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Leonhard Schnittger - One of the best experts on this subject based on the ideXlab platform.

  • The repertoire of serine rhomboid proteases of piroplasmids of importance to animal and human health.
    International journal for parasitology, 2021
    Co-Authors: Romina Gallenti, Monica Florin-christensen, Tomás Poklepovich, Leonhard Schnittger
    Abstract:

    Abstract Babesia, Theileria and Cytauxzoon are tick-borne apicomplexan protozoans of the order Piroplasmida, notorious for the diseases they cause in livestock, pets and humans. Host cell invasion is their Achilles heel, allowing for the development of drug or vaccine-based therapies. In other apicomplexans, cleavage of the transmembrane domain of adhesins by the serine rhomboid proteinase ROM4 is required for successful completion of invasion. In this study, we record and classify the rhomboid repertoire encoded in the genomes of 10 piroplasmid species pertaining to the lineages Babesia sensu stricto (s.s., Clade VI), Theileria sensu stricto (Clade IV), Theileria equi (Clade IV), Cytauxzoon felis (Clade IIIb) and Babesia microti (Clade I), as defined by Schnittger et al. (2012). Fifty-six piroplasmid rhomboid-like proteins were assigned by phylogenetic analysis and bidirectional best hit to the ROM4, ROM6, ROM7 or ROM8 groups, and their crucial motifs for conformation and function were identified. Forty-four of these rhomboids had either been incorrectly classified or misannotated. Babesia s.s. encode five or three ROM4 proteinase paralogs, whereas the remaining piroplasmids encode two ROM4 paralogs. All piroplasmids encode a single ROM6, ROM7 and ROM8. Thus, an increased paralog number of ROM4 is the only feature distinguishing Babesia s.s. from other piroplasmid lineages. Piroplasmid ROM6 is related to the mammalian mitochondrial rhomboid and, accordingly, N-terminal mitochondrial targeting signal sequence was found in some cases. ROM6 is the only rhomboid encoded by piroplasmids that is ubiquitous in other organisms. ROM8 represents a pseudoproteinase that is highly conserved between studied piroplasmids, suggesting that it important in regulatory function. ROM4, ROM6, ROM7 and ROM8 are exclusively present in Aconoidasida, which comprises piroplasmids and Plasmodium, suggesting a relevant functional role in erythrocyte invasion. The correct classification and designation of piroplasmid rhomboids presented in this study facilitates an informed choice for future in-depth study of their functions.

  • N-Glycosylation in Piroplasmids: Diversity within Simplicity
    Pathogens (Basel Switzerland), 2021
    Co-Authors: Monica Florin-christensen, Anabel Rodriguez, Carlos E. Suarez, Massaro W. Ueti, Fernando Oscar Delgado, Ignacio Echaide, Leonhard Schnittger
    Abstract:

    N-glycosylation has remained mostly unexplored in Piroplasmida, an order of tick-transmitted pathogens of veterinary and medical relevance. Analysis of 11 piroplasmid genomes revealed three distinct scenarios regarding N-glycosylation: Babesia sensu stricto (s.s.) species add one or two N-acetylglucosamine (NAcGlc) molecules to proteins; Theileria equi and Cytauxzoon felis add (NAcGlc)2-mannose, while B. microti and Theileria s.s. synthesize dolichol-P-P-NAcGlc and dolichol-P-P-(NAcGlc)2 without subsequent transfer to proteins. All piroplasmids possess the gene complement needed for the synthesis of the N-glycosylation substrates, dolichol-P and sugar nucleotides. The oligosaccharyl transferase of Babesia species, T. equi and C. felis, is predicted to be composed of only two subunits, STT3 and Ost1. Occurrence of short N-glycans in B. bovis merozoites was experimentally demonstrated by fluorescence microscopy using a NAcGlc-specific lectin. In vitro growth of B. bovis was significantly impaired by tunicamycin, an inhibitor of N-glycosylation, indicating a relevant role for N-glycosylation in this pathogen. Finally, genes coding for N-glycosylation enzymes and substrate biosynthesis are transcribed in B. bovis blood and tick stages, suggesting that this pathway is biologically relevant throughout the parasite life cycle. Elucidation of the role/s exerted by N-glycans will increase our understanding of these successful parasites, for which improved control measures are needed.

  • A new piroplasmid species infecting dogs: morphological and molecular characterization and pathogeny of Babesia negevi n. sp.
    Parasites & Vectors, 2020
    Co-Authors: Gad Baneth, Leonhard Schnittger, Adam J. Birkenheuer, Megan E. Schreeg, Monica Florin-christensen, Yaarit Nachum-biala, Hagar Prince, Itamar Aroch
    Abstract:

