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.     

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.     

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.     

Simulation of management strategies for the blacklegged tick (Acari: Ixodidae) and the Lyme disease spirochete, Borrelia burgdorferi

A computer model (LYMESIM) was developed to simulate the effects of management technologies on populations of the blacklegged tick, Ixodes scapularis Say, and the Lyme disease spirochete Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner in eastern North America. Technologies considered in this study were area-wide acaricide, acaricide self-treatment of white-footed mice and white-tailed deer, vegetation reduction, and white-tailed deer density reduction. Computer simulations were run with normal weather patterns for coastal Connecticut and New York. Results showed that area-wide acaricide, vegetation reduction, or a combination of these technologies would be useful for short-term seasonal management of ticks and disease in small recreational or residential sites. Acaricide self-treatment of deer appears to be the most cost-effective technology for use in long-term management programs in large areas. Simulation results also suggested that deer density reduction should be considered as a management strategy component. Integrated management strategies are presented that could be used in pilot tests and operational tick and tick-borne disease programs.     

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.     

Accelerated infectivity of tick-transmitted Lyme disease spirochetes to vector ticks

We determined whether the span of infectivity of Lyme disease spirochetes (Borrelia burgdorferi) to vector ticks varies with the mode of infection in laboratory mice. Noninfected larval deer ticks were permitted to feed on two strains of spirochete-infected mice that had been naturally (via tick bite) and parenterally (via needle injection) infected with B. burgdorferi 2, 4, or 8 weeks earlier, and engorged ticks were dissected and examined for spirochetes by direct immunofluorescence microscopy. After initial infection, spirochetal infectivity to ticks was less efficient in needle-infected mice than in mice infected via tick bites. Tick-transmitted spirochetes develop more rapidly from the skin of infected mice and do not induce a strong antispirochete antibody response during the early stage of infection.     

Novel Borrelia burgdorferi isolates from Ixodes scapularis and Ixodes dentatus ticks feeding on humans

Seven cultures of Borrelia burgdorferi differing from strains B31 and ZS7 were identified from among 99 isolates from Ixodes scapularis ticks and from white-footed mice (Peromyscus leucopus) and 1 isolate from an Ixodes dentatus tick. Five of the six novel isolates from I. scapularis and the isolate from I. dentatus were from ticks feeding on humans. The six isolates from I. scapularis lacked OspA and OspB, four possessed an OspD band, and two reacted with an anti-OspC monoclonal antibody. Restriction fragment length polymorphisms of HindIII-digested DNAs from six OspA-negative isolates did not hybridize with radiolabeled ospA or LA88 DNA, and only isolate 46047 hybridized with the pG gene. Fragments similar to those recorded for the standard B. burgdorferi sensu stricto strains B31 and ZS7 were obtained with the fla and the HSP70 genes. Pulsed-field gel electrophoresis patterns of DNA digested with MluI included the specific B. burgdorferi sensu stricto band at 135 kbp for the five OspA-negative isolates from I. scapularis ticks. The six novel isolates apparently lack the 55-kbp plasmid encoding OspA. The pG-containing plasmid may be missing from all but isolate 46047. The isolate from the I. dentatus tick was similar to previous isolates from I. dentatus ticks feeding on rabbits. None of the isolates could be recovered from inoculated C3H/HeNCrlBR or white-footed mice. All isolates reacted with sera from humans with early or late Lyme disease. Our studies demonstrate that these borreliae occur in ticks feeding on humans, and therefore, at least some humans in the northeastern United States are likely being exposed to borreliae other than the classic B31-type strains that have thus far been isolated from humans.     

Scientific opportunities ... ''taken at the flood''

Rodent trapping and drag sampling in Van Cortlandt Park, New York City, yielded all stages of Ixodes scapularis, the deer tick vector of Lyme disease and human granulocytic ehrlichiosis (HGE). Polymerase chain reaction analyses of the ticks showed Borrelia burgdorferi and the Ehrlichia sp. that causes HGE.     

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.     

