Cross-infectivity of Vorticella sp. across genera of mosquitoes for development of biological mosquito control strategies.

Protists in general comprise about one-third of the parasitic species infecting arthropod vectors, the role of free-living and epibiotic ciliates on mosquitoes have been insufficiently studied either due to their low pathogenicity or facultative parasites. Studies have shown that exposure of Paramecium ciliate protists, like Vorticella species, to first instar Culex nigripalpus Theobald, larvae delayed larval development and reduced biomass of emerged adults due to competition for food sources like bacteria and other microbes essential to mosquito growth and survival. Thus, we report on the capacity of a Vorticella sp. protist's ability to cross-infect host species and parasitize multiple mosquito larvae. The unique adapted behavior with the ability to remain on the exuviae in tree hole habitats provide a novel delivery system to develop products for target species-specific mosquitocides, larvicides, or viricides to be applied and sustained in aquatic systems.

Terrestrial Bird Migration and West Nile Virus Circulation, United States.

Host migration and emerging pathogens are strongly associated, especially with regard to zoonotic diseases. West Nile virus (WNV), a mosquitoborne pathogen capable of causing severe, sometimes fatal, neuroinvasive disease in humans, is maintained in highly mobile avian hosts. Using phylogeographic approaches, we investigated the relationship between WNV circulation in the United States and the flight paths of terrestrial birds. We demonstrated southward migration of WNV in the eastern flyway and northward migration in the central flyway, which is consistent with the looped flight paths of many terrestrial birds. We also identified 3 optimal locations for targeted WNV surveillance campaigns in the United States-Illinois, New York, and Texas. These results illustrate the value of multidisciplinary approaches to surveillance of infectious diseases, especially zoonotic diseases.

West Nile Virus Outbreak in Houston and Harris County, Texas, USA, 2014.

Since 2002, West Nile virus (WNV) has been detected every year in Houston and the surrounding Harris County, Texas. In 2014, the largest WNV outbreak to date occurred, comprising 139 cases and causing 2 deaths. Additionally, 1,286 WNV-positive mosquito pools were confirmed, the most reported in a single mosquito season.

Zika Virus Vector Competency of Mosquitoes, Gulf Coast, United States.

Zika virus has recently spread throughout the Americas. Although Aedes aegypti mosquitoes are considered the primary vector, Culex quinquefasciatus and mosquitoes of other species may also be vectors. We tested Cx. quinquefasciatus and Ae. taeniorhynchus mosquitoes from the US Gulf Coast; both were refractory to infection and incapable of transmission.

Efficacy of Deltagard®, Fyfanon®, and Evoluer™ 31-66 Against Culex quinquefasciatus In Harris County, Texas.

The comparative efficacy of ultra-low volume applications of deltamethrin, permethrin, and malathion against specific adult populations of Culex quinquefasciatus , a primary regional vector of Saint Louis encephalitis and West Nile virus, using field cage tests is described. At labeled application rates, the estimated mass median diameters for Fyfanon®, Evoluer™, and DeltaGard® were 18.32, 19.24, and 17.44 µm, respectively. No significant differences (P ≥ 0.01) in mean percent control were observed across all 3 wild populations and a susceptible Sebring strain tested with Fyfanon, DeltaGard, and Evoluer, providing 98.25%, 98.20%, and 97.24% mean percent control. Mean percent control for Sebring and 3 wild populations was 99.38%, 97.79%, 97.26%, and 97.15%, respectively, which were not significantly different from one another (P ≥ 0.01). Across all tests, highly significant differences (P ≤ 0.01) in mean percent control were obtained at 30.48, 60.96, and 91.44 m distances downwind, with mean percent control at 91.44 m being significantly less than that observed at 30.48 m, but not at 60.96 m. Individually, tests for DeltaGard, Fyfanon, and Evoluer resulted in no significant differences (P ≥ 0.01) in mean percent control between all mosquito populations tested, ranging from 96.47 to 100. However, Evoluer obtained significantly less (P ≤ 0.01) mean percent control at 91.44 m compared to DeltaGard and Fyfanon. Data suggest that the wild Cx. quinquefasciatus specimens collected were as susceptible as the laboratory Sebring strain. The use of Sebring as positive controls downwind strongly supported determination of the efficacy of all adulticides and susceptibility of wild populations tested.

