Ticks are important vectors of various microorganisms, including bacteria. In this study, we examined Hyalomma aegyptium ticks collected from 240 spur-thighed tortoises Testudo graeca at 42 localities in the Mediterranean and Middle East and analysed them for the presence of bacteria of the genera Anaplasma, Borrelia, Coxiella, and Rickettsia. Altogether, 576 out of 928 analysed ticks (62.1%) were positive for at least one of the tested bacteria. The highest prevalence in individual ticks was found for Borrelia turcica (43.6%), followed by Rickettsia (12.3%) and Anaplasma (6.1%). No sample was positive for Coxiella burnetii. Among Rickettsia, we detected two species, Rickettsia africae and Rickettsia aeschlimannii, and also other unspecified Rickettsia. Anaplasma (100% identity with A. phagocytophilum) was detected at 15 (35%) out of 42 studied localities, any of Rickettsia at 28 (67%), and B. turcica at 32 (76%) localities. The geographic distribution of the studied microorganisms varied, with none of them detected in Syria, and only Rickettsia spp. detected in Morocco. Sequence analysis revealed substantial genetic variability in all detected agents, with the most variable (36 new haplotypes) being glpQ gene used as a marker for B. turcica. We also analysed the prevalence of various co-infections among studied ticks, with the mean number of co-infected ticks per tortoise increased with the number of ticks per tortoise. However, the frequencies of co-infected ticks do not indicate the presence of antagonistic or synergistic facilitative interactions between the agents. According to our data, we could expect that the eco-epidemiological importance of H. aegyptium does not stem from their tortoise hosts but rather from the low host specificity of its larvae and nymphs, feeding on a wider spectrum of reptilian, avian, and mammalian hosts.
- MeSH
- Anaplasma * isolation & purification MeSH
- Borrelia isolation & purification MeSH
- Coxiella isolation & purification genetics MeSH
- Tick Infestations veterinary epidemiology parasitology MeSH
- Ixodidae * microbiology growth & development MeSH
- Rickettsia * isolation & purification MeSH
- Turtles * microbiology MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Middle East MeSH
- Mediterranean Region MeSH
BACKGROUND: Ticks, hematophagous Acari, pose a significant threat by transmitting various pathogens to their vertebrate hosts during feeding. Despite advances in tick genomics, high-quality genomes were lacking until recently, particularly in the genus Ixodes, which includes the main vectors of Lyme disease. RESULTS: Here, we present the genome sequences of four tick species, derived from a single female individual, with a particular focus on the European species Ixodes ricinus, achieving a chromosome-level assembly. Additionally, draft assemblies were generated for the three other Ixodes species, I. persulcatus, I. pacificus, and I. hexagonus. The quality of the four genomes and extensive annotation of several important gene families have allowed us to study the evolution of gene repertoires at the level of the genus Ixodes and of the tick group. We have determined gene families that have undergone major amplifications during the evolution of ticks, while an expression atlas obtained for I. ricinus reveals striking patterns of specialization both between and within gene families. Notably, several gene family amplifications are associated with a proliferation of single-exon genes-most strikingly for fatty acid elongases and sulfotransferases. CONCLUSIONS: The integration of our data with existing genomes establishes a solid framework for the study of gene evolution, improving our understanding of tick biology. In addition, our work lays the foundations for applied research and innovative control targeting these organisms.
