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Article

Altitudinal Distribution, Seasonal Dynamics and Borrelia burgdorferi Sensu Lato Infections in Hard Ticks (Acari: Ixodidae) in Different Forest Communities in Inland Croatia

by
Marko Vucelja
1,*,
Stjepan Krčmar
2,*,
Josipa Habuš
3,
Vesna Mojčec Perko
3,
Marko Boljfetić
4,
Linda Bjedov
1 and
Josip Margaletić
1
1
Faculty of Forestry and Wood Technology, University of Zagreb, 10000 Zagreb, Croatia
2
Department of Biology, Josip Juraj Strossmayer University of Osijek, 31000 Osijek, Croatia
3
Department of Microbiology and Infectious Diseases, with Clinic, Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia
4
Oikon Ltd.—Institute of Applied Ecology, 10020 Zagreb, Croatia
*
Authors to whom correspondence should be addressed.
Sustainability 2023, 15(6), 4862; https://doi.org/10.3390/su15064862
Submission received: 13 January 2023 / Revised: 7 March 2023 / Accepted: 7 March 2023 / Published: 9 March 2023
(This article belongs to the Special Issue Forest Operations and Sustainability)
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Abstract

:
Altitudinal distributions, population structures and seasonal dynamics of tick fauna at three localities in Continental Croatia (Medvednica and Papuk) and an alpine biogeographic region (Gorski Kotar) were studied. Sampling of questing ticks was performed twice a year (spring and autumn, from 2019 to 2021) at different altitudes (200, 400, 600, 800 and 1000 m above sea level) using the flagging method. In total, 2942 ticks (53.9% larvae, 40.1% nymphs, 6.0% adults) were sampled and 2937 (99.83%) were determined as Ixodes ricinus, 4 (0.14%) as Haemaphysalis concinna and 1 (0.03%) as Ixodes frontalis. Ixodes ricinus was the only species found at all altitudes and sampling sites. The highest tick abundance was recorded at higher altitudes (800–1000 m asl.) on Medvednica and in Gorski Kotar within mixed forests of European beech and European silver fir, while on Papuk most of the ticks were sampled at lower altitudes (200 m asl.) in Sessile oak forest. From 27 pools containing 305 ticks, 1 (3.7%) was positive for Borrelia burgdorferi sensu lato infection. Borrelia burgdorferi s.l. was detected in 20% (1/5) of the pools containing nymphs and adults collected in Gorski Kotar (600 and 800 m asl.). Sequencing of the ospA gene and phylogenetic analysis revealed the presence of the Borrelia burgdorferi sensu stricto genotype.

1. Introduction

Hard ticks (Acari: Ixodidae) are obligatory hematophagous ectoparasites and one of the main arthropod-borne disease vectors worldwide. More than a century ago, they were the first arthropods proved as vectors of infectious disease [1,2]. Tick-borne diseases (e.g., Lyme borreliosis, tick-borne encephalitis, tularemia, Mediterranean spotted fever and human granulocytic anaplasmosis) are extremely diverse, while infectious agents transmitted by ticks include numerous bacteria, viruses and protozoa [3]. As ticks and tick-borne diseases (TBDs) infest a wide variety of wild vertebrate species, as well as livestock, companion animals and humans, they are also a cause of significant economic losses worldwide [4]. In recent decades, ticks have gained more public attention due to the expansion of their (altitudinal and latitudinal) ranges and the increasing incidence of tick-borne diseases [5,6,7]. Such increase is a result of different environmental and social factors, e.g., climate changes (prolonged growing seasons, warmer and shorter winters, etc.) [8,9,10], loss of predators [11], reforestation of agricultural lands, woodland expansion [12,13], urbanization, human mobility and population growth [14,15,16]. The higher risk of TBDs is typically associated with woodland habitats that provide ticks with the most favorable microclimatic conditions and high availability of hosts [17,18,19]. Even so, a rising number of medically important tick species populations are also being recorded in urban and suburban areas across Europe [10]. One tick species well-known for its high abundance and wide distribution in Europe and Croatia is Ixodes ricinus (Castor bean tick, Sheep tick, Deer tick, Forest tick) [20]. It is a major vector for a great number of pathogens (e.g., Borrelia burgdorferi sensu lato, Borrelia miyamotoi, tick-borne encephalitis virus, Rickettsia spp., Anaplasma phagocytophilum, Babesia divergens, Babesia microti and Francisella tularensis) [21,22]. Ixodes ricinus is the main vector of Borrelia burgdorferi s.l. in inland Croatia [23,24], especially in the northern and northwestern parts, where high prevalences (40–86%) of Borrelia burgdorferi have been recorded within tick populations [25,26,27]. Lyme borreliosis or Lyme disease, recognized in 1975 in the town of Old Lyme, Connecticut, in North America [28,29], is currently the most common tick-borne and vector-borne disease in Europe and in the USA [30,31]. Borrelia burgdorferi sensu lato (B. burgdorferi s.l.) is a bacterial complex transmitted by hard ticks that contains multiple genospecies. Of 21 genospecies identified, 8 are present in Europe: B. burgdorferi sensu stricto (s.s.)., B. afzelii, B. garinii, B. bissettiae, B. bavariensis, B. spielmanii, B. lusitaniae and B. valaisiana [32], and the first 6 cause disease in humans. The incidence of Lyme borreliosis has more than doubled over the last decades in Europe [33]. In Croatia, it has shifted from 255 to more than 400 reported human cases per year in the last 20 years [34]. It is recognized now that Borrelia burgdorferi exists at quite variable rates wherever the vector exists and that maximum prevalence is recorded in large regions of Central Europe, where the infection rates of Ixodes ricinus ticks range from 3% to 45% [10,35,36]. While there is an environmental niche that accounts for the prevalence of the causative agents, such as Borrelia burgdorferi s.l. [36], there is a necessity for a deeper understanding of tick fauna and their zoonotic potential in our rapidly changing environment. In their recent review, Kahl and Gray (2023) pointed out that over the last 20 years there were 2.4 times more publications on ticks and tick-borne diseases than in the previous 20 years [37]. The latter is not so surprising given that vector-borne diseases have been recognized as some of the most important public health problems of this century [10]. The most comprehensive ecological studies on the diversity of tick fauna in Croatia have focused mainly on the Mediterranean region (the Adriatic Coast, along with the islands known for their karst topography) and they were conducted during the second half of the twentieth century [20,38,39,40,41,42]. Croatian northwestern and western regions were only partly included [26,43,44]. Novel studies, carried out during the last two decades, have mainly been made at lower altitudes in central and eastern parts of inland Croatia [45,46,47]. A total of 23 hard tick species, belonging to five genera (Ixodes, Haemaphysalis, Rhipicephalus, Dermacentor and Hyalomma) have been recorded in Croatia so far [20,48]. There have been no studies on the altitudinal distributions of ticks in mountainous regions of Croatia for various reasons (i.e., terrains being unreachable by vehicles, small human populations, etc.). This research dealt with the question of the diversity of tick fauna, their spatio-temporal distribution and the zoonotic threat in the continental and alpine regions of Croatia. The aim of this study was to determine tick species, their altitudinal distributions, population structures, seasonal dynamics and the prevalence of Lyme disease causative agents (Borrelia burgdorferi s.l.) within tick fauna at three mountain sites in inland Croatia. We hypothesized that the most prevalent tick species would be from the genera Ixodes and Dermacentor, that they would appear in higher abundances during springtime sampling, that their numbers would decrease with higher altitudes and also that the highest prevalences of Borrelia burgdorferi s.l. would be found within tick populations on the Medvednica mountain in the northwestern part of the country, as it was previously known as a Lyme disease focus in Croatia.

2. Study Area

In this study, research on tick fauna was carried out from 2019 to 2021 at three locations within two biogeographic regions in inland Croatia [49]. Tick species diversity and their altitudinal distributions, population structures and seasonal dynamics were monitored at two mountain sites (Medvednica and Papuk) in the continental biogeographic region and one site in the alpine region (Gorski Kotar) (Figure 1).
According to the geographical distribution of climatic types by Köppen, all three mountain study sites belong to Cfb—a temperate climate without a dry season and with a warm summer [50,51]. The temperature of the coldest month of the year (January) is above −3 °C, while the summers are fresh, with the average monthly temperature of the hottest month being below 22 °C [52]. Nomenclature of forest communities and soil types correspond to data specified in “Forest phytocenology and forest communities in Croatia” by Vukelić and Rauš (1998) and “Croatian forest vegetation” by Vukelić (2012) [53,54] and “Management of forest soils in Croatia” by Martinović, respectively [55]. Overall, the study included 13 sampling sites and nine different forest communities. The list of tick sampling sites, their GPS positions, altitudes and forest communities are shown in Table 1. Photography of every sampling site is given in Appendix A (Figure A1, Figure A2, Figure A3, Figure A4, Figure A5, Figure A6, Figure A7, Figure A8, Figure A9, Figure A10, Figure A11, Figure A12 and Figure A13).
Tick sampling was performed from 2019 to 2021, twice a year (spring and autumn), at all three mountain sites at different altitudes. Mean air temperatures and air humidity data for the spring (March–June) and autumn (September–December) sampling seasons are shown in Table 2. The data from the meteorological stations nearest to the sampling locations were provided by the Croatian Meteorological and Hydrological Service.
The area of Gorski Kotar covers 1275 square kilometers, and it is located in the zone of Central European climate characterized by long and snowy winters, low average annual temperature, high air humidity and abundant precipitation with quite strong winds from the northeast and southwest. The flora and fauna of Gorski Kotar are known for their richness, while 63% of the total area of Gorski Kotar is covered by forests (“Green lungs of Croatia or Croatian Switzerland”), with a predominance of fir–beech forest communities. The highest peak is Bjelolasica (1534 m).
Medvednica (pronounced [mědυednitsa], lit. “Bear Mountain”) is a mountain area just north of Croatia’s capital, the city of Zagreb. It extends from the southwest to the northeast and is 42 km in length. In 1981, it was declared a nature park with a protected area of 228 km2. The plant cover of Medvednica is mostly represented by natural and preserved forests (63% of the area). Due to the uneven terrain, various geological substrates and soil types, as many as 12 forest communities appear there, with distinct zonations depending on altitude and exposure. The predominant species of trees are European beech, holm oak, hornbeam, sweet chestnut, maple and ash. Medvednica is a home to various types of mammals: roe deer, wild boar, wild cat, fox, marten and weasel. The highest point is Sljeme (1033 m asl).
Papuk is a mountain in Eastern Croatia, on the northern and northwestern border of the Požega basin. In 1999, the the Papuk Nature Park was founded, and it encompasses the largest part of the Papuk mountain (33,600 ha), with a general NW-SE extension of about 45 km. Forest habitats, with 11 different forest communities, cover around 95% of the park. The dominant tree species are European beech (47%), sessile oak, fir, hornbeam, sycamore, Austrian oak, pubescent oak, wild cherry, etc. The dense forests of Papuk are the habitats of deer, roe deer, wild boars, foxes and martens. Its highest peak, also named Papuk, is 953 m asl.

