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Prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks (Acari, Ixodidae) in different Polish woodlands.

In 1996-1998, a total of 2285 Ixodes ricinus ticks (1063 nymphs, 637 males, 585 females) were collected from vegetation from 25 different localities in the 8 Polish provinces throughout the country. Ticks inhabited all 25 collection sites. The average number of ticks per collection site was 91.4 +/- 13.7. All 2285 ticks were examined for Borrelia burgdorferi sensu lato (s.l.) presence, of which 1333 specimens from 3 provinces were tested by routine indirect immunofluorescence assay (IFA) using polyclonal antibody PAB 1B29. The remaining 952 specimens from 5 provinces were examined by polymerase chain reaction (PCR), using FL6 and FL7 primers. The overall infection rate in ticks estimated by these 2 methods was 10. 2%. Nymphs showed lower positivity rate (6.2%) as compared to adult ticks (14.9% in females and 12.4% in males). The highest percentage of infected I. ricinus ticks (37.5%) was noted in the Katowice province while the lowest (4.1%) in the Bia ystok province. In particular collection sites, infection rates varied from 0-37.5%. The obtained results confirmed that B. burgdorferi s.l. is present throughout the distributional areas of I. ricinus in Poland and that a prevalence of spirochete-infected ticks may be high in some locations.

Animals↗

The occurrence DNA of Babesia microti in ticks Ixodes ricinus in the forest areas of Szczecin.

Babesia microti, an intraerythrocytic protozoan and the etiological agent of human babesiosis, is transmitted by the bite of the tick, Ixodes ricinus. The aim of the present study was to confirm the presence of B. microti by detection of the DNA of these protozoans. The prevalence of B. microti was studied using the PCR method with primers complementary to the gene fragment encoding nuclear small-subunit ribosomal RNA (ss-rDNA). In the course of this study a total of 2095 ticks, Ixodes ricinus, were examined. The mean infection rate was 6.2%. Variable prevalance values were also obtained from six different locations and they were further modified by the seasons of the year. The results confirmed the competence of I. ricinus as a vector of B. microti and that a B. microti-specific PCR can provide a sensitive test for laboratory detection of babesiosis.

Animals↗

Comparison of self-assessment and scutal index for the duration of Ixodes ricinus tick attachment.

Experimental studies have shown that the efficacy of transmission of B. burgdorferi sensu lato from an infected tick to an experimental animal is significantly associated with the duration of tick attachment. Scutal index is the ratio between body length and scutum width and was found to be an objective indicator of the duration of Ixodes scapularis tick attachment, but no data for I. ricinus have been published. In this preliminary report on 30 volunteers with an attached I. ricinus tick (21 had an adult female tick and 9 a nymph), none developed any sign of Lyme borreliosis but we were able to demonstrate two asymptomatic seroconversions in a six-week follow-up period. Twenty-six (87%) volunteers claimed that they were able to determine the approximate duration of tick attachment; participants with attached adult female ticks estimated the duration of attachment to be 30 (2-90) hours, while those with attached nymphs reported 48 (18-90) hours (p = 0.681). Scutal indices were 3.2 (2-5) for nymphs and 3.2 (1-5) for adult ticks, indicating 60- and 48-hour attachment, respectively. According to the volunteers' assessment and scutal index findings, approximately 80% and 45% of adult female ticks were removed within 48 hours after the tick bite (p = 0.032), respectively, while the corresponding values for nymphs were 85% and 17% (p = 0.053). Our findings were limited by low numbers of volunteers and by the lack of experimental data on the value of the scutal index for estimation of the duration of I. ricinus tick attachment.

Adolescent↗

The occurrence of Borrelia burgdorferi sensu lato in the populations of Ixodes ricinus in forest areas of Szczecin during 2000-2001.

The aim of the present study was to determine the seasonal abundance of ticks, Ixodes ricinus and their infection frequency with spirochetes, Borrelia burgdorferi in wooded areas of the vicinity of the city of Szczecin during 2000-2001. The ticks were collected from April to October. Presence of Borrelia burgdorferi sensu lato was detected with the method of indirect immunofluorescence (IFA). The results showed differences in seasonal activity of ticks within the years of the study. In the year 2000 a single peak of their increased activity was noted, while in the year 2001 no significant changes in the population of Ixodes ricinus were recorded. Borrelia burgdorferi s. l. was detected in 2000 and 2001 in 11.6 % and 9.6 % of the specimens captured, respectively, which constituted a statistically significant difference. The highest level of infection of Ixodes ricinus in 2000 was observed in April (13.2 %), while in 2001 - in July (12.7 %). The differences in the prevalence value between respective months of both years were not statistically significant. The results of the present study indicate a high risk factor of acquiring Lyme borreliosis by humans and animals in wooded areas of the Szczecin vicinity.

