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Culicoides sonorensis (Wirth & Jones) is the principal North American vector of bluetongue virus (BTV). BTV infection of livestock is distinctly seasonal (late summer and fall) in temperate regions of the world such as California, which has led to speculation regarding vertical transmission of the virus within the midge vector as a potential mechanism for interseasonal maintenance ("overwintering") of the virus. To evaluate potential vertical transmission of BTV in its midge vector, we fed adult midges BTV-spiked blood and used a BTV-specific quantitative reverse transcriptase polymerase chain reaction assay to evaluate parent, egg, and progeny stages of laboratory-reared C. sonorensis for the presence of viral nucleic acid. Whereas BTV nucleic acid was weakly detected in egg batches of virus-fed female midges, virus was never detected in subsequent progeny stages (larvae, pupae, and F1 generation adults). Similarly, BTV was not detected in pools of larvae collected from the waste-water lagoon of a BTV-endemic dairy farm in northern California during the seasonal period of virus transmission. Collectively, these results indicate that BTV is not readily transmitted vertically in C. sonorensis, and that persistence of the virus in long-lived parous female midges is a more likely mechanism for overwintering of BTV in temperate regions.

Heleidomermis magnapapula parasitizes the blood-feeding midge Culicoides sonorensis . Most (84%) single mermithid infective second stage juveniles (J2) developed into adult females, while parasitism by multiple J2 yielded 97% male adults. Nematodes emerged from the midge larval host as adults and mated immediately; females were ovoviviparous. Host larvae were exposed to nematode J2 and examined intact microscopically to score initial parasite load. Midge hosts were reared individually. Premature midge death, nematode survival within the host, and emerging adult nematode sex ratio and size as a function of load and host size were all tracked. Higher nematode loads produced smaller adult nematode males. The higher loads also increased and accelerated premature host death. Emergence of > 7-9 adult nematode males was rare, but up to 19 tiny males emerged from a single host. Larger midges supported higher parasite loads and a larger total volume of emerged nematode biomass. Virgin adult nematode males then were paired with females of variable, known sizes (volume) and held to determine size effects on fertility (egg hatch), and male survival (longevity). Tested adult males ranged in size from 0.0025 - 0.0334 mm 3 and females from 0.0121 - 0.1110 mm 3 . Logistic regression indicated female nematode fertility was positively influenced by male nematode size, while nematode load and female nematode size had no significant effect. While fertility was reduced statistically in smaller males, even some of the smallest male and female individuals could be fertile. Findings are related to field studies in this system., (© 2023 Bradley A. Mullens et al., published by Sciendo.)

Tabanus variegatus F. 1805 has been called by the name Tabanus sulcifronsMacquart 1855 for over 80 yr; T. variegatus is one of the most common large horse flies attacking livestock in much of the southeastern U.S. Morphological, ecological, and molecular evidence indicates that T. variegatus is a distinct species, and we redescribe the female and describe the male. The Fabricius holotype, heavily damaged after over nearly 220 yr, is nevertheless taxonomically sound. Morphology (size, color, palp shape, and r5 wing cell shape) can usually distinguish T. variegatus from T. sulcifrons, but some specimens remain difficult to separate, especially in and west of the Mississippi River Valley. Using geometric morphometric analyses of the wing vein arrangement and palp shape the two species are significantly different. The wings of T. variegatus females also have more microsetae and sometimes a "frosty" appearance. Where they are common and sympatric, as in eastern Tennessee, they are temporally separated such that T. variegatus flies later (August-October) than T. sulcifrons (June-August), minimizing opportunity for gene flow. Museum specimens allow the approximate range of T. variegatus to be compared with that of T. sulcifrons s.l.; T. variegatus is particularly abundant from the coast of the Carolinas and Georgia east to central Tennessee and south to about central Alabama. DNA evidence (COI gene) recovers T. variegatus and T. sulcifrons s.s. in separate clades. Further studies on the T. sulcifrons complex are needed to fully resolve the range of both species, assess the degree of genetic substructuring, and examine relationships with other members of the T. sulcifrons complex., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The availability of rapid, highly sensitive and specific molecular and serologic diagnostic assays, such as competitive enzyme-linked immunosorbent assay (cELISA), has expedited the diagnosis of emerging transboundary animal diseases, including bluetongue (BT) and African horse sickness (AHS), and facilitated more thorough characterisation of their epidemiology. The development of assays based on real-time, reverse-transcription polymerase chain reaction (RT-PCR) to detect and identify the numerous serotypes of BT virus (BTV) and AHS virus (AHSV) has aided in-depth studies of the epidemiology of BTV infection in California and AHSV infection in South Africa. The subsequent evaluation of pan-serotype, real-time, RT-PCR-positive samples through the use of serotype-specific RT-PCR assays allows the rapid identification of virus serotypes, reducing the need for expensive and time-consuming conventional methods, such as virus isolation and serotype-specific virus neutralisation assays. These molecular assays and cELISA platforms provide tools that have enhanced epidemiologic surveillance strategies and improved our understanding of potentially altered Culicoides midge behaviour when infected with BTV. They have also supported the detection of subclinical AHSV infection of vaccinated horses in South Africa. Moreover, in conjunction with whole genome sequence analysis, these tests have clarified that the mechanism behind recent outbreaks of AHS in the AHS-controlled area of South Africa was the result of the reversion to virulence and/or genome reassortment of live attenuated vaccine viruses. This review focuses on the use of contemporary molecular diagnostic assays in the context of recent epidemiologic studies and explores their advantages over historic virus isolation and serologic techniques.

The two most economically important poultry ectoparasites are the northern fowl mite, Ornithonyssus sylviarum (Canestrini and Fanzago), and the poultry red mite, Dermanyssus gallinae (De Geer). Both mites are obligate blood feeders but differ in where they reside. Sampling methods thus focus on-host, especially the vent feathers, for northern fowl mite and off-host, especially cracks and crevices near the nighttime roosting areas, for poultry red mite. Much remains unknown, however, about the basic biology and ecology of both mites. Here we discuss mite detection, quantification, and decision making and provide thoughts on future directions for research., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.)

The northern fowl mite (NFM) Ornithonyssus sylviarum Canestrini and Fanzago is a blood-feeding ectoparasite found on many wild bird species and is a pest of poultry in the United States. It is unknown where NFM infestations of poultry originate, which has made it difficult to establish preventative biosecurity or effective control. We used microsatellite markers to evaluate genetic variation within and among NFM populations to determine routes of introduction onto farms and long-term persistence. We compared NFM from flocks of chickens (Gallus gallus) on different farms in California, Washington, and Georgia, and we compared NFM collected over a 5-yr interval. On three farms we collected NFM from chickens and house sparrows (Passer domesticus) nesting on each farm, which we used to assess movement between host species. There was strong genetic structure among mites from different poultry farms and low estimates of migration between farms. There were significant differences between mites on chickens and house sparrows on two farms where sparrows nested near flocks, indicating no exchange of mites. Only one farm showed evidence of NFM movement between chickens and sparrows. There was high genetic similarity between mites collected 5 yr apart on each of two farms, indicating that NFM infestations can persist for long periods. The genetic patterns did not reveal sources of NFM infestations on chicken farms. The data suggest that NFMs are strongly differentiated, which likely reflects periodic population declines with flock turnover and pesticide pressure., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Stable isotope labeling is a promising method for use in insect mark-capture and dispersal studies. Culicoides biting midges, which transmit several important animal pathogens, including bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV), are small flies that develop in various semi-aquatic habitats. Previous Culicoides dispersal studies have suffered from the limitations of other labeling techniques, and an inability to definitively connect collected adult midges to specific immature development sites., Results: Adult C. sonorensis were successfully labeled with 13 C and 15 N stable isotopes as larvae developing in a semi-aquatic mud substrate in the laboratory. High and low-dose isotope treatments for both elements significantly enriched midges above the background isotope levels of unenriched controls. Enrichment had no effect on C. sonorensis survival, though a slight (~ 5 day) delay in emergence was observed, and there was no significant effect of pool size on 13 C or 15 N enrichment levels., Conclusions: Stable isotope labeling is life-long, and does not interfere with natural insect behaviors. Stable isotope enrichment using 13 C or 15 N shows promise for Culicoides dispersal studies in the field. This method can be used to identify adult dispersal from larval source habitat where a midge developed. It may be possible to detect a single enriched midge in a pool of unenriched individuals, though further testing is needed to confirm the sensitivity of this method.

