Duration: 10/01/2001 to 09/30/2006

Administrative Advisor(s):

NIFA Reps:

Non-Technical Summary

Statement of Issues and Justification

Synanthropic arthropods, particularly flies and stored product insects, are characteristically associated with human activity. Although these insects are stimulated to enter the home, they are frequently associated with animal agriculture because they live and breed in the manure, spoiled feeds and other organic materials on the farm. In an effort to meet the needs of a growing human population animal agriculture has changed significantly from the small family farm to intensive and sometimes vertically integrated systems (Axtell 1999). As these farming systems increased in size and number of animals, so did quantities of manure and other insect breeding materials. Management of manure and the insects associated with manure is a challenge in poultry and livestock systems.

Insects are rarely included in manure and waste management projects. Often insects are rare, for example; processing large quantities of animal waste using anaerobic lagoon digestion systems or advanced technologies like extrusion and pelleting of manure solids do not promote insect breeding. Unfortunately the associated risks from the environmental impact of lagoons to the high costs of extrusion and pelleting restrict the broad use of such waste processing systems. Other methods such as dry pit storage, deep stacking, open floor production on dry bedding, dry lots, and composting are more feasible for many growers, but these create substantial accumulations of soiled bedding, litter, manure or compost.

Under such circumstances pest management is critical before, during and after storage and subsequent to land application of litter, manure or compost. Recognized as serious economic pests, flies and litter beetles are extremely difficult to control (Harrington et al. 1999). Fly and beetle nuisance complaints often result in litigation, impacting both the farm and the farming community (Hunter 1997). Legal cases concerning nuisance flies and beetles often occur following the land application of animal waste. Rich in nitrogen (N), phosphorous (P), and potassium (K), animal manure is often applied to field soils as an organic fertilizer. Over use of organic fertilizers has resulted in state and federal regulation through animal waste management plans. The fate of N, P, and K of manure applied to field soils is a concern in concentrated production areas as there may not be enough cropland within economical hauling distance to properly use the manure. The development alternative systems that reduce manure nutrients through management, and reduce the insects associated with animal wastes will lead to better efficient utilization of these materials.

In addition to flies and other manure-breeding insects, animal wastes contain known foodborne pathogens. The CDC reports over 300,000 individuals are hospitalized annually and 76 million endure foodborne illnesses. Food related illnesses are associated with a variety of foods, particularly contaminated meat, poultry, egg products, and organic produce (CDC 1997). Escherichia coli, Campylobacter jejuni, Listeria monocytogenes, and Salmonella spp are among the leading foodborne bacterial pathogens in the US (Mead et al. 1999). The role of filth breeding insects in the dissemination and transmission of these bacteria is important to the development of pest management strategies intended to limit the spread of bacteria within the farm, and between farms and the community. Furthermore, manure management may directly affect pathogens in the manure and the safety of subsequently produced organic vegetables.

Clearly manure and pest management is essential to reducing nuisance issues in the community.

Understanding the role of these insects in the dissemination of foodborne pathogens may provide insight into the management of these bacteria in the pre-harvest interval. In this project we propose to evaluate manure management relative to filth flies, litter beetles and other insects. Manure management systems vary with commodity and region. We have selected manure management plans relative to geographic region. For example: caged-layer production in NY state relies on close-sided high-rise poultry houses with deep stacked manure, while in NC open-sided houses with wood shaving litter prevail for the meat-bird market. Each member of this multistate project will concentrate on manure management practices within their respective states. Each has investigated nuisance fly complaints and made recommendations based on solving the immediate problem. In this study we concentrate on developing a standardized method of monitoring nuisance insects and developing methods to reduce pest densities before nuisance thresholds are met. Since many nuisance complaints arise following the land application of organic fertilizers, we will also investigate the impact of manure and litter incorporation relative to soil type and condition for the management of nuisance insects. Lastly, our efforts will concentrate on the role on filth breeding insects in the harborage and dissemination of foodborne pathogens, studying the dynamics and seasonality of the relationship between insect and pathogen, and the relative importance within the farm community and the pre-harvest interval.

Stakeholder Needs: Recent surveys of poultry producers established the relative importance of filth flies and beetles associated with poultry. Flies were problematic for 22% of the NC meat bird industry (Toth et al. Unpublished data). Flies (56%) were reported as a primary pest of caged-layer production in New York poultry facilities (Harrington et al. 1999). Over 65% of NY producers felt that flies were resistant to insecticides. Fly control costs to the poultry industry were estimated at $32.4 million annually (Thomas and Skoda 1993).