Transmission of Borrelia burgdorferi s.l. from mammal reservoirs to the primary vector of Lyme borreliosis, Ixodes ricinus (Acari: Ixodidae), in Sweden

Factors regulating prevalence of Borrelia burgdorferi s.l. Johnson, Schmid, Hyde, Steigerwalt & Brenner in Ixodes ricinus (L.) were examined during 1991-1992 at Bogesund, near Stockholm in south-central Sweden. Nine species of small and medium-sized mammals (Sorex araneus L., S. minutus L., Neomys fodiens Pennant, Clethrionomys glareolus [Schreber], Microtus agrestis [L.], Apodemus sylvaticus [L.], A. flavicollis [Melchior], Lepus europaeus Pallas, L. timidus L.) were found to infect feeding tick larvae with B. burgdorferi s.l., whereas two species of large mammals (Capreolus capreolus L., Alces alces L.) failed to infect feeding tick larvae with this spirochete. The most important mammalian reservoirs at the study locality were S. araneus and rodents, accounting for 91% of all I. ricinus larvae infected. In view of the great number of potentially effective reservoirs for B. burgdorferi s.l. in Sweden, control of Lyme disease by reduction of abundance of reservoir hosts will be difficult to achieve. We also found that infectivity of a rodent species is related to the number of infesting, potentially infective nymphal I. ricinus. Insectivores and rodents were the most important hosts of larval I. ricinus, whereas most nymphal ticks fed on hares and cervids. Adult I. ricinus were frequently found on all species of hares and cervids examined but never on insectivores and rodents. No single species seemed to be of paramount importance as a source of blood for female ticks. Therefore, control of Lyme disease by reduction of abundance of mammal hosts available for female tick engorgement will probably require massive reductions of numbers of both C. capreolus and L. timidus.     

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.     

Duration of Borrelia burgdorferi infectivity in white-footed mice for the tick vector Ixodes scapularis under laboratory and field conditions in Ontario

The duration of Borrelia burgdorferi infectivity in white-footed mice (Peromyscus leucopus) experimentally inoculated or infested with infected Ixodes scapularis nymphs was evaluated. Infectivity was assessed by infesting these mice with unfed I. scapularis larvae at 7, 21, 35 and 49 days post-inoculation (DPI) or post-infestation (PI). At 7 DPI, B. burgdorferi was transmitted from 18 of 24 syringe-inoculated mice and all three tick-infected mice to I. scapularis larvae which fed upon them. However, at 21, 35 and 49 DPI, significantly fewer mice were infective. Borrelia burgdorferi was isolated from tissues of 14 of 22 syringe-inoculated mice about 56 DPI, and from all three tick-infected mice. However, the level of agreement between xenodiagnosis and bacterial culture was no greater than would be expected by chance alone. We also determined if B. burgdorferi infectivity of mice varied in relation to periods of tick feeding in the field. White-footed mice were trapped during April, July and August 1993 from two habitats on Long Point peninsula (Ontario, Canada), where B. burgdorferi is endemic. Mice from each habitat were infested with laboratory-reared I. scapularis larvae. Ticks from each mouse were subsequently examined by immunofluorescent assay for B. burgdorferi infection and mice were cultured for B. burgdorferi. None of 3577 I. scapularis larvae fed on 62 mice captured within the cottonwood dune habitat were infected with B. burgdorferi, although it was isolated from six of these mice. Within the maple forest habitat, 0/24, 8/21 (38%) and 1/21 (5%) mice transmitted B. burgdorferi to I. scapularis larvae during April, July and August, respectively. Most mice from the maple forest with B. burgdorferi-positive tissues (14/21) were collected during July, although the level of agreement between xenodiagnosis and tissue culture was poor. Because B. burgdorferi infectivity in mice appears to be of short duration, overwintered I. scapularis larvae and nymphs may have to feed upon infected hosts at the same time of year in order for a cycle of B. burgdorferi infection to be maintained on Long Point. Infected I. scapularis nymphs, rather than persistently infected vertebrate hosts, likely serve as the overwintering "reservoir" for B. burgdorferi on Long Point.     

