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"Rudenko, Natalie"
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Metamorphoses of Lyme disease spirochetes: phenomenon of Borrelia persisters
by
Rudenko, Natalie
,
Golovchenko, Maryna
,
Kybicova, Katerina
in
Acidity
,
Anopheles
,
Anti-Bacterial Agents - pharmacology
2019
The survival of spirochetes from the
Borrelia burgdorferi
(
sensu lato
) complex in a hostile environment is achieved by the regulation of differential gene expression in response to changes in temperature, salts, nutrient content, acidity fluctuation, multiple host or vector dependent factors, and leads to the formation of dormant subpopulations of cells. From the other side, alterations in the level of gene expression in response to antibiotic pressure leads to the establishment of a persisters subpopulation. Both subpopulations represent the cells in different physiological states. “Dormancy” and “persistence” do share some similarities, e.g. both represent cells with low metabolic activity that can exist for extended periods without replication, both constitute populations with different gene expression profiles and both differ significantly from replicating forms of spirochetes. Persisters are elusive, present in low numbers, morphologically heterogeneous, multi-drug-tolerant cells that can change with the environment. The definition of “persisters” substituted the originally-used term “survivors”, referring to the small bacterial population of
Staphylococcus
that survived killing by penicillin. The phenomenon of persisters is present in almost all bacterial species; however, the reasons why
Borrelia
persisters form are poorly understood. Persisters can adopt varying sizes and shapes, changing from well-known forms to altered morphologies. They are capable of forming round bodies, L-form bacteria, microcolonies or biofilms-like aggregates, which remarkably change the response of
Borrelia
to hostile environments. Persisters remain viable despite aggressive antibiotic challenge and are able to reversibly convert into motile forms in a favorable growth environment. Persisters are present in significant numbers in biofilms, which has led to the explanation of biofilm tolerance to antibiotics. Considering that biofilms are associated with numerous chronic diseases through their resilient presence in the human body, it is not surprising that interest in persisting cells has consequently accelerated. Certain diseases caused by pathogenic bacteria (e.g. tuberculosis, syphilis or leprosy) are commonly chronic in nature and often recur despite antibiotic treatment. Three decades of basic and clinical research have not yet provided a definite answer to the question: is there a connection between persisting spirochetes and recurrence of Lyme disease in patients?
Journal Article
Sexual Transmission of Lyme Borreliosis? The Question That Calls for an Answer
2021
Transmission of the causative agents of numerous infectious diseases might be potentially conducted by various routes if this is supported by the genetics of the pathogen. Various transmission modes occur in related pathogens, reflecting a complex process that is specific for each particular host–pathogen system that relies on and is affected by pathogen and host genetics and ecology, ensuring the epidemiological spread of the pathogen. The recent dramatic rise in diagnosed cases of Lyme borreliosis might be due to several factors: the shifting of the distributional range of tick vectors caused by climate change; dispersal of infected ticks due to host animal migration; recent urbanization; an increasing overlap of humans’ habitat with wildlife reservoirs and the environment of tick vectors of Borrelia; improvements in disease diagnosis; or establishment of adequate surveillance. The involvement of other bloodsucking arthropod vectors and/or other routes of transmission (human-to-human) of the causative agent of Lyme borreliosis, the spirochetes from the Borrelia burgdorferi sensu lato complex, has been speculated to be contributing to increased disease burden. It does not matter how controversial the idea of vector-free spirochete transmission might seem in the beginning. As long as evidence of sexual transmission of Borrelia burgdorferi both between vertebrate hosts and between tick vectors exists, this question must be addressed. In order to confirm or refute the existence of this phenomenon, which could have important implications for Lyme borreliosis epidemiology, the need of extensive research is obvious and required.
