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29 BIOASSAYS FOR SCREENINGS AND QUALITY CONTROL OF PRODUCTS WITH INSECTICIDAL, FUNGICIDAL AND NEMATOCIDAL ACTIVITY, BASED ON PLANT EXTRACTS, MICROORGANISMS AND CHEMICAL MOLECULES M. Ricci, G. Flek, M. Colli, R. Barcarotti, L. Quattrocchi, S. Coranelli, L. Concezzi, A.P. Fifi, R. De Nicola, M. Scribano, M. De Berardinis, F. Citarrei and A. Ragni Introduction BioTecnologie BT srl Pantalla di Todi 06050 Perugia Italy Tel: ++39 – 075 - 895091 Fax: ++39 – 075 – 888 776 E-mail: mricci.bt@parco3a.org A bioassay is an experiment where a living organism (insect, plant, phytopathogenic nematode and others) is used as test (Robertson J.L. & Preisler H.K.). It is a fundamental tool in the R & D of agricultural products. Bioassays are used to discover new molecules and/or toxins, to compare different pesticides, to evaluate the feasibility of excipients for formulations, attractants and/or repellents, and for the quality control of products ready for the market. Does not exist a universal bioassay: for each purposes a specific bioassy should be used or, if not available, should be developed (Ricci et al., 1996). A good bioassay should be cheap, fast, should allow the repeatability of the results in different dates and different sites, should discriminate two products with little differences, and should predict, as far as possible, the further performance of the product in field conditions. Here, are presented a series of bioassays that are used, adapted and developed by the company BioTecnologie B.T. S.r.l., for its R&D programs or for services for third parties. Bioassays Commercial companies that produce and sell bio-products for agricultural use, based on Entomopathogenic Nematodes, Fungi, Bacteria and Viruses, have to set up a bioassay unit in order to perform screening programs and to carry out quality control of the production and quality control of final products. The bioassay should be a compromise between the need to assure scientific results and the need to save time and money. To support a rational bioassay unit, other sections are needed: Insectary, Plant Pathogen Maintenance and Greenhouses. Screenings Screenings are performed in order to discover new active ingredients or to support the development of products. According to the different needs, different bioassays are designed. At BioTecnologie B.T., since 1992, several procedures for bioassays have been developed. Toxicity bioassays and feeding behavior tests are performed daily for inside and outside (external service) research purposes. Quality Control of Bio-products for agricultural pest control The first duty of the quality control system is to assess what is declared on the label of the product in terms of product weight and titer of active ingredient. Even if bio-products are based on non-pathogenic (to human beings) microorganisms, the producer has to assure that his product is free of pathogenic contaminants (Ricci and Fridlender, 1998). Stability is an important parameter in terms of maintenance of the product characteristics during storage and transportation. The activity is the parameter that shows the ability of the product to control the target insects in real conditions. This is the harder parameter to assess because a simple, cheap, fast and reliable laboratory bioassay has to be found in order to predict the activity of the product under field conditions Insectary A multiple-species insect rearing facility for research support on bio-insecticide development is operating at BioTecnologie B.T. S.r.l., in Umbria – Italy, since 1992. PDF creato con FinePrint pdfFactory versione dimostrativa http://www.secom.re.it/fineprint 136
Several colonies of Lepidopteran, Coleopteran and Dipteran species have been established and are maintained on a regular basis whereas other species are reared on a temporary basis when needed. Standardized rearing procedures have been developed that ensure consistent quality of the test insects. The following species are currently reared: Lepidopteran: Agrotis ipsilon��Mamestra brassicae, Spodoptera littoralis, S. exigua, Helicoverpa armigera, Lobesia botrana, Cydia molestaColeopteran: Otiorhynchus sp., Tenebrio molitorDipteran: Culex pipiens pipiens Plant Pathogen Colony Maintainance Several pathogens are maintained regularly in vitro to support the research and development of bio-phytosanitary products. The following fungi species are currently maintained: Fusarium culmorum, F. crookwellense, F. oxysprum dianthii, F. sambucinum, F. solani, F. graminearum, F. akuminatum, F. oxysporum, Rhizoctonia solani, Alternaria solani, Botrytis cynerea, Penicillum italicum, Cercospora beticola, Phytophthora cinnamoni, Pyranochaeta lycopersici, Phoma spp., Microdochium nivale The following phytopathogenic nematode species is currently reared: Meloidogyne incognita Greenhouses Modern and well-equipped greenhouses support the growth of different plant species for insect and phytopathogenic nematode rearing, pathogen maintenance and for glasshouse assays. The following plants have been grown: