borne bacteria needs to be taken into account during future studies on the eco-epidemiology of Anaplasmataceae in Europe. [ABSTRACT FROM AUTHOR]

The archived blood of rodents species such as Meriones, Mus, and Microtus species was investigated in order to detect any piroplasms species. In this study 18S rRNA target gene of piroplasm parasite was amplified by PCR in a Meriones persicus; so, the Locus 1 and Locus 2 of 18S rRNA were sequenced, successfully. A Theileria-like taxa was suspected in accordance with the BLAST analysis of 18S rRNA L1 and L2 with 96% and 91% sequence homology, respectively. The present study was the first report of a Theileria-like species in M. persicus from Iran. [ABSTRACT FROM AUTHOR]

Human babesiosis is a life-threatening infectious disease that causes societal and economic impact worldwide. Several species of Babesia cause babesiosis in terrestrial vertebrates, including humans. A one-day clinic was held in Ontario, Canada, to see if a red blood cell parasite, which is present in blacklegged ticks, Ixodes scapularis, is present in humans. Based on PCR testing and DNA sequencing of the 18S rRNA gene, we unveiled B. odocoilei in two of 19 participants. DNA amplicons from these two patients are almost identical matches with the type strains of B. odocoilei in GenBank. In addition, the same two human subjects had the hallmark symptoms of human babesiosis, including night sweats, chills, fevers, and profound fatigue. Based on symptoms and molecular identification, we provide substantive evidence that B. odocoilei is pathogenic to humans. Dataset reveals that B. odocoilei serologically cross-reacts with Babesia duncani. Clinicians must realize that there are more than two Babesia spp. in North America that cause human babesiosis. This discovery signifies the first report of B. odocoilei causing human babesiosis.

The published literature on schizont-"transforming," or pathogenic theileriosis, in African wild artiodactyls is dated and based on limited information. Here the authors review the taxonomy, diagnosis, epidemiology, hematology, pathology, and aspects of control in various species. Molecular studies based on 18S and 16S rRNA gene sequences have shown that African wild artiodactyls are commonly infected with diverse Theileria spp., as well as nontheilerial hemoprotozoa and rickettsia-like bacteria, and coinfections with pathogenic and nonpathogenic Theileria species are often recorded. Although theileriosis is still confusingly referred to as cytauxzoonosis in many species, the validity of a separate Cytauxzoon genus in artiodactyls is debated. The epidemiology of theileriosis is complex; the likelihood of fatal disease depends on the interplay of parasite, vertebrate host, tick vector, and environmental factors. Roan calves (Hippotragus equinus) and stressed animals of all host species are more susceptible to fatal theileriosis. Even though regenerative anemia is common, peripheral blood piroplasm parasitemia does not correlate with disease severity. Other than anemia, common macroscopic lesions include icterus, hemorrhages (mucosal, serosal, and tissue), fluid effusions into body cavities, lung edema, and variably sized raised cream-colored foci of leukocyte infiltration in multiple organs. Histopathologic findings include vasocentric hyperproliferation and lysis of atypical leukocytes with associated intracellular schizonts, parenchymal necrosis, hemorrhage, thromboembolism, and edema. Immunophenotyping is required to establish the identity of the schizont-transformed leukocytes in wild ungulates. Throughout the review, we propose avenues for future research by comparing existing knowledge on selected aspects of theileriosis in domestic livestock with that in African wild artiodactyls. [ABSTRACT FROM AUTHOR]

Translocation of animals in fragmented habitats is an important means of dispersal and gene flow, however, the movement of animals has led to the spread of various diseases globally and wildlife are often the reservoirs of these diseases. Currently, Cape mountain zebra are translocated within South Africa as a management method for augmentation of isolated and fragmented populations. The movement of pathogens due to translocations in local regions have gone largely unchecked, particularly where there may still be isolated regions that can be negatively affected. Equine piroplasmosis is a tick-borne disease caused by Theilaria equi and/or Babesia caballi reported to occur in equids (Bhoora et al., 2010; Zweygarth et al., 2002). Here, the presence of T. equi and B. caballi was detected in 137 clinically healthy Cape mountain zebra from three South African reserves, Mountain Zebra National Park (MZNP), De Hoop Nature Reserve (DHNR) and Karoo National Park (KNP) using the multiplex EP real-time PCR (qPCR) assay. We observed 100% prevalence for T. equi and identified only one animal from MZNP with B. caballi. These results affirm that precautions should be taken prior to founding new populations of Cape mountain zebra and that potential farms and properties adjacent to prospective reserves should be screened for the presence of the organisms in order to mitigate risks of infection to domestic animals. Image 1 • Piroplasms are prevalent in 3 Nature reserves containing Cape mountain zebra. • Translocation of Cape mountain zebra leads to risk of spread of equine piroplasms. • Equids in adjacent areas to new populations of Cape mountain zebra are at risk of infection. [ABSTRACT FROM AUTHOR]