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.

Fatigue from multiple host-seeking activity of Aedes albopictus and its effects on bloodfeeding behavior and deet repellency.

A decrease in host-seeking activity in female Aedes albopictus was observed after exposure to a human hand tested in an olfactometer in the laboratory. The increased treatment times resulted in a decreasing response to a human hand. Fatigued/exhausted female mosquitoes after multiple host-seeking activities did prolong the blood engorgement time, compared with untreated nonfatigued mosquitoes. Also, fatigued mosquitoes showed a significant reduction in human host-attacking rates. The mean duration of protection from mosquito bites of human hand treated with 25% deet (N,N-diethyl-3-methylbenzamide) was higher in fatigued than unfatigued mosquitoes.

First Report of Eratyrus cuspidatus (Hemiptera: Reduviidae) Infected with Trypanosoma cruzi in Peridomestic Environment in Chiapas, Mexico.

Background: Triatomine bugs are natural vectors of Trypanosoma cruzi, which causes Chagas disease or American trypanosomiasis. The role of sylvatic triatomine species as vectors of T. cruzi in Mexico remains to be fully understood. Our research on the epidemiology of Chagas disease in Southeastern Mexico involved sampling triatomines in rural settings. Materials and Methods: A triatomine was collected in a peridomestic environment of a rural dwelling in the state of Chiapas. The triatomine was identified morphologically as an adult female Eratyrus cuspidatus Stal. Results: Microscopic analysis revealed flagellate forms of T. cruzi in the feces of the E. cuspidatus collected. This was confirmed by quantitative polymerase chain reaction. Amplification of the mini-exon gene showed that the T. cruzi infecting E. cuspidatus corresponded to lineage I. Conclusions: This is the first report from Mexico of E. cuspidatus found infected in a human dwelling, which represents an important adaptation process to inhabit human environments.

Field evaluation of the Off! Clip-on Mosquito Repellent (metofluthrin) against Aedes albopictus and Aedes taeniorhynchus (Diptera: Culicidae) in northeastern Florida.

Repellent efficacy of the Off! Clip-on Mosquito Repellent device (S. C. Johnson and Son, Inc., Racine, WI) containing Metofluthrin was evaluated on six human volunteers against the container-breeding mosquito Aedes albopictus (Skuse) and the salt marsh mosquito Aedes taeniorhynchus (Wiedemann) at two field locations in northeastern Florida. The device repelled mosquitoes by releasing a vaporized form of the pyrethroid insecticide metofluthrin ([AI] 31.2%) and provided 70% protection from Ae. albopictus bites for > 3 h. For the second field trial, a repellent device that was used in the first trial was tested after being open for >1 wk. This device provided 79% protection from Ae. taeniorhynchus bites for 3 h. Our field results showed that the repellent device was 70 and 79% effective at repelling Ae. albopictus and Ae. taeniorhynchus from human test subjects in both field locations in northeastern Florida.

Survey of mosquito fauna in San Antonio, Texas.

A faunal survey of mosquitoes was conducted in 2008, 2009, and 2010 at various trapping locations throughout the city of San Antonio, TX. A total of 22,739 mosquitoes representing 41 species in 8 genera were collected during this study. The most prevalent mosquito species collected were Aedes vexans (25%), Ae. sollicitans (14%), Culex tarsalis (13.5%), and Cx. erraticus (11%). Four mosquito species (Ae. bimaculatus, Anopheles freeborni, Cx. declarator, and Psorophora horrida) not previously recorded in this area were collected, whereas several species historically occurring in San Antonio were not collected during this survey.

