Alicja Buczek

Borrelia burgdorferi s.l. in Ixodes ricinus (Acari: Ixodidae) Ticks Collected from Vegetation and Small Rodents in Recreational Areas of the City of Poznań

Abstract During 1998–1999, Ixodes ricinus (L.) populations were investigated in three different biotopes (deciduous, mixed, coniferous forest) situated in popular recreational areas in Poznań, Poland. In total, 1,123 questing ticks (1,002 nymphs, 69 males, 52 females) were collected by flagging vegetation. Additionally, in 1998 between May and September small rodents were trapped and inspected for feeding ticks. Altogether, 213 rodents of three species: Apodemus agrarius Pall., A. flavicollis Melchior, Clethrionomys glareolus Schreber were captured. Of 323 engorged ticks, 304 were larvae and 19 nymphs. All ticks collected from vegetation, as well as from rodents were examined for the presence of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigenwalt & Brenner s.l. spirochetes by indirect immunofluorescence assay (IFA) using PAB 1B29. The seasonal pattern of activity of questing I. ricinus was always bimodal (May/June and August/September). The most abundant tick population occurred in the deciduous forest. The total infection rate in questing ticks was 16.2%. Differences in mean infection prevalence of host-seeking ticks between three biotopes each year were not significant. On average more larvae parasitized on the genus of Apodemus than on C. glareolus. 17.8% of larvae and 31.6% of nymphs fed on rodents harbored spirochetes. The three rodent species contributed to a different degree in to transmission of the pathogen to subadult stages. ≈27% of larvae infested on A. agrarius, 22% on C. glareolus, and only 4.2% on A. flavicollis contained spirochetes. The results suggest that the prevalence of A. agrarius and C. glareolus in disturbed urban forests used for leisure activities seems to be crucial for the maintenance of B. burgdorferi s.l. in I. ricinus populations.

Babesia microti in Adult Dermacentor reticulatus Ticks from Eastern Poland

A total of 468 adult Dermacentor reticulatus ticks (298 females and 170 males) collected by flagging in the forests of Lubelskie province, eastern Poland, were analyzed by PCR and nested-PCR for the presence of Babesia microti DNA. In all, 21 ticks (4.5%) were found to be infected with B. microti. The infection rate in females (5.0%) was slightly greater than in males (3.5%). Detection of B. microti for the first time in adult D. reticulatus ticks suggests that this species should be considered as a potential vector of B. microti in Europe.

Questing behavior of Dermacentor reticulatus adults (Acari: Amblyommidae) during diurnal activity periods in eastern Poland.

ABSTRACT Studies on diurnal activity and factors affecting the questing behavior in Dermacentor reticulatus (F.) adults were conducted in an open-type habitat in eastern Poland (51° 15′ N, 22° 36′ E). Observations, for 25 min each, were made at 2-h intervals between 1000 and 1800 hours, during the autumn (from late September to mid-October 2000) and spring (from late April to mid-May 2001) activity peaks. The questing behavior of adult D. reticulatus ticks depended on the time of the day and season. The autumn activity was higher than that in spring, yet both these periods were characterized by a rising trend from the morning to early afternoon with a peak at ≈1400 hours (on average 40 and 30 specimens per one collection, respectively). In spring, the high activity (on average 31.8 specimens/ collection) persisted until late afternoon (1800 hours), while in the autumn period it declined before nightfall (on average 21.4 specimens/collection at 1800 hours). Females predominated over males in both seasonal periods of activity and greater differences in the sex ratio were recorded in the spring. We found a negative correlation between the diurnal activity of the adult ticks and temperature and between the number of active ticks and the length of day both in the autumn and spring. However, no statistically significant correlation was demonstrated between the diurnal activity of adult ticks and humidity. The investigations show that, in addition to environmental factors, tick host-seeking behavior is dependent on the activity of their hosts and biological traits of this species.