    Introduction Babesiosis is a protozoan tick-borne infection associated with anemia and life-threatening disease in humans, domestic and wildlife animals. Dogs are infected by at least six well-characterized Babesia spp. that cause clinical disease. Infection with a piroplasmid species was detected by light microscopy of stained blood smears from five sick dogs from Israel and prompted an investigation on the parasite’s identity. Methods Genetic characterization of the piroplasmid was performed by PCR amplification of the 18S rRNA and the cytochrome c oxidase subunit 1 ( cox 1) genes, DNA sequencing and phylogenetic analysis. Four of the dogs were co-infected with Borrelia persica (Dschunkowsky, 1913), a relapsing fever spirochete transmitted by the argasid tick Ornithodoros tholozani Laboulbène & Mégnin. Co-infection of dogs with B. persica raised the possibility of transmission by O. tholozani and therefore, a piroplasmid PCR survey of ticks from this species was performed. Results The infected dogs presented with fever (4/5), anemia, thrombocytopenia (4/5) and icterus (3/5). Comparison of the 18S rRNA and cox 1 piroplasmid gene sequences revealed 99–100% identity between sequences amplified from different dogs and ticks. Phylogenetic trees demonstrated a previously undescribed species of Babesia belonging to the western group of Babesia ( sensu lato ) and closely related to the human pathogen Babesia duncani Conrad, Kjemtrup, Carreno, Thomford, Wainwright, Eberhard, Quick, Telfrom & Herwalt, 2006 while more moderately related to Babesia conradae Kjemtrup, Wainwright, Miller, Penzhorn & Carreno, 2006 which infects dogs. The piroplasm forms detected included tetrads (Maltese cross), merozoite and trophozoite stages whose average size was larger than stages of other canine Babesia spp. belonging to the Babesia ( s.l .) and B. gibsoni Patton, 1910, and smaller than other canine Babesia ( sensu stricto ) spp. Of 212 O. tholozani ticks surveyed, 11 (5.2%) harbored DNA of the new species of Babesia . Conclusions Babesia negevi n. sp. is described based on morphological and genetic characterization and phylogenetic analyses. The species is named after the Negev desert of southern Israel, where the first infected dog originated from. Despite co-infection in four dogs, the fifth dog had fatal disease attesting that B. negevi n. sp. infection requires clinical attention. Incriminating O. tholozani or another tick species as the vector of Babesia negevi n. sp., would require additional studies.

  • Babesia life cycle – When phylogeny meets biology
    Trends in parasitology, 2019
    Co-Authors: Marie Jalovecka, Daniel Sojka, Mariano E. Ascencio, Leonhard Schnittger
    Abstract:

    Although Babesia represents an important worldwide veterinary threat and an emerging risk to humans, this parasite has been poorly studied as compared to Plasmodium, its malaria-causing relative. In fact, Babesia employs highly specific survival strategies during its intraerythrocytic development and its intricate journey through the tick vector. This review introduces a substantially extended molecular phylogeny of the order Piroplasmida, challenging previous taxonomic classifications. The intriguing developmental proficiencies of Babesia are highlighted and compared with those of other haemoparasitic Apicomplexa. Molecular mechanisms associated with distinctive events in the Babesia life cycle are emphasized as potential targets for the development of Babesia-specific treatments.

  • Cysteine Proteinase C1A Paralog Profiles Correspond with Phylogenetic Lineages of Pathogenic Piroplasmids
    MDPI AG, 2018
    Co-Authors: Mariano E. Ascencio, Monica Florin-christensen, Choukri B. Mamoun, William Weir, Brian Shiels, Leonhard Schnittger
    Abstract:

    Piroplasmid parasites comprising of Babesia, Theileria, and Cytauxzoon are transmitted by ticks to farm and pet animals and have a significant impact on livestock industries and animal health in tropical and subtropical regions worldwide. In addition, diverse Babesia spp. infect humans as opportunistic hosts. Molecular phylogeny has demonstrated at least six piroplasmid lineages exemplified by B. microti, B. duncani, C. felis, T. equi, Theileria sensu stricto (T. annulata, T. parva, and T. orientalis) and Babesia sensu stricto (B. bovis, B. bigemina, and B. ovis). C1A cysteine-proteinases (C1A-Cp) are papain-like enzymes implicated in pathogenic and vital steps of the parasite life cycle such as nutrition and host cell egress. An expansion of C1A-Cp of T. annulata and T. parva with respect to B. bovis and B. ovis was previously described. In the present work, C1A-Cp paralogs were identified in available genomes of species pertaining to each piroplasmid lineage. Phylogenetic analysis revealed eight C1A-Cp groups. The profile of C1A-Cp paralogs across these groups corroborates and defines the existence of six piroplasmid lineages. C. felis, T. equi and Theileria s.s. each showed characteristic expansions into extensive families of C1A-Cp paralogs in two of the eight groups. Underlying gene duplications have occurred as independent unique evolutionary events that allow distinguishing these three piroplasmid lineages. We hypothesize that C1A-Cp paralog families may be associated with the advent of the schizont stage. Differences in the invertebrate tick host specificity and/or mode of transmission in piroplasmid lineages might also be associated with the observed C1A-Cp paralog profiles

Gad Baneth - One of the best experts on this subject based on the ideXlab platform.

