<Emphasis Type="Italic">Dermacentor reticulatus</Emphasis> ticks (Acari: Ixodidae) distribution in north-eastern Poland: an endemic area of tick-borne diseases

Dermacentor reticulatus is the second most important tick species in Poland. Although the north-eastern region of Poland is considered typical for D. reticulatus and is treated as a contiguous area of Eastern populations of the meadow ticks, the occurrence of this tick species in this region to date has been recorded as separate foci. The present report supplements data on the geographical distribution of D. reticulatus in urban and natural biotopes of north-eastern Poland (Warmia and Mazury province). In 2015–2017 (during the springtime activity of ticks) adult questing D. reticulatus were found in 13 of 25 monitored localities. Six sites are located in urbanized areas, within the administrative borders of the city of Olsztyn and seven sites are in natural biotopes in the central part of Warmia and Mazury. A total of 398 adult D. reticulatus ticks, including 257 females and 141 males, were collected. A comparison of data grouped according to urban and natural type of area revealed no statistical differences between them. Taking into account the habitat type, the mean tick density was the highest in open landscapes. The identification of new foci D. reticulatus in the endemic areas of Lyme borreliosis, tick-borne encephalitis and canine babesiosis is crucial for determining the risk of diseases transmitted by ticks and taking proper preventive measures.     

Discovery of <Emphasis Type="Italic">Dermacentor reticulatus</Emphasis> (Acari: Amblyommidae) populations in the Lubuskie Province (Western Poland)

The distribution of the meadow tick, Dermacentor reticulatus (Fabricius) is divided into two separate areas—Western Europe and Eastern (Russian). The break in distribution of this species falls in Poland, as well as other sites. The populations of D. reticulatus in Poland, which have been confirmed by collecting ticks from vegetation, are located in the north-eastern and western parts of the country, although there are also reports of D. reticulatus occurring on hosts in several parts of Poland which have not been confirmed by collecting ticks from vegetation. Until this research was carried out, the view was that this species does not occur in the western part of Poland in the area between the Vistula and Oder rivers. In the site surveys conducted in the Lubuskie Province (western part of Poland around the city of Zielona Góra, about 55 km from the Polish-German border), during the springtime activity peak for adult stages looking for hosts, seven natural implantations of D. reticulatus were discovered where 208 tick specimens were collected by flagging. These were exclusively adult stages, including 127 females and 81 males. Because of the distance (around 350 km) from the nearest focus in Mazuria in North-eastern Poland, the populations discovered have been named the D. reticulatus “Lubuskie Focus”. The research proves for the first time that D. reticulatus occurs in Western Poland. The research indicates that D. reticulatus is the second most important tick in Poland, after Ixodes ricinus (L.), which is epidemiologically very important, especially as a parasite on dogs and cattle. The proliferation of the meadow tick in Central Europe is inseparably linked with the expansion of canine babesiosis.     

Tick-borne pathogens in Dermacentor reticulatus collected from dogs in eastern Poland

In recent years, the distribution of Dermacentor reticulatus ticks has expanded into new territories in many European countries, including Poland, with increased population densities in areas of their regular occurrence. The spread of D. reticulatus enhances the risk of exposure of domestic animals and their owners to tick-borne diseases. The objective of this study was to assess the prevalence of infection of D. reticulatus ticks feeding on dogs with the pathogens Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum. The study material comprised 152 D. reticulatus ticks collected from dogs in the northeastern part of Lublin Province (eastern Poland). A ready-made AmpliSens® TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis/E.muris-FRT PCR kit was used for qualitative detection and differentiation of tick-borne infections. The assessment of the degree of infection of the analyzed ticks with the two pathogens revealed that 9.2% (14/152) of the examined ticks were infected with one of the pathogens. No co-infections with the pathogens were detected in any of the ticks. The highest specific percentage of infections (8.6%, 13/152) was associated with A. phagocytophilum. The presence of B. burgdorferi s.l. was detected in only one of the examined ticks (0.7%). The spread of D. reticulatus to new territories and the increase in population density in areas of their regular occurrence implies the need for further studies of the prevalence of pathogens with medical and veterinary importance in order to assess the risk of tick-borne diseases.

