Overview of the USDA-ARS Center for Grain and Health Research and Stored Product Research Unit in Manhattan, Kansas

James E. Throne

U.S. Department of Agriculture, Agricultural Research Service, Center for Grain and Animal Health Research , 1515 College Avenue, Manhattan, KS, USA Center for Grain and Animal Health Research

Agricultural Research Service CGAHR has five research units - four located in main facility on 12 acre (4 hectare) site and one unit on Kansas State University campus

Main facility

50,000 bu (700 mt) Research Grain Elevator Engineering and Wind Erosion Research Unit

 4 scientists  Develop technologies for rapid, automated assessment of grain quality – detecting disease, mycotoxins, or insect infestation; quantifying protein and other quality traits  Develop techniques for safe storage of grain – identity preservation, dust reduction  Control wind erosion of the soil – Wind Erosion Prediction System Grain Quality & Structure Research Unit

 5 scientists  Determine grain characteristics and components responsible for end-use quality – e.g., which proteins in wheat affect bread rising  Find new uses for sorghum  6 scientists  Develop disease- and insect-resistant wheat that can tolerate environmental stresses

Leaf Rust

Hard Winter Wheat Genetics Research Unit

Head Scab

Karnal bunt

Hessian fly -Borne Animal Diseases Research Unit

 7 scientists  Conduct research to solve major endemic, emerging, and exotic arthropod-borne disease problems in U.S. livestock Stored Product Insect Research Unit

 7 scientists  Develop environmentally friendly and cost-effective methods to control stored-product insect pests Which stored products do we work with?

• Bulk grains in bins and elevators

• Milling, processing, and warehouse industry

• Transportation industry - railcars, ships

• Urban environments - stores, homes Why work on stored-product ?

• Insects reduce the quality of stored grain and other stored products in the U.S. and throughout the world. • Over 10 billion bushels (254 million metric tons) of corn, 2 billion bushels (54 million metric tons) of wheat, and about a billion bushels (27 million metric tons) of barley, oats, rice, rye, and sorghum are grown in the U.S. each year, with a value of over 44 billion dollars. • It is estimated that postharvest losses to these grains due to insects are 5 to 10% in the U.S., or about 2.2 to 4.4 billion dollars per year. • Losses in developing countries are estimated at 30 to 50%. • Losses to processed commodities are difficult to quantify, but probably greatly exceed the losses to raw commodities. • Many of the insecticides used by the cereal foods industry are being lost due to insecticide resistance or regulatory changes, so we need alternative control technologies. Dr. Frank Arthur • Integrated Pest Management

He conducts studies to determine optimal use of insecticides, particularly reduced-risk insecticides and aerosols, to help industry optimize insecticide use in stored grain and in processing facilities. He investigates effects of various factors such as temperature, relative humidity, and sanitation on efficacy of insecticides. Dr. Jeff Lord • Insect pathology

Many of the reduced-risk 400 350 30oC control technologies have 300 lower efficacy than 250 conventional insecticides, 200 150 so we are looking for ways 100 to synergize insect 50 pathogens to make them 0 Check Bb DE 75 more effective, such as Bb+DE 43 combining the fungus Beauveria bassiana with diatomaceous earth. Dr. Paul Flinn • Expert Systems

The expert system Stored Grain Advisor was developed to aid in pest management in farm-stored wheat. Stored Grain Advisor Pro was developed to predict risk of insect problems in grain stored at elevators.

Dr. Paul Flinn • Biological control

These small parasitic wasps attack beetle larvae in stored grain. He found that by reducing the grain temperature using aeration in the summer, these beneficial insects were 10 times more effective. Dr. Paul Flinn • Biological control

Matt Grieshop was a graduate student studying the behavior and ecology of tiny parasitoid wasps that are used to suppress moth pests in retail environments. Dr. Jim Throne • Ecology and sampling

We conducted a study that showed that the current grain buying practice based on insect-damaged kernels (IDK) is not a reliable indicator of insect infestation in railcars of wheat because there is not a clear relationship between IDK and the insect infestation level present in the grain. Dr. Jim Throne • Ecology and sampling