3. Materials and Methods

3.1. Tick Sampling and Identification

Ticks were sampled by the flag dragging method [56,57,58], using white flannel cloth (1 m × 1 m) on two transects (2 × 100 m in length) at every sampling site. The flag was pulled over the ground surface, over leaves and low vegetation, and it was inspected every 5–10 m, depending on the catch. Ticks were collected from the flag with tweezers and stored (alive) in plastic tubes (eppendorf, 1.5 mL) with a safety cap.
During the field work, a mobile application and the online database “krpelji.info” (https://www.krpelji.info/) (accessed on 5 December 2021) were used while sampling ticks. The application was developed (and is still in “test mode”) by the Faculty of Forestry and Wood Technology (FFWT), University of Zagreb. It allows the recording of GPS positions (waypoints and routes/transects), altitudes, date and time of sampling, vegetation type, sampling methods, number of collected specimens, tick species and their life stages. Additional uses of the application included photographing the sampling site, collected ticks, etc.
The sampled ticks were transported to FFWT, where a visual determination of the ticks and their life stages (larva, nymph and adult) was performed. For the determination of ticks, Leica Wild Stereo Microscope MZ8 (Leica Microsystems, Mannheim, Germany) was used (magnification 50×) equipped with an object micrometer together with the Quick Photo software package, ModellCamera 2 and a Dino-Lite digital microscope (magnification 20×–220×; 500×) (AnMo Electronics Corporation, Taiwan, China) with DinoCapture 2.0 software, version 1.5.17. B. Identification of sampled ticks based on different morphological characteristics was performed using current keys for tick identification [59,60]. After determining species and life stages, the ticks were stored in a freezer (−80 °C). The fieldwork was carried out under permits issued by the Ministry of Economy and Sustainable Development of the Republic of Croatia (UP/I-612-07/19-48/154, 517-05-1-1-19-3, 26 June 2019).

3.2. Borrelia burgdorferi s.l. Detection

Ticks were organized in pools (2–20 individuals per pool; median pool size = 11) in accordance with the locality, season (spring or autumn), altitude (200, 400, 600, 800 or 1000 m asl.) and tick life stage (nymph or adult; larvae were not included). When necessary, due to the size of the samples, pools included tick samples from two altitudes and both developmental stages. The study site of Medvednica included 18 pools with 165 nymphs and 69 adults; in Gorski Kotar, there were 54 nymphs and adults grouped together in 5 pools, while the study site of Papuk included 4 pools with 18 nymphs and adults combined. For specimens collected on Medvednica and Papuk, grouping according to altitudes was performed for ticks sampled at 200 and 400 m asl. and at 600 and 800 m asl. For ticks from Gorski Kotar, the same type of grouping was performed for specimens collected at 600 and 800 m asl.
Ticks were homogenized in a Qiagen tissuelyser II (Qiagen, Retsch, Germany) for 10 min at 30 Hz (using 1–3 mm corundum beads (Macherey-Nagel), 3 mm steel beads (Macherey-Nagel), 5 mm metal beads (IKA) and 500 μL of PBS). After homogenization, genomic DNA was extracted using a commercial kit (NucleoSpinⓇ Tissue kit, Macherey-Nagel, Düren, Germany) following the manufacturer’s instructions. B. burgdorferi s.l. DNA was detected by the nested PCR method targeting a specific ospA gene, as already described [61]. Nucleotide sequencing of the PCR products was performed by Macrogen Europe, Inc. (https://macrogenlab.com) (accessed on 9 January 2023). The resulting sequences were analyzed using BioEdit software v7.0 (4 https://bioedit.software.informer.com) (accessed on 9 January 2023). Phylogenetic analyses were conducted using MEGA version 11 (https://www.megasoftware.net) (accessed on 9 January 2023).

3.3. Statystical Analysis

The differences observed between the numbers of ticks collected at different altitudes were tested by Chi-square analysis. This was used to test the null hypothesis that the collected numbers of ticks would be similar for all altitudes. Data were analyzed using Microsoft Excel 365 and Statistica ver. 14.0.1.25., TIBCO Software, Inc. (Palo Alto, CA, USA).

4. Results

During this study (2019–2021), 2942 questing ticks were collected by the flag dragging method at three mountain sites in inland Croatia. The collected ticks belonged to three species: two were from the genera Ixodes and one was from Haemaphysalis (Table 3). Ixodes ricinus L., 1758, (Castor bean tick, Sheep tick, Deer tick, Forest tick) was by far the most dominant species (N = 2937; 99.83%), while Haemaphysalis concinna Koch, 1844 and Ixodes frontalis Panzer, 1798 represented less than 1% of the catch. Overall, 94.05% of all sampled specimens, including every tick species, were subadults. The largest number of collected Ixodes ricinus were in larval stage (53.90%), closely followed by nymphs (40.14%), while adults represented a little under 6% of the catch. Regarding the Ixodes ricinus sex ratio, males were slightly predominant (3.51%) over females (2.45%). Ixodes ricinus was the only tick species that was sampled at all life stages (larval, nymphal and adult) at every location, i.e., in the study area, while the other two species were collected only as subadults (Table 3).
The highest number of ticks was collected at the study site of Gorski Kotar (N = 1430; 48.6%), followed by Medvednica (N = 1129; 38.4%) and Papuk (N = 383; 13.0%). The list of tick species and their sex ratios and life stages according to study sites are shown in Table 4.
Ixodes ricinus was the one species found at all altitudes and sampling sites, while Haemaphysalis concinna was recorded only on Papuk (at 200 m asl. and 600 m asl.) and Ixodes frontalis only on Medvednica (200 m asl.) (Table 4).
The Chi-square analysis showed that altitude significantly influenced the number of collected ticks in all three mountain areas—Gorski Kotar (χ2 = 201.73, p < 0.05), Medvednica (χ2 = 442.95, p < 0.05) and Papuk (χ2 = 103.38, p < 0.05). However, sampling of the same altitudes (600 m asl and 800 m asl) on the mountains of Gorski Kotar and Medvednica resulted in similar numbers of collected ticks (Table 4). At these altitudes, there was no significant difference in the numbers of collected specimens (χ2 = 0.78, p > 0.05; χ2 = 0.88, p > 0.05).
Ticks were least numerous in their adult developmental stage in tick samples at all three study sites (Gorski Kotar: 1.96%; Medvednica: 12.13%; Papuk: 2.61%). The highest overall share of adults (33.33%) was recorded on Medvednica, at 400 m asl., in European beech forest with Woodruff (Table 4). Larvae dominated overall catches in Gorski Kotar (75.87%), while nymphs were the most dominant (60.23%) on Medvednica (Table 4). On Papuk, both larvae and nymphs had approximately equal overall shares (48%) in tick samples. The tick species, their sex ratios and life stages according to study sites and sampling seasons are shown in Table 5.
During the springtime sampling, 89.1% of all the ticks were collected. Nymphs were sampled both in spring and autumn at all mountain sites during the whole research period. Larvae were also sampled in spring and autumn at all sampling sites (Table 5).
Seasonal dynamics of ticks at study sites according to altitudes and forest communities are shown in Figure 2, Figure 3 and Figure 4. During every tick sampling, at all mountain sites, springtime was the season when ticks were collected in the highest numbers (Figure 2, Figure 3 and Figure 4). The exception from the usual springtime maximum was only recorded during 2021 at 400 m asl. and 600 m asl. sampling sites in Gorski Kotar (Figure 2), as well as at all altitudes (200, 400, 600 and 800 m asl.) on Papuk (Figure 4).
The highest density of ticks per 100 m2 recorded on Gorski Kotar (207 ticks per 100 m2) was at 800 m asl. within mixed Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992). At the same altitude, the highest tick density was also recorded on Medvednica (109 ticks per 100 m2) but within European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959), while on Papuk, at 200 m asl., in Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974), 42 ticks were collected per 100 m2 (Table 4).
The overall B. burgdorferi s.l. prevalence was 3.7%. In terms of sampling location, the presence of B. burgdorferi s.l. was detected in 20% (1/5) of pools containing nymphs and adults collected in the area of Gorski Kotar. B. burgdorferi s.l. infection was not detected in samples from the Medvednica and Papuk mountains. Sequencing of the ospA gene and phylogenetic analysis revealed the presence of the B. burgdorferi sensu stricto genotype. A phylogenetic tree, constructed using ospA sequences retrieved from GenBank and the sequence identified in this study (14N CRO), is shown in Appendix B (Figure A14).