Journal Article
Genomic Confirmation of Borrelia garinii , United States
2023
Lyme disease is a multisystem disorder primarily caused by Borrelia burgdorferi sensu lato. However, B. garinii, which has been identified on islands off the coast of Newfoundland and Labrador, Canada, is a cause of Lyme disease in Eurasia. We report isolation and whole-genome nucleotide sequencing of a B. garinii isolate from a cotton mouse (Peromyscus gossypinus) in South Carolina, USA. We identified a second B. garinii isolate from the same repository. Phylogenetic analysis does not associate these isolates with the previously described isolates of B. garinii from Canada.
Journal Article
A human secretome library screen reveals a role for Peptidoglycan Recognition Protein 1 in Lyme borreliosis
2020
Lyme disease, the most common vector-borne illness in North America, is caused by the spirochete Borrelia burgdorferi . Infection begins in the skin following a tick bite and can spread to the hearts, joints, nervous system, and other organs. Diverse host responses influence the level of B . burgdorferi infection in mice and humans. Using a systems biology approach, we examined potential molecular interactions between human extracellular and secreted proteins and B . burgdorferi . A yeast display library expressing 1031 human extracellular proteins was probed against 36 isolates of B . burgdorferi sensu lato . We found that human Peptidoglycan Recognition Protein 1 (PGLYRP1) interacted with the vast majority of B . burgdorferi isolates. In subsequent experiments, we demonstrated that recombinant PGLYRP1 interacts with purified B . burgdorferi peptidoglycan and exhibits borreliacidal activity, suggesting that vertebrate hosts may use PGLYRP1 to identify B . burgdorferi . We examined B . burgdorferi infection in mice lacking PGLYRP1 and observed an increased spirochete burden in the heart and joints, along with splenomegaly. Mice lacking PGLYRP1 also showed signs of immune dysregulation, including lower serum IgG levels and higher levels of IFNγ, CXCL9, and CXCL10.Taken together, our findings suggest that PGLYRP1 plays a role in the host’s response to B . burgdorferi and further demonstrate the utility of expansive yeast display screening in capturing biologically relevant interactions between spirochetes and their hosts.
Journal Article
Natural selection and recombination at host-interacting lipoprotein loci drive genome diversification of Lyme disease and related bacteria
2024
Lyme disease (also called Lyme borreliosis in Europe), a condition caused by spirochete bacteria of the genus Borrelia , transmitted by hard-bodied Ixodes ticks, is currently the most prevalent and rapidly expanding tick-borne disease in the United States and Europe. Borrelia interspecies and intraspecies genome comparisons of Lyme disease-related bacteria are essential to reconstruct their evolutionary origins, track epidemiological spread, identify molecular mechanisms of human pathogenicity, and design molecular and ecological approaches to disease prevention, diagnosis, and treatment. These Lyme disease-associated bacteria harbor complex genomes that encode many genes that do not have homologs in other organisms and are distributed across multiple linear and circular plasmids. The functional significance of most of the plasmid-borne genes and the multipartite genome organization itself remains unknown. Here we sequenced, assembled, and analyzed whole genomes of 47 Borrelia isolates from around the world, including multiple isolates of the human pathogenic species. Our analysis elucidates the evolutionary origins, historical migration, and sources of genomic variability of these clinically important pathogens. We have developed web-based software tools (BorreliaBase.org) to facilitate dissemination and continued comparative analysis of Borrelia genomes to identify determinants of human pathogenicity.