Haematobia irritans (L.) (Diptera: Muscidae) were netted from pastured cattle in California over two summers, with the goal of documenting on-host sex ratios and mating success as they might vary with repellent applications. Sex ratios were calculated from 100 adults per herd per date, and mating status of females (n = 30 per herd per date) was assessed by dissection of spermathecae. Flies were collected before, during, and after 2-wk treatment periods with each of two repellent-oil mixtures: a 15% mixture of fatty acids (C8, C9, C10) in mineral oil or a 2% solution of geraniol in mineral oil. During the treatment period, flies were collected 3-4 d after the prior treatment and immediately before the next treatment. A separate herd used in the second year was untreated. Overall, the proportion female was 0.65 ± 0.01 (mean ± SE) (range 0.28-0.81). Sex ratios were not altered over time by treatments with either repellent. Overall, proportion of females mated was 0.744 ± 0.018 (range 0.40-1.00). Mating status was significantly affected by the two repellent treatments, which did not differ from each other. For treatment herds, proportion mated before treatment was 0.789 ± 0.039, dropping significantly to 0.659 ± 0.038 during the treatment period, and rebounding to 0.750 ± 0.035 after treatments ceased. Treatments probably killed or repelled the existing heterogeneous fly population, and during treatment relatively more flies were unmated recent arrivals.

The egg and larval instars (L1-L4) of Culicoides sonorensis Wirth and Jones are described. Average (range) of body lengths of a laboratory colony (Van Ryn) were 0.45 (0.43-0.48) mm for eggs, 0.72 (0.39-0.92) mm for L1, 1.8 (1.3-2.2) mm for L2, 3.6 (2.2-5.0) mm for L3, and 5.9 (5.0-6.9) mm for L4. Head capsule lengths were 0.067 (0.064-0.070) mm for L1, 0.100 (0.094-0.110) for L2, 0.157 (0.144-0.168) for L3, and 0.316 (0.300-0.324) mm for L4. Wild and colony L4 larvae differed in many standard metrics such as head length or width, but head ratios and pharyngeal armature measurements were comparable. Standard external and internal features of the head capsule are described. Scanning electron microscope photos revealed pharyngeal armature structure was similar for L2-L4. The pharyngeal armature and piercing mandibles suggest a generalist feeder and opportunistic predator. Egg ansullae were randomly distributed (as opposed to rows in most other Culicoides spp.) and averaged 1.98 ansullae per 25 µm2 of egg surface. Ansullae were widened and porous apically, averaging 1.29 microns long, 0.65 µm at the base, and 1.06 µm at the apex. Only the L1 had a proleg, structurally complex, and featuring five rows of posteriorily directed, shorter spines (1-3 µm long) and 8-10 setae 18 µm long with hooked tips. On terrestrial mud, this proleg was used for surface crawling, and the L1 navigated dry conditions better than the more aquatic L2-L4 stages. Eggs are laid above waterline, and the L1 proleg is probably an advantage in terrestrial surface movement.

Commercially available formulations of two entomopathogenic fungi, Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Clavicipitaceae) and Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae), were assessed for control of Culex quinquefasciatus Say (Diptera: Culicidae) in underground storm drain systems (USDS) in the Coachella Valley of southern California. Each of three treatments, the two fungi or a water control, was applied to 1 m2 of vertical wall at eight USDS sites in spring and autumn of 2015. Fungal infectivity and lethality were assessed at 1 d and 1, 2, and 4 wk post-application. Overnight bioassays using adult lab-reared female mosquitoes were carried out on the treated USDS wall areas and then mosquitoes were held in the laboratory for up to 21 d to allow fungal infections to be expressed. Postmortem fungal sporulation was assessed up to 2 wk at 100% humidity. Mosquito-fungal interactions also were assessed in bioassays of the three treatments on filter paper exposed to USDS conditions during autumn. Metarhizium anisopliae killed mosquitoes faster than B. bassiana; nevertheless, both freshly applied formulations caused greater than 80% mortality. Fungal persistence declined significantly after 1 wk under USDS conditions, but some infectivity persisted for more than 4 wk. Beauveria bassiana was more effective against Cx. qinquefasciatus in the spring, while M. anisopliae was more effective in the cooler conditions during autumn. USDS environmental conditions (e.g., temperature, relative humidity, standing water) influenced fungal-related mortality and infection of Cx. quinquefasciatus. The utility of these fungal formulations for mosquito abatement in the Coachella Valley and implications for fungal control agents in USDS environments are discussed.

Insecticidal fly baits are important tools for adult house fly, Musca domestica L. (Diptera: Muscidae), control, especially on animal operations. Two house fly baits, containing either cyantraniliprole or dinotefuran, were tested on a dairy farm for attractiveness over time compared to a sugar control. Sticky trap and bucket trap house fly catches were recorded for each bait type at 1 h, 3 h, 6 h, 12 h, 24 h, 48 h, 72 h, and 168 h. After 1 wk of exposure to flies and field conditions, these 'aged' baits were tested against fresh baits for fly visitation in the field over 1 h. House flies from each bait type (aged and fresh) were collected and kept under laboratory conditions to assess mortality over 3 d. Average visitation of individual flies to each bait type (fresh) in the field was also evaluated. Sticky traps did not show significant fly catch differences among bait types over time, however bucket trap catches did show significant differences for cyantraniliprole bait and dinotefuran bait compared to sugar at 72 h and 168 h. No significant differences among fly visitation to aged or fresh baits were found. Fresh cyantraniliprole bait and dinotefuran bait resulted in greater fly mortality compared to controls, but not compared to aged toxic baits. Average house fly visitation time was greatest for sugar and cyantraniliprole bait.

California pastured cattle were treated with 250 ml of a 15% mixture of fatty acids (C8-C9-C10) or 125 ml of 2% geraniol in a mineral oil carrier to assess impacts on horn flies, Haematobia irritans (L.) (Diptera: Muscidae) over two summers. Horn flies were netted from cattle every 3-4 d for 2 wk before treatment, 2 wk during treatment (four treatments, with flies collected before each treatment), and 2 wk after treatments ceased. Blood meal weights were estimated by hemoglobin assay of excised abdomens. Other females were dissected to determine the number of active ovarioles and the stage of primary follicle development. Depending on year and herd, pretreatment males contained an average of 0.6-1.0 mg of blood, while females contained 1.7-2.7 mg. Pretreatment egg development (least developed oocytes were stage 1 and fully developed eggs were stage 5) averaged 3.7-4.3, and number of active ovarioles averaged 18.1 to 19.6/female. During treatment periods, significant reductions in blood weight were noted for females, but usually not for males, and females also often exhibited reduced mean oocyte stage and number of active ovarioles. Peaks in proportions of young nulliparous females (oocyte stages 1 or 2) were seen during some repellent application periods. This suggested older females had been killed or driven off from the local population by the treatments, and flies on cattle included more young flies that likely were recent arrivals. The repellent-oil mixture thus impacted blood feeding, reproductive fitness, and probably age structure in the field.