Ovicidal Aroma Shields for Prevention of Blow Fly Strikes Caused by Lucilia sericata (Meigen), Diptera: Calliphoridae.

The blow fly, Lucilia sericata (Meigen) (Diptera: Culicidae) is a primary facultative ectoparasite controlled by insecticides that have environmental and safety concerns; therefore, its natural and safe control is crucial. L. sericata eggs were subjected to 400 µL of 5% of 24 materials of plant-borne origin. The number of hatched and unhatched eggs were counted 24 h postexposure. Stopmyasis® expressed the highest ovicidal effect, followed by cedarwood, orange, and tea tree oils. The mean ovicidal results were 46.09-95.24% grouped as Class A, which provided the lowest hatchability rates. Even though benzoin, vanillin, citronella, and camphor oils grouped as Class B provided moderate ovicidal effects (33.69-43.92%), their efficacy differed significantly from those of the treatments in Classes A and C containing the control group and vetiver, eucalyptus, Olbas®, neem, sunflower, oil blends, patchouli, frankincense, p-menthane-3,8 diol (PMD), lavender, peppermint, cinnamon, calry sag, myrtle, and silicone oil. According to our knowledge, most (19 out of 24) applied materials were used as ovicides against L. serricata for the first time, except orange, eucalyptus, patchouli, cinnamon oils, and six of them were applied as ovicides against pests for the first time. Stopmyasis containing Géraniol and PMD is the drug of choice as an ecofriendly product to prevent blow fly strikes through correct diagnosis and prompt treatment preventing a disastrous and destructive course of the disease and improving quality of life.

Molecular analysis of knockdown resistance (kdr) mutations in the voltage-gated sodium channel gene of Aedes aegypti populations from Saudi Arabia.

BACKGROUND: The Aedes aegypti mosquito is the primary vector for dengue, chikungunya, yellow fever and Zika viruses worldwide. The first record of Ae. aegypti in southwestern Saudi Arabia was in 1956. However, the first outbreak and cases of dengue fever were reported in 1994, and cases have increased in recent years. Vector control for Ae. aegypti mainly uses pyrethroid insecticides in outdoor and indoor space spraying. The constant use of pyrethroids has exerted intense selection pressure for developing target-site mutations in the voltage-gated sodium channel (vgsc) gene in Ae. Aegypti against pyrethroids-mutations that have led to knockdown resistance (kdr). METHODS: Aedes aegypti field populations from five regions (Jazan, Sahil, Makkah, Jeddah and Madinah) of southwestern Saudi Arabia were genotyped for known kdr mutations in domains IIS6 and IIIS6 of the vgsc gene using polymerase chain reaction (PCR) amplification and sequencing. We estimated the frequency of kdr mutations and genotypes from Saudi Arabia as well as from other countries, Thailand, Myanmar (Southeast Asia) and Uganda (East Africa). We constructed haplotype networks to infer the evolutionary relationships of these gene regions. RESULTS: The three known kdr mutations, S989P, V1016G (IIS6) and F1534C (IIIS6), were detected in all five regions of Saudi Arabia. Interestingly, the triple homozygous wild genotype was reported for the first time in two individuals from the highlands of the Jazan region and one from the Al-Quoz, Sahil region. Overall, nine genotypes comprising four haplotypes were observed in southwestern Saudi Arabia. The median-joining haplotype networks of eight populations from Saudi Arabia, Southeast Asia and East Africa for both the IIS6 and IIIS6 domains revealed that haplotype diversity was highest in Uganda and in the Jazan and Sahil regions of Saudi Arabia, whereas haplotype diversity was low in the Jeddah, Makkah and Madinah regions. Median-joining haplotype networks of both domains indicated selection acting on the kdr-mutation containing haplotypes in Saudi Arabia. CONCLUSIONS: The presence of wild type haplotypes without any of the three kdr mutations, i.e. that are fully susceptible, in Saudi Arabia indicates that further consideration should be given to insecticide resistance management strategies that could restore pyrethroid sensitivity to the populations of Ae. aegypti in Saudi Arabia as part of an integrative vector control strategy.