5. Discussion

As the European Centre for Disease Prevention and Control (ECDC) predicts that the incidence of vector-borne diseases will continue to rise [8,62], it is no surprise that approximately three times as many publications on Ixodes ricinus have appeared since 2002 compared with the period between 1980 and 2000 [37]. Despite a large number of faunistic studies on the diversity of tick fauna conducted during the second half of the 20th century along the Adriatic Coast and a few recent studies in northwestern, central and eastern regions of inland Croatia, many areas—including mountains—have not yet been sufficiently studied. In this research, 3 out of 23 previously reported hard tick species (Ixodes ricinus, Ixodes frontalis and Haemaphysalis concinna) were collected in three mountain regions in inland Croatia (Gorski Kotar, Medvednica and Papuk) [48] (Figure 1). At all three mountain study sites, Ixodes ricinus was the most prevalent tick species (99.8% of all collected samples) (Table 3). These findings are in line with many studies performed earlier in woodland habitats at lower altitudes in northwestern and eastern parts of Croatia [20,44,46,63] as well as in Europe (e.g., 96% in Southern England, 97.4% in the Serbian capital Belgrade, 90.7% in Germany, etc.) [64,65,66,67,68]. Our results also differ from previous studies performed in pedunculate oak floodplain forests (Genisto elatae—Quercetum roboris Ht. 1938) in the Posavina region in Central Croatia [45], where Ixodes ricinus was determined in only 3.5% of collected specimens. On the mountain sites of Papuk and Medvednica, besides Ixodes ricinus, Ixodes frontalis and Haemaphysalis concinna were also recorded, but with very small percentages—0.08% and 1.0%, respectively (Table 4). Formerly, Ixodes frontalis was sporadically recorded in Croatia (Adriatic Coast and the city of Zagreb) [20], and this record for the Medvednica mountain represents a new locality record, as does the record of Haemaphysalis concinna on Papuk Mountain. There were also previous records of Haemaphysalis concinna sampled near the Croatian northwestern border, on the island of Krk in the Croatian Littoral [20] and in the eastern parts of the country, where it was present in small numbers, representing only 2.6% of the collected specimens [47]. Even though the genus Dermacentor is the second most abundant ixodid taxon in the Western Palaearctic region [69,70], we have collected no representatives in our three-year research, which corresponds to its lowland habitat preferences and its absence from higher mountain regions [45,71]. However, we expected ticks from this genus at low-altitude hill sampling sites or in climatically favorable valleys at higher altitudes [72].
In Central Europe, the altitudinal distribution limit for Ixodes ricinus was between 700 and 800 m asl. [73]. However, this limit has been exceeded, as shown by the results of the study on Krkonoše Mountain in the Czech Republic, where Ixodes ricinus was recorded even up to a timberline, approximately 1250 m asl. [74]. In Southern Norway, Ixodes ricinus was recorded at altitudes of up to 1000 m asl., which represents a considerable increase in altitude relative to earlier findings in this region [75]. In a similar study in Hungary, Ixodes ricinus was also found at altitudes of up to 900–1000 m asl. [76]. In this study, Ixodes ricinus was found at 1000 m asl. in Gorski Kotar and on the Medvednica mountain, these being the highest finding points of this species in Croatia.
The mean density of hard ticks in the study area was 18.9 per 100 m2. A significantly lower value was recorded for Ixodes ricinus in Northeastern Poland, namely, 9.7 ticks per 100 m2 [77]. Despite the different types of forest vegetation, the highest abundances and mean densities of ticks collected per 100 m2 were recorded at high altitudes (800–1000 m asl.) on Gorski Kotar and Medvednica and at lower altitudes (200 m asl.) on Papuk (Table 4), which matches this species’ habitat preferences (deciduous coniferous and mixed forests) [78,79,80,81,82,83]. The highest tick density (207 ticks per 100 m2) recorded in this study was at 800 m asl. on Gorski Kotar in mixed Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992). Such a high tick abundance is partly a result of the overall high prevalence of larvae (76%) collected on Gorski Kotar during this research (Table 4).
The seasonal dynamics of the sampled ticks showed a typical peak during springtime sampling [37], which corresponds to 89.1% of all the tick specimens collected during March, April and May in this study (Figure 2, Figure 3 and Figure 4). Except for a few exceptions, ticks were mostly sampled at all three developmental stages during every spring and autumn sampling (Table 5). Nymphs were sampled both in spring and in autumn at all mountain sites during the whole research period. Regarding seasonal dynamics, Ixodes ricinus larvae are known for their early summer peak [84], which partially corresponds to their dominance recorded during the springtime sampling in our study.
The only tick species found at all life stages (larval, nymphal and adult) at every location in this study was Ixodes ricinus, while Ixodes frontalis and Haemaphysalis concinna were sampled only as subadults (Table 3). Overall, larvae of Ixodes ricinus were mainly predominant (53.9%) but closely followed by nymphs (40.1%), while only 6% of the sampled specimens were adults. These results differ from the previous findings of Krčmar et al. (2019), who collected 67% of ticks at an adult stage in eastern woodland parts of the country [47]. At the same time, our data correspond to a high share of subadults (88%) sampled in a forest habitat within Croatia’s capital, the city of Zagreb [43]. In floodplain forests of the Posavina region in Central Croatia, only adult Ixodes ricinus were sampled from 2011 to 2013 [45]. On the Medvednica mountain, Ixodes ricinus nymphs were prevalent at all altitudinal limits (200, 400, 600, 800 and 1000 m asl.), while at the mountain site of Papuk, they were prevalent at 400 m asl. and from 600 to 800 m asl. These data are also partly in agreement with data from Krkonoše Mts., where all ticks collected above 850 m asl. were nymphs [74]. These data are not surprising, since the nymphal stage is the one most frequently recorded when ticks are sampled by the flagging method [74]. In contrast, larval stages were prevalent in collected samples at altitudes above 600 m in Gorski Kotar.
One of the aims of our study was to determine the prevalence of the Lyme disease causative agent among sampled ticks. Of 27 pools, containing 305 ticks, 1 pool (3.7%) was positive for Borrelia burgdorferi sensu lato infection. The detected prevalence is in line with a recent study on pathogens of zoonotic importance that revealed 2.2% infection with Borrelia burgdorferi s.l. within ticks collected in the Scottish Highlands [85], but it also differs from previous research conducted in urban forests in Croatia’s capital which showed high infection rates (55%) within tick populations [26]. If we consider our three research sites, the presence of the pathogen was detected in 20% of the samples from Gorski Kotar, while samples from Medvednica and Papuk showed no infection. Our data partly correspond to prevalences of Borrelia spp. within tick populations in urban woodland habitats recorded in Germany (24–34%), Italy (26–36%), Lithuania (25%), Poland (11–27%), Slovakia (6–10%), Switzerland (18%) and Serbia (23.6%) [10,86]. Our findings are also in line with meta-analysis data of Borrelia spp. prevalence in ticks in Europe (18.6% in adults and 10.1% in nymphs) [87].
Eight different genospecies of Borrelia burgdorferi, specific to different hosts, can be found throughout Europe [85]. Sequencing of the ospA gene and phylogenetic analysis revealed the presence of the B. burgdorferi sensu stricto genotype in ticks from the mountain area of Gorski Kotar. These findings cohere with previous reports of Borrelia afzelii, Borrelia garinii and also B. burgdorferi sensu stricto detected in Ixodes ricinus ticks collected in a Lyme borreliosis endemic region of Northern Croatia [27]. Further monitoring of tick fauna, their hosts and the pathogens they carry, along with ecological studies of their habitats, would surely provide vital information on rapidly changing natural and urban environments. Worrying trends of increasing contact between human populations and ticks could make tick surveillance, monitoring and control a high priority in the near future.

6. Conclusions

This is the first report on the altitudinal distributions of hard ticks (Acari: Ixodidae) in mountain regions of Croatia. Three tick species (Ixodes ricinus, Haemaphysalis concinna and Ixodes frontalis) were recorded in the study areas. In all three areas, Ixodes ricinus was the most abundant species.
Ixodes frontalis and Haemaphysalis concinna were recorded for the first time both on the Medvednica mountain and on the Papuk mountain.
The highest tick abundances were recorded at higher altitudes (800–1000 m asl.) in Gorski Kotar and on the Medvednica mountain, while on the Papuk mountain the highest abundance was recorded at a lower altitude (200 m asl).
Most ticks were collected during the spring season (March–June)—89.12%—while in autumn (September–December) only 10.88% of ticks were sampled.
The mean tick density of the study area was 18.9 ticks per 100 m2—Gorski Kotar: 29.8 (range: 0–207.5); Medvednica: 18.8 (range: 0–109.5); Papuk: 8.0 (range: 0–42.5).
The presence of B. burgdorferi s.l. was detected in 20% (1/5) of the tick samples from Gorski Kotar, while the samples from Medvednica and Papuk showed no infection.
Findings of Borrelia burgdorferi sensu stricto in nymphs and adults of Ixodes ricinus from Gorski Kotar indicate the risk of occurrence of Lyme borreliosis at altitudes from 600–800 m asl., where the causative agent of this emerging disease had not been registered before within tick populations in Croatia.

Author Contributions

Conceptualization, M.V. and S.K.; methodology, M.V., S.K., M.B., J.H., V.M.P. and L.B.; software, M.V., S.K., M.B., V.M.P. and L.B.; formal analysis, M.V., S.K., J.H. and V.M.P.; investigation, M.V., M.B. and L.B.; data curation, M.V., M.B. and L.B.; writing—original draft preparation, M.V. and S.K., writing—review and editing, S.K., M.V. and J.M., visualization, M.V., supervision, J.M.; project administration, M.V. and M.B.; funding acquisition, J.M. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Faculty of Forestry and Wood Technology, University of Zagreb.

Institutional Review Board Statement

The animal study protocol was approved by the Ethic Committee of the Faculty of Forestry and Wood Technology University of Zagreb (Approval Code: EP01-22/23 Approval Date: 19 January 2023.)