Tularemie je zoonóza, jejíž původcem je Francisella tularensis, gramnegativní aerobní bakterie patřící do třídy Gammaproteobacteria a čeledi Francisellaceae. Navzdory skutečnosti, že F. tularensis může mít významný dopad na zdraví člověka, existuje o aktuálním výskytu tohoto patogenu v různých hematofágních členovcích pouze velmi málo údajů. Cílem studie bylo provést rozsáhlý molekulární screening různých potenciálních hematofágních vektorů: klíšťat (4348 jedinců druhů Ixodes ricinus, Dermacentor reticulatus a Haemaphysalis concinna), komárů (4100 jedinců druhu Aedes vexans) a muchniček (6900 jedinců Simulium spp.) na přítomnost F. tularensis na Břeclavsku v roce 2022. Pozitivní byly pouze 2 vzorky, které obsahovaly DNA specifickou pro F. tularensis subsp. holarctica. Oba vzorky pocházely z klíštěte D. reticulatus a to jak po infestaci srnce, tak jednoho směsného vzorku nasbíraných klíšťat (n = 10). Oba pozitivní vzorky byly sekvenovány a byla potvrzena přítomnost F. tularensis subsp. holarctica. Přítomnost
Tularemia is a zoonosis caused by Francisella tularensis, a gram-negative aerobic bacterium belonging to the class of Gammaproteobacteria and the family Francisellaceae. Despite its undeniable importance for human health, there is little data on the current distribution of F. tularensis in various hematophagous arthropods. The aim of this study was to perform a mass molecular screening of different possible hematophagous vectors: ticks (4348 ticks of the species Ixodes ricinus, Dermacentor reticulatus, and Haemaphysalis concinna), mosquitoes (4100 specimens of Aedes vexans), and blackflies (6900 specimens of the Simulium spp.) for the presence of F. tularensis in the Břeclav district in 2022. Only two specimens were positive for the specific DNA of Francisella tularensis subsp. holarctica. Both samples originated from D. reticulatus, one collected from infested roe deer and the other included in a pooled sample (n = 10). Both positive samples were sequenced, and the presence of F. tularensis subsp. holarctica was confirmed. In addition, the absence of F. tularensis in mosquitoes and black flies was documented.
- MeSH
- Francisella tularensis * pathogenicity MeSH
- Genetic Techniques MeSH
- Disease Vectors * MeSH
- Mosquito Vectors microbiology MeSH
- Communicable Disease Control * methods MeSH
- Humans MeSH
- Tick-Borne Diseases epidemiology microbiology transmission MeSH
- Simuliidae microbiology MeSH
- Tularemia epidemiology microbiology transmission MeSH
- Disease Reservoirs microbiology MeSH
- Check Tag
- Humans MeSH
- Geographicals
- Czech Republic MeSH
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- MeSH
- Disease Notification MeSH
- Communicable Diseases, Imported epidemiology MeSH
- Disease Vectors * MeSH
- Humans MeSH
- Tick-Borne Diseases MeSH
- Mosquito-Borne Diseases epidemiology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Tables MeSH
- Geographicals
- Czech Republic MeSH
Viral infection may represent a stress condition to the host cell. Cells react to it by triggering the defence programme to restore homeostasis and these events may in turn impact the viral replication. The knowledge about tick-borne encephalitis virus (TBEV) infection-associated stress is limited. Here we investigated the interplay between TBEV infection and stress pathways in PMJ2-R mouse macrophage cell line, as macrophages are the target cells in early phases of TBEV infection. First, to determine how stress influences TBEV replication, the effect of stress inducers H2O2 and tunicamycin (TM) was tested. Viral multiplication was decreased in the presence of both stress inducers suggesting that the stress and cellular stress responses restrict the virus replication. Second, we investigated the induction of oxidative stress and endoplasmic reticulum (ER) stress upon TBEV infection. The level of oxidative stress was interrogated by measuring the reactive oxygen species (ROS). ROS were intermittently increased in infected cells at 12 hpi and at 72 hpi. As mitochondrial dysfunction may result in increased ROS level, we evaluated the mitochondrial homeostasis by measuring the mitochondrial membrane potential (MMP) and found that TBEV infection induced the hyperpolarization of MMP. Moreover, a transient increase of gene expression of stress-induced antioxidative enzymes, like p62, Gclm and Hmox1, was detected. Next, we evaluated the ER stress upon TBEV infection by analysing unfolded protein responses (UPR). We found that infection induced gene expression of two general sensors BiP and CHOP and activated the IRE1 pathway of UPR. Finally, since the natural transmission route of TBEV from its tick vector to the host is mediated via tick saliva, the impact of tick saliva from Ixodes ricinus on stress pathways in TBEV-infected cells was tested. We observed only marginal potentiation of UPR pathway. In conclusion, we found that TBEV infection of PMJ2-R cells elicits the changes in redox balance and triggers cellular stress defences, including antioxidant responses and the IRE1 pathway of UPR. Importantly, our results revealed the negative effect of stress-evoked events on TBEV replication and only marginal impact of tick saliva on stress cellular pathways.
