A new formulation of screwworm fly attractant with reduced hazardous chemicals and transport restrictions.

New World screwworm flies, Cochliomyia hominivorax (Coquerel), are obligate parasites of warm-blooded animals. They were eradicated from North and Central America during the mid-20th to early-21st centuries using the sterile insect technique (SIT), a method presently employed to maintain a permanent barrier between Central and South America. Lures are an important component of the screwworm eradication program, where they are used for surveillance, sample collection, and strain evaluation in the field. The first chemical lure, later named swormlure, was developed based on the attractiveness of C. hominivorax to volatile organic compounds (VOCs) produced from decomposing animal tissues. The formulation has changed little over the years and presently contains 10 chemicals, one of which is dimethyl disulfide (DMDS). Restrictions on the transport of DMDS have recently impeded its use in swormlure-4 (SL-4). However, dimethyl trisulfide (DMTS) is not as highly restricted and can be shipped via air transportation. Both chemicals are produced by microbial decomposition of animal tissues. Here, we conducted field trials using three releases of sterile C. hominivorax, each comprising approximately 93,000 flies, to test the efficacy of SL-4, containing DMDS, to swormlure-5 (SL-5) containing DMTS. Traps baited with SL-4 and SL-5 captured 575 (mean = 191.7, SD 17.9) and 665 (mean = 221.7, SD 33.2) C. hominivorax, respectively (df = 19, F = 1.294, P = 0.269). However, traps baited with SL-5 captured considerably more Cochliomyia macellaria (Fabricius), a closely related but nontarget fly.

Geographic Population Genetic Structure of the New World Screwworm, Cochliomyia hominivorax (Diptera: Calliphoridae), Using SNPs.

The New World screwworm, Cochliomyia hominivorax (Coquerel 1858) (Diptera: Calliphoridae), is a serious parasite of livestock, humans, and other warm-blooded animals. It has been eradicated from the northern parts of its historical range down to the Panama-Colombian border where a permanent barrier zone is maintained. This eradication was accomplished through using the sterile insect technique (SIT). In 2016 there was an outbreak of C. hominivorax in the Florida Keys. In only six months, this pest was successfully re-eradicated using SIT, but the geographic origin of the invasion has yet to be resolved. It was previously determined that the Florida flies most likely represented a single invasion, and it was recommended that a finer-scale genetic assessment should be completed. Thus, this current proof-of-concept study aimed to develop a population genetic database using single nucleotide polymorphisms (SNPs) to reference outbreaks and potentially identify the origin of the Florida outbreak. This initial database consists of wild-caught samples from 4 geographic locations as well as laboratory colony samples that originated from 7 additional locations using a genotyping by sequencing (GBS) approach. Geographic population structuring was identified for twelve populations that clustered according to geographic location. The Florida outbreak samples appeared similar to samples from the outer Caribbean cluster which included samples from Dominican Republic and Trinidad and Tobago, however, these results will be further clarified with the replacement of laboratory colony samples with future wild-caught samples.

The microbiome of wild and mass-reared new world screwworm, Cochliomyia hominivorax.

Insect population control through continual releases of large numbers of sterile insects, called sterile insect technique (SIT), is only possible if one can mass-rear large quantities of healthy insects. Adaptation of insect stocks to rearing conditions and artificial feeding systems can have a multitude of negative effects such as inbreeding depression, reduced compatibility with wild strains, unintentional selection for traits that lower fitness after release, and an altered microbiome. Changes to insect microbiomes can have many effects on insects ranging from a reduction in sex pheromones or reduced fitness. Thus understanding these systems is important for mass rearing and the performance of the sterile insect control programs. In this study we explored the microbiome of the New World screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae) an economically important parasite of warm-blooded animals. Samples from myiases in cows and wild adults were compared to and mass-reared flies used by the SIT program. Significant differences were observed between these treatments, with wild captured flies having a significantly more diverse microbial composition. Bacteria known to stimulate oviposition were found in both wild and mass-reared flies. Two bacteria of veterinary importance were abundant in wild flies, suggesting screwworm is a potential vector of these diseases. Overall, this study provides the screwworm eradication program a platform to continue exploring the effects associated bacteria have on screwworm fitness.

Use of a multiagency approach to eradicate New World screwworm flies from Big Pine Key, Florida, following an outbreak of screwworm infestation (September 2016-March 2017).

On September 30, 2016, the US National Veterinary Services Laboratory confirmed an autochthonous case of New World screwworm infestation in a Key deer (Odocoileus virginianus clavium) from Big Pine Key, Fla. This case marked the first identification of a sustained and reproducing population of New World screwworm flies in the United States since 1966. Multiple federal, state, and local government agencies collaborated to initiate a response to the outbreak. Efforts were successful in eradicating the flies from Florida.

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.

Screwworm (Diptera: Calliphoridae) in the United States: Response to and Elimination of the 2016-2017 Outbreak in Florida.