Animals↗

Detection of Borrelia burgdorferi sensu lato DNA in Ixodes ricinus ticks in North-western Poland.

In order to estimate the risk of contracting Lyme disease in the forest areas of north-western Poland, PCR-based studies were carried out on 6,817 Ixodes ricinus ticks for infection by the spirochaete B. burgdorferi sensu lato (s.l.). The studies were performed using the primers for the fla gene, conserved for all European genospecies of B. burgdorferi s.l. Based on the incidence of B. burgdorferi s.l. DNA in I. ricinus ticks at eight sampling sites during 1998-2001, it may be concluded that a risk of contracting Lyme disease is present in the forest areas of north-western Poland. The highest risk of infection (9.4 % of infected ticks) is posed by human contact with female I. ricinus, and the risk is higher in late spring and early summer than in late summer and early autumn. The north-western part of Poland is an endemic region for B. burgdorferi s.l.

Animals↗

[Ixodes ricinus, transmitted diseases and reservoirs].

The tick Ixodes ricinus has been recorded in most Italian regions especially in thermo-mesophilous woods and shrubby habitats where the relative humidity allow the tick to complete its 3 year developmental cycle, as predicted for the European climatic ranges. This tick acts both as vector and reservoir for a series of wildlife zoonotic pathogens, especially the agents of Lyme diseases, Tick borne encephalitis and Human Granulocytic Ehrlichiosis, which are emerging in most of Europe. To assess the spatial distribution of these pathogens and the infection risk for humans and animals within the territory of the Province of Trento, we carried out a long term study using a combination of eco-epidemiological surveys and mathematical modelling. An extensive tick collection with a GIS based habitat suitability analysis allowed us to identify the areas where tick occurs at various density. To identify the areas with higher infection risk, we estimated the values of R0 for Borrelia burgdorferi s.l., TBE virus and Anaplasma phagocytophila under different ecological conditions. We assessed the infection prevalence in the vector and in the wildlife reservoir species that play a central role in the persistence of these infections, ie the small mammals A. flavicollis and C. glareolus. We also considered the double effect of roe deer (Capreolus capreolus) which act as reservoir for A. phagocytophila but is an incompetent host for B. burgdorferi and TBE virus, thus reducing the infection prevalence in ticks of these last two pathogens. Infection prevalence with B. burgdorferi and A. phagocytophila in the vector was assessed by PCR screening 1212 I. ricinus nymphs collected by dragging in six main study areas during 2002. The mean infection prevalence recorded was 1.32% for B. burgdorferi s.l. and 9.84% for A. phagocytophila. Infection prevalence in nymphs with TBE virus, as assessed in a previous study was 0.03%. Infection prevalence in rodents was assessed by screening (with ELISA and PCR) tissues and blood samples collected from 367 rodent individuals trapped extensively during 2002 within 6 main study areas. A. flavicollis (N=238) was found to be infected with all three pathogens investigated, with infection prevalence ranging from 3.3% for TBE virus to 11.7% for A. phagocytophila, and 16.6% with B. burgdorferi s.l. C. glareolus (N=108) showed an infection prevalence of 6.5% with A. phagocytophila and 12.7% with B. burgdorferi s.l., while no individuals were infected with TBE virus. We also screened 98 spleen samples collected from roe deer with PCR, resulting in a mean prevalence of infection with A. phagocytophila of 19.8%. Using a deterministic model we explored the condition for diseases persistence under different rodent and roe deer densities. R0 values resulted largely above 1 for B. burgdorferi s.l. in the vast majority of the areas classified as suitable for I. ricinus occurrence in Trentino, while the condition for TBE persistence appeared to be more restricted by a combination of climatic condition and host densities.

Anaplasma phagocytophilum↗

[Capreolus capreolus and Ixodes ricinus as a reservoir of Bartonella in north-western Poland].

