Rickettsiales Occurrence and Co-occurrence in Ixodes ricinus Ticks in Natural and Urban Areas.

Bacteria of Rickettsiaceae and Anaplasmataceae families include disease agents spread by Ixodes ricinus ticks, the most common tick vector in Europe. The aim of the study was to compare the prevalence and co-infection prevalence of particular tick-transmitted Rickettsiales members: Rickettsia spp. (further referred as Rs), Anaplasma phagocytophilum (Ap), and "Candidatus Neoehrlichia mikurensis" (CNM) in I. ricinus ticks in two types of areas, different in terms of human impact: natural and urban. Using additional data, we aimed at investigating co-occurrence of these Rickettsiales with Borreliella spp. A total of 4189 tick specimens, 2363 from the urban area (Warsaw park and forests) and 1826 from the natural area (forests and park in the vicinity of National Parks), were tested for the presence of Rickettsiales DNA by PCRs. The prevalence of selected Rickettsiales was twice higher in urban than natural areas (13.2% vs. 6.9%, respectively). In total ticks, the prevalence of Rs, Ap, and CNM was 6.5%, 5.3%, and 3.6% in urban areas vs. 4.4%, 1.1%, and 2.1% in natural areas, respectively. Co-infections of Rickettsiales were also more prevalent in urban areas (2.6% vs. 0.3%, respectively). The most common Rs was R. helvetica; also R. monacensis and novel "Candidatus Rickettsia mendelii" were detected. Positive association between Ap and CNM infections was discovered. Rickettsiales bacteria occurrence was not associated with Borreliella occurrence, but co-infections with these two groups were more common in ticks in urban areas. In conclusion, three groups of Rickettsiales constituted the important part of the tick pathogen community in Poland, especially in the urbanized central Poland (Mazovia). In the Warsaw agglomeration, there is a greater risk of encountering the I. ricinus tick infected with Rickettsiales and co-infected with Lyme spirochaetes, in comparison to natural areas. This finding raises the question whether cities might in fact be the hot spots for TBDs.

Long-term spatiotemporal stability and dynamic changes in helminth infracommunities of bank voles (Myodes glareolus) in NE Poland.

Parasites are considered to be an important selective force in host evolution but ecological studies of host-parasite systems are usually short-term providing only snap-shots of what may be dynamic systems. We have conducted four surveys of helminths of bank voles at three ecologically similar woodland sites in NE Poland, spaced over a period of 11 years, to assess the relative importance of temporal and spatial effects on helminth infracommunities. Some measures of infracommunity structure maintained relative stability: the rank order of prevalence and abundance of Heligmosomum mixtum, Heligmosomoides glareoli and Mastophorus muris changed little between the four surveys. Other measures changed markedly: dynamic changes were evident in Syphacia petrusewiczi which declined to local extinction, while the capillariid Aonchotheca annulosa first appeared in 2002 and then increased in prevalence and abundance over the remaining three surveys. Some species are therefore dynamic and both introductions and extinctions can be expected in ecological time. At higher taxonomic levels and for derived measures, year and host-age effects and their interactions with site are important. Our surveys emphasize that the site of capture is the major determinant of the species contributing to helminth community structure, providing some predictability in these systems.

Asymptomatic "Candidatus Neoehrlichia mikurensis" infections in immunocompetent humans.

In Europe, human infections with "Candidatus Neoehrlichia mikurensis" have mainly been restricted to immunocompromised patients. We report here the first cases of asymptomatic "Ca. Neoehrlichia mikurensis" infection in immunocompetent humans (5/316 [1.6%] were infected). Due to the potential threats of infections with "Ca. Neoehrlichia mikurensis" in healthy persons to the safety of the blood supply, further study of this phenomenon is required.

Long-term spatiotemporal stability and dynamic changes in the haemoparasite community of bank voles (Myodes glareolus) in NE Poland.