     

Detection of Borrelia burgdorferi s.l., Anaplasma phagocytophilum and Babesia spp. in Dermacentor reticulatus ticks found within the city of Białystok,Poland—first data

Pathogens carried by ticks pose a threat to both human and animal health across the world. Typically associated with rural landscapes, ticks appear to adapt well to life in urban recreational areas. Although Dermacentor reticulatus is commonly found across Europe, data on the prevalence of pathogens in this tick species, in an urban environment, are very limited. PCR was used to examine 368 D. reticulatus individuals collected in the Zwierzyniecki Forest Nature Reserve in Bia?ystok, Poland. In total, 10.3% of ticks were infected, with Babesia spp. (9.2%), Anaplasma phagocytophilum (0.8%) and Borrelia burgdorferi sensu lato (0.3%). Rickettsia spp., Bartonella spp., and Coxiella burnetii were not detected. Sequence analysis for Babesia-positive samples identified 79.4% of them as Babesia canis, 8.8% as Babesia microti, 5.9% as Babesia spp., 2.9% as Babesia venatorum, and 2.9% as Babesia vogeli. Results obtained in this study indicate that D. reticulatus ticks found within the urban premises of the study area are infected with at least three pathogens and therefore are an important factor in public health risk for tick-borne diseases.

     

Prevalence of tick-borne encephalitis virus (TBEV) RNA in Dermacentor reticulatus ticks from natural and urban environment,Poland

Tick-borne encephalitis virus (TBEV) (Flaviviridae, Flavivirus) is an arthropod-borne virus, an etiologic agent of tick-borne encephalitis (TBE), a human infection involving the central nervous system. The disease is endemic in a large region in Eurasia, where it is transmitted mainly by Ixodes ricinus and Ixodes persulcatus ticks. It is known that also Dermacentor reticulatus is involved in circulation of TBEV. However, the current knowledge of D. reticulatus importance in TBE epidemiology is still insufficient. A total of 471 adult D. reticulatus ticks were collected by flagging vegetation in the Bia?owie?a Primeval Forest, Biebrza National Park, Masurian Landscape Park (North-Eastern Poland) and in the city of Warsaw in the years 2007–2010. All collected ticks were examined individually for the presence of RNA of TBEV using nested RT-PCR assay. Positive results were noted in all investigated localities with the infection rate ranging from 0.99 to 12.5 % with a total mean of 2.12 %. The difference in the percentage of infective males and females was not statistically significant.     

Phenology of ixodid ticks of the genus Dermacentor in the Central Ciscaucasia

In Stavropol Territory, ixodid ticks of the genus Dermacentor are represented by three species: D. marginatus, D. reticulatus, and D. niveus. D. marginatus is the most widespread species, constituting 14.8% of all the ticks in the region; D. reticulatus and D. niveus are found less frequently, constituting 2.8% and 0.01%, respectively. D. marginatus and D. reticulatus are spread mostly in the forest-steppe and steppe zones of Stavropol Territory. D. niveus is represented by solitary specimens in arid and semidesert zones. The main hosts of adult D. marginatus in Stavropol Territory are cattle (88.4%) and sheep and goats (5.3%). The hosts of D. reticulatus are also cattle (72.3%) and sheep and goats (4%). D. marginatus activates at an average monthly temperature of +2.07°C and average monthly rainfall of 47.2 mm. The peak of parasitizing on hosts was observed at an average monthly temperature of +7.8°C and average monthly rainfall of 56.2 mm. The autumnal activation was recorded at an average monthly temperature of +15.3°C and average monthly rainfall of 58.2 mm.     

