Competence of urban rats as reservoir hosts for Lyme disease spirochetes

To determine whether urban rats serve efficiently as reservoir hosts for the agent of Lyme disease, we recorded the frequency of infection in nymphal Ixodes ricinus (L.) ticks that had fed as larvae on experimentally infected Norway rats, Rattus norvegicus (Berkenhout), or on black rats, R. rattus (L.), and evaluated the nidicolous venue of transmission. Subadult vector ticks attached readily to Norway rats as well as black rats and virtually all became infected in the course of feeding. Larval ticks detached when these nocturnally active hosts were at rest. Rats appeared to be competent reservoir hosts of Lyme disease spirochetes in a transmission cycle in urban sites.     

Tick infestation patterns and prevalence of Borrelia burgdorferi in ticks collected at a veterinary clinic in Germany

A total of 4803 domestic and wild animals which were presented for examination at a veterinary clinic in north Baden, Germany over a period of 1 year were examined for tick infestation. A total of 434 nymphal and adult ticks were collected from 175 hosts. Ticks found belonged to the species Ixodes ricinus (385), Ixodes hexagonus (48), and Ixodes ventalloi (one). The polymerase chain reaction was used to examine 132 I. ricinus and 21 I. hexagonus for the presence of Borrelia burgdorferi. Twenty-two per cent of adult I. ricinus were infected as were one female and one larval I. hexagonus.     

Individual pharmacodynamics assessed by antilymphocyte action predicts clinical cyclosporine efficacy in psoriasis

The vector competence of 2 tick species, Ixodes ricinus (L.) and Ixodes scapularis Say, was determined and compared for 3 genospecies of Borrelia burgdorferi. The 3 genospecies of B. burgdorferi used in the following experiments were Borrelia burgdorferi sensu stricto (B-31 and B-31.D1 clone), Borrelia afzelii (strain Pgau. C3), and Borrelia garinii (strain VS286 and VSBP). Spirochetes from all 5 strains were inoculated intradermally into outbred mice; larval ticks of both species were subsequently fed on those mice and replete larvae were assayed for infection by culture in BSK-H media every 7 d for 4 wk. Infection frequencies in I. scapularis exposed to the 5 strains were as follows: B-31 (90%), B-31.D1 (83%), Pgau.C3 (87%), VS286 (10%), and VSBP (5%). The comparable infection frequencies for I. ricinus were B-31 (3%), B-31.D1 (3%), Pgau.C3 (90%), VS286 (5%), and VSBP (3%). Resultant nymphal I. scapularis successfully transmitted B-31, B-31,D1, Pgau.C3, and VS286 to outbred mice. I. ricinus nymphs transmitted Pgau.C3 and VS286. Both species failed to transmit strain VSBP.     

Involvement of serotonergic receptor subtypes in the production of antinociception by psychological stress in mice

The infestation of birds by immature Ixodes ricinus was studied during 6 months in a Swiss woodland, where Lyme borreliosis is endemic. Thirteen passerine species were found to be parasitized by I. ricinus subadults and specially Turdus merula, T. philomelos and Erithacus rubecula. Overall, 300 larvae and 162 nymphs were collected on 95 avian hosts. Prevalence of infestation of nymphs on birds was higher in spring; larvae peaked in summer. The infection of birds by Borrelia burgdorferi was also studied using blood cultivation and examination of ticks. Motionless spirochetes were isolated from two E. rubecula. Infected ticks were removed from five species of passerines, and mainly three species of Turdidae (T. merula, T. philomelos and E. rubecula). Infection rate of larvae and nymphs by spirochetes averaged 16.3% and 21.7%, respectively. These percentages, compared to the infection rate of questing ticks collected through dragging, suggest that some Turdidae may play a role as amplifying hosts for spirochetes in the focus.     

Infusion methods for determination of peripheral resistance: influence of infused medium and back pressure

Many isolates of Borrelia burgdorferi have been obtained from ticks and vertebrate tissues collected in North America and continental Europe but only one established culture of United Kingdom Borrelia burgdorferi has been recorded. In this paper we report the isolation of B. burgdorferi from one of 108 tick pools representing 733 ticks and eight-four tissue samples from twenty-six rodents collected in the U.K, and the subsequent failure to establish the isolate (from ticks collected in Fordingbridge) in culture. In contrast, using identical techniques and culture medium, B. burgdorferi was isolated from one of seven tick pools collected in Switzerland, and from a single pool of ticks collected in Slovakia, and both isolates were successfully passaged. Analysis of questing I. ricinus collected from Fordingbridge by direct immunofluorescence showed 6/32 (19%) of adults and 8/108 (7%) of nymphs were positive for B. burgdorferi, although only one nymph contained > or = 1000 spirochaetes. To examine further the problem of isolating U.K. B. burgdorferi, twelve Ixodes ricinus tick samples from Fordingbridge, a recognized focus of Lyme disease, were subjected to isolation and culturing techniques, and the procedures monitored by use of the polymerase chain reaction (PCR). Whereas 11/12 samples were PCR positive after 2 weeks in culture, only one was PCR positive after 4 weeks. Motile spirochaetes were not visible by dark-field microscopy in any of the cultures. The results indicate that the standard BSK II medium routinely used to isolate and culture B. burgdorferi does not readily support the replication of the Borrelia species endemic to the U.K.     