- MeSH
- Cell Line MeSH
- Encephalitis, Tick-Borne * MeSH
- Mice MeSH
- Hydrogen Peroxide metabolism MeSH
- Protein Serine-Threonine Kinases metabolism MeSH
- Reactive Oxygen Species metabolism MeSH
- Virus Replication MeSH
- Encephalitis Viruses, Tick-Borne * genetics MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Relapsing fever (RF), a vector-borne disease caused by Borrelia spp., is characterized by recurring febrile episodes due to repeated bouts of bacteremia. RF spirochetes can be geographically and phylogenetically divided into two distinct groups; Old World RF Borrelia (found in Africa, Asia, and Europe) and New World RF Borrelia (found in the Americas). While RF is a rarely reported disease in the Americas, RF is prevalent in endemic parts of Africa. Despite phylogenetic differences between Old World and New World RF Borrelia and higher incidence of disease associated with Old World RF spirochete infection, genetic manipulation has only been described in New World RF bacteria. Herein, we report the generation of genetic tools for use in the Old World RF spirochete, Borrelia duttonii. We describe methods for transformation and establish shuttle vector- and integration-based approaches for genetic complementation, creating green fluorescent protein (gfp)-expressing B. duttonii strains as a proof of principle. Allelic exchange mutagenesis was also used to inactivate a homolog of the Borrelia burgdorferi p66 gene, which encodes an important virulence factor, in B. duttonii and demonstrate that this mutant was attenuated in a murine model of RF. Finally, the B. duttonii p66 mutant was complemented using shuttle vector- and cis integration-based approaches. As expected, complemented p66 mutant strains were fully infectious, confirming that P66 is required for optimal mammalian infection. The genetic tools and techniques reported herein represent an important advancement in the study of RF Borrelia that allows for future characterization of virulence determinants and colonization factors important for the enzootic cycle of Old World RF spirochetes.
- MeSH
- Borrelia * genetics classification MeSH
- Humans MeSH
- Mice MeSH
- Relapsing Fever * microbiology MeSH
- Genetic Complementation Test MeSH
- Green Fluorescent Proteins genetics MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Background: Tick-borne encephalitis (TBE) is caused by the tick-borne encephalitis virus (TBEV). TBEV infection can cause symptoms of central nervous system (CNS) inflammation and result in severe consequences including death. TBE is an increasing health threat in the Czech Republic and elsewhere in Europe. In 2020, 23% of 3734 TBE cases reported to the European Centre for Disease Prevention and Control were from the Czech Republic. TBE vaccination is universally recommended in the Czech Republic, but a full analysis of TBE vaccine effectiveness (VE) in the Czech Republic has not been published. Methods: TBE is a notifiable disease in the Czech Republic with mandatory reporting of cases (i.e., laboratory-confirmed TBEV infected patient with symptoms of CNS inflammation) and vaccination history to public health authorities. TBE VE was estimated using the screening method utilizing public health surveillance data from 2018 to 2022 and online household surveys of the general population on TBE vaccine uptake conducted in 2019-2022. Results: In 2018-2022, 3648 TBE cases were reported in the Czech Republic; 98.1% (3105/3166) of TBE cases with known vaccination history were unvaccinated. Among 42,671 persons surveyed from the general population who had known TBE vaccination history, 66.5% were unvaccinated. VE against TBE was 97.6% (95% confidence interval 95.7-98.7). When stratified by age group, VE was 97.1% (88.4-99.3) in 1-15 years of age, 97.9% (95.3-99.0) in 16-59 years of age, and 96.9% (90.5-99.0) in ≥60 years of age. TBE vaccination averted an estimated 1020 TBE cases in the Czech Republic from 2018 to 2022. Conclusions: This first published study with a full analysis of TBE VE in the Czech Republic showed that vaccination was highly effective for the prevention of TBE including in children, an age group with increasing TBE disease burden. Vaccination averted hundreds of TBE cases and hospitalizations despite the relatively low compliance with TBE vaccine recommendations. To prevent additional TBE cases in the Czech Republic, enhanced efforts to increase TBE vaccine uptake are needed.
