Association of Salivary Cholinesterase With Arthropod Vectors of Disease.

Acetylcholinesterase (AChE) was previously reported to be present in saliva of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), with proposed potential functions to 1) reduce acetylcholine toxicity during rapid engorgement, 2) modulate host immune responses, and 3) to influence pathogen transmission and establishment in the host. Potential modulation of host immune responses might include participation in salivary-assisted transmission and establishment of pathogens in the host as has been reported for a number of arthropod vector-borne diseases. If the hypothesis that tick salivary AChE may alter host immune responses is correct, we reasoned that similar cholinesterase activities might be present in saliva of additional arthropod vectors. Here, we report the presence of AChE-like activity in the saliva of southern cattle ticks, Rhipicephalus (Boophilus) microplus; the lone star tick, Amblyomma americanum (Linnaeus); Asian tiger mosquitoes, Aedes albopictus (Skuse); sand flies, Phlebotomus papatasi (Scopoli); and biting midges, Culicoides sonorensis Wirth and Jones. Salivary AChE-like activity was not detected for horn flies Haematobia irritans (L.), stable flies Stomoxys calcitrans (L.), and house flies Musca domestica L. Salivary cholinesterase (ChE) activities of arthropod vectors of disease-causing agents exhibited various Michaelis-Menten KM values that were each lower than the KM value of bovine serum AChE. A lower KM value is indicative of higher affinity for substrate and is consistent with a hypothesized role in localized depletion of host tissue acetylcholine potentially modulating host immune responses at the arthropod bite site that may favor ectoparasite blood-feeding and alter host defensive responses against pathogen transmission and establishment.

Rhipicephalus (Boophilus) microplus (Ixodida: Ixodidae) Larvae Collected From Vegetation in the Coastal Wildlife Corridor of Southern Texas and Research Solutions for Integrated Eradication.

The potential for reinvasion of the United States by cattle fever ticks, Rhipicephalus (Boophilus) annulatus and Rhipicephalus microplus (Canestrini), which remain established in Mexico, threatens the viability of the domestic livestock industry because these ticks vector the causal agents (Babesia bovis and Babesia bigemina) of bovine babesiosis. The Cattle Fever Tick Eradication Program safeguards the health of the national cattle herd preventing the reemergence of bovine babesiosis by keeping the United States free of cattle fever ticks. Here, the collection of free-living southern cattle tick, R. microplus, larvae by sweeping flannel flags over vegetation in the wildlife corridor of Cameron and Willacy Counties, TX, is reported. Finding R. microplus larvae on vegetation complements reports of infestations in wildlife hosts inhabiting the southern Texas coastal plains. Land uses and environmental conditions have changed since cattle fever ticks were eradicated from the United States by 1943. These changes complicate efforts by the Cattle Fever Tick Eradication Program to keep cattle in the United States free of the cattle fever tick disease vectors. Current scientific research on technologies that could be used for area-wide management of fever tick larvae in south Texas and how this could be applied to integrated eradication efforts are discussed.

Metastriate Ixodid Life Stages Protected from Predatory Ants in Texas.

Multiple predatory ant species, including the red imported fire ant, Solenopsis invicta (Buren) (Hymenoptera: Formicidae), have been reported to attack ixodids (Ixodida: Ixodidae), but evidence has largely been circumstantial. When living lone star tick, Amblyomma americanum (L.) (Acarina: Ixodidae), eggs, and unfed and blood-engorged larvae, nymphs, and adults were deployed on bait transects with hot dog slices and dead house flies, Musca domestica L., in West, Central, and South Texas. The various ixodid life stages were not attacked while ants were strongly recruited to the hot dog and M. domestica baits. Similarly, when the same ixodid life stages and other baits were placed adjacent to colonies of two ant species (red harvester ant, Pogonomyrmex barbatus (Smith) and the red imported fire ant, Solenopsis invicta Buren) the ixodids were not preyed upon while hot dog slices and dead M. domestica were immediately attacked. Some ant species dragged blood-engorged adult ixodids and eggs away from the colony entrance, where they were originally placed, and discarded them. Evidence and mechanisms for allomone-based ant deterrence in the genera Amblyomma, Dermacentor, and Rhipicephalus (metastriate ixodids) are discussed. Protection of ixodids from predatory ants helps to explain why metastriate ixodids remain problematic worldwide despite the presence of predaceous ants.