Light-baited suction traps are one of the most widely used tools for vector surveillance. Their popularity stems from ease of use even in remote locations, range and abundance of species caught, and low cost. The availability of smaller, portable models, like the CDC miniature light trap, have further increased their ubiquity in entomological field studies. However, when researchers have looked, light trap collections are usually biased in ways that may affect data interpretation for epidemiological studies. If used alone, light traps may fail to collect important or infected vectors, and light traps are inefficient or ineffective when competing ambient light is present. In this article, we discuss these biases and limitations in terms of their effect on collection efficiency, population data, and pathogen detection. While light trap data certainly have a purpose, an over-reliance on light trapping risks drawing false conclusions about vector populations and vector-borne disease epidemiology. These concerns are especially troubling when light trap data are used to inform policy decisions meant to protect human and animal health. Particularly when a species' response to light is unknown or poorly characterized, light traps should be used in conjunction with supplemental sampling methods. Researchers conducting vector surveillance field studies should carefully consider their study design and objectives when deciding on a trapping method or methods, and specifically endeavor to understand the limitations of their data. Only then can researchers take advantage of the best attributes of light traps while avoiding their dark side.

Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae) is a key bluetongue virus vector in the United States. Immatures occur in mud near the edges of wastewater ponds and are understudied targets for control efforts. Eggs of C. sonorensis were collected in the morning from a dairy wastewater pond bank by taking 5-ml surface mud samples along four transects on each of six dates. Surface mud samples parallel to waterline (10-cm long, 1-cm wide, and 0.5-cm deep) were removed at 5-cm increments ranging from 15 cm below waterline up to 25 cm above waterline. Eggs were removed using MgSO4 flotation, held on moist filter paper, and scored for hatching over 3 d. Eggs hatching on days 2 and 3 were assumed to have been laid on the test night. Water levels were stable within a night according to time-lapse camera photos. Most samples from below the waterline had no eggs and were not analyzed statistically. Mean (±SE) sample moisture (25.8 ± 2.1 at 5 cm above waterline and 19.8 ± 2.6% at 25 cm above waterline) did not vary significantly by position above waterline. The highest density of viable eggs (21 eggs/5 ml), proportion of mud samples positive for viable eggs (75%), and proportion of eggs hatching (80%) were found 5 cm above waterline. Oviposition in the few hours after sunset is adaptive, allowing eggs to age, develop the serosal cuticle, and resist later desiccation. As a potential control method, reducing water levels after midnight would encourage young egg desiccation.

The northern fowl mite, Ornithonyssus sylviarum (Canestrini & Fanzago, 1877), is found on several continents and has been a major pest of poultry in the United States for nearly a century. Lack of earlier USA reports in the United States suggests an introduction or change to pest status in domestic poultry systems occurred in the early 1900s. Though predominantly a nest-parasite of wild birds, this obligate hematophagous mite is a permanent ectoparasite on domestic birds, especially egg-laying chickens. Economic damage is incurred by direct blood feeding and activation of the of host's immune responses. This in turn causes decreased egg production and feed conversion efficiency, and severe infestations can cause anemia or death to birds. Here we review the biology, ecology, and recent control measures for the northern fowl mite. Photomicrographs are included of adult males and females, protonymphs, and larvae with key characters indicated. Special emphasis is placed on current knowledge gaps of basic and applied science importance., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Adult horn fly populations were tracked on cattle for 2-week periods before, during and after multiple treatments (every 3-4days) with two repellents in a mineral oil carrier. Cattle were sprayed four times in a two-week period either with 2% geraniol (125ml/cow) or a 15% mixture of short chain fatty acids (C8-C9-C10)(250ml/cow), and there were untreated control cattle. Trials were conducted in California and North Carolina for 3 summers. Short-term fly counts (same day) on treated cattle were reduced by 61-99%, depending on material and trial, and the fatty acid mixture provided better control than geraniol. Horn fly counts were suppressed for 1-3 d and rebounded somewhat after both treatments. Consecutive treatments showed evidence of persistent impact in California where herds were more isolated. Rebounds to pre-treatment levels 3-4 d after treatment occurred more often in North Carolina, where other infested cattle were closer to treated herds. By 3-4 d post-treatment, horn flies were reduced by 29-61% in California and 0-83% in North Carolina, relative to pre-treatment. Background behavior frequencies were assessed from hundreds of counts on untreated, infested California cattle, where horn flies were the only abundant biting fly. Behavior averages were 16.5 tail flicks, 7.6 skin twitches, 1.2 head throws, or 0.2 leg stamps per 2min observation period. At horn fly densities from about 200 to more than 1000 flies per animal (moderate to high numbers), fly defensive behaviors on control cattle were poorly related (or unrelated) to fly numbers. Immediately after repellent application, however, flies were almost absent and behavior frequencies dropped distinctly. Cattle fly defensive behaviors therefore seem to be quite sensitive to low (less than 100 flies/animal) horn fly densities, and behaviors would be a poor quantitative tool to track fly stress at moderate densities and above. Both geraniol and the fatty acids show promise for horn fly control, especially in organic agriculture. Treatments at 1-2 d intervals probably would keep infestations below the economic threshold (200 flies/cow)., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Hovering and swarming activity of male Tabanus calens L. were documented at distinctive sites above a woodland road clearing in Knox Co., TN (7 d), and above a slow-flowing river in Morris Co., NJ (6 d). Activity was restricted to the period near sunset. Mean activity typically began about 10 min before sunset and continued until about 10 min after sunset, with an average duration of 18.7 min (Tennessee) and 20.8 min (New Jersey). Swarming began earlier and thus lasted longer (55 min) on one cool New Jersey date. Onset and particularly cessation of activity tended to be statistically correlated with sunset time. Most males hovered at heights of 5-8 m in Tennessee, and they hovered in stable locations for a median of only 10 s (maximum 2 min 57 s) before darting away. Hovering in New Jersey was observed from 0.5-8.0 m above the water. The behavior of T. calens might be especially interesting to study in more detail, given its unusual eye morphology (lack of obvious dorsoventral facet size differences found in males of most other Tabanidae)., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background: Northern fowl mites (Ornithonyssus sylviarum) are obligate hematophagous ectoparasites of both feral birds and poultry, particularly chicken layers and breeders. They complete their entire life-cycle on infested birds while feeding on blood. Infestations of O. sylviarum are difficult to control and resistance to some chemical classes of acaricides is a growing concern. The contact susceptibility of O. sylviarum to a new active ingredient, fluralaner, was evaluated, as well as other compounds representative of the main chemical classes commonly used to control poultry mite infestations in Europe and the USA., Methods: Six acaricides (fluralaner, spinosad, phoxim, propoxur, permethrin, deltamethrin) were dissolved and serially diluted in butanol:olive oil (1:1) to obtain test solutions used for impregnation of filter paper packets. A carrier-only control was included. Thirty adult northern fowl mites, freshly collected from untreated host chickens, were inserted into each packet for continuous compound exposure. Mite mortality was assessed after incubation of the test packets for 48 h at 75% relative humidity and a temperature of 22 °C., Results: Adult mite LC 50 /LC 99 values were 2.95/8.09 ppm for fluralaner, 1587/3123 ppm for spinosad, 420/750 ppm for phoxim and 86/181 ppm for propoxur. Permethrin and deltamethrin LC values could not be calculated due to lack of mortality observed even at 1000 ppm., Conclusions: Northern fowl mites were highly sensitive to fluralaner after contact exposure. They were moderately sensitive to phoxim and propoxur, and less sensitive to spinosad. Furthermore, the tested mite population appeared to be resistant to the pyrethroids, permethrin and deltamethrin, despite not being exposed to acaricides for at least 10 years.

