Efficacy of Two Larvasonic™ Units Against Culex Larvae and Effects on Common Aquatic Nontarget Organisms in Harris County, Texas.

The Larvasonic™ Field Arm Mobile Wetlands Unit and SD-Mini were tested for efficacy against Culex larvae, and effects on aquatic nontarget organisms (NTO). The Field Arm provided 84.61% to 100% control of caged Culex larvae out to 0.91-m distance in shallow ditches and 60.45% control of Culex larvae at 0.61-m without any effects to caged NTO. Slow ditch treatment achieved 77.35% control compared to fast treatment (20.42%), whereas 77.65% control was obtained along edges of a neglected swimming pool, compared to near the middle (23.97%). In bucket tests, the SD-Mini provided >97% control of Culex and 85.35% reduction of immature giant water bugs, which decreased slightly (83.45%) over the monitoring period, which was not significantly different from cannibalistic damselflies (62.80%), with reduction of both being significantly higher than other NTO tested. There was a small (0.37%) reduction of dragonflies (naiads), due to cannibalism. Both Larvasonic units could effectively augment conventional larvicide operations in smaller areas without causing resistance within mosquito populations or harming NTO when used properly.

Continued evolution of West Nile virus, Houston, Texas, USA, 2002-2012.

We investigated the genetics and evolution of West Nile virus (WNV) since initial detection in the United States in 1999 on the basis of continual surveillance studies in the Houston, Texas, USA, metropolitan area (Harris County) as a surrogate model for WNV evolution on a national scale. Full-length genomic sequencing of 14 novel 2010-2012 WNV isolates collected from resident birds in Harris County demonstrates emergence of 4 independent genetic groups distinct from historical strains circulating in the greater Houston region since 2002. Phylogenetic and geospatial analyses of the 2012 WNV isolates indicate closer genetic relationship with 2003-2006 Harris County isolates than more recent 2007-2011 isolates. Inferred monophyletic relationships of these groups with several 2006-2009 northeastern US isolates supports potential introduction of a novel WNV strain in Texas since 2010. These results emphasize the need to maintain WNV surveillance activities to better understand WNV transmission dynamics in the United States.

Changing socioeconomic indicators of human plague, New Mexico, USA.

Socioeconomic indicators associated with temporal changes in the distribution of human plague cases in New Mexico were investigated for 1976-2007. In the 1980s, cases were more likely in census block groups with poor housing conditions, but by the 2000s, cases were associated with affluent areas concentrated in the Santa Fe-Albuquerque region.

Evolution of new genotype of West Nile virus in North America.

Previous studies of North American isolates of West Nile virus (WNV) during 1999-2005 suggested that the virus had reached genetic homeostasis in North America. However, genomic sequencing of WNV isolates from Harris County, Texas, during 2002-2009 suggests that this is not the case. Three new genetic groups have been identified in Texas since 2005. Spread of the southwestern US genotype (SW/WN03) from the Arizona/Colorado/northern Mexico region to California, Illinois, New Mexico, New York, North Dakota, and the Texas Gulf Coast demonstrates continued evolution of WNV. Thus, WNV continues to evolve in North America, as demonstrated by selection of this new genotype. Continued surveillance of the virus is essential as it continues to evolve in the New World.

Thermal fog efficacy tests against mosquitoes in storm drains in Harris County, Texas 2008-09.