Geographic distribution of ticks (Acari: Ixodidae) in Michigan, with emphasis on Ixodes scapularis and Borrelia burgdorferi

A 12-yr (1985-1996) passive survey in Michigan based upon tick submissions from citizens yielded 4,755 ticks of 21 species, 16 of which were probably indigenous in the state. Three species of Dermacentor [most common, D. variabilis Say and D. albipictus (Packard)]; 2 species of Amblyomma [most common, A. americanum (L.)]; and 12 species of Ixodes (most common, I. cookei Packard and I. scapularis Say), as well as Haemaphysalis leporispalustris (Packard), Rhipicephalus sanguineus Latreille, and the soft ticks Ornithodoros kelleyi Cooley & Kohls, and Otobius megnini (Duges) were submitted. New state records were I. kingi Bishopp, I. texanus Banks, I. sculptus Neumann, and I. baergi Cooley & Kohls. Examination of gut smears from dissections of 1,037 ticks of 13 species by indirect immunofluorescent assay, using murine monoclonal H9724 as the primary antibody, revealed that 11 of 175 I. scapularis were infected with Borrelia spp. All positive I. scapularis were from Menominee County in the upper peninsula of the state, which also provided 79.8% of all submitted I. scapularis. Surveys for ticks on 5,449 hunter-killed white-tailed deer were conducted from 1988 to 1990, encompassed deer taken from 65 of the state's 83 counties, and showed that although D. albipictus was distributed widely in the northern part of the state, I. scapularis occurred only on deer taken from southern townships of Menominee County. Of 1,218 canine sera tested for antibodies to B. burgdorferi in 1992 and 1993, 25 of 299 (8.0%) from Menominee County were positive but only 1 of 919 sera submitted from 5 counties in the lower peninsula was positive.     

Temporal pattern of Borrelia burgdorferi p21 expression in ticks and the mammalian host

The temporal synthesis of the P21 protein of Borrelia burgdorferi and the development of the humoral response to this antigen was assessed in infected mice. p21 is a member of the ospE-F gene family and its protein, P21, has been shown to be expressed by B. burgdorferi within infected mice but not by spirochetes cultured in vitro. P21 was not detected on B. burgdorferi in unfed or engorged Ixodes dammini (also known as I. scapularis) ticks, further supporting the postulate that P21 synthesis is specific for the mammalian host. In B. burgdorferi-infected mice, ospE mRNA and OspE antibodies were observed at 7 d, whereas p21 mRNA and P21-specific antibodies were detected at 21-28 d, suggesting that p21 is expressed later than ospE. Moreover, ospA mRNA was not discernible until day 14, indicating that ospA, like p21, is not expressed in the early stages of tick-transmitted murine Lyme borreliosis. Because p21 is expressed during infection in mice, we assessed the human humoral response to P21. 28% (34 of 122) of the patients with either early- or late-stage Lyme disease, and 33% (11 of 33) of the individuals with Lyme arthritis had P21 antibodies, suggesting that a P21 response may serve, at least partially, as a marker of infection. Active immunization with recombinant P21 did not protect C3H mice from tick-borne B. burgdorferi infection, and passive transfer of P21 antiserum to infected mice did not alter the course of disease. These data suggest that the antigenic structure of B. burgdorferi changes during the early stages of murine infection.     

Reservoir competence of the meadow vole (Rodentia: Cricetidae) for the Lyme disease spirochete Borrelia burgdorferi

The reservoir competence of the meadow vole, Microtus pennsylvanicus Ord, for the Lyme disease spirochete Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner was established on Patience Island, RI. Meadow voles were collected from 5 locations throughout Rhode Island. At 4 of the field sites, M. pennsylvanicus represented only 4.0% (n = 141) of the animals captured. However, on Patience Island, M. pennsylvanicus was the sole small mammal collected (n = 48). Of the larval Ixodes scapularis Say obtained from the meadow voles on Patience Island, 62% (n = 78) was infected with B. burgdorferi. Meadow voles from all 5 locations were successfully infected with B. burgdorferi in the laboratory and were capable of passing the infection to xenodiagnostic I. scapularis larvae for 9 wk. We concluded that M. pennsylvanicus was physiologically capable of maintaining B. burgdorferi infection. However, in locations where Peromyscus leucopus (Rafinesque) is abundant, the role of M. pennsylvanicus as a primary reservoir for B. burgdorferi was reduced.     