North Carolina poultry producers ranked the darkling beetle their number one pest (Toth et al. Unpublished data). Darkling beetles were reported by 41% of NY poultry producers as causing economic loss (Harrington et al. 1999). Annual losses to darkling beetles have been estimated at $16 million in Virginia and $10 million in Georgia. These loss estimates are generally attributed to energy costs in beetle-damaged broiler houses, and replacement of beetle-damaged insulation. To date, no economic data exists on beetle vectored disease losses because we know little of the prevalence of disease organisms.

Management of these pests on the farm relieves livestock, poultry and employees from persistent annoyance associated with their habits and behavior. With the suburbanization of rural communities, the threat of litigation is ever present (Thomas and Skoda 1993). Recently litigants in Ohio were awarded millions of dollars in judgements against poultry producers (Miller 1997, Olejnik 2001). As a result, producers are often willing participants in applied research projects, offering use of facilities and labor. Producer interaction enables us to better develop cost-effective strategies that will work in their systems. Because of their continued involvement and assistance, producers readily adopt innovation and continue to support research and extension programs, allowing their farms to serve as applied laboratories.

Pest Species

House flies, Musca domestica, feed on a wide range of organic matter including manure and mixtures of manure, bodily excretions and decaying organic matter. The house fly is an important pest in poultry production. Dispersing distances of 2.3 to 11.8 km in 24 hours, public nuisance issues are a growing concern (Greenberg 1973, Thomas and Skoda 1993). House flies constitute a health hazard as well as an annoyance. Habitually these insects are likely to pick up disease-causing organisms and contaminate human food by crawling on the surface and depositing feces and regurgitated liquid. House flies have been implicated in the spread of over 30 bacterial and protozoan diseases.

The darkling beetle, Alphitobius diaperinus (a.k.a. litter beetle) has emerged as an important arthropod pest of poultry production (Axtell 1999). The beetle is a pest in two life-stages, mature larvae and adults. Mature larvae climb up the walls and posts of poultry facilities and chew into wood support structures and insulation weakening the structure and increasing energy consumption (Vaughn et al. 1984, Despins et al. 1987). The adult stage is a pest when manure is spread on fields. Evidence presented in litigation (Miller 1997) suggests that beetles move en masse toward artificial lights generated by residences near fields on which beetle-infested manure has been spread. The mechanism inducing such migration is poorly understood but important to solving nuisance and disease issues relative to this pest. The litter beetles is a known reservoir of wide variety of avian (and human) pathogens and parasites, including Salmonella typhimurium, Escherichia coli, tapeworms, avian leucosis virus, and turkey enterovirus (Axtell and Arends 1990, Despins et al. 1994, McAllister et al. 1994, 1996).

Poultry Production Systems

Each of the three poultry facility types principally used in poultry production in the U.S. (caged-layer, broiler, and breeder houses) has unique pest management needs. Caged-layer houses are widely used for commercial egg production and present the greatest fly and darkling beetle breeding potential. Broiler houses are wide-span structures with litter (wood shavings) covering the floor and the birds running free. Little fly breeding occurs because of the dry litter; but high populations of beetles may occur in the litter. Breeder or broiler-breeder houses are also wide-span structures with birds running free on a slat-litter floor. The outer two-thirds of the house has a slatted floor 2 to 3 feet above ground level, with a litter-covered floor in the center third of the house.

Related, Current and Previous Work

Results of a CRIS search indicate 373 projects involved with foodborne pathogens. The focus of these projects varied from detection methodology, and environmental contamination, to post-harvest contamination of foods. Few CRIS projects concentrated on the pre-harvest interval with an emphasis on host-pathogen interactions. Two projects include tritrophic interactions between pathogen, host and vectors; Steelman, ARK-01819 and Watson, NC-06503 include insects in the CRIS description relative to foodborne pathogens. Both are members of this multistate project. CRIS search results indicate 127 projects working on nuisance issues related to manure management focusing was on odors and environmental issues. Project MIN-17-050, Moon, concentrates on the biology and management of muscoid flies, including nuisance fly monitoring. Moon is a participant in the current project. Two terminated projects were listed in the CRIS search, Pitts, PEN-03294 (terminated in 1998) concentrated on integrated management of manure odors relative to nuisance fly issues and Sheppard, GEO-00172, (terminated in 2000) developed much of the black soldier fly biology applied in the current new-project description.

National and regional priorities relate to the disposal of manure. Our project is critical because many manure management CRIS projects emphasize environmental concerns but fail to address entomological issues. The climatic, animal husbandry and environmental variation between regions influence the potential of pest insects to become a nuisance. In fact, regional variation affects the means by which the pest population may be managed. For example the proposed treatment, Black Soldier Fly manure digestion, has been proven to control house fly populations in the southern states but its application in northern regions has not been explored.