Protozoan parasites of the genus Babesia and Theileria are significant tick-borne pathogens of domestic animals and cause economic losses to the livestock industry in tropical and subtropical regions worldwide. In this study, 274 blood samples and 32 tick samples were collected from four counties of Wuwei City in northwestern China in June and July in 2018. The DNA from the field samples was analyzed for Babesia or Theileria infection using specific PCR and sequencing based on 18S rRNA gene fragments. The total infection rates were 0.4% for B. motasi and T. separata (both 1/274) in sheep, 3.1% for T. annulata (1/32), 6.2% for B. occultans (2/32) and 9.4% for B. bigemina (3/32) in ticks, respectively. In particular, T. separata has been for the first time detected in sheep in China and B. occultans in Hyalomma asiaticum from Gansu Province of China. [ABSTRACT FROM AUTHOR]

Abstract Rangeliosis, caused by protozoan Rangelia vitalii , is transmitted by the tick Amblyomma aureolatum. The disease is characterized by hemolytic and hemorrhagic disorder and has been described in dogs and other wild canids. The aim of this study was to compare clinicopathological findings and laboratory results of a Rangelia infection in a crab-eating fox (Cerdocyon thous) with those of canine rangeliosis. The zoo of Universidade de Caxias do Sul, received a crab-eating fox with marked jaundice in mucous membranes, dark-colored stools and neurological signs. The animal underwent an ear tip smear examination and blood collection for complete blood counts, serum biochemistry and PCR. Free-living and intraerythrocytic pyriform structures consistent with R. vitalii were found in the blood smear of the ear tip. The erythrogram revealed normocytic normochromic anemia, moderate macrocytosis, polychromasia and metarubricytosis. The leukogram revealed leukocytosis with neutrophilia and monocytosis, as well as severe thrombocytopenia. Serum biochemistry showed hypoproteinemia, hypoalbuminemia and elevated levels of urea and creatinine. The treatment was performed with imidocarb hydrochloride and dexamethasone, however 24 h after initiation of treatment the animal died. Macroscopic examination revealed jaundice, subcutaneous edema, enlarged superficial lymph nodes, splenomegaly, and hemorrhage of internal organs. Histological sections of the cerebellum, lung, pancreas, intestine and heart were consistent with R. vitalii infection of the vascular endothelium. Pathological and hematological findings were similar to those found in infected dogs, with clinical presentation characterized by hemolytic anemia and hemorrhage. The description of this case showed that C. thous does not only serve as reservoir of R. vitalii but may also develop disease. [ABSTRACT FROM AUTHOR]

Tick-borne zoonotic diseases have an economic and societal impact on the well-being of people worldwide. In the present study, a high frequency of Babesia odocoilei, a red blood cell parasite, was observed in the Huronia area of Ontario, Canada. Notably, 71% (15/21) blacklegged ticks, Ixodes scapularis, collected from canine and feline hosts were infected with B. odocoilei. Consistent with U.S. studies, 12.5% (4/32) of questing I. scapularis adults collected by flagging in various parts of southwestern Ontario were positive for B. odocoilei. Our data show that all B. odocoilei strains in the present study have consistent genetic identity, and match type strains in the GenBank database. The high incidence of B. odocoilei in the Huronia area indicates that this babesial infection is established, and is cycling enzootically in the natural environment. Our data confirm that B. odocoilei has wide distribution in southern Ontario.

An HIV-positive man from Zimbabwe living in South Africa sought treatment for multiple clinical signs, including fever, weight loss, anemia, and splenomegaly. We identified in his blood an African rodent piroplasm, Anthemosoma garnhami, related to Babesia species. This finding extends the known geographic and host range of A. garnhami. [ABSTRACT FROM AUTHOR]

Cytauxzoon felis, a tick-borne hemoprotozoal pathogen of felids, causes an acute, often-fatal disease in domestic cats. While public awareness of the disease has increased, few studies have evaluated the incidence of acute cytauxzoonosis cases and their associated risk factors. The objective of this study was to retrospectively review records of cats diagnosed with acute cytauxzoonosis in eastern Kansas from 2006–2019 using clinic records and determine: (i) feline cytauxzoonosis risk factors; and (ii) if cytauxzoonosis case incidence is increasing. Although inter-annual variation of acute cytauxzoonosis diagnosis was observed in the eastern Kansas domestic cat population, the overall incidence trend remained largely unchanged over the 14-year case review period. In comparison to ill (C. felis-unrelated) control cases, more acute cytauxzoonosis cases were diagnosed in spring and summer, suggesting a seasonal fluctuation of infection, with samples most commonly submitted from ≥1 year old, owned, male cats. Although cytauxzoonosis case submissions remained consistent over the broad study period, increasing tick vector and domestic cat reservoir populations may contribute to additional cytauxzoonosis case expansion in endemic areas. Investigating the incidence of acute cytauxzoonosis, patient risk factors, and ecological variables that influence disease transmission are important steps towards developing and communicating the need for effective cytauxzoonosis control strategies for high-risk cat populations.