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Acknowledgments

The authors wish to thank Faculty of Forestry and Wood Technology University of Zagreb for all their support and also Croatian Meteorological and Hydrological Service for providing the data from meteorological stations at study sites Gorski Kotar, Medvednica and Papuk.

Conflicts of Interest

The authors declare no conflict of interest.

Appendix A. Tick Sampling Sites

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Figure A1. Sampling site 1. Mountain site: Gorski Kotar/altitude: 400 m asl./forest soil: Pseudogley on sloping terrain/forest community: EBDF: European beech forest with Deer fern (Blechno- fagetum sylvaticae Ht. 1950).
Figure A1. Sampling site 1. Mountain site: Gorski Kotar/altitude: 400 m asl./forest soil: Pseudogley on sloping terrain/forest community: EBDF: European beech forest with Deer fern (Blechno- fagetum sylvaticae Ht. 1950).
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Figure A2. Sampling site 2. Mountain site: Gorski Kotar/altitude: 600 m asl./forest soil: Dystric cambisol/forest community: EFDF: European fir forest with Deer fern (Blechno—Abietetum Ht. 1950).
Figure A2. Sampling site 2. Mountain site: Gorski Kotar/altitude: 600 m asl./forest soil: Dystric cambisol/forest community: EFDF: European fir forest with Deer fern (Blechno—Abietetum Ht. 1950).
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Figure A3. Sampling site 3. Mountain site: Gorski Kotar/altitude: 800 m asl./forest soil: Dystric cambisol/forest community: DBF: Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992).
Figure A3. Sampling site 3. Mountain site: Gorski Kotar/altitude: 800 m asl./forest soil: Dystric cambisol/forest community: DBF: Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992).
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Figure A4. Sampling site 4. Mountain site: Gorski Kotar/altitude: 1000 m asl./forest soil: Dystric cambisol/forest community: MPF: Mixed private forest stand with European beech (Fagus sylvatica L.), European silver fir (Abies alba Mill.), Sycamore (Acer pseudoplatanus L.), European ash (Fraxinus excelsior L.) and Common hazel (Coryilus avellana L.).
Figure A4. Sampling site 4. Mountain site: Gorski Kotar/altitude: 1000 m asl./forest soil: Dystric cambisol/forest community: MPF: Mixed private forest stand with European beech (Fagus sylvatica L.), European silver fir (Abies alba Mill.), Sycamore (Acer pseudoplatanus L.), European ash (Fraxinus excelsior L.) and Common hazel (Coryilus avellana L.).
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Figure A5. Sampling site 5. Mountain site: Medvednica/altitude: 200 m asl./forest soil: Dystric cambisol/forest community: POEH: Pedunculate oak forest with European hornbeam (Carpino betuli—Quercetum roboris typicum Rauš 1969).
Figure A5. Sampling site 5. Mountain site: Medvednica/altitude: 200 m asl./forest soil: Dystric cambisol/forest community: POEH: Pedunculate oak forest with European hornbeam (Carpino betuli—Quercetum roboris typicum Rauš 1969).
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Figure A6. Sampling site 6. Mountain site: Medvednica/altitude: 400 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
Figure A6. Sampling site 6. Mountain site: Medvednica/altitude: 400 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
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Figure A7. Sampling site 7. Mountain site: Medvednica/altitude: 600 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
Figure A7. Sampling site 7. Mountain site: Medvednica/altitude: 600 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
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Figure A8. Sampling site 8. Mountain site: Medvednica/altitude: 800 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
Figure A8. Sampling site 8. Mountain site: Medvednica/altitude: 800 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
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Figure A9. Sampling site 9. Mountain site: Medvednica/altitude: 800 m asl./forest soil: Dolomitic Cambisol/forest community: PBF: Pannonian beech–fir forest (Festuco drymeiae—Abietetum Vukelić et Baričević 2007).
Figure A9. Sampling site 9. Mountain site: Medvednica/altitude: 800 m asl./forest soil: Dolomitic Cambisol/forest community: PBF: Pannonian beech–fir forest (Festuco drymeiae—Abietetum Vukelić et Baričević 2007).
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Figure A10. Sampling site 10. Mountain site: Papuk/altitude: 200 m asl./forest soil: Luvisol/forest community: SOFD: Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974).
Figure A10. Sampling site 10. Mountain site: Papuk/altitude: 200 m asl./forest soil: Luvisol/forest community: SOFD: Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974).
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Figure A11. Sampling site 11. Mountain site: Papuk/altitude: 400 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
Figure A11. Sampling site 11. Mountain site: Papuk/altitude: 400 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
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Figure A12. Sampling site 12. Mountain site: Papuk/altitude: 600 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
Figure A12. Sampling site 12. Mountain site: Papuk/altitude: 600 m asl./forest soil: Dystric cambisol/forest community: EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959).
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Figure A13. Sampling site 13. Mountain site: Papuk/altitude: 800 m asl./forest soil: Dystric cambisol/forest community: MEBBLA: Mountain European beech forest with (Lamio orvalae—Fagetum sylvaticae Ht. 1938).
Figure A13. Sampling site 13. Mountain site: Papuk/altitude: 800 m asl./forest soil: Dystric cambisol/forest community: MEBBLA: Mountain European beech forest with (Lamio orvalae—Fagetum sylvaticae Ht. 1938).
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Appendix B

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Figure A14. A phylogenetic tree constructed using ospA sequences retrieved from GenBank (accession numbers are shown in the tree) and the sequence identified in this study (14N CRO). Evolutionary distances were calculated using the Maximum Likelihood method and Tamura–Nei model in MEGA 11 with 1000 bootstrap replications. The scale bar indicates the nucleotide substitutions per site.
Figure A14. A phylogenetic tree constructed using ospA sequences retrieved from GenBank (accession numbers are shown in the tree) and the sequence identified in this study (14N CRO). Evolutionary distances were calculated using the Maximum Likelihood method and Tamura–Nei model in MEGA 11 with 1000 bootstrap replications. The scale bar indicates the nucleotide substitutions per site.
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References

