Risk Assessment of LMOs - Training Manual

Module 3: Conducting the Risk Assessment

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1. Introduction

Risk assessment is a process intended to calculate or estimate the risk to a given target organism, system, or (sub)population, including the identification of uncertainties, following exposure to a particular agent, taking into account the inherent characteristics of the agent of concern as well as the characteristics of the specific target system (WHO, 2004).

The risk assessment process involves a critical review of available data for the purpose of identifying and possibly quantifying the risks resulting from, for example, natural events (flooding, extreme weather events, etc.), technology, agricultural practices, processes, products, agents (chemical, biological, radiological, etc.) and any activity that may pose threats to ecosystems, animals and/or people.

The objective of a risk assessment under the Cartagena Protocol “is to identify and evaluate the potential adverse effects of living modified organisms on the conservation and sustainable use of biological diversity in the likely potential receiving environment, taking also into account risks to human health” (Annex III).

The results of risk assessments of living modified organisms (LMOs) are typically used by decision-makers to make informed decisions regarding the approval, with or without conditions (e.g. requirements for risk management and monitoring strategies) or prohibition of a certain use of the LMO.

	Figure 1 – Assessing risks Source: http://www.scienceinthebox.com/en_UK/safety/riskassessment_en.html.

This module provides an introduction to risk assessment and considerations that may assist risk assessors in conducting risk assessments of LMOs that are consistent with Article 15 and Annex III of the Protocol. ¹

2. Overview of the techniques used in modern biotechnology

LMOs are most commonly developed through the use of in vitro nucleic acid techniques by inserting, deleting or modifying a gene or DNA/RNA sequence into a recipient or parental organism.

The terms genetic modification, genetic engineering, recombinant DNA and DNA manipulation are terms that apply to the direct modification of an organism’s genes. The terms genetically modified organism (GMO) as well as genetically engineered or transgenic organism are often used interchangeably for LMOs. The Cartagena Protocol emphasizes the “living” nature of the organism and products thereof, namely, processed materials that are of living modified organism origin, containing detectable novel combinations of replicable genetic material obtained through the use of modern biotechnology.

	Figure 2 – In vitro nucleic acid techniques

LMOs can also be produced through cell fusion where cells from two different organisms that do not belong to the same taxonomic family are fused resulting in an organism containing the genetic information from both parental cells. The resulting LMO may contain the complete genomes of the parental organisms or parts of their genomes. Cell fusion can be applied to bacterial, fungal, plant or animal cells, using a variety of techniques to promote fusion.

2.1 Commonly used methods for genetic modification of plants

Production of LMOs through genetic modification is a multistage process that can be achieved through a variety of methodologies. Methods commonly used in the development of LM plants can be summarized as follows. ²

Once a gene of interest has been identified and isolated from a donor organism, it is manipulated in the laboratory such that it can be inserted effectively into the intended recipient organism. The manipulation may, for example, include changes to the sequence of nucleotides so as to enhance or modulate the expression of the gene once it is introduced into the intended recipient organism.

One or more genes of interest, as well as other nucleotide sequences needed for the proper functioning of the gene(s) of interest, may then be built in an orderly sequence into a “gene construct”. The gene construct typically includes a “promoter sequence” and “termination sequence” which are necessary to ensure that the gene is expressed correctly in the recipient organism. Different promoter sequences control gene expression in different ways; some allow continuous expression of the gene (these promoters are known as “constitutive”), while others switch expression of the gene on or off at different tissues, organs and/or developmental stages of the organisms or in reaction to other external influences. Some promoters may be highly specific to the point that they drive gene expression only in a few cells of the organism and during short, specific developmental stages.

A “marker gene” is often incorporated into the gene construct to help identify which individuals of a recipient organism have been modified by the introduction of the gene construct.

Finally, the gene construct may be incorporated into a larger DNA molecule to be used as vector (e.g. plasmid, virus, etc.). The purpose of the vector is to assist the transfer of the gene construct into the recipient organism.

	Example 1 – Scheme of a gene construct and vector Note: Gene constructs currently used may include multiple elements — for example, several promoter sequences and desired genes. Source: IUCN (2003).

The recipient organism is then transformed by different methods (e.g. via infection using Agrobacterium, particle bombardment, injection, etc) for integration of the gene construct into the genome of the recipient organism.

Transformed cells are then selected and regenerated into complete LMOs. The subsequent step is the further selection of the LM event(s) that contains the desired transgene(s) and expresses the desired characteristics. Through selection, many experimental LMOs are discarded and only a few events reach the stage of commercialization.

In the case of LM plants, cross-breeding to introduce the transgene(s) into other recipient varieties is also common.

	Figure 3 – Genetic modification of plants Source: Mirkov (2003).

3. Overview of the risk assessment methodology

In order to understand what is meant by risk assessment it is important to be familiar with the concepts of risk and hazard, and how these terms differ. The term “risk” does not have a single unambiguous definition but it is often defined as “the probability of harm”. This is broadly understood as the likelihood that a harmful consequence will occur as the result of an action or condition.

Risk is often assessed through the combined evaluation of hazard and exposure.

• “Hazard” is defined as the potential of an agent or stressor to cause harm to a biological system (e.g. a species) (UNEP/IPCS, 1994).
• “Exposure” means the contact between a hazard and a receptor. Contact takes place at an exposure surface over an exposure period (WHO, 2004).

The exposure pathway from the hazard to the receptor and the possible exposure scenarios form important additional elements in understanding risk. Ascribing the probability and consequences of exposure of a receptor to the hazard characterizes the risk. All these elements must be evaluated to form an effective and useful risk assessment for specific scenarios (UNEP Division of Technology, Industry and Economics, website).

A simple example can be used to distinguish hazard from risk: acids may be corrosive or irritant to human beings (=hazard). The same acid is a risk to human health only if humans are exposed to it without protection. Thus, the degree of harm caused by the exposure will depend on the specific exposure scenario. If a human only comes into contact with the acid after it has been heavily diluted, the risk of harm will be minimal but the hazardous property of the chemical will remain unchanged (EEA, 1998).

	Example 2 – What is risk? What is risk assessment? Risk = the combination of the magnitude of the consequences of a hazard, if it occurs, and the likelihood that the consequences will occur. Risk assessment = the measures to estimate what harm might be caused, how likely it would be to occur and the scale of the estimated damage. Source: UNEP (1995).

According to WHO (2004), a risk assessment process can be divided into four main phases:

1. Hazard identification – The identification of the type and nature of adverse effects that an agent has the inherent capacity to cause on an organism, system, or (sub)population.
2. Hazard characterization – The qualitative and, wherever possible, quantitative description of the inherent property of an agent or situation that has the potential to cause adverse effects. This should, where possible, include a dose–response assessment and its attendant uncertainties.
3. Exposure assessment – Evaluation of the exposure of an organism, system, or (sub)population to an agent (and its derivatives).
4. Risk characterization – The qualitative and, wherever possible, quantitative determination, including attendant uncertainties, of the probability of occurrence of known and potential adverse effects of an agent in a given organism, system, or (sub)population, under defined exposure conditions.

If risks are identified during the last step above, risk management strategies may be identified which may effectively prevent, control or mitigate the harm from happening. As such, the risk assessment process often includes an additional step to identify a range of possible risk management strategies that could reduce the level of risk. It is worth noting, however, that it is only during the decision-making process that a choice is made as to whether or not risk management strategies should be implemented (see more details on the identification of risk management strategies in Module 4).