Seasonal variation in the capacity of the bank vole to infect larval ticks (Acari: Ixodidae) with the Lyme disease spirochete, Borrelia burgdorferi

Seasonal variation in the capacity of bank voles, Clethrionomys glareolus (Schreber), to infect larval Ixodes ricinus (L.) ticks with Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner was examined from May through October 1991 at Bogesund, near Stockholm in south-central Sweden. Although larval infestations of bank voles were greatest in June and July, nearly 70% of all larval ticks infected with spirochetes by bank voles at this site became infected during August and September. Seasonality of infectivity was related to the degree of earlier nymphal infestation on voles as well as to the age composition of the vole population. These factors may influence the infectivity of other rodent reservoirs of B. burgdorferi, both in Europe and North America. Moreover, in determining the reservoir potential of tick hosts, a host population's spirochetal infectivity should be determined for the entire period of larval infestation rather than just during the period of peak larval infestation.     

Temporal correlations between tick abundance and prevalence of ticks infected with Borrelia burgdorferi and increasing incidence of Lyme disease

The abundance of host-seeking Ixodes scapularis nymphs, the principal vector for the Lyme disease agent, Borrelia burgdorferi, in Old Lyme, Lyme, and East Haddam, Connecticut, was compared with the incidence of reported human Lyme disease in the 12-town area around the Connecticut River and the State of Connecticut for the period 1989 to 1996. Ticks were sampled from lawns and woodlands by dragging flannel over the vegetation and examined for the presence of B. burgdorferi by indirect fluorescent antibody staining. The infection rate of the nymphal ticks by B. burgdorferi during the 9-year period was 14.3% (of 3,866), ranging from 8.6% (1993) to 24.4% (1996). The incidence of Lyme disease was positively correlated with tick abundance in the 12 town area (r = 0.828) and the State of Connecticut (r = 0.741). An entomological risk index based upon the number of I. scapularis ticks infected by B. burgdorferi was highest in 1992, 1994, and 1996 and was highly correlated with the incidence of Lyme disease in Connecticut (r = 0.944). The number of Lyme disease cases has been influenced, in part, by annual changes in population densities of I. scapularis and, presumably, a corresponding change in the risk of contact with infected ticks. Based upon tick activity and spirochetal infection rates, epidemiologically based Lyme disease case reports on a regional scale appear to reflect real trends in disease.     

Measurement of apoptotic fragments in growing hair follicles following gamma-ray irradiation in mice

Feeding ticks are generally spatially distributed in clusters on vertebrate hosts. To test the effect of clustering on transmission of a tick-borne pathogen, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner-infected Ixodes ricinus L. nymphs and uninfected I. ricinus larvae were allowed to feed together in retaining chambers on uninfected AKR/N mice. Engorged infective nymphs dropped off at days 5, 6, and 7, and the 1st infected larvae that fed in the chambers together with the infected nymphs dropped off at day 5. In contrast, ear biopsies and xenodiagnostic larvae placed on the head remained negative during that period. These results suggest that a cofeeding transmission occurred between B. burgdorferi-infected ticks and noninfected ones in the absence of a disseminated infection. Further investigations are being undertaken to determine whether the mechanism responsible for this cofeeding transmission is similar to that described previously with virus-infected ticks.     

Mycobacterium tuberculosis reactive T cell clones from naturally converted PPD-positive healthy subjects: recognition of the M. tuberculosis 16-kDa antigen

The methods of spatial statistics were applied to assess the geographical pattern of risk of Lyme borreliosis in Central Bohemia, the Czech Republic, based on retrospective data on disease contractions. The statistical risk was then compared at 15 selected localities with the infection challenge presented by ticks and insects carrying borreliae. Over 5,000 Ixodes ricinus (L.) ticks and 390 haematophagous dipterans were screened by direct immunofluorescence method, and the spatial and seasonal variance of infection rates were studied. Infected ticks were found at each locality throughout the warm season; in nymphs, sample infection rates ranged from 4.9% to 23.1% with a mean of 14.5% in spring, from 7.7% to 28.7% with a mean of 16.1% in summer, and from 7% to 20.6% with a mean of 13.6% in autumn. The statistical risk was found to correlate well with an average nymphal infection challenge, i.e. I. ricinus nymphal abundance x infection rate, at a given locality. Statistically significant cumulation of insect-history recalling patients into several, generally wetland, areas was ascertained; borreliae were revealed in 0.5% of the dipterans examined.     