Insecticide resistance in bedbugs in Thailand and laboratory evaluation of insecticides for the control of Cimex hemipterus and Cimex lectularius (Hemiptera: Cimicidae).

Bedbugs are found in many countries around the world, and in some regions they are resistant to numerous insecticides. This study surveyed bedbugs in Thailand and determined their resistance to insecticides. The surveys were carried out in six provinces that attract large numbers of foreign tourists: Bangkok, Chonburi, Chiang Mai, Ubon Ratchathani, Phuket, and Krabi. Bedbugs were collected from hotels and colonized in the laboratory to evaluate their resistance to insecticides. Cimex hemipterus (F.) was found in some hotels in Bangkok, Chonburi, Phuket, and Krabi, whereas Cimex lectularius L. was found only in hotels in Chiang Mai. No bedbugs were found in Ubon Ratchathani. The colonized bedbugs showed resistance to groups of insecticides, including organochlorines (dichlorodiphenyl trichloroethane, dieldrin), carbamates (bendiocarb, propoxur), organophosphates (malathion, fenitrothion), and pyrethroids (cyfluthrin, deltamethrin, permethrin, lambda-cyhalothrin, etofenprox) in tests using World Health Organization insecticide-impregnated papers. The new insecticides imidacloprid (neonicotinoid group), chlorfenapyr (pyrrole group), and fipronil (phenylpyrazole group) were effective against the bedbugs; however, organophosphate (diazinon), carbamates (fenobucarb, propoxur), and pyrethroids (bifenthrin, cypermethrin, esfenvalerate, etofenprox) were ineffective. Aerosols containing various pyrethroid insecticides with two to four different active ingredients were effective against the bedbugs. The results obtained from this study suggested that both species of bedbugs in Thailand have developed marked resistance to various groups of insecticides, especially those in the pyrethroid group, which are the most common insecticides used for pest control. Therefore, an integrated pest management should be implemented for managing bedbugs in Thailand.

Kdr genotyping (V1016I, F1534C) of the Nav channel of Aedes aegypti (L.) mosquito populations in Harris County (Houston), Texas, USA, after Permanone 31-66 field tests and its influence on probability of survival.

Aedes aegypti (L.) is an important mosquito vector of emerging arboviruses such as Zika, dengue, yellow fever, and chikungunya. To quell potential disease outbreaks, its populations are controlled by applying pyrethroid insecticides, which selection pressure may lead to the development of insecticide resistance. Target site insensitivity to pyrethroids caused by non-synonymous knockdown resistance (kdr) mutations in the voltage-gated sodium (NaV) channel is a predominant mechanism of resistance in mosquitoes. To evaluate the potential impact of pyrethroid resistance on vector control, Ae. aegypti eggs were collected from eight mosquito control operational areas in Harris County, Texas, and emerged females were treated in field tests at four different distances from the pyrethroid Permanone 31-66 source. The females were genotyped by melting curve analyses to detect two kdr mutations (V1016I and F1534C) in the NaV channel. Harris County females had higher survivorship rates at each distance than the pyrethroid-susceptible Orlando strain females. Survivorship increased with distance from the pyrethroid source, with 39% of field-collected mosquitoes surviving at 7.62 m and 82.3% at 22.86 m from the treatment source. Both the V1016I and F1534C pyrethroid resistant genotypes were widely distributed and at high frequency, with 77% of the females being double homozygous resistant (II/CC), this being the first report of kdr mutations in Ae. aegypti in Harris County. Analysis of the probability of survival for each mutation site independently indicated that the CC genotype had similar probability of survival as the FC heterozygous, while the II genotype had higher survival than both the VI and VV, that did not differ. The double homozygous resistant genotype (II/CC) had the highest probability of survival. A linear model estimated probability of survival for areas and genotypes. The high frequency and widespread distribution of double-homozygote pyrethroid-resistant Ae. aegypti may jeopardize disease vector control efforts in Harris County.