  • A new piroplasmid species infecting dogs: morphological and molecular characterization and pathogeny of Babesia negevi n. sp.
    Parasites & Vectors, 2020
    Co-Authors: Gad Baneth, Leonhard Schnittger, Adam J. Birkenheuer, Megan E. Schreeg, Monica Florin-christensen, Yaarit Nachum-biala, Hagar Prince, Itamar Aroch
    Abstract:

    Introduction Babesiosis is a protozoan tick-borne infection associated with anemia and life-threatening disease in humans, domestic and wildlife animals. Dogs are infected by at least six well-characterized Babesia spp. that cause clinical disease. Infection with a piroplasmid species was detected by light microscopy of stained blood smears from five sick dogs from Israel and prompted an investigation on the parasite’s identity. Methods Genetic characterization of the piroplasmid was performed by PCR amplification of the 18S rRNA and the cytochrome c oxidase subunit 1 ( cox 1) genes, DNA sequencing and phylogenetic analysis. Four of the dogs were co-infected with Borrelia persica (Dschunkowsky, 1913), a relapsing fever spirochete transmitted by the argasid tick Ornithodoros tholozani Laboulbène & Mégnin. Co-infection of dogs with B. persica raised the possibility of transmission by O. tholozani and therefore, a piroplasmid PCR survey of ticks from this species was performed. Results The infected dogs presented with fever (4/5), anemia, thrombocytopenia (4/5) and icterus (3/5). Comparison of the 18S rRNA and cox 1 piroplasmid gene sequences revealed 99–100% identity between sequences amplified from different dogs and ticks. Phylogenetic trees demonstrated a previously undescribed species of Babesia belonging to the western group of Babesia ( sensu lato ) and closely related to the human pathogen Babesia duncani Conrad, Kjemtrup, Carreno, Thomford, Wainwright, Eberhard, Quick, Telfrom & Herwalt, 2006 while more moderately related to Babesia conradae Kjemtrup, Wainwright, Miller, Penzhorn & Carreno, 2006 which infects dogs. The piroplasm forms detected included tetrads (Maltese cross), merozoite and trophozoite stages whose average size was larger than stages of other canine Babesia spp. belonging to the Babesia ( s.l .) and B. gibsoni Patton, 1910, and smaller than other canine Babesia ( sensu stricto ) spp. Of 212 O. tholozani ticks surveyed, 11 (5.2%) harbored DNA of the new species of Babesia . Conclusions Babesia negevi n. sp. is described based on morphological and genetic characterization and phylogenetic analyses. The species is named after the Negev desert of southern Israel, where the first infected dog originated from. Despite co-infection in four dogs, the fifth dog had fatal disease attesting that B. negevi n. sp. infection requires clinical attention. Incriminating O. tholozani or another tick species as the vector of Babesia negevi n. sp., would require additional studies.

  • Epidemiological Study of Canine Babesiosis and Hepatozoonosis in the South of Romania
    Acta Parasitologica, 2020
    Co-Authors: Andrei Alexandru Cimpan, Yaarit Nachum-biala, Bar Ben-shitrit, Liviu Miron, Gad Baneth
    Abstract:

    Purpose The aim of this study was to evaluate the prevalence and distribution of canine babesiosis and hepatozoonosis in the south of Romania as well as to assess if there are differences in the prevalence between regions in that area. Methods Blood samples from 300 dogs from the south of Romania were screened for the presence of piroplasmids by PCR. Positive samples were further tested for the presence of Babesia spp . and Hepatozoon canis . Pearson's chi-square with Yates correction was used to determine if there is statistical difference between infected populations. Results Fifty-four percent (163/300) of the dogs were positive for Hepatozoon spp. or piroplasmids. Forty-eight percent (143/300) were infected with H. canis and 9.6% (29/300) were positive to B. canis . Three percent (9/300) of the dogs were co-infected with Babesia spp. and H. canis . The prevalence of H. canis in the south-west of Romania was significantly higher ( p  

  • epidemiological study of canine babesiosis and hepatozoonosis in the south of romania
    Acta Parasitologica, 2020
    Co-Authors: Andrei Alexandru Cimpan, Liviu Miron, Yaarit Nachumbiala, Bar Benshitrit, Gad Baneth
    Abstract:

    PURPOSE The aim of this study was to evaluate the prevalence and distribution of canine babesiosis and hepatozoonosis in the south of Romania as well as to assess if there are differences in the prevalence between regions in that area. METHODS Blood samples from 300 dogs from the south of Romania were screened for the presence of piroplasmids by PCR. Positive samples were further tested for the presence of Babesia spp. and Hepatozoon canis. Pearson's chi-square with Yates correction was used to determine if there is statistical difference between infected populations. RESULTS Fifty-four percent (163/300) of the dogs were positive for Hepatozoon spp. or piroplasmids. Forty-eight percent (143/300) were infected with H. canis and 9.6% (29/300) were positive to B. canis. Three percent (9/300) of the dogs were co-infected with Babesia spp. and H. canis. The prevalence of H. canis in the south-west of Romania was significantly higher (p < 0.05) than that of three other regions while the prevalence of Babesia spp. infection in the south-east of the country was significantly higher (p < 0.05) compared to its south-west. CONCLUSIONS A high rate of H. canis infection was found in the stray dog population surveyed with the highest infection rate detected in south-western Romania. Babesia canis was found to pose the highest threat in south-eastern Romania, whereas B. vogeli infection was focused in the same region of high H. canis infection in south-western Romania. These infections should be of important concern to veterinarians and dog owners in Romania.