Infection rate of Ixodes ricinus ticks with Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi sensu stricto in Slovenia

In spring 1993, Ixodes ricinus ticks were collected from six regions of Slovenia to determine their overall rate of infection with Borrelia burgdorferi sensu lato and to assess the frequency of individual species in these tick populations. Ticks were dissected and midgut tissue inoculated into modified Barbour-Stoenner-Kelly (BSK II) medium. Borrelia isolates were differentiated into separate species using species-specific polymerase chain reaction (PCR) primers and by large restriction fragment pattern (LRFP) analysis. Infected ticks were found in all six regions surveyed. Spirochaetes were isolated from 69 of 363 ticks (19%): the isolation rate from adult female ticks was 35% (23/66 ticks cultured), from adult male ticks 22% (20/91), and from nymphal ticks 13% (26/206). Determination of the species of 60 isolates revealed that 32 were Borrelia afzelii (53%), 20 were Borrelia garinii (33%), and 8 were Borrelia burgdorferi sensu stricto (13%). In the Ljubljana region Borrelia afzelii and Borrelia garinii predominated (43% and 40%, respectively), whereas Borrelia burgdorferi sensu stricto constituted only 17% of isolates. In three other regions of the country Borrelia afzelii was isolated exclusively, although the number of isolates investigated was small. This study demonstrates the presence of all three European species of Borrelia burgdorferi sensu lato within the Slovenian tick population and also within a geographic area of less than 100 m2.     

The timing of color and location processing in the motor context

The contribution of migratory and resident birds to the introduction of Lyme disease will vary with the degree to which various species expose themselves to, and are infested by, juvenile vector ticks, and their ability to support and transmit the infectious agent. To examine the relative contribution of various passerine species during the emergence of this disease, we compared the abundance and infection rates of the blacklegged tick, Ixodes scapularis Say, removed from mist-netted birds with those from live-trapped mice at a coastal study site in southern Maine, collected during an 8-yr period in which the range of this tick and the incidence of Lyme disease increased in the state. Weekly bird-banding sessions using six 12-m Japanese mist nets were carried out from May through August 1989-1996. In 1989, 1991 and 1993, mice were live-trapped in a Sherman trap grid (7 by 7 m) during five 3-night sessions, June through August; in 1994-1996, 2 such grids were similarly trapped. Annual adult tick abundance was estimated by flagging vegetation. We removed 2,633 juvenile deer ticks from 1,713 of 1,972 birds examined. Twenty-five of 64 bird species were infested. The percentages of birds infested and the rate of infection among removed larvae and nymphs increased over the years, but species varied markedly in their ability to infect ticks. No infected larvae were removed from catbirds or towhees. The larval to nymphal ratio was higher in mice than in birds. Infection rates among bird-derived larvae were less than among mice-derived larvae, but increased with time. Because of the different ways in which individual species of passerine birds contribute to the availability of vector ticks and respond to the agent of Lyme disease in emerging areas, further research into host competency and borreliacidal mechanisms is needed.     

A method and device for measuring force transfers between the deep flexors in the musician''s hand

The efficacy of passive immunisation against tick-transmitted Lyme disease spirochaetal infection was determined in relation to the duration of previous feeding of infected vector ticks. Thus, mice challenged with spirochaete-infected unfed or partially fed nymphal ticks were passively immunised with monoclonal and polyclonal antibodies against the Lyme disease spirochaete (Borrelia burgdorferi) at various intervals after tick attachment. Spirochaetal infection in challenged mice and engorged ticks was verified by xenodiagnosis and indirect immunofluorescent antibody assay, respectively. Although tick-transmitted spirochaetal infection could be aborted by anti-OspA antibodies and hyperimmune antiserum, nearly all immunised mice challenged with infected ticks that had previous 36-h attachment became infected. More than 72% of the nymphal ticks used in this challenge retained their B. burgdorferi infection after engorgement on mice immunised with anti-spirochaete antibodies, and their subsequent infectivity to mice remained effective. It is concluded that a higher efficiency of transmission by partially fed infected nymphs and a lower efficacy of passive immunisation against infection result from an effect of previous feeding of infected ticks that activates antigenic change and enables the spirochaetes to circumvent OspA-based humoral immunity.     