  1. Smith, T.; Kilbourne, F.L. Investigations into the nature, causation and prevention of Texas or southern cattle fever. Bull. Bur. Anim. Ind. 1893, 1, 177–304. [Google Scholar]
  2. Dutton, J.E.; Todd, J.L. The nature of tick fever in the eastern part of Congo Free State, with notes on the distribution and bionomics of the tick. Br. Med. J. 1905, 2, 1259–1260. [Google Scholar]
  3. Dennis, D.T.; Piesman, J.F. Overview of Tick-Borne Infections of Humans. In Tick-Borne Diseases of Humans; Goodman, J.L., Dennis, D.T., Sonenshine, D.E., Eds.; ASM Press: Washington, DC, USA, 2005; pp. 3–11. [Google Scholar]
  4. Jongejan, F.; Uilenberg, G. The global importance of ticks. Parasitology 2004, 129, S3–S14. [Google Scholar] [CrossRef] [PubMed]
  5. Smith, R.; Takkinen, J. Editorial team Collective. Lyme borreliosis: Europe-wide coordinated surveillance and action needed? Euro Surveill. 2006, 11, 2977. [Google Scholar] [CrossRef]
  6. Bacon, R.M.; Kugeler, K.J.; Mead, P.S. Surveillance for Lyme disease—United States, 1992–2006. MMWR Surveill. Summ. 2008, 57, SS-10. [Google Scholar]
  7. Mysterud, A.; Jore, S.; Østerås, O.; Viljugrein, H. Emergence of tick-borne diseases at northern latitudes in Europe: A comparative approach. Sci. Rep. 2017, 7, 16316. [Google Scholar] [CrossRef] [Green Version]
  8. Bajer, A.; Beck, A.; Beck, R.; Behnke, J.M.; Dwuznik-Szarek, D.; Eichenberger, R.M.; Farkas, R.; Fuehrer, H.P.; Heddergott, M.; Jokelainen, P.; et al. Babesiosis in Southern, Central and Northeastern Europe: An Emerging and Re-Emerging Tick-Borne Disease of Humans and Animals. Microorganisms 2022, 10, 945. [Google Scholar] [CrossRef]
  9. Medlock, J.M.; Hansford, K.M.; Bormane, A.; Derdakova, M.; Estrada-Peña, A.; George, J.C.; Golovljeva, I.; Jaenson, T.G.T.; Jensen, J.K.; Jensen, P.M.; et al. Driving forces for changes in geographical distribution of Ixodes ricinus ticks in Europe. Parasit. Vectors 2013, 6, 1. [Google Scholar] [CrossRef] [Green Version]
  10. Snegiriovaite, J.; Radzijevskaja, J.; Palauskas, A. A brief review: The prevalence of tick borne pathogens in urban and suburban areas. Biologia 2020, 66, 242–255. [Google Scholar] [CrossRef]
  11. Aguirre, A.A.; Tabor, G.M. Global factors driving emerging infectious diseases: Impact on wildlife populations. Ann. N. Y. Acad. Sci. 2008, 1149, 1–3. [Google Scholar] [CrossRef]
  12. Spielman, A. The Emergence of Lyme Disease and human Babesiosis in a changing environment. Ann. N. Y. Acad. Sci. 1994, 740, 146–156. [Google Scholar] [CrossRef] [PubMed]
  13. Gilbert, L.; Maffey, G.L.; Ramsay, S.L.; Hester, A.J. The effect of deer management on the abundance of Ixodes ricinus in Scotland. Ecol. Appl. 2012, 22, 658–667. [Google Scholar] [CrossRef]
  14. Piesman, J.; Gern, L. Lyme borreliosis in Europe and North America. Parasitology 2004, 129, 191–220. [Google Scholar] [CrossRef] [PubMed]
  15. Hall, J.L.; Alpers, K.; Bown, K.J.; Martin, S.J.; Birtles, R.J. Use of mass-participation outdoor events to assess human exposure to tickborne pathogens. Emerg. Infect. Dis. 2017, 23, 463–467. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. Mrljak, V.; Kuleš, J.; Mihaljević, Ž.; Torti, M.; Gotić, J.; Crnogaj, M.; Živičnjak, T.; Mayer, I.; Šmit, I.; Bhide, M.; et al. Prevalence and geographic distribution of vector-borne pathogens in apparently healthy dogs in Croatia. Vector Borne Zoonotic Dis. 2017, 17, 398–408. [Google Scholar] [CrossRef]
  17. Estrada-Peña, A. Distribution, abundance, and habitat preferences of Ixodes ricinus (Acari: Ixodidae) in northern Spain. J. Med. Entomol. 2001, 38, 361–370. [Google Scholar] [CrossRef]
  18. Walker, A.R.; Alberdi, M.P.; Urquhart, K.A.; Rose, H. Risk factors in habitats of the tick Ixodes ricinus influencing human exposure to Ehrlichia phagocytophila bacteria. Med. Vet. Entomol. 2001, 15, 40–49. [Google Scholar] [CrossRef]
  19. Hansford, K.M.; Fonville, M.; Gillingham, E.L.; Copian, E.C.; Pietzch, M.E.; Krawczyk, A.I.; Vaux, A.G.C.; Cull, B.; Sprong, H.; Medlock, J.M. Ticks and Borrelia in urban and peri-urban green space habitats in a city in southern England. Ticks Tick Borne Dis. 2017, 8, 353–361. [Google Scholar] [CrossRef]
  20. Krčmar, S. Hard ticks (Acari, Ixodidae) of Croatia. ZooKeys 2012, 234, 19–57. [Google Scholar] [CrossRef] [Green Version]
  21. Gray, J.S. Biology of Ixodes species ticks in relation to tick-borne zoonoses. Wien. Klin. Wochenschr. 2002, 114, 473–478. [Google Scholar]
  22. García-Álvarez, L.; Palomar, A.M.; Oteo, J.A. Prevention and prophylaxis of tick-bites and tick-borne related diseases. Am. J. Infect. Dis. 2013, 9, 104–116. [Google Scholar] [CrossRef] [Green Version]
  23. Mulić, R.; Petković, B.; Klišmanić, Z.; Jerončić, I. Tick-borne diseases in the Republic of Croatia. Lječ. Vjesn. 2011, 133, 89–95. [Google Scholar]
  24. Beck, R.; Habrun, B.; Bosnić, S.; Benić, M.; Nemeth-Blažić, T.; Duvnjak, S. Identification of pathogens in Ixodes ricinus and Dermacentor reticulatus fron public gardens in Zagreb, Croatia. In Proceedings of the 12th International Conference on Lyme Borreliosis and Other Tick-Borne Diseases, Ljubljana, Slovenia, 26–29 September 2010. [Google Scholar]
  25. Golubić, D.; Rijpkema, S.; Tkalec-Makovec, N.; Ruzić, E. Epidemiologic, ecologic and clinical characteristics of Lyme borreliosis in northwest Croatia. Acta Med. Croat. 1998, 52, 7–13. [Google Scholar]
  26. Barišin, A.; Nemeth Blažić, T.; Jeličić, P.; Gjenero Margan, I.; Capak, K.; Petrović, G. Prikaz istraživanja krpelja na području Grada Zagreba u 2008. godini. Zbornik radova DDD-ZUPP 2011, 1, 203–211. [Google Scholar]
  27. Rijepkema, S.; Golubić, D.; Molkenboer, M.; Verbeek, D.E.; Kruif, N.; Schellekens, J.F.P. Identification of four genomic groups of B. burgdorferi sensu lato in Ixodes ricinus ticks collected in a Lyme borreliosis endemic region on northern Croatia. Exp. Appl. Acarol. 1996, 20, 25–30. [Google Scholar]
  28. Steere, A.C.; Malawista, S.E.; Snydman, D.R.; Shope, R.E.; Andiman, W.A.; Ross, M.R.; Steele, F.M. Lyme arthritis: An epidemic of oligoarticular arthritis in children and adults in three connecticut communities. Arthritis Rheum. 1977, 20, 7–17. [Google Scholar] [CrossRef]
  29. Parola, F.; Raoult, D. Tick and tickborne bacterial diseases in humans: An emerging infectious threat. Ticks Tick Borne Dis. 2001, 32, 897–928. [Google Scholar] [CrossRef] [Green Version]
  30. Quine, C.P.; Barnett, J.; Dobson, A.D.M.; Marcu, A.; Marzano, M.; Moseley, D.; O’ Brien, L.; Randolph, S.E.; Taylor, J.L.; Uzell, D. Frameworks for risk communication and disease management: The case of Lyme disease and countryside users. Philos. Trans. R. Soc. B Biol. Sci. 2011, 366, 2010–2022. [Google Scholar] [CrossRef] [PubMed]
  31. Rizzoli, A.; Hauffe, H.C.; Carpi, G.; Vourc’h, G.I.; Neteler, M.; Rosà, R. Lyme borreliosis in Europe. Euro Surveill. 2011, 16, 19906. [Google Scholar] [CrossRef] [PubMed]
  32. Strnad, M.; Hönig, V.; Růžek, D.; Grubhoffer, L.; Rego, R.O.M. Europe-wide meta-analysis of Borrelia burgdorferi sensu lato prevalence in questing Ixodes ricinus ticks. Appl. Environ. Microbiol. 2017, 83, e00609-17. [Google Scholar] [CrossRef] [Green Version]
  33. Bennet, L.; Halling, A.; Berglund, J. Increased incidence of Lyme borreliosis in southern Sweden following mild winters and during warm, humid summers. Eur. J. Clin. Microbiol. Infect. Dis. 2006, 25, 426–432. [Google Scholar] [CrossRef] [Green Version]
  34. Croatian Institute of Public Health. Statistical Yearbooks (1996–2021). Available online: https://www.hzjz.hr/hrvatski-zdravstveno-statisticki-ljetopis/hrvatski-zdravstveno-statisticki-ljetopis-za-2020-tablicni-podaci/ (accessed on 2 January 2023).
  35. Lipozenčić, J.; Šitum, M. Epidemiology of Lyme disease. Acta Derm. APA 2001, 10, 1–9. [Google Scholar]
  36. Estrada-Peña, A.; Cutler, S.; Potkonjak, A.; Vassier-Tussaut, M.; Van Bortel, W.; Zeller, H.; Fernández-Ruiz, N.; Mihalca, A.D. An updated meta-analysis of the distribution and prevalence of Borrelia burgdorferi s.l. in ticks in Europe. Int. J. Health Geogr. 2018, 17, 41. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  37. Kahl, O.; Gray, J.S. The biology of Ixodes ricinus with emphasis on its ecology. Ticks Tick Borne Dis. 2023, 14, 102114. [Google Scholar] [CrossRef] [PubMed]