Evidence for the reproductive isolation ofDermacentor marginatus andDermacentor reticulatus (Acari: Ixodidae) ticks based on cross-breeding, morphology and molecular studies

The species status ofDermacentor marginatus andDermacentor reticulatus was evaluated by scanning electron microscope (SEM) examination of adult ticks, cross-breeding experiments and molecular biological analysis of eggs derived from transspecific pairings. The SEM investigations including the morphometric quantification of phenotypic features, resulted in an unequivocal differentiation of adultD. marginatus andD. reticulatus ticks. The cross-breeding experiments demonstrated that irrespective of whether female ticks of both species were applied with con- or transspecific male ticks or without males to sheep, they engorged and laid eggs. The larvae, however, developed only in eggs which originated from conspecific matings. A nested polymerase chain reaction (PCR) of the second internal transcribed spacer (ITS2) using the DNA of eggs from transspecific pairings and sequencing of the PCR products revealed two different genotypes. The genotypes of eggs originating fromD. marginatus andD. reticulatus females of these pairings differed. However, the eggs deposited byD. marginatus always possessed the same two genotypes as did the eggs produced byD. reticulatus. These results argue for a strict reproductive isolation ofD. marginatus andD. reticulatus and, therefore, for a separate species status.     

The role of rock mining for maintaining Dauco carotae-Crepidetum rhoeadifoliae Hejný et Grüll in Hejný et al. 1979 — a new to Poland plant association

This work presents the Dauco carotae-Crepidetum rhoeadifoliae plant association, which is new to Poland. The association has been observed in industrial reclamation areas in the vicinity of carbonate mineral excavation sites in the central part of the Opole region. In the vast majority of cases, plots of this association developed in reclaimed areas. The majority of diagnostic species for the association was found within surveyed plots, including Verbascum thapsus, V. densiflorum and Bryum argenteum. Taxa characteristic of the alliance were also constantly present, i.e. Daucus carota, Melilotus alba, M. officinalis, Echium vulgare and Erysimum hieracifolium. This association belongs to the rarest syntaxa in Poland included in the Dauco-Melilotion alliance of ruderal communities with a predominance of hemicryptophytes, therophytes and perennials. The main diagnostic species — Crepis rhoeadifolia, belongs to very rare elements of Polish flora. It has been observed only in the southern part of the country in approx. 20 sites. Crepis rhoeadifolia had not been observed in Silesia for approx. 40 years, which is why it was considered to be an extinct taxon in this region. Rediscovering of the species allowed for diagnosing the Dauco-Crepidetum rhoeadifoliae association. This association is an example of a pioneer phytocenosis of, most likely, anthropogenic origin in Silesia.     

Prevalence of Tick-Borne Pathogens in Ixodes ricinus and Dermacentor reticulatus Ticks from Different Geographical Locations in Belarus

Worldwide, ticks are important vectors of human and animal pathogens. Besides Lyme Borreliosis, a variety of other bacterial and protozoal tick-borne infections are of medical interest in Europe. In this study, 553 questing and feeding Ixodes ricinus (n = 327) and Dermacentor reticulatus ticks (n = 226) were analysed by PCR for Borrelia, Rickettsia, Anaplasma, Coxiella, Francisella and Babesia species. Overall, the pathogen prevalence in ticks was 30.6% for I. ricinus and 45.6% for D. reticulatus. The majority of infections were caused by members of the spotted-fever group rickettsiae (24.4%), 9.4% of ticks were positive for Borrelia burgdorferi sensu lato, with Borrelia afzelii being the most frequently detected species (40.4%). Pathogens with low prevalence rates in ticks were Anaplasma phagocytophilum (2.2%), Coxiella burnetii (0.9%), Francisella tularensis subspecies (0.7%), Bartonella henselae (0.7%), Babesia microti (0.5%) and Babesia venatorum (0.4%). On a regional level, hotspots of pathogens were identified for A. phagocytophilum (12.5–17.2%), F. tularensis ssp. (5.5%) and C. burnetii (9.1%), suggesting established zoonotic cycles of these pathogens at least at these sites. Our survey revealed a high burden of tick-borne pathogens in questing and feeding I. ricinus and D. reticulatus ticks collected in different regions in Belarus, indicating a potential risk for humans and animals. Identified hotspots of infected ticks should be included in future surveillance studies, especially when F. tularensis ssp. and C. burnetii are involved.     