Evaluation of a Novel User-Friendly Arthropod Repellent Gel, Verdegen.

Hand sanitizers are developed as alcohol-based liquid gel formulations, generally used to decrease the amount of infectious agents on human hands. Verdegen, LLC proposed to prepare an arthropod repellent gel for public use when the recent outbreaks of Zika infection vectored through Aedes mosquitoes in the American continents prompted multi-faceted emergency measures. Four different gel formulations were developed, comprising two of the most efficacious commercial arthropod repellent active ingredients, N,N-diethyl-3-methyl benzamide (deet) and 2-(2-hydroxyethyl)-1-piperidinecarboxylic acid 1-methylpropyl ester (picaridin), each at different concentrations (20 and 33% deet, or 20 and 33% picaridin). Compliance with the use of topical arthropod repellents remains an issue among military personnel. One of the most common complaints by Soldiers is that they do not like how the repellents applied on their skin leave behind an oily or greasy residue. These new gel formulations offer a user-friendly alternative for commonly used arthropod repellents formulations for the military and civilian personnel. We tested the efficacy and protection time of these new gel formulations in comparison with the commercially available cream formulations of deet and picaridin at similar concentrations. Our data show that gel formulations have better topical attributes, and offer equal or better biting protection for up to 48 h against host-seeking Aedes aegypti (L.) (Diptera: Culicidae) female mosquitoes.

Characterisation of actions of p-menthane-3,8-diol repellent formulations against Aedes aegypti mosquitoes.

BACKGROUND: Characterisation of mosquito repellents using arm-in-cage tests are performed by assessing the 95% effective dose (ED95), half-life and complete protection time (CPT). This study fully characterizes these properties for p-menthane-3,8-diol (PMD), which has not been widely studied, and a long-acting formulation containing a PMD-vanillin composite. METHODS: A series of arm-in-cage tests against Aedes aegypti (Diptera: Culicidae) mosquitoes were devised using 6 volunteers to estimate CPT or 10 to estimate the ED95 and half-lives for three repellents: 20% N,N-diethyl-3-methylbenzamide (DEET), 30% PMD and a novel 30% PMD-vanillin formulation. Non-linear regression analysis was used to characterize the relationship between applied dose and CPT. RESULTS: PMD and DEET showed a very similar log dose relationship to CPT; however, the PMD-vanillin formulation exhibited a sigmoidal 'S-shaped' relationship. This resulted in a 1.5-fold higher CPT for PMD-vanillin compared with that of 20% DEET when applied at a dose of 1.6 mg/cm2, but little difference was observed at lower doses of 0.8-1 mg/cm2. The ED95 value for the 30% PMD and PMD-vanillin formulations were 0.25 and 0.24 mg/cm2, respectively, these being higher than that for 20% DEET (0.09 mg/cm2). The half-lives for 30% PMD and 20% DEET were similar (2.23 vs. 2.74 h), but longer for the PMD-vanillin formulations (3.8 h). CONCLUSIONS: A full characterisation for other repellent formulations, particularly those claiming extended longevity, should be conducted in order to identify differences at various applied doses.

Detection of the Nav channel kdr-like mutation and modeling of factors affecting survivorship of Culex quinquefasciatus mosquitoes from six areas of Harris County (Houston), Texas, after permethrin field-cage tests.