  38. Mikačić, D. The ticks of the littoral zone in Yugoslavia. I. The morphology of the species from the genus Hyalomma. Vet. Arhiv. 1961, 31, 305–310. [Google Scholar]
  39. Mikačić, D. The ticks of the littoral zone of Yugoslavia. II. The genus Haemaphysalis with reference to the distinction between H. punctata and H. cholodkovskyi. Vet. Arhiv. 1963, 33, 133–136. [Google Scholar]
  40. Mikačić, D. Ticks in the littoral belt of Yugoslavia III. Distribution and dynamics of species in the course of the year. Vet. Arhiv. 1965, 35, 155–170. [Google Scholar]
  41. Tovornik, D.; Brelih, S. Ixodid ticks, the parasites of lizards (Lacertidae) in the karst and other districts of Yugoslavia. Scopolia 1980, 3, 1–21. [Google Scholar]
  42. Tovornik, D.; Vesenjak-Hirjan, J. A revision of ticks belonging to the Rhipicephalus sanguineus complex (Latreille), collected in the Yugoslav Coastal Region. Biol. Vestn. 1988, 36, 77–84. [Google Scholar]
  43. Vucelja, M.; Bjedov, L.; Boljfetić, M.; Klobučar, A.; Krčmar, S.; Borak, S.; Modrić, M.; Juričić, K.; Peleš, V.; Margaletić, J.; et al. Monitoring of Hard Ticks at Urban Recreational Sites in the City of Zagreb from 2016 to 2018. Infektol. Glasn. 2019, 39, 33–39. [Google Scholar] [CrossRef]
  44. Mikačić, D. Dinamika pojavljivanja krpelja (Ixodidae) u Sjevernoj Hrvatskoj. Vet. Arhiv. 1969, 39, 183–186. [Google Scholar]
  45. Vucelja, M. Rodent Control (Rodentia, Murinae, Arvicolinae) in Pedunculate Oak Forests (Quercus robur L.)—Integrated Approach and Zoonotic Aspect, 1st ed.; University of Zagreb, Faculty of forestry: Zagreb, Croatia, 2013; pp. 25–210. [Google Scholar]
  46. Borčić, B.; Aleraj, B.; Žutić, M.; Mikačić, D. The role of ticks (Ixodidae) in the maintenance of the tularemia natural focus in central Posavinia. Vet. Arhiv. 1978, 48, 277–283. [Google Scholar]
  47. Krčmar, S. Diversity, ecology, and seasonality of hard ticks (Acari: Ixodidae) in eastern Croatia. J. Vector Ecol. 2019, 44, 18–29. [Google Scholar] [CrossRef] [Green Version]
  48. Krčmar, S.; Klobučar, A.; Vucelja, M.; Boljfetić, M.; Kučinić, M.; Madić, J.; Cvek, M.; Bruvo Mađarić, B. DNA barcoding of hard ticks (Ixodidae), notes on distribution of vector species and new faunal record for Croatia. Ticks Tick Borne Dis. 2022, 13, 101920. [Google Scholar] [CrossRef]
  49. EEA. European Environment Agency Biogeographic Regions Dataset. 2016. Permalink: 2a7b433b388e432d9c3e8cf019af1db0. Available online: https://www.eea.europa.eu/data-and-maps/data/biogeographical-regions-europe-3 (accessed on 8 January 2023).
  50. Filipčić, A. Klimatska regionalizacija Hrvatske po Köppenu za standardno razdoblje 1961–1990. u odnosu na razdoblje 1931–1960. Acta Geograph. Croat. 1998, 34, 1–15. [Google Scholar]
  51. Zaninović, K.; Gajić-Čapka, M.; Perčec Tadić, M.; Vučetić, M.; Milković, J.; Bajić, A.; Cindrić, K.; Cvitan, L.; Katušin, Z.; Kaučić, D.; et al. Klimatski atlas Hrvatske/Climate atlas of Croatia 1961–1990, 1971–2000, 1st ed.; Državni Hidrometeorološki Zavod Zagreb: Zagreb, Hrvatska, 2008; pp. 1–200. [Google Scholar]
  52. Šegota, T.; Filipčić, A. Köppenova podjela klima i hrvatsko nazivlje. Geoadria 2003, 8, 17–31. [Google Scholar] [CrossRef] [Green Version]
  53. Vukelić, J.; Rauš, Đ. Forest phytocenology and forest communities in Croatia (Šumarska fitocenologija i šumske zajednice u Hrvatskoj), 1st ed.; University of Zagreb, Faculty of forestry: Zagreb, Croatia, 1988; pp. 173–177. [Google Scholar]
  54. Vukelić, J. Croatian Forest Vegetation (Šumska Vegetacija Hrvatske), 1st ed.; University of Zagreb, Faculty of forestry: Zagreb, Croatia, 2012; pp. 81–101. [Google Scholar]
  55. Martinović, J. Management of Forest Soils in Croatia, 1st ed.; Gračan, J., Ed.; Croatian Forests Limited Liability Company, Ministry of Science and technology: Zagreb, Croatia, 2003; pp. 118–119. [Google Scholar]
  56. Cohnstaedt, L.W.; Rochon, K.; Duehl, A.J.; Anderson, J.F.; Barrera, R.; Yao Su, N.; Gerry, A.C.; Obenauer, P.J.; Campbell, J.F.; Lysyk, T.J.; et al. Arthropod Surveillance Programs: Basic Components, Strategies, and Analysis. Ann. Entomol. Soc. Am. 2012, 105, 135–149. [Google Scholar] [CrossRef] [Green Version]
  57. Falco, R.; Fish, D. The comparison of methods for sampling the deer tick, Ixodes dammini, in a Lyme disease endemic area. Exp. Appl. Acarol. 1992, 14, 165–173. [Google Scholar] [CrossRef]
  58. Carroll, J.F.; Schmidtmann, E.T. Tick sweep: Modification of the tick drag—Flag method for sampling nymphs of the deer tick (Acari: Ixodidae). J. Med. Entomol. 1992, 29, 352–355. [Google Scholar] [CrossRef] [PubMed]
  59. Estrada-Peña, A.; Bouattour, A.; Camicas, J.L.; Walker, A.R. Ticks of Domestic Animals in the Mediterranean Region—A Guide to Identification of Species, 1st ed.; University of Zaragoza: Zaragoza, Spain, 2004; pp. 1–128. [Google Scholar]
  60. Estrada-Peña, A.; Mihalca, A.D.; Petney, T.N. Ticks of Europe and North Africa. A Guide to Species Identification, 1st ed.; Springer International Publishing AG: Cham, Szwitzerland, 2017; pp. 1–404. [Google Scholar]
  61. Guy, E.C.; Stanek, G. Detection of Borrelia burgdorferi in patients with Lyme disease by the polymerase chain reaction. J. Clin. Pathol. 1991, 44, 610–611. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  62. Lindgren, E.; Andersson, Y.; Suk, J.E.; Sudre, B.; Semenza, J.C. Public health. Monitoring EU emerging infectious disease risk due to climate change. Science 2012, 336, 418–419. [Google Scholar] [CrossRef] [Green Version]
  63. Kovačević, J.; Bučanović, T.; Krčmar, S. Hard tick fauna (Acari: Ixodidae) in different types of habitats in the city of Osijek (Eastern Croatia). Nat. Croat. 2020, 29, 63–72. [Google Scholar] [CrossRef]
  64. Cull, B.; Vaux, A.G.C.; Ottowell, L.J.; Gillingham, E.L.; Medlock, J.M. Tick infestation of small mammals in an English woodland. J. Vector Ecol. 2017, 42, 74–83. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  65. Milutinović, M.; Radulović, Ž.; Tomanović, S.; Tomanović, Ž. Seasonal distribution of Borreliae in Ixodes ricinus ticks in the Belgrade region, Serbia. Arch. Biol. Sci. Belgrade 2006, 58, 183–186. [Google Scholar] [CrossRef]
  66. Estrada-Peña, A.; Venzal, J.M.; Sánchez Acedo, C. The tick Ixodes ricinus: Distribution and climate preferences in the western Palaearctic. Med. Vet. Entomol. 2006, 20, 189–197. [Google Scholar] [CrossRef] [PubMed]
  67. Accrosi, A.; Schiavetti, I.; Listorti, V.; Dellepiane, M.; Masotti, C.; Ercolini, C.; Guardone, L.; Razzuoli, E. Hard ticks (Ixodidae) from Wildlife in Liguria, Northwest Italy: Tick Species Diversity and Tick-Host Associations. Insects 2022, 13, 199. [Google Scholar] [CrossRef]
  68. Bertola, M.; Momtarsi, F.; Obber, F.; Da Rold, G.; Carlin, S.; Toniolo, F.; Porcellato, E.; Falcaro, C.; Mondardini, V.; Ormelli, S.; et al. Occurrence and identification of Ixodes ricinus borne pathogens in Northeastern Italy. Pathogens 2021, 10, 1181. [Google Scholar] [CrossRef]
  69. Rubel, F.; Brugger, K.; Pfeiffer, M.; Chitimia-Dobler, I.; Didyk, Y.M.; Leverenz, S.; Dautel, H.; Kahl, O. Geographical distribution of Dermacentor marginatus and Dermacentor reticulatus in Europe. Ticks Tick Borne Dis. 2016, 7, 224–233. [Google Scholar] [CrossRef] [Green Version]
  70. Bilbija, B.; Spitzweg, C.; Papoušek, I.; Fritz, U.; Földvári, G.; Mullett, M.; Ihlow, F.; Sprong, H.; Civáňová Krížová, K.; Anisimov, N.; et al. Dermacentor reticulatus—A tick on its way from glacial refugia to a panmictic Eurasian population. Int. J. Parasitol. 2023, 53, 91–101. [Google Scholar] [CrossRef]
  71. Široký, P.; Kubelová, M.; Bednář, M.; Modrý, D.; Hubálek, Z.; Tkadlec, E. The distribution and spreading pattern of Dermacentor reticulatus over its threshold area in the Czech Republic—How much is range of this vector expanding? Vet. Parasitol. 2011, 183, 130–135. [Google Scholar] [CrossRef]
  72. Földvári, G.; Široký, P.; Szekeres, S.; Majoros, G.; Sprong, H. Dermacentor reticulatus: A vector on the rise. Parasit. Vectors 2016, 9, 314. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  73. Daniel, M. Influence of the microclimate on the vertical distribution of the tick Ixodes ricinus (L.) in Central Europe. Acarologia 1993, 34, 105–113. [Google Scholar]
  74. Materna, J.; Daniel, M.; Danielova, V. Altitudinal distribution limit of the tick Ixodes ricinus shifted considerably towards higher altitudes in central Europe: Results of thre years monitoring in the Krkonoše Mts. (Czech Republic). Cent. Eur. J. Publ. Health 2005, 13, 24–28. [Google Scholar]