	Example 3 – Variation in terminology used to describe methodological components common to many risk assessment frameworks Source: Hill (2005).

As a whole the risk assessment process can be highly iterative; meaning that one or several steps may need to be re-evaluated when, for instance, new information becomes available in an attempt to increase the level of certainty.

The methodologies for risk assessment of LMOs have evolved over the last several years. At a conceptual level, the methodologies have been adapted from the existing paradigms for environmental risk assessment developed for chemicals and other types of environmental stressors (Hill, 2005). As a result, the terminology used within each methodology may vary.

Familiarity with the different terms used in risk assessment enables a more direct comparison between the terminology used in Annex III and different risk assessment frameworks. It will also facilitate the interpretation of results from different risk assessments, for instance, for the same LMO.

4. Establishing the context and scope of the risk assessment

When the regulatory process of a country triggers the need for a risk assessment, it usually results in a request from the competent authority to the risk assessor(s). This request includes the scope of the risk assessment to be carried out as well as some important elements that will set the context of the risk assessment. In a typical case-by-case scenario, in accordance with the Cartagena Protocol, these elements will include at minimum: the LMO(s), its(their) specific use(s) and, in cases of introduction into the environment, the likely potential receiving environment(s) where the LMO may be released and establish itself. As such, the case-by-case approach does not allow an existing risk assessment to be applied “as is” to different LMOs, uses or receiving environments. Any request to conduct or review a risk assessment that does not follow the case-by-case principle should lead the regulatory framework to request a new risk assessment with a scope that is specific to the case under consideration (i.e. the LMO, its specific use and the likely potential receiving environment).

Protection goals for the conservation and sustainable use of biodiversity, may be defined in national, regional and international policies. In setting the context of a risk assessment, these goals may be relevant to the identification and selection of appropriate assessment endpoints and to determining which methodology will be used in the risk assessment process. Understanding the contribution of national, regional and regulatory policies in setting the context of the risk assessment is part of the preparatory work for a risk assessment as seen in Module 2.

After consideration of the protection goals, the risk assessment of a particular LMO proceeds to establishing the scope in order to define the extent and the limits of the risk assessment process. This phase usually consists of at least three main actions: (i) selecting relevant assessment endpoints or representative species on which to assess potential adverse effects; (ii) establishing baseline information; and (iii) when possible, establishing the appropriate comparator(s).

Although these actions are described here as separate activities, in practical terms, this is an iterative process where the risk assessors will usually draw on the results of each action to inform the subsequent actions until all their elements have been considered sufficiently enough to enable the risk assessment to proceed.

4.1 Selecting relevant assessment endpoints or representative species

The purpose of an assessment endpoint or of representative species is to provide a measure that will indicate whether or not the LMO may cause an adverse impact on a protection goal. In order to be useful, the selected assessment endpoints or characteristics of the representative species should be specific and measureable.

Assessment endpoints or species representative of important ecological functions or roles should be selected on a case-by-case basis. The complexity of ecosystems and the large number of potential candidates add to the challenges in selecting the appropriate assessment endpoints in ecological systems. Some important criteria for the selection of assessment endpoints to be used in the risk assessment of LMOs may include, for example: (i) their relevance to the protection goals, (ii) a well-defined ecological function, (iii) accessibility to measurement, and (iv) level of potential exposure to the LMO.

Identifying assessment endpoints or representative species that are relevant within the context of the likely potential receiving environment allows the risk assessor(s) to focus on interactions that are likely to occur. Moreover, risk scenarios may be also formulated to include assessment endpoints or representative species that are not present in the likely potential receiving environment but may, nevertheless, be indirectly exposed to the LMOs. This could occur, for example, if a third species, which is sexually compatible with the LMO and the representative species, has a distribution area that overlaps with the distribution areas of the former two providing an indirect exposure pathway between them.

	Example 4 – Common problems in selecting assessment endpoints Endpoint is a goal (e.g. maintain and restore endemic populations); Endpoint is vague (e.g. estuarine integrity instead of abundance and distribution of a species); Ecological entity may not be as sensitive to the stressor; Ecological entity is not exposed to the stressor (e.g. using insectivorous birds for avian risk of pesticide application to seeds); Ecological entities are irrelevant to the assessment (e.g. lake fish in salmon stream); Importance of a species or attributes of an ecosystem are not fully considered; Attribute is not sufficiently sensitive for detecting important effects (e.g. survival compared with recruitment for endangered species). Source: US Environmental Protection Agency (1998).

	Example 5 – Questions asked when selecting representative species for assessing effects of Bt plants on non-target organisms Which variant of the Bt protein are we dealing with? Where is it expressed (in the leaves, pollen or only in the roots)? Is it produced in the plant throughout its life or only during particular growth phases? Which insects come into contact with the Bt protein? Is this contact direct and long-term or only occasional? Which insects ingest the Bt protein through their prey? Source: GMO Safety (website).

4.2 Establishing the baseline

In risk assessment, the baseline information describes the conditions existing prior to the introduction of the factor whose potential adverse effect is being assessed. In simplified terms, the baseline for the risk assessment of an LMO is a snapshot of the environment prior to the introduction of an LMO. Baselines can refer, for instance, to a particular environment or health conditions of a population.

The baseline should be established with the aim of having measurable information on any element of the likely potential receiving environment that is considered relevant in assessing the impacts from the introduction of the LMO, including considerations on possible impacts on human health.

In practice, if relevant assessment endpoints or representative species are selected, the baseline data will consist of data that establishes the conditions of these endpoints or species before the introduction of the LMO in question.

4.3 Establishing the appropriate comparator(s)

As seen above, a comparative approach is one of the general principles of risk assessment as set out in Annex III to the Protocol, where risks associated with the LMO “should be considered in the context of the risks posed by the non-modified recipients or parental organisms in the likely potential receiving environment”.

Using a comparator may help a risk assessor identify the novel characteristics of the LMO and assessing if the LMO presents a greater, lesser or equivalent risk compared to the non-modified recipient organism that is used in a similar way and in the same environment.

The ideal comparator is the closest non-modified genotype to the LMO, i.e. (near-)isogenic lines. Depending on the context, a risk assessor can also choose to consider similar or related non-modified genotypes as useful comparators. Related management practices and experience with similar non-modified organisms may also be helpful. For example, when considering the risk assessment for an insect resistant LM crop, a risk assessor may wish to consider, amongst other things, the available experience with pest control practices applied to non-modified organisms of the same species (e.g. use of spores from Bacillus thuringiensis as pesticides).

In some circumstances, choosing an appropriate comparator(s) could be a challenge. This may happen, for example, in the case of LM crops that are tolerant to abiotic stress if the non-modified recipient or parental organisms are not capable of growing in the receiving environment. In extreme situations, when a suitable comparator cannot be grown under the same conditions and in the same or similar receiving environment as the LMO, it may be necessary to treat the LMO as a novel species in that environment (i.e. introduced species). This means that the characterization of the LMO (see below) will focus not only in the novel genotypic and phenotypic characteristics resulting from the genetic modification, but rather on the characterization of an entire new genotype in the particular receiving environment.

5. Elements of a case-by-case risk assessment of LMOs

The case-by-case approach in risk assessment is based on the premise that risks that may arise from the release of an LMO depend on three main elements: the (i) LMO itself, (ii) the likely potential receiving environment and (iii) the intended use of the LMO in question. In order to identify and assess risks, each of these elements needs to be characterized in a concerted manner and as appropriate for the specific risk assessment. Moreover, it is important to note that while these three elements may be sufficient to establish the boundaries of a risk assessment, potential adverse effects may extend beyond these elements, for instance, to unintended receiving environments and uses.

