The effect of feeding duration on pathogen transmission was studied for individual ticks infected with either laboratory or field strains of the Lyme disease spirochete Borrelia burgdorferi and field strains of Ehrlichia phagocytophila, an agent of human granulocytic ehrlichiosis. Infected nymphal Ixodes scapularis were allowed to feed individually on mice, and equal numbers were removed at 24-h intervals for < or =96 h. Mice were assayed for infection by culture, serologic testing, and polymerase chain reaction (PCR) analysis. Fed ticks were assayed by culture or PCR analysis. Transmission of B. burgdorferi did not occur during the first 24 h among 66 attempts, with maximum transmission occurring between 48 and 72 h. A model estimating the probability of infection from individual ticks removed by patients in a Lyme disease-endemic area yielded an overall probability of 4.6%. Infected I. scapularis nymphs transmitted E. phagocytophila within 24 h in 2 of 3 attempts, which indicates that daily tick removal may not be adequate to prevent human infection with this agent.

Primers were used to amplify a 561-bp region of the 16S rRNA gene of Ehrlichia phagocytophila from Ixodes scapularis ticks and small mammals collected in Rhode Island and Connecticut. DNA sequences for all 50 E. phagocytophila-positive samples collected from 1996 through 1998 in southwestern Connecticut were identical to the sequence reported for E. phagocytophila DNA from confirmed human cases. In contrast, the sequences from 92 of 123 E. phagocytophila-positive Rhode Island samples collected from 1996 through 1999 included several variants differing by 1-2 nucleotides from that in the agent infecting humans. While 11.9% of 67 E. phagocytophila-positive ticks collected during 1997 in Rhode Island harbored ehrlichiae with sequences identical to that of the human agent, 79.1% had a variant sequence not previously described. The low incidence of human ehrlichiosis in Rhode Island may in part result from interference by these variant ehrlichiae with maintenance and transmission of the true agent of human disease.

Survival of unfed larvae and nymphs of Ixodes scapularis Say (formerly named I. dammini Spielman, Clifford, Piesman & Corwin) was tested at 27 degrees C for relative humidities ranging from 65 to 100%, which were maintained by various saturated salts or water. The times until half the larvae died (LT50) at 100, 93, 85, 75, and 65% RH were 67.1, 26.6, 8.3, 1.3, and 1.1 d, respectively. Maximal larval survival in the laboratory was 129, 59, 24, 3, and 2 d at the same relative humidities. There was a distinct difference between the mortality curves for larvae at 93% RH and 100% RH, which suggests that larvae were taking in water from the air at the higher RH whereas those at an RH < or = 93% were not. There was little difference between the 100 and 93% RH survival curves for the nymphs. All of the nymphs at 100% RH survived at least 149 d. At 93 and 100% RH, one nymph survived at each RH for 210 and 205 d, respectively. The LT50s for the nymphs of I. scapularis were 169.9 (100% RH), 168.5 (93%), 118.8 (85%), 10.7 (75%), and 3.6 (65%) d. Maximal nymphal survival in the laboratory was 162, 65, and 8 d at 85, 75, and 65% RH, respectively. The inability of the larvae to retain water effectively is probably more important than the ability to imbibe water from subsaturated air in the survival of this stage. Subadult I. scapularis require a relatively humid habitat for survival, although some individuals have the potential to seek a host for many weeks within a mesic environment.

A 3-yr community-based study was conducted on residential properties on Mason's Island, Mystic, CT, to determine the efficacy of a rodent-targeted acaricide (fipronil) to control immature Ixodes scapularis (Say) on Peromyscus leucopus. Results indicated that modified commercial bait boxes were effective as an acaricide delivery method for reducing nymphal and larval tick infestations on white-footed mice by 68 and 84%, respectively. Passive application of fipronil significantly reduced the infection rate of Borrelia burgdorferi among white-footed mice by 53%. Moreover, the abundance of questing I. scapularis adults on treated properties was reduced by 77% and fewer were infected with spirochetes (31%) compared with untreated sites (47%) after 3 yr of treatment. Likewise, the abundance of host-seeking nymphs was significantly reduced on treated properties by >50%. Finally, infection rates in flagged nymphal ticks for both B. burgdorferi and Anaplasma phagocytophilum were reduced by 67 and 64%, respectively, after only 2 yr of treatment. Results from this 3-yr trial indicate that the use of fipronil passively applied to reservoir animals by bait boxes is an environmentally acceptable means to control ticks, interrupt the natural disease transmission cycle, and reduce the risk of Lyme disease for residents of treated properties.