Capreolus capreolus and Ixodes ricinus as a reservoir of Bartonella in north-western Poland. The purpose of the study was to assess the prevalence Bartonella in Capreolus capreolus from north-western Poland forest. Supplementary, ticks infesting roe deer were also screened in order to ascertain their role as vectors and reservoirs of Bartonella. The samples of blood from 98 animals from north-western Poland were PCR-screened. Bartonella DNA was detected by using primers complementary to the intergenic spacer (ITS) between the 16S and 23S rRNA genes, which is used for identification of over a dozen species of this genus. Products of three different sizes were detected: 230 bp and 290 bp may represent two strains of B. capreoli, and 190 bp may be identify as B. bovis. All three amplicons were detected in blood, the 290 bp fragment from B. capreoli was present only in ticks, Ixodes ricinus. Generally, Bartonella infection in C. capreolus amounted to 21.4% of individuals, but was much higher during the autumn-winter seasons (62%), than in spring (4.3%). The results show that C. capreolus may be a reservoir for at least two species, i.e. B. capreoli and B. bovis, and probably do not cause persistent infection in roe deer. The high percentage of infested individuals during spring (84%) and infection detected in I. ricinus (5.2%) show that ticks are reservoir and vector of Bartonella.

Animals↗

PCR detection of granulocytic Anaplasma and Babesia in Ixodes ricinus ticks and birds in west-central Poland.

The aim of the study was to establish the role of forest birds as reservoirs of Anaplasma phagocytophilum and Babesia spp. in Wielkopolski National Park. A total of 108 birds from 9 species were collected between May-September 2002. Blood samples were taken from 84 specimens and 442 individuals of the common tick, Ixodes ricinus, were collected from the birds. The 73 additional ticks were collected from vegetation. PCR amplification of a fragment of the epank 1 gene and 18S rRNA gene was used for detection of A. phagocytophilum and Babesia spp. DNA, respectively. Pathogen DNA was not detected in any of the blood samples or ticks collected from birds. On the other hand, 3 ticks collected from vegetation (4.1% of all examined specimens) were positive for A. phagocytophilum DNA. In spite of the high level of infestation of birds by I. ricinus, it is clear that they do not constitute a competent reservoir of A. phagocytophilum and Babesia in WNP. Additionally, I. ricinus is not a significant vector in this area.

Anaplasma↗

[Occurence of pathogenic genospecies of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks collected from north-western Poland].

In order to learn the heterogeneity of the DNA of B. burgdorferi s.l. and the prevalence of co-infections of B. burgdorferi s.l. genospecies in the populations of I. ricinus, collected in north-western Poland, the nested PCR method was applied, a fragment of the fla gene being used as a marker. Basing on the prevalence data of B. burgdorferi s.l. DNA in I. ricinus ticks in 8 sampling sites during 1998-2001, it may be stated that a risk of contracting Lyme disease exists in forested areas of north-western Poland, the highest in relation to B. burgdorferi s.s. (76.3% infected ticks), lower by B. garinii (2% infected ticks), and minimal threat being posed by B. afzelii (0.3%). I. ricinus ticks collected in north-western Poland pose a risk of contracting double infection by B. burgdorferi s.l. genospecies, i.e. B. burgdorferi s.s. with B. garinii, and B. burgdorferi s.s. with B. afzelii. The north-western part of Poland represents an endemic area for B. burgdorferi s.l.

Animals↗

[Prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus population in forests of north-western Poland].

The ticks Ixodes ricinus were collected during three growing seasons in forested areas of north-western Poland. The ticks were collected by flagging at 30 sites. The specimens collected (nymphs and adults) were subjected, after an appropiate treatment, to indirect immunofluorescence analysis (IFA). In addition, effects of temperature and atmospheric precipitation on the tick infection with the spirochaete Borrelia burgdorferi were investigated based on data supplied by the Institute of Meteorology. A total of 8519 individuals of I. ricinus were collected at the sampling sites within May-October of three consecutive years. The pool of individuals obtained consisted of 7356 nymphs, 559 females, and 604 males. The B. burgdorferri prevalence varied rather widely, from 10 to 19%. Compared to nymphs, adult individuals showed a higher infection rate. A relationship between infection rate and I. ricinus activity in different years and months was studied in detail. The boreliosis risk was found to increase with increasing tick activity. On the other hand, no correlation was found between the meteorological parameters (temperature, precipitation), tick abundance, and the B. burgdorferi infection rate.