The information required for each of these elements in a risk assessment may vary in nature and level of detail from case to case. The following sections provide examples of information that may be relevant for the characterization of each element above. These sections include several of the “points to consider” as indicated in paragraph 9 of Annex III of the Protocol.

A large portion of the information listed here is usually included in the LMO request triggering the risk assessment. The risk assessors can determine whether or not the information provided is sufficient and adequate for conducting a scientifically sound risk assessment. If needed, they can obtain additional information by, for instance, carrying out their own investigation or requesting it from the applicant.

	Example 6 – The case-by-case approach “A risk assessment performed for a particular LMO intended to be introduced to one environment may not be sufficient when assessing the possible adverse effects that may arise if that LMO is to be released under different environmental conditions, or into different receiving environments. A risk assessment performed for a particular use of a particular LMO may not be sufficient when assessing the possible adverse effects that may arise if that LMO is to be used in different ways. Because of this, it is important for each case to be addressed separately, taking into account specific information on the LMO concerned, its intended use, and its potential receiving environment.” Source: IUCN (2003).

5.1 Living modified organism

5.1.1 Characterization of the recipient organism

In order to identify whether or not the LMO possesses characteristics that may cause potential adverse effects (see section 5.1), it is first necessary to have information about the non-modified recipient organism (or parental organisms).

For many LMOs, the biology of the recipient organism will strongly influence the potential interactions of the LMO in the receiving environment. Information on the recipient organism is therefore essential as it will help the risk assessor identify the exposure, its scenarios and, ultimately, if any risk is posed by an LMO.

The information that is needed for the characterization of the recipient organism will vary depending on each case. It normally includes the biological and reproductive characteristics of the recipient organism that can be important for determining the potential exposure of other organisms, such as predators, prey, competitors or pathogens, to the LMO in question in the likely potential receiving environment.

For many species of commercialized LMOs, information on the recipient organism can be found in biology documents, such as those published by the Organization for Economic Co-operation and Development (OECD) ³ and the Canadian Food Inspection Agency (CFIA). ⁴

The LMO will, in most cases, share most of its genetic characteristics with its actual recipient organism (i.e. the one used in the modification) rather than with other genotypes of the same species. Thus, it is also important to consider, whenever possible, comparative data from the actual non-modified recipient organism (see the section on “Establishing the appropriate comparator(s)”).

Information about recipient organism to be considered may include:

Taxonomic status – This information is useful for identifying the recipient organism and ensuring that information provided and cited during the assessment pertains to the organism for which the assessment is being carried out. Typically, the taxonomic status includes the scientific name (i.e. genus and species, for example, Zea mays) and information about the taxonomic family (e.g. Poaceae). This may also include other information used to further classify (e.g. sub-species, variety, strain) or differentiate the recipient or parental organism(s) (e.g. ploidy level or chromosome number).

Common name – The familiar or colloquial names for the recipient organism that may be commonly used in the country of introduction and in international trade may be useful for finding information relevant to the biology of the organism. Caution is recommended when using information about recipient organism when only common names (versus the scientific name) are used because the same common name can be applied to more than one species.

Biological characteristics – Information on the biological characteristics of the recipient organism, such as the production of endogenous toxins and allergens, its reproductive biology, seed dispersal and growth habits, are also important points for consideration.

Origin – The origin of the recipient organism refers to its place of collection and may be important because populations within a species (e.g., variety, strain, isoline, etc.) may have significantly different characteristics. For domesticated species, this may be supplemented with a pedigree map where available.

Centres of origin and centres of genetic diversity – Knowledge of the centre(s) of origin and genetic diversity can provide information on the presence of sexually compatible species and the likelihood of ecological interactions in the receiving environment. In the absence of more specific information, the centre of origin can also offer insight into the biology of the species (e.g. habitats to which the species is adapted).

Habitat where the recipient or parental organism(s) may persist or proliferate – Information about the ecosystems and habitats (e.g. temperature, humidity, altitude, etc) where the recipient organism is known to be native and where it may have been introduced and is now established provides useful baseline information. This allows the risk assessors to understand the range of habitats in which the species exists, the range of behaviours exhibited in those habitats, and how characteristics of the species determine the range of habitats where it can persist or proliferate. This information can be very valuable in determining the likely potential receiving environment and, consequently, the level of exposure to the LMO. Likewise, the ecological characteristics of the recipient organism will help determine which organisms in the likely potential receiving environment are likely to come into contact, either directly or indirectly, with the LMO and will help determine the exposure pathways. For more details on the type of information that may be useful, see section 4.2 on the “Likely potential receiving environment”.

The history of use can be very valuable as well. If an organism persists in heavily managed environments (e.g. agriculture, silviculture or recreationally managed land) then this will provide information about the conditions necessary for its survival. It may also provide direct indications of how the LMO will behave in other managed environments.

5.1.2 Description of the genetic modification

Information on the genetic material that was introduced or modified, as well as the method used for the genetic transformation is useful in identifying novel properties of the LMO such as, what new gene products are expressed and which of the endogenous genes of the recipient or parental organism(s) may be affected by the genetic modification.

Typically the description of the genetic modification includes information on (i) the “donor organism(s)” or the source of the inserted genetic element(s); (ii) characteristics of each modified genetic element, including their intended and known biological function(s); (iii) the vector used, if applicable; and (iv) the transformation method. Below is a brief explanation on each of these points:

Donor organism(s) – The relevant information on the donor organism(s) includes its taxonomic status, common name, origin and relevant biological characteristics.

Modified genetic elements – The relevant information on the modified genetic elements encompasses the name, sequence, function and other characteristics of all the nucleic acid sequences that were modified in the LMO. These include not only the target gene but also, for example, all marker genes, regulatory sequences, and any non-coding DNA. If available, a history of use may be important with regards to potential toxicity or allergenicity of the gene products derived from the donor organism. If the genetic elements originate from a donor organism that is known to be a pest or pathogen it is also relevant to know if and how these elements contribute to the pest or pathogenic characteristics.

Vector – In molecular biology, a vector is a nucleic acid molecule used as a vehicle to transfer foreign genetic material into another cell. If a vector, for example a plasmid, was used for the transformation, relevant information includes its identity, source or origin, and its host range.

Transformation method – Specifying the method that was used in the transformation (e.g. Agrobacterium mediated, particle gun, etc.) is also relevant when describing the genetic modification. Depending on the transformation method, parts of the vector(s) may also be incorporated into the genome of the newly developed LMO.

Characteristics of the modification – This refers to information about whether or not the inserted or modified genetic elements are present and functioning as expected in the LMO. Normally this involves confirmation that the DNA insert or modified genetic element is stable in the genome of the LMO. Information such as the insertion site in the genome of the recipient or parental organism(s), cellular location of the insert (e.g. chromosomal, extrachromosomal, or chloroplast DNA), its mode of inheritance and copy number may also be relevant.

5.1.3 Identification of the LMO

With regard to the identification of the LMO, the following are important points to consider:

Unique identifiers – A Unique identifier is a code provided by the LMO developer to a transformation event derived from recombinant DNA techniques to enable its unequivocal identification. Each unique identifier is made up of a sequence of 9 alphanumeric digits, for example MON-89788-1, assigned according to the OECD guidance document (OECD, 2006).