Long-term field studies on parasite communities are rare but provide a powerful insight into the ecological and evolutionary processes shaping host-parasite interactions. The aim of our study was to identify the principal factors regulating long-term trends in the haemoparasite communities of bank voles, and to this end, we sampled three semi-isolated populations of bank voles (n = 880) in 1999, 2002, 2006 and 2010 in the Mazury lake district region of NE Poland. Overall, 90.8 % of the bank voles harboured at least one of the species of haemoparasites studied. Whilst overall prevalence (all species combined) did not vary significantly between the surveys, different temporal changes were detected among voles in each of the three sites. In voles from Urwitalt, prevalence increased consistently with successive surveys, whereas in Talty, the peak years were 2002 and 2006, and in Pilchy, prevalence oscillated without a clear pattern. Across the study, bank voles harboured a mean of 1.75 ± 0.034 haemoparasite species, and species richness remained stable with no significant between-year fluctuations or trends. However, each of the five constituent species/genera showed a different pattern of spatio-temporal changes. The overall prevalence of Babesia microti was 4.9 %, but this varied significantly between years peaking in 2006 and declining again by 2010. For Bartonella spp., overall prevalence was 38.7 %, and this varied with year of study, but the temporal pattern of changes differed among the three sites. The overall prevalence of Haemobartonella (Mycoplasma) was 68.3 % with an increase in prevalence with year of study in all three sites. Hepatozoon erhardovae had an overall prevalence of 46.8 % but showed a marked reduction with each successive year of the study, and this was consistent in all three sites. The overall prevalence of Trypanosoma evotomys was 15.4 % varying significantly between sites, but showing temporal stability. While overall prevalence of all haemoparasites combined and species richness remained stable over the period of study, among the five haemoparasites, the pattern of spatiotemporal changes in prevalence and abundance of infections differed depending on parasite species. For some genera, host age was shown to play an important role, but a significant effect of host sex was found only for Haemobartonella spp.

Recombination, diversity and allele sharing of infectivity proteins between Bartonella species from rodents.

The alpha-Proteobacterium Bartonella is a common parasite of voles and mice, giving rise to short-lived (4 weeks to 2 months) infections. Here, we report high sequence diversity in genes of the VirB/VirD type IV secretion system (T4SS), amongst Bartonella from natural rodent populations in NE Poland. The VirB5 protein is predicted to consist of three conserved alpha helices separated by loops of variable length which include numerous indels. The C-terminal domain includes repeat stretches of KEK residues, reflecting underlying homopolymeric stretches of adenine residues. A total of 16 variants of VirB5, associated with host identity, but not bacterial taxon, were identified from 22 Bartonella isolates. One was clearly a recombinant from two others, another included an insertion of two KEK repeats. The virB5 gene appears to evolve via both mutation and recombination, as well as slippage mediated insertion/deletion events. The recombinational units are thought to be relatively short, as there was no evidence of linkage disequilibrium between virB5 and the bepA locus only 5.5 kb distant. The diversity of virB5 is assumed to be related to immunological role of this protein in Bartonella infections; diversity of virB5 may assist persistence of Bartonella in the rodent population, despite the relatively short (3-4 weeks) duration of individual infections. It is clear from the distribution of virB5 and bepA alleles that recombination within and between clades is widespread, and frequently crosses the boundaries of conventionally recognised Bartonella species.

Recombination within and between species of the alpha proteobacterium Bartonella infecting rodents.

Bartonella infections from wild mice and voles (Apodemus flavicollis, Mi. oeconomus, Microtus arvalis and Myodes glareolus) were sampled from a forest and old-field habitats of eastern Poland; a complex network of Bartonella isolates, referrable to B. taylorii, B. grahamii, B. birtlesii and B. doshiae, was identified by the sequencing of a gltA fragment, comparable to previous studies of Bartonella diversity in rodents. Nested clade analysis showed that isolates could be assigned to zero- and one-step clades which correlated with host identity and were probably the result of clonal expansion; however, sequencing of other housekeeping genes (rpoB, ribC, ftsZ, groEl) and the 16S RNA gene revealed a more complex situation with clear evidence of numerous recombinant events in which one or both Bartonella parents could be identified. Recombination within gltA was found to have generated two distinct variant clades, one a hybrid between B. taylorii and B. doshiae, the other between B. taylorii and B. grahamii. These recombinant events characterised the differences between the two-step and higher clades within the total nested cladogram, involved all four species of Bartonella identified in this work and appear to have played a dominant role in the evolution of Bartonella diversity. It is clear, therefore, that housekeeping gene phylogenies are not robust indicators of Bartonella diversity, especially when only a single gene (gltA or 16S RNA) is used. Bartonella clades infecting Microtus were most frequently involved in recombination and were most frequently tip clades within the cladogram. The role of Microtus in influencing the frequency of Bartonella recombination remains unknown.