Relationships of Salinity, Relative Humidity, Mud Flat Fiddler Crabs, Ants, and Sea Ox-Eye Daisy With Ixodid Distribution and Egg Survival on the South Texas Coastal Plains.

The South Texas coastal plains are mostly dominated by mesquite-thorn scrub from the Mexican border to cattle ranches extending north from Willacy Co. A wildlife corridor on the plains, composed of natural habitat, supports hosts of many ixodid species. Occasional wind (not lunar) tides and infrequent storm surges inundate coastal plain areas with hypersaline water from the Lower Laguna Madre, creating large areas of saline soil. Laboratory and field experiments and observations were used to identify relationships between salinity and other abiotic and biotic factors that influence ixodid distribution. Exposure of lone star tick, Amblyomma americanum (L.), and Rhipicephalus microplus (Canestrini) eggs to hypersaline water is lethal. Although intermittent hypersaline flooding kills ixodid eggs, saline soil was not particularly toxic. When relative humidity is relatively low, desiccation causes high egg mortality on dry soil, regardless of salinity. Substantial year-round populations of mud flat fiddler crabs, Uca rapax (Smith) (Decopoda: Ocypodidae), occur on saline soil and eliminated ≈80% of A. americanum egg masses overnight. On saline and low-salinity soils predatory formicids, including the red imported fire ant, Solenopsis invicta (Buren), were indifferent to the eggs. Saline soils were dominated by the sea ox-eye daisy, Borrichia frutescens (L.) DC, and its color allowed production of a GIS-based map. At least 24.4% of the wildlife corridor supports heavy stands of B. frutescens which is indicative of high U. rapax populations. Ixodid populations were negligible on heavy B. frutescens stands because of associated salt water toxicity, desiccation, and predation by U. rapax on eggs.

Lethal Effects of Silica Gel-Based CimeXa and Kaolin-Based Surround Dusts Against Ixodid (Acari: Ixodidae) Eggs, Larvae, and Nymphs.

As tick resistance to conventional acaricides becomes more common, alternative control tactics are gaining attention. Insecticidal dusts CimeXa and Surround, based on silica gel and kaolin, respectively, were assessed against Amblyomma americanum (L.) (Ixodida: Ixodidae) eggs, larvae, and nymphs in the laboratory. Coverage by the dry dusts, particularly CimeXa, was strongly lethal to larvae and to a lesser extent to nymphs. Larval mortality was also high when larvae crawled across thin layers of CimeXa and, to a lesser extent, Surround dusts. CimeXa was more lethal to nymphs that crawled across a thin layer than Surround. Larval mortality after crawling on dried aqueous suspensions of the dusts for 30 min and for 48 h caused moderate mortality (<80%) regardless of a 10-fold difference in concentration; nymphal mortality was negligible. In a field experiment, CimeXa dust strongly reduced numbers of Gulf coast tick, Amblyomma maculatum (Koch) (Ixodida: Ixodidae), larvae and nymphs by 24 h. Possible application of CimeXa to control other species of ixodid ticks is discussed as well as advantages and disadvantages of using dusts for tick control under field conditions.

Lethal Effects of the Insect Growth Regulator Cyromazine Against Three Species of Filth Flies, Musca domestica, Stomoxys calcitrans, and Fannia canicularis (Diptera: Muscidae) in Cattle, Swine, and Chicken Manure.