Journal Article
Hedgehogs, Squirrels, and Blackbirds as Sentinel Hosts for Active Surveillance of Borrelia miyamotoi and Borrelia burgdorferi Complex in Urban and Rural Environments
by
Černá Bolfíková, Barbora
,
Majerová, Karolina
,
Papežík, Petr
in
Animals
,
Arachnids
,
Biological materials
2020
Lyme borreliosis (LB), caused by spirochetes of the Borrelia burgdorferi sensu lato (s.l.) complex, is one of the most common vector-borne zoonotic diseases in Europe. Knowledge about the enzootic circulation of Borrelia pathogens between ticks and their vertebrate hosts is epidemiologically important and enables assessment of the health risk for the human population. In our project, we focused on the following vertebrate species: European hedgehog (Erinaceus europaeus), Northern white-breasted hedgehog (E. roumanicus), Eurasian red squirrel (Sciurus vulgaris), and Common blackbird (Turdus merula). The cadavers of accidentally killed animals used in this study constitute an available source of biological material, and we have confirmed its potential for wide monitoring of B. burgdorferi s.l. presence and genospecies diversity in the urban environment. High infection rates (90% for E. erinaceus, 73% for E. roumanicus, 91% for S. vulgaris, and 68% for T. merula) were observed in all four target host species; mixed infections by several genospecies were detected on the level of individuals, as well as in particular tissue samples. These findings show the usefulness of multiple tissue sampling as tool for revealing the occurrence of several genospecies within one animal and the risk of missing particular B. burgdorferi s.l. genospecies when looking in one organ alone.
Journal Article
Spatial and Temporal Variability in Prevalence Rates of Members of the Borrelia burgdorferi Species Complex in Ixodes ricinus Ticks in Urban, Agricultural and Sylvatic Habitats in Slovakia
2023
Lyme borreliosis (LB) is the most prevalent tick-borne human infection in Europe, with increasing incidence during the latest decades. Abundant populations of Ixodes ricinus, the main vector of the causative agent, spirochetes from the Borrelia burgdorferi sensu lato (Bbsl) complex, have been observed in urban and suburban areas of Europe, in general, and Slovakia, particularly. Understanding the spread of infectious diseases is crucial for implementing effective control measures. Global changes affect contact rates of humans and animals with Borrelia-infected ticks and increase the risk of contracting LB. The aim of this study was to investigate spatial and temporal variation in prevalence of Bbsl and diversity of its species in questing I. ricinus from three sites representing urban/suburban, natural and agricultural habitat types in Slovakia. Ixodes ricinus nymphs and adults were collected by dragging the vegetation in green areas of Bratislava town (urban/suburban habitat), in the Small Carpathians Mountains (natural habitat) (south-western Slovakia) and in an agricultural habitat at Rozhanovce in eastern Slovakia. Borrelia presence in ticks was detected by PCR and Bbsl species were identified by restriction fragment length polymorphism (RFLP). Borrelia burgdorferi s.l. species in coinfected ticks were identified by reverse line blot. Significant spatial and temporal variability in prevalence of infected ticks was revealed in the explored habitats. The lowest total prevalence was detected in the urban/suburban habitat, whereas higher prevalence was found in the natural and agricultural habitat. Six Bbsl species were detected by RFLP in each habitat type —B. burgdorferi sensu stricto (s.s.), B. afzelii, B. garinii, B. valaisiana, B. lusitaniae and B. spielmanii. Coinfections accounted for 3% of the total infections, whereby B. kurtenbachii was identified by RLB and sequencing in mixed infection with B. burgdorferi s.s, B. garinii and B. valaisiana. This finding represents the first record of B. kurtenbachii in questing I. ricinus in Slovakia and Europe. Variations in the proportion of Bbsl species were found between nymphs and adults, between years and between habitat types. Spatial variations in prevalence patterns and proportion of Bbsl species were also confirmed between locations within a relatively short distance in the urban habitat. Habitat-related and spatial variations in Borrelia prevalence and distribution of Bbsl species are probably associated with the local environmental conditions and vertebrate host spectrum. Due to the presence of Borrelia species pathogenic to humans, all explored sites can be ranked as areas with high epidemiological risk.