Detection and identification methods – The availability of methods for detection and identification of the LMO may be considered as well as their specificity, sensitivity and reliability. This information may be relevant not only for assessing risks but also when considering possible monitoring and risk management strategies (see Module 4). Some regulatory frameworks require a description of such methods as a condition for regulatory approval in order to ensure the tools to assist with monitoring and risk management are available.

The Biosafety Clearing-House of the Cartagena Protocol maintains an LMO registry containing, amongst other things, information on unique identifiers, molecular characteristics and available detection methods for the LMOs addressed in countries’ decisions.

	Example 7 – CFIA detection and identification method criteria According to the Canadian Food Inspection Agency (CFIA), acceptable methods for detection and identification of LMOs must address the following: Test type – Methods must be suitable and may be protein, RNA- or DNA-based. Phenotypic-based methods will not generally be considered suitable. Limit of detection – Methods must meet the following sensitivity and accuracy requirement: For those methods that are grain-based, the method must be able to detect 0.2% modified grain (2 grains in 1000) with a 95% confidence interval. For those methods that are not grain-based (e.g. single ingredient feed), the method must be able to detect 0.2% modified material in a sample with a 95% confidence interval. Procedural clarity – The method must be complete and laid out in a stepwise fashion that may be easily followed by a person unfamiliar with the method. Detailed descriptions of sample size, replicates, extraction procedure, expected results (figures/sequences), interpretation and acceptance criteria must be included. Cross-reactivity – The method must be shown to be specific to the PNT of interest. Any potential for cross-reactivity must be clearly stated. Cross-reactivity data must be provided demonstrating that the method does not cross-react with other commercially available PNTs of the same species with similar traits/modifications that are currently available on the Canadian market. Reference material – The company must provide appropriate reference materials to the CFIA upon request. Appropriate reference material will be determined by the CFIA based on the method provided. Contact information – The company must provide contact information for a technical support person. Source: CFIA (website).

5.2 Likely potential receiving environment(s)

The Protocol calls for the characterisation of the “likely potential receiving environment” of an LMO. This encompasses both the area where the LMO will be intentionally introduced into the environment as well as any other environment which may be exposed to the LMO. As such, during a risk assessment in accordance with the Protocol, in addition to the area where the LMO will be intentionally introduced, the likely potential receiving environment of an LMO should also be thoroughly examined with particular attention given to areas where exposure levels to the LMO will be the highest.

The characterization of the likely potential receiving environment takes into account its ecological characteristics, including physical location/geography, climate, its biological entities and their interactions. The characterization of the likely potential receiving environment will help in selecting appropriate assessment endpoints for the risk assessment (see Module 2, section 2.1) and will also affect the assessment of the potential interactions of the LMO with other organisms.

To determine the likely potential receiving environment, risk assessors may consider potential pathways for dispersal of the LMO as well as the habitats where the recipient/parent organism(s) may persist or proliferate.

An analysis of possible dispersal routes and mechanisms is important when establishing the likely potential receiving environments. Different dispersal mechanisms may exist and could be inherent either to the LMO (e.g. altered seed characteristics), its intended use (e.g. shipment practices) or the receiving environment (e.g. proximity to a river). The risk assessment should take into consideration all possible dispersal mechanisms, keeping in mind the biology of the LMO and non-modified recipient or parental organism(s), in a concerted manner for each case.

Information about the likely potential receiving environment can include considerations on both large scale (e.g. climate) and small scale characteristics (e.g. microclimate) depending on the complexity of the environment. The type of information on the likely potential receiving environment and the level of detail depend on the nature of the LMO and its intended use, in accordance with the case-by-case principle. (see section 4.3).

It may not be possible or practical to consider every possible interaction between the LMO and the receiving environment. Such challenges and limitations should be acknowledged during the risk assessment process. Below are descriptions of some physical and biological characteristics of the likely potential receiving environment(s) that can be considered in the risk assessment of LMOs. This is an indicative list thus the information required to satisfy the needs of the assessment will vary depending on the nature of the LMO and its intended use.

5.2.1 Physical characteristics

The physical or “abiotic” characteristics of the likely potential receiving environment may have a great impact on the ability of an LMO to survive and persist.

Geography and climate – Geography encompasses characteristics such as latitude, which will influence day-length, and altitude. Climate encompasses temperature, precipitation, humidity, wind and other meteorological measures over long periods of time. For the purposes of environmental risk assessment, geography and climate are among the most important factors impacting the ability of an LMO to survive and persist. For LM plants, temperature and precipitation are likely to be key determinants. Seasonality (variations in climate on an annual cycle) can also be an important consideration in the potential survival and persistence of an LMO.

Soil – The type and quality of soil can greatly influence the ability of an LM plant to survive or persist without land management. The type and quality of a soil are heavily influenced by the organisms living in its proximity, but abiotic factors such as climate, geography and topography will also all play a role in determining its characteristics (e.g. mineral content, moisture level, texture etc.).

Management status – The management status of an environment is a measure of how much human intervention takes place in order to maintain a particular condition. A separate but related concept is “disturbance” which can be considered the amount of human activity that affects the environment but without the intention of maintaining a particular condition. Management and disturbance may greatly influence the ability of an LMO to survive and persist in the environment. Likely potential receiving environments can range from highly managed to unmanaged and from highly disturbed to undisturbed.

5.2.2 Biological characteristics

The biological characteristics of the likely potential receiving environment consist of all the living organisms present in the environment, its biological communities and the interactions among them.

Both managed and unmanaged environments contain complex biological characteristics that pose challenges for environmental risk assessments.

As with any other organism, an LMO released into the environment is expected to have many interactions with other organisms. For the purposes of environmental risk assessment, it is critical to develop verifiable risk scenarios and identify the appropriate species that may be impacted by the presence of the LMO in the environment. For example, gene flow and possibly introgression are more likely to occur when sexually compatible species are present in the likely potential receiving environment. The selection of suitable representative species in the likely potential receiving environment is also informative (see section 3.1).

5.3 Intended use

The characteristics of the intended use of an LMO and management practices associated with it, such as tilling and the use of pesticides, can provide valuable information and context for the risk assessment process. Understanding the intended use also helps a risk assessor to perform an exposure assessment starting with the environment where the LMO will be deliberately introduced followed by considering whether or not the LMO is likely to disseminate or persist outside of this environment.

To illustrate how the intended use can affect the likelihood of a risk posed by an LMO, a hypothetical case of an LM tree being used for wood production could be considered, in which the first flowering would occur after 15 years of planting, but logging would takes place after only 10 years. As such, the intended use would result in the LM tree being logged before its first flowering. Consequently, in this hypothetical case, the intended use would influence the likelihood of potential outcrossing of this LM tree.

Information regarding the intended use of the LMO may also take into account any new or changed use in comparison to the recipient or parental organism(s), for example, in cases where the recipient or parental organism(s) is a crop for human consumption but the intended use of the LMO is the production of a compound for pharmaceutical or industrial use.

The scale and type of the introduction into the environment, for example, field trials versus commercial releases, and whether or not any risk management strategy is being proposed, may also be relevant when considering the intended use. Many regulatory frameworks, for instance, require that submissions for field trials be accompanied by information on risk management strategies to reduce exposure to the LMO.

Considerations on the intended use may also take into account national and regional experiences with similar organisms, their management and exposure to the environment.

