Comparison of the efficiency and cost of West Nile virus surveillance methods in California.

Surveillance systems for West Nile virus (WNV) combine several methods to determine the location and timing of viral amplification. The value of each surveillance method must be measured against its efficiency and costs to optimize integrated vector management and suppress WNV transmission to the human population. Here we extend previous comparisons of WNV surveillance methods by equitably comparing the most common methods after standardization on the basis of spatial sampling density and costs, and by estimating optimal levels of sampling effort for mosquito traps and sentinel chicken flocks. In general, testing for evidence of viral RNA in mosquitoes and public-reported dead birds resulted in detection of WNV approximately 2-5 weeks earlier than serological monitoring of sentinel chickens at equal spatial sampling density. For a fixed cost, testing of dead birds reported by the public was found to be the most cost effective of the methods, yielding the highest number of positive results per $1000. Increased spatial density of mosquito trapping was associated with more precise estimates of WNV infection prevalence in mosquitoes. Our findings also suggested that the most common chicken flock size of 10 birds could be reduced to six to seven without substantial reductions in timeliness or sensitivity. We conclude that a surveillance system that uses the testing of dead birds reported by the public complemented by strategically timed mosquito and chicken sampling as agency resources allow would detect viral activity efficiently in terms of effort and costs, so long as susceptible bird species that experience a high mortality rate from infection with WNV, such as corvids, are present in the area.

Surveillance for Ixodes pacificus and the tick-borne pathogens Anaplasma phagocytophilum and Borrelia burgdorferi in birds from California's Inner Coast Range.

We investigated the involvement of birds in the ecology of the western black-legged tick, Ixodes pacificus, and its associated zoonotic bacteria, Borrelia burgdorferi and Anaplasma phagocytophilum, at two interior coast-range study sites in northern California. Anaplasma phagocytophilum, the agent of granulocytic anaplasmosis (GA), and B. burgdorferi s.s., the agent of Lyme disease (LD), are tick-borne pathogens that are well established in California. We screened blood and ticks from 349 individual birds in 48 species collected in 2011 and 2012 using pathogen-specific PCR. A total of 617 immature I. pacificus was collected with almost three times as many larvae than nymphs. There were 7.5 times more I. pacificus at the Napa County site compared to the Yolo County site. Two of 74 (3%) nymphal pools from an Oregon junco (Junco hyemalis) and a hermit thrush (Catharus guttatus) and 4 individual larvae (all from Oregon juncos) were PCR-positive for B. burgdorferi. Blood samples from a golden-crowned sparrow (Zonotrichia atricapilla) and a European starling (Sturnus vulgaris) were positive for A. phagocytophilum DNA at very low levels. Birds that forage on ground or bark and nest on the ground, as well as some migratory species, are at an increased risk for acquiring I. pacificus. Our findings show that birds contribute to the ecologies of LD and GA in California by serving as a blood-meal source, feeding and transporting immature I. pacificus, and sometimes as a source of Borrelia infection.

Phenotypic variation among Culex pipiens complex (Diptera: Culicidae) populations from the Sacramento Valley, California: horizontal and vertical transmission of West Nile virus, diapause potential, autogeny, and host selection.

The vector competence and bionomics of Culex pipiens form pipiens L. and Cx. pipiens f. molestus Forskäl were evaluated for populations from the Sacramento Valley. Both f. pipiens and f. molestus females became infected, produced disseminated infections, and were able to transmit West Nile virus. Form molestus females also transmitted West Nile virus vertically to egg rafts and F1 progeny, whereas f. pipiens females only transmitted to egg rafts. Culex pipiens complex from urban Sacramento blood-fed on seven different avian species and two mammalian species. Structure analysis of blood-fed mosquitoes identified K = 4 genetic clusters: f. molestus, f. pipiens, a group of genetically similar hybrids (Cluster X), and admixed individuals. When females were exposed as larvae to midwinter conditions in bioenvironmental chambers, 85% (N = 79) of aboveground Cx. pipiens complex females and 100% (N = 34) of underground f. molestus females did not enter reproductive diapause.

Overwintering biology of Culex (Diptera: Culicidae) mosquitoes in the Sacramento Valley of California.

