Topics in Sustainable Ecological Agriculture

 

Microbial control of grasshoppers, under changing climate and management.
General summary of background and progress.

 

Key funding collaborators:
Alberta Crop Industry Development Fund;
Saskatchewan Pulse Growers;
Pesticide Risk Reduction Program, Agriculture and Agri-Food Canada Pest Management Centre;
National Defence, Cold Lake 4 Wing (biocontrol in Environmental Programs).

and program funding from
Canada Foundation for Innovation
Canada Research Chairs
NSERC

Project staff at the University of Lethbridge:

PI: Dan Johnson
Student research assistants in 2007: Toni Pasolli, Jeff Moker, Samuel Wind, Josh Phillips, Jon Siminovitch, and Alia Adatia
Student research assistants in 2008: Eric Macknak, Patrick Chan
Research Technician in 2009: Patrick Chan.
Student research assistants in 2009: Brittany Turcotte, David Zhang, Craig Wiebe, Lena Arjannikova
Previous Research Staff: Susan Entz, MSc.
Student research assistants in 2010: Lena Arjannikova, Alexis Kaminski, Craig Wiebe, David Zhang, Qiuying Zhang and Sichen Zhou .
Additional volunteers: Adil Adatia, Eric Johnson, Sam Johnson, Lucy Martin-Johnson, Meggy Martin-Johnson, Christine Wilson, and Jim Yu.
Environmental Staff of Cold Lake 4 Wing.

Thanks also to Mark Goodwin, Pulse Canada.

Related International collaborations: USDA, and China.

Address:

Dan L. Johnson
Professor of Environmental Science
BSc: USask (Biology, High Hon.);
MSc, PhD: UBC, Dept of Plant Science,
Institute of Animal Resource Ecology.

Department of Geography (Biogeography)
University of Lethbridge, 4401 University Drive W.
Lethbridge, Alberta, Canada T1K 3M4.

Office WESB1020: (403) 329-2040, Students (403) 332-4047
Lab WESB1020 (403) 317-5056, Fax: (403) 332-4039.

2010-2012 research projects; papers in preparation or submitted:
- environmental safety of Metarhizium for aquatic arthropods (3 species, separate experiments)
- environmental safety of Metarhizium for rainbow trout
- environmental safety of Metarhizium for earthworms
- temperature extremes and growth of Metarhizium - formulation and production of Metarhizium for insect control
- effects of adjuvants and insecticide mixtures on Metarhizium germination, survival and growth
- effects of weather on field activity of biopesticides

July, 2010. dan.johnson@uleth.ca

The initial work toward alternatives was related to a need for tools and plans for management of anticipated increased numbers of grasshoppers under warm conditions (part of Climate Change Impacts and Adaptation Program, CCIAP; Project A526: "Prediction of expected impacts of climate change on grasshopper abundance and species composition in the Canadian Prairies".). We have also investigated the probable impacts of weather on the potential of biological control of grasshoppers with Metarhizium. Current research is directed at refinement of production, formulation, ecological interactions, environmental sustainability, field effectiveness against key target species, and the effects of environmental conditions.

Biopesticide development summary poster.

Johnson., D., Goodwin, M., Irvine, P., Kaminski, D., Byrne, J., Johnson, E., Hartley, S, and Olson, B. 2010. Sustainable management tools for growers: a made-in-Canada biopesticide option, combined with an early warning of risk, and accurate pest identification. Poster Presentation, Pulse Days 2010 (Jan 11-12, 2010, Saskatoon, SK).

 

Basic photos in Powerpoint form (PDF file)

For operational use, licensed under government regulations, spores can be produced on grain (barley and related grains are used in North America; rice is used in Africa and Asia). Spores are combined with oil and sprayed for control of grasshoppers. The method is routine and widely used with a range of microbial control agents. An early, possibly the first, case of the use of fungal spores in oil for grasshopper control in the field was:

Johnson, D.L., Goettel, M.S., Bradley, C., Bradley, van der Paauw, H., and Maiga, B. 1992. Field tests of the entomopathogenic fungus Beauveria bassiana against grasshoppers in Mali, West Africa, July, 1990. In: C. Lomer & C. Prior (eds). Biological Control of Locusts and Grasshoppers, CAB International, Wallingford, UK, pp. 296-310

(French edition: Johnson, D.L., Goettel, M.S., Bradley, C., van der Paauw, H., et Maiga, B. 1992. Essais en plein champ du champignon entomopathogéne Beauveria bassiana contre les criquets au Mali, en juillet 1990. Dans: Lutte biologique contre les acridiens, préparé par C. Lomer et C. Prior, pp. 298-313. CAB International, Wallingford, UK, et CIDA/ACDI, Hull, Canada)

Replicated field tests in 2008 were very successful, with significant reductions in target insect density after 6 days, and over 80% reduction 15 days after treatment. Field tests in 2009 were less successful because they were conducted in dry pastures and grass cover with lower height and biomass, near drying cereal crops. This biopesticide is not highly effective in stopping highly mobile insects that move into a crop days after treatment application.