  • reclassification of theileria annae as babesia vulpes sp nov
    Parasites & Vectors, 2015
    Co-Authors: Gad Baneth, Leonhard Schnittger, Luis Cardoso, Monica Florinchristensen
    Abstract:

    Background Theileria annae is a tick-transmitted small piroplasmid that infects dogs and foxes in North America and Europe. Due to disagreement on its placement in the Theileria or Babesia genera, several synonyms have been used for this parasite, including Babesia Spanish dog isolate, Babesia microti-like, Babesia (Theileria) annae, and Babesia cf. microti. Infections by this parasite cause anemia, thrombocytopenia, and azotemia in dogs but are mostly subclinical in red foxes (Vulpes vulpes). Furthermore, high infection rates have been detected among red fox populations in distant regions strongly suggesting that these canines act as the parasite’s natural host. This study aims to reassess and harmonize the phylogenetic placement and binomen of T. annae within the order Piroplasmida.

  • Reclassification of Theileria annae as Babesia vulpes sp. nov.
    Parasites & Vectors, 2015
    Co-Authors: Gad Baneth, Luis Cardoso, Monica Florin-christensen, Leonhard Schnittger
    Abstract:

    Background Theileria annae is a tick-transmitted small piroplasmid that infects dogs and foxes in North America and Europe. Due to disagreement on its placement in the Theileria or Babesia genera, several synonyms have been used for this parasite, including Babesia Spanish dog isolate, Babesia microti -like, Babesia ( Theileria ) annae , and Babesia cf. microti . Infections by this parasite cause anemia, thrombocytopenia, and azotemia in dogs but are mostly subclinical in red foxes ( Vulpes vulpes ). Furthermore, high infection rates have been detected among red fox populations in distant regions strongly suggesting that these canines act as the parasite’s natural host. This study aims to reassess and harmonize the phylogenetic placement and binomen of T. annae within the order Piroplasmida. Methods Four molecular phylogenetic trees were constructed using a maximum likelihood algorithm based on DNA alignments of: (i) near-complete 18S rRNA gene sequences (n = 76 and n = 93), (ii) near-complete and incomplete 18S rRNA gene sequences (n = 92), and (iii) tubulin-beta gene sequences (n = 32) from B. microti and B. microti -related parasites including those detected in dogs and foxes. Results All phylogenetic trees demonstrate that T. annae and its synonyms are not Theileria parasites but are most closely related with B. microti . The phylogenetic tree based on the 18S rRNA gene forms two separate branches with high bootstrap value, of which one branch corresponds to Babesia species infecting rodents, humans, and macaques, while the other corresponds to species exclusively infecting carnivores. Within the carnivore group, T. annae and its synonyms from distant regions segregate into a single clade with a highly significant bootstrap value corroborating their separate species identity. Conclusion Phylogenetic analysis clearly shows that T. annae and its synonyms do not pertain to Theileria and can be clearly defined as a separate species. Based on the facts that T. annae and its synonyms have not been shown to have a leukocyte stage, as expected in Theileria , do not infect humans and rodents as B. microti , and cluster phylogenetically as a separate species, this study proposes to name this parasite Babesia vulpes sp. nov., after its natural host, the red fox V. vulpes .

Javier Millan - One of the best experts on this subject based on the ideXlab platform.

  • Babesia vogeli in dogs in Chile.
    Transboundary and emerging diseases, 2020
    Co-Authors: Sophia Di Cataldo, Claudia Ulloa‐contreras, Aitor Cevidanes, Claudia Hernández, Javier Millan
    Abstract:

    We report the presence of Babesia vogeli in dogs in Chile. During two surveillance campaigns separated by a year, sixty-three blood samples from free-ranging rural dogs in Coquimbo, Chile, were analysed through conventional PCR screening of the 18S rRNA for Babesia species. Sequencing confirmed the presence of Babesia vogeli in 6.3% of the tested animals, with positive cases in both years. All the sequences showed 100% nucleotide sequence identity among them and 99.8% with other previously published sequences from dogs. No clinical signs or haematological abnormalities other than thrombocytopenia were found in the parasitized individuals. This is the first report of a canine piroplasmid in Chile.

  • Molecular investigation of vector-borne parasites in wild micromammals, Barcelona (Spain)
    Parasitology Research, 2018
    Co-Authors: Javier Millan
    Abstract:

    Information about the role of micromammals in the epidemiology of Leishmania infantum and Babesia sp. in southern Europe is scarce. Wild micromammals were captured in natural areas in the Barcelona province (NE Spain) in 2011 and analysed by real-time PCR for the presence of DNA of Piroplasmida ( n  = 253) and Leishmania infantum ( n  = 166). The wood mouse ( Apodemus sylvaticus ) was the most abundant species, but all specimens tested negative for L. infantum , for which two out of 15 (13.3%) white-toothed shrews ( Crocidura russula ) and one out of 23 (4.3%) Algerian mice ( Mus spretus ) resulted positive. No individual was positive for Piroplasmida DNA. The role of wild micromammals in the epidemiology of these parasites in the study area seems negligible.

