Publications


  • 8-September-2009

    English

    Test No. 403: Acute Inhalation Toxicity

    This method provides information on health hazard likely to arise from short-term exposure to a test article (gas, vapour or aerosol/particulate test article) by inhalation.

    The revised Test Guideline describes two studies: a traditional LC50 protocol and a Concentration x Time (C x t) protocol. It can be used to estimate a median lethal concentration (LC50), non-lethal threshold concentration (LC01) and slope, and to identify possible sex susceptibility. This Test Guideline enables a test article quantitative risk assessment and classification according to the Globally Harmonized System for the Classification and Labelling of Chemicals. In the traditional LC50 protocol, animals are exposed to one limit concentration or to three concentrations, at least, for a predetermined duration, generally of 4 hours. Usually 10 animals should be used for each concentration. In the C x T protocol, animals are exposed to one limit concentration or a series of concentrations over multiple time durations. Usually 2 animals per C x t interval are used. Animals (the preferred species is the rat) should be observed for at least 14 days. The study includes measurements (including weighing), daily and detailed observations, as well as gross necropsy.

  • 8-September-2009

    English

    Test No. 438: Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants

    The Isolated Chicken Eye  (ICE) test method is an in vitro test method that can be used to classify substances as “ocular corrosives and severe irritants”. The ICE method uses eyes collected from chickens obtained from slaughterhouses where they are killed for human consumption, thus eliminating the need for laboratory animals. The eye is enucleated and mounted in an eye holder with the cornea positioned horizontally. The test substance and negative/positive controls are applied to the cornea. Toxic effects to the cornea are measured by a qualitative assessment of opacity, a qualitative assessment of damage to epithelium based on fluorescein retention, a quantitative measurement of increased thickness (swelling), and a qualitative evaluation of macroscopic morphological damage to the surface. The endpoints are evaluated separately to generate an ICE class for each endpoint, which are then combined to generate an Irritancy Classification for each test substance.

  • 8-September-2009

    English

    Test No. 509: Crop Field Trial

    Crop field trials are conducted to determine the magnitude of the pesticide residue in or on raw agricultural commodities, including feed items, and should be designed to reflect pesticide use patterns that lead to the highest possible residues.

    Objectives of crop field trials are to: (1) quantify the expected range of residue(s) in crop commodities following treatment according to the proposed or established good agricultural practice; (2) determine, when appropriate, the rate of decline of the residue(s) of plant protection product(s) on commodities of interest; (3) determine residue values such as the “Supervised Trial Median Residue” and “Highest Residue” for conducting dietary risk assessment; and (4) derive maximum residue limits (MRLs).  This Test Guideline requires one sample from treated plots at each sampling interval for crops that have eight or more crop field trials.

    The test substance(s) should be stored under appropriate conditions for the study duration and applied soon after preparation or mixing. Test substance applications should not be made in strong wind, during rain or when rainfall is expected shortly after application. For all applications, the application rate should be expressed in terms of amount of product and/or active ingredient per unit area. At the end of each crop field trial, the (stored) samples are analysed for residue level (expressed for example in mg/kg).

  • 8-September-2009

    English

    Test No. 452: Chronic Toxicity Studies

    The objective of these chronic toxicity studies is to characterize the profile of a substance in a mammalian species (primarily rodents) following prolonged and repeated exposure.
    The Test Guideline focuses on rodents and oral administration. Both sexes should be used. For rodents, at least 20 animals per sex per group should normally be used at each dose level, while for non-rodents a minimum of 4 per sex per group is recommended. At least three dose levels should be used in addition to the concurrent control group. Frequency of exposure normally is daily, but may vary according to the route chosen (oral, dermal or inhalation) and should be adjusted according to the toxicokinetic profile of the test substance. The duration of the exposure period should be 12 months. The study report should include: measurements (weighing) and regular detailed observations (haematological examination, urinalysis, clinical chemistry), as well as necropsy procedures and histopathology.

  • 8-September-2009

    English

    Test No. 437: Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants

    The Bovine Corneal Opacity and Permeability test method (BCOP) is an in vitro test method that can be used to classify substances as 'ocular corrosives and severe irritants'. The BCOP uses isolated corneas from the eyes of cattle slaughtered for commercial purposes, thus avoiding the use of laboratory animals. Each treatment group (test substance, negative/positive controls) consists of a minimum of three eyes where the cornea has been excised and mounted to a holder. Depending on the physical nature and chemical characteristics of the test substance, different methods can be used for its application since the critical factor is ensuring that the test substance adequately covers the epithelial surface. Toxic effects to the cornea are measured as opacity and permeability, which when combined gives an In Vitro Irritancy Score (IVIS) for each treatment group. A substance that induces an IVIS superior or equal to 55.1 is defined as a corrosive or severe irritant.

