New thinking for filth fly control: residual, non-chemical wall spray from volcanic glass.

Filth flies are of medical and veterinary importance because of the transfer of disease organisms to animals and humans. The traditional control methods include the use of chemical insecticides. A novel mechanical insecticide made from volcanic glass and originally developed to control mosquitoes (Imergard™ WP; ImG) was investigated for control of adult grey flesh flies, Sarcophaga bullata (Parker), secondary screwworms, Cochliomyia macellaria (F.), and house flies, Musca domestica L. In a modified WHO cone test device, the time to 50% mortality (LT50 ) when applied at 5 g/m2 (tested at 30 °C and 50% relative humidity (rH)) was 7.1, 4.3 and 3.2 h, respectively. When knockdown was included, the LT50 s were 5.5, 1.5 and 2.8 h, respectively. Application rates of 1.25 and greater g/m2 had the shortest LT50 s. The time to the LT50 increased for M. domestica as rH increased, but ImG was still active at the highest rH tested of 70%. Scanning electron micrographs showed ImG was present on all body parts, unlike that for mosquitoes where it was found mostly on the lower legs. These first studies on the use of Imergard WP against flies suggest this could be an alternative method for filth fly control.

Quantifying pteridines in the heads of blow flies (Diptera: Calliphoridae): Application for forensic entomology.

In forensic cases involving entomological evidence, establishing the postcolonization interval (post-CI) is a critical component of the investigation. Traditional methods of estimating the post-CI rely on estimating the age of immature blow flies (Diptera: Calliphoridae) collected from remains. However, in cases of delayed discovery (e.g., when remains are located indoors), these insects may have completed their development and be present in the environment as adults. Adult fly collections are often ignored in cases of advanced decomposition because of a presumed little relevance to the investigation; herein we present information on how these insects can be of value. In this study we applied an age-grading technique to estimate the age of adults of Chrysomya megacephala (Fabricius), Cochliomyia macellaria (Fabricius), and Phormia regina (Meigen), based on the temperature-dependent accumulation of pteridines in the compound eyes, when reared at temperatures ranging from 5 to 35°C. Age could be estimated for all species*sex*rearing temperature combinations (mean r2±SE: 0.90±0.01) for all but P. regina reared at 5.4°C. These models can be used to increase the precision of post-CI estimates for remains found indoors, and the high r2 values of 22 of the 24 regression equations indicates that this is a valid method for estimating the age of adult blow flies at temperatures ≥15°C.

Decomposition of Concealed and Exposed Porcine Remains in the North Carolina Piedmont.

We examined the decomposition and subsequent insect colonization of small pig carrion (Sus scrofa (L.)) placed in concealed and open environments during spring, summer, and fall in Raleigh, North Carolina, as a model for juvenile human remains. Remains were concealed in simulated attics in three manners, ranging from minimal to well-concealed. Concealment had a significant effect on the insect community colonizing the remains across all three seasons; the beetles Necrobia rufipes (DeGeer) (Cleridae) and Dermestes maculatus (DeGeer) (Coleoptera: Dermestidae) were the only species indicative of remains located indoors, whereas numerous fly (Diptera: Calliphoridae, Muscidae, Sepsidae, and Piophilidae) and beetle (Coleoptera: Silphidae, Staphylinidae, and Histeridae) species and an ant species (Hymenoptera: Formicidae, Prenolepis sp.) were indicative of remains located outdoors. Season also significantly affected the insect species, particularly the blow flies (Diptera: Calliphoridae) colonizing remains: Lucilia illustris (Meigen) was indicative of the spring, Cochliomyia macellaria (F.) and Chrysomya megacephala (F.) were indicative of the summer, and Calliphora vicina Robineau-Desvoidy and Calliphora vomitoria (L.) were indicative of the fall. In addition, across all seasons, colonization was delayed by 35­768 h, depending on the degree of concealment. These differences among the insect communities across seasons and concealment treatments, and the effects of concealment on colonization indicate that such information is important and should to be considered when analyzing entomological evidence for criminal investigations.