Spores used in field applications, 2008-2009

Metarhizium anisopliae S54, isolated from Alberta soil, is produced on grain for research testing. The final spore product has low moisture content (6.7% moisture by weight, determined by weighing before and after drying at 60 C), produced in small-scale production experiments in May-June, 2009). Yields are typically 40-60 g per kg grain. The final dry spore product has slightly more than 4 X 10^10 spores per g. Below: Metarhizium anisopliae S54 spores produced on plates and grain at the University of Lethbridge, in 2009
Metarhizium anisopliae S54 colonies are coin-sized and have high production. Repeated serial re-isolation of some entomopathogens can result in declining virulence. (Aug 8, 09, DJ)	Conidia form in densely pack structures
Fresh spores from test-scale lab production, used in field experiments in 2009	Dry spores added to water with wettable oil separate well (used in field-testing in 2009)
Spores in a rejected batch, with contaminants visible	A perfect, 100% pure batch of spores, from April 27, 2010

 

Spores dispersed in the oil-surfactant-water emulsion, in the lab	Spores recovered from a drop of spray tank solution
Scanning electron micrograph of dry conidia (Metarhizium anisopliae S54)	Chains of spores from a colony on a PDA plate
Metarhizium anisopliae S54 (on dodine plates, April 27, 2010, DJ)	colonies of Metarhizium anisopliae S54 (on dodine plate)

 

July, 2010. dan.johnson@uleth.ca

 

Quality assurance during research production, isolation and application. Quality assurance under future industrial production would follow established methods already in use by the manufacturers of other isolates of Metarhizium. Quality assurance in a laboratory setting is simpler because of the smaller quantities, but more challenging because of the need to conduct experiments concerning optional substrates, nutrient additives, source of contaminants, possible correction of contaminated cultures, and factors affecting entomopathogen virulence and viability during storage and field application.
Contamination from air or from substrate material is a problem with any low-cost method of fungal production. But with the use of even a basic microscope, recognition and rejection of test materials containing contaminants is not difficult.
If you can tell these apart, ...   then you can just as easily recognize the major unwanted microbes.
Alternaria is common in indoor and outdoor air. Plates exposed in office buildings commonly yield it.	Several common species of Aspergillus are yellow to green, depending on growth stage. The cleistothecia stages (fruiting body) are round and flower-like.
Aspergillus spores are round and easy to distinguish from Metarhizium.
Other common lab and household contaminants include Rhizopus, Mucor and bacteria. Contamination should be avoided and not allowed to produce unwanted growth, which must be controlled by disinfectants and autoclaving (normally 121 C) of substrates and containers.

 

 

Environmental safety experiments have been encouraging. So far, experiments have concerned the following non-pest organisms:

Birds: ring-necked pheasants tested with related isolates, but so far not with the Alberta isolate

Fish: planned for 2009, in collaboration with the Aquaculture Centre of Excellence, Lethbridge College. New tests with fresh spores planned.

Earthworms: in progress, May, 2009; Replicated trials planned for 2010.

 

Other invertebrates:

1. Aquatic (common in ponds and streams on the Canadian Prairies)

a. amphipod

Gammarus
Common name: scud (adult and sub-adult)
Class Crustacea, Order Amphipoda, Family Gammaridae

b. aquatic insects

Chaoborus americanus (Johannsen)
Common name: phantom midge (free-swimming larva)
Class Insecta, Order Diptera, Family Chaoboridae

Coptotomus longulus LeConte
Common name: small predaceous diving beetle (adult)
Class Insecta, Order Coleoptera, Family Dytiscidae

Notonecta undulata Say
Common name: backswimmer (adult)
Class Insecta, Order Hemiptera, Family Notonectidae.