  • 8-September-2009

    English

    Test No. 229: Fish Short Term Reproduction Assay

    This Test Guideline describes an in vivo screening assay for fish reproduction where sexually mature male and spawning female fish are held together and exposed to a chemical during a limited part of their life-cycle (21 days). The short term reproduction assay was validated in the fathead minnow (Pimephales promelas) and this is the recommended species. The assay is run with three test chemical concentrations and the necessary controls, including a carrier control if necessary. For the fathead minnow, four replicate test vessels are used for each treatment level and control(s). During the conduct of the assay, the egg production is measured quantitatively daily in each test vessel. At termination of the 21-day exposure period, two biomarker endpoints are measured in males and females separately, as indicators of endocrine activity of the test chemical; these endpoints are vitellogenin and secondary sexual characteristics. Gonads of both sexes are also preserved and histopathology may be evaluated to assess the reproductive fitness of the test animals and to add to the weight of evidence of other endpoints.

  • 8-September-2009

    English

    Test No. 453: Combined Chronic Toxicity/Carcinogenicity Studies

    The objective of a combined chronic toxicity/carcinogenicity study is to identify carcinogenic and the majority of chronic effects, and to determine dose-response relationships following prolonged and repeated exposure.

    The rat is typically used for this study. For rodents, each dose group and concurrent control group intended for the carcinogenicity phase of the study should contain at least 50 animals of each sex, while for the chronic toxicity phase of the study should contain at least 10 animals of each sex.  At least three dose levels should be used, in addition to the concurrent control group for both the chronic toxicity phase and the carcinogenicity phase of the study. The three main routes of administration are oral, dermal, and inhalation. The Test Guideline focuses on the oral route of administration.

    The period of dosing and duration of the study is normally 12 months for the chronic phase, and 24 months for the carcinogenicity phase. The study report should include:  measurements (weighing) and regular detailed observations (haematological examination, urinalysis, clinical chemistry), as well as necropsy procedures and histopathology. All these observations permit the detection of neoplastic effects and a determination of carcinogenic potential as well as the general toxicity.

  • 8-September-2009

    English

    Test No. 302C: Inherent Biodegradability: Modified MITI Test (II)

    This Test Guideline describes the modified MITI test (II). This test permits the measurement of the Biochemical Oxygen Demand (BOD) and the analysis of residual chemicals in order to evaluate the inherent biodegradability of chemical substances which have been found by the Standard MITI Method (I) to be low degradable.

    An automated closed-system oxygen consumption measuring apparatus (BOD-meter) is used. Chemicals to be tested are inoculated in the testing vessels (six bottles with different quantities of test chemical) with micro-organisms. In order to check the activity of the inoculum, the use of control substances (aniline, sodium acetate or sodium benzoate) is desirable. During the test period, the BOD is measured continuously. Biodegradability is calculated on the basis of BOD and supplemental chemical analysis, such as measurement of the dissolved organic carbon concentration, concentration of residual chemicals, etc. The BOD curve is obtained continuously and automatically for 14 to 28 days. After the 14 to 28 days of testing, pH, residual chemicals and intermediates in the testing vessels are analysed.

  • 8-September-2009

    English

    Test No. 231: Amphibian Metamorphosis Assay

    This Test Guideline describes an amphibian metamorphosis assay intended to screen substances which may interfere with the normal functioning of the hypothalamo-pituitary-thyroid axis. The assay was validated with the species Xenopus laevis, which is recommended for use in the Guideline. The assay uses three test chemical concentrations and the necessary controls, including a carrier control if necessary. The assay starts with tadpoles at the development stage 51 on the Nieuwkoop and Faber scale and is extended for a duration of 21 days. Four replicate test vessels are used for each treatment level and control(s). After 7 days of exposure, a sub-set of tadpoles from each treatment level is sampled for the measurement of the length of the hind-limb. At termination of 21-day exposure period, developmental stage, snout-to-vent length and hind limb length are measured on all remaining tadpoles. A sub-set of tadpoles from each treatment level is fixed (whole-body or dissected) for histopathology of the thyroid gland.

  • 8-September-2009

    English

    Energy Policies of IEA Countries: Portugal 2009

    The International Energy Agency's 2009 review of Portugal's energy policies and programmes.  This edition finds that Portugal has made considerable efforts to strengthen its energy policy since the last IEA in-depth review in 2004. A large number of IEA recommendations have been successfully implemented, including greater diversification of the energy mix and increased energy policy co-ordination. A new National Energy Strategy, published in October 2005, identified three principal means for meeting Portugal’s policy goals: the promotion of renewable energy, increased energy efficiency and competition in energy markets.

    Over a short period of time, Portugal has become a leader in terms of renewable energy development.  Well-designed incentive mechanisms and the adoption of ambitious targets ensure hydro, wind and other technologies will continue to grow. The National Action Plan for Energy Efficiency was enacted in 2008, and Portugal aims to implement energy efficiency measures equivalent to 9.8% of total final energy consumption by 2015. This plan complements a well developed and co-ordinated climate change policy. Further steps have been taken towards the liberalisation of energy markets, including the innovative creation of a single operator for the transport of natural gas and electricity, natural gas storage and operation of the Sines LNG terminal.

    Still, a number of challenges remain. Energy markets are not as competitive as policy makers may have wished, and energy research and development policy coordination needs to be strengthened.

    This review provides sectoral critiques of existing policy and recommendations for further improvements. It is intended to serve as an indispensable guide for Portuguese policy makers as they travel along the path to a more sustainable energy future.

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