Eradicating screwworm, Cochliomyia hominivorax (Coquerel), from continental North American via the sterile insect technique has provided huge economic benefit to livestock producers by eliminating screwworm myiasis. After confirmatory identification of fly samples from infested deer by the USDA National Veterinary Services Laboratory on September 30, 2016, an alert was issued that screwworm myiasis was discovered in the Florida Keys. Personnel from USDA Animal and Plant Health Inspection Service, Agricultural Research Service, the State of Florida, U.S. Fish and Wildlife Service and local officials responded to the outbreak focus on Big Pine Key. After witnessing infested Key deer (Odocoileus virginianus clavium Barboyr & Allen), screwworm adult sampling was initiated at 0930 h on October 5, 2016 using nets to collect flies arriving at putrid liver, with the first female collected within 1 h. Larval samples were collected from infested animals for DNA analyses and to develop a "Florida outbreak" colony to test mating compatibility with the mass-produced strain used for sterile fly releases. Ground release chambers for sterile screwworm releases were placed in favorable habitats based on satellite image analyses. Sterile pupae were first placed in the chambers on October 11, 2016. Further liver trapping showed that 13 Keys were infested. One case, presumably through animal movement, occurred near Homestead on the Florida mainland. Ultimately there were 35 sterile fly release stations, including 4 located around Homestead, but no further cases were identified. About 188 million sterile flies were released until successful eradication was declared on March 23, 2017. Containing the outbreak prevented economic losses to livestock producers and other wildlife on the mainland and kept eradication costs to a minimum.

Molecular Characterization of the 2016 New World Screwworm (Diptera: Calliphoridae) Outbreak in the Florida Keys.

New World screwworm (NWS), Cochliomyia hominivorax (Coquerel 1858) (Diptera: Calliphoridae), is a myiasis-causing fly that can be a serious threat to the health of livestock, wildlife, and humans. Its progressive eradication from the southern United States, Mexico, and Central America from the 1950s to 2000s is an excellent example of successful pest management using sterile insect technique (SIT). In late 2016, autochthonous NWS were detected in the Florida Keys, representing this species' first invasion in the United States in >30 yr. Rapid use of quarantine and SIT was successful in eliminating the infestation by early 2017; however, the geographic source of this infestation remains unknown. Here, we use amplicon sequencing to generate mitochondrial and nuclear sequence data representing all confirmed cases of NWS from this infestation, and compare these sequences to preexisting data sets sampling the native distribution of NWS. We ask two questions regarding the FL Keys outbreak. First, is this infestation the result of a single invasion from one source, or multiple invasions from different sources? And second, what is the geographic origin of this invasion? We found virtually no sequence variation between specimens collected from the FL Keys outbreak, which is consistent with a single source of introduction. However, we also found very little geographic resolution in any of the data sets, which precludes identification of the source of this outbreak. Our lack of success in answering our second question speaks to the need for finer-scale genetic or genomic assessments of NWS population structure, which would facilitate source determination of potential future outbreaks.

Distribution and Persistence of Sterile Screwworms (Diptera: Calliphoridae) Released at the Panama-Colombia Border.

The sterile insect technique is used by the Comisión Panamá - Estados Unidos para la Erradicación y Prevención del Gusano Barrenador del Ganado (COPEG) to maintain a barrier at the border of Panama and Colombia to prevent screwworms, Cochliomyia hominivorax (Coquerel), from South America reinfesting North America. Before studying the distribution and persistence of sterilized, mass-produced screwworms released in the barrier zone, the utility of applying fluorescent dust (∼1.0 mg/fly) to pupae and to newly emerged adults was evaluated to determine the potential effect on fly survival. The flight ability of flies collected from two adult emergence/collection systems (enclosed towers and open chambers) and treated with low (∼0.20 mg/fly) or high (∼1.0 mg/fly) amounts of fluorescent powder was compared. The distribution and persistence of sterile screwworms marked with fluorescent powder (∼0.20 mg/fly), after collection from the same two adult emergence/collection systems, was compared after their release in the barrier zone. The results demonstrated that: 1) fluorescent dust did not negatively affect sterile screwworm longevity or flight ability; 2) no differences were detected between sterile flies collected from the two emergence systems; and 3) sterile screwworms distributed evenly in the barrier zone and persisted for > 6 d. This information was useful in implementing the use of a new sterile fly emergence/collection system and deploying a new strain by COPEG for the barrier zone maintenance program; it will be valuable for evaluating alternative release strategies of sterile screwworms by the eradication and barrier maintenance program.

A transgenic male-only strain of the New World screwworm for an improved control program using the sterile insect technique.