The presence of various species of filth flies is a widespread problem where livestock, including poultry, are maintained and where manure accumulates. The house fly, Musca domestica L.; the stable fly, Stomoxys calcitrans (L.); and the little house fly, Fannia canicularis (L.) (each Diptera: Muscidae), the target pests in our study, can mechanically spread diseases, and S. calcitrans can bite cattle, causing losses in meat and milk production. Chemical control is widely used to suppress filth flies, but resistance to conventional insecticides has become problematic. Hence, an alternative approach, insect growth regulators (IGRs), has been adopted by many livestock producers. We assessed the ability of the IGR cyromazine in granular and granular-based aqueous formulations to suppress the three muscid species from developing in poultry, cattle, and swine manure collected from commercial livestock production facilities. Each of the two formulations provided either strong or complete control of the pests for the 4-wk duration of the study, excluding the granular formulation that provides control of only F. canicularis developing in poultry manure for 2 wk. The two cyromazine-based IGR formulations appear to be effective tools that, if rotated appropriately with other insecticides, can be incorporated into integrated pest management strategies for filth fly suppression.

Seasonal Changes in the Caste Distribution of Foraging Populations of Formosan Subterranean Termite in New Orleans, Louisiana.

This study examined the relationship between temperature, precipitation, soil composition, levels of feeding damage, and the caste distribution (workers, soldiers, nymphs) of the Formosan subterranean termite, Coptotermes formosanus Shiraki, collected in underground monitoring stations over a 12 mo period. Because nymphs are the caste that develops into alates, the seasonal abundance of nymphs was examined over a 5 yr period. Numbers of workers, soldiers, and soldier/worker ratio were significantly affected by month. Recruitment and retention of foraging termites in stations was significantly affected by the level of feeding damage. The number of nymphs collected in monitoring stations was highly variable. In the 12 mo test, there was a significant correlation between numbers of nymphs and level of feeding damage, temperature, precipitation, and soil composition. Over a 5 yr period, significantly more nymphs were collected in 2011 than in 2007 and 2008. Peak nymph collections varied from year to year. Overall, peak nymph collections were more likely to occur in Mar., Sept., and Oct. Increasing our knowledge of the environmental factors that influence recruitment and retention of foraging termites in monitoring stations could influence termite bait placement and improve baiting strategies for termite control. Identifying the key factors that cause aggregations of nymphs in underground stations could increase our ability to predict the intensity and location of alate swarms.

Stable Fly, Stomoxys calcitrans (L.), Dispersal and Governing Factors.

Although the movement of stable fly, Stomoxys calcitrans (L.), has been studied, its extent and significance has been uncertain. On a local scale (<13 km), fly movement occurs between host animals and resting sites to feed and mate, mainly at on-farm locations where herbivorous livestock regularly congregate. Small numbers emigrate from livestock congregation sites in search of other hosts and oviposition substrate, mostly within <1.6 km. Such local movement occurs by flight ~90 cm above ground, or with moving livestock. While stable flies are active year-round in warm latitudes, cold winters in temperate areas result in substantial population and activity declines, limiting movement of any sort to warmer seasons. Long-distance dispersal (>13 km) is mainly wind-driven by weather fronts that carry stable flies from inland farm areas for up to 225 km to beaches of northwestern Florida and Lake Superior. Stable flies can reproduce for a short time each year in washed-up sea grass, but the beaches are not conducive to establishment. Such movement is passive and does not appear to be advantageous to stable fly's survival. On a regional scale, stable flies exhibit little genetic differentiation, and on the global scale, while there might be more than one "lineage", the species is nevertheless considered to be panmictic. Population expansion across much of the globe likely occurred from the late Pleistocene to the early Holocene in association with the spread of domesticated nomad livestock and particularly with more sedentary, penned livestock.

Horn Fly, Haematobia irritans irritans (L.), Overwintering.

The horn fly, Haematobia irritans irritans (L.), is an ectoparasitic blood feeder mainly on cattle. Its cosmopolitan distribution extends from boreal and grassland regions in northern and southern latitudes to the tropics. Stress and blood loss from horn flies can reduce cattle weight gain and milk production. Horn flies show substantial plasticity in their response to winter. Populations in warmer, lower latitudes have been reported to overwinter in a state of dormancy, but most overwinter as active adults in normal or reduced numbers. As latitudes increase, winters are generally colder, and correspondingly, larger percentages of horn fly populations become dormant as pharate adults (a post-pupal, pre-emergent stage) or die. Reports on the effect of elevation on horn fly dormancy at high elevations were contradictory. When it occurs, dormancy takes place beneath cattle dung pats and in the underlying soil. The horn fly's mode of dormancy is commonly called diapause, but the collective research on horn fly diapause (behavioral and biochemical) is not conclusive. Understanding the horn fly's overwintering behaviors can lead to development of pre-dormancy insecticide spray strategies in colder latitudes while other strategies must be determined for warmer regions.