Babesia spp. infections transmitted through blood transfusion.

Babesiosis in humans is caused by infection with various species of Babesia (Apicomplexa, Piroplasmida), mainly transmitted by an arthropod vector--Ixodes spp. ticks. This review will focus on blood transfusion as another mode of Babesia transmission, especially in endemic areas, as well as the impact of human babesiosis on transfusion medicine.

The effect of infectious agents on the prevalence of allergies.

PURPOSE: The role of infectious agents in allergy development is ambivalent. On one hand, there are reports of an association between a previous infection (especially a viral respiratory tract infection) and developing hypersensitivity to inhaled allergens, which in turn may increase the risk of developing allergic reactions. On the other hand, there are reports emphasizing a protective effect of a number of infectious agents against allergy development. The aim the study was to find possible associations between a past infectious or parasitic disease and an allergic condition. MATERIAL AND METHODS: The study population was a group of 18,648 subjects. The study, which was a part of the project: 'Implementation of a System for the Prevention and Early Detection of Allergic Diseases in Poland', was conducted in 9 selected regions of Poland and used the ECRHS and ISAAC questionnaires adapted for Europe. The following statistical tools were used: Pearson's chi-squared test, Fisher's exact test, and logistic regression. RESULTS: This research was an attempt to clear association between a history of measles or viral hepatitis and the likelihood of developing asthma, especially in males (χ2 = 5.29; p<0.05). Past parasitic disease showed a clear association with a suspected allergic rhinitis in various groups of patients (differing both in terms of sex and age). CONCLUSIONS: A history of some forms of either infectious or parasitic diseases has a measurable effect on the risk of developing allergies.

Effects of an MHC-DRB genotype and allele number on the load of gut parasites in the bank vole Myodes glareolus.

The major histocompatibility complex (MHC) genes code for the proteins responsible for pathogen recognition. The MHC class II DRB gene is multiplicated in the bank vole, Myodes glareolus, with different numbers of loci found in different individuals. Possessing large numbers of loci should increase the probability of pathogen recognition, but according to the optimality hypothesis, there is a cost of possessing too many MHC alleles. Using 454 technology, we determined the individual DRB allelic diversity and related it to the load of intestinal parasites in voles collected from three sites separated by a distance of 12 to 27 km. The analysis of six microsatellite loci revealed significant population structure (F(ST) = 0.07). The sites differed significantly in the prevalence and abundance of nematode species as well. We found two significant associations between MHC alleles and the intensity of the infection with the most prevalent nematode, Aspiculuris tetraptera. One of these associations was population-specific. This result suggests that the directions of selection can differ between populations connected by a low level of gene flow, which may contribute to the maintenance of high DRB allele diversity. In accordance with the optimality hypothesis, individuals with an intermediate number of alleles carried the lowest number of nematode species and had the lowest prevalence of A. tetraptera. However, the intensity of infection with A. tetraptera was linearly and negatively associated with the number of alleles.

The ecology of Bartonella spp. infections in two rodent communities in the Mazury Lake District region of Poland.

Prevalence and abundance of Bartonella spp. infections were studied over a 3-year period in woodland and grassland rodents in North-Eastern Poland. Prevalence of bacterial infections was similar in the two rodent communities, with one leading host species in each habitat (46.3% in Apodemus flavicollis versus 29.1% in Myodes glareolus in forest, or 36.9% in Microtus arvalis versus 13.7% in Mi. oeconomus in grassland). Prevalence/abundance of infections varied markedly across the 3 years with 2006 being the year of highest prevalence and abundance. Infections were more common during autumn months in My. glareolus and A. flavicollis, and in juvenile and young adult (age classes 1 and 2) My. glareolus and Mi. oeconomus than in adults (age class 3). Higher prevalence and abundance of Bartonella infections were found in male A. flavicollis in comparison to females. These data are discussed in relation to the parasite genotypes identified in this region and with respect to the role of various ecological factors influencing Bartonella spp. infections in naturally infected host populations.

