Guidelines for Risk Assessment
Annex 3b to the Danish Working Environment Authority's Executive Order No. 910 of 11 September 2008 on Genetic Engineering and Working Environment.
Annex 3a, Part A, Points 1 and 2 sets out the elements to be taken into account when assessing potentially adverse effects on human health and the environment. Potentially adverse effects are defined as: effects that cause disease or render prophylaxis or treatment ineffective; adverse effects on organisms or natural populations due to establishment and/or spreading in the environment; or adverse effects due to gene transfer to other organisms. The risk of these potentially adverse effects must be examined for each operation to be classified into a given class, taking into account the nature and scope of the operation, in order to determine the necessary containment measures. The risk of contained use of genetically modified organisms (GMOs) and of the construction of such organisms is determined by examining the severity of the potential harmful effect on human health or the environment and the likelihood of these effects occurring. In the risk assessment, human or environmental exposure to GMOs is assessed as a result of the operation of an enclosed-use facility or as a result of potential unintended release from such a facility. The classification level determined in the risk assessment determines the containment requirements that apply to activities involving GMOs.
2. Risk Assessment
A complete risk assessment process involves the following two procedures:
2.1 Procedure 1
Identification of potential harmful properties (hazards) of the GMO and placement of the GMO into a provisional class (Classes 1 to 4) based on the severity of the potential adverse effects;
assessment of the likelihood of adverse effects occurring, based on an assessment of the potential exposure (both human and environmental) taking into account the nature and scope of the activity in question and the containment measures for the provisional class in which the activity is placed.
2.2 Procedure 2
Final classification and final determination of containment measures.
The appropriateness of the final classification and the final containment measures are confirmed by repeating Procedure 1.
3. Procedure 1
3.1 Identification of harmful properties (hazards) of the GMO.
The risk assessment process includes an identification of any harmful properties of the GMO as a result of the genetic modification or any changes in the properties of the recipient organisms. The potentially harmful properties associated with the GMO must be determined by examining the recipient organism, the donor organism, the properties and location of the inserted genetic material and any vector. It is important to keep in mind that genetic modification of an organism can affect its ability to harm human health and the environment, as it may decrease, increase or remain unchanged.
3.2 Aspects that should be considered, if applicable
3.2.1 Recipient organism
- Name, designation and origin;
- Information about the parental organism(s) or, where applicable, the reproductive cycle of the host organism (gender/gender-neutral);
- The characteristics of pathogenicity and virulence, infectivity, allergenicity, toxicity and vectors of disease transmission;
- The characteristics of indigenous vectors and accompanying agents if these could mobilise the inserted gene material, and the frequency of such mobilisation;
- The nature and stability of any inactivating mutations;
- The stability of the parental organism(s) or host organism in terms of relevant genetic characteristics;
- Any prior genetic modifications;
- Host spectrum (if applicable);
- Important physiological characteristics that could be altered in the final GMO and their stability, if applicable;
- Natural habitat and geographical distribution;
- Significant involvement in environmental processes (such as nitrogen fixation or pH regulation);
- Interaction with and impact on other organisms in the environment (including likely competitive, pathogenic or symbiotic properties);
- Ability to form survival structures (such as spores or sclerotia).
3.2.2 Donor organism
- Name, designation and origin;
- Degree of relatedness to the host organism;
- Characteristics of pathogenicity and virulence, infectivity, toxicity and vectors of disease transmission;
- Characteristics of indigenous vectors:
- Frequency of mobilisation as well as specificity;
- Presence of genes conferring resistance to antimicrobial agents, e.g. antibiotics;
- Host spectrum;
- Other relevant physiological characteristics.
- Description of the modification, including the method of vector insertion into the host organism or the method used to produce the genetic modification in question;
- The specific identity and function of the insert (genes);
- The expression level and rate of the inserted genetic material; Measurement method and sensitivity;
- The origin of the genetic material, the identity and characteristics of the donor organism(s), where applicable;
- Previous genetic modifications, if applicable;
- The location of the inserted genetic material (the possibility of activation/deactivation of host genes as a result of the insertion).
- Characteristics of the vector, its origin;
- Structure and amount of nucleic acids from vector and/or donor that may remain in the final construction of the modified micro-organism;
- If the inserted vector is present in the final GMO: frequency of mobilisation and/or ability to transfer genetic material.
3.2.5 The final GMO
- The stability of the organism in terms of genetic characteristics;
- Characteristics of substances that are formed or may be formed as a result of the genetic manipulation;
- The activities of the expressed protein;
- Factors that may affect the survival, reproduction and spreading of the modified organism outside the nutrient broth.
220.127.116.11 Health aspects
- Expected toxic or allergenic effects of the GMO and/or its metabolic products;
- Product risks, including the formation of toxins or biologically highly active substances;
- Comparison between the modified micro-organism and the recipient organism or (if applicable) the parental organism in terms of pathogenicity;
- Expected colonisation ability;
- Known and expected habitats.