Persistence and Retention of Porcine Reproductive and Respiratory Syndrome Virus in Stable Flies (Diptera: Muscidae).

We investigated the acquisition of porcine reproductive and respiratory syndrome (PRRS) virus by the stable fly (Diptera: Muscidae; Stomoxys calcitrans (L.)) through a bloodmeal, and virus persistence in the digestive organs of the fly using virus isolation and quantitative reverse-transcription PCR (qRT-PCR). Stable flies were fed blood containing live virus, modified live vaccine virus, chemically inactivated virus, or no virus. Stable flies acquired PRRSV from the bloodmeal and the amount of virus in the flies declined with time, indicating virus did not replicate in fly digestive tissues. Virus RNA was recovered from the flies fed live virus up to 24 h postfeeding using virus isolation techniques and 96 h using qRT-PCR. We further examined the fate of PRRSV in the hemolymph of the flies following intrathoracic injection to bypass the midgut barrier. PRRSV was detected in intrathoracically inoculated adult stable flies for 10 d using qRT-PCR. In contrast to what we observed in the digestive tract, detectable virus quantities in the intrathoracically inoculated stable flies followed an exponential decay curve. The amount of virus decreased fourfold in the first 3 d and remained stable thereafter, up to 10 d.

Development of a novel walk-through fly trap for the control of horn flies and other pests on pastured dairy cows.

A prototype walk-through fly vacuum system, designed to remove horn flies Haematobia irritans (L.) (Diptera: Muscidae) from cattle, was developed and tested for efficacy. The study was conducted during 4 fly seasons over 17 consecutive weeks each year within the months of May through September at 1 dairy research herd in the coastal plain of North Carolina. Additional data on horn flies, as well as face flies (Musca autumnalis) and stable flies (Stomoxys calcitrans), were collected during 1 yr from 7 commercial pasture-based and organic dairy farms in the piedmont region of North Carolina. The number of flies observed on animals in the pasture was compared with the number of flies collected in the trap. Studies were initiated after horn fly densities had met or exceeded a threshold of 200 flies per animal. The vacuum trap removed between 1.3 and 2.5 million flies annually from the research station cattle. Most fly removal occurred during the first few weeks of operation and maintained densities below threshold thereafter. Cattle using the fly trap at the research farm had only about 28% the number of horn flies as untreated cattle, and reductions ranged from 67.5 to 74.5% across the 4-yr study. In addition to large numbers of horn flies, traps placed on commercial dairies during 1 yr collected stable flies, face flies, and house flies, all species with differing behavior and larger in size than horn flies. The estimated cost of running the trap is $72 per season at commercial rates of $0.12 per hour and an expected 4h of daily operation during the time of milking. Use of a vacuum system as described herein has potential as a cost-effective method in reducing populations of parasitic flies in pasture-based dairy production systems without the use of insecticides.

Molecular epidemiology of Staphylococcus aureus mastitis in dairy heifers.

The specific purpose was to investigate the possible interrelationships of genotypes of Staphylococcus aureus found in mammary glands, horn flies, and extramammary sites on 3 southeastern US dairies. A total of 1,228 samples were obtained from various sources on the 3 dairy herds, each of which had a history of Staph. aureus mastitis. Dairy herds studied had access to pasture, and samples were collected during the summer when horn flies (Haematobia irritans) were active. Samples collected included milk samples from all lactating herd cows, colostrum samples from heifers calving during the study period, heifer body sites (mouth, nostrils, and teats), the heifer environment (water, feed, and soil/vegetation/pasture), horn flies, and humans (hands and nostrils). Isolation of Staph. aureus was attempted from all samples, with isolates subjected to genotypic analysis using pulsed-field gel electrophoresis. A total of 244/1228 (or 19.9%) of all samples were positive for Staph. aureus. For milk samples, 52/383 (or 13.6%) of samples were Staph. aureus positive, and 70/411 (or 17.0%) of heifer quarter colostrum samples were positive. Horn fly samples were frequently positive, with over one-half (29/52, or 55.8%) of samples positive for Staph. aureus. Staphylococcus aureus obtained during the study comprised isolates from 12 different genotype groups as defined in this study. Identical genotypes were obtained from horn flies, heifer colostrum samples, and cow milk samples. Group B genotypes were shared among flies, heifer colostrum samples, body sites, and cow milk samples, whereas group A genotypes were common to the same sample locations and body sites but rarely (once) found in horn flies. We conclude, based upon the finding of identical pulsed-field gel electrophoresis genotypes in flies, heifer body sites, and heifer colostrum samples, that flies and heifer body sites could be important sources of Staph. aureus for heifer intramammary infections.