In 2008 and 2009, our current thermal fogging methods and materials were evaluated in underground systems against feral and laboratory strains of adult Culex quinquefasciatus. Culex quinquefasciatus adults collected from storm drain systems the night before treatment were tested concurrently with adults from a susceptible laboratory strain (Sebring) in 10 separate tests. During 2008, there were no significant differences in the low mean percent control obtained between any of the feral populations (29.39%) and susceptible (Sebring) populations (56.04%) tested, whereas in 2009, application of mineral oil alone yielded extremely low, but significantly different mean percent control at 0.99% for ferals and 0.01% for Sebrings. During 2009 mineral oil evaluations, 45,677 droplets were collected in storm drains at distances of 99.1, 50.6, 57.9, 67.7, and 109.7 m from the application site, with 99% of the droplets below 3 microm in diam; additionally, we found no significant differences between mean percent control of Sebring and feral mosquito populations using the higher (3.2x) 1:10 application rate of Pyrocide. However, mean percent control between the feral and susceptible strain (Sebring) during 2009 was lower than in 2008 at 16.55% for ferals and 24.43% in Sebrings. Results indicated that control methodologies and/or chemicals used were ineffective at controlling Cx. quinquefasciatus in storm drains using the chosen experimental design. Based upon this information, thermal fog operations were discontinued due to lack of effectiveness.

A comparison of gravid and under-house CO2-baited CDC light traps for mosquito species of public health importance in Houston, Texas.

The relative efficacy of gravid and under-house CO2 traps for monitoring mosquito species of public health importance within the Houston metroplex area was assessed. Gravid and under-house traps were colocated at 10 sites and monitored weekly between 1 March to 31 May 2007. The most numerous species caught was Culex pipiens quinquefasciatus Say. Other species of public health importance caught in gravid and under-house traps included Culex restuans Theobald, Aedes aegypti (L.), and Aedes albopictus Skuse. Adjusting for the week of collection, gravid traps caught significantly more mosquitoes (mean 23.1 per trap) in the study area than under-house traps (mean 3.6 per trap). However, under-house traps caught a greater variety of mosquito species (13) than gravid traps (11). Gravid and under-house traps only caught nine of 15 of the same mosquito species during the study period. In this study area, gravid traps should be used as the primary method of surveillance for mosquito-borne diseases of public health importance during the early part of the season, because of greater catch numbers of mosquitoes that pose a public health risk.

The influence of weather and weather variability on mosquito abundance and infection with West Nile virus in Harris County, Texas, USA.

Early warning systems for vector-borne diseases (VBDs) prediction are an ecological application where data from the interface of several environmental components can be used to predict future VBD transmission. In general, models for early warning systems only consider average environmental conditions ignoring variation in weather variables, despite the prediction from Schmalhausen's law about the importance of environmental variability for biological systems. We present results from a long-term mosquito surveillance program from Harris County, Texas, USA, where we use time series analysis techniques to study the abundance and West Nile virus (WNV) infection patterns in the local primary vector, Culex quinquefasciatus Say. We found that, as predicted by Schmalhausen's law, mosquito abundance was associated with the standard deviation and kurtosis of environmental variables. By contrast, WNV infection rates were associated with 8-month lagged temperature, suggesting environmental conditions during overwintering might be key for WNV amplification during summer outbreaks. Finally, model validation showed that seasonal autoregressive models successfully predicted mosquito WNV infection rates up to 2 months ahead, but did rather poorly at predicting mosquito abundance, a result that might reflect impacts of vector control for mosquito population reduction, geographic scale, and other artifacts generated by operational constraints of mosquito surveillance systems.

Characterization of Three Novel Viruses from the Families Nyamiviridae, Orthomyxoviridae, and Peribunyaviridae, Isolated from Dead Birds Collected during West Nile Virus Surveillance in Harris County, Texas.

This report describes and characterizes three novel RNA viruses isolated from dead birds collected during West Nile virus surveillance in Harris County, TX, USA (the Houston metropolitan area). The novel viruses are identified as members of the families Nyamaviridae, Orthomyxoviridae, and Peribunyaviridae and have been designated as San Jacinto virus, Mason Creek virus, and Buffalo Bayou virus, respectively. Their potential public health and/or veterinary importance are still unknown.

Biological and phylogenetic characterization of pigeon paramyxovirus serotype 1 circulating in wild North American pigeons and doves.

