Insecticide Resistance Development in the Filth Fly Pupal Parasitoid, Spalangia cameroni (Hymenoptera: Pteromalidae), Using Laboratory Selections.

Filth flies remain one of the most prevalent pest groups affecting the animal production industry. Spalangia spp. and Muscidifurax spp. are beneficial parasitic wasps that often are utilized to manage filth fly populations such as house flies, Musca domestica L. (Diptera: Muscidae), and stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae). These wasps search for filth fly pupae as hosts in areas potentially treated with insecticides, which may result in nontarget insecticide selection effects. However, research regarding resistance development in parasitic wasps such as S. cameroni Perkins (Hymenoptera: Pteromalidae) is limited. Therefore, a study was conducted to determine the potential of S. cameroni to develop resistance to the commonly used insecticide permethrin, as well as compare permethrin susceptibility among several S. cameroni strains. After 10 selected generations, susceptibility was significantly lower for the selected strain when compared with that of its unselected parent strain. A comparison of several parasitoid strains collected from different U.S. states indicated that permethrin susceptibility was not significantly different between a baseline strain and more recently established field strains. The potential implications of this previously unrecognized nontarget insecticide exposure effect on filth fly parasitoids are discussed.

Highlights of Veterinary Entomology, 2019.

Research in the field of veterinary entomology is dominated by studies concerning arthropods that affect animal health. In 2019, this research primarily addressed the overwhelming industry need to manage detrimental species such as biting flies and ticks and the ongoing problems caused by insecticide/acaricide resistance evolution in these pests. Research also included evidence supporting the need for the conservation of beneficial species, such as biological control organisms. Many studies in a variety of pest systems have demonstrated the potential detrimental effects of insecticide use on nontarget organisms, and those of veterinary importance are no exception. An emphasis also was placed on research regarding alternative management strategies for veterinary pests. The presentation herein provides a descriptive summary of selected research that contributed greatly to the body of knowledge regarding arthropods of veterinary importance. This included several studies that will pave the way towards more effective veterinary pest management in an effort to improve animal health and welfare and, therefore, the sustainability of animal agriculture.

Behavioral Responses of Cattle to Naturally Occurring Seasonal Populations of Horn Flies (Diptera: Muscidae) Under Rangeland Conditions.

The behavioral responses of cattle under the influence of naturally occurring seasonal horn fly, Haematobia irritans (L.), populations were evaluated under rangeland conditions. This study was replicated four times using 10 cows as the subsampling unit equipped with GPS collars scheduled to receive locational fixes every 5 min for 6 d prior to, and 6 d following horn fly insecticidal control application. Data derived from GPS collars were used to evaluate potential horn fly-induced behavioral modifications expressed during predawn, daytime, and nighttime periods. These data were used to analyze variables, which included distance travelled, daily area explored, vertical and horizontal head movements, and inferred activities such as resting, grazing, and walking. Horn fly populations were estimated using daily visual counts and were reduced significantly on animals following insecticidal application. There was no significant difference between treatment periods in any of the aforementioned analyzed variables. During the night-time hours estimated differences (pretreatment minus posttreatment) for distance travelled, area explored, and vertical head movements were 0.81 ± 0.46 km/d, 0.35 ± 0.21 km(2)/d, and 7.25 ± 5.30 counts/d, respectively. The implications of these observations are discussed.

The ability of selected pupal parasitoids (Hymenoptera: Pteromalidae) to locate stable fly hosts in a soiled equine bedding substrate.

The ability of Spalangia cameroni Perkins, Spalangia endius Walker, and Muscidifurax raptorellus Kogan and Legner to locate and attack stable fly hosts was evaluated under laboratory conditions. Postfeeding third-instar stable fly larvae were released and allowed to pupate in two arena types: large 4.8 liter chambers containing a field-collected, soiled equine bedding substrate; or 120-ml plastic cups containing wood chips. At the time of fly pupariation, parasitoids were released and permitted 72 h to locate and attack hosts. On average, parasitism rates of freely accessible stable fly pupae in cups were not significantly different between parasitoid species. However, parasitism rates in chambers containing either Spalangia spp. were ≈50-fold more than M. raptorellus. Additional intraspecies analysis revealed that parasitism rates both by S. cameroni and S. endius were not significantly different when pupae were freely accessible or within bedding, whereas M. raptorellus attacked significantly more pupae in cups than in the larger chambers where hosts were distributed within bedding. These results suggest that Spalangia spp. are more suited to successfully locate and attack hosts in habitats created by equine husbandry in Florida. Therefore, commercially available parasitoid mixtures containing Muscidifurax spp. may be ineffective if used as a control measure at Florida equine facilities.

Seasonal abundance of stable flies and filth fly pupal parasitoids (Hymenoptera: Pteromalidae) at Florida equine facilities.

Beginning in November 2007 and continuing until December 2009, weekly stable fly, Stomoxys calcitrans (L.), surveillance was conducted at four equine facilities near Ocala, FL, by using alsynite sticky traps for adults and by searching immature developmental sites for pupae. Adult stable fly trap captures were highly variable throughout the year, ranging from 0 to 1,400 flies per trap per farm. The greatest adult stable fly activity was observed during the spring months of March and April, with weekly three-trap means of 121 and 136 flies per farm, respectively. The importance of cultural control measures was most apparent on the only farm with no reported insecticide use and the lowest stable fly trap captures, where an intense daily sanitation and composting program was conducted. A survey of on-site filth fly pupae revealed that 99.9% of all parasitoids recovered were Spalangia spp., consisting of Spalangia cameroni Perkins (56.5%), Spalangia nigroaenea Curtis (34.0%), Spalangia endius Walker (5.8%), and Spalangia nigra Latreille (3.7%). The implications of these findings are discussed.

