Genetically modi®ed organisms (GMO) in bioremediation andlegislation

Jaroslav DrobnõÂ k*

Association BIOTREND, VinicÏna 5, Praha 2, 128 44 Czech Republic

Received 27 July 1998; received in revised form 31 January 1999; accepted 11 February 1999

Abstract

Some authors expect genetically modi®ed organisms (GMO) to bring a breakthrough in bioremediation. Besides biochemistry

and microbial ecology, legislation and biosafety should be considered in this regard. World wide rules request risk assessment tobe performed before any release of GMO to the environment. Recently, the protocol has been negotiated by UNEP to theConvention of Biological Diversity (CBD), which will be binding to all parties of the CBD. Most attention to the harmonisation

of regulation was paid by OECD. Member as well as non-member states of OECD from central and eastern Europe meetregularly within the framework of UNEP every year. Such harmonisation is vital for international trade, scienti®c activities andco-operation. EU legislation based on directives 90/219/EEC and 90/220/EEC with amendments comprising more directives and

decisions is the general frame for the European legislation. European Standards will be issued shortly. Czech Law on GMO isexpected in the near future and will be based on the EU regulations and protocol to CBD. # 1999 Elsevier Science Ltd. Allrights reserved.

Keywords: Bioremediation; Genetically modi®ed bacteria; Biosafety regulation

1. Why GMO?

The broadest spectrum in metabolic activities is

found in microorganisms. During the course ofevolution archebacteria, eubacteria and fungiacquired the capability to decompose any natural

material. However, this is not so simple when man-made substances are concerned. Plastics are the bestexample.

The removal of man-made pollutants represents themost frequent task of bioremediation. A simple one-

step process is rather rare, usually a sequence of bio-logical transformations is needed to eliminate particu-lar substance. Such a sequence may arise by the

development of a microbial community (called a `con-

sortium') comprising populations with particular bio-transformation activities.

It may happen that in the chain of reactions such asA±B±C±etc., the intermediate B could be more harm-ful than the pollutant A. If the rate B±C is slowerthan the transformation rate A±B, the harmful inter-mediate B will accumulate and the process fails tomeet our expectations.

Genetic engineering o�ers one possibility to solvethis problem. A strain can be constructed either withthe combined capability of converting both A to Band B to C, or with an enhanced expression of an en-zymatic system transforming B to C, and then beintroduced in the natural community.

There are two burdens on this route. The nature ofthe ®rst one lies in the ecology: a laboratory-developedstrain must be designed to ®t into the natural commu-nity. The other comes from legislation: the release of

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* Corresponding author.

organisms developed by genetic engineering is sub-jected to world-wide regulation. Let us brie¯y addressthe latter one.

2. Where do the GMO regulations come from?

Regulation of `genetically modi®ed organisms(GMO)', or `living modi®ed organisms (LMO)', orsimply `products of modern biotechnology' is estab-lished at several levels. The most general level is rep-resented by the United Nations EnvironmentalProgramme (UNEP). It was formulated in the UNEPInternational Technical Guidelines (UNEP, 1995) whichare now used to develop the more bonding Protocol tothe Convention of Biological Diversity. The negotiationsof the latter document are rather di�cult. The 5thmeeting of the open-ended ad hoc working group onbiosafety, established by the conference of parties tothe CBD, will continue the work, starting from August17 1998 in Montreal. The di�erent approaches of thedeveloped and developing countries represent the mainproblem. However, if the protocol is agreed and signedby all parties, it will be bonding for all parties to theCBD.

The Organisation for Economic Co-operation andDevelopment (OECD) has been another internationalorganisation very active in this ®eld. It issued severalfundamental documents (OECD, 1986, 1988, 1991,1992a, 1994, 1995a, 1995b) and organised workshopswhich provided valuable, concise information (OECD,1992b, 1995c, 1995d, 1996a, 1996b; Anon., 1994;Hokkanen and Tsaftaris, 1995). Currently, the en-vironment directorate is producing `consensus docu-ments' summarising all available data on the mostoften used GMO. Documents dealing with the generaPseudomonas, Bacillus and Rhizobium are ready. Veryvaluable for basic information is the database,BioTrack, which can be obtained on http://www.oec-d.org/ehs/service.htm.

Regional rules set another level of regulation. InEurope, such rules are given by EU documents(European Union, 1990a, 1990b) which also im-plemented certain OECD regulations (OECD, 1992c).They are binding for the member states. The non-member countries, as a rule, are developing nationalregulations in harmony with the EU. Among the lattercountries, a law regulating the use and release ofGMO exists in The Russian Federation and Hungary.Czech law on GMO had been prepared by the CzechMinistry of Environment and is expected to be sub-mitted to the Parliament at the end of 1999.

