Novel effective mosquito larvicide DL-methionine: Lack of toxicity to non-target aquatic organisms.

Mosquito larvicides are an effective tool for reducing numbers of adult females that bite and potentially spread pathogenic organisms. Methionine, an essential amino acid in humans, has been previously demonstrated to be a highly effective larvicide against four (4) mosquito species in three (3) genera, Anopheles, Culex and Aedes. The aim of the present study was to determine the potential impact on non-target aquatic organisms of methionine applied as a mosquito larvicide. DL-methionine concentrations ranging from 0.06% to 1.00% were used; wherein the highest concentration of 1.00% would result in 100% mortality within 48 h in mosquitoes. Acute toxicity assays were conducted in accordance with the US Environmental Protection Agency (US EPA) guidelines for the water flea (Daphnia magna Straus; Cladocera: Daphniidae) and the fathead minnow (Pimephales promelas Rafinesque; Cypriniformes: Cyprinidae). Water fleas and fish were placed directly into the solutions in glass containers and tanks for 48-hours and 96-hours, respectively. When applied within the above-mentioned range of effective mosquito larvicide concentrations, DL-methionine meets US EPA criteria as a "practically non-toxic" pesticide for both species. These results suggest that methionine is a viable alternative to current mosquito larvicide options, which are typically classified as moderately to highly toxic and may be a valuable addition to a mosquito integrated pest management program.

Combining DL-Methionine and Bacillus thuringiensis Subspecies israelensis: Prospects for a Mosquito Larvicide.

Mosquito larvicides can reduce mosquito populations at the source, potentially decreasing biting rates and pathogen transmission. However, there is a growing need for mosquito larvicides that are environmentally sustainable. Bacillus thuringiensis subspecies israelensis (BTI) is a naturally occurring bacterium commonly used as a larvicide to manage mosquito populations. Methionine is an essential amino acid that has demonstrated toxic properties against larval mosquitoes in laboratory experiments, while having minimal effects on non-target organisms. The goal of this study was to evaluate the potential for a novel combination larvicide by testing for compatibility between these two active ingredients. We began by determining the lethal concentration values (LCs) of BTI and DL-methionine against Anopheles quadrimaculatus Say and Aedes aegypti Linnaeus (Diptera: Culicidae) larvae. These bioassays were conducted in glass jars and mortality was observed 48 h post-treatment. We found that while DL-methionine was more toxic to An. quadrimaculatus than Ae. aegypti, the opposite was true for BTI. Then, we used these LCs to conduct bioassays with a combination of BTI and DL-methionine to determine the relationship between the two active ingredients when used against An. quadrimaculatus and Ae. aegypti larvae. The findings of this study demonstrate that BTI and DL-methionine have the potential to be complementary due to their additive properties at higher concentrations and effect levels when tested against An. quadrimaculatus. However, an antagonistic relationship was detected at the concentrations tested with Ae. aegypti. These results are encouraging and imply that a DL-methionine or BTI/DL-methionine combination larvicide could be used in management of Anopheles species.

Can the Addition of Odor and Visual Targets Enhance Attraction of the Asian Citrus Psyllid (Hemiptera: Liviidae) to Sticky Traps?

Asian citrus psyllid, Diaphorina citri Kuwayama, negatively impacts the citrus industry as it transmits Candidatus Liberibacter spp., the causal agent of citrus greening. Monitoring D. citri population levels is critical for management of vectors and citrus greening and is usually through use of yellow sticky traps. In our study, use of odors, odor blends, and visual targets were evaluated to determine whether attraction to yellow sticky traps could be improved. Methyl salicylate consistently increased D. citri attraction to decoy yellow but not to other yellow or yellow/green traps. Addition of a visual target did not enhance attraction to sticky traps. While several chemical blends were evaluated, they did not increase psyllid attraction to decoy yellow traps. The increased attraction to traps with methyl salicylate is promising and may contribute to trapping efficiency under field conditions.

Evaluation of Fipronil Susceptibility in the Lone Star Tick (Acari: Ixodidae).

The lone star tick, Amblyomma americanum Linnaeus, is ubiquitously present in the southeastern United States and will readily parasitize humans and companion animals. Fipronil is the active ingredient in several topically applied products used to manage ticks and fleas on companion animals. Reducing ectoparasite infestations on companion animals decreases the risk that pathogens vectored by these pests are transmitted to these animals and their owners. However, dependence on acaricides can promote the development of resistance. In this study, the Food and Agriculture Organization of the United Nations larval packet test was used to determine the lethal concentration (LC) values and discriminating concentration (DC) for fipronil in the lone star tick. The DC was calculated as 0.02080%. The low magnitude of the DC value suggests that fipronil is an effective active ingredient for A. americanum management. With the LC and DC values determined, emergent resistance can be monitored, potentially allowing for intervention should tolerance develop in A. americanum populations that are in frequent contact with humans and their companion animals.