The oldest nautiloid recorded from the Triassic Germanic Basin and the first ammonoid from the Aegean (Middle Triassic) of Poland

During the Triassic in Europe, the Germanic Basin extended from England in the west to the eastern border of Poland in the east. Although cephalopods are common in some Middle Triassic (Muschelkalk) horizons, there still persists a gap in the palaeontological record of the eastern part of the Germanic Basin, notably in the lowermost parts of the Lower Muschelkalk, spanning the latest Olenekian-Aegean interval. The present contribution attempts to fill this gap by presenting the first ammonoid (ceratitid Beneckeia sp.) and nautiloids (Germanonautilus cf. dolomiticus and G. cf. salinarius) from the Lower Gogolin Beds (Upper Silesia, Poland). These Germanonautilus are the oldest nautiloids found in the entire Germanic Basin.     

Studies on survival and water balance of unfed adult Dermacentor marginatus and D. reticulatus ticks (Acari: Ixodidae)

The water content, the survival time at various relative humidities(r.h.) and the critical equilibrium activity of unfed adultDermacentor marginatus and D.reticulatus ticks were investigated at a constant temperature of20 °C. It was also examined whether these ticks use liquidwaterto compensate water loss. Both Dermacentor spp. showed nosignificant differences in water content in relation to body mass. The meanwater content of D. marginatus and D.reticulatus was 54.6% and 54.7%, respectively, in females and 56.3%and 57.0%, respectively, in males. The survival time of unfed adults prolongedwith decreasing saturation deficits. On average, males survived longer thanfemales and D. marginatus ticks survived mostly longerthanD. reticulatus ticks. The 50% mortality period rangedbetween 40 d at 33% r.h. and 420 d at 95% r.h. in D.marginatus, and between 43 d at 33 r.h. and 366 d at 95% r.h. inD. reticulatus. The critical equilibrium activity of unfedadults was estimated to be 0.84 for both species and was independent of sex.When dehydrated adult D. marginatus and D.reticulatus ticks were offered liquid water, only a few slightlygained weight while most further lost weight. Liquid water was not attractivefor dehydrated or non-dehydrated ticks and drinking was not observed. Aftersubmerging in water for 2 d, most of the dehydrated ticks had gained weight.     

Molecular detection of Babesia canis in Dermacentor reticulatus ticks collected in Slovakia

Dermacentor reticulatus ticks are recognized as the most important vectors of Babesia canis, the aetiological agent of canine babesiosis occurring throughout Europe. Vector competence of D. reticulatus for B. canis is well described and experimentally determined; however, by using molecular analysis it was proven so by one recent study in Russia. Herein, the additional molecular evidence of B. canis infection in D. reticulatus ticks collected in Slovakia is provided. Using PCR followed by sequencing of distinctive amplicons we determined the presence of Babesia canis canis in one of 100 tested adult ticks. Two zoonotic pathogens, Francisella tularensis and Coxiella burnetii, were previously isolated from D. reticulatus ticks in Slovakia. In our samples, we detected only the presence of F. tularensis.     

Studies on the critical water mass and the rehydration potential of unfed adult Dermacentor marginatus and D. reticulatus ticks (Acari: Ixodidae)