- If the micro-organism is pathogenic to immunocompetent humans:
- Diseases caused and the mode of transmission, including invasiveness and virulence;
- Infectious dose;
- Possible change of infection pathway or tissue specificity;
- Possibility of survival outside human host;
- Presence of vectors or other modes of propagation;
- Biological stability;
- Antibiotic resistance patterns;
- Possibility of appropriate disease treatment and prophylaxis.
18.104.22.168 Environmental considerations
- Ecosystems into which the organism could accidentally be released from the contained use;
- Expected survivability, reproduction and spreading of the modified organism in the identified ecosystems;
- Expected result of interaction between the modified organism and the organisms or micro-organisms that may be exposed in the event of unintended release into the environment;
- Known or anticipated effects on plants and animals, such as pathogenicity, toxicity, allergenicity, pathogen carrier, altered antibiotic resistance patterns, altered tropism or host specificity, colonisation;
- Known or anticipated participation in biogeochemical processes.
22.214.171.124 Monitoring techniques
- Techniques for the detection, identification and monitoring of the modified organism;
- Techniques for detecting the transfer of the new genetic material to other organisms;
- Possible methods of decontamination of the area in case of unintended release.
3.3 Provisional classification of the GMO
Annex 3a, Part A, Points 3-5, states that the first stage of the risk assessment of a GMO consists in identifying the potential harmful properties of the GMO so that a provisional classification can be made. This is done by identifying the hazards that may be associated with recipient organism, donor organism, vector and insert. The assessment may take into account the general characteristics of Class 1, as described in Annex 3a, Part A, Point 4, and appropriately updated national and international classification schemes (including the Executive Order Concerning the Protection of Workers from Risks Related to Exposure to Biological Agents at Work as amended). The corresponding set of containment and other protective measures listed in Annex 2 serves as a basis for assessing whether more stringent containment and control measures are required to control the proven adverse effects.
The risk of adverse effects due to any harmful properties of the GMO is assessed by examining the severity of the harm and any biological properties (e.g. inactivating mutations) which limit the likelihood of harm occurring. The severity of the adverse effects is assessed independently of the likelihood of their occurrence. The seriousness of any harm is assessed by examining what the consequences could be and not the probability that the harm will occur in the case in question. For a pathogen, for example, it should be assessed how severe the disease would be if a susceptible species became infected. When placing a GMO into a provisional class, the severity of the harm is also taken into account. In the various classification schemes, such as in the Executive Order Concerning the Protection of Workers from Risks Related to Exposure to Biological Agents at Work, the seriousness of any adverse effects has been taken into account. However, many of these schemes are based solely on either human health or environmental considerations. It is important to ensure that the seriousness of the harmful human health and environmental effects of the GMO in question are taken fully into account.
3.4 Assessment of the likelihood of adverse effects occurring.
The likelihood of a harmful event occurring depends first and foremost on the level and nature of human or environmental exposure to a given GMO. In most cases, the exposure aspect is of primary importance for the risk assessment, as it is often decisive as to whether a adverse effect will occur. The likelihood of human or environmental exposure to a GMO depends on the operations carried out (e.g. and on what scale) and the containment measures taken, based on the provisional classification according to Points 5 and 6.
In accordance with Point 7 (Nos. ii) and iii)) of Annex 3a, the final classification and choice of control measures shall take into account the characteristics of the operation. The nature and scope of the activity must be taken into account in order to assess the likelihood of human and environmental exposure, and this will also have an impact on the choice of appropriate risk management procedures.
Among the characteristics of the operation that may be relevant to the risk assessment and which should therefore be taken into account are the activity itself, work routines, the scope of the activity and the containment measures applied.
In the assessment, special consideration must be given to how waste and run-off water are to be disposed of. Where appropriate, safety measures must be taken to protect human health and the environment.
3.4.1. Nature of proposed activities
The level of risk and the application of control measures to reduce this risk from the GMO to an appropriate level depend on the nature of the activities, as these will have an impact on human and environmental exposure and thus on the likelihood of adverse effects occurring.
The nature of the work will also determine which table in Annex 2 contains the most appropriate containment and control measures.
For laboratory-scale work where the effects of the standard laboratory procedures on exposure are well known, a detailed risk assessment of each procedure will likely not be needed unless an extremely dangerous organism is used. However, a more detailed assessment may be necessary for non-routine procedures or procedures that may have a significant impact on the level of risk, e.g. procedures by which aerosols are generated.
3.4.2 Concentration and scope
High cell density can lead to the risk of exposure to high concentrations of the GMO, especially in downstream processing operations. The significance of the concentration for the probability of a harmful event occurring must be investigated.
The scope of an activity must also be taken into account in the risk assessment, both the scope of a single operation or in the form of frequent repetition of a process, because both can increase the likelihood of exposure if containment and control measures fail and thus increase the likelihood of a harmful event.
Although large-scale operations do not necessarily mean high risk, increased scope may increase the likelihood of exposure both in terms of the number of people and the scope of the environment exposed in the event of failure of containment measures.