Identification of a mutation associated with permethrin resistance in the para-type sodium channel of the stable fly (Diptera: Muscidae).

The insect sodium channel is of particular interest for evaluating resistance to pyrethroids because it is the target molecule for this major class of neurotoxic insecticides. The stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), sodium channel coding sequence representing domains IS6 through IVS6 was isolated, and the sequence encoding domain II was compared among individuals of a laboratory strain selected for resistance to permethrin and the unselected, parental generation. A point mutation resulting in a leucine-to-histidine amino acid change was identified (Leul014His), and its location corresponded with that observed for knockdown resistance (kdr) mutations in other insects. As a result, the allele was designated kdr-his. A molecular assay was developed to assess the frequency of this mutation in genomic DNA of individual stable flies from the laboratory selections, which provided further evidence that the kdr-his allele accounts for the observed level ofpermethrin resistance in the selected strain. The assay was then used to evaluate the frequency of the mutation from five field-collected populations originating from three horse farms near Ocala, FL; one horse farm near Gainesville, FL; and one dairy farm near Hague, FL. Frequency of the kdr-his allele ranged from 0.46 to 0.78, supporting further investigation of allele prevalence throughout the stable fly season and in response to field insecticide application.

Host blood meal identification by multiplex polymerase chain reaction for dispersal evidence of stable flies (Diptera:Muscidae) between livestock facilities.

A species-specific multiplex polymerase chain reaction targeting the cytochrome b gene of cattle, horses, humans, and dogs was developed to determine the blood meal sources of stable flies, Stomoxys calcitrans (L.), collected from Florida equine facilities. Of 595 presumptive blood-fed stable flies analyzed, successful host amplification was obtained in 350, for a field host-detection efficiency of 58.8%. The majority of analyzed stable flies had fed on cattle (64.6%), followed by horses (24.3%), humans (9.5%), and dogs (1.6%). A survey of animal-enclosed pastures occurring within 3 km of stable fly collection sites revealed that the nearest cattle were between 0.8 and 1.5 km from the four horse farm sampling sites. Cattle-feeding frequencies were greater on farms where cattle were located at distances of 0.8 km, suggesting that between farm differences in host-feeding frequency is related to the number of and distance from a particular host type. Time course evaluations of previously laboratory-fed stable flies demonstrated that host-detection efficiency with this system was 100, 50, and 0% when flies were evaluated at 16, 24, and 48 h postblood feeding, respectively. The results of this study suggest short-term stable fly dispersal of up to 1.5 km in a 48-h time period. The implications of these findings are discussed.

Assessing permethrin resistance in the stable fly (Diptera: Muscidae) in Florida by using laboratory selections and field evaluations.

Insecticide resistance in the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae),has been demonstrated previously, but mostly with insecticides that are no longer used, such as the organochlorines. Resistance to commonly used pyrethroids has been evaluated twice, but only in the midwestern United States. Stable fly susceptibility to a commonly used pyrethroid, permethrin, was determined in Florida to assess the possibility of resistance development. Diagnostic concentration evaluations of three stable fly field strains demonstrated a maximum of 57 and 21% survival to permethrin residues of 3x and 10x the LC99 of a susceptible strain, respectively. Stable flies from an equine facility with no reported insecticide use demonstrated approximately 20% survival with a 3x diagnostic concentration. Despite a distance of 91-km between field collection sites, survival profiles of field-collected stable fly strains were similar. Although an established stable fly colony collected from a local dairy previously expressed low level resistance to permethrin residues, five generations of laboratory permethrin selection increased resistance 15-fold.

Potential vectors of West Nile virus in a semiarid environment: Doña Ana County, New Mexico.

The potential vectors of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in Doña Ana County, NM, were determined during 2004 and 2005. Trapping was conducted using Centers for Disease Control and Prevention miniature light-traps baited with dry ice, and gravid traps baited with a hay infusion. In addition, sentinel chickens were housed at four of the trapping locations to monitor WNV epizootic transmission. In total, 5,576 pools consisting of 115,797 female mosquitoes were tested for WNV by reverse transcription-polymerase chain reaction, of which 152 from 13 mosquito species representing six genera were positive. Culex tarsalis Coquillett, Culex quinquefasciatus Say, Culex erythrothorax Dyar, Aedes vexans (Meigan), and Psorophora columbiae (Dyar & Knab) accounted for 86% of all detections. Based on the frequency of WNV detection, our data indicate primary and secondary vector roles for Cx. tarsalis and Cx. quinquefasciatus, respectively, with Cx. erythrothorax, Ae. vexans, and Ps. columbiae as occasional vectors of WNV in Dofia Ana County. Other species testing positive for the virus included Aedes aegypti (L.), Anopheles franciscanus McCracken, Culex stigmatosoma Dyar, Culiseta inornata (Williston), Ochlerotatus dorsalis (Meigan), Ochlerotatus sollicitans (Walker), Ochlerotatus trivittatus (Coquillett), and Psorophora signipennis (Coquillett). Although they occurred after initial WNV detections in mosquitoes, in total, 21 seroconversions in sentinel chickens were detected during the study.