BACKGROUND: The New World screwworm, Cochliomyia hominivorax, is a devastating pest of livestock endemic to subtropical and tropical regions of the Western hemisphere. The larvae of this species feed on the tissue of living animals, including man, and can cause death if untreated. Over 60 years ago, the sterile insect technique (SIT) was developed with the aim of eradicating this pest, initially from Florida but subsequently from all of North and Central America. From the outset it was appreciated that SIT would be more efficient if only sterile males were released in the field, but this was not possible until now. RESULTS: Here, we report on the development and evaluation of the first sexing strains of C. hominivorax that produce only males when raised on diet without tetracycline. Transgenic lines have been developed that possess a tetracycline repressible female-lethal genetic system. Ten of these lines show high female lethality at the late larval/pupal stages and three of them present dominant female lethality. Most of the lines were comparable to the wild type parental strain in several fitness parameters that are relevant to mass rearing in a production facility. Further, three lines performed well in male mating success and male competition assays, suggesting they would be sexually competitive in the field. Consequently, one transgenic line has been selected by the New World Screwworm Program for evaluation under mass rearing conditions. CONCLUSIONS: We conclude that the promising characteristics of the selected sexing strains may contribute to reduce production costs for the existing eradication program and provide more efficient population suppression, which should make a genetic control program more economical in regions were C. hominivorax remains endemic.

Development of a spatially targeted field sampling technique for the southern cattle tick, Rhipicephalus microplus, by mapping white-tailed deer, Odocoileus virginianus, habitat in South Texas.

The objective of our study was to determine whether satellite remote sensed data could be used to identify white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), habitat and target locations for sampling free-living larvae of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae) in South Texas. Two methods for mapping white-tailed deer habitat were used, an object-oriented method to identify closed canopies and waterways for deer movement and two vegetation indices: the Normalized Difference Vegetation Index and the Modified Soil Adjusted Vegetation Index to identify forage for deer. These two data sets of favorable white-tailed deer habitat were combined within a geographic information system to identify locations for sampling ticks. Larvae of R. (B.) microplus, were sampled in Zapata County, Texas, by walking transects with attached flannel panels to jeans. Although the data set and sampling period were limited, data analysis demonstrated that sampling of free-living larvae of R. (B.) microplus can be conducted in South Texas, and larvae were most abundant in areas that harbored O. virginianus. Spatial analysis of satellite imagery to classify white-tailed deer/southern cattle tick habitat proved efficacious and may be useful in directing sampling activities in the field.

Handling stairs in the seated position for people with unilateral lower-limb amputations.

Handling stairs in the seated position for people with unilateral lower-limb amputations. The objective of this uncontrolled pilot study was to test the hypotheses that the seated stair-handling method enables people with unilateral lower-limb amputations to ascend and descend stairs effectively and safely, and with an acceptable level of perceived exertion. Eight people with unilateral lower-limb amputations each received a single 20-minute education session on climbing stairs in the seated position. The main outcome measures, assessed 3 days after training, were success in ascending and descending a flight of 11 stairs, safety, and rating of perceived exertion (Borg CR-10 scale, range 0-10). Before training, 3 participants (37.5%) were successful in using a variety of methods. After training, all 8 (100%) were successful, but only 7 (87.5%) were successful when required to use the seated stair-handling method. Two participants had difficulty with this method, one as a result of arm weakness and the other as a result of aggravation of knee pain. There were no adverse events. Ratings of perceived exertion for the seated method (n=7) ranged from 3 to 7.5 for stair ascent and 1 to 5.5 for descent. The seated stair-handling method is a generally effective, safe, and well-tolerated method for people with unilateral lower-limb amputations to ascend and descend stairs.

Seasonal and spatial distributions of adult screwworms (Diptera: Calliphoridae) in the Panama Canal area, Republic of Panama.

The distribution of screwworms, Cochliomyia hominivorax, (Coquerel) was studied in a seasonally moist lowland tropical forest in the Republic of Panama using a combination of field collections and satellite imagery. We found that different forest types could be distinguished and mapped using remotely sensed data. To determine the temporal and spatial distribution of flies, we collected flies coming to rotted liver at 82 sites in ten vegetation types (open areas, edge forest, dry scrub forest, forest successional stage 1, forest successional stage 2, forest successional stage 3, forest successional stage 4, forest successional stage 5, mature forests, palm swamp forest, and forest along streams) over three seasons (dry, transitional, wet). Nine of the vegetation types (excluding dry scrub forest) were identified and mapped using SPOT XS and Landsat 5 TM satellite data. Screwworm flies were most abundant during the transition from wet to dry season. Fly numbers were consistently higher in forest habitats, particularly those with trees 20-30 m in height and a fairly open canopy composed of many deciduous species that shed their leaves during the dry season. Screwworm numbers were also high in palm swamp forest, edge forest, and mature growth forest. Traps sampled in open areas had fewer flies and were unrelated to proximity to cattle. Females accounted for 88% of the total fly counts. This study further substantiates the importance of forests in the ecology and behavior of screwworm flies and demonstrates that remotely sensed data can be used to construct the spatial distribution of these flies in a tropical landscape. We discuss implications of this information to the screwworm eradication program.