  • a review of piroplasmid infections in wild carnivores worldwide importance for domestic animal health and wildlife conservation
    Parasites & Vectors, 2016
    Co-Authors: Mario Alvaradorybak, Laia Solanogallego, Javier Millan
    Abstract:

    Piroplasmids are tick-borne protozoan parasites that infect blood cells (erythrocytes, lymphocytes or other leukocytes) or endothelial cells of numerous wild and domestic vertebrates worldwide. They cause severe disease in livestock, dogs, cats, wild mammals and, occasionally, in humans. Piroplasmid infections are prevalent in wild carnivores worldwide although there is limited information about their clinical and epidemiological importance. There are currently nine recognized species of Babesia, two of Theileria, two of Cytauxzoon and one of Rangelia infecting captive and wild carnivores, including members of Canidae, Felidae, Mustelidae, Procyonidae, Ursidae, Viverridae, Hyaenidae and Herpestidae in the Americas, Eurasia and Africa. However, the number of piroplasmid species is likely higher than currently accepted due to the reported existence of DNA sequences that may correspond to new species and the lack of studies on many host species and biogeographical areas. Indeed, many species have been recognized in the last few years with the advancement of molecular analyses. Disease and mortality have been documented in some wild carnivores, whereas other species appear to act as natural, subclinical reservoirs. Various factors (e.g. unnatural hosts, stress due to captivity, habitat degradation, climate fluctuation or immunosuppression) have been associated with disease susceptibility to piroplasmid infections in some species in captivity. We aimed to review the current knowledge on the epidemiology of piroplasmid infections in wild carnivores and associated tick vectors. Emphasis is given to the role of wild carnivores as reservoirs of clinical piroplasmosis for domestic dogs and cats, and to the importance of piroplasmids as disease agents for endangered carnivores.

  • Molecular detection of vector-borne pathogens in wild and domestic carnivores and their ticks at the human–wildlife interface
    Ticks and tick-borne diseases, 2015
    Co-Authors: Javier Millan, Jose De La Fuente, Tatiana Proboste, Isabel G. Fernández De Mera, Andrea D. Chirife, Laura Altet
    Abstract:

    Urbanization of natural areas is considered one of the causes of the current apparent emergence of infectious diseases. Carnivores are among the species that adapt well to urban and periurban environments, facilitating cross-species disease transmission with domestic dogs and cats, and potentially with their owners. The prevalence of vector-borne pathogens (VBP) of zoonotic and veterinary interest was studied in sympatric wild and domestic carnivores into Barcelona Metropolitan Area (NE Spain). Blood or spleen samples from 130 animals, including 34 common genets (Genetta genetta), 12 red foxes (Vulpes vulpes), 10 stone martens (Martes foina), three Eurasian badgers (Meles meles), 34 free-roaming domestic cats and 37 dogs with outdoor access, were collected either in protected or adjacent residential areas. A total of 309 ticks (chiefly Rhipicephalus turanicus) were collected on these animals. The samples were analyzed with a battery of PCR assays targeting the DNA of Rickettsia spp., Anaplasmataceae, Coxiella burnetii, Bartonella spp., and Piroplasmida, and the amplicons were sequenced. The fox showed the highest prevalence (58%) and diversity of VBP (four pathogens), whereas none of the dogs were infected. Bartonella spp. (including B. clarridgeiae, B. henselae, and B. rochalimae) was the most prevalent pathogen. Infection of wild carnivores with Ehrlichia canis, C. burnetii, Theileria annae and Babesia vogeli was also confirmed, with some cases of coinfection observed. The presence of DNA of T. annae and B. vogeli was also confirmed in tick pools from four species of wild carnivores, supporting their role in piroplasmid life-cycle. By the sequencing of several target genes, DNA of Rickettsia massiliae was confirmed in 17 pools of Rh. turanicus, Rh. sanguineous, and Rh. pusillus from five different species, and Rickettsia conorii in one pool of Rh. sanguineous from a dog. None of the hosts from which these ticks were collected was infected by Rickettsia. Although carnivores may not be reservoir hosts for zoonotic Rickettsia, they can have an important role as mechanical dispersers of infected ticks.

David Modrý - One of the best experts on this subject based on the ideXlab platform.

  • a molecular survey on host specificity of feline and canine hepatozoon in model site of northern kenya
    Parasites & Vectors, 2014
    Co-Authors: Moneeb Ahmad Qablan, David Modrý, Gad Baneth, Martina Gallusova, A D Mihalca
    Abstract:

    Species of the genus Hepatozoon are apicomplexan parasites transmitted by variety of hematophagous arthropods to a wide range of intermediate hosts. Hepatozoon infections are broadly distributed around the world and the presence of this parasite is confirmed both in cats and dogs. Until now, two species from canines (H. canis, H. americanum) and a single species from felines (H. felis) have been reported. While the canine Hepatozoon spp. are transmitted by ticks, the vector for H. felis remains unknown. The aim of presented study was to determine the prevalence and the diversity of Hepatozoon in population of domestic cats and dogs living in close contact in rural communities of Samburu pastoralists in northern Kenya and to evaluate the existence of possible cross-transmissions between both hosts. Between years 2007-2012 in total, 135 and 258 blood samples from cats and dogs from the area of Mt. Kulal (5 localities) was collected, respectively. The DNA from blood (preserved in ethanol) was extracted using phenol-chlorophorm method and followed by conventional PCR screening. First round of PCR was done by using Piroplasmid-F and Piroplasmid-R primers, amplifying 18S rRNA gene of Hepatozoon spp. (400 bp). A second PCR assay was performed in samples positive by Piroplasmid primers to amplify a larger fragment (1400 bp) of the 18S rRNA gene. All PCR amplicons comming from positive samples were sequenced and determined according to the BLAST match. In cats, this revealed 110 out of 135 (81.5%) positive samples, of which we obtained 104 sequences with an identity of 98% -100% to an existing GenBank accession. In total, 77 sequences were confirmed as Hepatozoon felis, 26 samples was proven to be Hepatozoon sp. and a single sample was determined as Hepatozoon canis with an identity of 96%. Regarding dogs, 121 out of 258 (47%) were positive and we gained 107 sequences of which 105 showed similarity to Hepatozoon canis and the remaining two proved to be Hepatozoon sp. Despite a close contact of hosts, obviously cats and dogs do not share the same parasite, which demonstrates a great majority of clearly identified Hepatozoon canis/Hepatozoon felis. Mentioned facts could be explained either by strict host specificity or by presence of different vector or by both statements.