Culicoides sonorensis Wirth and Jones biting midges are the primary North American vectors of bluetongue virus (BTV), which infects domestic ruminant livestock and can cause high morbidity and mortality. Both virus and vector exhibit highly seasonal activity patterns, even in subtropical climates like southern California. However, the exact mechanism of BTV and Culicoides overwintering has been debated. In this study, we examined the supercooling point (SCP) and lower lethal temperature of a laboratory colony of C. sonorensis eggs, larvae, and pupae, as well as of field-collected larvae and pupae from subtropical (California) and temperate (Colorado) climates. Larvae and pupae succumbed to temperatures higher than their respective SCPs, indicating death from prefreezing cold injury. Eggs were the most cold-tolerant life stage, and were able to tolerate 1-h exposures to temperatures as low as -20 °C without suffering complete mortality. Larvae were the least cold-tolerant life stage and suffered complete mortality at < -4 °C, while temperatures of -9 to -10 °C were required to kill all pupae. Larvae and embryos suffered chronic cold injury after exposure to subzero temperatures, which increased mortality. Field-collected larvae succumbed to slightly higher temperatures ( > -3 °C) than colony larvae, but did not differ significantly in their survival. Culicoides sonorensis immatures did not cold harden when exposed to 4 °C for 24 h before freezing. Results of this study indicate that C. sonorensis eggs are more tolerant of environmental stress than previously thought. Further work is needed on C. sonorensis winter survival in the field to corroborate laboratory studies., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Northern fowl mite management on conventionally caged birds relies on synthetic pesticide sprays to wet the vent. Cage-free chickens cannot be effectively treated this way, and pesticide use is restricted in organic production. Dustbathing behavior is encouraged in newer production systems for increased hen welfare. Diatomaceous earth (DE) is an approved organic insecticide that can be mixed with sand in dustboxes, suppressing mites but not excluding them, and potentially allowing development of mite immunity. We tested two hypotheses: 1) that DE-filled dustboxes placed before northern fowl mite introduction (prophylactic use) prevents mite populations from reaching economically damaging thresholds, and 2) that bird exposure to low mite numbers allows for protective hen immunity to develop and suppress mites after dustboxes are removed. We also tested if different beak trimming techniques (a commercial practice) affect mite growth. Mites were introduced to birds after dustboxes were made available. Average mite densities in flocks remained below damaging levels while dustboxes were available. Average mite populations rebounded after dustbox removal (even though DE persisted in the environment) regardless of the timing of removal. Mite densities on birds where a traditional hot-blade beak trimming technique was used (trial 1) were high. Mite densities in trial 2, where a newer precision infra-red trimming was used, were lower. The newer infra-red trimming method resulted in nearly intact beaks, which were better for mite control by bird grooming behaviors. The combination of early dustbox use and infra-red beak trimming should allow producers to avoid most mite damage in cage-free flocks., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The northern fowl mite, Ornithonyssus sylviarum, is the most damaging ectoparasite on egg-laying hens in the United States. One potential strategy for management is breeding for mite resistance. Genes of white leghorn chickens linked to the major histocompatibility complex (MHC) were previously identified as conferring more (B21 haplotype) or less (B15 haplotype) mite resistance. However, immune responses can be energetically costly to the host and affect the economic damage incurred from mite infestations. We tested energy costs (resting metabolic rate) of mite infestations on egg-laying birds of both MHC B-haplotypes. Resting metabolic rates were documented before (pre-) mite infestation, during (mid-) infestation, and after peak (late) mite infestation. Mite scores, economic parameters (egg production, feed consumption), and physiological aspects such as skin inflammation and skin temperature were recorded weekly. Across experiments and different infestation time points, resting metabolic rates generally were not affected by mite infestation or haplotype, although there were instances of lower metabolic rates in infested versus control hens. Skin temperatures were recorded both at the site of mite feeding damage (vent) and under the wing (no mites), which possibly would reflect a systemic fever response. Ambient temperatures modified skin surface temperature, which generally was not affected by mites or haplotype. Feed conversion efficiency was significantly worse (4.9 to 17.0% depending on trial) in birds infested with mites. Overall egg production and average egg weight were not affected significantly, although there was a trend toward reduced egg production (2 to 8%) by infested hens. The MHC haplotype significantly affected vent skin inflammation. Birds with the mite-resistant B21 haplotype showed earlier onset of inflammation, but a reduced overall area of inflammation compared to mite-susceptible B15 birds. No significant differences in resting energy expenditure related to mite infestation or immune responses were detected. Potential breeding for resistance to mite infestation using these two haplotypes appears to be neutral in terms of impact on hen energy costs or production efficiency, and may be an attractive option for future mite control., (© 2016 Poultry Science Association Inc.)

Animal welfare-driven legislation and consumer demand are changing how laying chickens are housed, thus creating challenges for ectoparasite control. Hens housed in suspended wire cages (battery cages) are usually treated with high-pressure pesticides. This application type is difficult in enriched-cage or cage-free production. Alternatives to pesticide sprays are needed in enriched-cage or cage-free systems. In this study, we tested the efficacy of sulfur dust deployed in "dust bags" for control against the northern fowl mite (Ornithonyssus sylviarum), which causes host stress, decreased egg production, and reduced feed conversion efficiency. Dust bags were hung from the tops of cages or were clipped to the inside front of cages. We also tested permethrin-impregnated plastic strips, marketed for ectoparasite control in caged or cage-free commercial and backyard flocks. Previous work has shown sulfur to be very active against poultry ectoparasites; however, we found that the placement of bags was important for mite control. Sulfur in hanging bags reduced mites on treatment birds by 95 or 97% (depending on trial) within one week of being deployed, and mite counts on these birds were zero after 2 wk. Clipped sulfur bags acted more slowly and did not significantly reduce mites in one trial, but reduced mite counts to zero after 4 wk in trial 2. Permethrin strips had no effect on mite populations. This may have been due to mite resistance, even though this mite population had not been exposed to pyrethroids for several years. Sulfur bags should be effective in caged or cage-free systems., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Culicoides sonorensis Wirth & Jones (Diptera: Ceratopogonidae) is the primary North American vector of bluetongue virus (BTV), which can cause high morbidity and mortality in ruminant livestock or wildlife. Worldwide, most Culicoides surveillance relies on light (usually UV) traps typically placed near animals or larval development sites. However, the trapping method can cause sex, species and parity biases in collections. We collected C. sonorensis from three dairies in California using suction traps baited with CO2 , UV light or CO2  + UV placed near animals, wastewater ponds, or in fields. Higher numbers of parous females were collected using CO2  + UV traps, although this difference was only significant on one dairy. UV traps were poor at collecting nulliparous females, but the addition of UV to a trap increased the abundance of males in a collection. Traps set in open fields collected significantly higher numbers of males and females than in either of the other two locations. In some cases, there was a significant interaction between the trap type and site. We discuss the limitations of traditional trapping methodologies for C. sonorensis and make suggestions for vector surveillance., (© 2016 The Royal Entomological Society.)

Horn flies [Haematobia irritans (Diptera: Muscidae) (L.)] and face flies [Musca autumnalis (Diptera: Muscidae) De Geer] use the same larval resource, but their interactions are poorly studied. Dung pats (n = 350) were core sampled in the summers of 2012 and 2013 from irrigated pastures in Pomona, California, U.S.A. (34°03'N, 117°48'W) and held for face fly and horn fly emergence. Surface areas and estimated weights were recorded for each whole pat. Almost half (42.0%) of the pat cores yielded neither fly, 29.7% yielded horn flies only, 12.9% yielded face flies only and 15.4% yielded both flies. Of the fly-positive pats, surface area and mass were larger for face fly-occupied pats, whereas horn fly-occupied pats were smaller. Pats shared by the two species were intermediate. Horn flies per positive core were unaffected by the absence/presence of face flies, but half as many face flies emerged when pats were co-inhabited by horn flies. Face flies inhabited larger pats, which might better resist heating and drying, to which they are susceptible; horn flies inhabited a broad pat size range. Horn fly tolerance of lower dung moisture probably allows horn flies to colonize and survive in a wide range of pats in dry areas like southern California., (© 2016 The Royal Entomological Society.)