The proposed work is consistent with national and regional priorities of producing viable alternatives to traditional waste management systems, reducing pathogens in our food supply and promoting a better relationship between agriculture and an expanding non-farm public. Regional differences in poultry production require regional collaborative efforts of all the participants. Veterinary entomologists from universities, state experiment stations, ARS and CSREES are represented in this project. This coalition is positioned to address problems in poultry production systems within to their respective regions; Southeastern meat bird production, deep South and Northeastern caged layer operations or the expanding Midwestern poultry industry. Our goals are to reduce foodborne pathogens and related nuisance insect concerns relative to regional manure management options. With a growing human population within these respective regions, this project fosters a better relationship between animal agriculture and the community at large.

Likely impacts of successfully completing the work are that poultry producers will have the knowledge and tools needed to manage flies, improve biosecurity, reduce nuisance outbreaks and limit the spread of foodborne pathogens among poultry products. This will mean that producers will be able to increase bird growth rates, and egg production. These increases in productivity will exceed control costs, and increase the profit margins for these producers. Producers will be able to reduce fly and beetle numbers thereby reducing conflicts with neighbors, accurately identify fly nuisance levels should they occur and have knowledge of the risk and reduce the incidence of food-borne illness. The benefits offered through this project will provide for a better relationship between producers and residents at the rural-urban interface and provide for a safer food supply.

Conventional Manure Handling:

Deep stack manure is currently the caged-layer industry standard. In meat-bird production, free birds are placed on a 10-15 cm layer of litter (wood shavings). Depending on condition, litter may be used for successive flocks before removal and land application. Higher manure and litter moisture promotes insects.

Composting

Composting manure involves thermophilic processing and decomposition by aerobic microorganisms to produce a relatively stable organic material. High temperature significantly reduces pathogen and insect survival. There is considerable interest in composting, but few operations compost manure. Manure alone is not conducive to composting because of offensive odor and poor physical properties. Combining manure with a high-carbon bulking agent improves aeration and the physical properties of the manure.

Effects of composting on fly production potential have been examined in two recent studies. Pitts et al. (1998) tilled manure beds under caged laying hens and found that composting increased bed temperatures above 50 0C, decreased moisture content below 40%, and virtually eliminated larval house flies. Moon et al. (2001) studied the effects of composting on the subsequent nutritional value of poultry manure for larval house flies. Composting in a windrow system reduced the manure=s fly production potential by 90%.

Black Soldier Fly

The black soldier fly (BSF) is a southern native, non-pest fly that unlike the house fly, is not attracted to human habitation or foods (Furman et al. 1959). BSF reduce manure accumulations 42-56% and give 94-100% house fly control through larval competition and by repelling ovipositing house flies (Bradley and Sheppard 1984). Elimination of lesser mealworm has been noted, but not well documented. The digested residue is a friable compost-like material with about 24% less nitrogen (net loss of 60%). BSF is being investigated as a possible feedstuff for swine, poultry, and several species of fish (Sheppard and Newton 2000).

Land Application

As part of a routine manure management plan, land application is a practical means of disposing large amounts of animal wastes and an efficient use of an organic fertilizer. Unfortunately manure may contain a large number of house fly larvae and pupae, and beetles. Under S-274, Hogsette (1996) reported that fly larvae successfully complete development in sand with >1% manure solids added. Mechanical incorporation of manure is often recommended to help reduce odor and was thought to reduce the potential for a fly or beetle outbreak. Watson et al. (1998) evaluated house fly survival following the incorporation of manure into a gravel loam soil by disk, harrow and moldboard plow. Adult flies reached outbreak proportions 10 days following application, regardless of treatment, and the outbreak continued 11 days. In contrast darkling beetle survival appears to be impacted by incorporation of poultry litter into piedmont red clay soils (Watson et al. 2001). Soil type may have an impact on the successful use of this cultural practice for pest control. Other manure management methodologies may also reduce insects or pathogens before the material is land applied.

Monitoring

Since the above methods are considered industry standards for manure handling, it is imperative that we develop accurate and sensitive house fly and beetle sampling methods. Thus standardizing the evaluation methods for measuring the pest impact in agricultural and non-agricultural areas. Currently, veterinary entomologists and public health officials rely on techniques suitable on the farm. For example, Lysyk and Axtell (1986) evaluated several methods of monitoring adult fly population in poultry houses including the speck card, which has become the standard for monitoring adult flies for confined livestock and poultry. Fly densities are estimated by counting the specks on standard index cards and taking an average of several cards to establish a mean. Although adequate for determining fly densities, no information on speciation is available. Similarly, Hogsette et al. (1993) used sticky cards to monitor fly densities during a 24 hr period. The number and species of fly on the cards were counted to estimate the fly densities. Scudder grills were developed to monitor fly densities in military kitchens and mess halls. Although used routinely since 1944, the Scudder grill has not been fully validated (Scudder 1996). Mark and recapture studies conducted in Minnesota (O=Rourke 2001) evaluated baited jug traps, white sticky cylinders and a modified Scudder grill. Trap types were equally sensitive to changes in fly abundance, but the jug traps were far more efficient at catching flies when actual abundance was moderate to low. Performance of these traps and experimental alternatives remains to be evaluated in other regions of the U.S.