As part of West Nile virus surveillance programs in Rhode Island and eastern Texas between 2000 and 2007, brain tissue was collected from 5,608 dead birds representing 21 avian orders found in public places or reported by homeowners. Fifteen Newcastle disease virus isolates were recovered only from birds of the order Columbiformes and were positively identified by the USDA-validated real-time reverse transcription-PCR assay targeting the matrix gene and more specifically as pigeon paramyxovirus serotype 1 (PPMV-1) by hemagglutinin inhibition with monoclonal antibodies. Based upon partial genomic sequencing and phylogenetic analysis, the newly isolated viruses represent a distinct sublineage within class II genotype VIb. All of the viruses (15/15) were classified as virulent based upon their fusion cleavage site motif ((112)RRKKRF(117)) and intracerebral pathogenicity indices of >0.7 (ranging from 0.98 to 1.35); however, these viruses escaped detection by the fusion gene-based real-time PCR test for virulence. Modifications introduced to the probe site of the fusion gene-based assay allowed rapid virulence detection within this distinct sublineage.

A multiunit CO2 anesthetizing system for use in transferring mosquitoes during field cage insectcide efficacy tests.

A multiunit CO2 delivery and anesthetizing system that supports expanded cage testing of mosquitoes is described. This system has proven more efficient than pipetting for conducting multiple tests and has facilitated the tripling of cage testing capabilities.

Residence-linked human plague in New Mexico: a habitat-suitability model.

Yersinia pestis, the causative agent of plague, has been detected in fleas and mammals throughout the western United States. This highly virulent infection is rare in humans, surveillance of the disease is expensive, and it often was assumed that risk of exposure to Y. pestis is high in most of the western United States. For these reasons, some local health departments in these plague-affected regions have hesitated to undertake surveillance and other prevention activities. To aid in targeting limited public health resources, we created a fine-resolution human plague risk map for New Mexico, the state reporting more than half the human cases in the United States. Our GIS-based model included three landscape features-a nonlinear relationship with elevation, distance to water, and distance to the ecotone between Rocky Mountain/Great Basin open and closed coniferous woodlands-and yielded an overall accuracy of approximately 80%. The model classified 17.25% of the state as posing significant risk of exposure to humans on privately or tribally owned land, which suggests that resource requirements for regular surveillance and control of plague could be effectively focused on < 20% of the state.

Host feeding pattern of Culex quinquefasciatus (Diptera: Culicidae) and its role in transmission of West Nile virus in Harris County, Texas.

The vertebrate hosts of 672 blood-engorged Culex quinquefasciatus Say, collected in Harris County, Texas, during 2005, were identified by nucleotide sequencing PCR products of the cytochrome b gene. Analysis revealed that 39.1% had acquired blood from birds, 52.5% from mammals, and 8.3% were mixed avian and mammalian blood meals. Most frequent vertebrate hosts were dog (41.0%), mourning dove (18.3%), domestic cat (8.8%), white-winged dove (4.3%), house sparrow (3.2%), house finch (3.0%), gray catbird (3.0%), and American robin (2.5%). Results are interpreted in conjunction with concurrent avian and mosquito West Nile virus (WNV) surveillance activities in Harris County. We conclude that Cx. quinquefasciatus is an opportunistic feeder and principal mosquito vector of WNV in this metropolitan area; however, transmission by other mosquito species or by other modes of infection, such as ingestion, must account for the high WNV infection rates among local blue jays and American crows.

Human plague in the southwestern United States, 1957-2004: spatial models of elevated risk of human exposure to Yersinia pestis.

Plague is a rare but highly virulent flea-borne zoonotic disease caused by the Gram-negative bacterium Yersinia pestis Yersin. Identifying areas at high risk of human exposure to the etiological agent of plague could provide a useful tool for targeting limited public health resources and reduce the likelihood of misdiagnosis by raising awareness of the disease. We created logistic regression models to identify landscape features associated with areas where humans have acquired plague from 1957 to 2004 in the four-corners region of the United States (Arizona, Colorado, New Mexico, and Utah), and we extrapolated those models within a geographical information system to predict where plague cases are likely to occur within the southwestern United States disease focus. The probability of an area being classified as high-risk plague habitat increased with elevation up to approximately 2300 m and declined as elevation increased thereafter, and declined with distance from key habitat types (e.g., southern Rocky Mountain piñon--juniper [Pinus edulis Engelm. and Juniperus spp.], Colorado plateau piñon--juniper woodland, Rocky Mountain ponderosa pine (Pinus ponderosa P.& C. Lawson var. scopulorum), and southern Rocky Mountain juniper woodland and savanna). The overall accuracy of the model was >82%. Our most conservative model predicted that 14.4% of the four-corners region represented a high risk of peridomestic exposure to Y. pestis.