Entomological applications of near-infrared spectroscopy developed include detection of insect-infested kernels and quantification of insect fragments in flour. Dr. Jim Throne • Ecology and sampling

Insects developing inside kernels can be detected using digital X-ray technology. X-rays provide the most accurate estimate of internal insect infestation, but the method is laborious and expensive

Digital x-ray images obtained with Faxitron MX-20

Lesser grain borers in rice

Digital x-ray images obtained with Faxitron MX-20

Rice weevils in rice

Digital x-ray images obtained with Faxitron MX-20

Lesser grain borers in wheat

Digital x-rays are very rapid, but equipment is expensive and can scan only a 10 X 10 cm (4 X 4 inch) area at a time – ca. 5 grams of wheat or ca. 175 kernels Electrically Conductive Mill Electrically Conductive Mill - Commercial version processes 2 kg/min Electrically Conductive Mill

Insect stage Detection rate

2nd or 3rd instar 74%

4th instar or 83% pupae

No false positives Electrically Conductive Mill

• High detection rate for large larvae and pupae and 0% false positive rate • Only works for live insects because it measures moisture; this could be a problem for grain that has been fumigated and destined for milling • Able to inspect 2 kg in a minute with no sample preparation • Commercially available Dr. Jim Throne • Ecology

Liposcelis entomophila

Current studies are on ecology % of original number of nymphs surviving to adults and control of psocids, an 100 emerging pest of stored grain and processing, warehouse, 80 and retail facilities, such as 60 determining efficacy of heat, aerosols, and crack and 40 % surviving to adults % surviving crevice treatments for control 20 or determining optimal 0 temperature and grain type Control Methoprene Carrier EsfenvalerateCombined for development. Aerosol treatments Most psocid pests of stored products are from the genus - 7 species of Liposcelis and that are pests of stored products have been reported from the U.S.; they don’t have common names and they are difficult to identify to species - , Liposcelis bostrychophila, Liposcelis brunnea, Liposcelis corrodens, Liposcelis decolor, Liposcelis entomophila, and Liposcelis paeta - The first two species are parthenogenetic 2005 Psocid trends – 115,059 psocids collected 100

10 Grain samples

Bin 1 - 248 total psocids Bin 2 - 299 total psocids 1 10000

1000

100 Surface Refuges

10 Bin 1 - 14,829 total psocids Bin 2 - 18,786 total psocids

1 1000

100 Hatch Refuges 10 Bin 1 - 1,427 total psocids Bin 2 - 1,968 total psocids 1 10000 log(Number ofpsocids log(Number 1)+

1000 Manual Insector Counts 100 Bin 1 - 45,284 total psocids 10 Bin 2 - 32,218 total psocids

10000

1000 Electronic Insector Counts

100 Bin 1 - 29,902 total psocids Bin 2 - 35,153 total psocids

10 Sep Oct Nov Dec Jan Feb Mar We determined how long it takes to kill adult psocids at different temperatures

95% FL Temperature Lepinotus reticulatus Liposcelis entomophila (hours) (oC) at:

LT95 37.5 90.92 (93.00-102.35) 111.43 (100.19-126.88) 40.0 13.83 (12.59-15.56) 43.52 (39.90-48.23) 42.5 3.84 (3.50-4.36) 9.06 (6.96-15.68) 45.0 0.70 (0.67-0.74) 5.51 (5.16-5.95) 47.5 0.66 (0.63-0.70) 1.89 (1.81-1.99) Psocids appear to be very susceptible to heat, particularly Lepinotus reticulatus, but they are known to leave a facility to escape fumigants Chlorfenapyr and β-cyfluthrin provided efficient control of Liposcelis bostrychophila and L. entomophila at the label rates, unlike pyrethrin

Insecticide Liposcelis LT95 species (95% FL) [hours] β-Cyfluthrin entomophila 12.5 (11.7-13.6) bostrychophila 15.3 (14.6-16.2) Chlorfenapyr entomophila 5.1 (4.9-5.4) bostrychophila 7.7 (6.9-9.1) Pyrethrin entomophila 102.1 (94.6-112.5) bostrychophila 195.8 (158.3-280.1)