Echinopsacetylenes A and B, new thiophenes from Echinops transiliensis.

Two new polyacetylene thiophenes, echinopsacetylenes A and B (1 and 2), were isolated from the roots of Echinops transiliensis. The structures of 1 and 2 were elucidated on the basis of spectroscopic analyses and chemical transformations. Echinopsacetylenes A (1) is the first natural product possessing an α-terthienyl moiety covalently linked with another thiophene moiety. Echinopsacetylenes B (2) is the first natural thiophene conjugated with a fatty acid moiety. Echinopsacetylene A (1) showed toxicity against the Formosoan subterranean termite (Coptotermes formosanus).

Areawide field study on effect of three chitin synthesis inhibitor baits on populations of Coptotermes formosanus and Reticulitermes flavipes (Isoptera: Rhinotermitidae).

Periodic sampling of 43 independent monitors, initially active with Formosan subterranean termite, Coptotermes formosanus Shiraki, or the eastern subterranean termite, Reticulitermes flavipes (Kollar) (Isoptera: Rhinotermitidae), was conducted to evaluate the effects of cellulose baits containing one of three chitin synthesis inhibitors (CSIs)-diflubenzuron, hexaflumuron, or chlorfluazuron-on termite populations. Diflubenzuron at 0.1% active ingredient (AI, wt:wt) had no noticeable effect on termite populations. Chlorfluazuron (0.25% [AI]) significantly reduced termite populations in approximately 3 yr. Chlorfluazuron used after > 2-yr diflubenzuron treatment significantly reduced termite populations within months. This suggests diflubenzuron exposure increased the termite's sensitivity to chlorfluazuron accelerating population collapse. Hexaflumuron (0.5% [AI]) also reduced termite populations in approximately 2 yr. The process of removing most detectable termite populations from the approximately 160,000-m2 campus of the Southern Regional Research Center, New Orleans, LA, with CSIs baits required approximately 3 yr. Adjustments in the specific bait formulations and application procedures might reduce time to suppression. Establishment of new independent termite populations provides a mechanism to minimize the effects of baits. Remedial control measures around and under structures should be considered when implementing an area wide management strategy.

Effect of seasonal changes in soil temperature and moisture on wood consumption and foraging activity of Formosan subterranean termite (Isoptera: Rhinotermitidae).

The objective of this study was to determine how seasonal changes affect the foraging activity and wood consumption of the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), in New Orleans, LA. There was a significant correlation between wood consumption and air temperature, soil temperature, and soil moisture, but not precipitation or number of rainy days. In the first year of the study, wood consumption was the lowest in December, February, and March. Wood consumption in January was not significantly different from consumption during the rest of the year. There were no seasonal changes in the number of underground monitoring stations occupied by termites. In the second year of the study, wood consumption was lowest from January to March. There was a significant decrease in the number of monitoring stations occupied by termites during the winter. This study determined that C. formosanus will remain in monitoring stations and resume feeding during warmer periods of a mild winter if average soil temperatures remain above 15 degrees C. Only prolonged periods of cold weather, with average soil temperatures below 15 degrees C, caused a significant number of termites to abandon underground monitoring stations. Seasonal changes in foraging activity would probably only disrupt baiting programs during severe winters in New Orleans, LA.

Influence of dry soil on the ability of Formosan subterranean termites, Coptotermes formosanus, to locate food sources.

The effect of barriers of dry soil on the ability of Formosan subterranean termites, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), to construct tunnels and find food was evaluated. Termite movement and wood consumption in a three-chambered apparatus were compared between treatments with dry soil in the center container and treatments where the soil in the center container was moist. When a wood block was located in the release container, termites fed significantly more on that block, regardless of treatment or soil type. In the treatment with dry clay, none of the termites tunneled through the dry clay barrier to reach the distal container. When termites had to tunnel through a barrier of dry sand, topsoil, or clay to reach the sole wood block, there was no effect on wood consumption for the sand treatment, but there was significantly less feeding on wood in the treatments with dry topsoil or clay. When foraging arenas had a section of dry sand in the center, the dry sand significantly reduced tunneling in the distal section after 3 days, but not after 10 days. There was a highly significant effect on the ability of termites to colonize food located in dry sand. Only one feeding station located in dry sand was colonized by termites, compared with 11 feeding stations located in moist sand.