The documents of the Commission and Council ofEU are provided by the European Standards(European Standards, in preparation(a)±(e)). They arecurrently being discussed and, in the near future, they

will be issued by the European Committee forStandardization.

3. What are the principles of the regulations?

Basic di�erences in the principles between theEuropean and American continents consist of criteriaof the object of regulation. In Europe, all organismsdeveloped by the recombinant DNA method should beregulated whereas, in the USA and Canada (also inArgentina), the regulation is based on traits of novelorganisms.

Nevertheless, there are common principles. Therather rigid European principle turned out to be a bur-den to science and technology and generated economi-cal problems. Therefore, new amendments modi®ed itto a more relaxed philosophy. Risk assessment(Mahon, 1995) is the key principle which is in fact thebasis for risk management in all systems of regulation.

Hazard identi®cation is the ®rst step in risk assess-ment. It consists of two components: the nature andthe size of the hazard. The nature of the harm maya�ect the health of man, animal and plants. Also, theimpact on non target organisms, like useful insects,microorganisms, wild life, ®sh etc., must be taken intoaccount. Soil fertility, the ecosystem and the country-side as a whole are other parameters to be included.

The size of the hazard depends on the action envi-saged and organism used. It may be local or general,depending on the organism's ability to spread from thesite of its release.

In the next step, we ask for an assessment of theprobability that the hazard will occur. This is the mostdi�cult task and should be based on experience withsimilar situations, on data from the literature, onmodel experiments, or just formulated as a quali®edguess. As a result of this process, the microorganismand the total envisaged activity has to be classi®ed bya corresponding risk category. Originally, the EUdirective gave two categoriesÐsafe and not safe. Aftera recent amendment, four categories are used in con-formity with risk categories used by WHO.

Because of the uncertainty attached to the par-ameters employed in risk assessment, a precaution fac-tor is usually applied which re¯ects the situation whenall parameters would run in the most unfavourableway.

Based on the risk assessment, corresponding riskmanagement should be designed. It includes the planof the release, the system of controls and a plan forhandling accidents. The regime of the release shouldobey the rules of Good Laboratory Practice (GLP)(OECD, 1992c) or Good Industrial Large ScalePractice (GILSP) designed by OECD (OECD, 1992a).Treatment of the site of the release after the activity is

J. DrobnõÂk / International Biodeterioration & Biodegradation 44 (1999) 3±64

®nished and the monitoring of the site of the releaseafterwards is obligatory.

All governments have established a Competent auth-ority which is the institution to whom the intention ofthe release should be referred. The noti®cation mustinclude both the risk assessment and a plan of riskmanagement in addition to a detailed description ofthe organism and the site of the release. Somecountries also ask for the insurance of the user forpossible damages caused to other parties through therelease. Usually, a written consent from the competentauthority is requested. It is therefore advisable to con-sult the competent authority beforehand.

4. Particular risks in bioremediation

Each group of organisms represent a speci®c risk.Many authorities are particularly reluctant to author-ise the release of genetically modi®ed microorganisms(GMMs) (Sayler and Sayre, 1995). In contrast toplants, little is known about the soil microbial commu-nity (Tiedje et al., 1995) and our tools for monitoringGMM in the soil are very limited. In addition, wehave few data on the gene transfer in soil. All theseuncertainties make the risk assessment very poor andthe risk management is mostly reduced to the standardprotection of personnel; ecology is only covered ingeneral statements. The risk of rambling genes is hardto control unless the strain is not prepared in a specialway to prevent the transfer of the introduced gene.

On the other hand, the transgene is usually derivedfrom another soil microorganism, thus no new gene isadded to the soil microbial community. It is also veryprobable that the introduced engineered strain will notsurvive for long in the soil environment; at least notlong after its speci®c substrate is exhausted. If we takeinto account that the transgen carrying the infor-mation for the speci®c metabolic activity is, as a rule,far from pathogenicity factors, we may conclude thatthe risks of strains designed for bioremediation will bevery low.

5. To use or not to use GMO for bioremediation?

It follows from the above principles that the regu-lations set conditions which cause delay and bringabout additional expenses when GMOs are used forbioremediation irrespective of the risk category of themicroorganism used. Regarding the ecological problemmentioned above, careful consideration and model ex-periments have to be performed before the decision onthe use of GMO is made. On the other hand, the riskassessment could be more reliable and the approach ofcompetent authorities more liberal when practical ex-

perience is accumulating and our familiarity with theuse of GMO in bioremediation is increasing. As in anynew technology, the life of pioneers is not simple.

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