Horn flies, Haematobia irritans (L.), were photographed on cattle and then immediately estimated visually in the field on pastured beef cattle in southern California. Horn flies were counted in the pictures later on a computer screen. For 479 counts on individual cattle, the concordance correlation coefficient between the visual and photo-based assessments was 0.790, but was better for the higher half of the counts (0.732) than for the lower half of the counts (0.217). Major axis regression indicated that visual estimates were consistently higher than the number of flies counted in the pictures across the entire density range. Based on average raw means, the visual estimates averaged 21% higher than the photo counts. Visual estimates included flies on the belly and lower legs that could not be seen in a photo, and lower densities may have been more susceptible to such error. Where flies can be seen well, e.g. for very tame animals, the visual estimates were much faster and more cost-efficient and were sufficient to track relative horn fly abundance changes., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Peridomestic ("backyard") chicken flocks are gaining popularity in the developed world (e.g., North America or Europe), yet little is known regarding prevalence or severity of their ectoparasites. Therefore, five birds on each of 20 properties throughout southern California were surveyed in summer for on-host (permanent) and off-host dwelling (temporary) ectoparasites. Only four premises (20%) were entirely free of ectoparasites. In declining order of prevalence (% of premises), permanent ectoparasites included six chicken louse species: Menacanthus stramineus (Nitzsch) (50%), Goniocotes gallinae (De Geer) (35%), Lipeurus caponis (L.) (20%), Menopon gallinae (L.) (15%), Menacanthus cornutus (Schömmer) (5%), and Cuclotogaster heterographus (Nitzsch) (5%). Only one flea species, Echidnophaga gallinacea (Westwood) (20%), was found. Three parasitic mite species were observed: Ornithonyssus sylviarum (Canestrini & Fanzago) (15%), Knemidocoptes mutans (Robin & Lanquetin) (10%), and Dermanyssus gallinae (De Geer) (5%). Many infestations consisted of a few to a dozen individuals per bird, but M. stramineus, G. gallinae, M. cornutus, and E. gallinacea were abundant (dozens to hundreds of individuals) on some birds, and damage by K. mutans was severe on two premises. Off-host dwelling ectoparasites were rare (D. gallinae) or absent (Cimex lectularius L., Argasidae). Parasite diversity in peridomestic flocks greatly exceeds that is routinely observed on commercial chicken flocks and highlights a need for increased biosecurity and development of ectoparasite control options for homeowners., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The face fly (Musca autumnalis De Geer) and horn fly (Haematobia irritans ([L.])) were studied at the southern edge of the face fly's North American range, examining southern California geographic distribution, seasonal activity on cattle and in dung, and diapause. Face flies were common only at Pomona (34°03'N, 117°48'W). Other irrigated pastures, even those only slightly inland from Pomona, were probably too warm for face flies, due to a steep west (cooler) to east (warmer) temperature gradient. Horn flies were abundant at all sites. Adult densities on cattle, adults emerging from dung pats, and prevalence of fly-positive pats were assessed for both fly species throughout a year at Pomona. Summer adult horn fly densities of 500-2,000 flies per cow, or face fly densities of 3-10 flies per face, were common. Summer prevalence of face fly-positive pats and horn fly-positive pats was about 20-40% and 30-70%, respectively. Face fly adults diapaused from late October until late March and early April. Horn flies probably diapaused as pupae from late October or early November to early-mid March, although some emerged in winter. Experimental cohorts of October-emerging adult face flies were held in a representative overwintering site. They exhibited hypertrophied fat body and undeveloped oocytes, which are characteristics of diapause, and survived until the following spring. The dominant diapause cues in face and horn flies are probably photoperiod and temperature. Despite warm winter temperatures that would permit activity of both species, and despite relatively long winter day lengths, face flies and most horn flies still diapaused at this latitude., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The Bluetongue and Schmallenberg virus outbreaks in Europe between 1999 and 2013 stimulated progress in Culicoides field ecology and control. In responding to these outbreaks, however, the deficit of broadly trained entomologists became evident. Culicoides research has been uneven, and critical gaps remain in our knowledge of vector ecology. Certain aspects of adult midge biology have gotten more attention (biting rates and relationship to trap collections, seasonal activity and survey, taxonomy of critical species complexes). Some aspects of adult midge biology (survival, resting site selection, or direct dispersal measurements) have been understudied. More work is needed on Culicoides immatures (taxonomy and details of ecology). Control research has centred on insecticide treatments applied to animals. However, our true goal is not vector control, but reducing disease agent transmission. We require field vector control targets (e.g. how far we might need to reduce biting rates) needed to interrupt transmission. The most critical need is for studies incorporating vector control experimentally into epidemiological studies in active arbovirus transmission zones. Such field interdisciplinary studies are absolutely required to understand transmission dynamics. Through the power of experimental field studies, we should be able to develop both theoretical and operational guidelines for disease management.

Background: Pathogen manipulation of host behavior can greatly impact vector-borne disease transmission, but almost no attention has been paid to how it affects disease surveillance. Bluetongue virus (BTV), transmitted by Culicoides biting midges, is a serious disease of ruminant livestock that can cause high morbidity and mortality and significant economic losses. Worldwide, the majority of surveillance for Culicoides to assess BTV transmission risk is done using UV-light traps. Here we show that field infection rates of BTV are significantly lower in midge vectors collected using traps baited with UV light versus a host cue (CO2)., Methods: We collected Culicoides sonorensis midges in suction traps baited with CO2, UV-light, or CO2 + UV on three dairies in southern California to assess differences in the resulting estimated infection rates from these collections. Pools of midges were tested for BTV by qRT-PCR, and maximum likelihood estimates of infection rate were calculated by trap. Infection rate estimates were also calculated by trapping site within a dairy. Colonized C. sonorensis were orally infected with BTV, and infection of the structures of the compound eye was examined using structured illumination microscopy., Results: UV traps failed entirely to detect virus both early and late in the transmission season, and underestimated virus prevalence by as much as 8.5-fold. CO2 + UV traps also had significantly lower infection rates than CO2-only traps, suggesting that light may repel infected vectors. We found very high virus levels in the eyes of infected midges, possibly causing altered vision or light perception. Collecting location also greatly impacts our perception of virus activity., Conclusions: Because the majority of global vector surveillance for bluetongue uses only light-trapping, transmission risk estimates based on these collections are likely severely understated. Where national surveillance programs exist, alternatives to light-trapping should be considered. More broadly, disseminated infections of many arboviruses include infections in vectors' eyes and nervous tissues, and this may be causing unanticipated behavioral effects. Field demonstrations of pathogen-induced changes in vector behavior are quite rare, but should be studied in more systems to accurately predict vector-borne disease transmission.

The effects of beak condition on ectoparasite populations and preening in laying hens were investigated. Beak-trimmed and beak-intact caged Hy-Line W-36 hens were infested with either chicken body lice or northern fowl mites using a 2×2 factorial design with 4 replicate cages (each containing 2 hens)/treatment: 1) BTL: (beak-trimmed lice-infested); 2) BTM: (beak-trimmed mite-infested); 3) BIL: (beak-intact lice-infested); and 4) BIM: (beak-intact mite-infested). Mite scores and lice numbers were estimated weekly. Hens were video recorded the wk before infestation and at wk 6 and 9 post-infestation. Time spent preening on 6 body areas and in total were analyzed using a repeated measures ANOVA. There was a wk×beak condition interaction for lice loads, with BTL harboring approximately 17 times more lice than BIL from wk 7 to 10 post-infestation (P<0.0001). Beak condition affected mite loads (P<0.0001), with BTM having a higher mite score (3.8±0.26) than BIM (1.4±0.26). At peak infestation, BTL spent more total time preening (P=0.02, s±SE: 232.1±37.6) than prior to infestation (33.9±37.6) and directed their preening behavior towards the vent. In contrast, BIL (73.9±37.6), BTM (9.4±1.6), and BIM (8.6±1.6) did not increase total time spent preening over pre-infestation levels (103.6±37.6, 5.8±1.6, 6.7±1.6 respectively), although BTM did redirect their preening behavior toward the vent. This study confirmed previous studies showing that an intact beak is important for reducing ectoparasite infestations. Preening behavior increased in response to lice infestation, but only in beak-trimmed hens; preening behavior and louse load were correlated at peak infestation. In contrast, mite infestation did not lead to increased preening, and there was no correlation between preening and mite load. However, both lice- and mite-infested hens directed preening behavior predominantly towards the vent where these parasites are typically found., (© 2015 Poultry Science Association Inc.)