6. Conducting the risk assessment

Conducting the risk assessment involves synthesizing what is known about the LMO, its intended use and the likely potential receiving environment to establish the likelihood and consequences of potential adverse effects to biodiversity and human health resulting from the introduction of an LMO. Risk assessors need to identify the information needed and understand how it will be used. Using and interpreting existing information, as well as identifying information gaps and understanding how to deal with scientific uncertainty are crucial actions during the risk assessment.

Some risks can be assessed based on existing scientific literature and previously available information alone. Others may require laboratory experiments (e.g. early tier toxicology testing), confined field experiments or other scientific observations. Scientifically sound methodologies should be determined and documented for testing any identified risk scenario. When assessment methods are well described, risk assessors and subsequent reviewers are better equipped to determine whether the information used was adequate and sufficient for characterizing the risk.

	Example 8 – Data acquisition, verification and monitoring “The importance of the data acquisition, verification, and monitoring process in the development of accurate risk assessments has been emphasized. Models, no matter how sophisticated, are simply attempts to understand processes and codify relationships. Only the reiteration of the predictive (risk assessment) and experimental (data acquisition, verification, and monitoring) process can bring models close to being a true picture of reality.” Source: UNEP/IPCS (1994).

Considerations of uncertainty are undertaken throughout the whole risk assessment process. The risk assessment methodology as set out by the Cartagena Protocol states that “where there is uncertainty regarding the level of risk, it may be addressed by requesting further information on the specific issues of concern or by implementing appropriate risk management strategies and/or monitoring the living modified organism in the receiving environment”. ⁵

Although uncertainty can be often addressed by requesting additional information, the necessary information may not always be available or new uncertainties may arise as a result of the provision of additional experimental data. Uncertainty is inherent to the concept of risk thus it is important to consider and analyze, in a systematic way, the various forms of uncertainty (e.g. types and sources) that can arise at each step of the risk assessment process.

Uncertainties may arise from: (i) lack of information, (ii) incomplete knowledge, and (iii) biological or experimental variability, for example, due to inherent heterogeneity in the population being studied or to variations in the analytical assays. Uncertainty resulting from lack of information includes, for example, information that is missing and data that is imprecise or inaccurate (e.g., due to study designs, model systems and analytical methods used to generate, evaluate and analyze the information) (SCBD, 2012).

If the level of uncertainty changes during the risk assessment process (e.g. by provision of new information), an iteration of parts or the entire risk assessment process may be needed.

It is important to note that while scientific uncertainty is considered during the risk assessment process and the results of uncertainty considerations may be reported; ultimately it is the responsibility of the decision-makers to decide how to use the information in conjunction with the principals of the precautionary approach when making a decision on an LMO.

	Example 9 – Scientific uncertainty “There is no internationally agreed definition of ‘scientific uncertainty’, nor are there internationally agreed general rules or guidelines to determine its occurrence. Those matters are thus dealt with – sometimes differently – in each international instrument incorporating precautionary measures.” Source: IUCN (2003).

The following sections will address the steps of the risk assessment methodology described in paragraph 8 of Annex III to the Protocol.

These steps describe a structured and integrated process whereby the results of one step are relevant to subsequent steps. Additionally, the risk assessment process may need to be conducted in an iterative manner, whereby certain steps may be repeated or re-examined to increase or re-evaluate the reliability of the risk assessment. If during the process, new information arises that could change the outcome of a step, the risk assessment may need to be re-examined accordingly.

6.1 Identification of any novel genotypic and phenotypic characteristics associated with the LMO that may have adverse effects

The first step of the risk assessment is “an identification of any novel genotypic and phenotypic characteristics associated with the LMO that may have adverse effects on biological diversity in the likely potential receiving environment, taking into account risks to human health”. ⁶

What constitutes an “adverse effect” will depend on the context and scope of the risk assessment taking into account, as appropriate, the specific protection goals as seen above.

	Example 10 – Potential adverse effects “With every new emerging technology, there are potential risks. For LMOs, the potential risks include: The danger of unintentionally introducing allergens and other anti-nutrition factors in foods; The likelihood of transgenes escaping from cultivated GM crops into wild relatives; The potential for pests to evolve resistance to the toxins produced by GM crops; The risk of these toxins affecting non-target organisms.” Source: GMAC Singapore (website).

The molecular and phenotypic characterization of an LMO provides the basis for identifying differences, both intended and unintended, between the LMO and its recipient or parental organism(s). Molecular analyses may be performed to characterize the products of the modified genetic elements, as well as of other genes that may have been affected by the modification. Data on specific expression patterns may be relevant for risk assessment in order to determine exposure, and may also include data confirming the absence of unintended products (e.g. in the case, for instance, where the gene product is intended to function only in a specific tissue, data may be used to confirm its specificity in that tissue and demonstrate its absence in other tissues).

Other phenotypic data are often presented to indicate that the LMO is behaving as anticipated. This could include data on reproductive characteristics, alterations in susceptibility to pests and diseases or tolerance to abiotic stressors, etc.

Once the potential adverse effects have been identified, the risk assessment proceeds to estimating the likelihood and consequences of these effects. To this end, developing risk scenarios may in some cases provide a useful tool.

A risk scenario may be defined as a theoretical sequence of events with an associated probability and consequence. In the context of risk assessment of LMOs, a risk scenario may be explained as a scientifically supportable chain of events through which the LMO might have an adverse effect on an assessment endpoint.

	Example 11 – A risk scenario “The possibility that growing Bt corn may kill ladybird beetles due to ingestion of the Bt protein when preying on insects feeding on the GM corn, thereby reducing the abundance of coccinellids in the agroecosystem and increasing the incidence of pests.” Source: Hokanson and Quemada (2009).

A well defined risk scenario should be scientifically plausible and allow the assessor to identify information that is necessary for the assessment of risks.

Although some risk scenarios may appear as obvious (e.g. potential for insect resistant plants to affect insect herbivore populations), it is always useful to identify the risk scenarios fully.

Clear and well-defined risk scenarios can also contribute to the transparency of a risk assessment because they allow others to consider whether or not the subsequent steps of the risk assessment have been adequately performed and facilitate the consideration of possible strategies to manage the identified risks.

A common challenge in generating a well-defined risk scenario is to choose representative species that would be exposed to the LMO. This is why an exposure assessment should be considered when selecting assessment endpoints.

When establishing risk scenarios several considerations may be taken into account. These may include: (i) gene flow followed by undesired introgression of the transgene in species of interest; (ii) toxicity to non-target organisms; (iii) allergenicity; (iv) tri-trophic interactions and indirect effects; and (v) resistance development. The following paragraphs explain some of these considerations in more detail:

Gene flow followed by undesired introgression of the transgene in species of interest – Gene flow is a term used to indicate the transfer of genetic material from one population or species to another. Gene flow may be horizontal (i.e. without involvement of sexual crossing) or vertical (e.g. seed production via pollen).

In the case of plants, vertical gene flow may occur even between organisms that are located far apart since pollen can be carried across large distances by the wind or insects, for instance.

The potential for gene flow from an LMO to non-modified organisms is first evaluated by investigating if sexually compatible species are present in the likely potential receiving environment. If sexually compatible species are present, there is a possibility of gene flow from the LMO to these species. Whether or not the modified genetic elements can potentially introgress into the population of the sexually compatible species will be largely determined by the biology of the recipient organism and of the LMO itself (see considerations regarding the likelihood and consequences of gene flow and introgression in sections 5.2 and 5.3).

	Figure 4 - Gene flow to conventional crops and distant relatives through “genetic bridges” Source: Heinemann (2007).