At temperate latitudes, Culex (Diptera: Culicidae) mosquitoes typically overwinter as adult females in reproductive arrest and also may serve as reservoir hosts for arboviruses when cold temperatures arrest viral replication. To evaluate their role in the persistence of West Nile virus (WNV) in the Sacramento Valley of California, the induction and termination of diapause were investigated for members of the Culex pipiens (L.) complex, Culex tarsalis Coquillett, and Culex stigmatosoma Dyar under field, seminatural, and experimental conditions. All Culex spp. remained vagile throughout winter, enabling the collection of 3,174 females and 1,706 males from diverse habitats during the winters of 2010-2012. Overwintering strategies included both quiescence and diapause. In addition, Cx. pipiens form molestus Forskäl females remained reproductively active in both underground and aboveground habitats. Some blood-fed, gravid, and parous Cx. tarsalis and Cx. pipiens complex females were collected throughout the winter period. Under both field and experimental conditions, Cx. tarsalis and Cx. stigmatosoma females exposed to autumnal conditions arrested primary follicular maturation at previtellogenic stage I, with primary to secondary follicular ratios <1.5 (indicative of a hormonally induced diapause). In contrast, most Cx. pipiens complex females did not enter reproductive diapause and ovarian follicles matured to >or=stage I-II (host-seeking arrest) or were found in various stages of degeneration. Diapause was initiated in the majority of Cx. tarsalis and Cx. stigmatosoma females by mid-late October and was terminated after the winter solstice, but host-seeking seemed limited by temperature. An accrual of 97.52 +/- 30.7 and 162.85 +/- 79.3 degree-days after the winter solstice was estimated to be necessary for diapause termination in Cx. tarsalis under field and seminatural conditions, respectively. An increase in the proportion of blood-fed Culex females in resting collections occurred concurrently with diapause termination in field populations based on ovarian morphometrics. WNV RNA was detected in one pool of 18 males and in a single blood-fed female Cx. tarsalis collected during winter. Therefore, both vertically and horizontally infected Culex females may persist through winter and possibly transmit WNV after diapause termination in late winter or early spring in the Sacramento Valley of California.

Evaluation of efficacy and human health risk of aerial ultra-low volume applications of pyrethrins and piperonyl butoxide for adult mosquito management in response to West Nile virus activity in Sacramento County, California.

The Sacramento and Yolo Mosquito and Vector Control District (SYMVCD, also referred to as "the District") conducts surveillance and management of mosquitoes in Sacramento and Yolo counties in California. Following an increase in numbers and West Nile virus (WNV) infection rates of Culex tarsalis and Culex pipiens, the District decided on July 26, 2007, to conduct aerial applications of Evergreen EC 60-6 (60% pyrethrins: 6% piperonyl butoxide) over approximately 215 km2 in the north area of Sacramento County on the nights of July 30, July 31, and August 1, 2007. At the same time, the District received notification of the first human WNV case in the area. To evaluate the efficacy of the applications in decreasing mosquito abundance and infection rates, we conducted pre- and post-trapping inside and outside the spray zone and assessed human health risks from exposure to the insecticide applications. Results showed a significant decrease in abundance of both Cx. tarsalis and Cx. pipiens, and in the minimum infection rate of Cx. tarsalis. Human-health risks from exposure to the insecticide were below thresholds set by the US Environmental Protection Agency.

A probabilistic risk assessment for deployed military personnel after the implementation of the "Leishmaniasis Control Program" at Tallil Air Base, Iraq.

Leishmaniasis has been of concern to the U.S. military and has re-emerged in importance because of recent deployments to the Middle East. We conducted a retrospective probabilistic risk assessment for military personnel potentially exposed to insecticides during the "Leishmaniasis Control Plan" (LCP) undertaken in 2003 at Tallil Air Base, Iraq. We estimated acute and subchronic risks from resmethrin, malathion, piperonyl butoxide (PBO), and pyrethrins applied using a truck-mounted ultra-low-volume (ULV) sprayer and lambda-cyhalothrin, cyfluthrin, bifenthrin, chlorpyrifos, and cypermethrin used for residual sprays. We used the risk quotient (RQ) method for our risk assessment (estimated environmental exposure/toxic endpoint) and set the RQ level of concern (LOC) at 1.0. Acute RQs for truck-mounted ULV and residual sprays ranged from 0.00007 to 33.3 at the 95th percentile. Acute exposure to lambda-cyhalothrin, bifenthrin, and chlorpyrifos exceeded the RQ LOC. Subchronic RQs for truck-mounted ULV and residual sprays ranged from 0.00008 to 32.8 at the 95th percentile. Subchronic exposures to lambda-cyhalothrin and chlorpyrifos exceeded the LOC. However, estimated exposures to lambda-cyhalothrin, bifenthrin, and chlorpyrifos did not exceed their respective no observed adverse effect levels.

Environmental concentrations, fate, and risk assessment of pyrethrins and piperonyl butoxide after aerial ultralow-volume applications for adult mosquito management.