Litter beetles can be sampled in poultry facilities using three methods: (1) visual estimation, (2) direct manure sampling and (3) trapping. Visual sampling for litter beetles is inherently difficult and often inaccurate because of the cryptic nature of the beetle. The manure core method provides an accurate estimate of beetle populations; however, the method is labor intensive and not applicable to outdoor environments. The current standard is the tube trap (Safrit and Axtell 1984), in which corrugated cardboard is rolled and placed inside a cylinder of PVC pipe. The trap is placed on the manure or litter surface and emptied weekly, providing a measurement of changes in beetle abundance over time. Unfortunately, the traps are relatively ineffective at low to moderate beetle densities and the use of the trap outdoors is untested. One method for improving trap design to measure low densities would be to incorporate attractants into the traps.

Pathology

Food-borne pathogens originating in manure interact with insects in various ways. A potentially beneficial interaction occurs when flies of certain species reduce the pathogenic bacteria. For example, Morgan (1995) reported that medicinal maggots (Calliphoridae) reduce bacterial infections when used in wound therapy. In the case of BSF digestion of manure as a management process, potential food-borne pathogens may be reduced. Because the black soldier fly is one of our treatments, we will be examining BSF for the presence of foodborne pathogens.

The most commonly considered negative interaction occurs when adult insects vector these pathogens to humans or food animals (Greenberg 1973, Kobayashi et al. 1999). The adult house fly, and to a lesser extent, the adult darkling beetle, is of primary concern as these insects have highly developed dispersal abilities and pose the greatest risk of pathogen transmission (Despins et al. 1994, McAllister et al. 1996). Flies and beetles can be sampled to monitor the presence and prevalence of selected foodborne pathogens on the farm and in the outlying community. For this new project we will gather information on the prevalence and seasonality of foodborne pathogens in house flies and darkling beetles.

Pest Management

The management of these pests has bearing on the prevalence of these organisms in the community. IPM strategies include cultural, biological, and chemical methods to control the targeted pests. A portion of this project uses and develops cultural and biological control strategies to reduce pests. In addition to manure treatments and black soldier fly many other agents may be included, parasitoids, predators and entomopathogenic fungi (Axtell 1999).

Insecticides are important for controlling a pest outbreak, yet there are few insecticide options available to support livestock and poultry IPM. House flies are resistant to permethrin and tetrachlorvinphos and increasingly resistant to cyfluthrin (Scott et al. 2000, Kaufman et al. 2001). Four residual premise materials are registered for use against darkling beetles, tetrachlorvinphos (organophosphate), carbaryl (carbamate) lambda-cyhalothrin and cyfluthrin (both pyrethroids). Resistance is suspect in carbaryl, and two chemicals, lambda-cyhalothrin and cyfluthrin can only be used in vacant buildings, and tetrachlorvinphos is currently under FQPA review.

Objectives

1. Evaluate conventional and experimental poultry manure management systems as they influence production of filth flies, litter beetles and associated foodborne pathogens.
   
2. Evaluate novel cultural, biological and chemical strategies for pest management to minimize nuisance and health risk in the rural-urban interface.
   
3. 
   

Methods

Objective 1: Evaluate conventional and experimental poultry manure management systems as they influence production of filth flies, litter beetles and associated foodborne pathogens.

I. Manure Management Systems: Treatments (NC, NY, IN, AR, GA, TN)

Environmental concern stemming from over use of land applied manure demonstrates a need to reduce manure volume and nutrients (nitrogen), insect pests, and pathogens before land application. However in some areas, manure is directly applied to the land unaltered. Within this objective, we will evaluate the insects and pathogens commonly associated with poultry manure as impacted by manure handling systems, including BSF digestion, composting and tarping.

Standardized sampling will be used for all manure treatments. Manure core samples will be collected for fly larvae and beetle enumeration. All manure core samples will be placed into Tullgren funnels where live insects will be extracted and counted. Insects collected from the different manure treatment systems will be examined for the presence of bacteria. Methods are outlined in Appendix I. Further analysis of nutrient value of each treatment will be conducted including N, P, K, and trace elements and heavy metals for all treatments.