Origin of Alimentary Tract in Embryogenesis of Ixodes ricinus (Acari: Ixodidae)

Abstract We studied the embryos of Ixodes ricinus (L.) in the second and third trimester of embryonic development, by using light and transmission electron microscopy. At the beginning of the second trimester, the formation of the foregut and rectal sac, by a process of invagination, was observed. The invagination, which develops into the primordium of the hindgut, forms only in the third trimester. The rectum forms in the last phase of embryogenesis. The development of the midgut is incomplete during embryogenesis. The yolk is surrounded by a wall, formed of an amorphous basal lamina and flattened cells, that gradually accumulate deutoplasmic material. These cells do not acquire the typical features of the gut epithelium until after larval hatching. These features are, however, found in the cells forming the rectal sac.

Prevalence of infections and co-infections with 6 pathogens in Dermacentor reticulatus ticks collected in eastern Poland

Occurrence of co-infections with various pathogens in ixodid ticks creates a risk of increased severity of tick-borne diseases in humans and animals exposed to bite of the ticks carrying multiple pathogens. Accordingly, co-infections in ticks were subject of numerous analyses, but almost exclusively with regard to Ixodes ricinus complex whereas potential tick vectors belonging to other genera were much less studied. Taking into consideration the role of Dermacentor reticulatus in the transmission of various pathogens, we carried out for the first time the comprehensive statistical analysis of co-infections occurring in this tick species. An attempt was made to determine the significance of the associations between 6 different pathogens occurring in D. reticulatus (Tick-borne encephalitis virus = TBEV, Anaplasma phagocytophilum, Rickettsia raoultii, Borrelia burgdorferi s. l., Babesia spp., Toxoplasma gondii), using 2 statistical methods: determination of Odds Ratios (ORs) and the Fishers exact test. 634 questing Dermacentor reticulatus ticks (370 females and 264 males) were collected in 2011- 2013 by flagging the lower vegetation in 3 localities in the area of Łęczyńsko-Włodawskie Lakeland, situated in the Lublin region of eastern Poland. The presence of individual pathogens was detected by PCR. Ticks were infected most often with Rickettsia raoultii (43.8%), less with TBEV (8.5%), and much less with Babesia spp., Toxoplasma gondii, Borrelia burgdorferi s.l., and Anaplasma phagocytophilum (2.5%, 2.1%, 1.6% and 1.1%, respectively). The locality-dependent variability proved to be significant for TBEV (c2=11.063; P=0.004) and Toxoplasma gondii (c2=11.298; P=0.0035), but not for other pathogens. Two hundred seventy (42.6%) of the examined ticks were infected only with a single pathogen, and 54 (8.5%) showed the presence of dual co-infections, each with 2 pathogens. The most common were dual infections with participation of Rickettsia raoultii (7.41%); next, those with participation of the TBEV (5.21%), Toxoplasma gondii (1.58%), Borrelia burgdorferi s.l. (1.26%), Anaplasma phagocytophilum (0.95%), and Babesia spp. (0.63%). On the total number of 15 possible associations, in 9 cases co-infections occurred whereas in 6 cases they were not detected. The most noteworthy were positive co-infections with the participation of TBEV, which proved to be weakly significant (0.05<P<0.1) in associations with Toxoplasma gondii and Anaplasma phagocytophilum, with Odds Ratios over 3.3 and 4.4, respectively. The values of Odds Ratios exceeded 3.0 also at the co-infections of Rickettsia raoultii with B. burgdorferi s.l., and T. gondii with Babesia spp., but these associations did not attain a significance level. The co-infections of Rickettsia raoultii with Babesia spp. appeared not to be significant (0.05<P<0.1) with OR below 0.3. In conclusion, co-infections with various pathogens in D. reticulatus ticks seem to be relatively rare and mostly not significant.