Journal Article
Advanced Molecular and Microscopic Diagnostics Suggest Congenital Borrelia Transmission: A Case Report
by
Rudenko, Natalie
,
Haque, Md. Hasibul
,
Golovchenko, Maryna
in
Antibiotics
,
Arachnids
,
Body fluids
2026
Lyme disease is by far the most common arthropod-borne disease in the Northern Hemisphere. It is caused by certain Borrelia species that are primarily transmitted to hosts by Ixodid ticks; however, transplacental transmission of the spirochete in both animals and humans has been reported. Here, we report imaging of intact spirochetes in an archived placental tissue sample that is immunoreactive to Borrelia antibodies and from which Borrelia DNA was isolated. Both mother and child showed evidence of seroreactivity to Borrelia spp. in the years following the child’s birth, although neither individual tested positive by the conventional two-tiered serological algorithm. Cultivation of viable spirochetes from a vaginal swab of the mother and from the urine of the child some years later supports the possibility of vector-free transmission of Borrelia from mother to child. By amplifying several genomic loci from the DNA of cultured and non-cultured Borrelia from blood and body fluid samples of the mother and child, the Borrelia in both were identified as the same species, Borrelia burgdorferi sensu stricto, a strain specific to North America.
Journal Article
First report of Usutu virus fatal infections in Chilean tinamous (Nothoprocta perdicaria), brahminy starlings (Sturnia pagodarum), and multiple other bird species in zoological gardens and wildlife in the Czech Republic
2026
Background
The Usutu virus (USUV;
Orthoflavivirus
,
Flaviviridae
) is a mosquito-borne pathogen causing fatal neuroinfections in susceptible wild and captive birds, particularly blackbirds, other passerines, and owls. Zoological gardens provide favourable conditions for the circulation of such viruses due to the proximity of diverse species and limited options for prevention.
Methods
Following the sudden death of several Chilean tinamous kept in the Brno zoological garden, we tested tissues sampled from 22 bird cadavers (from zoos, private owners, and free-living birds) for the presence of USUV and West Nile virus (WNV) RNA using duplex reverse transcription qPCR. Near-complete whole-genome sequences were acquired from positive samples by next-generation sequencing and subjected to phylogenetic analyses. Furthermore, serum samples from additional zoo animals and privately owned birds were screened for anti-flavivirus antibodies using ELISA and subsequently confirmed by the virus neutralization test.
Results
We report fatal USUV infections in multiple bird species from three zoological gardens in the Czech Republic. Duplex RT-qPCR targeting USUV and West Nile virus (WNV) detected USUV RNA in tissues from two Boreal owls (
Aegolius funereus
), one Eurasian pygmy owl (
Glaucidium passerinum
), two Brahminy starlings (
Sturnia pagodarum
), and four Chilean tinamous (
Nothoprocta perdicaria
). Additionally, three randomly found cadavers of free-living blackbirds (
Turdus merula
) tested positive. Pathological findings ranged from minimal pathological changes to pronounced hepatosplenomegaly with intestinal bleeding. Phylogenetic analysis of near-complete genome sequences assigned all viruses to the Europe 2 genetic lineage, revealing partial geographic clustering among isolates obtained in this study. Serological testing confirmed exposure in additional birds and demonstrated cross-neutralisation between anti-USUV and anti-WNV-positive sera.
Conclusions
In zoological gardens, flavivirus infections can cause substantial losses, even among rarely bred or endangered species. Given the zoonotic potential of both USUV and WNV, documenting their occurrence in avian hosts is important not only for animal health but also for human disease surveillance from the One Health perspective.
Journal Article
Role of Zoo-Housed Animals in the Ecology of Ticks and Tick-Borne Pathogens—A Review
2021
Ticks are ubiquitous ectoparasites, feeding on representatives of all classes of terrestrial vertebrates and transmitting numerous pathogens of high human and veterinary medical importance. Exotic animals kept in zoological gardens, ranches, wildlife parks or farms may play an important role in the ecology of ticks and tick-borne pathogens (TBPs), as they may serve as hosts for local tick species. Moreover, they can develop diseases of varying severity after being infected by TBPs, and theoretically, can thus serve as reservoirs, thereby further propagating TBPs in local ecosystems. The definite role of these animals in the tick–host-pathogen network remains poorly investigated. This review provides a summary of the information currently available regarding ticks and TBPs in connection to captive local and exotic wildlife, with an emphasis on zoo-housed species.
Journal Article