Culicoides biting midges (Diptera: Ceratopogonidae) are biological vectors of two important viruses impacting North American ruminants--bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV). These viruses have been identified for over 60 years in North America, but we still lack an adequate understanding of the basic biology and ecology of the confirmed vector, Culicoides sonorensis, and know even less about other putative Culicoides vector species. The major gaps in our knowledge of the biology of Culicoides midges are broad and include an understanding of the ecology of juveniles, the identity of potential alternate vector species, interactions of midges with both pathogens and vertebrates, and the effectiveness of potential control measures. Due to these broad and numerous fundamental knowledge gaps, vector biologists and livestock producers are left with few options to respond to or understand outbreaks of EHD or BT in North America, or respond to emerging or exotic Culicoides-transmitted pathogens. Here we outline current knowledge of vector ecology and control tactics for North American Culicoides species, and delineate research recommendations aimed to fill knowledge gaps.

Background: Toxic fly baits are commonly used for fly control in California animal operations. However, resistance development has been a problem. Comprehensive laboratory and field studies were conducted to test commercial baits (imidacloprid, methomyl, dinotefuran, spinosad) and one novel cyantraniliprole bait. A susceptible Musca domestica strain was compared with wild-type M. domestica and Fannia canicularis strains in the laboratory using choice/no-choice tests. Field visitation to baits and both short- and longer-term mortality were documented., Results: Susceptible Musca suffered high mortality with all baits after 3 days of choice and no-choice tests. Wild-type Musca mortality was more variable and higher in no-choice relative to choice tests. Fannia were most susceptible to spinosad > dinotefuran = cyantraniliprole > methomyl = imidacloprid. Field Musca were most attracted to spinosad > cyantraniliprole > dinotefuran > sugar > methomyl > imidacloprid. Delayed mortality from bait-fed field flies (captured and held with untreated food and water for 3 days) was ranked spinosad = cyantraniliprole > dinotefuran = methomyl > imidacloprid > sugar., Conclusion: Behavioral resistance of M. domestica to imidacloprid and methomyl persists. Spinosad and cyantraniliprole baits (delayed mortality) performed best. Speed of action may be a factor in use and misuse of baits., (© 2014 Society of Chemical Industry.)

The northern fowl mite, Ornithonyssus sylviarum (Canestri & Fanzago), is an ectoparasite of more than 70 species of North American wild birds, but it has a particularly significant impact on chickens, where it is a permanent resident of vent feathers. Improved control practices depend on a better understanding of host-mite relationships. ISA Brown hens were inoculated experimentally with northern fowl mite adults, and northern fowl mite populations developed naturally. Using a fast-response microprobe, temperatures of individual vent feathers (n = 15) were recorded at 5-mm increments along the length of the feather shaft. Immediately after temperatures were recorded, the individual feathers were quickly clipped at the skin surface and then flash-frozen between 2 small blocks of dry ice, freezing all northern fowl mite stages in situ. The feathers then were cut into 5-mm sections for careful mite enumeration by life stage. There were no overall differences among life stages in the distributions on the feather. Mite positions on feathers (distance from skin) varied distinctly with feather zone temperatures, as well as with ambient and average temperatures over the prior 24 hr. Ambient temperature at time of sampling affected the positions of the 2 mobile categories, adults and larvae/nymphs, but showed no statistical relationship with egg distribution. In contrast, ambient 24-hr temperature influenced the positions of all life stages. On-host feather temperatures reflected ambient temperatures. Feathers collected on hot days (ambient temperatures of 23-33 C) provided a narrow and quite warm range of temperature conditions for mites (often >30-36 C). Temperatures on cool days (ambient temperatures of <23 C) provided much wider on-host temperature ranges for mites to occupy (13-35 C). Mites were farther from the skin on warmer days. When mites had a broad range of temperatures, the feather temperature zone occupied by all life stages averaged 28-29 C. Mites move to occupy favorable temperature conditions on-host. When further out on feathers in warm weather, and under thermal stress, northern fowl mites either move off host or are dislodged. They then become a human pest, are noticed by farmers, and are more likely to disperse.

Culicoides midges are abundant hematophagous flies that vector arboviruses of veterinary and medical importance. Dramatic changes in the epidemiology of Culicoides-borne arboviruses have occurred since 1998, including the emergence of exotic viruses in northern temperate regions, increases in global disease incidence, and enhanced virus diversity in tropical zones. Drivers may include changes in climate, land use, trade, and animal husbandry. New Culicoides species and new wild reservoir hosts have been implicated in transmission, highlighting the dynamic nature of pathogen-vector-host interactions. Focusing on potential vector species worldwide and key elements of vectorial capacity, we review the sensitivity of Culicoides life cycles to abiotic and biotic factors. We consider implications for designing control measures and understanding impacts of environmental change in different ecological contexts. Critical geographical, biological, and taxonomic knowledge gaps are prioritized. Recent developments in genomics and mathematical modeling may enhance ecological understanding of these complex arbovirus systems.

ImageJ is an open-source software tool used for a variety of scientific objectives including cell counting, shape analysis and image correction. This technology has previously been used to estimate mosquito abundance in surveillance efforts. However, the utility of this application for estimating abundance or parity in the surveillance of Culicoides spp. (Diptera: Ceratopogonidae) has not yet been tested. Culicoides sonorensis (Wirth and Jones), a biting midge often measuring 2.0-2.5 mm in length, is an economically important vector of ruminant arboviruses in California. Current surveillance methods use visual sorting for the characteristics of midges and are very time-intensive for large studies. This project tested the utility of ImageJ as a tool to assist in gross trap enumeration as well as in parity analysis of C. sonorensis in comparison with traditional visual methods of enumeration using a dissecting microscope. Results confirmed that automated counting of midges is a reliable means of approximating midge numbers under certain conditions. Further evaluation confirmed accurate and time-efficient parity analysis in comparison with hand sorting. The ImageJ software shows promise as a tool that can assist and expedite C. sonorensis surveillance. Further, these methods may be useful in other insect surveillance activities., (© 2014 The Royal Entomological Society.)

Culicoides sonorensis Wirth and Jones transmits bluetongue virus and develops in a variety of polluted mud habitats. Egg desiccation tolerance was tested by obtaining eggs of known age, drying them, and placing them back on wet substrate. Eggs 4-10 h old failed to hatch after 12 h of drying at 75% relative humidity (RH). Older eggs (28-34 h) survived severe desiccation and >50% water weight loss. They regained their water within ≈2 h of rehydration. Relative to control eggs, average egg hatch was reduced by 36% after 12 h of drying, 79% after 24 h, 91% after 36 h, and 97% after 48 h. Some embryos (1%) survived and hatched after 60 h of drying and water losses of nearly 60%. Eggs in specific 25-40 h age categories did not differ in hatch after a 12-h desiccation stress; critical embryo age to survive drying is between 10 and 24 h. Humidity gradients relieved desiccation stress, and eggs appeared to regain water from saturated RH conditions. Individual, gravid C. sonorensis oviposited in 1-liter containers with an artificial mud bank. If they laid eggs, 73% deposited them singly in lines ranging up to 5-6 cm in length (meanderers), while 27% laid eggs in clumps (dumpers). Eggs were positioned an average of 45 ± 12 mm back from waterline. Younger eggs, if laid in early evening, may not experience severe desiccation. Embryo recovery from such severe desiccation could be adaptive in ephemeral habitats where the species may have evolved., (© 2014 Entomological Society of America.)