Effect of soil type and moisture availability on the foraging behavior of the Formosan subterranean termite (Isoptera: Rhinotermitidae).

This study examined the influence of soil type and moisture availability on termite foraging behavior. Physical properties of the soil affected both tunneling behavior and shelter tube construction. Termites tunneled through sand faster than top soil and clay. In containers with top soil and clay, termites built shelter tubes on the sides of the containers. In containers with sand, termites built shelter tubes directly into the air and covered the sides of the container with a layer of sand. The interaction of soil type and moisture availability affected termite movement, feeding, and survival. In assays with moist soils, termites were more likely to aggregate in top soil over potting soil and peat moss. However, termites were more likely to move into containers with dry peat moss and potting soil than containers with dry sand and clay. Termites were also significantly more likely to move into containers with dry potting soil than dry top soil. In the assay with dry soils, termite mortality was high even though termites were able to travel freely between moist sand and dry soil, possibly due to desiccation caused by contact with dry soil. Evaporation from potting soil and peat moss resulted in significant mortality, whereas termites were able to retain enough moisture in top soil, sand, and clay to survive for 25 d. The interaction of soil type and moisture availability influences the distribution of foraging termites in microhabitats.

Effects of flooding on field populations of Formosan subterranean termites (Isoptera: Rhinotermitidae) in New Orleans, Louisiana.

Hurricane Katrina (2005) resulted in extensive flooding in the city of New Orleans, LA. Periodic sampling of monitors before the flood, and of different monitors in the same areas after the flood, was used to evaluate the effects of long-term flooding on populations of Formosan subterranean termites, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Monitors were located adjacent to buildings and in urban forests. Significant population reductions occurred in areas that flooded 2-3 wk with brackish water, with termite populations associated with pine (Pinus spp.) trees and buildings slower to recover than populations associated with oak trees. Alate production in flooded areas showed no reduction from previous years.

Activity of 1,4-benzoquinones against formosan subterranean termites (Coptotermes formosanus).

A large number of naturally occurring and synthetic benzoquinones were evaluated for activity against the Formosan subterranean termite, Coptotermes formosanus, with potential use in termite control. Among these bioactive naturally occurring benzoquinones are 2-methyl-5-isopropyl-1,4-benzoquinone, 2-methoxy-6-pentyl-1,4-benzoquinone, 2,3-dimethoxy-5-methyl-6-(3-methyl-2-butenyl)-1,4-benzoquinone, 2,3-dimethoxy-5,6-dimethyl-1,4-benzoquinone, and 2,3-dichloro-5,6-dimethyl-1,4-benzoquinone. All five of these compounds demonstrated 100% mortality against C. formosanus by day 11 at a concentration of 1% (wt/wt) or less. In general, benzoquinones with one or two hydrophobic groups on the 5 and/or 6 positions of the quinone ring along with one or two group(s) on the opposite side of the ring, at the 2 and/or 3 position, led to high rates of mortality against C. formosanus. Quantitative structure-activity relationship (QSAR) studies showed no correlation between lipophilicity (calculated log P) and mortality for the entire group of nonhalogenated benzoquinones. A correlation was observed between C-6 chain length and day 3 percent mortality for 2,3-dimethoxy-5-methyl-6-substituted aliphatic benzoquinones where short chain lengths resulted in higher mortality.

Phytotoxic Eremophilanes from Ligularia macrophylla.