Utilisation of rodent species by larvae and nymphs of hard ticks (Ixodidae) in two habitats in NE Poland.

The impact of host identity and habitat type on the density of hard ticks (Ixodes ricinus and Dermacentor reticulatus) infections on rodents in forest and abandoned field habitats in NE Poland was investigated. Ixodes ricinus was most abundant in the forest system, but D. reticulatus, although rarer, was most abundant in the field system. Environmental humidity and the much lower density of rodents probably limited the abundance of I. ricinus larvae in the field system, although this tick was still common on Microtus oeconomus from around small ponds. Nymphs of I. ricinus were comparatively rare in the forest, probably because of infection of non-rodent hosts. Dermacentor reticulatus nymphs on the other hand were very much more common in the ears of Microtus species than would have been predicted based on larval densities. The impact of habitat change (in this case successional change following field abandonment) on tick densities is emphasised, and the role of Apodemus as an epidemiological bridge, linking woodland and field habitats, is highlighted.

[Effect of coinfections in Ixodidae ticks on transmission of blood microparasites]. / Wplyw koinfekcji u kleszczy (Ixodidae) na transmisje mikro- pasozytów krwi.

The purpose of this review was to describe and discuss the current spectrum of coinfections in Ixodidae ticks and their effects on the transmission of blood microparasites. Coinfections with Borrelia burgdorferi s. l. and Anaplasma phagocytophilum and/or Babesia sp. in ticks from Poland appear to be common, however, the potential influence on transmission dynamics, the mechanism of genetic variation and the ecology of interactions between pathogens remain poorly understood compared with infections by single pathogen.

[Cryptosporidium spp. and Giardia spp.--environmental studies in Poland]. / Dwadziescia lat badan nad Cryptosporidium spp. i Giardia spp. w Polsce.

Cryptosporidium spp. and Giardia spp. are intestinal protozoan parasites of humans and many other species of mammals. The aim of this article was to summarize the last twenty years of research on the environmental distribution of these parasites, with a particular emphasis on the natural reservoir of invasion and human infections in Poland. The prevalence of Cryptosporidium and Giardia has been studied in different groups of humans, in wildlife, pets and farm animals and in environmental samples. Current knowledge on the distribution of zoonotic and non-zoonotic species/genotypes in reservoir hosts and environmental samples has been summarized. The usefulness of different methods for the detection and identification of the parasites in different types of samples has been presented. Due to the wide distribution and high prevalence of both species in a range of hosts and possible vectors involved in mechanical transmission, the overall risk of outbreaks of cryptosporidiosis and giardiosis in Poland has been assessed as relatively high.

Seasonality of the ectoparasite community of woodland rodents in a Mazurian Forest, Poland.

Myodes glareolus and Apodemus flavicollis support a large and diverse community of arthropod ectoparasites. This study of rodents in a Mazurian woodland sampled at monthly intervals between 2007 and 2009 revealed an ectoparasite community composed of 2 species of tick, 1 louse, 9 flea species, 7 species of gamasid mites, 4 fur mites and one trombiculid mite. A strong seasonality was noted in the dynamics of the ectoparasite community, with the fur mite Listrophorus sp. and the hypopus larva of a glycyphagid mite especially common in winter. Several of the ectoparasites have the potential to be pathogenic; the impact of these organisms on the population dynamics of their hosts remains to be investigated.

[Semi-aquatic animals as a source of water contamination with Cryptosporidium and Giardia]. / Zwierzeta ziemnowodne jako zródlo zanieczyszczenia wody Cryptosporidium i Giardia.