One of the most effective ways of managing adult mosquitoes that vector human and animals diseases is the use of ultralow-volume insecticides. Because of concerns about the safety of the insecticides used for the management of adult mosquitoes, we conducted an environmental fate and efficacy study in Princeton and Colusa (both CA, USA) after aerial applications of pyrethrins and piperonyl butoxide (PBO). One hour before application, PBO concentrations in water were 0.008 and 0.2175 microg/L for Princeton and Colusa, respectively. One hour after the spray event in Princeton, the average PBO concentrations were 0.0125 microg/cm2 on ground-deposition pads and 0.1723 microg/L in water samples, with concentrations decreasing significantly over time. One hour after the spray event in Colusa, the average PBO concentrations were 0.0199 microg/cm2 on deposition pads and 1.274 microg/L in water samples, with concentrations decreasing significantly over time. A significant time and location effect for both deposition pads and water samples in Princeton and Colusa was observed (p<0.001 and p=0.014, respectively). Pyrethrins were not detected in nearly all ground and water samples. One hour after application, mortality of Culex tarsalis and Culex pipiens in sentinel cages was significantly higher than at the control site for both locations (p<0.001). Risk quotients for aquatic surrogate species in Princeton and Colusa were 0.002 or less at 1 h after application, which did not exceed the U.S. Environmental Protection Agency risk quotient level of concern for endangered aquatic organisms of 0.05. Our results suggest that the amounts of pyrethrins and PBO deposited on the ground and in water after aerial ULV insecticide applications are lower than those estimated by previous exposure and risk assessments.

Risk assessments for exposure of deployed military personnel to insecticides and personal protective measures used for disease-vector management.

Infectious diseases are problematic for deployed military forces throughout the world, and, historically, more military service days have been lost to insect-vectored diseases than to combat. Because of the limitations in efficacy and availability of both vaccines and therapeutic drugs, vector management often is the best tool that military personnel have against most vector-borne pathogens. However, the use of insecticides may raise concerns about the safety of their effects on the health of the military personnel exposed to them. Therefore, our objective was to use risk assessment methodologies to evaluate health risks to deployed U.S. military personnel from vector management tactics. Our conservative tier-1, quantitative risk assessment focused on acute, subchronic, and chronic exposures and cancer risks to military personnel after insecticide application and use of personal protective measures in different scenarios. Exposures were estimated for every scenario, chemical, and pathway. Acute, subchronic, and chronic risks were assessed using a margin of exposure (MOE) approach. Our MOE was the ratio of a no-observed-adverse-effect level (NOAEL) to an estimated exposure. MOEs were greater than the levels of concern (LOCs) for all surface residual and indoor space spraying exposures, except acute dermal exposure to lambda-cyhalothrin. MOEs were greater than the LOCs for all chemicals in the truck-mounted ultra-low-volume (ULV) exposure scenario. The aggregate cancer risk for permethrin exceeded 1 x 10(-6), but more realistic exposure refinements would reduce the cancer risk below that value. Overall, results indicate that health risks from exposures to insecticides and personal protective measures used by military personnel are low.

An ecological risk assessment for insecticides used in adult mosquito management.

West Nile virus (WNV) has been a concern for people across the United States since the disease was initially observed in the summer of 1999. Since 1999, WNV has caused the largest arboviral encephalitis epidemic in US history. Vector control management programs have been intensively implemented to control mosquitoes that carry WNV. Our deterministic ecological risk assessment focused on 6 common mosquito adulticides used in vector control, including 3 pyrethroids (d-phenothrin, resmethrin, and permethrin), pyrethrins, and 2 organophosphates (malathion and naled). Piperonyl butoxide, a synergist for the pyrethroids, was also assessed. Both aquatic and terrestrial nontarget organisms were considered for acute and chronic exposures to the adulticides. Tier I exposure estimates were derived from ISCST3 and AERMOD for deposition and air concentrations affecting terrestrial organisms and PRZM-EXAMS for standard pond concentrations affecting aquatic organisms. Nontargets exposed to adulticides included small mammals, birds, as well as aquatic vertebrates and invertebrates in a pond subject to receiving the chemical via drift and runoff. Risk quotients were obtained by comparing exposures to toxic endpoints. All risk quotients were low indicating that risks to ecological receptors most likely were small.

A human-health risk assessment for West Nile virus and insecticides used in mosquito management.

West Nile virus (WNV) has been a major public health concern in North America since 1999, when the first outbreak in the Western Hemisphere occurred in New York City. As a result of this ongoing disease outbreak, management of mosquitoes that vector WNV throughout the United States and Canada has necessitated using insecticides in areas where they traditionally have not been used or have been used less frequently. This has resulted in concerns by the public about the risks from insecticide use. The objective of this study was to use reasonable worst-case risk assessment methodologies to evaluate human-health risks for WNV and the insecticides most commonly used to control adult mosquitoes. We evaluated documented health effects from WNV infection and determined potential population risks based on reported frequencies. We determined potential acute (1-day) and subchronic (90-day) multiroute residential exposures from each insecticide for several human subgroups during a WNV disease outbreak scenario. We then compared potential insecticide exposures to toxicologic and regulatory effect levels. Risk quotients (RQs, the ratio of exposure to toxicologic effect) were < 1.0 for all subgroups. Acute RQs ranged from 0.0004 to 0.4726, and subchronic RQs ranged from 0.00014 to 0.2074. Results from our risk assessment and the current weight of scientific evidence indicate that human-health risks from residential exposure to mosquito insecticides are low and are not likely to exceed levels of concern. Further, our results indicate that, based on human-health criteria, the risks from WNV exceed the risks from exposure to mosquito insecticides.