In unfed adult Dermacentor marginatus and D. reticulatusticks survival and capability to restore water balance after loss of high percentages of exchangeable body water were investigated. Furthermore, it was examined how frequently dehydrated ticks of these species were able to rehydrate by uptake of atmospheric water vapour. The critical water mass, defined as the water mass remaining in a tick at the nonambulatory state, differed between light and heavy weight groups and averaged 62.4 and 55.8%, respectively, of the total body water of fully hydrated ticks in females, and 54.4 and 51.1% respectively, in males of D. marginatus. In D. reticulatus, the corresponding figures were 55.9 and 54.7% in females and 52.1 and 52.7% in males. All ticks survived dehydration to 50, 75 or 100% of the critical water mass, and 96.7% of the D. marginatus ticks and 95.8% of the D. reticulatus ticks compensated water losses during subsequent incubation at 95% relative humidity (r.h.) and 20°C. Unfed females and males of both Dermacentor spp. were capable to balance water loss very frequently over a period of several months. When ticks were repeatedly dehydrated at 0% r.h. for 7 days and rehydrated at 95% r.h. and 20°C, females and males of D. marginatus reached the 50% mortality after 22 and 29 cycles of de- and rehydration, respectively, during 211 and 285 days, respectively. In D. reticulatus, 50% of females and males survived 23 and 17 cycles, respectively, during 248 and 186 days, respectively. Rehydration weights were as high or even higher as those of ticks kept at permanent 95% r.h.     

Molecular identification and biodiversity of potential allergenic molds (Aspergillus and Penicilium) in the poultry house: first report

The production of poultry has proved to be a significant source of fungi. Penicillium and Aspergillus are among the most important species, because they can cause irritations, infections, allergies, spoilage of food and beverages and are able to produce dangerous mycotoxins. Detection and identification of the species in these genera is very significant, because it provides relevant information about the properties of the responsible strains. Therefore, their taxonomic was determined using a phenotypic and molecular (ITS and partial β-tubulin sequences) methods. Investigations were conducted in the poultry house located near Wroc?aw in Lower Silesia (Poland). Using morphological analyses, 215 fungal strains were identified. Among them, 56 belonged to the Penicilium and Aspergillus genera. The results obtained from sequence analysis corresponded well with the morphological identification. Classification at species level was possible in most cases using the sequence data. Most isolates belong to Penicilium genus, with the follow dominating species (P. chrysogenum, P. polonicum and P. olsonii).     

Cultivable internal bacterial flora of ticks isolated in Hungary

From six sampling sites in north-western Hungary 126 questing ticks of three species (Ixodes ricinus, Dermacentor reticulatus, Haemaphysalis concinna) were sampled. After inactivating the external bacteria on the surface of ticks, the internal bacterial flora was cultured (two kinds of agar media at three temperatures with aerobic and anaerobic conditions were applied), and 116 strains were isolated. Our results showed, that after a blood meal the bacterial contamination of ticks was much higher in Ixodes and Dermacentor, than in Haemaphysalis, indicating different host range or blood meal habits. Most (89.7 %) of the bacteria were Gram-positive, the most frequent genera were the Staphylococcus (18.1 %) and Bacillus (7.8 %). The percentage of bacteria part of the skin and mucosal flora was 21.6 %. Among the environmental bacteria 14 were found with reported medical importance. The results show, that members of some genera are able to replicate inside the ticks (Mycobacterium, Bacillus) which can increase their potential risk. Isolated bacteria/tick ratio continuously grew from larvae to adults, indicating that larvae probably are hatched sterile, but later bacterial uptake from the environment and from the hosts increases bacterial contamination. Ten anaerobic bacteria were cultured, mostly Propionibacterium acnes, a facultative skin pathogen. No significant differences were found between the isolated bacteria of the six sampling sites. Our work showed, that internal bacterial community of ticks is diverse, novel strains were isolated several with medical importance, some bacteria multiplicate inside ticks, and that ticks continuously take up bacteria from the environment. Our study first described anaerobic bacteria from ticks.     

Isolation ofNosema slovaca (Microsporidiae) fromDermacentor reticulatus ticks (Acari: Ixodidae) collected in Hungary

Nosema slovaca (Microsporidiae) was isolated from a maleDermacentor reticulatus tick collected in the Mélykút locality, Bács-Kiskun province, Hungary. This parasitic protozoan caused an acute infection in partly engorgedD. reticulatus females when inoculated intracoelomically resulting in death of the ticks within 5–15 days post-infection. This strain ofNosema can be maintained in the laboratory by passaging between partly engorgedD. reticulatus females and is currently being studied for its potential as a biological control agent of ixodid ticks.     