Blow flies (Diptera: Calliphoridae) survive burial: Evidence of ascending vertical dispersal.

This study was undertaken to determine if immature blow flies could complete development following burial and emerge from the soil as adults. Two species of blow flies, Cochliomyia macellaria and Protophormia terraenovae, were placed at three depths and at three different life stages, in a simulated burial to evaluate the impact of soil on ascending vertical dispersal and fly survival. In soil columns, immature stages of each species were covered with 5, 25 and 50cm of soil. Emerging adult flies of both species reached the surface from all depths at all three immature stages (2nd instar, 3rd instar and pupae). At the 50-cm depth, flies were least successful in reaching the surface when buried as pupae and most successful as late 3rd instar larvae (prepupae). Collectively, more adult flies emerged from the soil if buried as 3rd instars (79.6%) than either 2nd instars or pupae (59.6% and 59.3%, respectively (F(2,159)=14.76, P<0.0001)). Similarly, at shallow burial depths of 5 and 25cm, 75.6% and 70.4% of the adults successfully reached the surface, compared with 52.6% at the 50-cm depth (F(2,159)=15.95, P<0.0001). Second instars demonstrated ascending vertical dispersal behaviours in the soil column by pupating closer to the surface. Nearly half (46.6%) of the C. macellaria 2nd instars buried in 25cm of soil pupated nearer to the surface. Similarly, 45.4% of the P. terraenovae 2nd instars pupated nearer to the surface. When buried at 50cm, approximately 25% of 2nd instars of both species pupated nearer to the surface. When 3rd instars of C. macellaria and P. terraenovae were buried at 120cm, 40% and 4.3% of the adults, respectively, successfully reached the soil surface.

Assessment of Stomoxys calcitrans (Diptera: Muscidae) as a vector of porcine reproductive and respiratory syndrome virus.

Porcine Reproductive and respiratory syndrome (PRRS) is a globally significant swine disease caused by an arterivirus. The virus replicates in alveolar macrophages of infected pigs, resulting in pneumonia in growing pigs and late-term abortions in sows. Outbreaks occur on disparate farms within an area despite biosecurity measures, suggesting mechanical transport by arthropods. We investigated the vector potential of stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), in the transmission of porcine reproductive and respiratory syndrome virus (family Arteriviridae, genus Arterivirus, PRRSV) under laboratory conditions. Stable flies were collected around PRRS-negative boar stud barns in North Carolina and tested for presence of the virus. Stable flies were collected on alsynite traps placed near the exhaust fan of the close-sided tunnel-ventilated buildings, suggesting blood seeking flies are attracted by olfactory cues. No flies were positive for PRRSV. We assessed transmission of the virus through an infective bite by feeding laboratory reared stable flies on blood containing virus and transferring them to naive pigs for subsequent bloodmeals. Transmission of the virus to naive pigs by infective bites failed in all attempts. The volume of blood contained within the closed mouthparts of the stable fly seems to be insufficient to deliver an infective dose of the virus. Stable flies are unlikely to transmit PRRSV from one pig to another while blood feeding. The fate of the virus after a bloodmeal remains to be determined.