  • Infections by Babesia caballi and Theileria equi in Jordanian equids: epidemiology and genetic diversity
    Parasitology, 2013
    Co-Authors: Moneeb Ahmad Qablan, Michal Sloboda, Miroslav Oborník, Petr Hořín, Julius Lukeš, Klára J. Petrželková, Mustafa F. Shudiefat, David Modrý
    Abstract:

    Microscopic diagnosis of equine piroplasmoses, caused by Theileria equi and Babesia caballi , is hindered by low parasitaemia during the latent phase of the infections. However, this constraint can be overcome by the application of PCR followed by sequencing. Out of 288 animals examined, the piroplasmid DNA was detected in 78 (27·1%). Multiplex PCR indicated that T. equi (18·8%) was more prevalent than B. caballi (7·3%), while mixed infections were conspicuously absent. Sequences of 69 PCR amplicons obtained by the ‘catch-all’ PCR were in concordance with those amplified by the multiplex strategy. Computed minimal adequate model analyses for both equine piroplasmid species separately showed a significant effect of host species and age in the case of T. equi , while in the B. caballi infections only the correlation with host sex was significant. Phylogenetic analyses inferred the occurrence of three genotypes of T. equi and B. caballi . Moreover, a novel genotype C of B. caballi was identified. The dendrogram based on obtained sequences of T. equi revealed possible speciation events. The infections with T. equi and B. caballi are enzootic in all ecozones of Jordan and different genotypes circulate wherever dense horse population exists.

  • Stray dogs of northern Jordan as reservoirs of ticks and tick-borne hemopathogens
    Parasitology Research, 2012
    Co-Authors: Moneeb Ahmad Qablan, David Modrý, Michaela Kubelová, Pavel Široký, Zuhair Sami Amr
    Abstract:

    Dogs are competent reservoir hosts of several hemopathogens including zoonotic agents and can serve as readily available source of nutrition for many blood-feeding arthropods. Three hemopathogens had been detected for the first time in Jordan. The PCR prevalence of Anaplasma phagocytophilum , Hepatozoon canis , and piroplasmid DNA were 39.5%, 28.9%, and 7.9% ( n  = 38) respectively. Sequencing of amplicons of PCR with universal primers targeting the 18S rRNA gene of piroplasmids shows the highest similarity to equine piroplasmids species Theileria equi from two dogs and Babesia caballi from a single dog. Ticks of two genera Rhipicephalus and Haemaphysalis , were detected in this study ( n  = 268). The brown dog tick, Rhipicephalus sanguineus was the most abundant species (95.1%, n  = 255), followed by Haemaphysalis erinacei (3%, n  = 8) and Haemaphysalis parva (1.9%, n  = 5). The two Haemaphysalis species were detected for the first time from dogs in Jordan. Regarding its high prevalence, we expect R. sanguineus being a possible vector of detected pathogens.

  • Quest for the piroplasms in camels: identification of Theileria equi and Babesia caballi in Jordanian dromedaries by PCR.
    Veterinary parasitology, 2011
    Co-Authors: Moneeb Ahmad Qablan, Michal Sloboda, Milan Jirků, Miroslav Oborník, Samir Dwairi, Zuhair Sami Amr, Petr Hořín, Julius Lukeš, David Modrý
    Abstract:

    DNA of two species of piroplasmids was detected in dromedaries during a survey of blood protozoans in Jordan between 2007 and 2009. Ten clinically healthy camels (10%) originating from three Jordanian districts were found, using a PCR assay, to harbor Theileria or Babesia species in their blood and no mix infection was determined. Analysis of the partial 18S rRNA gene sequences of these parasites allowed their unambiguous identification as equine piroplasmids Babesia caballi (n=6) and Theileria equi (n=4). In case of latter species, a novel genotype was found in horses. This first molecular-based species determination of piroplasmids from camels further contributes to the growing evidence of low host specificity of piroplasmids.

Monica Florin-christensen - One of the best experts on this subject based on the ideXlab platform.