Systematic bioassay-guided fractionation of the methylene chloride extract of the roots from Ligularia macrophylla was performed to identify both phytotoxic and antifungal compounds. Four phytotoxic eremophilanes (furanoeremophilan-14beta,6alpha-olide, 6beta-angeloyloxy-10beta-hydroxyfuranoeremophilane, eremophil-7(11)-ene-12,8alpha;14beta,6alpha-diolide, and 3alpha-angeloyloxybakkenolide A) and two antifungal fatty acids (linoleic acid and alpha-linolenic acid) were isolated. The X-ray crystal structure determination of 6beta-angeloyloxy-10beta-hydroxyfuranoeremophilane is reported here for the first time. All four eremophilanes substantially inhibited growth of the monocot Agrostis stolonifera (bentgrass) while demonstrating little activity against the dicot Lactuca sativa (lettuce) at 1000 microM. In a dose-response screening of all compounds for growth inhibitory activity against Lemna paucicostata, 6beta-angeloyloxy-10beta-hydroxyfuranoeremophilane was the most active with an IC50 of 2.94+/-0.16 microM. This compound also caused the greatest reduction of photosynthetic electron flow; however, its mode of action remains to be determined. Evaluation of isolated compounds for activity against the Formosan subterranean termite, Coptotermes formosanus, is also reported. At a concentration of 0.5% (wt/wt), 6beta-angeloyloxy-10beta-hydroxyfuranoeremophilane significantly reduced the consumption of filter paper by C. formosanus.

Antifeedant and toxicity effects of thiophenes from four Echinops species against the Formosan subterranean termite, Coptotermes formosanus.

Over 220 crude extracts from repositories generated from plants native to Greece and Kazakhstan were evaluated for termiticidal activity against the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Emerging from this screening effort were bioactive extracts from two Greek species (Echinops ritro L. and Echinops spinosissimus Turra subsp. spinosissimus) and extracts from two Kazakhstan species (Echinops albicaulis Kar. & Kir. and Echinops transiliensis Golosh.). Fractionation and isolation of constituents from the most active extracts from each of the four species has been completed, resulting in the isolation of eight thiophenes possessing varying degrees of termiticidal activity. 2,2':5',2"-Terthiophene and 5'-(3-buten-1-ynyl)-2,2'-bithiophene demonstrated 100% mortality against C. formosanus within 9 days at 1 and 2 wt% concentrations respectively. In addition, all but two of the eight compounds tested were significantly different from the solvent controls in the filter paper consumption bioassay.

Effect of imidacloprid soil treatments on occurrence of Formosan subterranean termites (Isoptera: Rhinotermitidae) in independent monitors.

Periodic sampling of 30 independent monitors, initially active with the Formosan subterranean termite, Coptotermes formosanus Shiraki, was conducted to evaluate the effects of soil treated with imidacloprid on nearby termite activity. Monitors were located adjacent (1-3 m) to the buildings. Soil around and under the buildings was treated with 0.05% imidacloprid. None of the termites collected showed latent mortality attributed to imidacloprid intoxication. Imidacloprid soil treatments did not measurably reduce C. formosanus populations adjacent to the treatments. Imidacloprid does not seem to fit the liquid-bait model.

United States Department of Agriculture-Agriculture Research Service research on targeted management of the Formosan subterranean termite Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae).

The Formosan subterranean termite, Coptotermes formosanus Shiraki is currently one of the most destructive pests in the USA. It is estimated to cost consumers over US dollars 1 billion annually for preventative and remedial treatment and to repair damage caused by this insect. The mission of the Formosan Subterranean Termite Research Unit of the Agricultural Research Service is to demonstrate the most effective existing termite management technologies, integrate them into effective management systems, and provide fundamental problem-solving research for long-term, safe, effective and environmentally friendly new technologies. This article describes the epidemiology of the pest and highlights the research accomplished by the Agricultural Research Service on area-wide management of the termite and fundamental research on its biology that might provide the basis for future management technologies. Fundamental areas that are receiving attention are termite detection, termite colony development, nutrition and foraging, and the search for biological control agents. Other fertile areas include understanding termite symbionts that may provide an additional target for control. Area-wide management of the termite by using population suppression rather than protection of individual structures has been successful; however, much remains to be done to provide long-term sustainable population control. An educational component of the program has provided reliable information to homeowners and pest-control operators that should help slow the spread of this organism and allow rapid intervention in those areas which it infests.