Piroplasmids of the genera Babesia, Theileria, and Cytauxzoon are tick-transmitted parasites with a high impact on animals and humans. They have complex life cycles in their definitive arthropod and intermediate vertebrate hosts involving numerous processes, including invasion of, and egress from, host cells, parasite growth, transformation, and migration. Like other parasitic protozoa, piroplasmids are equipped with different types of protease to fulfill many of such essential processes. Blockade of some key proteases, using inhibitors or antibodies, hinders piroplasmid growth, highlighting their potential usefulness in drug therapies and vaccine development. A better understanding of the functional significance of these enzymes will contribute to the development of improved control measures for the devastating animal and human diseases caused by these pathogens.

• MeSH
• Babesia * genetika   MeSH
• babezióza * parazitologie   MeSH
• klíšťata * parazitologie   MeSH
• lidé MeSH
• Piroplasmida * MeSH
• proteasy MeSH
• Theileria * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

BACKGROUND: The range of the ornate dog tick Dermacentor reticulatus is rapidly expanding in Europe. This tick species is the vector of canine babesiosis, caused by Babesia canis, and also plays a role in the transmission of Theileria equi and Babesia caballi in equids. METHODS: The geographic range of D. reticulatus in the Czech Republic was re-assessed, and an up-to-date distribution map is presented based on material and data obtained during a nationwide citizen science campaign. Received and flagged individuals of D. reticulatus were also analysed for the presence of B. canis DNA. RESULTS: In striking contrast to historical records, D. reticulatus was found in all regions of the Czech Republic, with most reports coming from the southeast and northwest of the country. Between February 2018 and June 2021, the project team received 558 photo reports of ticks and 250 packages containing ticks. Of the former, 71.1% were identified as Dermacentor sp. with the remainder identified as Ixodes sp., Haemaphysalis sp., Argas sp. or Hyalomma sp. The majority of specimens in the subset of ticks that were received (N = 610) were D. reticulatus (N = 568, 93.7%), followed by Ixodes ricinus and Hyalomma spp. A total of 783 adult D. reticulatus, either received (568) or collected by flagging (215), were tested for the presence of B. canis DNA using species-specific nested PCR targeting part of the 18S rRNA gene; B. canis DNA was demonstrated in 22 samples (2.81%). CONCLUSIONS: The continuous spread of D. reticulatus in the Czech Republic was documented in this study. In addition, DNA of B. canis was also detected in a number of ticks, suggesting the establishment of B. canis in the Czech Republic. These results suggest that veterinarians need to consider the possibility of canine babesiosis even in dogs without a history of travel.

• MeSH
• Babesia * genetika   MeSH
• babezióza * epidemiologie   MeSH
• Dermacentor * MeSH
• infestace klíšťaty * epidemiologie   veterinární   MeSH
• Ixodidae * MeSH
• klíště * MeSH
• nemoci psů * epidemiologie   MeSH
• občanská věda * MeSH
• psi MeSH
• Rhipicephalus sanguineus * MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

