Thomas J. Daniels

Host Feeding Patterns of Established and Potential Mosquito Vectors of West Nile Virus in the Eastern United States

An important variable in determining the vectorial capacity of mosquito species for arthropod-borne infections is the degree of contact of the vector and the vertebrate reservoir. This parameter can be estimated by examining the host-feeding habits of vectors. Serological and polymerase chain reaction based methods have been used to study the host-feedings patterns of 21 mosquito species from New York, New Jersey, and Tennessee, 19 of which previously have been found infected with West Nile virus. Mammalophilic mosquito species in New Jersey and New York fed primarily upon white-tailed deer, while those from Memphis, Tennessee, fed mainly upon domestic dogs. A total of 24 different avian host species were detected among the avian-derived blood meals. American Robin, Northern Cardinal, Northern Mockingbird, Tufted Titmouse, and Brown-headed Cowbird were common avian hosts, while blood meals derived from the American Crow were relatively rare. Although the majority of common host species were potentially among the most abundant birds at each location, the proportion of blood meals from the most commonly fed upon avian species was greater than was predicted based upon the likely abundance of these species alone. These findings suggest that vector species for West Nile virus may preferentially feed upon certain avian hosts.

Estimating Population Size and Drag Sampling Efficiency for the Blacklegged Tick (Acari: Ixodidae)

Abstract Estimates of absolute density were determined over a 5-yr period (1990–1994) for a population of Ixodes scapularis Say located in Westchester County, NY, by mark-release-recapture (nymphs and adults) and removal (larvae) methods. Density estimates for larvae ranged from 5.2 to 16.5/m2 and averaged 11.5/m2. Values for nymphs varied as much as fourfold among successive years, ranging from 0.5 to 2.3/m2 and averaging 1.2/m2, whereas adult density ranged from 0.3 to 0.4/m2, averaging 0.33/m2. Natural mortality of nymphs and adults was measured in experimental cages during population estimation periods, and indicated that survival declined linearly over the short-term and did not significantly influence estimates. Drag sampling efficiency, the proportion of the estimated population obtained in a single sample, averaged 6.3% among all stages. Efficiency was not significantly different among stages and was independent of tick density within a given life stage. The population estimation techniques employed in this study are well suited for use with I. scapularis and can provide data that offer insights into mortality patterns in individual populations.

Borrelia chilensis, a new member of the Borrelia burgdorferi sensu lato complex that extends the range of this genospecies in the Southern Hemisphere

Borrelia burgdorferi sensu lato (s.l.), transmitted by Ixodes spp. ticks, is the causative agent of Lyme disease. Although Ixodes spp. ticks are distributed in both Northern and Southern Hemispheres, evidence for the presence of B. burgdorferi s.l. in South America apart from Uruguay is lacking. We now report the presence of culturable spirochetes with flat-wave morphology and borrelial DNA in endemic Ixodes stilesi ticks collected in Chile from environmental vegetation and long-tailed rice rats (Oligoryzomys longicaudatus). Cultured spirochetes and borrelial DNA in ticks were characterized by multilocus sequence typing and by sequencing five other loci (16S and 23S ribosomal genes, 5S-23S intergenic spacer, flaB, ospC). Phylogenetic analysis placed this spirochete as a new genospecies within the Lyme borreliosis group. Its plasmid profile determined by polymerase chain reaction and pulsed-field gel electrophoresis differed from that of B. burgdorferi B31A3. We propose naming this new South American member of the Lyme borreliosis group B. chilensis VA1 in honor of its country of origin.

THE ROLE OF MEDIUM-SIZED MAMMALS AS RESERVOIRS OF BORRELIA BURGDORFERI IN SOUTHERN NEW YORK

The ability of raccoons (Procyon lotor), striped skunks (Mephitis mephitis) and opossums (Didelphis virginiana) to serve as reservoirs of Borrelia burgdorferi, the spirochetal agent of Lyme disease, was compared with that of white-footed mice (Peromyscus leucopus). Twenty-eight (28) medium-sized mammals and 34 white-footed mice were captured in Westchester County, New York (USA) in summer 1986. Animals were caged over pans of water for 1 to 2 days to recover engorged tick larvae (Ixodes dammini) that detached from the hosts after feeding. With the exception of mice, numbers of engorged tick larvae recovered exceeded those counted during initial examinations of the hosts by 30% (opossums) to nearly 90% (raccoons). Newly-molted nymphal ticks derived from the engorged larvae were examined for the presence of spirochetes by darkfield microscopy. Percentage infection was 5% (n = 22) for ticks from skunks and 14% (n = 191) for ticks from raccoons. None of 24 nymphs from larvae that fed on opossums survived long enough for spirochete examination. By comparison, 40% (n = 72) of nymphs from larvae which fed on white-footed mice were infected. Of the individual hosts from which molted nymphs had fed as larvae, 67% of mice, 33% of skunks, and 55% of raccoons produced spirochete-positive ticks.