Corn, Melon, Tomato, Pepper, Cyclamen, Impatient, Tobacco, Celery, Rapeseed . Conclusions: Due to its facility and its experience, BioTecnologie B.T., is able to design, develop and perform several bioassays for the development and the quality control of pesticides and biopesticides. References Ricci M., Glazer I. and Gaugler R., 1996. Comparison of Bioassays to Measure Virulence of Different Entomopathogenic Nematodes. Biocontrol Science and Technology (1996) 6, 235-245 Ricci M. and Fridlender B., 1998. Quality Control of a Biological Insecticide Based on Entomopathogenic Nematodes: NEMA-BIT. At Gent, B: Cost 819 Workshop on Quality Control of Entomopathogenic Nematodes. p. 16 and 34-35 Robertson J.L. &. Priesler H.K, 1992. Pesticide bioassays with arthropods. CRC Press, Boca Raton. Pictures Bioassay Tray: most of the screenings for micro-organisms and molecules against target insects are carried out in this particular tray where individual insect is tested on an artificial diet PDF creato con FinePrint pdfFactory versione dimostrativa http://www.secom.re.it/fineprint 137
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International Congress BIOLOGICAL P
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15:40-16:20 G. Lozzia Università d
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MAIN SPEAKERS • V. Rotondo FIAO,
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Il settore oggi è regolamentato in
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target botanical synthetic sodium c
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RELIABILITY AND PROSPECTIVES FOR TH
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Vineyard Passive diffusion Spontane
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of the host, a relatively limited n
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LOZZIA G.C., RIGAMONTI I.E., 1994 -
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DIFESA DELLE COLTURE DAI PATOGENI F
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THE FUNGAL PHYTOTOXINS AND THEIR PO
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NEED FOR RESEARCH AND APPROPRIATE A
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PRESENCE OF OCHRATOXIN IN EXPERIMEN
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5. Hohler, D. (1998). Ochratoxin A
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Table 3: Pesticide treatments on
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BIOPESTICIDES: EVALUATION PROCESS F
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Xenorhabdus spp. and Photorhabdus s
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It can be reproduced in vivo at 15
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Vinson, S. B., 1975, Biochemical co
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PROTECTION OF GRAPEVINE FROM POWDER
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I MEZZI TECNICI PER L'AGRICOLTURA B
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POSTER 1. Agosteo G.E., Ambrosio M.
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31. Pietri A., Bertuzzi T., Barbier
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1 PROVE DI LOTTA ALL’OIDIO DELLO
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2 LUPINUS ALBUS, AN ANCIENT EUROPEA
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Table 1. Benzo(a)pyrene and benzo(e
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5 INSETTICIDI D’ORIGINE VEGETALE
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Tabella 2. Piretrine: composizione
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californicus (fitoseide predatore)
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7 NATURAL COMPOUNDS IN THE CONTROL
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8 METHODS FOR EVALUATION, IN CONTRO
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Figure 3. Effect of Pseudomonas sp.
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9 IMPROVEMENT OF SOIL PROPERTIES AN
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(mg kg-1) (mg kg-1) 140 120 100 80
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10 NEMATICIDAL ACTIVITY OF AQUEOUS
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% cumulative hatch 40 35 30 25 20 1
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ROTENONE Il principio attivo è est
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- azadiractina nei confronti di F.
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Recovery and repeatabilty tests wer
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13 IMPIEGO DEL TIMOLO NEL CONTROLLO
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dello standard esterno. E’ stata
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Tab. III - Residui di timolo(mg/Kg)
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Page 87 and 88: 15 ORGANIC WHEAT QUALITY AND PRODUC
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Page 95 and 96: Obtained recoveries percent were: 8
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Page 129 and 130: Results and Discussion S. carpocaps
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