Cryptosporidium and Giardia spp. are parasitic protozoa localized in the alimentary tract of many animal species and humans. Each of these parasite species produces very resistant invasive forms (cysts and oocysts) excreted to the environment with feces of infected hosts. Water contaminated with cysts/oocysts constitutes one of the main transmission routes and is responsible for the majority of infections in humans. Cryptosporidium and Giardia spp. were found in many different species of animals, including livestock, pets and free living animals. The aim of our study was to determine the prevalence of these protozoa in selected species of semi-aquatic mammals and to estimate their role in water contamination. In years 1996-98 the prevalence of Cryptosporidium and Giardia infections was high in muskrats (Ondatra zibethicus) (58 and 87%, respectively). The origin of animals (farmed or free living) affected the prevalence of both parasites in European beavers (Castor fiber). The prevalence of infection increased in second period of study and was 4 and 19% for Cryptosporidium and 0 and 8% for Giardia spp. in the two studied periods, respectively. Both parasite species were also identified in water vole (Arvicola terrestris) and rat (Rattus norvegicus).

Cryptosporidium infection in patients with primary immunodeficiencies.

BACKGROUND: Cryptosporidium species infection is usually self-limited in immunocompetent populations, but can be severe and life-threatening among immunocompromised individuals, particularly in patients with AIDS and in these patients with primary immunodeficiencies (PIDs). PATIENTS AND METHODS: A group of 5 patients with genetically confirmed hyper-IgM syndrome type 1 (XHIM) and one patient with primary CD4 lymphopenia were enrolled in the study. At least 2 stool samples and a bile sample in one patient were examined for Cryptosporidium oocysts by a modified Ziehl-Neelsen technique, by immunofluorescence assay using a commercial kit, as well as by molecular analysis followed by genotyping. Immunological status at the time of PID diagnosis and the complex picture of disease are presented. RESULTS: Chronic cryptosporidiosis was confirmed in 3 patients with XHIM and in one patient with primary CD4 lymphopenia. Molecular diagnosis showed the presence of C parvum, C hominis, and C meleagridis in analyzed specimens. CONCLUSIONS: Cryptosporidium infection with serious clinical symptoms observed in patients with hyper-IgM syndrome calls for regular, repeated screening in this group of patients.

Long term monitoring of Babesia microti infection in BALB/c mice using nested PCR.

In this study we report the usefulness of nested PCR for screening of the persistent B. microti infections in rodent hosts. Female BALB/c mice were inoculated with 100 microl of donor blood infected with B. microti. Infections were detected using microscopic examination of Giemsa-stained blood smears. To determine whether B. microti DNA was present in blood and/or spleen tissue, nested PCR was performed targeting a specific fragment of the gene encoding the 18S rRNA. Blood was sampled every 10 days post-infection (dpi) until day 30, after which mice were sampled every 30 days until the end of experiment at 360 dpi. The most extensive parasitaemia (39% of infected erythrocytes) was observed at 10 dpi. Between 20-60 dpi, less then 1% of infected erythrocytes were detected in blood smears, and from 90 dpi onwards, infected erythrocytes were no longer observed. B. microti DNA was successfully amplified from the blood of mice from 10 dpi until 180 dpi, as well as from spleens of infected mice at 10 and 20 dpi. The presented results show that nested PCR is the method of choice for monitoring infections of B. microti in the blood of rodent hosts, and could therefore be a tool for environmental monitoring of naturally infected rodents which are the predominant source of infection for tick vectors.

Ticks and the city - are there any differences between city parks and natural forests in terms of tick abundance and prevalence of spirochaetes?