- MeSH
- Child MeSH
- Adult MeSH
- Encephalitis, Tick-Borne * prevention & control epidemiology MeSH
- Infant MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Child, Preschool MeSH
- Aged MeSH
- Vaccination * MeSH
- Viral Vaccines * administration & dosage MeSH
- Encephalitis Viruses, Tick-Borne * immunology MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Infant MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Child, Preschool MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
The brown dog tick, Rhipicephalus sanguineus is a complex of tick species with an unsettled species concept. In Europe, R. sanguineus is considered mainly a Mediterranean tick with sporadic findings in central and northern Europe. R. sanguineus is known as a vector of a range of pathogens of medical and veterinary importance, most of which not yet reported as autochthonous in Hungary. A total of 1839 ticks collected by veterinarians from dogs and cats were obtained in Hungary. The study aims at precise determination of ticks identified as R. sanguineus and detection of pathogens in collected ticks. All ticks were morphologically determined and 169 individuals were identified as R. sanguineus. A subset of 15 ticks was selected for molecular analysis (16S rDNA, 12S rDNA, COI). Phylogenetic analyses invariably placed sequences of all three markers into a single haplotype identified as R. sanguineus sensu stricto. All 169 brown dog ticks were tested for the presence of A. platys, E. canis, R. conorii, B. vogeli and H. canis. None of the investigated ticks was positive for the screened pathogens, though A. phagocytophilum sequence was detected in a single tick.
- MeSH
- Anaplasma * MeSH
- Ehrlichia canis isolation & purification genetics MeSH
- Phylogeny * MeSH
- Tick Infestations * veterinary parasitology MeSH
- Cats parasitology MeSH
- Tick-Borne Diseases veterinary microbiology parasitology MeSH
- Dog Diseases * parasitology diagnosis MeSH
- Dogs MeSH
- Rhipicephalus sanguineus * microbiology MeSH
- Rickettsia conorii isolation & purification genetics MeSH
- RNA, Ribosomal, 16S analysis genetics MeSH
- RNA, Ribosomal * MeSH
- Animals MeSH
- Check Tag
- Cats parasitology MeSH
- Male MeSH
- Dogs MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Case Reports MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Hungary MeSH
The emergence of tick-borne encephalitis (TBE) in Europe marked several significant milestones. The discovery of TBE in Czechoslovakia in 1948, with Gallia and Krejčí simultaneously isolating the TBE virus (TBEV) from human samples for the first time in Europe outside the Soviet Union, was pivotal. Subsequent TBEV isolation from ticks suggested the viral transmission via this vector. In 1951, the outbreak in Rožňava in Slovakia (Czechoslovakia) revealed an unexpected mode of transmission, unpasteurized milk from a local dairy, challenging existing understanding. Investigations exposed illicit practices of mixing cow's milk with goat's milk for economic gains. Laboratory research confirmed the outbreak was caused by TBEV, which was substantiated by serological analyses. This was the first and largest documented alimentary TBE outbreak in history. In this review, we delve into both published sources and unpublished archival data, offering a comprehensive understanding of these historic accomplishments and shedding light on these pivotal moments.