Mortality Effects of Three Bacterial Pathogens and Beauveria bassiana When Topically Applied or Injected Into House Flies (Diptera: Muscidae).

The house fly, Musca domestica L., is a global pest of public health and agricultural importance. The efficacy of conventional management has been waning due to increasing insecticide resistance. A potential management tool is the entomopathogenic fungus, Beauveria bassiana Vuillemin (Hypocreales: Cordycipitaceae) (strain L90), although time-to-death is slower than desired by potential users. This research investigated the effectiveness of three gram-negative bacteria (Pseudomonas protegens Ramette (Psuedomonadales: Pseudomonadaceae) pf-5, Photorhabdus temperata Fischer-Le Saux (Enterobacteriales: Enterobacteriaceae) NC19, and Serratia marcescens Bizio (Enterobacteriales: Enterobacteriaceae) DB11) on house fly mortality when topically applied, compared to B. bassiana. Each pathogen's virulence was measured by injection into adult female house flies or by topical applications to their thorax. All bacterial strains were highly virulent after injection with 1 × 104 colony forming units (cfu), causing fly mortality within 24 h. Beauveria bassiana resulted in high mortality, 3 d postinjection at the high dose of 1 × 104 conidia/µl. Mortality due to topical treatments of P. temperata and S. marcescens was low even at the highest dose of 1 × 106 cfu/µl. Mortality after topical treatments with P. protegens was evident 4 d after application of 1 × 106 cfu/µl. Mortality from B. bassiana was low at 4 d but increased at 5 d. These results imply that P. protegens holds great potential as a biological control agent for incorporation into an integrated pest management program against adult house flies.

Safety of methionine, a novel biopesticide, to adult and larval honey bees (Apis mellifera L.).

Methionine is an essential/indispensible amino acid nutrient required by adult and larval honey bees (Apis mellifera L. [Hymenoptera: Apidae]). Bees are unable to rear broods on pollen deficient in methionine, and reportedly behaviorally avoid collecting pollen or nectar from florets deficient in methioinine. In contrast, it has been demonstrated that methionine is toxic to certain pest insects; thus it has been proposed as an effective biopesticide. As an ecofriendly integrated pest management agent, methionine boasts a novel mode of action differentiating it from conventional pesticides, while providing non-target safety. Pesticides that minimize collateral effects on bees are desirable, given the economic and ecological concerns about honey bee health. The aim of the present study was to assess the potential impact of the biopesticide methionine on non-target adult and larval honey bees. Acute contact adult toxicology bioassays, oral adult assessments and chronic larval toxicity assessments were performed as per U.S. Environmental Protection Agency (EPA) requirements. Our results demonstrated that methionine fits the U.S. EPA category of practically nontoxic (i.e. lethal dose to 50% mortality or LD50 > 11µg/bee) to adult honey bees. The contact LD50 was > 25µg/bee and the oral LD50 was > 100µg/bee. Mortality was observed in larval bees that ingested DL-methionine (effective concentration to 50% mortality or EC50 560µg/bee). Therefore, we conclude that methionine poses little threat to the health of the honey bee, due to unlikely exposure at concentrations shown to elicit toxic effects.

Characterization of a Sodium Channel Mutation in Permethrin-Resistant Rhipicephalus sanguineus (Acari: Ixodidae).

The brown dog tick, Rhipicephalus sanguineus (Latrielle) sensu lato, is an important ectoparasite of dogs and occasionally humans, capable of transmitting several pathogens, such as Rickettsia and Ehrlichia, which are of veterinary and medical importance. The brown dog tick is distributed worldwide and has an affinity for human habitations in much of its range. In some populations, lack of integrated pest management plans and overuse of pyrethroid pesticides and other sodium channel inhibitors has resulted in high levels of resistance to permethrin. Recently, a highly conserved region of the R. sanguineus sodium channel was sequenced, indicating that a single nucleotide polymorphism of thymine to cytosine on domain III segment VI of the sodium channel could confer resistance. A molecular assay targeting a point mutation in the sodium channel was developed and optimized to separate ticks expressing permethrin resistance from those from a susceptible colony. Thereafter, multiple field-collected phenotypically permethrin-resistant populations were evaluated using this molecular assay to determine genotype. As confirmed by DNA sequencing, a point mutation was present at a high rate in phenotypically resistant tick populations that was not present in the susceptible strain. These data suggest an additional permethrin resistance mechanism to metabolic resistance, which has been reported for this tick species, and confirm its association with phenotypic resistance. The results of this study further emphasize the need to preserve acaricide chemistry through rotation of active ingredients used to control ectoparasites.