  • The repertoire of serine rhomboid proteases of piroplasmids of importance to animal and human health.
    International journal for parasitology, 2021
    Co-Authors: Romina Gallenti, Monica Florin-christensen, Tomás Poklepovich, Leonhard Schnittger
    Abstract:

    Abstract Babesia, Theileria and Cytauxzoon are tick-borne apicomplexan protozoans of the order Piroplasmida, notorious for the diseases they cause in livestock, pets and humans. Host cell invasion is their Achilles heel, allowing for the development of drug or vaccine-based therapies. In other apicomplexans, cleavage of the transmembrane domain of adhesins by the serine rhomboid proteinase ROM4 is required for successful completion of invasion. In this study, we record and classify the rhomboid repertoire encoded in the genomes of 10 piroplasmid species pertaining to the lineages Babesia sensu stricto (s.s., Clade VI), Theileria sensu stricto (Clade IV), Theileria equi (Clade IV), Cytauxzoon felis (Clade IIIb) and Babesia microti (Clade I), as defined by Schnittger et al. (2012). Fifty-six piroplasmid rhomboid-like proteins were assigned by phylogenetic analysis and bidirectional best hit to the ROM4, ROM6, ROM7 or ROM8 groups, and their crucial motifs for conformation and function were identified. Forty-four of these rhomboids had either been incorrectly classified or misannotated. Babesia s.s. encode five or three ROM4 proteinase paralogs, whereas the remaining piroplasmids encode two ROM4 paralogs. All piroplasmids encode a single ROM6, ROM7 and ROM8. Thus, an increased paralog number of ROM4 is the only feature distinguishing Babesia s.s. from other piroplasmid lineages. Piroplasmid ROM6 is related to the mammalian mitochondrial rhomboid and, accordingly, N-terminal mitochondrial targeting signal sequence was found in some cases. ROM6 is the only rhomboid encoded by piroplasmids that is ubiquitous in other organisms. ROM8 represents a pseudoproteinase that is highly conserved between studied piroplasmids, suggesting that it important in regulatory function. ROM4, ROM6, ROM7 and ROM8 are exclusively present in Aconoidasida, which comprises piroplasmids and Plasmodium, suggesting a relevant functional role in erythrocyte invasion. The correct classification and designation of piroplasmid rhomboids presented in this study facilitates an informed choice for future in-depth study of their functions.

  • N-Glycosylation in Piroplasmids: Diversity within Simplicity
    Pathogens (Basel Switzerland), 2021
    Co-Authors: Monica Florin-christensen, Anabel Rodriguez, Carlos E. Suarez, Massaro W. Ueti, Fernando Oscar Delgado, Ignacio Echaide, Leonhard Schnittger
    Abstract:

    N-glycosylation has remained mostly unexplored in Piroplasmida, an order of tick-transmitted pathogens of veterinary and medical relevance. Analysis of 11 piroplasmid genomes revealed three distinct scenarios regarding N-glycosylation: Babesia sensu stricto (s.s.) species add one or two N-acetylglucosamine (NAcGlc) molecules to proteins; Theileria equi and Cytauxzoon felis add (NAcGlc)2-mannose, while B. microti and Theileria s.s. synthesize dolichol-P-P-NAcGlc and dolichol-P-P-(NAcGlc)2 without subsequent transfer to proteins. All piroplasmids possess the gene complement needed for the synthesis of the N-glycosylation substrates, dolichol-P and sugar nucleotides. The oligosaccharyl transferase of Babesia species, T. equi and C. felis, is predicted to be composed of only two subunits, STT3 and Ost1. Occurrence of short N-glycans in B. bovis merozoites was experimentally demonstrated by fluorescence microscopy using a NAcGlc-specific lectin. In vitro growth of B. bovis was significantly impaired by tunicamycin, an inhibitor of N-glycosylation, indicating a relevant role for N-glycosylation in this pathogen. Finally, genes coding for N-glycosylation enzymes and substrate biosynthesis are transcribed in B. bovis blood and tick stages, suggesting that this pathway is biologically relevant throughout the parasite life cycle. Elucidation of the role/s exerted by N-glycans will increase our understanding of these successful parasites, for which improved control measures are needed.

  • A new piroplasmid species infecting dogs: morphological and molecular characterization and pathogeny of Babesia negevi n. sp.
    Parasites & Vectors, 2020
    Co-Authors: Gad Baneth, Leonhard Schnittger, Adam J. Birkenheuer, Megan E. Schreeg, Monica Florin-christensen, Yaarit Nachum-biala, Hagar Prince, Itamar Aroch
    Abstract:

    Introduction Babesiosis is a protozoan tick-borne infection associated with anemia and life-threatening disease in humans, domestic and wildlife animals. Dogs are infected by at least six well-characterized Babesia spp. that cause clinical disease. Infection with a piroplasmid species was detected by light microscopy of stained blood smears from five sick dogs from Israel and prompted an investigation on the parasite’s identity. Methods Genetic characterization of the piroplasmid was performed by PCR amplification of the 18S rRNA and the cytochrome c oxidase subunit 1 ( cox 1) genes, DNA sequencing and phylogenetic analysis. Four of the dogs were co-infected with Borrelia persica (Dschunkowsky, 1913), a relapsing fever spirochete transmitted by the argasid tick Ornithodoros tholozani Laboulbène & Mégnin. Co-infection of dogs with B. persica raised the possibility of transmission by O. tholozani and therefore, a piroplasmid PCR survey of ticks from this species was performed. Results The infected dogs presented with fever (4/5), anemia, thrombocytopenia (4/5) and icterus (3/5). Comparison of the 18S rRNA and cox 1 piroplasmid gene sequences revealed 99–100% identity between sequences amplified from different dogs and ticks. Phylogenetic trees demonstrated a previously undescribed species of Babesia belonging to the western group of Babesia ( sensu lato ) and closely related to the human pathogen Babesia duncani Conrad, Kjemtrup, Carreno, Thomford, Wainwright, Eberhard, Quick, Telfrom & Herwalt, 2006 while more moderately related to Babesia conradae Kjemtrup, Wainwright, Miller, Penzhorn & Carreno, 2006 which infects dogs. The piroplasm forms detected included tetrads (Maltese cross), merozoite and trophozoite stages whose average size was larger than stages of other canine Babesia spp. belonging to the Babesia ( s.l .) and B. gibsoni Patton, 1910, and smaller than other canine Babesia ( sensu stricto ) spp. Of 212 O. tholozani ticks surveyed, 11 (5.2%) harbored DNA of the new species of Babesia . Conclusions Babesia negevi n. sp. is described based on morphological and genetic characterization and phylogenetic analyses. The species is named after the Negev desert of southern Israel, where the first infected dog originated from. Despite co-infection in four dogs, the fifth dog had fatal disease attesting that B. negevi n. sp. infection requires clinical attention. Incriminating O. tholozani or another tick species as the vector of Babesia negevi n. sp., would require additional studies.