The Gram-negative, obligate intracellular tick-transmitted pathogen Anaplasma phagocytophilum can cause acute febrile diseases in humans and domestic animals. The expansion of the tick Ixodes ricinus (Linnaeus, 1758) in northern Europe due to climate change is of serious concern for animal and human health. The aim of the present study was to investigate the impact of A. phagocytophilum infection in moose Alces alces (Linnaeus) calves by evaluating the carcass weights of infected and non-infected animals and examining animal tissues samples for co-infections with either species of Babesia Starcovici, 1893 or bacteria of the genus Bartonella. The carcasses of 68 free-ranging moose calves were weighed by hunters during the hunting seasons from 2014 to 2017 in two regions in southern Norway and spleen samples were collected. Anaplasma phagocytophilum was detected in moose sampled from locations infected with ticks with a prevalence of 82% (n = 46). The carcass weights of A. phagocytophilum-infected calves (n = 46) and non-infected (n = 22) calves were compared. Although the average weight of infected calves (45.6 kg) was lower than that of non-infected calves (46.5 kg), the difference was not statistically significant. Three different variants of the bacterium 16S rRNA gene were identified. The average weight of animals infected with variant I was 49.9 kg, whereas that of animals infected with variant III was 42.0 kg, but the difference was not statistically significant (p = 0.077). Co-infections of A. phagocytophilum with Bartonella spp. or with Babesia spp. were found in 20 and two calves, respectively. A triple infection was found in two calves. Sequence analysis of the 18S rRNA gene of Babesia-positive samples revealed the presence of Babesia cf. odocoilei (Emerson et Wright, 1970). Strains of Bartonella closely related to Bartonella bovis (Bermond, Boulouis, Heller, Laere, Monteil, Chomel, Sander, Dehio et Piemont, 2002) were identified based on phylogenetic analysis of the gltA and rpoB genes. The loss of body mass in moose calves in the tick-infected site was probably influenced by multiple factors.

• MeSH
• Anaplasma phagocytophilum * klasifikace   genetika   izolace a purifikace   MeSH
• Babesia genetika   MeSH
• Bartonella genetika   MeSH
• DNA bakterií chemie   genetika   izolace a purifikace   MeSH
• ehrlichióza komplikace   epidemiologie   patologie   veterinární   MeSH
• fylogeneze MeSH
• oligonukleotidy chemie   MeSH
• polymerázová řetězová reakce veterinární   MeSH
• sekvence nukleotidů MeSH
• slezina mikrobiologie   patologie   MeSH
• tělesná hmotnost MeSH
• vysoká zvěř * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Norsko MeSH

We developed a PCR assay for the detection of Babesia odocoilei based on the 18S rRNA gene. Multiple specimens of B. odocoilei were examined, and the assay consistently produced a small specific PCR product of 306 bp. The PCR assay was also challenged with DNA from 13 other Babesia species and 2 Theileria species, originating from 10 different host species; however, nonspecific DNA amplification and multiple banding patterns were observed, and the amplicon banding patterns varied between different isolates of the same species. Sensitivity was determined to be 6.4 pg of DNA, and an estimated 0.0001% parasitism. This assay can be utilized for species-specific differential detection of B. odocoilei.

• MeSH
• Babesia * genetika   MeSH
• babezióza * diagnóza   MeSH
• polymerázová řetězová reakce veterinární   MeSH
• protozoální DNA genetika   MeSH
• RNA ribozomální 18S genetika   MeSH
• Theileria * genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Theileria equi and Babesia caballi are protozoan agents causing equine piroplasmosis, endemic in countries all over the world. The aim of this study was to detect antibodies to T. equi and B. caballi in horses in the Czech Republic and to investigate the origin of the infection. Blood sera from 711 horses were examined with competitive enzyme-linked immunosorbent assay; positive samples were verified with indirect fluorescence immunoassay. Antibodies to T. equi and B. caballi were detected in eight (1.1%) and three (0.4%) horses, respectively. Infection with T. equi was confirmed by PCR and sequencing in the blood of five serologically positive horses. An autochthonous origin of T. equi infection could not be excluded in two (0.3%) horses. Intensive movement of horses across European countries and the expanding occurrence of competent tick vector Dermacentor reticulatus in the Czech Republic create an increasing risk of establishing active foci of equine piroplasmosis in the country.