Laboratory and field evaluation of brown dog tick behavioral responses to potential semiochemicals.

The brown dog tick, Rhipicephalus sanguineus sensu lato (Parasitiformis: Ixodidae), is an ectoparasite of dogs that can be found worldwide. This tick poses unique difficulties in management because it can complete its entire life cycle indoors and has demonstrated acaricide resistance. The ability to monitor for tick presence and abundance is necessary for developing effective control programs. As such, an evaluation of adult brown dog tick behavioral responses to 16 potential semiochemicals was undertaken using Y-tube and straight-tube olfactometers. Both sexes of ticks were activated by nine of the 16 semiochemicals tested, including 300µg of 1-octen-3-ol, benzaldehyde, benzyl alcohol, hexanoic acid, nonanoic acid, methyl salicylate, o-nitrophenol, 2,6-dichlorophenol and salicylaldehyde. Rhipicephalus sanguineus s.l. behaviors including movement speed, direction and duration, as well as turning, were quantified following exposure to the same nine chemicals in a straight-tube olfactometer, individually at 300µg and as mixtures. Three individual chemicals, 1-octen-3-ol, hexanoic acid, and methyl salicylate induced strong responses. These three chemicals were evaluated in a semi-field setting using a modified bed bug trap but were found to provide no significant increase in attraction compared with CO2 alone. The results of these studies provide a foundation for future research regarding semiochemicals of R. sanguineus s.l.

Evaluation of Four Bed Bug Traps for Surveillance of the Brown Dog Tick (Acari: Ixodidae).

The brown dog tick, Rhipicephalus sanguineus (Latrielle), can be a serious residential pest due to its unique ability, among ticks, to complete its lifecycle indoors. A single engorged and fertilized female tick can oviposit around 4,000 eggs, allowing indoor establishment to be rapid and easy to miss in early-stage infestations. Acaricide treatment is currently the primary method of control, but can be costly and can lead to the development of acaricide resistance in tick populations. Traps of various designs can be used to help monitor and manage populations of indoor pests, such as cockroaches and bed bugs, but there are currently no commercially available traps for use with brown dog tick infestations. This study included a comparison of four commercially available bed bug traps (NightWatch [BioSensory Inc., Putnam, CT], Bed Bug Beacon [PackTite, Fort Collins, CO], ClimbUp [Susan McKnight Inc., Memphis, TN], and Verify [FMC Corporation, Philadelphia, PA]) with regard to their efficacy in capturing brown dog ticks, and also compared tick attraction to ClimbUp traps baited with several stimuli including CO2. Significantly more ticks were captured and attracted to the NightWatch and CO2-baited ClimbUp traps than the other two trap models. The results suggest that bed bug traps may be useful in brown dog tick monitoring, and CO2 will likely be an important component of a trapping system employed in the future.

Semiochemicals of the common bed bug, Cimex lectularius L. (Hemiptera: Cimicidae), and their potential for use in monitoring and control.

The recent resurgence of the common bed bug, Cimex lectularius L., has driven an increase in research into the biology and behaviour of this pest. Current control is reliant on the application of insecticides, but, owing to the development of insecticide resistance, there is a need for new tools and techniques. Semiochemicals (behaviour- and physiology-modifying chemicals) could be exploited for management of bed bugs. The aim of this review was to evaluate studies undertaken in bed bug chemical ecology to date, with particular reference to how the research could be exploited for monitoring and control. Bed bugs, like many other insects, have a complex olfactory system. Recent studies have characterised the olfactory sensilla, located on the terminal segment of the antennae, to functional classes by electrophysiological screening. Behavioural studies have revealed the presence of an alarm pheromone and potential airborne aggregation semiochemicals, but it is not yet understood if bed bugs use a sex pheromone during mating. Host location cues have been investigated, and carbon dioxide has been found to be highly attractive both in laboratory and in field studies. Recent field trials have tested blends of other potential kairomones, which have been shown to have an additive effect when used in a heated bed bug trap with carbon dioxide. The trap, which combines heat and kairomones, is the only trap currently available with proven efficacy in the field. In order for semiochemicals to be useful for bed bug management, an increased knowledge and understanding of the biology, behaviour and chemical ecology of this insect is essential.