Bluetongue virus (BTV) is the cause of an economically important arboviral disease of domestic and wild ruminants. The occurrence of BTV infection of livestock is distinctly seasonal in temperate regions of the world, thus we determined the dynamics of BTV infection (using BTV-specific real time reverse transcriptase polymerase chain reaction) among sentinel cattle and vector Culicoides sonorensis (C. sonorensis) midges on a dairy farm in northern California throughout both the seasonal and interseasonal (overwintering) periods of BTV activity from August 2012 until March 2014. The data confirmed widespread infection of both sentinel cattle and vector midges during the August-November period of seasonal BTV transmission, however BTV infection of parous female midges captured in traps set during daylight hours also was detected in February of both 2013 and 2014, during the interseasonal period. The finding of BTV-infected vector midges during mid-winter suggests that BTV may overwinter in northern California by infection of long-lived female C. sonorensis midges that were infected during the prior seasonal period of virus transmission, and reemerged sporadically during the overwintering period; however the data do not definitively preclude other potential mechanisms of BTV overwintering that are also discussed.

The Sacramento (northern Central) Valley of California (CA) has a hot Mediterranean climate and a diverse ecological landscape that is impacted extensively by human activities, which include the intensive farming of crops and livestock. Waste-water ponds, marshes, and irrigated fields associated with these agricultural activities provide abundant larval habitats for C. sonorensis midges, in addition to those sites that exist in the natural environment. Within this region, C. sonorensis is an important vector of bluetongue (BTV) and related viruses that adversely affect the international trade and movement of livestock, the economics of livestock production, and animal welfare. To characterize the seasonal dynamics of immature and adult C. sonorensis populations, abundance was monitored intensively on two dairy farms in the Sacramento Valley from August 2012- to July 2013. Adults were sampled every two weeks for 52 weeks by trapping (CDC style traps without light and baited with dry-ice) along N-S and E-W transects on each farm. One farm had large operational waste-water lagoons, whereas the lagoon on the other farm was drained and remained dry during the study. Spring emergence and seasonal abundance of adult C. sonorensis on both farms coincided with rising vernal temperature. Paradoxically, the abundance of midges on the farm without a functioning waste-water lagoon was increased as compared to abundance on the farm with a waste-water lagoon system, indicating that this infrastructure may not serve as the sole, or even the primary larval habitat. Adult midges disappeared from both farms from late November until May; however, low numbers of parous female midges were detected in traps set during daylight in the inter-seasonal winter period. This latter finding is especially critical as it provides a potential mechanism for the "overwintering" of BTV in temperate regions such as northern CA. Precise documentation of temporal changes in the annual abundance and dispersal of Culicoides midges is essential for the creation of models to predict BTV infection of livestock and to develop sound abatement strategies.

Musca autumnalis DeGeer were collected in the summer and fall of 2011 and 2012 from a beef cattle herd in southern California Visual counts of Musca spp. on cattle faces were documented, and sweep net samples of face flies and other Diptera were also collected from cattle faces. Face flies dominated in the net collections, and 5-30 flies per face were common between early July and October 2011. Adult female M. autumnalis were dissected and examined for the presence of the host-specific nematode Paraiotonchium autumnale (Nickle). Overall, 67 of 887 (7.6%) adult face fly females were parasitized. M. autumnalis' ability to survive in such a southerly latitude (34 degree N) could reflect the rather temperate weather (coastal effects) and frequently irrigated pastures at the experimental site in southern California Preliminary observations suggest that face flies disappear from cattle during winter, despite generally favorable temperatures for fly activity. This is a possible indication of diapause and should be examined further.

"Candidatus Cardinium hertigii" (Bacteroidetes) is a maternally inherited endosymbiont known from several arthropods. Its mechanisms for persistence in host populations are mostly reproductive manipulation, though it has been occasionally reported to improve fitness parameters in several hosts. In Culicoides (Diptera: Ceratopogonidae) biting midges, the prevalence of "Candidatus Cardinium" infection was documented as moderate, with no detectable sex bias. We therefore investigated whether "Candidatus Cardinium" affects important fitness parameters, such as survival and body size, in Culicoides imicola, a dominant vector species. Field-collected midges were trapped and analyzed for survival under different environmental conditions and antibiotic treatment, taking into account "Candidatus Cardinium" infection status and parity status (i.e., parous or nulliparous). Additionally, wing lengths were measured as a proxy parameter for body size and analyzed together with "Candidatus Cardinium" infection data. The findings revealed no difference in survival of Culicoides infected with "Candidatus Cardinium" and that of uninfected midges in both parity states and under all tested conditions: optimal, starvation, heat, and antibiotic treatment. Beyond survival, no wing length difference was found for "Candidatus Cardinium"-infected versus uninfected midges. In aggregate, these findings support our conclusion that "Candidatus Cardinium" does not have an overt effect on the survival and size of adult C. imicola midges. "Candidatus Cardinium" may affect immature stages or may alter adult reproductive performance.

Adult female Musca domestica L. were collected in 2004 and 2005 from dairies in California, Minnesota, and Georgia. Relative abundance of (Z)-9-tricosene (muscalure) among the dominant eight hydrocarbons was determined. Fly heads then were removed to quantify pterin levels and estimate fly age, abdomens were dissected to score gonotrophic development and parity (follicular relics), and spermathecae were examined for sperm. Daily survival was assessed using two estimates of time required to become gravid: laboratory-based degree-day (DD) estimates and estimates based on pterin values in field-collected flies matched to their stages of gonotrophic development. Among newly emerged females (oocyte stage 1) with detectable muscalure, it comprised < approximately 1.5% of cuticular hydrocarbons. In muscalure-positive flies, muscalure comprised a higher proportion of cuticular hydrocarbons in older flies from California and Minnesota (6-9% when gravid) versus flies from Georgia (<2% when gravid). Females mated in early-intermediate stages of egg development. Life expectancy, using laboratory-derived estimates of time needed to become gravid, ranged from 3.6 to 10.6 d. Using equivalent pterin-based time estimates, life expectancy ranged from 4.0 to 19.5 d. Mean DD ages (12 degrees C threshold) of gravid flies varied widely (53-95 DD) and were congruent with laboratory-based estimates (52-57 DD) in only 7 of 12 farm-year combinations. Thus, house flies under natural conditions often required more time to develop eggs than laboratory models would predict, extending daily survival estimates based on gonotrophic age by 11-74%.

Culicoides species (Diptera: Ceratopogonidae) belonging to the Obsoletus and Pulicaris groups are considered to be the main vectors of bluetongue virus (BTV) in non Mediterranean Europe. Selected terrestrial microhabitats (n=17) on a farm in northern Spain were sampled repeatedly over a year-long period and characterized for use by Culicoides species for immature development. Concurrent use of CDC light traps showed the presence of 37 species and 66,575 specimens of adult Culicoides. A total of 28 species and 11,396 individuals emerged from laboratory-maintained soil samples. Culicoides obsoletus and Culicoides scoticus (pooled as Obsoletus complex) were particularly abundant (comprising 58.6% and 74.5% of the total collections in light traps and emergence traps respectively). Potential key vectors of animal viruses (such as BTV) were found in two main terrestrial types of microhabitats. In the case of C. obsoletus, different types of manure (old and composted manure, manure mixed with organic matter, and fresh manure) produced most of the specimens. In contrast, larvae of C. scoticus and Culicoides lupicaris were associated with soil substantially comprised of rotting leaf litter that included the parasitic plant Lathraea clandestina. Several species, Culicoides festivipennis, Culicoides punctatus and Culicoides brunnicans, were very common in mud at pond margins. Indeed, pond microhabitats and runoff below barn rooflines supported the greatest species richness. In the pond habitat, 49.4% of Culicoides specimens emerged from mud at the water edge, as opposed to 50 cm above (32.4%) and 1 meter above waterline (18%). Similar species richness, but statistically significant differences in abundance, were observed among the four pond microhabitats. Overall, the majority of the specimens were found in the upper layer (0-3 cm), except in manure, where they preferred deeper layers (>6 cm). Maximum peaks of abundance occurred in both light traps and soil samples in summer months, whereas increased captures in autumn were noticed only in light traps. Both trapping systems failed to collect adult Culicoides midges in the coldest months of December, January and February. The literature on immature habitats of species suspected in BTV transmission in Europe, the Pulicaris group and particularly the Obsoletus group, is briefly reviewed., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Hen housing (cage or cage-free) did not impact overall abundances of northern fowl mites, Ornithonyssus sylviarum (Canestrini & Fanzago) (Acari: Macronyssidae), or chicken body lice, Menacanthus stramineus (Nitzsch) (Phthiraptera: Menoponidae). Cage-free hens received a dustbox with sand plus diatomaceous earth (DE), kaolin clay or sulphur. Weekly use varied from none to 100% of hens; 73% of hens used the dustbox at least once. Ectoparasite populations on dustbathing hens (users) were compared with those on non-user cage-free and caged hens. All materials reduced ectoparasites on user hens by 80-100% after 1 week of dustbox use. Diatomaceous earth and kaolin failed to reduce ectoparasites on non-user hens, and ectoparasites on user hens recovered after dustbox removal. A sulphur dustbox eliminated mites from all hens (including non-users) within 2-4 weeks. Residual sulphur controlled mites until the end of the experiment (up to 9 weeks), even after the dustbox was removed. Louse populations on hens using the sulphur dustbox were reduced in 1-2 weeks. Residual sulphur effects were less evident in lice, but the use of a sulphur dustbox by a higher proportion of hens extended louse control to all hens. This is the first experimental study to show that bird dustbathing in naturally and widely available dust materials (particularly kaolin) can suppress ectoparasites and thus the behaviour is probably adaptive., (© 2012 The Authors. Medical and Veterinary Entomology © 2012 The Royal Entomological Society.)