Novel field assays and the comparative repellency of BioUD(®) , DEET and permethrin against Amblyomma americanum.

Two new field bioassay methods were developed to compare the repellent activity of BioUD(®) (containing 7.75% 2-undecanone), 98.1% DEET and 0.5% permethrin against natural populations of nymphal Amblyomma americanum (L.) (Acari: Ixodidae). In a cloth sheet assay, pieces of material measuring 41 × 58 cm, separately treated with one of the test materials or the appropriate solvent carrier, were placed at random on the ground and baited with dry ice for 1 h. Mean numbers of ticks on repellent-treated sheets were significantly lower than on control sheets. There was no significant difference in the number of ticks collected between sheets treated with BioUD(®) and those treated with DEET. However, significantly fewer ticks were found on sheets treated with BioUD(®) or DEET than on permethrin-treated sheets. In a sock test, over-the-calf tube socks were treated with one of the test materials or the appropriate solvent carrier. Human volunteers wore a repellent-treated and a corresponding carrier-treated sock on either leg and walked randomly over an area of approximately 4000 m(2) for 15 min. Significantly fewer ticks were collected from socks treated with BioUD(®) or DEET than from socks treated with the carrier and there was no significant difference in repellency between these two agents. No difference in the mean number of ticks collected was found between permethrin-treated and corresponding carrier-treated socks. To examine the mechanism of repellency of BioUD(®) , a four-choice olfactometer was used to assess spatial repellency against adult A. americanum. As expected in the absence of a repellent, when all choices were represented by water-treated filter paper, ticks were equally distributed among the choices. When one choice consisted of BioUD(®) -treated filter paper and the remaining choices of water-treated paper, the distribution of ticks on the repellent-treated paper was significantly lower than might be expected to occur by chance, suggesting that repellency is at least partly achieved by an olfactory mechanism.

Efficacy of the new repellent BioUD against three species of ixodid ticks.

BioUD with the active ingredient 2-undecanone originally derived from wild tomato plants is a new repellent recently registered by the US EPA. Repellent efficacy of BioUD (7.75% 2-undecanone) and DEET (98.11%) was examined in the laboratory using a choice test between repellent-treated and control filter paper surfaces for Amblyomma americanum, Dermacentor variabilis, and Ixodes scapularis. BioUD provided greater repellency against A. americanum and I. scapularis than DEET. No difference was found between BioUD and DEET against D. variabilis. In head-to-head assays between BioUD and DEET, undiluted and 50% dilutions of BioUD were more repellent than undiluted DEET against all three species tested. Similarly, a 25% dilution of BioUD was more repellent than DEET against A. americanum while no difference in mean percentage repellency was found between a 25% dilution of BioUD and DEET against I. scapularis. Based on regression analysis, the concentration of BioUD required for equivalent repellency to 98.11% DEET was 39.5% for D. variabilis and 29.7% for I. scapularis. A log-probit model could not be constructed for A. americanum from the dosages tested. Based on filter paper head-to-head assays, BioUD is at least 2-4 times more active as a repellent than DEET against three species of ixodid ticks under the conditions of our laboratory bioassays.

Managing the horn fly (Diptera: Muscidae) using an electric walk-through fly trap.

An electric walk-through fly trap was evaluated for the management of the horn fly, Hematobia irritans (L.), on dairy cattle in North Carolina over 2 yr. The trap relies on black lights and electrocution grids to attract and kill flies that are brushed from the cattle passing through. During the first season, horn fly densities were reduced from >1,400 to <200 flies per animal. Horn fly density averaged 269.2 +/- 25.8 on cattle using the walk-through fly trap twice daily, and 400.2 +/- 43.5 on the control group during the first year. The second year, seasonal mean horn fly density was 177.3 +/- 10.8 on cattle using the walk-through fly trap compared with 321.1 +/- 15.8 on the control group. No insecticides were used to control horn flies during this 2-yr study.