Geographic risk for lyme disease and human granulocytic ehrlichiosis in southern New York state

Ixodes scapularis, the tick vector of Lyme disease and human granulocytic ehrlichiosis (HGE), is prevalent in much of southern New York state. The distribution of this species has increased, as have reported cases of both Lyme disease and HGE. The unreliability of case reports, however, demonstrates the need for tick and pathogen surveillance in order to accurately define areas of high risk. In this study, a total of 89,550 m2 at 34 study sites was drag sampled in 1995 and a total of 51,540 m2 at 40 sites was sampled in 1996 to determine tick and pathogen distribution in southern New York state. I. scapularis was collected from 90% of the sites sampled, and regionally, a 2.5-fold increase in nymphal abundance occurred from 1995 to 1996. I. scapularis individuals from all sites were infected with Borrelia burgdorferi in 1995, while an examination of ticks for both B. burgdorferi and the agent of HGE in 1996 confirmed that these organisms were present in all counties; the average coinfection rate was 1.9%. No correlation was found between estimated risk and reported cases of Lyme disease. The geographic disparity of risk observed among sites in this study underscores the need for vector and pathogen surveillance on a regional level. An entomologic risk index can help identify sites for targeted tick control efforts.     

Effects of the feeding process of Ixodes scapularis (Acari: Ixodidae) on embryonic development of its parasitoid, Ixodiphagus hookeri (Hymenoptera: Encyrtidae)

The chalcid wasp Ixodiphagus hookeri (Howard) is a parasitoid of several ixodid ticks including the blacklegged tick, Ixodes scapularis Say. We evaluated effects of the feeding process of nymphal I. scapularis on the embryonic development of I. hookeri. Potentially wasp-parasitized nymphal I. scapularis were collected on Prudence Island, RI. Subsamples of the questing nymph cohort were allowed to feed on laboratory white mice. Both the body length and the scutal length of ticks were measured individually for questing nymphs and for feeding nymphs that were removed from hosts at time intervals of 12, 24, 36, 48, and 72 h after attachment. The diameters of wasp eggs they contained were also measured for each designated time interval. There was a positive relationship between the mean scutal index (ratio between body length and scutal length) of ticks and the mean diameter of wasp eggs during 72 h of tick feeding (P < 0.05). Moreover, it appeared that within 24 h of tick attachment, the scutal index of ticks remained unchanged. However, after that period, the scutal index increased significantly (P < 0.05). Diameters of wasp eggs increased continuously during tick feeding and at 72 h after attachment, enclosed eggs and completely formed larvae were found in several ticks. We conclude that factors related to the feeding process of nymphal I. scapularis are necessary to initiate the embryonic development of wasps.     

Comparison of polymerase chain reaction with culture and enzyme immunoassay for diagnosis of pertussis

The risk for human infection with Lyme disease appears linked to the abundance of infected vector ticks, principally Ixodes dammini Spielman, Clifford, Piesman & Corwin, in the eastern United States. Habitat destruction by burning, although not well studied, has long been considered as an effective alternative to synthetic insecticides as a means of reducing tick populations. We evaluated the effect of a single spring burning of the woodland understory on the transmission risk of Lyme disease spirochetes (Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner) on Shelter Island, Long Island, NY. Following a burn in early April 1991, the abundance of nymphal I. dammini was 49% lower in the burned portion of a woodlot compared with the unburned portion. However, risk of encountering nymphs infected with B. burgdorferi remained similar in both burned and unburned woods. It is suggested that burning vegetation may disproportionately kill deer-derived rather than rodent-derived nymphs, significantly reducing tick abundance without affecting transmission risk.     

Reduced abundance of Ixodes scapularis (Acari: Ixodidae) with exclusion of deer by electric fencing