A multiple cage-holding, wind-sensitive vane design for use in ground adulticiding efficacy testing in Harris County, Texas.

A wind-sensitive vane apparatus was designed and implemented specifically to accommodate the attachment of otherwise unidirectional insecticide treatment cages used in ground ultra-low volume mosquito adulticide field tests. This cage support system is useful in keeping the potential West Nile virus and St. Louis encephalitis vector Culex quinquefasciatus caged mosquitoes oriented into the wind during field efficacy tests. Testing capacity for resistance surveillance was tripled during the 2005 season, and more reliable results were achieved as a consequence.

Description and use of the Harris County gravid trap for West Nile virus surveillance 2003-06.

The Harris County Gravid Trap (HCGT) used for West Nile virus (WNV) surveillance from 2003-06 is described. The HCGT collected 1,918,483 Culex quinquefasciatus females (over 36,627 periods), with 466,296.37 (8,452.74) yearly. Only 48,094 Aedes albopictus females (over 15,139 periods) were collected, with 10,277.79 (3,260.33) yearly. The highest mosquito production occurred in July for Cx. quinquefasciatus (63,851.25 +/- 0.13), and for Ae. albopictus (2,981.80 +/- 0.09). Integrating gravid traps within our program has increased our ability to detect WNV efficiently in both species.

Urban habitat evaluation for West Nile virus surveillance in mosquitoes in Albuquerque, New Mexico.

As part of an ongoing mosquito surveillance program, 27 sites in the greater metropolitan Albuquerque area (Bernalillo County, New Mexico) were trapped from May through September 2004. Each site was sampled for 1 night weekly, using a standard CO2-baited Centers for Disease Control and Prevention light trap and a gravid trap. Captured mosquitoes were catalogued by location, species, and date, and selected pools were tested for West Nile virus (WNV) by reverse transcription-polymerase chain reaction. Based on previous surveillance, WNV was already established in the state of New Mexico. Surveillance during 2003, the 1st year of WNV detection in New Mexico mosquitoes, was focused on the bosque forest of the Rio Grande river valley. Surveillance during summer of 2004 was extended to additional areas around the city of Albuquerque, the state's largest population center. In addition to the standard surveillance objectives, a secondary goal was to determine whether foci of WNV activity were detectable in other habitats besides the riparian ecosystem of the Rio Grande, and in other species not previously identified as vectors. There was no demonstrable advantage to extending the traditional trapping area outside of the Rio Grande valley. Sites in the valley area had WNV-positive mosquitoes earlier in the season, and for a longer period than the added sites. In addition, riparian sites had the highest diversity of species, the largest numbers of Culex spp. captured, and the largest proportion of the WNV-positive mosquito pools from the study. Species found in other areas of the metropolitan area were also represented in the valley. Although WNV activity was detected in other areas of the city, its activity began later and ended earlier than in the river valley. We surmise that the greatest benefit to mosquito surveillance could be achieved by focusing on the river valley area.

Associations between two mosquito populations and West Nile virus in Harris County, Texas, 2003-06.