  • Cysteine Proteinase C1A Paralog Profiles Correspond with Phylogenetic Lineages of Pathogenic Piroplasmids
    MDPI AG, 2018
    Co-Authors: Mariano E. Ascencio, Monica Florin-christensen, Choukri B. Mamoun, William Weir, Brian Shiels, Leonhard Schnittger
    Abstract:

    Piroplasmid parasites comprising of Babesia, Theileria, and Cytauxzoon are transmitted by ticks to farm and pet animals and have a significant impact on livestock industries and animal health in tropical and subtropical regions worldwide. In addition, diverse Babesia spp. infect humans as opportunistic hosts. Molecular phylogeny has demonstrated at least six piroplasmid lineages exemplified by B. microti, B. duncani, C. felis, T. equi, Theileria sensu stricto (T. annulata, T. parva, and T. orientalis) and Babesia sensu stricto (B. bovis, B. bigemina, and B. ovis). C1A cysteine-proteinases (C1A-Cp) are papain-like enzymes implicated in pathogenic and vital steps of the parasite life cycle such as nutrition and host cell egress. An expansion of C1A-Cp of T. annulata and T. parva with respect to B. bovis and B. ovis was previously described. In the present work, C1A-Cp paralogs were identified in available genomes of species pertaining to each piroplasmid lineage. Phylogenetic analysis revealed eight C1A-Cp groups. The profile of C1A-Cp paralogs across these groups corroborates and defines the existence of six piroplasmid lineages. C. felis, T. equi and Theileria s.s. each showed characteristic expansions into extensive families of C1A-Cp paralogs in two of the eight groups. Underlying gene duplications have occurred as independent unique evolutionary events that allow distinguishing these three piroplasmid lineages. We hypothesize that C1A-Cp paralog families may be associated with the advent of the schizont stage. Differences in the invertebrate tick host specificity and/or mode of transmission in piroplasmid lineages might also be associated with the observed C1A-Cp paralog profiles

  • Reclassification of Theileria annae as Babesia vulpes sp. nov.
    Parasites & Vectors, 2015
    Co-Authors: Gad Baneth, Luis Cardoso, Monica Florin-christensen, Leonhard Schnittger
    Abstract:

    Background Theileria annae is a tick-transmitted small piroplasmid that infects dogs and foxes in North America and Europe. Due to disagreement on its placement in the Theileria or Babesia genera, several synonyms have been used for this parasite, including Babesia Spanish dog isolate, Babesia microti -like, Babesia ( Theileria ) annae , and Babesia cf. microti . Infections by this parasite cause anemia, thrombocytopenia, and azotemia in dogs but are mostly subclinical in red foxes ( Vulpes vulpes ). Furthermore, high infection rates have been detected among red fox populations in distant regions strongly suggesting that these canines act as the parasite’s natural host. This study aims to reassess and harmonize the phylogenetic placement and binomen of T. annae within the order Piroplasmida. Methods Four molecular phylogenetic trees were constructed using a maximum likelihood algorithm based on DNA alignments of: (i) near-complete 18S rRNA gene sequences (n = 76 and n = 93), (ii) near-complete and incomplete 18S rRNA gene sequences (n = 92), and (iii) tubulin-beta gene sequences (n = 32) from B. microti and B. microti -related parasites including those detected in dogs and foxes. Results All phylogenetic trees demonstrate that T. annae and its synonyms are not Theileria parasites but are most closely related with B. microti . The phylogenetic tree based on the 18S rRNA gene forms two separate branches with high bootstrap value, of which one branch corresponds to Babesia species infecting rodents, humans, and macaques, while the other corresponds to species exclusively infecting carnivores. Within the carnivore group, T. annae and its synonyms from distant regions segregate into a single clade with a highly significant bootstrap value corroborating their separate species identity. Conclusion Phylogenetic analysis clearly shows that T. annae and its synonyms do not pertain to Theileria and can be clearly defined as a separate species. Based on the facts that T. annae and its synonyms have not been shown to have a leukocyte stage, as expected in Theileria , do not infect humans and rodents as B. microti , and cluster phylogenetically as a separate species, this study proposes to name this parasite Babesia vulpes sp. nov., after its natural host, the red fox V. vulpes .