IA. Conventional Manure/Litter Systems (GA, NC, IN, NY, FL-ARS)

Currently industry standards for handling manure include deep stack (caged layer) in NY, IN, FL, GA, MN, MI and litter (meat birds) in NC, AR, MN. These systems will serve as the standard to which all other manure-handling systems are compared across regions. Methods used to sample this system for insect and pathogens will be dependent upon those used in subsequent treatments.

IB. Black Soldier Fly Digestion (GA, NC)

A black soldier fly population will be developed at a 96 hen experimental caged layer house in Tifton, GA. Natural oviposition will be augmented with young larvae from our BSF colony if necessary (Tomberlin and Sheppard 2001). Deep stack manure and associated data will be collected from a nearby commercial caged layer house. Each treatment will be sampled monthly for house fly larvae, litter beetles and pathogens associated with these and the manure. Aquaculture trials determining the value of Black Soldier Fly as a feed stuff for talapia and catfish will be conducted in GA and NC. Expanded application of the poultry based BSF system into the NC swine industry is being investigated. High-rise hog and retro-fit belt systems for hog houses are under development.

IC. Composting (AR, IN, NY, NC)

IC.1. In-house composting. (NY) Caged-layer poultry farms are increasingly utilizing in-house composting as a manure management strategy (Pitts et al. 1999). Using this system, composted manure must be removed every 6 to 12 weeks allowing multiple evaluations per season. Manure will be sampled weekly for presence of arthropods. Furthermore, each week additional samples will be characterized for fly growth and development, moisture will be added to samples using an appropriate serial dilution to produce a range of moisture levels and live house fly eggs will be introduced (Moon et al. 2001). Fly growth and development will be recorded. This information will demonstrate the importance of complete vs. partial compost.

IC.2. Outdoor Composting. An evaluation of outdoor composting will be conducted. In these systems the producer moves manure out of the facility every two days. We will sample manure for house flies and darkling beetles monthly during the time period that beetles are active. Furthermore, additional compost samples will be removed to the laboratory and evaluated for house fly breeding potential. (MN, NY, MI).

IC.3. Remove and Cover. Manure from caged-layer poultry facilities in NY and FL infested with darkling beetles will be removed from buildings and a minimum of five piles (manure heaped until piles are 2 m long by 1.5 m wide) will be formed outdoors (smaller version of standard producer practices). Data loggers will record manure and ambient temperatures hourly. All piles will then be covered with a tarp and sealed. Manure core samples will be taken 1, 3, 7, 14 and, if necessary, 21 days after tarping. Manure will be applied to a nearby field after day 21. This process will be repeated four times annually for a minimum of two years. Full-size manure piles on farms that utilize this management strategy will also be sampled and monitored using the same protocol.

Broiler or turkey litter infested with darkling beetles will be sampled and enumerated from facilities in NC as described previously. Litter from these facilities will be used to evaluate three methods of litter treatment, 1) roof covered pile, 2) piled and tarped (as above) and 3) bagged and composted. In covered pile treatments, litter is stored in building until spread. Bagged composed treatments are designed to place litter into large plastic bags currently designed to hold silage (i.e. Ag-Bags) but modified to provide air necessary for composting. Manure will be left in these bags for up to one month. Post-bag sampling will be conducted as described previously.

II. Outdoor Management of Nuisance Insects

IIA. Determine Standard monitoring methods for flies, beetles and other related pests of confined livestock and poultry, (MN, NC, IN, AR, GA).

IIA.1. Suitability of commercial and experimental fly traps for monitoring abundance of house flies and blow flies will be examined through comparative studies. Baited jug traps, white sticky cylinders and baited pyramid traps will be compared by placing replicates of each design at 10-m intervals in proximity to known sources of house flies. The traps will then be rotated among positions over consecutive days according to a Latin square design. Relative efficiency will be evaluated by comparing mean daily catch rates of males and females of each species of fly. Using these methods poultry operations with a history of nuisance fly outbreaks will be targeted (NC, NY). To address the dichotomy between on farm thresholds and nuisance fly thresholds the fly population densities will be monitored inside and outside the facility. Resident nuisance threshold will be established and correlated to the fly densities within the facility. Working with the farm, an IPM program will be formed to reduce fly nuisance complaints (see Objective 2).

IIA.2. Darkling beetle migration. (NC) Anecdotal evidence suggests that darkling beetles respond to lights and poultry odors. A minimum of 25,000 adult darkling beetles will be starved for 3 days then given feed labeled with Nile Blue or Sudan Red dyes. The beetles will be allowed to consume labeled feed for 7-14 days to stain the cuticle and fat body. Aged turkey litter, free of darkling beetles, will be applied to the field surface a minimum of 1000 meters distance from the turkey house. Litter will not be incorporated into the field soils. The marked beetles will then be taken to the field and released.

