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APPENDIX 2 - Research Protocol

This appendix is for reference purposes only and is not a prescriptive part of the standard.


Research materials

It is recommended to archive samples of the different developmental stages of the pests studied in order to, among other reasons, resolve possible future disputes on identification. The commodity to be used should be of normal commercial condition.

To perform treatment research to control quarantine pests it is necessary to know its basic biology as well as define how the pests used in the research will be obtained. The experiments with irradiation should be carried out on the commodity infested naturally in the field and/or with laboratory-reared pests that are used to infest the commodity preferably in a natural form. The method of rearing and feeding should be carefully detailed.

Note: Studies done with pests in vitro are not recommended because the results could be different from those obtained when irradiating the pests in commodities unless preliminary testing indicates that results from in vitro treatments are no different than in situ.


The dosimetry system should be calibrated, certified and used according to recognized international standards. The minimum and maximum doses absorbed by the irradiated product should be determined striving for dose uniformity. Routine dosimetry should be conducted periodically.

International ISO Guidelines are available for conducting dosimetry research on food and agricultural products (see Standard ISO/ASTM 51261 Guide for Selection and Calibration of Dosimetry Systems for Radiation Processing).

Estimation and confirmation of minimum absorbed dose for treatment

Preliminary Tests

The following steps should be carried out to estimate the dose required to ensure quarantine security:

· Radiosensitivity of the different stages of development of the pest in question that may be present in the commodity that is marketed must be established with the purpose of determining the most resistant stage. The most resistant stage, even if it is not the most common one occurring in the commodity, is the stage for which the quarantine treatment dose is established.

· The minimum absorbed dose will be determined experimentally. If pertinent data do not already exist, it is recommended to use at least five (5) dose levels and a control for each developmental stage, with a minimum of 50 individuals where possible for each of the doses and a minimum of three (3) replicates. The relationship between dose and response for each stage will be determined to identify the most resistant stage. The optimum dose to interrupt the development of the most resistant stage and/or to avoid the reproduction of the pests needs to be determined. The remainder of the research will be conducted on the most radiotolerant stage.

· During the period of post-treatment observation of the commodities and associated pests, both treated and control, must remain under favorable conditions for survival, development, and reproduction of the pests so that these parameters can be measured. The untreated controls must develop and/or reproduce normally for a given replicate for the experiment to be valid. Any study where the control or check mortalities are high indicates that the organisms were held and handled under sub-optimal conditions. These organisms may give misleading results if their treatment mortality is used to predict an optimum treatment dose. In general, mortality in the control or check should not exceed 10%.

Large Scale (Confirmatory) Tests

· To confirm if the estimated minimum dose to provide quarantine security is valid, it is necessary to treat a large number of individuals of the most resistant stage of the organism while achieving the desired result, be it prevention of pest development or sterility. The number treated will depend on the required level of confidence. The level of efficacy of the treatment should be established between the exporting and importing countries and be technically justifiable.

· Because the maximum dose measured during the confirmatory part of the research will be the minimum dose required for the approved treatment, it is recommended to keep the maximum-minimum dose ratio as low as possible.


Test records and data need to be kept to validate the data requirements and should upon request be presented to interested parties, for example the NPPO of the importing country, for consideration in establishing an agreed commodity treatment.

For further information on international standards, guidelines and recommendations concerning phytosanitary measures, and the complete list of current publications, please contact the:


By mail:

IPPC Secretariat
Plant Protection Service
Food and Agriculture Organization of the United Nations (FAO)
Viale delle Terme di Caracalla
00100 Rome, Italy






International Plant Protection Convention, 1997. FAO, Rome.

ISPM No. 1: Principles of plant quarantine as related to international trade, 1995. FAO, Rome.

ISPM No. 2: Guidelines for pest risk analysis, 1996. FAO, Rome.

ISPM No. 3: Code of conduct for the import and release of exotic biological control agents, 1996. FAO, Rome.

ISPM No. 4: Requirements for the establishment of pest free areas, 1996. FAO, Rome.

ISPM No. 5: Glossary of phytosanitary terms, 2003. FAO, Rome.

Glossary Supplement No. 1: Guidelines on the interpretation and application of the concept of official control for regulated pests, 2001. FAO, Rome.

Glossary Supplement No. 2: Guidelines on the understanding of potential economic importance and related terms including reference to environmental considerations, 2003. FAO, Rome

ISPM No. 6: Guidelines for surveillance, 1997. FAO, Rome.

ISPM No. 7: Export certification system, 1997. FAO, Rome.

ISPM No. 8: Determination of pest status in an area, 1998. FAO, Rome.

ISPM No. 9: Guidelines for pest eradication programmes, 1998. FAO, Rome.

ISPM No. 10: Requirements for the establishment of pest free places of production and pest free production sites, 1999. FAO, Rome.

ISPM No. 11 Rev. 1: Pest risk analysis for quarantine pests including analysis of environmental risks, 2003. FAO, Rome.

ISPM No. 12: Guidelines for phytosanitary certificates, 2001. FAO, Rome.

ISPM No. 13: Guidelines for the notification of non-compliance and emergency action, 2001. FAO, Rome.

ISPM No. 14: The use of integrated measures in a systems approach for pest risk management, 2002. FAO, Rome.

ISPM No. 15: Guidelines for regulating wood packaging material in international trade, 2002. FAO, Rome.

ISPM No. 16: Regulated non-quarantine pests: concept and application, 2002. FAO, Rome.

ISPM No. 17: Pest reporting, 2002. FAO, Rome.

ISPM No. 18: Guidelines for the use of irradiation as a phytosanitary measure, 2003. FAO, Rome.

ISPM No. 19: Guidelines on lists of regulated pests, 2003. FAO, Rome.

[5] Based primarily on insect pest treatment research.

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