BACKGROUND: Ixodes ricinus ticks are commonly encountered in either natural or urban areas, contributing to Lyme disease agents Borreliella [(Borrelia burgdorferi (sensu lato)] spp. and Borrelia miyamotoi enzootic cycles in cities. It is an actual problem whether urbanization affects pathogen circulation and therefore risk of infection. The aim of the study was to evaluate main tick-borne disease risk factors in natural, endemic areas of north-east (NE) Poland (Bialowieza) and urban areas of central Poland (Warsaw), measuring tick abundance/density, prevalence of infection with spirochaetes and diversity of these pathogens in spring-early summer and late summer-autumn periods between 2012 and 2015. METHODS: Questing I. ricinus ticks were collected from three urban sites in Warsaw, central Poland and three natural sites in Bialowieza, NE Poland. A total of 2993 ticks were analyzed for the presence of Borreliella spp. and/or Borrelia miyamotoi DNA by PCR. Tick abundance was analyzed by General Linear Models (GLM). Prevalence and distribution of spirochaetes was analyzed by Maximum Likelihood techniques based on log-linear analysis of contingency tables (HILOGLINEAR). Species typing and molecular phylogenetic analysis based on the sequenced flaB marker were carried out. RESULTS: Overall 4617 I. ricinus ticks were collected (2258 nymphs and 2359 adults). We report well established population of ticks in urban areas (10.1 ± 0.9 ticks/100 m2), as in endemic natural areas with higher mean tick abundance (16.5 ± 1.5 ticks/100 m2). Tick densities were the highest in spring-early summer in both types of areas. We observed no effect of the type of area on Borreliella spp. and B. miyamotoi presence in ticks, resulting in similar prevalence of spirochaetes in urban and natural areas [10.9% (95% CI: 9.7-12.2%) vs 12.4% (95% CI: 10.1-15.1%), respectively]. Prevalence of spirochaetes was significantly higher in the summer-autumn period than in the spring-early summer [15.0% (95% CI: 12.8-17.5%) vs 10.4% (95% CI: 9.2-11.6%), respectively]. We have detected six species of bacteria present in both types of areas, with different frequencies: dominance of B. afzelii (69.3%) in urban and B. garinii (48.1%) in natural areas. Although we observed higher tick densities in forests than in maintained parks, the prevalence of spirochaetes was significantly higher in the latter [9.8% (95% CI: 8.6-11.0%) vs 17.5% (95% CI: 14.4-20.5%)]. CONCLUSIONS: Surprisingly, a similar risk of infection with Borreliella spp. and/or B. miyamotoi was discovered in highly- and low-transformed areas. We suggest that the awareness of presence of these disease agents in cities should be raised.

Abundance of wild rodents, ticks and environmental risk of Lyme borreliosis: a longitudinal study in an area of Mazury Lakes district of Poland.

The results of a longitudinal epidemiological survey in two contrasting habitats in an area of the Mazury Lakes district of Poland indicate that both host and vector (Ixodes ricinus) densities, may be the most important risk factors for the tick-transmitted spirochetes of Borrelia burgdirferi s.l. However, the results also highlight that even related host species, such as the wild rodents Apodemus flavicollis and Clethrionomys glareolus that share the same habitat, can show quite different dynamics of tick infestation. We provide evidence that the woodland populations of A. flavicollis and C. glareolus are more frequently infested with larvae than nymphs, and more frequently with both stages than M. arvalis in the neighbouring open fallow lands. The prevalence of infestation with larvae varied from 92 % for A. flavicollis, and 76 % for C. glareolus to 37 % for M. arvalis. Other factors, such as population age structure and sex, were also shown to impact on tick densities on hosts at particular times of the year and hence on the zoonotic risk. Moreover, particular species of rodents from different habitats, A. flavicollis (woodlands) and Microtus arvalis (fallow lands) carry infected immature I. ricinus ticks more frequently than C. glareolus voles (woodlands). Thus, the relative contribution of each species to the cumulative reservoir competence differs among species living in the woodland habitats and in relation to voles living in the fallow lands. It follows, therefore, that any factor which reduces the relative density of A. flavicollis in comparison to other hosts in the wild rodent community, will reduce also the risk of human exposure to Lyme borreliosis spirochetes.

Borrelia miyamotoi: A human tick-borne relapsing fever spirochete in Europe and its potential impact on public health.

Borrelia miyamotoi is a tick-borne bacterium which has only recently been identified in Europe as a human pathogen causing relapsing fever and little is known about its local impact on human health. There are three types of B. miyamotoi: Asian (Siberian), European, and American. B. miyamotoi is transmitted by the same Ixodes ricinus-persulcatus species complex, which also transmits B. burgdorferi s.l., the Lyme borreliosis group. Both Borrelia groups are mostly maintained in natural rodent populations. The aim of this review is to summarize the available literature on B. miyamotoi, with the focus of attention falling on Europe, as well as to describe its presence in ticks, reservoir hosts, and humans and discuss its potential impact on public health.