Sticky cylinder traps will be placed around the house at 10 meter intervals to capture flying beetles. Sticky ribbons fixed to the threshold of all openings to the turkey house. Tube traps (Safrit and Axtell 1984) will be place around the outside of the building to capture walking beetles. Battery powered sticky light traps will be placed at 100 meter intervals around the field perimeter. Collected beetles will be frozen until evaluated for the presence of stain.

IIB. Evaluate survival of insects following incorporation according to region. (NC, IN, NY, MI)

Poultry manure/litter containing all life stages of house flies or darkling beetles will be collected from caged-layer (NY) and broiler houses (NC) and loaded into a manure spreader. The litter will be spread onto approximately 500 square feet of field according to the current state recommendations for litter/manure application based on nitrogen content. The soil and soil conditions will be characterized. Treatments will be assigned to the 10 by 50-ft rows and replicated 4-5 times. Treatments will include: control (no incorporation), mulch till (15 cm), disk (7.5 cm), and moldboard plow (30 cm). Following incorporation, emergence traps of either stove-pipe cylinders for beetles or cone traps for flies will be used to monitor survival. Square tiles and pitfall traps are well suited for beetle sampling. Ten tile and pitfall traps per treatment row (minimum of 30 traps/treatment) will be placed randomly in the field. Alsynite sticky traps will be placed on a conduit pole 4 ft above ground to sample flying insects. We propose 16 treatment rows, 32 traps at the row ends plus 3 traps on each side of the field for a total of 38 alsynite traps. All traps will be examined on day 8, 14, 18, 21 and 25 where beetles will be removed and counted from each trap.

III. Food safety and health issues of manure and insects relative to manure treatments

The aforementioned treatments of manure and litter are suspected to alter the microflora within treatments. It is important to record changes in selected foodborne pathogens that may be present before and after treatment and in any associated insects. In this section we will concentrate on determining the impact of treatment on four common pathogens and the role of insect in the dissemination of foodborne bacteria. (NC, MI, TN, FL, GA)

IIIA. Pathogens Monitored

Four pathogens are responsible for much of the foodborne illnesses in the US. These include Camplyobacter, Listeria, Salmonella species and E. coli. Methods for the isolation and identification of these bacteria range from relative simple culture media to biochemical tests and PCR. We have outlined the basic methodology for isolation and identification of bacteria from insects and manure/litter samples in the appendices.

IIIB. Evaluate the relationship between E. coli, Salmonella spp., Listeria monocytogenes, and Campylobacter jejuni and muscoid fly prevalence.

IIIB.1. To establish the presence of foodborne bacteria, adult flies will be collected from five broiler farms and five turkey farms (10 total) monthly using sweep-net or sticky cards. Freeze killed flies will be surface sterilized and the crops aseptically removed. Cultures of bacteria will be identified according to the methods previously described.

IIIB.2. Monitor the movement and dispersal potential of muscoid flies in the transmission of selected strains of bacteria within and between farms. Flies will be captured at distances from putative sources to determine the possibility for flies to transmit and disperse bacteria. Characteristic bacterial profiles of flies collected from the farms will be compared with profiles of flies collected from the adjacent community.

IIIB.3. Monitoring of the density dependent relationship of flies to bacterial prevalence in adjacent poultry houses on 10 farms (a): The relationship of fly, bird, and bacteria will be directly compared using bacterial characterizations described above. Muscoid fly densities will be monitored using spot cards and sticky cards. Fecal samples (cloacal swabs) will be taken from 10 birds randomly selected from the flock.

IIIB.4. The ability of adult house flies to acquire, harbor and excrete (oral and fecal) E. coli from artificially inoculated manure will be investigated. Methods developed under S-274 will be modified for house flies (McAllister et al.1996). In these studies, house fly adults will be placed on manure with known added quantities of E. coli and monitored for feeding activity. An additional experiment will examine the length of time that adult house flies retain internally. Each day, flies will be placed on a sterilized agar plate and allowed to vomit and defecate. These plates will be held to allow for bacterial growth and determination of E. coli presence.

IIIB.5. Seasonal prevalence of foodborne bacteria within poultry houses and dairies

The role of the house fly in the transmission of foodborne bacteria is limited by the seasonal abundance of pathogens and the flies themselves. During this monitoring period we will obtain data on the seasonal abundance, prevalence and transmission potential of flies for these important bacteria. Using the aforementioned methodology we will monitor flies and fly breeding substrate(s) and the presence of characterized bacteria throughout the study.