The scope of the work will also have an impact on which table in Annex 2 contains the most appropriate containment and control measures.
3.4.3 Cultivation conditions
For many activities with contained use, the cultivation is subject to strict containment conditions, but the design and nature of the reactors and other cultivation equipment also have an impact on the risk to human health and the environment. The exposure to, and thus the risk of a GMO, can be significantly reduced with the help of high-tech and tightly closed fermentation tanks. It is important to investigate the reliability of this equipment and the possibility of malfunctions in cases where malfunctions may result in extensive exposure to harmful GMOs. Where such unintended release can reasonably be foreseen, additional containment measures may be required. The standard work routines for persons working with cultivated GMOs, such as centrifugation or sonication, are of great importance for the effectiveness of containment measures.
In addition to the physical cultivation conditions that act as containment measures, biological and chemical measures that serve to protect the operation can also significantly contribute to the containment measures. Examples of biological containment are auxotrophic mutants that require the addition of specific growth factors in order to develop. Examples of chemical containment measures are disinfectants in the discharge systems.
Point 7 of Annex 3a states that, in assessing the likelihood of adverse effects occurring and their severity, the characteristics of the environment which may be exposed should be taken into consideration as should the severity of the effects.
Certain aspects of these environmental considerations are important, such as the scope and nature of the environmental exposure and whether there is biota in the exposed area that can be damaged by the GMO in question.
If necessary, the following factors should be taken into account when assessing the impact of the characteristics of the recipient environment on the likelihood of the potential adverse effect occurring, and thus on the level of risk and on the choice of control measures.
126.96.36.199 Environment that could be exposed
In most cases, the environment that could be exposed can be expected to be limited to the workplace and the area immediately surrounding the facility but, depending on the special features of the contained use and the facility, it is also necessary to take the environment into account in a broader sense. The extent of the environmental exposure may depend on the nature and scope of the activity, but all conceivable forms of transfer in the broader environment, e.g. physical routes (such as sewers, streams, landfills, wind) and biological vectors (such as through infected animals and insects) must be taken into account.
188.8.131.52 The presence of sensitive species
The likelihood of harmful effects occurring depends on the presence of sensitive species in the environment which may be exposed, including humans, animals and plants.
184.108.40.206 The possibility that the environment may promote the survival of the GMO
An important aspect of the risk assessment is the question of the extent to which the GMO can survive and persist in the environment. The likelihood of adverse effects occurring is far lower if a GMO is unable to survive in the environment into which it could accidentally be released.
220.127.116.11 Effects on the physical environment
In addition to the direct adverse effects of a GMO, indirect adverse effects due to significant changes in the physicochemical properties of and/or the ecological balance of the environmental media, soil and water must also be taken into account.
4. Procedure 2
4.1 Determination of final classification and final containment measures.
Once the seriousness of all potentially harmful properties of the GMO and the likelihood of harm as a result have been examined, taking into account the containment and control measures provided for in the provisional classification, the definitive classification and the final containment measures for the GMO can be determined. In assessing the final classification and the final containment measures, the provisional classification should be revised to see if it was correct, taking into account the proposed activities and operations. A comparison between the provisional classification and the associated containment measures with the final class and the associated containment requirements can lead to three results:
- The provisional classification does not take due account of certain adverse effects, and the provisional containment under Procedure 1 is therefore inadequate; further containment measures are therefore required and the classification of the activity in question may need to be revised
- The provisional classification was appropriate, and the containment measures associated with it were adequate to prevent or minimise damage to human health and the environment
- The provisional classification is stricter than necessary for the activity, for which reason a lower classification with associated containment measures should be chosen
4.2 Confirmation of the appropriateness of the final containment measures.
Once the final classification and the final containment conditions have been determined, the level of human and environmental exposure should be reassessed (Procedure 1). In doing so, it must be confirmed that the likelihood of adverse effects occurring is acceptable, taking into account the nature and scope of the work and the proposed containment conditions. Once this is done, the risk assessment process is complete.
Section 6, Subsection 5 states that the assessment must be updated when there are changes in the work, working methods, work processes, etc., which are important for the company's working environment and for the external environment, if:
- If the protective measures applied are no longer adequate or the class in which the activity takes place is no longer the right class; or
- If there is reason to believe that the assessment is no longer adequate in the light of the latest scientific or technical knowledge.
Any changes in the containment conditions as a result of the revision of the risk assessment must be implemented immediately for the sake of adequate protection of human health and the environment.
The contained uses are classified into Classes 1 to 4 based on the classification as well as containment and control measures which, in the risk assessment, are found to be necessary for the adequate containment of the GMOs in connection with the proposed activities. Annex 2 describes the containment and control measures for each class.
The administrative requirements are determined based on the classification of the contained uses of GMOs.
If there is uncertainty regarding the final classification and the final containment conditions, the Danish Working Environment Authority should be contacted.