Toxicity to non-target organisms – The potential for an introduced gene product to be toxic to organisms in the environment is typically addressed by controlled exposure in the environment or by direct toxicity testing, or by a combination of the two. Non-target organisms may include, for instance, herbivores, natural enemies (e.g. parasitoids and predators), pollinators and pollen feeders, soil ecosystems and weeds.

If toxicity testing is needed, it typically follows a sequential series of tiered tests. Early tier studies involve highly controlled laboratory environments where representative or surrogate test species are exposed to high concentrations of the gene product being studied (i.e. worst case exposures) to determine if there are any toxic effects. If toxic effects are observed in early tier tests or if unacceptable uncertainty exists, more realistic conditions representative of field-level exposures can be tested to determine the extent of the risk.

The gene products of the modified genetic elements in LMOs may be produced in very small quantities thus may be difficult to isolate in the amounts required for toxicity testing. If this is the case, and it is determined that toxicity tests are required, the risk assessor may consider results from tests using gene products obtained from alternate (surrogate) sources (e.g. bacterial expression systems or the organism from which the transgene was derived) provided that these gene products are chemically and functionally equivalent.

	Figure 5 – Exposure to non-target organisms Source: VADLO (website).

Allergenicity – Allergies are a type of adverse immunological response that affect individuals who are predisposed to certain types of substances (i.e. allergens). Allergens are often proteins or peptides.

In considering allergenicity caused by LMOs, it is important to take into account the exposure to proteins newly expressed by the LMO, including some variants of these proteins that may be produced uniquely by the LMO. As a consequence, some allergenicity studies must be carried out with proteins isolated from the LMO itself, and not obtained from alternate (surrogate) source such as a bacterial expression system).

It is also possible that allergens known to exist in the recipient or parental organism(s) are produced in higher amounts, for example by over-expression of the gene that encodes a protein that is known to be a common allergen.

	Example 12 – Assessment of the allergenic potential of foods derived from modern biotechnology Source: FAO/WHO (2001).

Tri-trophic interactions and indirect effects – “Tri-trophic interaction” is an important concept in ecology and occurs when a change at one trophic level indirectly affects trophic levels which are more than one step away. Consideration of tri-trophic interactions and indirect effects may be relevant to biodiversity protection goals.

	Example 13 – A tri-trophic interaction “Suppose that there were a grassland where the major herbivore was a species of vole (n.b. a small rodent) which eats grass seeds and that this vole was able to reach population levels which allowed the vole to eat nearly all of the seeds. Further suppose that the main predator of this vole was a species of hawk and that this hawk was capable eating enough voles to reduce the voles population to nearly zero (at least to the point that voles could no longer eat very many of the seeds). So, if the population of hawks is high, the population of voles is low and the grass produces lots of seeds. However, if the population of hawks is low, the vole population will be high, and the grass will disperse few seeds.” Source: Abrahamson (website).

Observations and experimentation to identify such effects are challenging because of the complexity of ecological interactions, the difficulty of establishing causality between observed variation and treatment effects (e.g. the presence of the modified genetic element or its products), and natural variability in populations over time. Moreover, in a food chain (or food web), effects at the trophic levels may become observable only at a later stage.

Resistance development – The extensive use of herbicides and insect resistant LM crops has the potential to result in the emergence of resistant weeds and insects. Similar breakdowns have routinely occurred with conventional crops and pesticides. Several weed species have developed resistance to specific herbicides which are extensively used in combination with herbicide-resistant LM crops. Insect-resistant Bt-crops similarly could lead to the emergence of Bt-resistant insects (FAO, 2004).

The extent of the adverse effect and possible consequences of the insurgence of resistant weeds and insects should be thoroughly considered in a risk assessment. Some regulatory frameworks require that risk management strategies are identified in order lower the risk of resistance development.

	Example 14 – Topics of concern According to the International Centre for Genetic Engineering and Biotechnology (ICGEB), the main issues of concern derived from the deliberate introduction of LM crops (and their derived products) into the environment or onto the market have been classified as: Risks to animal and human health – Toxicity and food/feed quality/safety; allergies; pathogen drug resistance (antibiotic resistance); impact of selectable marker; Risks to the environment – Persistence of gene or transgene (volunteers, increased fitness of LM crop, invasiveness) or of transgene products (cumulative effects); susceptibility of non-target organisms; change in use of chemicals in agriculture; unpredictable gene expression or transgene instability (gene silencing); environmentally-induced (abiotic) changes in transgene expression; ecological fitness; changes to biodiversity (interference of tri-trophic interactions); impact on soil fertility/soil degradation of organic material; Gene transfer – Genetic pollution through pollen or seed dispersal and horizontal gene transfer (transgene or promoter dispersion); transfer of foreign gene to microorganisms (DNA uptake); or generation of new live viruses by recombination (transcapsidation, complementation, etc.); Risks for agriculture – Resistance/tolerance of target organisms; weeds or superweeds; alteration of nutritional value (attractiveness of the organism to pests); change in cost of agriculture; pest/weed management; unpredictable variation in active product availability; loss of familiarity/changes in agricultural practice. Source: ICGEB (website).

6.2 Evaluation of the likelihood

This step entails an evaluation of the likelihood of adverse effects being realized, taking into account the level and kind of exposure to the LMO by the likely potential receiving environment.

After the potential adverse effects of the LMO have been identified, the risk assessment proceeds to a formal analysis of the likelihood and consequence of these effects with respect to the identified assessment endpoints.

Although the steps of evaluating likelihood and consequences are dealt with separately in Annex III of the Protocol, some risk assessment approaches consider these steps simultaneously or in reverse order.

The likelihood of an adverse effect is dependent upon the probability of one or a series of circumstances actually occurring. It is difficult to describe in detail an evaluation of likelihood or consequence without using an example because the evaluation is dependent on the nature of the LMO, the receiving environment and, if appropriate, on the risk scenario used. The following are two examples:

• In a case where undesired outcrossing of the transgene with a non-modified organism is determined to be possible (i.e. the two species are sexually compatible), the risk assessment may consider both the likelihood of the outcrossing and, if relevant, the likelihood of the LMO progeny to persist or proliferate. Considerations on the latter may be based, for example, on assessing whether or not the transgene would affect the fitness level of the progeny (i.e. the capability of individuals to compete and reproduce in a given environment). If the transgene induces a positive fitness effect, the likelihood that the population resulting from the outcrossing would increase is high. On the other hand, transgenes that have a negative fitness effect would result in a low likelihood that the resulting population would increase. Transgenes that have a neutral impact on fitness may persist in populations at low levels depending on the rate of outcrossing or introgression as well as the overall population dynamics of the species.
• In a case where the risk scenario involves the potential toxicity of an LMO plant (or a substance produced by an LMO plant) to a herbivorous insect: the analysis of likelihood may consider the probability that the insect will be present, that the insect will feed on the LMO and that the insect will ingest a sufficient quantity of the LMO to suffer an adverse effect. Likelihood may consider probabilities on an individual level (e.g. what are the chances an individual insect may consume the LMO plant) or on a population level (e.g. what percentage of the population of insects will come into contact with the LMO) or both.

	Example 15 – Likelihood of introgression “To evaluate a possible ecological effect of an inserted gene being introgressed into a natural population it is important to estimate the probability of introgression. Such a probability estimate can be obtained from measurements of hybridisation rates, assumed selective advantage of inserted gene, and fitness measurements of parent plants, hybrid plants, and plants from the first and second back-cross generations. If hybrids are formed and it is likely that these hybrids are able to survive the consequences should be discussed.” Source: Ministry of Environment and Energy Denmark (1999).