Stray dogs and cats as potential sources of soil contamination with zoonotic parasites

INTRODUCTION AND OBJECTIVE The main source of many zoonoses is soil contaminated with feline and canine faeces. Thus, the aim of this study was to estimate the prevalence of intestinal parasites in stray dogs and cats adopted in Lodz shelter (Poland). MATERIALS AND METHODS In total, 163 faecal samples were collected from 95 dogs and 68 cats from 2011 to 2012. The samples were processed by sedimentation techniques using Mini Parasep®SF. RESULTS Six parasite genera belonging to protozoa, cestoda, and nematoda, were found in dogs, while eight were found in cats. Out of the 163 fecal samples, 37.4% were positive for the presence at least one species of intestinal parasites. The majority of positive dog samples contained eggs from Toxocara and Trichuris genera, and the family Ancylostomatidae, while Toxocara and Taenia eggs, as well as Cystoisospora oocysts, predominated in cat faeces. A significantly higher prevalence of parasites was noted in cats (48.5%) than in dogs (29.5%) (χ2=6.15, P=0.013). The Toxocara genus was the most prevalent parasite in both populations; eggs were found in 27.9% and 16.8% of cats and dogs, respectively. Animals younger than 12 months of age showed higher infection rates with Toxocara, but differences were not statistically significant. The average numbers of Toxocara eggs/gram of faeces in positive puppy and kitten samples were over 5 and 7 times higher than in older dogs and cats, respectively. Mixed infection were found in dogs (5.3%) and cats (8.8%). CONCLUSIONS Cat faeces represent a more important potential source of environmental contamination with zoonotic parasites than dog faeces. Among the detected parasites of stray dogs and cats, Toxocara present an important zoonotic risk for the local human population, especially children.

The toxic effect of permethrin and cypermethrin on engorged Ixodes ricinus females

INTRODUCTION Ixodes ricinus tick is of great medical and veterinary importance and has a wide range of geographical distribution. The study presents the effect of permethrin (Per) and cypermethrin (CM) on engorged I. ricinus females. MATERIALS AND METHOD The effect of perythroids studied on engorged I. ricinus females was assessed on the basis of the pre-oviposition and oviposition period. Remote effects of Per and CM application were assessed by investigation of the length and course of embryonic development and larval hatching from eggs laid by pyrethroid-treated females. Per (Copex WP) was used at doses of 0.78125-25.0 µg/1 specimen, and CM (Kordon 10WP) was applied at 0.3125-10.0 µg/1 specimen. Immediately after the feeding period, I. ricinus females were sprayed with 20 µl of a pyrethroid solution and kept at 28 °C and 75%RH. RESULTS The experiments demonstrated that CM exerted a stronger toxic effect on I. ricinus females than Per. The lowest doses of CM doubled the length of the pre-oviposition period while its highest doses prolonged the period nearly three times compared with the control. The pyrethroids applied reduced the number and weight of eggs and changed the parameters of the oviposition process. Application of the tested pyrethroid doses led to disturbances in the embryonic development of I. ricinus, i.e. the development was prolonged, few normal larvae hatched, numerous eggs and embryos at various developmental stages died, and larval hatch was inhibited. CONCLUSIONS Knowledge about the sensitivity of engorged females to different doses of the tested pyrethroids and the remote effects of their action can be used in practice for tick control among livestock animals, and the reduction of tick population abundance in the environment.

Factors influencing the distribution and activity of Dermacentor reticulatus (F.) ticks in an anthropopressure-unaffected area in central-eastern Poland