Tick-borne pathogens cause economically significant diseases in cattle. Theileria spp. are parasitic protozoa and the causative agent of bovine theileriosis. Here we report the distribution and risk factors of bovine Theileria using blood samples taken between 2018 and 2019. Of 737 tested cattle, nine animals (1.2%) were positive for Theileria orientalis infection by 18S rRNA gene amplification. Further analysis of the infected samples using the T. orientalis major piroplasm surface protein (MPSP) gene revealed five different genotypes circulating in the population: Types 1, 2, 3, 7, and N3. To the best of our knowledge, this is the first research to describe the existence of the T. orientalis MPSP genotype N3 in South Korea. Although the prevalence of bovine T. orientalis was low, our study offers data on the geographical distribution and prevalence of bovine Theileria spp. in South Korea. Further studies are warranted to determine the correlation of clinical symptoms with parasite MPSP genotypes. Our data provide epidemiological information to help control bovine theileriosis in South Korea.

Songbirds widely disperse ticks that carry a diversity of pathogens, some of which are pathogenic to humans. Among ticks commonly removed from songbirds, the blacklegged tick, Ixodes scapularis, can harbor any combination of nine zoonotic pathogens, including Babesia species. From May through September 2019, a total 157 ticks were collected from 93 songbirds of 29 species in the Canadian provinces of Ontario and Québec. PCR testing for the 18S gene of Babesia species detected Babesia odocoilei in 12.63% of I. scapularis nymphs parasitizing songbirds in Ontario and Québec; none of the relatively small numbers of Ixodes muris, Ixodes brunneus, or Haemaphysalis leporispalustris were PCR-positive. For ticks at each site, the prevalence of B. odocoilei was 16.67% in Ontario and 8.89% and 5.26% in Québec. Of 31 live, engorged I. scapularis larvae and nymphs held to molt, 25 ticks completed the molt; five of these molted ticks were positive for B. odocoilei. PCR-positive ticks were collected from six bird species—namely, Common Yellowthroat, Swainson’s Thrush, Veery, House Wren, Baltimore Oriole, and American Robin. Phylogenetic analysis documented the close relationship of B. odocoilei to Babesia canis canis and Babesia divergens, the latter a known pathogen to humans. For the first time in Canada, we confirm the transstadial passage of B. odocoilei in I. scapularis molting from larvae to nymphs. A novel host record reveals I. scapularis on a Palm Warbler. Our findings show that B. odocoilei is present in all mobile life stages of I. scapularis, and it is widely dispersed by songbirds in Ontario and Québec.

This is a retrospective study of 38 cases of infection by Babesia macropus, associated with a syndrome of anaemia and debility in hand-reared or free-ranging juvenile eastern grey kangaroos (Macropus giganteus) from coastal New South Wales and south-eastern Queensland between 1995 and 2013. Infection with B. macropus is recorded for the first time in agile wallabies (Macropus agilis) from far north Queensland. Animals in which B. macropus infection was considered to be the primary cause of morbidity had marked anaemia, lethargy and neurological signs, and often died. In these cases, parasitised erythrocytes were few or undetectable in peripheral blood samples but were sequestered in large numbers within small vessels of visceral organs, particularly in the kidney and brain, associated with distinctive clusters of extraerythrocytic organisms. Initial identification of this piroplasm in peripheral blood smears and in tissue impression smears and histological sections was confirmed using transmission electron microscopy and molecular analysis. Samples of kidney, brain or blood were tested using PCR and DNA sequencing of the 18S ribosomal RNA and heat shock protein 70 gene using primers specific for piroplasms. The piroplasm detected in these samples had 100% sequence identity in the 18S rRNA region with the recently described Babesia macropus in two eastern grey kangaroos from New South Wales and Queensland, and a high degree of similarity to an unnamed Babesia sp. recently detected in three woylies (Bettongia penicillata ogilbyi) in Western Australia.