• MeSH
• Babesia * genetika   MeSH
• babezióza * epidemiologie   MeSH
• koně MeSH
• nemoci koní * epidemiologie   MeSH
• skot MeSH
• Theileria * genetika   MeSH
• theilerióza * epidemiologie   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Free ranging ungulates, represented in Europe mostly by several deer species, are important hosts for ticks and reservoirs of tick-borne infections. A number of studies have focused on the prevalence of tick borne pathogens in deer chiefly with the aim to determine their potential role as reservoir hosts for important human and livestock pathogens. However, genetic similarity of Babesia spp. forming a group commonly termed as a clade VI that accommodates the deer piroplasms, complicates this task and has led to the description of a bewildering array of poorly characterised strains. This study aims to resolve this issue by using two independent genetic loci, nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit I genes, used in parallel to identify Babesia isolates in free-ranging red, sika, and roe deer in two areas of their co-occurrence in the Czech Republic. The COX1 loci, in contrast to 18S rRNA gene, shows a clear difference between interspecific and intraspecific variation at the nucleotide level. The findings confirm B. divergens, Babesia sp. EU1 and B. capreoli in studied deer species as well as common presence of another unnamed species that matches a taxon previously referred to as Babesia sp. or Babesia cf. odocoilei or Babesia CH1 group in several other sites throughout Europe. The invasive sika deers enter the life cycle of at least three piroplasmid species detected in native deer fauna. The presence of B. divergens in both sika and red deer in an area where bovine babesiosis is apparently absent raises important questions regarding the epidemiology, host specificity and taxonomic status of the parasite.

• MeSH
• Babesia klasifikace   genetika   MeSH
• babezióza parazitologie   virologie   MeSH
• cyklooxygenasa 1 genetika   MeSH
• fylogeneze MeSH
• molekulární evoluce MeSH
• protozoální DNA genetika   MeSH
• protozoální proteiny genetika   MeSH
• ribozomální DNA genetika   MeSH
• RNA ribozomální 18S genetika   MeSH
• sekvenční analýza DNA metody   MeSH
• vysoká zvěř parazitologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

BACKGROUND: Babesia spp. are apicomplexan parasites which infect a wide range of mammalian hosts. Historically, most Babesia species were described based on the assumed host specificity and morphological features of the intraerythrocytic stages. New DNA-based approaches challenge the traditional species concept and host specificity in Babesia. Using such tools, the presence of Babesia DNA was reported in non-specific mammalian hosts, including B. canis in feces and tissues of insectivorous bats, opening questions on alternative transmission routes. The aim of the present study was to evaluate if B. canis DNA can be detected in tissues of laboratory rodents following oral inoculation with infected ticks. METHODS: Seventy-five questing adult Dermacentor reticulatus ticks were longitudinally cut in two halves and pooled. Each pool consisted of halves of 5 ticks, resulting in two analogous sets. One pool set (n = 15) served for DNA extraction, while the other set (n = 15) was used for oral inoculation of experimental animals (Mus musculus, line CD-1 and Meriones unguiculatus). Blood was collected three times during the experiment (before the inoculation, at 14 days post-inoculation and at 30 days post-inoculation). All animals were euthanized 30 days post-inoculation. At necropsy, half of the heart, lung, liver, spleen and kidneys were collected from each animal. The presence of Babesia DNA targeting the 18S rRNA gene was evaluated from blood and tissues samples. For histopathology, the other halves of the tissues were used. Stained blood smears were used for the light microscopy detection of Babesia. RESULTS: From the 15 pools of D. reticulatus used for the oral inoculation, six were PCR-positive for B. canis. DNA of B. canis was detected in blood and tissues of 33.3% of the animals (4 out of 12) inoculated with a B. canis-positive pool. No Babesia DNA was detected in the other 18 animals which received B. canis-negative tick pools. No Babesia was detected during the histological examination and all blood smears were microscopically negative. CONCLUSIONS: Our findings demonstrate that B. canis DNA can be detected in tissues of mammalian hosts following ingestion of infected ticks and opens the question of alternative transmission routes for piroplasms.

• MeSH
• aplikace orální MeSH
• Babesia genetika   MeSH
• babezióza krev   parazitologie   MeSH
• Dermacentor parazitologie   MeSH
• Gerbillinae MeSH
• hlodavci parazitologie   MeSH
• infestace klíšťaty parazitologie   MeSH
• myši MeSH
• protozoální DNA analýza   MeSH
• RNA ribozomální 18S genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

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.