Animals↗

[Occurrence of Borrelia burgdorferi in Ixodes ricinus in The Netherlands].

In a 1989 surveillance to study the infection rate of Ixodes ricinus ticks with Borrelia burgdorferi, a total of 1838 nymphal and adult Ixodes ricinus ticks were collected from 20 locations in The Netherlands. By means of a culture method using modified Barbour-Kelly-Stoenner medium nymphs were examined in pools of 4 and adults were examined individually. With 63 out of 75 isolates propagation and serological identification were possible. All of these 63 isolates were identified as Borrelia burgdorferi by means of an indirect immunofluorescence assay. In nymphs the minimal infection rate was 2.4%. The rate of infection for adults was shown to be 14.3%. These results are in agreement with the percentages found by other investigators in West-Europe. In all locations examined Ixodes ricinus were found to be infected with Borrelia burgdorferi, although the infection rate varied. The results indicate that the infection is widespread among ticks in The Netherlands and the potential risk of acquiring Lyme borreliosis after a tick bite is serious.

Animals↗

An electron microscopic study of Cytoecetes phagocytophila infection in Ixodes ricinus.

Tick-borne fever, caused by a rickettsia-like organism, Cytoecetes phagocytophila, is transmitted by the sheep tick Ixodes ricinus. An electron microscopic technique was developed to examine I ricinus for C phagocytophila infection. Infected and uninfected ticks were obtained from a laboratory maintained tick culture. All stages of I ricinus collected from one field site were examined; 44 per cent of nymphae and 32 per cent of adults were infected with C phagocytophila, but larvae were uninfected. This supports the previously held theory of transtadial, but not transovarial transmission of tick-borne fever.

Animals↗

Immunity of dogs against Babesia canis, its vector tick Dermacentor reticulatus, and Ixodes ricinus in endemic area.

Previous epidemiological studies allowed us to accurately define endemic areas of canine babesiosis and tick distribution in southeastern France (Martinod, 1983). Using a micro-ELISA test 100 dogs sera were tested with 3 antigens: Babesia canis, Dermacentor reticulatus and Ixodes ricinus. Antibodies against B. canis and its vector D. reticulatus were detected in an endemic area, sometimes with high levels (optical density 1.38 and 0.80 respectively). A correlation factor and regression lines were found between ELISA activity of B. canis and vector tick antigens, even for dogs which never showed any babesiosis symptoms. These results were compared with those of an area without any babesiosis. Furthermore I. ricinus antigens detected ELISA activity in sera of dogs; some cross reactions were observed between I. ricinus and D. reticulatus antigen.

Animals↗

[Distribution of Ixodes ricinus ticks in Yugoslavia].

In this paper the authors present distribution of tick Ixodes ricinus in both republics of Yugoslavia--Serbia and Montenegro. The ticks were collected from domestic animals and humans sporadically. The determination showed following species: Ixodes ricinus, I. persulcatus, I. hexagonus, Dermacentor marginatus, D. pictus, Rhipicephalus bursa, R. sanguineus, Haemaphysalis punctata, H. inermis, H. sulcata, H. leporis-palustriis, H. concinna, Boophilus (margaporus) calcaratus, Hyalomma savignyi, H. excavatum, H. detritum and H. rufipes. The species I. ricinus has been detacted in all parts of the country.

Animals↗

[Comparison of misoprostol and ricinus oil meal for cervical ripening and labor induction].

OBJECTIVE: To compare the safety and efficacy of intravaginal prostaglandin E1, misoprostol with ricinus oil meal for labor induction. METHODS: Sixty patients with an indication for induction of labor were randomly assigned to two groups (30 cases each) induction with misoprotol, 50 micrograms intra-vaginally every 3 hours until active labor, or with that ricinus oil meal was taken. RESULTS: The time from start of induction to vaginal delivery was significantly shorter in the misoprostol group (12.2 vs 18.1, P < 0.05), and fewer patients in the misoprostol group required oxytocin augmentation (10.0% vs 40.0%, P < 0.05). The mean change in the Bishop score was significantly higher in those receiving misoprostol (5.5 vs 3.1, P < 0.05). Uterine tachysystole occurred more frequently in patients in the misoprostol group (16.7%) than in the ricinus oil meal group (3.0%). No significant differences were noted in the mode of delivery and patients of successful labor induction. CONCLUSIONS: Vaginally administered misoprostol is an effective agent for the cervical ripening and induction of labor.