The United States Department of Agriculture's Northeast Area-Wide Tick Control Project: Summary and Conclusions

From 1997 to 2002, the U.S. Department of Agricultures Northeast Area-wide Tick Control Project used acaricide-treated 4-Poster Deer Treatment Bait Stations in five eastern states to control ticks feeding on white-tailed deer. The objectives of this host-targeted technology were to reduce free-living blacklegged (Ixodes scapularis Say) and lone star (Amblyomma americanum [L.]) tick populations and thereby to reduce the risk of tick-borne disease. During 2002 to 2004, treatments were suspended, and tick population recovery rates were assayed. Subsequently, the major factors that influenced variations in efficacy were extrapolated to better understand and improve this technology. Treatments resulted in significant reductions in free-living populations of nymphal blacklegged ticks at six of the seven sites, and lone star ticks were significantly reduced at all three sites where they were present. During the study, maximal significant (p < or = 0.05) efficacies against nymphal blacklegged and lone star ticks at individual sites ranged from 60.0 to 81.7 and 90.9 to 99.5%, respectively. The major environmental factor that reduced efficacy was the occurrence of heavy acorn masts, which provided an alternative food resource for deer. Although the 4-Poster technology requires 1 or more years to show efficacy, this host-targeted intervention was demonstrated to be an efficacious, economical, safe, and environment-friendly alternative to area-wide spraying of acaricide to control free-living populations of these tick species.

Avian Reservoirs of the Agent of Human Granulocytic Ehrlichiosis

To the Editor: Human granulocytic ehrlichiosis (HGE), first described in 1994 (1), is the second-most common tick-borne disease in the United States; Lyme disease is the most prevalent. Both diseases are transmitted by blacklegged ticks (Ixodes scapularis Say) that are abundant in southern New York state (2). Factors that influence the risk for infection, particularly the role of wildlife in transmitting the etiologic agent, Anaplasma (formerly Ehrlichia) phagocytophilum (3,4), to vector ticks are not well understood. In the absence of transovarial transmission (5,6), acquisition of A. phagocytophilum by its vector must result either from feeding on the blood of reservoir-competent hosts or by cofeeding of uninfected ticks in close proximity to infected ticks (7).

Isolation of Entomopathogenic Fungi From Soils and Ixodes scapularis (Acari: Ixodidae) Ticks: Prevalence and Methods

ABSTRACT Entomopathogenic fungi are commonly found in forested soils that provide tick habitat, and many species are pathogenic to Ixodes scapularis Say, the blacklegged tick. As a first step to developing effective biocontrol strategies, the objective of this study was to determine the best methods to isolate entomopathogenic fungal species from field-collected samples of soils and ticks from an Eastern deciduous forest where I. scapularis is common. Several methods were assessed: (1) soils, leaf litter, and ticks were plated on two types of media; (2) soils were assayed for entomopathogenic fungi using the Galleria bait method; (3) DNA from internal transcribed spacer (ITS) regions of the nuclear ribosomal repeat was extracted from pure cultures obtained from soils, Galleria, and ticks and was amplified and sequenced; and (4) DNA was extracted directly from ticks, amplified, and sequenced. We conclude that (1) ticks encounter potentially entomopathogenic fungi more often in soil than in leaf litter, (2) many species of potentially entomopathogenic fungi found in the soil can readily be cultured, (3) the Galleria bait method is a sufficiently efficient method for isolation of these fungi from soils, and (4) although DNA extraction from ticks was not possible in this study because of small sample size, DNA extraction from fungi isolated from soils and from ticks was successful and provided clean sequences in 100 and 73% of samples, respectively. A combination of the above methods is clearly necessary for optimal characterization of entomopathogenic fungi associated with ticks in the environment.