Associations between Culex quinquefasciatus, Aedes albopictus and West Nile virus (WNV) activity, temperature, and rainfall in Harris County, Texas 2003-06 are discussed. Human cases were highly correlated to Cx. quinquefasciatus (r = 0.87) and Ae. albopictus (r = 0.78) pools, blue jays (r = 0.83), and Ae. albopictus collected (r = 0.71), but not Cx. quinquefasciatus collected (r = 0.45). Human cases were associated with temperature (r = 0.71), not rainfall (r = 0.29), whereas temperature correlated with Ae. albopictus and Cx. quinquefasciatus collections (r = 0.88 and 0.70, respectively) and Cx. quinqueftsciatus pools (r = 0.75), but not Ae. albopictus pools (r = 0.55). Both species (collections and pools) and blue jays were weakly correlated (r 5 0.41) with rainfall, but blue jays were better correlated with Cx. quinquefasciatus pools (r = 0.87), compared with Ae. albopictus pools (r = 0.67), Ae. albopictus collections (r = 0.69), and Cx. quinquefasciatus collections (r = 0.46). Peak minimum infection rate for Cx. quinquefasciatus (4.55), and Ae. albopictus (4.41) was in August with highest human cases (17.87), blue jays (55.58), and temperature (29.01 degrees C). Between both species, blood meal analysis indicated 68.18% of Cx. quinquefasciatus mammalian hosts were dog, while 22.72% were human, whereas Ae. albopictus had higher human (44.44%) but fewer dog hosts (22.22%). Ten bird species were identified as hosts for Cx. quinquefasciatus, with northern cardinal and blue jay representing 26.66% and 20.00%, respectively. No bird feeding activity was observed in Ae. albopictus. The earliest and latest human blood meal occurred in May (Ae. albopictus) and November (Cx. quinquefasciatus); 66.66% of human host identifications between both species occurred in October-November, after the seasonal human case peak. Based upon our data, WNV activity in both mosquito species warrants further investigation of their individual roles in WNV ecology within this region.

Emergence of West Nile virus in mosquito (Diptera: Culicidae) communities of the New Mexico Rio Grande Valley.

The first appearances of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in New Mexico were reported in late summer to early fall 2002. Several dead birds tested positive for WNV, and 78 equine cases were confirmed. All mosquito pools tested (n = 268) were negative. A statewide surveillance program was launched in May 2003 to study the emergence and spread of this new arbovirus in mosquitoes from the Rio Grande valley. Mosquitoes were trapped at 32 sites along a 750-km stretch of the Rio Grande valley. Sites were trapped for one night either weekly or biweekly, by using CO2-baited CDC light traps and gravid traps. Pools of captured mosquitoes were tested for WNV by reverse transcription-polymerase chain reaction. By mid-July 2003, WNV levels in the mosquito population had reached levels that were detectable by the surveillance program. Positive pools of mosquitoes were found in the Rio Grande valley from mid-July through late September. In total, 75 positive pools were found, from sites throughout the study area. The predominant species infected with WNV in this region were Culex tarsalis (Coquillett) in rural areas, and Culex salinarius (Coquillett) and Culex pipiens quinquefasciatus (Say) in urban areas. There were 202 human cases and 438 equine cases of WNV in New Mexico in 2003, which corresponded well in time with the positive mosquitoes. Our results seemed to be consistent with introduction of WNV in late summer 2002, followed by a period of transmission and amplification cycles between local avian hosts and mosquito vectors.

Estimation of aerosol droplet sizes by using a modified DC-III portable droplet measurement system under laboratory and field conditions.