Abortifacient Agents, Nonsteroidal↗

Amino acid sequence of a new 2S albumin from Ricinus communis which is part of a 29-kDa precursor protein.

The isolation and sequence determination of a new 2S albumin storage protein from Ricinus communis seeds denoted 2S ASP-Ib are described. The fragment approach using selective enzymatic cleavage, Edman degradation, and mass spectrometry was used to demonstrate that the 11-kDa heterodimer protein linked by disulfide bridges has the following structure: short chain, GEREGSSSQQCRQEVQRKDLSSCERYLRQSSS; long chain, <QQQESQQLQQCCNQVKQVRDECQCEAIKYIAEDQIQQGQLHGEESERVAQRAGEIVSSCGVRCMR . The molecular weight of the intact protein, 11,140 +/- 2, determined by matrix-assisted laser desorption mass spectrometry was consistent with the assigned structure. The S- and L-chains are identical to residues 18-49 and 66-130 of the precursor protein predicted by S. D. Irwin, J. N. Keen, J. B. C. Findlay, and J. M. Lord [(1990) Mol. Gen. Genet. 222, 400-408], on the basis of the structure of a cDNA isolated using probes based on the sequence of another 2S albumin, described by F. S. Sharief and S. S. L. Li [(1982) J. Biol. Chem. 257, 14753-14759], which we denote 2S ASP-Ia. Three of the four termini could have been produced by posttranslational processing by endopeptidase(s) and carboxypeptidase(s) which utilized basic residues as the cleavage sites. Mass spectrometric evidence suggested that the protein presented microheterogeneity at its termini, i.e., truncated forms presumably due to processing heterogeneity. The present characterization of the 2S ASP-Ib protein, the second 2S albumin from Ricinus communis seeds, demonstrates that the 237-residue precursor protein codes for two different heterodimer proteins containing 97 and 99 residues each. This system should be useful for studying the posttranslational processing of plant storage proteins.

Amino Acid Sequence↗

The Ricinus communis 2S albumin precursor: a single preproprotein may be processed into two different heterodimeric storage proteins.

The Ricinus communis (castor bean) 2S albumin is a heterodimer of glutamine-rich, disulphide-linked 4 and 7 kDa polypeptide. A cDNA library was constructed using mRNA from maturing castor bean endosperm as template. Clones containing sequences complementary to albumin mRNA were isolated by hybridization using as a probe a mixture of synthetic oligonucleotides representing sequences predicted for a peptide present in the 2S albumin large subunit. The nucleotide sequence contained an open reading frame encoding a preproprotein of 258 amino acid residues. The preproprotein included both polypeptides of the previously sequenced 2S albumin. In addition, this precursor included two further glutamine-rich sequences which, in term of their size and conserved cystein residues typically found in seed proteins of the 2S albumin superfamily, possible represent the small and large polypeptide subunits of a second heterodimeric storage protein. A post-translational processing scheme is proposed which would result in a single preproprotein generating two distinct heterodimeric 2S albumins. The generation of a second heterodimer seems likely since polypeptide candidates for its small and large subunits were found in the Ricinus 2S albumin fraction, and N-terminal protein sequencing confirmed the existence of the putative small subunit.

2S Albumins, Plant↗

Discovery of an extended bundle sheath in Ricinus communis L. and its role as a temporal storage compartment for the iron chelator nicotianamine.

The extended bundle sheath (EBS) is a specialized layer of cells that enhances the lateral transport of photoassimilates within the leaf. This little-known tissue is often considered to be legume-specific. We identified an EBS in cotyledons and leaves of the non-legume Ricinus communis L. By means of cytological and immunological studies and using the localization of the iron-chelator nicotianamine as an established indicator for mass transport, we confirmed its role as a transport tissue and a temporal sink. Observations on cotyledons of Ricinus seedlings further proved that the EBS carries out these tasks from a very early stage of development onwards. This is the first time that information has been obtained on the physiological role of an EBS in a non-legume. Our results support the idea of its widespread occurrence among higher plants.

Azetidinecarboxylic Acid↗