  75. Pelsmaeker, N.D.; Korslund, L.; Steifetten, Ø. High-elevational occurrence of two tick species, Ixodes ricinus and I. trianguliceps, at their northern distribution range. Parasit. Vectors 2021, 14, 161. [Google Scholar] [CrossRef]
  76. Hornok, S.; Farkas, R. Influence of biotope on the distribution and peak activity of questing ixodid ticks in Hungary. Med. Vet. Entomol. 2009, 23, 41–46. [Google Scholar] [CrossRef] [PubMed]
  77. Kubiak, K.; Szymaňska, H.; Dmitryjuk, M.; Dzika, E. Abundance of Ixodes ricinus ticks (Acari: Ixodidae) and the diversity of Borrelia species in Northeastern Poland. Int. J. Environ. Res. Public Health 2022, 19, 7378. [Google Scholar] [CrossRef] [PubMed]
  78. Otranto, D.; Dantas-Torres, F.; Santos-Silva, M.M. Ixodes ricinus (Linnaeus, 1758) (Figs.67–69). In Ticks of Europe and North Africa: A Guide to Species Identification, 1st ed.; Estrada-Peña, A., Mihalca, A.D., Petney, T.N., Eds.; Springer International Publishing AG: Cham, Szwitzerland, 2017; pp. 189–195. [Google Scholar]
  79. Paul, R.E.L.; Cote, M.; Le Naour, E.; Bonnet, S.I. Environmental factors influencing tick densities over seven years in a French suburban forest. Parasit. Vectors 2016, 9, 309. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  80. Estrada-Peña, A.; De La Fuente, J. Species interactions in occurrence data for a community of tick transmitted pathogens. Sci. Data 2016, 3, 160056. [Google Scholar] [CrossRef] [Green Version]
  81. Schwarz, A.; Maier, W.A.; Kistemann, T.; Kampen, H. Analysis of the distribution of the tick Ixodes ricinus L. (Acari: Ixodidae) in a nature reserve of western Germany using geographic information systems. Int. J. Hyg. Environ. Health 2009, 212, 87–96. [Google Scholar] [CrossRef]
  82. Gray, J.S.; Dautel, H.; Estrada-Peña, A.; Kahl, O.; Lindgren, E. Effects of Climate Change on Ticks and Tick-Borne Diseases in Europe. Inter. Perspect. Infect. Dis. 2009, 212, 87–96. [Google Scholar] [CrossRef]
  83. Ehrmann, S.; Liira, J.; Gärtner, S.; Hansen, K.; Brunet, J.; Cousins, S.A.O.; Deconchat, M.; Decocq, G.; De Frenne, P.; De Smedt, P.; et al. Environmental drivers of Ixodes ricinus abundance in forest fragments of rural European landscapes. BMC Ecol. 2017, 17, 31. [Google Scholar] [CrossRef] [PubMed]
  84. Petney, T.N.; Pfäffie, M.P.; Skuballa, J.D. An annotated checklist of the ticks (Acari: Ixodidae) of Germany. Syst. Appl. Acarol. 2012, 17, 115–170. [Google Scholar] [CrossRef]
  85. Olsthoorn, F. The Effect of Woodland Expansion on Tick Populations and Lyme Disease Risk. Ph.D. Thesis, ETH Zürich, Zürich, Switzerland, 2021. [Google Scholar]
  86. Milutinović, M.; Radulović, Ž.; Jovičić, V.; Oreščanin, Z. Population dynamics and Borrelia burgdorferi infection rate of Ixodes ricinus ticks in the Belgrade area. Acta Vet. 2004, 54, 219–225. [Google Scholar]
  87. Rauter, C.; Hartung, T. Prevalence of Borrelia burgdorferi sensu lato genospecies in Ixodes ricinus ticks in Europe: A metaanalysis. Appl. Environ. Microbiol. 2005, 71, 7203–7216. [Google Scholar] [CrossRef] [PubMed] [Green Version]
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Figure 1. Study areas and locations of tick sampling sites. (Gorski kotar mountain site: 1—Donje Tihovo, 2—Marija Trošt, 3—Lučice, 4—Stari Laz; Medvednica mountain site: 5 —Dotršćina, 6–9 Medvednica a,b,c,d; Papuk mountain site: 10—Markovac, 11, 12—Slatinski Drenovac a,b, 13—Radovanci).
Figure 1. Study areas and locations of tick sampling sites. (Gorski kotar mountain site: 1—Donje Tihovo, 2—Marija Trošt, 3—Lučice, 4—Stari Laz; Medvednica mountain site: 5 —Dotršćina, 6–9 Medvednica a,b,c,d; Papuk mountain site: 10—Markovac, 11, 12—Slatinski Drenovac a,b, 13—Radovanci).
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Figure 2. Seasonal dynamics of ticks (Ixodes ricinus) sampled at the Gorski Kotar study site (2019–2021) for different altitudes and forest communities (S: spring, A: autumn); EFDF: European fir forest with Deer fern (Blechno—Abietetum Ht. 1950); DBF: Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992); MPF: Mixed private forest stand with European beech (Fagus sylvatica L.), European silver fir (Abies alba Mill.), Sycamore (Acer pseudoplatanus L.), European ash (Fraxinus excelsior L.) and Common hazel (Coryilus avellana L.).
Figure 2. Seasonal dynamics of ticks (Ixodes ricinus) sampled at the Gorski Kotar study site (2019–2021) for different altitudes and forest communities (S: spring, A: autumn); EFDF: European fir forest with Deer fern (Blechno—Abietetum Ht. 1950); DBF: Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992); MPF: Mixed private forest stand with European beech (Fagus sylvatica L.), European silver fir (Abies alba Mill.), Sycamore (Acer pseudoplatanus L.), European ash (Fraxinus excelsior L.) and Common hazel (Coryilus avellana L.).
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Figure 3. Seasonal dynamics of ticks sampled at the Medvednica study site (2019–2021) for different altitudes and forest communities (S: spring, A: autumn); POEH: Pedunculate oak forest with European hornbeam (Carpino betuli—Quercetum roboris typicum Rauš 1969); EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959); PBF: Pannonian beech–fir forest (Festuco drymeiae—Abietetum Vukelić et Baričević 2007).
Figure 3. Seasonal dynamics of ticks sampled at the Medvednica study site (2019–2021) for different altitudes and forest communities (S: spring, A: autumn); POEH: Pedunculate oak forest with European hornbeam (Carpino betuli—Quercetum roboris typicum Rauš 1969); EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959); PBF: Pannonian beech–fir forest (Festuco drymeiae—Abietetum Vukelić et Baričević 2007).
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Figure 4. Seasonal dynamics of ticks sampled at the Papuk study site (2019–2021) for different altitudes and forest communities (S: spring, A: autumn); SOFD: Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974); MEBBLA: Mountain European beech forest with balm-leaved archangel (Lamio orvalae—Fagetum sylvaticae Ht. 1938).
Figure 4. Seasonal dynamics of ticks sampled at the Papuk study site (2019–2021) for different altitudes and forest communities (S: spring, A: autumn); SOFD: Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974); MEBBLA: Mountain European beech forest with balm-leaved archangel (Lamio orvalae—Fagetum sylvaticae Ht. 1938).
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Table
Table 1. List of tick sampling sites with their GPS positions, altitudes and forest soil and forest community types.
Table 1. List of tick sampling sites with their GPS positions, altitudes and forest soil and forest community types.
Mountain SiteTick Sampling
Site		Altitude, Latitude
(Degree/min/s)		Altitude
(m asl.)		Forest
Soil		Forest
Community
GOK1 Donje TihovoN45 25.927 E14 50.814;400Pseudogley on sloping terrainEBDF
2 Marija TroštN45 24.798 E14 49.351;600Dystric cambisolEFDF
3 LučiceN45 22.359 E14 48.402;800Dystric cambisolDBF
4 Stari LazN45 20.740 E14 51.486;1000Dystric cambisolMPF
MED5 DotrščinaN45 51.174 E16 01.172;200Dystric cambisolPOEH
6 Medvednica aN45 52.748 E15 58.568:400Dystric cambisolEBW
7 Medvednica bN45 53.201 E15 58.330;600Dystric cambisolEBW
8 Medvednica cN45 53.394 E15 57.634;800Dystric cambisolEBW
9 Medvednica dN45 53.916 E15 56.753;1000Dolomitic cambisolPBF
PAP10 MarkovacN45 25.583 E17 32.227;200LuvisolSOFD
11 Slatinski Drenovac aN45 31.553 E17 41.306;400Dystric cambisolEBW
12 Slatinski Drenovac bN45 30.850 E17 39.990600Dystric cambisolEBW
13 RadovanciN45 30.673 E17 39.430;800Dystric cambisolMEBBLA
Legend: GOK: Gorski Kotar; MED: Medvednica; PAP: Papuk; EBDF: European beech forest with Deer fern (Blechno- fagetum sylvaticae Ht. 1950); EFDF: European fir forest with Deer fern (Blechno—Abietetum Ht. 1950); DBF: Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992); MPF: Mixed private forest stand with European beech (Fagus sylvatica L.), European silver fir (Abies alba Mill.), Sycamore (Acer pseudoplatanus L.), European ash (Fraxinus excelsior L.) and Common hazel (Coryilus avellana L.); POEH: Pedunculate oak forest with European hornbeam (Carpino betuli—Quercetum roboris typicum Rauš 1969); EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959); PBF: Pannonian beech–fir forest (Festuco drymeiae—Abietetum Vukelić et Baričević 2007); SOFD: Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974); MEBBLA: Mountain European beech forest with balm-leaved archangel (Lamio orvalae—Fagetum sylvaticae Ht. 1938).
