Parasite load effects on sex ratio, size, survival and mating fitness of Heleidomermis magnapapula in Culicoides sonorensis.

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 mm3 and females from 0.0121 - 0.1110 mm3. 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.

Hiding in Plain Sight: An Abundant and Widespread North American Horse Fly (Diptera: Tabanidae) in the Tabanus sulcifrons Group, Tabanus variegatus Fabricius, Redescribed.

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.

Collecting and Monitoring for Northern Fowl Mite (Acari: Macronyssidae) and Poultry Red Mite (Acari: Dermanyssidae) in Poultry Systems.

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.

Genetic Structure of Northern Fowl Mite (Mesostigmata: Macronyssidae) Populations Among Layer Chicken Flocks and Local House Sparrows (Passeriformes: Passeridae).

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.

Laboratory evaluation of stable isotope labeling of Culicoides (Diptera: Ceratopogonidae) for adult dispersal studies.

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 13C and 15N 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 13C or 15N enrichment levels. CONCLUSIONS: Stable isotope labeling is life-long, and does not interfere with natural insect behaviors. Stable isotope enrichment using 13C or 15N 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.

Sex Ratios and Mating Status of the Horn Fly on Pastured Cattle as a Function of Repellent-Oil Treatments.

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.

Longevity of Fly Baits Exposed to Field Conditions.

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.

Evaluation of Two Entomopathogenic Fungi for Control of Culex quinquefasciatus (Diptera: Culicidae) in Underground Storm Drains in the Coachella Valley, California, United States.

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.

Egg and Larval Morphology of Culicoides sonorensis (Diptera: Ceratopogonidae).

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.

The Dark Side of Light Traps.

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.

Field Distribution and Density of Culicoides sonorensis (Diptera: Ceratopogonidae) Eggs in Dairy Wastewater Habitats.

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.

Effects of Fatty Acid and Geraniol Repellent-Oil Mixtures Applied to Cattle on Blood Feeding and Reproductive Parameters in Field Populations of Haematobia irritans (Diptera: Muscidae).

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.

A review of the biology, ecology, and control of the northern fowl mite, Ornithonyssus sylviarum (Acari: Macronyssidae).

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.

Field trials of fatty acids and geraniol applied to cattle for suppression of horn flies, Haematobia irritans (Diptera: Muscidae), with observations on fly defensive behaviors.

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).

Comparative in vitro evaluation of contact activity of fluralaner, spinosad, phoxim, propoxur, permethrin and deltamethrin against the northern fowl mite, Ornithonyssus sylviarum.

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 LC50 /LC99 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.

Hovering and Swarming Behavior of Male Tabanus calens (Diptera: Tabanidae) in Tennessee and New Jersey, USA.

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).

Low Temperature Tolerance of Culicoides sonorensis (Diptera: Ceratopogonidae) Eggs, Larvae, and Pupae From Temperate and Subtropical Climates.

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.

Timing Diatomaceous Earth-Filled Dustbox Use for Management of Northern Fowl Mites (Acari: Macronyssidae) in Cage-Free Poultry Systems.

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.