6.3 Evaluation of the consequences

The consequences of the adverse effects, should these occur, may be severe, minimal, or anywhere in between. The evaluation of the consequences may consider the effects on individuals (e.g. mortality, reduced or enhanced fitness, etc.) or on populations (e.g. increase or decrease in number, change in demographics, etc.) depending on the adverse effect under evaluation.

The risk assessment should consider the consequences of each adverse effect based on a concerted analysis of what is known about the LMO, the likely potential receiving environment and the assessment endpoints, as well as the likelihood assessment.

	Example 16 – Consequences of effects to non-target organisms “When the inserted trait causes the plant to produce potentially toxic compounds, or if flower characteristics are changed, i.e. colour, flowering period, pollen production, etc., then effects on pollinators have to be measured. A test of effects on honeybees (Apis mellifera) is obligatory because of the importance of honeybees as pollinators of both wild and crop species and because standardized test protocols testing for effects of conventional pesticides exist for this pollinator. These tests include exposure through nectar and pollen.” Source: Ministry of Environment and Energy Denmark (1999).

Also using an example where gene flow and introgression could lead to a potential adverse effect, what impact the presence of a transgene will have on biodiversity will depend on its effect on individual fitness as well as on the importance of that species relative to the protection goals. For instance, if a sexually compatible species, present in the receiving environment, is directly relevant to a biodiversity protection goal (e.g. it is a protected species) then the impact on biodiversity can be assessed by looking directly at the impact of the transgene on the population. If the sexually compatible species is not directly related to a biodiversity management goal, then the impact of the expression of the transgene will be dependent on indirect interactions. Indirect effects may be challenging to assess (see section 5.1), and are dependent on the ecological importance of the species.

6.4 Estimation of the overall risk

This step consists of the integration of the likelihood and consequence of each of the individual risks identified through the preceding steps and takes into account any relevant uncertainty that emerged thus far during the process. In some risk assessment approaches, this step is referred to as “risk characterization”.

To date, there is no universally accepted method to estimate the overall risk but a variety of guidance materials are available that address this topic (see for instance, documents under “Scientific and technical issues / risk assessment” in the Biosafety Information Resource Centre, BIRC). ⁷

In rare instances, the risk characterization results in a quantitative value (e.g. 6% of a population will be exposed to a stressor, and of that percentage half will experience mortality). More frequently, the risk characterization for an LMO will be qualitative. In such cases, description of the risk characterization may be expressed as, for instance, ‘high’, ‘medium’, ‘low’, ‘negligible’ or ‘indeterminate due to uncertainty or lack of knowledge’.

The outcome of this step is the assessment of the overall risk of the LMO. Once this is achieved, it is helpful to determine, as an internal quality control, whether the risk assessment has met the criteria established at the beginning of the process taking into account also those criteria established in the relevant policies in practice with regard to the protection goals, assessment endpoints and thresholds.

	Figure 6 – Estimation of overall risk Source: ERMA NZ (1998).

6.5 Identification of risk management and monitoring strategies

Annex III of the Protocol states that the risk assessment methodology may entail “a recommendation as to whether or not the risks are acceptable or manageable, including, where necessary, identification of strategies to manage these risks” and “where there is uncertainty regarding the level of risk, it may be addressed by requesting further information on the specific issues of concern or by implementing appropriate risk management strategies and/or monitoring the living modified organism in the receiving environment”. ⁸

6.5.1 Risk management

Risk management strategies refer to measures that may be implemented after the LMO is introduced into the environment (or placed in the market, if applicable) aimed at reducing the risks identified during the assessment to a level that may be considered as acceptable. Risk management strategies can be useful to increase confidence when dealing with uncertainty or, in the case where risks have been identified, to reduce the likelihood or impact of the potential adverse effect.

	Example 17 – Application of management strategies for risks from the deliberate release or marketing of LMO(s) “The risk assessment may identify risks that require management and how best to manage them, and a risk management strategy should be defined.” Source: The European Parliament and the Council of the European Union (2001).

Risk management strategies may aim to reduce the likelihood or consequences of potential adverse effects and are referred to as “preventive measures” and “mitigation measures”, respectively. Some approaches to risk assessment may also include the identification of measures to control an adverse effect should it occur.

For LMOs, common risk management strategies have typically been designed to reduce the likelihood of exposure, but depending on the specific case, management options might include a variety of measures that are directly or indirectly related to the LMO. Some examples of risk management strategies for LMOs include: minimum distances from sexually compatible species if there is evidence that gene flow could cause adverse effects, destruction of seeds remaining in the field or of volunteer after harvest, restrictions from introduction into specified receiving environments, etc.

Certain risk assessment steps, particularly the evaluation of likelihood and consequences may need to be re-evaluated to take into account each of the identified risk management strategies since these may affect the estimation of the overall risks.

6.5.2 Monitoring

A risk assessor may identify the need for a strategy to monitor the receiving environment for adverse effects that may arise after the introduction of the LMO and include it as part of the recommendations for the Competent National Authority(ies). This may happen, for instance, when the level of uncertainty could affect the overall conclusions of the risk assessment. Moreover, some biosafety frameworks may have a policy to request a plan for monitoring as part of the risk assessment of all or particular types of LMOs.

Monitoring after the release of the LMO aims at detecting changes (e.g. in the receiving environment(s) or in the LMO) that could lead to adverse effects.

	Example 18 – Post-market monitoring “Post-market monitoring may be an appropriate risk management measure in specific circumstances. Following the safety assessment, the need and utility for post-market monitoring should be considered, on a case-by-case basis, during risk assessment and its practicability should be considered during risk management.” Source: Health Canada (2006).

Monitoring strategies may be designed on the basis of the protection goals identified by national legislation and regulation, if available, and parameters that are relevant to the indication of any increasing risk to the assessment endpoints in a “top-down” approach, or on the basis of specific risks in a “bottom-up” approach.

The strategies may include “general surveillance”, designed to identify unexpected effects of the LMOs or traits, such as long-term effects; or be “case-specific” where potential adverse effects identified during the risk assessment are investigated. Monitoring for the development of resistance in insect pests following introduction of pesticide producing LM crops would be an example of a “case-specific” scenario. Monitoring for the abundance of beneficial insect species in an environment would be an example of “general surveillance”.

	Example 19 – Case-specific monitoring and general surveillance of LM plants “The environmental monitoring of the GM plant will have two focuses: (1) the possible effects of the GM plant, identified in the formal risk assessment procedure, and (2) to identify the occurrence of adverse unanticipated effects of the GM plant or its use which were not anticipated in the environmental risk assessment. Appropriate case-specific monitoring measures should be developed on a case-by-case approach depending upon the outcomes of the risk assessment. Possible risks identified in the environmental risk assessment should be studied in hypothesis-driven experiments and tests. The objective of general surveillance is to identify the occurrence of unanticipated adverse effects of GM plants or their use on human health or the environment that were not anticipated in the environmental risk assessment. Since no specific risk is identified, no hypothesis of risk can be tested, so it is difficult to propose specific methods to carry out general surveillance.” Source: EFSA (2006).

Where it is appropriate, other potential adverse effects such as delayed, cumulative, synergistic or indirect effects resulting from the LMO, the trait or the inserted or modified genes may be considered in the post-release monitoring strategies.