Bluetongue (BT) is an important viral disease of ruminants that is transmitted by hematophagous Culicoides midges. We examined the seasonal patterns of abundance and infection of Culicoides sonorensis at four dairy farms in the northern Central Valley of California to develop estimates of risk for bluetongue virus (BTV) transmission to cattle at each farm. These four farms were selected because of their similar meteorological conditions but varying levels of vector abundance and BTV infection of cattle. C. sonorensis midges were collected weekly at each farm during the seasonal transmission period, using three different trapping methods: traps baited with either carbon dioxide (CO(2)) alone or traps with CO(2) and UV light, and by direct aspiration of midges from sentinel cattle. Analysis of BTV-infected midges using group and serotype-specific quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) assays confirmed that BTV serotypes 10, 11, 13 and 17 are all present in the region, but that midge infection rates and the number of BTV serotypes circulating differed markedly among the individual farms. Furthermore, more serotypes of BTV were present in midges than in sentinel cattle at individual farms where BTV circulated, and the virus was detected at each farm in midges prior to detection in cattle. BTV infection rates were remarkably lower among female C. sonorensis midges collected by CO(2) traps with UV light than among midges collected by either animal-baited aspirations or in CO(2) traps without light. A subsample of female midges examined from each collection method showed no overall differences in the proportion of female midges that had previously fed on a host. Findings from this study confirm the importance of using sensitive surveillance methods for both midge collection and virus detection in epidemiological studies of BTV infection, which is especially critical if the data are to be used for development of mathematical models to predict the occurrence of BTV infection of livestock., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Culicoides biting midges (Diptera: Ceratopogonidae) are vectors of a variety of pathogens including African horse sickness virus (AHSV), a member of the family Reoviridae, genus Orbivirus. AHSV causes African horse sickness (AHS), an endemic disease of equids with an extremely high mortality rate in horses in sub-Saharan Africa. Culicoides (Avaritia) imicola Kieffer is considered to be the principal vector of AHSV and is the dominant Culicoides species in South Africa. Due to the global distribution of Culicoides vectors, there is a potential risk of AHS spreading from endemic areas to areas traditionally free of the disease, which could have a severe economical impact on the affected equine industry. As part of any risk assessment it is essential to monitor known vectors as well as potential vector species. In the present study, sampling of Culicoides insects was compared using overnight collections in the conventional Onderstepoort light trap and mechanical aspiration of midges at sunset from bait horses. Culicoides imicola was confirmed as the predominant species using both trapping methods. Other species, mainly Culicoides (Avaritia) bolitinos Meiswinkel and Culicoides (Avaritia) gulbenkiani Caeiro, were highly underrepresented in the light trap collections, but made a significant contribution to the mechanical aspiration catches. The time for optimal collection differed between the trapping methods, leading to the conclusion that mechanical aspiration is a useful addition to conventional light trap collection and possibly the better choice when investigating insect vectors. An infection rate of 1.14% was calculated for the midge population based on real-time quantitative reverse-transcription polymerase chain reaction (RT-qPCR) assays of collected Culicoides midges, which exceeds previous estimates. This is probably due to the increased sensitivity of the RT-qPCR assay used in this study as compared to the virus isolation assays used in previous studies. RT-qPCR-positive midges were present in midge pools obtained from both light trap and mechanical aspiration. Seven of the positive pools consisted of C. imicola only, four contained mixed species and one pool contained no C. imicola, suggesting the presence of AHSV in midges of other species., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Bluetongue is an economically important arboviral disease of ruminants that is transmitted by hematophagous Culicoides midges. In light of dramatic recent changes in the global distribution of bluetongue virus (BTV), the goals of this study were to re-evaluate the prevalence of BTV infection of cattle and abundance of Culicoides midges on individual dairy farms in California. A serosurvey of adult dairy cattle confirmed that BTV infection is prevalent throughout much of the state, although the coastal northwestern region remains free of infection and prevalence varies markedly among farms in the remainder of the state. Intensive sampling for one year of 4 farms in the northern Central Valley of California showed that the abundance of Culicoides midges was markedly different and coincided with the prevalence of BTV infection of sentinel cattle on each farm. Mean maximum and minimum temperatures and other meteorological parameters were similar on all 4 farms, thus we speculate that particular management practices were responsible for both the increased midge abundance and prevalence of BTV infection of cattle at individual farms. Specifically, it is concluded that variation in vector abundance at individual farms most likely is the result of waste-water lagoon and irrigation management practices, leading to higher BTV infection rates among livestock held on farms with more waste-water lagoons and greater acreage of land for waste-water irrigation., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Human sweat, liquid ammonia, and bovine blood are known to be attractive to some hematophagous flies. These materials were evaluated with and without carbon dioxide (CO(2)) for their ability to increase capture of female "canyon flies" (Fannia conspicua Malloch) using CDC-type suction traps (without light). Ammonia acted synergistically with CO(2) to increase trap catch 89.9% over CO(2) alone. There was no synergistic effect of human sweat or bovine blood with CO(2). In the absence of CO(2), none of the three materials increased trap catch of female canyon flies relative to non-baited traps. Implications for canyon fly control and further trap improvement are discussed., (© 2011 The Society for Vector Ecology.)

Objective: Ornithonyssus sylviarum (northern fowl mite) and Menacanthus stramineus (chicken body louse) are key poultry pests currently controlled by exclusion or pesticide application. We studied how host beak condition affected their populations over time and how the lice and mites might interact on a host., Methods: Beak-trimmed or beak-intact white leghorn hens were infested initially with either mites or lice and subsequently challenged using the alternate ectoparasite species (reciprocal transfer), while other hens harboured only the initial ectoparasite species., Results: Beak-trimmed hens had far higher ectoparasite numbers relative to beak-intact hens, and the 2 ectoparasites showed evidence of grooming-mediated competitive asymmetry. On beak-trimmed hens, larger numbers of lice quickly nearly completely excluded mites in competition for enemy-free space (lower abdomen), while in the reciprocal transfer mites did not affect louse numbers on beak-trimmed hosts. The 2 ectoparasites co-existed on beak-intact hens, which were better able to defend the lower abdomen habitat by grooming., Conclusion: Lice are somewhat less damaging and much easier to control relative to mites, and might be used to eliminate mites in commercial, beak-trimmed flocks. Beak trimming impairs host grooming and contributes greatly to the high ectoparasite populations seen in commercial flocks. The study adds incentives for poultry breeders to develop more docile hen strains that can be held without beak trimming. This has advantages both to welfare advocates and producers who may no longer need to use insecticides for pest control or be concerned about worker exposure to pesticides.