To assess the effect of deer exclusion on populations of Ixodes scapularis Say (formerly I. dammini Spielman, Clifford, Piesman & Corwin) in the northeastern United States, host-seeking ticks and ticks on white-footed mice, Peromyscus leucopus (Rafinesque), were monitored inside and outside a wooded, residential deer exclosure (approximately 3.5 ha) in Lyme, CT, in 1991 and 1992. Another deer exclosure was added in Lyme (approximately 7.4 ha) during 1992. Additional sample sites at other residences served as secondary controls. A seven-wire, slanted, high-tensile electric deer fence was used at both areas. Larvae of I. scapularis were 81.5% (1991) and 97.8% (1992) less abundant within the exclosure than immediately outside the deer exclosures. Nymphs of I. scapularis were 47.4% (1991) and 55.8% (1992) less abundant within the deer fence. The effect on adult ticks was mixed. No difference in tick abundance was seen at the 3.5-ha site. However, larvae, nymphs, and adults were 100, 83.8, and 74.1% less abundant, respectively, in plots at the 7.4-ha exclosure > or = 70 m from the deer fence and isolated from woodlands outside the fence by lawns, driveways, and buildings. The recovery of larvae and nymphs of I. scapularis from mice captured within the deer exclosures indicates that infestations of nymphs and adults are probably, at least in part, a result of the movement of these rodents. Based upon the number of nymphs per 100 m2 infected with Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, the causal agent of Lyme disease, there were 73 and 82% fewer infected nymphs within the deer exclosures in 1992 in comparison with the number of infected ticks outside the fence and at the secondary control sites, respectively. The exclusion of deer in conjunction with other tick control strategies in large areas could substantially reduce populations of I. scapularis and the risk of acquiring Lyme disease.     

Characterization of Borrelia burgdorferi isolated from different organs of Ixodes ricinus ticks collected in nature

Borrelia burgdorferi was isolated from 22 out of 133 adult Ixodes ricinus ticks collected from vegetation at two sites in Switzerland. From 17 ticks, spirochetes could be isolated from more than one organ. When the different isolates obtained from one tick were compared by SDS-PAGE analysis, differences in the protein profiles were observed in 8 cases. The isolates were further compared by immunological methods using mono- and polyclonal antibodies. Differences were observed in the proteins of 31-35 kDa and 18-25 kDa. Genetic divergence among isolates was evaluated by use of a B. burgdorferi specific gene probe for ospA. Correlation could be observed between immunological differences in OspA defined by monoclonal antibody LA31 and genetic variation of ospA as judged by restriction fragment length polymorphism (RFLP). Our findings indicate that systemic infection in unfed I. ricinus adults, as reflected by isolation of B. burgdorferi from multiple organs of one tick, is more frequent (8/22, 36%) than previously described (5%). Moreover, the presence of different B. burgdorferi phenotypes/genotypes in one tick is described for the first time. The findings may have bearings (i) on the time of tick attachment required for spirochete transmission since borreliae are already present in the salivary glands of systemically infected ticks at the beginning of the blood meal and (ii) perhaps also on the diversity of B. burgdorferi phenotypes inoculated by these ticks.     

Characteristics of garden dormice that contribute to their capacity as reservoirs for lyme disease spirochetes

To describe the contribution of garden dormice to the epizootiology of Lyme disease, we compared their reservoir capacity for these pathogens to that of other sympatric hosts. Garden dormice are trapped most abundantly during early spring and again during midsummer, when their offspring forage. They are closely associated with moist forests. Garden dormice serve as hosts to nymphal ticks far more frequently than do other small mammals. Spirochetal infection is most prevalent in dormice, and many more larval ticks acquire infection in the course of feeding on these than on other rodents in the study site. Mature dormice appear to contribute more infections to the vector population than juveniles do. Replete larval ticks generally detach while their dormouse hosts remain within their nests. The population of garden dormice contributes five- to sevenfold more infections to the vector population than the mouse population does. Their competence, nymphal feeding density, and preference for a tick-permissive habitat combine to favor garden dormice over other putative reservoir hosts of Lyme disease spirochetes.     

Borrelia burgdorferi sensu lato in Russia and neighbouring countries: high incidence of mixed isolates

A total of 365 isolates of Borrelia burgdorferi sensu lato from 12 major administrative territories of Russia (from St. Petersburg in the west to South Sakhalin in the east) and from the Czech Republic, Estonia, Lithuania, Byelorussia, Moldavia, Ukraine and Kirghizia were identified by analysis of restriction polymorphism of ribosomal rrf-rrl spacer amplicons. The isolates were obtained mainly from ixodes persulcatus and I. ricinus ticks. Other sources included small mammals, human patients and I. trianguliceps ticks. The results showed that B. garinii (two variants) together with B. afzelii circulated throughout the territories studied. The distribution of the variant NT29 of the species B. garinii, the most frequently isolated, was associated with that of I. persulcatus ticks. B. burgdorferi sensu stricto, and the species B. valaisiana and B. lusitaniae (formerly the genomospecies VS116 and PotiB2, respectively) were isolated only from I. ricinus ticks in the western part of the studied territories. None of these three species were found in 327 isolates from Russia where I. persulcatus is the most frequently distributed vector. This work also provides evidence for a high incidence of mixed Borrelia infections within vectors and hosts (9.3% of isolates were mixtures of Borrelia species). A detailed analysis of Borrelia species distribution over the territories studied is presented.