Table
Table 2. Mean air temperatures and air humidity data for spring and autumn tick sampling seasons at three mountain sites.
Table 2. Mean air temperatures and air humidity data for spring and autumn tick sampling seasons at three mountain sites.
GOK/Air Temperature (°C) (min–max)GOK/Air Humidity (%) (min–max)
Year
Season		201920202021201920202021
S10.7 (0.2–14.9)10.2 (−4.5–22.2)9.9 (−3.9–25.3)82 (57–97)78.5 (45–97)85.8 (71–98)
A8.5 (−4.1–18.6)7.4 (−4.8–18.0)6.7 (−3.7–18.8)89.3 (78–98)89.8 (75–98)90 (76–98)
MED/air temperature (°C) (min–max)MED/air humidity (%) (min–max)
S9.73 (−1.0–24.4)9.01 (−5.9–21.2)8.25 (−5.6–23.6)74.5 (36–100)67.5 (29–99)72.5 (31–100)
A7.53 (−6.6–21.3)6.63 (−6.4–18.4)5.9 (−5.4–19.0)82.5 (45–100)67.1 (48–100)84.5 (32–100)
PAP/air temperature (°C) (min–max)PAP/air humidity (%) (min–max)
S12.3 (0.2–24.6)11.3 (−3.0–22.7)10.8 (−2.0–26.4)76.3 (67–98)82.5 (59–96.6)81.4 (63–97.3)
A8.95 (−4–21.1)8.1 (−2.6–19.4)7.2 (−2.7–19.6)88.3 (77–98)88.8 (70.2–96.2)88.5 (73.6–97.1)
Legend: GOK: Gorski Kotar; MED: Medvednica; PAP: Papuk; S: spring; A: autumn.
Table
Table 3. Species identifications, sex ratios and developmental stages of hard ticks collected in the study areas.
Table 3. Species identifications, sex ratios and developmental stages of hard ticks collected in the study areas.
SpeciesFemalesMalesNymphsLarvaeTotal
Ixodes ricinus72103117915832937
Ixodes frontalis00101
Haemaphysalis concinna00314
Ʃ72103118315842942
Table
Table 4. List of collected tick species and their life stages according to sampling location, altitude and forest community type.
Table 4. List of collected tick species and their life stages according to sampling location, altitude and forest community type.
Mountain SiteAltitude
(m asl.)		Forest
Community		Ir ♀Ir ♂Ir nIr lIf nHc nHc lƩMean Density per 100 m2 (min–max)
GOK400EBDF0211625000036830.7 (1–131.5)
600EFDF54844200013511.3 (0.5–32.5)
800DBF646638300045938.3 (0–207.5)
1000MPF255141000046839 (0–177.5)
Ʃ 13153171085000143029.8
MED200POEH353438100816.75 (0–21.5)
400EBW2529941400016213.5 (0–22.5)
600EBW4141072500015012.5 (0.5–20.5)
800EBW81823722500048840.7 (13.5–109.5)
1000PBF1516208900024820.7 (1.5–63)
Ʃ 5582680311100112918.8
PAP200SOFD33897803017614.7 (0–42.5)
400EBW122910000423.5 (0–11.5)
600EBW002369001937.8 (0–38)
800MEBBLA014130000726 (0–23.5)
Ʃ 461821870313838.0
Ʃ Ʃ 7210311791583131294218.9
Legend: GOK: Gorski Kotar; MED: Medvednica; PAP: Papuk; Ir: Ixodes ricinus; If: Ixodes frontalis; Hc: Haemaphysalis concinna; ♀ (female), ♂ (male), n (nymph), l (larva); EBDF: European beech forest with Deer fern (Blechno- fagetum sylvaticae Ht. 1950); EFDF: European fir forest with Deer fern (Blechno—Abietetum Ht. 1950); DBF: Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992); MPF: Mixed private forest stand with European beech (Fagus sylvatica L.), European silver fir (Abies alba Mill.), Sycamore (Acer pseudoplatanus L.), European ash (Fraxinus excelsior L.) and Common hazel (Coryilus avellana L.); POEH: Pedunculate oak forest with European hornbeam (Carpino betuli—Quercetum roboris typicum Rauš 1969); EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959); PBF: Pannonian beech–fir forest (Festuco drymeiae—Abietetum Vukelić et Baričević 2007); SOFD: Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974); MEBBLA: Mountain European beech forest with balm-leaved archangel (Lamio orvalae—Fagetum sylvaticae Ht. 1938).
Table
Table 5. List of collected tick species and their life stages according to sampling location, altitude, forest community type and sampling season.
Table 5. List of collected tick species and their life stages according to sampling location, altitude, forest community type and sampling season.
Mountain SiteAltitude
(m asl.)		Forest
Community		Sp.201920202021
SASASA
GOK400EBDFIr0 ♀, 0 ♂, 26 n, 22 l0 ♀, 0 ♂, 2 n, 0 l0 ♀, 0 ♂, 41 n, 222 l0 ♀, 1 ♂, 26 n, 0 l0 ♀, 1 ♂, 4 n, 0 l0 ♀, 0 ♂, 17 n, 6 l
600EFDFIr0 ♀, 2 ♂, 16 n, 10 l0 ♀, 0 ♂, 1 n, 0 l4 ♀, 2 ♂, 38 n, 21 l1 ♀, 0 ♂, 12 n, 5 l0 ♀, 0 ♂, 4 n, 0 l0 ♀, 0 ♂, 13 n, 6 l
800DBFIr1 ♀, 0 ♂, 24 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 2 ♂, 30 n, 383 l0 ♀, 0 ♂, 0 n, 0 l2 ♀, 1 ♂, 5 n, 0 l2 ♀, 0 ♂, 1 n, 0 l
1000MPFIr1 ♀, 3 ♂, 21 n, 330 l0 ♀, 0 ♂, 0 n, 0 l1♀, 1 ♂, 20 n, 62 l0 ♀, 0 ♂, 6 n, 18 l0 ♀, 1 ♂, 4 n, 0 l2 ♀, 5 ♂, 51 n, 410 l
MED200POEHIr2 ♀, 3 ♂, 15 n, 3 l0 ♀, 1 ♂, 2 n, 7 l1♀, 1 ♂, 13 n, 28 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 4 n, 0 l0 ♀, 0 ♂, 0 n, 0 l
If0 ♀, 0 ♂, 1 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l
400EBWIr5 ♀, 9 ♂, 18 n, 0 l0 ♀, 0 ♂, 0 n, 0 l4 ♀, 2 ♂, 52 n, 14 l1 ♀, 1 ♂, 6 n, 0 l15 ♀, 15 ♂, 15 n, 0 l0 ♀, 2 ♂, 3 n, 0 l
600EBWIr3 ♀, 12 ♂, 26 n, 0 l0 ♀, 0 ♂, 2 n, 0 l0 ♀, 0 ♂, 48 n, 24 l0 ♀, 0 ♂, 1 n, 0 l0 ♀, 1 ♂, 26 n, 1 l1 ♀, 1 ♂, 4 n, 0 l
800EBWIr4 ♀, 6 ♂, 98 n, 0 l0 ♀, 0 ♂, 9 n, 20 l1 ♀, 1 ♂, 22 n, 195 l1 ♀, 1 ♂, 47 n, 0 l1 ♀, 7 ♂, 42 n, 6 l1 ♀, 3 ♂, 19 n, 4 l
1000PBFIr3 ♀, 2 ♂, 36 n, 1 l0 ♀, 0 ♂, 0 n, 3 l2 ♀, 2 ♂, 117 n, 5 l0 ♀, 0 ♂, 5 n, 0 l10 ♀, 12 ♂, 39 n, 0 l0 ♀, 0 ♂, 11 n, 0 l
PAP200SOFDIr0 ♀, 0 ♂, 18 n, 57 l0 ♀, 1 ♂, 5 n, 0 l1 ♀, 0 ♂, 62 n, 21 l0 ♀, 0 ♂, 0 n, 0 l2 ♀, 1 ♂, 1 n, 0 l0 ♀, 1 ♂, 3 n, 0 l
Hc0 ♀, 0 ♂, 2 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 1 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l
400EBWIr1 ♀, 1 ♂, 5 n, 10 l0 ♀, 1 ♂, 1 n, 0 l0 ♀, 0 ♂, 23 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l
600EBWIr0 ♀, 0 ♂, 3 n, 0 l0 ♀, 0 ♂, 3 n, 0 l0 ♀, 0 ♂, 13 n, 63 l0 ♀, 0 ♂, 3 n, 6 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 1 n, 0 l
Hc0 ♀, 0 ♂, 0 n, 1 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 0 n, 0 l
800MEBBLAIr0 ♀, 0 ♂, 2 n, 6 l0 ♀, 0 ♂, 0 n, 0 l0 ♀, 0 ♂, 27 n, 20 l0 ♀, 0 ♂, 4 n, 3 l0 ♀, 0 ♂, 3 n, 0 l0 ♀, 1 ♂, 5 n, 1 l
Legend: GOK: Gorski Kotar; MED: Medvednica; PAP: Papuk; Sp.: species; S: spring; A: autumn; Ir: Ixodes ricinus; If: Ixodes frontalis; Hc: Haemaphysalis concinna; ♀ (female), ♂ (male), n (nymph), l (larva); EBDF: European beech forest with Deer fern (Blechno- fagetum sylvaticae Ht. 1950); EFDF: European fir forest with Deer fern (Blechno—Abietetum Ht. 1950); DBF: Dinaric beech–fir forest (Omphalodo—Fagetum Marinčak et al. 1992); MPF: Mixed private forest stand with European beech (Fagus sylvatica L.), European silver fir (Abies alba Mill.), Sycamore (Acer pseudoplatanus L.), European ash (Fraxinus excelsior L.) and Common hazel (Coryilus avellana L.); POEH: Pedunculate oak forest with European hornbeam (Carpino betuli—Quercetum roboris typicum Rauš 1969); EBW: European beech forest with Woodruff (Galio odorati-Fagetum Sougnez et Thill 1959); PBF: Pannonian beech–fir forest (Festuco drymeiae—Abietetum Vukelić et Baričević 2007); SOFD: Sessile oak forest with Festuca drymeia (Festuco drymeiae—Quercetum petraeae/Jank. 1968/Hruška 1974); MEBBLA: Mountain European beech forest with balm-leaved archangel (Lamio orvalae—Fagetum sylvaticae Ht. 1938).
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Vucelja, M.; Krčmar, S.; Habuš, J.; Perko, V.M.; Boljfetić, M.; Bjedov, L.; Margaletić, J. Altitudinal Distribution, Seasonal Dynamics and Borrelia burgdorferi Sensu Lato Infections in Hard Ticks (Acari: Ixodidae) in Different Forest Communities in Inland Croatia. Sustainability 2023, 15, 4862. https://doi.org/10.3390/su15064862

AMA Style

Vucelja M, Krčmar S, Habuš J, Perko VM, Boljfetić M, Bjedov L, Margaletić J. Altitudinal Distribution, Seasonal Dynamics and Borrelia burgdorferi Sensu Lato Infections in Hard Ticks (Acari: Ixodidae) in Different Forest Communities in Inland Croatia. Sustainability. 2023; 15(6):4862. https://doi.org/10.3390/su15064862

Chicago/Turabian Style

Vucelja, Marko, Stjepan Krčmar, Josipa Habuš, Vesna Mojčec Perko, Marko Boljfetić, Linda Bjedov, and Josip Margaletić. 2023. "Altitudinal Distribution, Seasonal Dynamics and Borrelia burgdorferi Sensu Lato Infections in Hard Ticks (Acari: Ixodidae) in Different Forest Communities in Inland Croatia" Sustainability 15, no. 6: 4862. https://doi.org/10.3390/su15064862

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