Modification of the DC-III portable droplet measurement system, permitting its use under field conditions, is described. Under laboratory conditions, the system effectively sampled water droplets from aerosols produced by a dry ice/water generator and high-pressure syringe. Seven droplet sizes, totaling 71,053 droplets within 22 tests (dry ice method), consisted of 1-, 2-, 6-, 11-, 18-, 25-, and 34-microm droplets with individual (rounded) percentages of 45.25, 37.22, 13.85, 3.17, 0.45, 0.02, and 0.005, respectively, for each size. Cumulatively, 1-microm droplets accounted for ca. 45.25% of the droplets sampled; combined with 2-microm (ca. 82.48% together), 6-microm (ca. 96.33% together), and 11-microm droplets, yielded ca. 99.51% of the droplets sampled. The syringe produced 12 droplet sizes, with 4,121 droplets sampled, consisting of 1, 2, 6, 11, 18, 25, 34, 45, 56, 69, 83, and 99 microm with individual percentages of 15.43, 21.91, 24.58, 17.30, 10.62, 4.65, 2.93, 1.33, 0.63, 0.33, 0.16, 0.07, respectively, for each size. The 6-microm droplets contributed the highest individual percentage, and cumulatively, these droplets combined with 1- and 2-microm droplets, yielding 61.93%, whereas 11- to 45-microm droplets contributed 36.83%, for a total of 98.76%. Droplets measuring 56-99 microm accounted for ca. 1.24% of droplets sampled. Hand-fogger oil aerosols produced 12 droplet sizes (1-38 microm) at test distances of 7.6 and 15.2 m, with 1,979 and 268 droplets sampled, respectively, during 10 tests at each distance. With analysis of variance of transformed individual percentages for each size at both distances, no significant differences were observed for 7.6 and 15.2 m. Cumulatively, 1-, 2-, 3-, and 5-microm droplets contributed 82.87 and 80.97%, whereas 8-, 11-, 14-, and 18-microm droplets added 14.55% to totals at both 7.6 and 15.2 m, respectively. Droplets measuring 22, 27, 32, and 38 microm contributed 2.57% and 4.47% to samples obtained at 7.6 and 15.2 m. The average mass median diameter (MMD) of mineral oil aerosols obtained at 7.6 and 15.2 m were 19.55 +/- 1.62 and 15.49 +/- 1.35, respectively. Positioned at 15 m (50 ft) within a field cage test plot during 12 separate tests, the MMD values obtained downrange were less than calibration MMD values in 5 tests (10-93% of calibration values), but 1.1 to 4.4 times greater in 7 remaining tests. With exception of 2 tests, susceptible Sebring strain Culex quinquefasciatus mortality at the 31-m (100-ft) stake, 15 m (50 ft) behind the DC-III, was consistently 100%. The average MMD values for malathion, permethrin, and resmethrin (at 0.003 and 0.007 rates) at this distance were 13.24 +/- 6.08, 7.48 +/- 5.69, 16.64 +/- 5.54, and 15.48 +/- 2.89, respectively.

Comparison of mosquito trapping method efficacy for West Nile virus surveillance in New Mexico.

As part of the West Nile virus surveillance program for the state of New Mexico, 13 sites along the Rio Grande River were sampled for mosquitoes during spring and summer 2003. We evaluated 3 different trapping procedures for their effectiveness at capturing selected species of mosquitoes. The 3 methods used were a dry ice-baited Centers for Disease Control and Prevention (CDC) light trap set 1.5 m above the ground (standard method), a CDC light trap suspended within the forest canopy, and a gravid trap set on the ground. Thirteen sites were sampled for 10 1-night periods biweekly from May through September. The relative numbers of captured Culex tarsalis, Cx. salinarius, Cx. quinquefasciatus, and Aedes vexans as well as the numbers of total recorded captures of all species were compared for each trapping method. Significant differences were observed for each species by location and by trapping method. Culex tarsalis was most commonly caught in canopy or standard CDC traps, especially in cottonwood bosque. Culex salinarius was found most frequently in association with marshy water, and was most often caught in gravid or standard light traps. Culex quinquefasciatus was captured almost exclusively in gravid traps within urban areas. Aedes vexans was primarily sampled in standard CDC light traps and found most frequently in wooded areas near floodplains. With the exception of Cx. Quinquefasciatus, no species was collected significantly more frequently in gravid or canopy traps than in the standard CDC light trap. Our findings do not support altering the methods currently used in New Mexico, namely, the use of 1.5-m CDC light traps and gravid traps. An increased use of gravid traps seems to be warranted in monitoring urban vector populations (specifically Cx. quinquefasciatus and Cx. salinarius) that may be involved in human transmission.