The level of specificity of the risk management and monitoring strategies may vary depending on the LMO(s), the intended use(s) and/or the likely potential receiving environment(s). Therefore, it is essential that a detailed methodology for each identified strategy also be identified. The methodology may include, for example, the frequency, locations and methods of sampling, as well as methods of analysis (e.g. laboratory testing).

	Example 20 – Various types of monitoring according to the Australian Government Routine monitoring inspections – these are based on risk profiling and sampling of a range of dealings, locations where dealings are undertaken, and organisations who are conducting dealings; Follow-up visits – these are undertaken to follow-up on issues or to check the implementation of remedial action; Review visits – monitoring of premises may be focused on a specific issue that is being reviewed by the Monitoring and Compliance Sections and visits are selected on that basis; Audit visits – a comprehensive examination of an organisation’s activities that includes specific visits to inform the audit process; Investigation visits – these visits are based on inquiries into allegations of a breach of the Gene Technology Act 2000; and Unannounced ‘spot checks’ – these are undertaken as a subset of the routine monitoring activities or as part of follow-up checks, incident reviews, or investigations. Source: OGTR (2007).

7. References

1. Abrahamson WG (website) The Solidago Eurosta Gall Homepage – A Resource for Teaching and Research. Ecology and Evolution. Available at http://www.facstaff.bucknell.edu/abrahmsn/solidago/gallresearch.html (access June 2010).

2. CFIA (website) Detection and Identification Method Criteria. Canadian Food Inspection Agency (CFIA). Available at http://www.inspection.gc.ca/english/plaveg/bio/detecte.shtml (access May 2010).

3. EEA (1998) Environmental Risk Assessment - Approaches, Experiences and Information Sources. Environmental issue report No 4, European Environmental Agency (EEA). Available at http://www.eea.europa.eu/publications/GH-07-97-595-EN-C2/riskindex.html (access July 2010).

4. EFSA (2006) Opinion of the Scientific Panel on Genetically Modified Organisms on the Post Market Environmental Monitoring (PMEM) of genetically modified plants (Question No EFSA-Q-2004-061). European Food Safety Authority (EFSA). Available at http://www.efsa.europa.eu/en/efsajournal/doc/gmo_op_ej319_pmem_en,0.pdf (access June 2010).

5. ERMA NZ (1998) Annotated methodology for the consideration of applications for hazardous substances and new organisms under the HSNO Act 1996. Environmental Risk Management Authority of New Zealand (ERMA NZ), 30 pp. Available at http://www.epa.govt.nz/Publications/Annotated-Methodology.pdf (access August 2012).

6. FAO (2004) The State of Food and Agriculture: 2003-2004. Part I: Agricultural biotechnology: meeting the needs of the poor? Section B: The evidence so far. Food and Agriculture Organization of the United Nations. Available at http://www.fao.org/docrep/006/Y5160E/y5160e00.htm (access May 2010).

7. FAO (2011) Biosafety Resource Book. Food and Agriculture Organization of the United Nations (FAO), Module C, 90pp. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=102000 (access September 2011).

8. FAO/WHO (2001) Evaluation of Allergenicity of Genetically Modified Foods. Food and Agriculture Organization of the United Nations (FAO) and World Health Organization (WHO), 29 pages. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=41976 (access June 2010).

9. GMAC Singapore (website) Genetically Modified Organisms. Genetic Modification Advisory Committee (GMAC) Singapore. Available at http://www.gmac.gov.sg/Index_FAQs_Genetically_Modified_Organisms.html (access August 2010).

10. Health Canada (2006) Guidelines for the Safety Assessment of Novel Foods. Food Directorate Health Products and Food Branch, Health Canada, June, 2006. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=101221 (access June 2010).

11. Heinemann JA (2007) A typology of the effects of (trans)gene flow on the conservation and sustainable use of genetic resources. Food and Agriculture Organization (FAO), Background Study Paper no. 35 rev. 1, 100 pp. Available at http://bch.cbd.int/database/record.shtml?documentid=48542 (access August 2012).

12. Hill RA (2005) Conceptualizing risk assessment methodology for genetically modified organisms. Environ. Biosafety Res. 4: 67-70. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=41660 (access June 2010).

13. Hokanson K, Quemada H (2009) Improving risk assessment – problem formulation and tiered testing. Presented at SEARCA Agriculture and Development Seminar Series, 28 April 2009. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=101212 (access June 2010).

14. ICGEB (website) International Centre for Genetic Engineering and Biotechnology. Available at http://www.icgeb.org/~bsafesrv/introduction/generalintro.html (access May 2010).

15. IUCN (2003) An Explanatory Guide to the Cartagena Protocol on Biosafety. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=41476 (access June 2010).

16. Ministry of Environment and Energy Denmark (1999) Ecological Risk Assessment of Genetically Modified Higher Plants (GMHP) – Identification of Data Needs. Ministry of Environment and Energy, National Environmental Research Institute (NERI) Denmark. NERI Technical Report, No. 303, 35 pp. Available at http://www2.dmu.dk/1_viden/2_Publikationer/3_fagrapporter/rapporter/fr303.pdf (access July 2010).

17. OECD (2006) Guidance for the Designation of a Unique Identifier for Transgenic Plants. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=101186 (access June 2010).

18. OGTR (2007) Monitoring protocol in accordance with the Gene Technology Act 2000. Office of the Gene Technology Regulator, Department of Health and Aging, Australian Government, July 2007. Available at http://bch.cbd.int/database/record-v4.shtml?documentid=101222 (access June 2010).

19. SCBD (2012) Guidance on Risk Assessment of Living Modified Organisms. Final Report of the Ad Hoc Technical Expert Group on Risk Assessment and Risk Management under the Cartagena Protocol on Biosafety, UNEP/CBD/BS/AHTEG-RA&RM/4/6. Available at http://bch.cbd.int/protocol/meetings/documents.shtml?eventid=5037 (access August 2012).

20. The European Parliament and the Council of the European Union (2001) Directive 2001/18/EC of the European Parliament and of the Council of 12 March 2001 on the deliberate release into the environment of genetically modified organisms and repealing Council Directive 90/220/EEC. Available at http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=CELEX:32001L0018:EN:HTML (access June 2010).

21. UNEP (1995) International Technical Guidelines for Safety in Biotechnology. United Nations Environment Programme (UNEP). Available at http://bch.cbd.int/database/record-v4.shtml?documentid=42114 (access June 2010).

22. UNEP Division of Technology, Industry and Economics (website) Technical Workbook on Environmental Management Tools for Decision Analysis. Available at http://www.unep.or.jp/ietc/publications/techpublications/techpub-14/1-EnRA3.asp (access June 2010).

23. UNEP/IPCS (1994) Training module No. 3. Section C – Ecological risk assessment. United Nations Environment Programme (UNEP) / International Programme on Chemical Safety (IPCS), pp 177-222. Available at http://www.chem.unep.ch/irptc/Publications/riskasse/C2text.pdf (access July 2010).

24. US Environmental Protection Agency (1998) Guidelines for Ecological Risk Assessment. EPA/630/R-95/002F. Available at http://oaspub.epa.gov/eims/eimscomm.getfile?p_download_id=36512 (access June 2010).

25. VADLO (website) Biomedical and Life Sciences Search Engine. Available at http://www.vadlo.com (access May 2010).

26. WHO (2004) IPCS Risk Assessment Terminology. Part 2: IPCS Glossary of Key Exposure Assessment Terminology. World Health Organization (WHO). Available at http://www.inchem.org/documents/harmproj/harmproj/harmproj1.pdf (access June 2010).

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