2. Terrestrial

a. beetles (Coleoptera)

Amara littoralis Mannerheim
Common name: ground beetle (adult)
Class Insecta, Order Coleoptera, Family Carabidae
(Pterostichus melanarius, common black ground beetle, was not available)

Coccinella septempunctata (Linnaeua)
Common name: seven-spot ladybird, or 7-spotted lady beetle (adult)
Class Insecta, Order Coleoptera, Family Coccinellidae

Epicauta pennsylvanica (De Geer)
Common name: black blister beetle (adult)
Class Insecta, Order Coleoptera, Family Meloidae

Harpalus funerarius Csiki
Common name: ground beetle (adult)
Class Insecta, Order Coleoptera, Family Carabidae

Tenebrio molitor Linnaeus
Common name: yellow mealworm, or yellow mealworm beetle (adult and larva; article submitted)
Class Insecta, Order Coleoptera, Family Tenebrionidae.

c. grasshoppers and katydids (Orthoptera)

i. Pest species (tested to confirm infectivity against target species. The main test species were

ii. Non-pest orthopterans tested

Chorthippus curtipennis (Harris)
Common name: marsh meadow grasshopper (adult)
Class Insecta, Order Orthoptera, Family Acrididae

Conocephalus saltans (Scudder)
Common name: prairie meadow katydid (adult)
Class Insecta, Order Orthoptera, Family Tettigoniidae

Orchelimum gladiator Bruner
Common name: gladiator katydid (adult)
Class Insecta, Order Orthoptera, Family Tettigoniidae

Pseudopomala brachyptera (Scudder)
Common name: bunchgrass grasshopper, or toothpick grasshopper (adult)
Class Insecta, Order Orthoptera, Family Acrididae

d. parasitic wasps (Hymenoptera)

Trichomalopsis sarcophagae (Gahan)
Common name: fly parasitoid wasp (adult; in collaboration with Dr. Qi Hu and Dr. K. Floate)
Class Insecta, Order Hymenoptera, Family Pteromalidae

e. Soil microarthropds

Folsomia candida Willem
Common name: springtail (adults and juveniles; small lab tests)
Class (or Subphylum) Hexapoda, Order Collembola, Family Isotomidae

Pest species:

Ceutorhynchus obstrictus (Marsham)
Common name: cabbage seedpod weevil (adult; small lab tests)
Class Insecta, Order Coleoptera, Family Curculionidae

Phyllotreta cruciferae Goeze
Common name: crucifer flea beetles (adult; small lab tests)
Class Insecta, Order Coleoptera, Family Chrysomelidae

Delia radicum (Linnaeus)
Common name: cabbage root maggot (adult only, in preliminary tests on insects from canola fields)
Class Insecta, Order Diptera, Family Anthomyiidae

Sitona lineatus (Linnaeus)
Common name: pea weevil (adult; small lab tests in collaboration with Dr. H. Carcamo)
Class Insecta, Order Coleoptera, Family Curculionidae

Melanoplus bivittatus Say (lab and field; extensive testing)
Common name: two-striped grasshopper (adult and nymph)
Class Insecta, Order Orthoptera, Family Acrididae

Melanoplus packardii Scudder
Common name: Packard's grasshopper (adult and nymph)
Class Insecta, Order Orthoptera, Family Acrididae

 

Entz, S.C., Kawchuk, L.M. and Johnson, D.L. 2008. Discovery of a North American genetic variant of the entomopathogenic fungus Metarhizium anisopliae var. anisopliae pathogenic to grasshoppers. BioControl 53(2): 327-339. (Full paper)

Entz, S.C., Johnson, D.L., and Kawchuk, L.M. 2005. Development of a PCR-based diagnostic assay for the specific detection of the fungus Metarhizium anisopliae var. acridum in grasshoppers. Mycological Research 109: 1302-1312.

Johnson, D.L., Smits, J.S., Jaronski, S.T., and Weaver, D.K. 2002. Assessment of health and growth of ring-necked pheasants following consumption of infected insects or conidia of entomopathogenic fungi, Metarhizium anisopliae var acridum and Beauveria bassiana, from Madagascar and North America. Journal of Toxicology and Environmental Health 65: 2145-2162.

Lomer, C.J., Bateman, R.P., Johnson, D.L., Langewald, J., and Thomas, M.B. 2001. Biological control of locusts and grasshoppers. Annual Review of Entomology 46: 667-702.

Smits, J.E., Johnson, D.L., and Lomer, C. 1999. Pathological and physiological responses of ring-necked pheasant chicks following dietary exposure to the fungus Metarhizium flavoviride, a biocontrol agent for grasshoppers in Africa. Journal of Wildlife Diseases 35: 194-203.