Natural landscape alterations as a consequence of urbanisation are one of the main drivers in the movements of wildlife into metropolitan and peri-urban areas. Worldwide, these wildlife species are highly adaptable and may be responsible for the transmission of tick-borne pathogens including piroplasms ( Babesia , Theileria and Cytauxzoon spp.) that cause piroplasmosis in animals and occasionally in humans. Little is known about piroplasms in the ticks of urban wildlife in Australia. Ticks from long-nosed bandicoots ( Perameles nasuta ; n = 71), eastern-barred bandicoots ( Perameles gunnii ; n = 41), northern-brown bandicoots ( Isoodon macrourus ; n = 19), southern-brown bandicoots ( Isoodon obesulus ; n = 4), bandicoot sp. (n = 2), flying foxes ( Pteropus sp.; n = 3), black rats ( Rattus rattus ; n = 7), bush rats ( Rattus fuscipes ; n = 4), brushtail possums ( Trichosurus vulpecula ; n = 19), ringtail possums ( Pseudocheirus peregrinus ; n = 12), short-eared possums ( Trichosurus caninus ; n = 6), possum sp. ( Trichosurus sp.; n = 8), and red foxes ( Vulpes vulpes ; n = 12) were analysed using piroplasm-specific 18S primers and Sanger sequencing. Seven Ixodes tasmani ticks from long-nosed bandicoots and bandicoots sp., three I. tasmani ticks and one Ixodes holocyclus tick from brushtail possums, and one Haemaphysalis longicornis tick from a red fox were positive for piroplasms. New genotypes, with sequences sharing 98% nucleotide similarities with Theileria sp. K1 detected in a burrowing bettong ( Bettongia lesueur ), were identified from bandicoot ticks. New genotypes were detected in ticks from brushtail possums, which shared 98% similarity with a Babesia sp. (JQ682877) previously identified in marsupials. Theileria orientalis was identified in the H. longicornis tick from the red fox. Babesia and Theileria spp. in the ticks parasitizing bandicoots and brushtail possums clustered closely with respective Babesia and Theileria clades derived from Australian marsupials. This represents the first detection of piroplasms in ticks parasitizing brushtail possums and a red fox in Australia. [ABSTRACT FROM AUTHOR]

Poaching of both black ( Diceros bicornis ) and white ( Ceratotherium simum ) rhinoceros in Africa has increased significantly in recent years. In an effort to ensure the survival of these critically endangered species, breeding programs were established in the 1990s in Australia, where a similar climate and habitat is available. In this study we examined blood samples from two C. simum , including a 16 yr old female (Aluka) who died in captivity, and a 17 yr old asymptomatic male (Umfana). Bloods from seven healthy D. bicornis housed at the zoo were also collected. All samples were tested for the presence of piroplasms via blood smear and PCR. A generic PCR for the 18S rRNA gene of the Piroplasmida revealed the presence of piroplasm infection in both dead and asymptomatic C. simum . Subsequent sequencing of these amplicons revealed the presence of Theileria bicornis . Blood smear indicated that this organism was present at low abundance in both affected and asymptomatic individuals and was not linked to the C. simum mortality. T. bicornis was also detected in the D. bicornis population (n = 7) housed at Taronga Western Plains Zoo using PCR and blood film examination; however only animals imported from Africa (n = 1) tested T. bicornis positive, while captive-born animals bred within Australia (n = 6) tested negative suggesting that transmission within the herd was unlikely. Phylogenetic analysis of the full length T. bicornis 18S rRNA genes classified this organism outside the clade of the transforming and non-transforming Theileria with a new haplotype, H4, identified from D. bicornis . This study revealed the presence of Theileria bicornis in Australian captive populations of both C. simum and D. bicornis and a new haplotype of the parasite was identified. [ABSTRACT FROM AUTHOR]

Babesia and Theileria are intraerythrocytic protozoans of the phylum Apicomplexa. These species are capable of infecting wild and domestic animals and have historically caused great economic loss in the agricultural industry. In recent years human babesiosis has been deemed an emerging zoonosis in North America, Europe and Asia. The first locally acquired case of babesiosis in Australia, caused by Babesia microti , was reported in March 2012. A number of native Babesia and Theileria species have been identified in Australian marsupials, however their associated tick vectors and threat to human health is unknown. In the present study DNA was extracted from 1154 ticks collected from across Australia. PCR was used to amplify a Babesia and Theileria -specific partial region of the 18S ribosomal RNA gene. Positive samples were sequenced and phylogenetic analysis was performed. Twenty-nine sequences were obtained from ticks belonging to the genera Ixodes , Haemaphysalis and Bothriocroton . The sequences were closely related to Babesia macropus, and Theileria recently identified in marsupials and monotremes. Bayesian inference and maximum likelihood methods showed that Australian Babesia and Theileria species form monophyletic groups. [ABSTRACT FROM AUTHOR]