Distribution and molecular analysis of Lyme disease spirochetes, Borrelia burgdorferi, isolated from ticks throughout California

Previous studies describing the occurrence and molecular characteristics of Lyme disease spirochetes, Borrelia burgdorferi, from California have been restricted primarily to isolates obtained from the north coastal region of this large and ecologically diverse state. Our objective was to look for and examine B. burdorferi organisms isolated from Ixodes pacificus ticks collected from numerous regions spanning most parts of California where this tick is found. Thirty-one isolates of B. burgdorferi were examined from individual or pooled I. pacificus ticks collected from 25 counties throughout the state. One isolate was obtained from ticks collected at Wawona Campground in Yosemite National Park, documenting the occurrence of the Lyme disease spirochete in an area of intensive human recreational use. One isolate from an Ixodes neotomae tick from an additional county was also examined. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis, agarose gel electrophoresis, Southern blot analysis, and the polymerase chain reaction were used to examine the molecular and genetic determinants of these uncloned, low-passage-number isolates. All of the isolates were identified as B. burgdorferi by their protein profiles and reactivities with monoclonal and polyclonal antibodies, and all the isolates were typed by the polymerase chain reaction as North American-type spirochetes (B. burgdorferi sensu stricto). Although products of the ospAB locus were identified in protein analyses in all of the isolates, several isolates contained deleted forms of this locus that would result in the expression of chimeric OspA-OspB proteins. The analysis of OspC demonstrated that this protein was widely conserved among the isolates but was also quite variable in its molecular mass and the amount of it that was expressed.     

Impact of controlled burns on the abundance of Ixodes scapularis (Acari: Ixodidae)

Information on the effect of vegetative destruction by controlled burns in reducing the abundance of the blacklegged tick, Ixodes scapularis Say, the vector for the agents of Lyme disease, human babesiosis, and human granulocytic ehrlichiosis, is limited. Therefore, the abundance of nymphal, larval, and adult I. scapularis was monitored by dragging the vegetation at 2 separate 4-ha tracts in Cockaponset State Forest in Connecticut following a single controlled burn on 15 April or 21 May 1992. The burn at the April burn site was rated as light to moderate with a flame height of 0.3 m and consumed approximately 67% of the surface leaf litter. The burn at the May burn site was rated moderate to severe with a flame height of 0.6-0.9 m., which consumed vegetation < 5 cm in diameter and approximately 100% of the surface leaf litter. The impact of the burn was strongly influenced by the intensity and timing of the burn. Burning of the vegetation resulted in a reduction of the abundance of nymphal I. scapularis by 74% at the moderately burned site and 97% at the severely burned site, compared with adjacent unburned woodland. No larvae were recovered later in the summer from the severely burned tract. However, judging by the comparable abundance of adult I. scapularis in the fall at the burned and unburned woodlands, the effect of the burns was temporary. Burning the vegetation for the control of I. scapularis appears limited in effect and could be applied only on a large scale in areas with little or no human habitations.     

Determining the duration of Ixodes scapularis (Acari: Ixodidae) attachment to tick-bite victims

The duration of tick attachment is one factor associated with risk for human infection caused by several tick-borne pathogens. We measured tick engorgement indices at known time intervals after tick attachment and used these indices to determine the length of time that ticks were attached to tick-bite victims in selected Rhode Island and Pennsylvania communities where the agents of Lyme disease and human babesiosis occur. The total body length and width as well as the length and width of the scutum were measured on nymphal and adult female Ixodes scapularis Say removed from laboratory animals at 0, 12, 24, 36, 48, 60, and 72 h after their attachment. Three engorgement indices were calculated at each time interval. In addition, engorgement indices measurements were recorded for 504 ticks submitted to a commercial laboratory for pathogen detection testing between 1990 and 1992. No detectable change was observed in the average engorgement indices for either nymphal or adult ticks between 0 and 24 h of attachment using any of the engorgement indices. After 24 h of tick attachment, all engorgement indices continuously increased: average indices for nymphs attached 36, 48, and 60 h were significantly different from those attached < or = 24 h and from each other. Similarly, average engorgement indices for adult ticks attached < or = 36 h were significantly different from those attached for 48 h or more. More than 60% of tick-bite victims removed adult ticks by 36 h of attachment, but only 10% found and removed the smaller nymphal ticks within the first 24 h of tick feeding. The duration of tick attachment may serve as a useful predictor of risk for acquiring various infections, such as Lyme disease and babesiosis, transmitted by I. scapularis. Regression equations developed herein correlate tick engorgement indices with duration of feeding. A table containing specific engorgement index prediction intervals calculated for both nymphs and adults will allow the practitioner or clinical laboratory to use easily measured tick engorgement indices to predict transmission risk by determining the duration of feeding by individual ticks.     