Abundance and seasonal activity of questing Ixodes ricinus ticks in their natural habitats in southern Germany in 2011

Questing ticks were sampled monthly over a period of 11 months from February, 2011 to December, 2011 at 13 sites in southern Germany using the flagging method. The ticks were identified to species, gender, and stadium. Although both I. ricinus and D. reticulatus were sampled, this study concentrated on I. ricinus, since it was the most abundant tick to be found. Additional weather data (air and soil temperature, relative air humidity, precipitation, sunshine duration) were recorded on each sampling site and the local vegetation described. A total of 14, 394 ticks was collected (7,862 larvae, 5,568 nymphs, 964 adults) and their activity was recorded in order to determine the seasonal activity pattern over different periods of the year. In contrast to the widely accepted pattern of a bimodal seasonal activity in moderate areas with a dominant peak in spring and a minor peak in autumn, a unimodal activity pattern was found for all development stages on six of the 12 sampling sites. Tick abundance was compared to weather variables. Tick host‐seeking activity was found to be significantly dependent on the temperature at ground level, precipitation, and sunshine duration as well as relative air humidity. Adult ticks showed a positive correlation with the duration of sunshine, whereas nymphs were mostly unaffected by this phenomenon.     

Francisella tularensis prevalence and load in Dermacentor reticulatus ticks in an endemic area in Central Europe

A total of 7778 host‐seeking adult Dermacentor reticulatus (Ixodida: Ixodidae) ticks were examined for the prevalence of Francisella tularensis holarctica (Thiotrichales: Francisellaceae) in a natural focus of tularaemia in the floodplain forest–meadow ecosystem along the lower reaches of the Dyje (Thaya) river in South Moravia (Czech Republic) between 1995 and 2013. Ticks were pooled (10 specimens per pool) and their homogenates inoculated subcutaneously in 4‐week‐old specific pathogen‐free mice. Dead mice were sectioned, their spleens cultivated on thioglycollate–glucose–blood agar and impression smears from the spleen, liver and heart blood were Giemsa‐stained. Sixty‐four pools were positive for F. tularensis: the overall minimum infection rate (MIR) was 0.82%. Overall MIRs for the 4714 female and 3064 male D. reticulatus examined were 0.89 and 0.72%, respectively; MIRs fluctuated across years between 0.0 and 2.43%. The estimated bacterial load in infected ticks varied from 0.84 to 5.34 log₁₀ infectious F. tularensis cells per tick (i.e. from about seven to 220 000 cells). Ticks with low loads were more prevalent; more than 1000 infectious cells were detected in 24 ticks (0.3% of all ticks and 37.5% of infected ticks). Monitoring of D. reticulatus for the presence and cell numbers of F. tularensis may be a valuable tool in the surveillance of tularaemia.     

Influence of selected meteorological variables on the questing activity of Ixodes ricinus ticks in Lower Silesia,SW Poland

The relationship between climate data and tick questing activity is crucial for estimation of the spatial and temporal distribution of the risk of ticks and tick‐borne diseases. This study establishes correlations between selected meteorological variables provided by the Weather Research and Forecasting model (WRF) and the questing activity of Ixodes ricinus nymphs and adults on a regional scale across Lower Silesia, Poland. Application of Generalized Linear Mixed Models (GLMM), built separately for adults and nymphs, showed that solar radiation, air temperature, and saturation deficit appeared to be the meteorological variables of prime importance, whereas the wind speed was less important. However, the effect of meteorological parameters was different for adults and nymphs. The adults are also more influenced by forest cover and the percentage of forest type if compared to nymphs. The WRF model providing meteorological variables separately for each location and day of tick sampling can be useful in studies of questing activity of ticks on a regional scale.     

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.