Objective 2: Evaluate novel cultural, biological and chemical strategies for pest management to minimize nuisance and health risk in the rural-urban interface. (IN, NC, NY, MN, GA)

The need for new pest management strategies in livestock and poultry pest management has never been greater. Although several insecticides are currently available, the implementation of the Food Quality Protection Act, FQPA, will likely remove a number of these materials. In all likelihood, we will be an industry with few options as re-registration progresses. There is a critical need to evaluate new chemistries and actively search out unregistered products that hold promise to work against flies and beetles. The acceptance of biological control by producers is now widespread. We have an understanding of these processes within limited geographical areas, but not on a multi-regional basis. Furthermore, the addition of new biological control organisms holds the promise of increasing our biological arsenal. Finally, it is critical that we understand the resistance status of flies and beetles to our currently registered materials.

I. Adulticides

Efficacy studies of new products against targeted pests are essential. Unfortunately many new chemistries come to livestock and poultry well after its use in other commodities. For example, pyriproxifen, has been evaluated for activity against many row crop pest and only recently the house fly and darkling beetle. Inclusion of this insecticide in IPM programs requires further study of the non-target effects as well. We will examine pyridine in replicated laboratory studies and determine the impact of the insecticide on second and subsequent parasitoid generations. We will also examine the material against immature C. pumilio (NC, NY, IN, TN, MN, FL, GA).

II. Biological Control

Research on prospective new classical biological control agents of house flies will be conducted by UMN, and ARS-FL. Lines of Eurasian pteromalid wasps that were recently obtained from Russia and Kazakhstan (under S-274) will be compared to counterparts already present in the US. The Eurasian lines appear to be Muscidifurax raptor, Spalangia endius, S. cameroni and S. nigroaenea. Reproductive isolation among Eurasian and North American lines will be evaluated directly by reciprocal crosses (MN), and indirectly by comparing mitochondrial and nuclear DNA sequences. Temperature requirements, reproductive capacity, searching abilities and host ranges of the different lines will be evaluated in the laboratory using standard methods (MN, ARS-FL). Performance of lines that appear to be promising will be tested against house flies under field conditions.

III. Insecticide Resistance

Assays will be developed that utilize a serial dilution of technical-based insecticides applied to filter papers (Sheppard and Hinkle 1987). Adult house flies will be assayed for susceptibility to permethrin, cyfluthrin, tetrachlorvinphos, dimethoate and methomyl. Both adult and larval beetles will be assayed for susceptibility to tetrachlorvinphos, carbaryl and cyfluthrin, the only registered materials. House fly and darkling beetle resistance status will be determined for a minimum of 5 farms (poultry) in several states (NY, NC, GA, FL, IN, MN). Determination of darkling beetles to instar is difficult. Larvae will sieve sized to obtain mid to late instar specimens. These will serve as representatives in the larval resistance assay. An additional group of larvae will be held in the laboratory for adult emergence. Darkling beetle adults are fairly long lived (3B12 months), therefore, we will test beetles 1-3 weeks after adult emergence.

Measurement of Progress and Results

Outputs

• We will determine the impact of various manure handling systems on the presence of house flies, darkling beetles and several pathogens. Our results will establish the best manure management approach within each region, and make knowledgeable recommendations to producers. We will develop standard monitoring methods for flies and beetles that will allow for an unbiased and accurate determination of nuisance evaluations. Results will determine the impact of manure incorporation on a multi-regional scale such that producers may avoid community conflicts. Our final outcome under this objective will be a critical understanding of the temporal and spatial distribution and role of the house fly in the transmission of several potentially dangerous foodborne pathogens.</P>
• We will develop new pest management strategies that can be cost-effectively implemented by producers. The results obtained in this critical objective will enable producers to avoid confrontation with their neighbors by eliminating the source of the confrontation. Furthermore, a collaborative, multi-regional, standardized resistance-monitoring program will be established that will allow for the implementation of effective resistance management strategies.