Prevalence and Distribution of Ochlerotatus japonicus (Diptera: Culicidae) in Two Counties in Southern New York State

Abstract The seasonal occurrence and abundance of a newly introduced mosquito in the United States, Ochlerotatus (Finlaya) japonicus (Theobald), are reported for Westchester and Putnam Counties in southern New York State. Adult mosquitoes were sampled at 39 sites distributed throughout the two counties. CDC (Centers for Disease Control and Prevention) light traps (2,107 trap nights) and gravid traps (1,813 trap nights) were used to collect a total of 44,428 mosquitoes from May through October 2000. Oc. japonicus was found at 97.4% of sites sampled and accounted for 9.8% of all specimens collected. Oc. japonicus was recovered from 1.5% (n = 326) of the light trap collections and 18.1% (n = 4,026) of the gravid trap collections. Although gravid traps collected significantly more specimens than light traps, the seasonal activity patterns measured by each trap type were congruent. In all, 30 mosquito species were collected. Unlike other Ochlerotatus or Aedes species, Oc. japonicus was collected throughout the study, indicating a broad seasonal activity period. There was significant regional variation in Oc. japonicus abundance, with higher trap densities occurring in the northern-most trapping sites. This study demonstrates that Oc. japonicus is established in southern New York State.

Acaricidal treatment of white-tailed deer to control Ixodes scapularis (Acari: Ixodidae) in a New York Lyme disease-endemic community.

The efficacy of topically treating white-tailed deer with an acaricide was evaluated in a Lyme disease-endemic community of southern New York State. Twenty-four 4-Poster feeders were placed in a 5.2 km(2) treatment area in Bedford, NY, while a site in Lewisboro, NY, 4.8 km distant, served as control. Treatment periods ran from 15 September to 15 December each fall from 1997 to 2001, and from 15 March to 15 May each spring from 1998 to 2002. Corn consumption averaged 15,779 kg in fall sessions and 9054 kg in spring sessions, and a mean of 89.6% of deer in the study area showed evidence of using the feeders. Deer densities, estimated by aerial snow counts, averaged 22 and 28 deer per km(2) in Bedford and Lewisboro, respectively, over a 3-year period. Significant reductions in tick numbers on deer captured in the treatment area were noted in fall 1999 compared to deer captured at the control site. Drag sampling for nymphal host-seeking ticks indicated 63.6% control in 2001, which dropped to 54.8% the following year, but reached 80% in 2003. Higher-than-normal acorn production in 2001 that likely caused a drop in deer visitation to the feeders may have reduced efficacy against larval ticks in 2002. The 4-Poster effectively reduced the density of Ixodes scapularis, though the level of control is dependent on environmental factors that affect feeding behavior of white-tailed deer.

Microbiome changes through ontogeny of a tick pathogen vector

Blacklegged ticks (Ixodes scapularis) are one of the most important pathogen vectors in the United States, responsible for transmitting Lyme disease and other tick‐borne diseases. The structure of a hosts microbial community has the potential to affect the ecology and evolution of the host. We employed high‐throughput sequencing of the 16S rRNA gene V3‐V4 hypervariable regions in the first study to investigate the tick microbiome across all developmental stages (larvae, nymphs, adults). In addition to field‐collected life stages, newly hatched laboratory‐reared larvae were studied to determine the baseline microbial community structure and to assess transovarial transmission. We also targeted midguts and salivary glands due to their importance in pathogen maintenance and transmission. Over 100 000 sequences were produced per life stage replicate. Rickettsia was the most abundant bacterial genus across all sample types matching mostly the Ixodes rickettsial endosymbionts, and its proportion decreased as developmental stage progressed, with the exception of adult females that harboured a mean relative abundance of 97.9%. Laboratory‐reared larvae displayed the lowest bacterial diversity, containing almost exclusively Rickettsia. Many of the remaining bacteria included genera associated with soil, water and plants, suggesting environmental acquisition while off‐host. Female organs exhibited significantly different β‐diversity than the whole tick from which they were derived. Our results demonstrate clear differences in both α‐ and β‐diversity among tick developmental stages and between tick organs and the tick as a whole. Furthermore, field‐acquired bacteria appear to be very important to the overall internal bacterial community of this tick species, with influence from the host bloodmeal appearing limited.