Effect of reduced bronchial circulation on lung fluid flux after smoke inhalation in sheep

Murine monoclonal antibodies directed against proteins of Borrelia burgdorferi B31 (low passage) were generated by the administration of antigen via the bite of borrelia-infected ticks. This strategy was employed as a mechanism to create antibodies against antigens presented by the natural route of tick transmission versus those presented by inoculation with cultured borreliae. One of the resultant antibodies reacted with a 17-kDa antigen from cultured B. burgdorferi, as seen by immunoblot analysis. This antibody was used to screen a B. burgdorferi genomic DNA lambda vector expression library, and an immunoreactive clone was isolated. DNA sequence analysis of this clone, containing a 2.7-kb insert, revealed several open reading frames. These open reading frames were found to be homologs of genes discovered as a multicopy gene family in the 297 strain of B. burgdorferi by Porcella et al. (S. F. Porcella, T. G. Popova, D. R. Akins, M. Li, J. D. Radolf, and M. V. Norgard, J. Bacteriol. 178:3293-3307, 1996). By selectively subcloning genes found in this insert into an Escherichia coli plasmid expression vector, the observation was made that the rev gene product was the protein reactive with the 17-kDa-specific monoclonal antibody. The rev gene product was found to be expressed in low-passage, but not in high-passage, B. burgdorferi B31. Correspondingly, the rev gene was not present in strain B31 genomic DNA from cultures that had been passaged >50 times. Serum samples from Lyme disease patients demonstrated an antibody response against the Rev protein. The generation of an anti-Rev response in Lyme disease patients, and in mice by tick bite inoculation, provides evidence that the Rev protein is expressed and immunogenic during the course of natural transmission and infection.     

Erythema migrans-like rash illness at a camp in North Carolina: a new tick-borne disease?

BACKGROUND: Borrelia burgdorferi, the causative agent of Lyme disease, has never been isolated from a patient thought to have acquired Lyme disease in any southeastern state. OBJECTIVE: To investigate 14 cases of an erythema migrans (EM)-like rash illness that occurred during 2 summers at an outdoor camp in central North Carolina in an effort to determine the etiologic, epidemiological, and clinical aspects of this illness. METHODS: Using active surveillance, we identified cases of clinically diagnosed EM in residents and staff of the camp. We collected clinical and demographic information; history of exposure to ticks; acute and convalescent serum antibodies to B. burgdorferi, Rickettsia rickettsii, and Ehrlichia chaffeensis; and cultures for spirochetes from biopsy specimens of skin lesions. Serum samples from a group of residents and staff who did not develop rashes were tested for the same antibodies. We speciated ticks removed from people and collected from vegetation. RESULTS: We identified 14 cases of EM-like rash illness during the 2 summers. Of the 14 case-patients, 10 had associated mild systemic symptoms and 1 had documented fever. All 14 case-patients had removed attached ticks, and 8 remembered having removed a tick from the site where the rash developed a median of 12 days earlier (range, 2-21 days). One tick removed from the site where a rash later developed was identified as Amblyomma americanum, the Lone Star tick; 97% of ticks collected from vegetation and 95% of ticks removed from people were A. americanum. No spirochetes were isolated from skin biopsy specimens. Paired serum samples from 13 case-patients did not show diagnostic antibody responses to B. burgdorferi or other tick-borne pathogens. CONCLUSIONS: This investigation suggests the existence of a new tick-associated rash illness. We suspect that the disease agent is carried by A. americanum ticks. In the southern United States, EM-like rash illness should no longer be considered definitive evidence of early Lyme disease.