Serum samples from Peromyscus leucopus (white-footed mouse), collected in Connecticut (USA) in 1983, 1985, and during 1990 to 1993, were analyzed by an enzyme-linked immunosorbent assay (ELISA) or indirect fluorescent antibody (IFA) staining methods for antibodies to Borrelia burgdorferi (strain 2591), Babesia microti, Ehrlichia chaffeensis (Arkansas strain), and Ehrlichia equi (MRK strain). Of the 294 serum samples tested, 160 (54%) contained immunoglobulins to one or more of these pathogens. There were antibodies to two or more etiologic agents in 77 (48%) of the seropositive mice. Although it was uncommon to detect coexisting antibodies to all four pathogens (n = 5 positive mice), E. chaffeensis-reactive antibodies or immunoglobulins to E. equi were present along with those produced to B. burgdorferi and B. microti in 24 other mice. These rodents carry antibodies to several tick-borne pathogens at numerous sites in Connecticut and may play a role in the epizootiology of ehrlichiosis as well as babesiosis and Lyme borreliosis.

White-tailed deer (Odocoileus virginianus Zimmerman), serve as the primary host for the adult blacklegged tick (Ixodes scapularis Say), the vector for Lyme disease, human babesiosis, and human granulocytic anaplasmosis. Our objective was to evaluate the degree of association between deer density, tick abundance, and human cases of Lyme disease in one Connecticut community over a 13-yr period. We surveyed 90-98% of all permanent residents in the community six times from 1995 to 2008 to document resident's exposure to tick-related disease and frequency and abundance of deer observations. After hunts were initiated, number and frequency of deer observations in the community were greatly reduced as were resident-reported cases of Lyme disease. Number of resident-reported cases of Lyme disease per 100 households was strongly correlated to deer density in the community. Reducing deer density to 5.1 deer per square kilometer resulted in a 76% reduction in tick abundance, 70% reduction in the entomological risk index, and 80% reduction in resident-reported cases of Lyme disease in the community from before to after a hunt was initiated.

The principal vector for the pathogens of Lyme disease, human granulocytic ehrlichiosis, and human babesiosis is the tick Ixodes scapularis Say. A chalcid wasp, Ixodiphagus hookeri, in the family Encyrtidae parasitizes populations of the tick on several islands or other geographically isolated sites in New England with high densities of these ticks and white-tailed deer (Odocoileus virginianus), the principal host for adult I. scapularis. Deer densities were reduced at a forested tract in Bridgeport and the Bluff Point Coastal Reserve in Groton, Connecticut, from levels exceeding 90 animals per km2 in 1992 (Bridgeport) and 1994 (Bluff Point) to 17 and 10 animals per km2, respectively, by fall 2001. Tick densities declined with sustained reductions in the population of white-tailed deer. Similarly, prevalence of tick parasitism by Ixodes hookeri declined at both sites from 30 to 25% to <1.0% and was significantly correlated with previous year's deer density at both sites (r(s) = 0.933 and r(s) = 0.867, P < or = 0.0001) and with nymphal tick densities at Bridgeport (r(s) = 0.867, P < or = 0.0001), but was not as well correlated with tick densities in Groton. The virtual disappearance of I. hookeri in this study corresponds with a lack of I. hookeri in mainland I. scapularis at comparable deer and tick densities, suggesting that there is a threshold deer density of approximatley 10-20/km2, with corresponding tick densities necessary for I. hookeri to successfully parasitize I. scapularis.

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