Culex quinquefasciatus is one of the most important mosquito vectors of arboviruses. Currently, the fastest approach to control disease transmission is the application of synthetic adulticide insecticides. However, in highly populated urban centers the development of insecticide resistance in mosquito populations could impair insecticide efficacy and therefore, disease control. To assess the effect of resistance on vector control, females of Cx. quinquefasciatus collected from six mosquito control operational areas in Harris County, Texas, were treated in field cage tests at three different distances with the pyrethroid Permanone® 31-66 applied at the operational rate. Females were analyzed by sequencing and/or diagnostic PCR using de novo designed primers for detecting the kdr-like mutation in the voltage-gated sodium channel (L982F; TTA to TTT) (house fly kdr canonical mutation L1014F). Females from the Cx. quinquefasciatus susceptible Sebring strain and those from the six operational areas placed at 30.4 m from the treatment source were killed in the tests, while 14% of field-collected mosquitoes survived at 60.8 m, and 35% at 91.2 m from the source. The diagnostic PCR had a with 97.5% accuracy to detect the kdr-like mutation. Pyrethroid resistant mosquitoes carrying the L982F mutation were broadly distributed in Harris County at high frequency. Among mosquitoes analyzed (n = 1,028), the kdr-kdr genotype was prevalent (81.2%), the kdr-s genotype was 18%, and s-s mosquitoes were less than 1% (n = 8). A logistic regression model estimated an equal probability of survival for the genotypes kdr-kdr and kdr-s in all areas analyzed. Altogether, our results point to a high-risk situation for the pyrethroid-based arboviral disease control in Harris County.

Isolongifolenone: a novel sesquiterpene repellent of ticks and mosquitoes.

A naturally occurring sesquiterpene, isolongifolenone, derivatives of which have been used extensively as ingredients in the cosmetics industry, was discovered to effectively repel blood-feeding arthropods that are important disease vectors. We show that (-)-isolongifolenone deters the biting of the mosquitoes, Aedes aegypti (L.) and Anopheles stephensi Liston, more effectively than the widely used synthetic chemical repellent, N,N-diethyl-3-methyl benzamide (DEET), in laboratory bioassays. The compound also repelled blacklegged ticks, Ixodes scapularis Say, and lone star ticks, Amblyomma americanum (L.), as effectively as DEET. Isolongifolenone is easily synthesized from inexpensive turpentine oil feedstock. We are therefore confident that the compound has significant potential as an inexpensive and safe repellent for protection of large human populations against blood-feeding arthropods.

Attractiveness of botanical infusions to ovipositing Culex quinquefasciatus, Cx. nigripalpus, and Cx. erraticus in San Antonio, Texas.

Field experiments were conducted on the Fort Sam Houston Military Reservation, San Antonio, TX, in fall 2008 to observe the attractiveness of selected botanical infusions to ovipositing female mosquitoes. The following infusions were tested in Centers for Disease Control and Prevention gravid traps: Bermuda grass (Cynodon dactylon), oak leaf (Quercus virginiana), acacia leaf (Acacia schaffneri), rabbit chow (alfalfa pellets), and algae (Spirogyra sp.). Four (Bermuda, acacia, oak, and algae) of the 5 infusions were effective in collecting Culex quinquefasciatus, Cx. nigripalpus, and Cx. erraticus. Of the 4 infusions, Bermuda collected the greatest number of the mosquitoes sampled. Female Aedes albopictus mosquitoes were collected in moderate numbers during this study.

Laboratory and field evaluation of SS220 and deet against mosquitoes in Queensland, Australia.

Laboratory and field efficacy trials comparing deet (N,N-diethyl-3-methylbenzamide) and SS220 [(IS, 2'S)-2-methylpiperindinyl-3-cyclohexen-1-carboxamide] against mosquitoes in Queensland, Australia, were conducted. In the laboratory, both compounds provided between 150 and 195 min of protection against Aedes aegypti and between 18 and 80 min of protection against Anopheles farauti. In laboratory tests against Culex annulirostris, 20% SS220 provided 3 h of protection and 20% deet provided >6 h of protection. A field efficacy test was conducted at Redcliffe, Queensland in January 2008 and the predominant mosquito species collected was Cx. annulirostris (84.4% of collection). In the field, 20% SS220 provided significantly better protection against mosquitoes than 20% deet. Seven hours after application, SS220 provided greater than 96.0% protection against all mosquitoes, whereas 20% deet provided 58.9% protection.