INTRODUCTION AND OBJECTIVES Despite the increasing prevalence and significant role of D. reticulatus in pathogen transmission, the factors influencing its distribution and activity are still poorly known. The paper presents for the first time a study of the D. reticulatus ecology in a protected area, in which the dependence of D. reticulatus occurrence and activity on the ecological habitat type is analysed. MATERIALS AND METHODS Tick collection and environmental monitoring were conducted from March - November 2012 and 2014 in five ecologically-diverse habitats in the Polesie National Park of central-eastern Poland. RESULTS The study shows that the most beneficial habitats for D. reticulatus are provided by meadow ecosystems dominated by plants from the class Molinio-Arrhenatheretea: Poa pratensis, P. trivialis, Festuca rubra, F. pratensis, and Alopecurcus pratensis, as well as Carex sp. sedges (64.78% of the tick collections). Less favourable conditions are offered by the fresh habitat covered by deciduous forest with dominance of Betula sp., A. pratensis, and Carex sp. sedges (15.27%), in the wet habitat dominated by Alnus glutinosa and Urtica dioica (9.00%), Betula humilis, Salix sp., and Carex sp. (8.44%), and in the subcontinental hornbeam forest Tilio-Carpinetum with poor undergrowth (2.50%). D. reticulatus avoids wet habitats with bog birch Betuletum pubescientis communities and moist pine forest Molinio-Pineteum, as well as habitats dominated by heather Calluna vulgaris. CONCLUSIONS The distribution and dynamics of the activity of adult D. reticulatus depends on the biological and geoclimatic conditions prevailing in tick habitats. The structure of the flora colonising a habitat may be an indicator of the presence of this tick species.

Head pediculosis in schoolchildren in the eastern region of the European Union.

OBJECTIVE The aim of the study was to investigate the prevalence of head pediculosis among children from state primary schools in the villages and towns in eastern Poland, one of the poorest regions of the European Union. MATERIALS AND METHODS The study was conducted in 2009-2012 in 30 rural and urban State schools. Current and previous results (1996-2000) of examinations performed in the study area were compared. The impact of socioeconomic factors on the prevalence of head pediculosis in eastern Poland in the period of 16 years was analysed. RESULTS Pediculosis was diagnosed in 2.01% of the schoolchildren, more frequently in pupils from rural (3.52%) than urban (0.98%) schools. Lice infestation was higher in girls (59.52%) than in boys (40.48%). The risk of pediculosis in children increased in schools that did not employ a nurse (mean 5.07%) and decreased when a part-time (mean 2.96%) or full-time nurse (mean 1.01%) was employed. The incidence of pediculosis doubled despite the improvement in the socioeconomic status of the population observed within the last 16 years. CONCLUSION The results indicate that personal hygiene behaviour may be of key importance in determining the spread of the disease.

Comparison of the toxic effect of pyrethroids on Ixodes ricinus and Dermacentor reticulatus females

INTRODUCTION Despite the increased rates of infestations with I. ricinus (Ir) and D. reticulatus (Dr) ticks observed over the last decade, no effective control methods have been developed so far. The present study was focused on assessment of the action of pyrethroids on these both tick species. MATERIALS AND METHOD The different doses of four pyrethroids, i.e. deltamethrin - D (K-Othrine), permethrin - P (Copex WP), cypermethrin - C (Kordon 10WP), and alphacypermethrin - AC (Alfasekt 5SC) were tested. The LD₅₀ for each tested compound was also determined for both tick species. Unengorged and engorged (maintained on rabbit skin) tick females were sprayed with 20 ml of 0.01563-0.50% solutions of the tested preparations. RESULTS The investigations showed that sensitivity of Ir and Dr to the tested pyrethroids, but the effects exerted by the different doses varied between both tick species and between engorged and unengorged females in these species. The strongest toxic effect on unengorged and engorged Ir and Dr females was exerted by D, whereas the effect of AC was weaker. The LD ₅₀ (in µg/1 g b.w.) of D, AC, C, and P for unengorged Ir and Dr females was, respectively, 55.4 and 25.5, 105.2 and 48.5, 225.9 and 197.7, and 553.8 and 380.8. In the case of engorged Ir and Dr females, the LD₅₀ of AC, D, C, and P reached a value of 0.9453 and 0.2310, 1.0428 and 1.3533, 3.489 and 6.5662, and 8.3955 and 7.3940, respectively. CONCLUSIONS The differences between the effects of the tested pyrethroids and their different doses on Ir and Dr highlight the necessity for development of a strategy for control of the tick species in different regions, based on investigations of their sensitivity to chemical compounds.