• MeSH
• Babesia klasifikace   genetika   růst a vývoj   MeSH
• babezióza parazitologie   MeSH
• fylogeneze * MeSH
• lidé MeSH
• Piroplasmida klasifikace   genetika   růst a vývoj   MeSH
• protozoální geny genetika   MeSH
• stadia vývoje genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Canine babesiosis caused by Babesia canis sensu stricto became an emerging disease of dogs across Europe calling for attention also in countries where it was an only rare imported disease. An easy accessibility of molecular methods and the growing amount of sequencing data led to the description of intraspecific variability in 18S rDNA sequences designated as "genotypes". Using material from a homogenous cohort of dogs with microscopically confirmed canine babesiosis caused by B. canis, we evaluated Babesia intraspecific variability and amplification sensitivity of three different genes (18S rDNA, COI, Cytb) to assess their potential as diagnostic or phylogenetic markers. In raw sequencing data obtained, we observed at least 3 ambiguous positions in up to 86% of chromatograms within the ∼560 bp fragment of 18S rDNA suggesting the existence of several, not identical copies of this gene. Our COI haplotype analysis resulted in a star-like pattern indicating a recent origin of most haplotypes, but not supporting the existence of two dominant haplotypes. Similarly, the Cytb sequences obtained from samples with all variants of 18S rDNA were identical. We corroborate previous observations from three other European countries and bring the evidence of the existence of 18S rDNA paralogs in B. canis genome replacing currently used "genotype" theory.

• MeSH
• Babesia genetika   MeSH
• babezióza krev   diagnóza   parazitologie   MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• genetické markery MeSH
• genom protozoální MeSH
• genotyp * MeSH
• haplotypy MeSH
• kohortové studie MeSH
• mitochondrie genetika   MeSH
• nemoci psů diagnóza   parazitologie   MeSH
• protozoální DNA genetika   MeSH
• psi MeSH
• RNA ribozomální 18S genetika   MeSH
• sekvenční analýza DNA MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

Babesiosis is a tick-transmitted zoonosis caused by apicomplexan parasites of the genus Babesia. Treatment of this emerging malaria-related disease has relied on antimalarial drugs and antibiotics. The proteasome of Plasmodium, the causative agent of malaria, has recently been validated as a target for anti-malarial drug development and therefore, in this study, we investigated the effect of epoxyketone (carfilzomib, ONX-0914 and epoxomicin) and boronic acid (bortezomib and ixazomib) proteasome inhibitors on the growth and survival of Babesia. Testing the compounds against Babesia divergens ex vivo revealed suppressive effects on parasite growth with activity that was higher than the cytotoxic effects on a non-transformed mouse macrophage cell line. Furthermore, we showed that the most-effective compound, carfilzomib, significantly reduces parasite multiplication in a Babesia microti infected mouse model without noticeable adverse effects. In addition, treatment with carfilzomib lead to an ex vivo and in vivo decrease in proteasome activity and accumulation of polyubiquitinated proteins compared to untreated control. Overall, our results demonstrate that the Babesia proteasome is a valid target for drug development and warrants the design of potent and selective B. divergens proteasome inhibitors for the treatment of babesiosis.

• MeSH
• Babesia microti účinky léků   genetika   růst a vývoj   MeSH
• Babesia účinky léků   genetika   růst a vývoj   MeSH
• babezióza farmakoterapie   MeSH
• buněčné linie MeSH
• inhibitory proteasomu aplikace a dávkování   škodlivé účinky   farmakologie   terapeutické užití   MeSH
• kyseliny boronové farmakologie   MeSH
• makrofágy účinky léků   parazitologie   MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• oligopeptidy farmakologie   MeSH
• proteasomový endopeptidasový komplex účinky léků   MeSH
• proteom účinky léků   genetika   MeSH
• systémy cílené aplikace léků * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• validační studie MeSH