Chlorfenapyr is derived from a class of microbially-produced compounds known as halogenated pyrroles; β-Cyfluthrin is a synthetic pyrethroid Efficacy of sulfuryl fluoride for control of stored-product psocids

100 100

80 80

60 60

40 40 adults nymphs 20 20

0 0 0 1 1.5 2 4 6 8 10 12 24 48 96 0 1 1.5 2 4 6 8 10 12 24 48 96

mortality (%) 100

80 Mean mortality (% ± SE) of L. paeta adults, nymphs, and eggs 60 after 48 h of exposure to SF at 40 eggs various doses 20

0 0 1 1.5 2 4 6 8 10 12 24 48 96 SF dose (g/m3) Dose to 100% kill of psocids after 48 h of exposure to SF Dose (g/m3) Egg Nymph Adult Lepinotus 24 4 8 reticulatus Liposcelis - - 6 bostrychophila Liposcelis 72 - 24 decolor Liposcelis - 6 4 entomophila Liposcelis paeta 96 6 6 Dr. Jim Campbell • Insect Behavior

Movement of self-marked warehouse beetle males in a food facility shows for the first time how insects can move from one part of a facility to another to reinfest after treatments, such as fumigation. Dr. Jim Campbell • Insect Behavior

Pest populations are being monitored in flour mills to determine the impact of fumigation treatments Fumigation on the populations, and the dates relationships between inside and outside populations with pheromone trap capture and product infestation. Dr. Brenda Oppert • Insect proteomics

We are looking at protein profiles of insects and how to exploit those proteins for insect control. Dr. Dick Beeman • Insect genomics

CHITIN SYNTHASE 1 CHITIN SYNTHASE 2 LG5

marker cM 22.P3t 40.1 12E1131.G5t 43.90.0 The red flour beetle is the PIG124 44.5 32.H2s 44.5 35.B1s 45.2 first agricultural pest to 14A05 46.0 36.I1s 46.0 20C02 46.0 31.L5s 46.0 have its genome sequenced 36.D10s 46.8 32.O1scs1 46.8 28.B16t 46.8 as a result of a collaboration 19.C3s 48.1 13G03 49.8 3F12 49.8 31.M1t 50.4 between SPIRU, KSU, and 32.H16s 51.7 12G02 51.7 24.B1t 51.7 the Baylor College of 16B10 51.7 27.P24t 51.7 C17A08 52.8 12C07 53.4 Medicine. We anticipate Txn049 54.0 PU123 54.0 that mining the genome will A81-2 54.0 lead to totally new ways to control insect pests. Chitinase gene knockout by RNA inhibition (RNAi)

Sixteen chitinase genes have been found via genomics. The figure shows results of inhibition of just one of these genes. The other 15 chitinases are still functional, yet the insect can’t molt and dies because it can’t digest the old cuticle, which therefore remains thick and stiff so the insect becomes entrapped when it tries to shed this old cuticle. normal one chitinase knocked out Cuticle cross-linking gene knocked out by RNAi

normal yellow-e gene vacuum-pack knocked out de-aeration of food There are 14 yellow genes involved in cuticle tanning and hardening. The cuticle-crosslinking gene yellow-e is required for waterproofing of the cuticle. Knockout of yellow-e results in loss of the waterproofing function of the exoskeleton and in rapid desiccation and death. The insect becomes “vacuum-packed” in its own skin. DNA fingerprinting for population/infestation analysis

individual #1 A B B B A C

individual DNA fingerprints #2 of six beetles from the same warehouse, showing 3 different genotypes, A, B & C

Individual B differs from A at 2 positions out of 23 in this DNA segment. There are 6 million other segments of this length in the genome. to applied research SPIRU covers all areas of stored-product entomological research:

from basic research Get further information and download publications at http://ars.usda.gov/npa/gmprc/spiru