Outcomes or Projected Impacts

• Under Objective 1, we anticipate the manure treatments will provide alternatives to conventional manure management practices. Black soldier fly digested and composted manure have value as soil amendments through reduction in available nutrients in finished product lessening environmental impacts. These treatments are expected to reduce or eliminate foodborne pathogens that could enter the foodchain following spreading of manure and litter resulting in a safer fertilizer for organic growers. We anticipate manure inhabiting arthropods to be involved in the dissemination of foodborne pathogens on the farm and perhaps between farm and outlying communities. Pest control and manure treatment recommendations developed in this study will be important in the management and control of foodborne illness. </P>
• Standardized monitoring methods for flies, beetles and other related pests of livestock and poultry are essential to establishing origin for nuisance evaluation. This will allow producers to evaluate ambient densities of manure breeding insects in their immediate environments, and will provide them and neighboring citizens with tools necessary to resolve nuisance complaints.
• Preliminary results from NC suggest that incorporation of manure into piedmont clay soils results in reduced dispersal from fields (Appendix Figure 1). We suspect that similar results will be obtained in the heavy Midwestern and Northeastern soils, but are uncertain as to the results in the sandy coastal soils in NC and GA. </P>
• It is expected that house flies will be resistant to permethrin, probably tetrachlorvinphos and less so to cyfluthrin in poultry facilities. Regional differences are expected to emerge with resistance greatest in the southern region. Darkling beetle resistance is unknown. Understanding the resistance status of these insects will allow producers to more appropriately manage these important pests.
• Although new insecticide chemistry is slow to emerge, novel compounds used in other commodities will be evaluated as to their effectiveness against pests and impacts on non-target insects under Objective 2. Our studies will yield new, effective pest management strategies including biological control. Recent collections of pteromalid parasitoids from Eastern Europe may provide new options for more efficient biological control programs. Furthermore, recent advances with currently used pteromalids have provided a better understanding of the optimum combinations and release strategies in specific regions. These advances will be evaluated in multiple regions providing valuable data.

Milestones

(2002): Year 1 Insecticide resistance profiles of house flies and darkling beetles will be established for each cooperating region (NC, FL, GA, TN, MI, MN, AR, IN, NY). Manure and litter treatments established by region and nutrient analysis initiated (NC, FL, GA, TN, MI, MN, AR, IN, NY). Begin pathogen and insect evaluations (NC, MI, GA, TN, FL, AR). Establish contacts with farms with histories of nuisance complaints and begin testing devices for outdoor nuisance pest evaluations, initiate monitoring (MN, NC, NY).

(2003): Year 2 Continue manure and litter treatments. Begin supplemental reductions of insects prior to incorporation experiments. Conduct incorporation experiments. Continue nuisance pest monitoring (MN, NC, NY). Begin seasonality and prevalence monitoring of selected bacteria in the house fly and darkling beetle (NC, MI, GA, TN, AR).

(2004): Year 3 Continue manure and litter treatments. Practicality of manure treatment on a regional basis established. Continue supplemental insect reduction and incorporation experiments relative to outdoor nuisance pest evaluations. Correlate nuisance and on farm fly thresholds. Seasonality and prevalence of selected bacteria in the house fly and darkling beetle will be established. Development of outreach, demonstration and education programs.

(2005): Year 4 Impact of pest management on the prevalence of selected bacteria established. Outdoor nuisance pest devices defined and evaluated. Demonstration projects underway to reduce on farm fly densities relative to nuisance thresholds and dissemination of foodborne bacteria.

(2006): Year 5 Final year of demonstrations of best management practices. Insecticide resistance profiles conducted to monitor change from yr. 1.

(0):0

Projected Participation

View Appendix E: Participation

Outreach Plan

The project Website will be managed by the Center for IPM, North Carolina State University, under the management of Dr. Ron Stinner. Similar to the current S-274 website (http://cipmtest.ent.ncsu.edu/s274/), this site will link to participant relevant websites.

We will generate several fact sheets as appropriate and add updates to the Pest Management Recommendations of each state. Our materials are currently made available to many states extension services and posted regionally, e.g., the Cornell University Veterinary Entomology and NY State IPM WWW site. Each regional site contains extensive extension materials and links to related web sites.

Results of this research will be made available to producers through a number of venues. Annual poultry conferences are routinely hosted by project participant, AR, IN, MN, NY, NC, GA, and FL, for example, New York/New England Poultry Pest Management Conference. Poultry pest management information relevant to Minnesota, North Carolina and Florida follow similar venues, including independent and contract growers, industry representatives, company integrators, public health officials and others. Similarly, University of Minnesota and North Carolina State University and Purdue University will host workshops for poultry producers in their respective states. Results of the present research will be incorporated into those workshops as presenters, planners and producers deem to be appropriate.

Organization/Governance

Chair: Presides over the annual meeting and prepares the annual report for submission to the Adminstrative Representative.

Chair Elect: Nominations are taken from the membership. The chair elect is chosen from a body of candidates voted upon by the membership at the annual meeting.

Secretary: the Past Chair assumes the position of the Secretary. The Secretary is responsible for writing the minutes of the annual meeting and having the minutes posted on the Website.

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Attachments

Land Grant Participating States/Institutions

AR, CA, FL, GA, IN, KS, MN, NC, NY, TN

Non Land Grant Participating States/Institutions

USDA-ARS/Florida