Vector competence of Musca domestica (Diptera: Muscidae) for Yersinia pseudotuberculosis.

The vectoR potential of adult house flies. Musca domestica L., for Yersinia pseudotuberculosis (Pfeiffer), a pathogen of domestic animals and humans, was investigated. Adult flies were allowed to feed on trypticase soy broth (TSB) containing Y. pseudotuberculosis for 6 h and then transferred to sterile containers with sterile TSB as a source of water and nutrients. At 6-h intervals, all flies were transferred to sterile containers with sterile TSB and 10 randomly selected flies were examined for the pathogen. Yersinia pseudotuberculosis did not establish a permanent population in the house fly colony; however, viable cells were detected from the digestive tract of flies for up to 36 h after the initial exposure, and flies contaminated their environment (sterile TSB) for up to 30 h after the exposure. These results demonstrated that house flies can carry Y. psedotuberculosis for a considerable period and therefore must be considered as a potential mechanical vector of pseudotuberculosis infection.

Diversity and contribution of the intestinal bacterial community to the development of Musca domestica (Diptera: Muscidae) larvae.

The bacterial diversity in the intestinal tract of Musca domestica L. was examined in larvae collected from turkey bedding and corn silage. Aerobic culturing yielded 25 bacterial species, including 11 from larvae collected from turkey bedding and 14 from larvae collected from corn silage. Providencia rettgeri (Hadley, Elkins & Caldwell) was the only species common to both environments. Two mammalian pathogens, Yersinia pseudotuberculosis (Pfeiffer) and Ochrobactrum anthropi (Holmes), were isolated from the larval intestinal tracts. The majority of isolates represented facultatively anaerobic heterotrophs capable of fermentation. The significance of these bacteria for development of house fly larvae was evaluated by bioassays on trypticase soy egg yolk agar. Pure cultures of individual bacterial species isolated from the intestinal tract of larvae from turkey bedding supported development of flies to a much greater extent than those isolated from larvae from corn silage. House fly development was best supported by a Streptococcus sanguis (White) isolate. The significance of bacteria for development of house flies is discussed.

Limited transmission of turkey coronavirus in young turkeys by adult Alphitobius diaperinus (Coleoptera: Tenebrionidae).

We examined the role of lesser mealworm, Alphitobius diaperinus (Panzer), in the transmission of an enteric disease of turkeys caused by a coronavirus. Turkey coronavirus (TCV) from two sources was studied, one isolate (NC95) was embryo propagated, the second was TCV infected material from turkeys diagnosed with poult enteritis mortality syndrome (PEMS). Beetles were fed virus-infected feces mixed with chicken feed. Transmission of virus was effectively halted by surface sterilization of the beetles. Turkey poults administered beetle homogenates infected with TCV+ PEMS that had not been surface sterilized had reduced weight gains and 50% mortality. Mortality and weight gains were not effected in the NC95 group. Virus isolation procedures were performed to determine NC95 viability at varying time intervals. Beetles were dissected and the guts removed 1, 12, and 24 h after the initial viral feeding. Whole beetles were also examined for comparison. Whole beetles and beetle guts were homogenized and injected into turkey eggs for embryo propagation. Direct immunofluorescence was used to determine the presence of TCV. A. diaperinus were capable of mechanical transmission of TCV. However, only turkey embryos receiving whole beetle and beetle gut homogenates within 1 h of feeding on the virus were positive for TCV. Laboratory studies demonstrating PEMS transmission by A. diaperinus are continuing.

Fungal mastoiditis in immunocompromised children.

The immunocompromised host is subject to a variety of opportunistic infections. Mycotic infections, including invasive fungal sinusitis, are a dreaded complication in immune deficient children. Fungal mastoiditis has rarely been described in this population. Our experience with 2 cases of fungal mastoiditis in immunocompromised children is reviewed. Case histories describing aggressive medical management with and without surgical intervention and a review of the literature are presented.

