Laboratory and Field Evaluations of a Commercially Available Real-Time Loop-Mediated Isothermal Amplification Assay for the Detection of West Nile Virus in Mosquito Pools.

Although the specific cDNA amplification mechanisms of reverse-transcriptase polymerase chain reaction (RT-PCR) and RT loop-mediated isothermal amplification (RT-LAMP) are very different, both molecular assays serve as options to detect arboviral RNA in mosquito pools. Like RT-PCR, RT-LAMP uses a reverse transcription step to synthesize complementary DNA (cDNA) from an RNA template and then uses target-specific primers to amplify cDNA to detectable levels in a single-tube reaction. Using laboratory-generated West Nile virus (WNV) samples and field-collected mosquito pools, we evaluated the sensitivity and specificity of a commercially available WNV real-time RT-LAMP assay (Pro-AmpRT™ WNV; Pro-Lab Diagnostics, Inc., Round Rock, Texas) and compared the results to a validated real-time RT-PCR assay. Laboratory generated virus stock samples containing ≥ 2.3 log10 plaque-forming units (PFU)/ml and intrathoracically inoculated mosquitoes containing ≥ 2.4 log10 PFU/ml produced positive results in the Pro-AmpRT WNV assay. Of field-collected pools that were WNV positive by real-time RT-PCR, 74.5% (70 of 94) were also positive by the Pro-AmpRT WNV assay, resulting in an overall Cohen's kappa agreement of 79.4% between the 2 tests. The Pro-AmpRT WNV assay shows promise as a suitable virus screening tool for vector surveillance programs provided agencies are aware of its characteristics and limitations.

Effect of Incandescent Light on Collection of West Nile Virus Vectors Using CDC Miniature Light Traps in Northern Colorado.

To evaluate whether the presence of clear incandescent light was attractive or refractive to host-seeking mosquitoes in northern Colorado, a Bayesian hierarchical model was created to measure differences in trap effectiveness based on presence or absence of phototactic cues. A total of eight CDC miniature light traps (with and without light) were set weekly across four locations in northern Colorado between Weeks 23 and 32 of year 2020. Culex mosquitoes (Diptera: Culicidae) accounted for 81% of all collections in this study with two vectors of West Nile virus being represented. The probability of catching both Culex tarsalis Coquillett and Culex pipiens Linnaeus was reduced when traps were equipped with light, but the difference was not statistically significant for Culex tarsalis. The clear reduction in the number of Culex pipiens caught when these traps were equipped with light indicates negative phototactic behavior and underestimation with current surveillance strategies. Removal of light from these traps may aid our understanding of these species' distribution within the environment, improve collection efficiency, and help guide implementation of targeted control measures used in public health mosquito control.

Preparations and Activities Necessary for Aerial Mosquito Control After Hurricanes.

Vector Disease Control International (VDCI) has a long history of aiding mosquito control efforts necessary for recovery after natural disasters like hurricanes and major floods. As waters associated with these events begin to recede, both nuisance and vector mosquito species surge in abundance and consequently play an increased role in public health. When these situations arise, state and county agencies implement emergency response plans and many rely on Federal Emergency Management Agency or private contractors for assistance in reducing mosquito populations that can alter arbovirus transmission cycles, cause intolerable stress, hamper reconstruction efforts, and disrupt normal community functions. Vector Disease Control International owns the largest fleet of fixed-wing aircraft dedicated specifically to mosquito control and has worked every major storm event since Hurricane Bonnie in 1998. This article describes the logistics and operations required for implementing VDCI's emergency management plan, including the relocation of equipment, adult mosquito surveillance, delivery of pesticides, assessment of efficacy, and filing of low-level waivers and congested-area plans with the Federal Aviation Administration.

Use of the vector index and geographic information system to prospectively inform West Nile virus interventions.

We sought to estimate West Nile virus (WNV) activity in mosquito populations weekly at the census tract level in Chicago, IL, and to provide this information graphically. Each week we calculated a vector index (VI) for each mosquito trap then generated tract estimates using geographic information systems. During June 29-September 13, 2008, a median of 527 (60%) of 874 possible tracts per week had a VI value. Overall, 94% of the weekly VI tract estimates were 0; among those with a VI estimate greater than 0, the median was 0.33 (range 0.003-3.5). Officials deemed risk levels and weather conditions appropriate for adulticide treatments on 3 occasions, resulting in the treatment of approximately 252 linear kilometers of residential streets and alleys. Our analysis successfully converted complex, raw surveillance data into a format that highlighted areas of elevated WNV activity and facilitated the determination of appropriate response procedures.

Passive tick surveillance, dog seropositivity, and incidence of human lyme disease.

Data on nymphal Ixodes scapularis ticks submitted by the public to the University of Rhode Island Tick Research Laboratory for testing from 1991 to 2000 were compared with human case data from the Rhode Island Department of Health to determine the efficacy of passive tick surveillance at assessing human risk of Lyme disease. Numbers of ticks submitted were highly correlated with human cases by county (r = 0.998, n = 5 counties) and by town (r = 0.916, n = 37 towns), as were the numbers of positive ticks submitted (r = 0.989 by county, r = 0.787 by town). Human cases were correlated with ticks submitted by town each year, and with positive ticks in all but 2 years. Thus, passive tick surveillance effectively assessed geographical risk of human Lyme disease. In contrast, tick submissions through time were not correlated with human cases from year to year. Dog seropositivity was significantly correlated with human cases by county in both years tested, but by town in only one of two years. Numbers of ticks submitted were correlated with dog seropositivity by county but not by town, apparently because of high variability among towns with small sample sizes. Our results suggest that passive tick surveillance, using ticks submitted by the public for Lyme spirochete testing, can be used to assess the geographical distribution of Lyme disease risk, but cannot reliably predict Lyme incidence from year to year.

Prevalence of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) in Ixodes scapularis (Acari: Ixodidae) adults in New Jersey, 2000-2001.

Using polymerase chain reaction, we analyzed 529 Ixodes scapularis Say adults collected from 16 of New Jersey's 21 counties for the presence of Borrelia burgdorferi, the etiological agent of Lyme disease. Overall, 261 (49.3%) were positive. B. burgdorferi was detected in ticks obtained from each county and from 53 of the 58 (93.1%) municipalities surveyed. The observed statewide prevalence in New Jersey is similar to those reported from other northeastern and mid-Atlantic states.