Attachment-site patterns of adult blacklegged ticks (Acari: Ixodidae) on white-tailed deer and horses.

The attachment site pattern of adult Ixodes scapularis Say on white-tailed deer and horses in Maryland was determined by whole-body examinations during fall and spring periods of tick host-seeking activity. On deer in the fall, both female and male I. scapularis attached largely to anterior dorsal body regions, with attachment to the ears (outside), head, neck, and brisket accounting for 87.9% of females and 86.6% of males. The attachment pattern of females differed between bucks and does during fall, but not in spring, and both females and males were more abundant on bucks than does during fall, but not in spring. Neither female nor male attachment patterns on deer differed between fall and spring seasons. In contrast to deer, the ears and neck of horses were largely devoid of blacklegged ticks, and 84% of the females were attached either on the chest, in the axillae of the fore and rear legs, or under the jawbone. The restricted attachment of female blacklegged ticks to ventral body regions of horses may reflect avoidance of light. An understanding of the attachment patterns of adult I. scapularis, an increasingly abundant and economically important species, enhances sampling of feeding ticks, deticking to limit host irritation or exposure to tick-borne pathogens, and identifies body areas that should be targeted for delivery of repellents or acaricides.

Density and distribution of cattle lice (Phthiraptera:Haematopinidae, Linognathidae, Trichodectidae) on six steers.

The density and distribution of four species of cattle louse, Bovicola bovis (L.). Haematopinus eurysternus (Nitzsch), Linognathus vituli (L.), and Solenopotes capillatus (Enderlein), were elucidated from the hides of six slaughtered steers. Adult and nymphal lice were first removed from one hide by hand and the location of each specimen mapped. The remaining lice were removed by a detergent wash, and KOH dissolution of hide and hair. Lice from the remaining five hides were removed using KOH dissolution of cattle hair and subsequent filtration of the effluent. Bovicola bovis was most abundant, followed by H. eurysternus, L. vituli and S. capillatus. Significant variation was observed in B. bovis, H. eurysternus and L. vituli population densities. Solenopotes capillatus population densities did not differ significantly. All species were contagiously distributed, i.e. 'clumped', suggesting species dependent predilection sites. Predilection sites were ranked according to louse density to facilitate the development of field sampling strategies. Additional biological data were gathered on sex and life stage ratios for each species.

Egg yolk and bacteria growth medium for Musca domestica (Diptera: Muscidae).

Six preparations of egg yolk media inoculated with Escherichia coli (Migula) were evaluated as an in vitro growth system for immature house flies, Musca domestica L. Larval development, based on pupation and eclosion success, was normal on all media supplemented with E. coli, but few larvae survived without bacteria (4%). Pupation (72%) and eclosion (63%) were significantly higher on egg yolk media without mannitol than on media with mannitol or on blood agar controls.

Evaluation of field propagation of Muscidifurax zaraptor (Hymenoptera: Pteromalidae) for control of flies associated with confined beef cattle.

The parasitic wasp Muscidifurax zaraptor Kogan & Legner was mass-reared in the field to control house flies, Musca domestica L., on two Nebraska beef cattle confinements. About 50,000 freeze-killed house fly pupae were exposed to a single release of M. zaraptor in the field. Placement of six additional cohorts of 50,000 freeze-killed pupae at the release sites at 2-wk intervals resulted in a mean parasite emergence of 56.4% over the study period. Mean fly mortality of 37.3 and 25.9% occurred in sentinel pupae placed around the perimeter of two release sites, compared with 3.9% for two control sites. We demonstrated a negative correlation between host reduction in sentinel cohorts and distances the cohorts were placed from parasite release sites. However, data indicated that other environmental factors also influenced the success of M. zaraptor in locating sentinel hosts. Correlation between mortality in sentinel pupae and numbers of parasites released was not evident. Temperatures above approximately 28 degrees C appeared to reduce the effectiveness of M. zaraptor.