gmo regulations in india - (CUSAT) – Plant Biotechnology laboratory

GMO REGULATIONS ININDIACikesh P.CMicrobial Technology LaboratoryDept.of BiotechnologyCUSAT

• Rules for the manufacture, use/import/export andstorage of hazardous microorganisms/ geneticallyengineered organisms or cells, 1989 notified bythe Ministry of Environment & Forests,Government of India under EnvironmentalProtection Act (1986)• It demands constitution of an InstitutionalBiosafety Committee (IBSC) by everyorganization engaged in research and productionactivities related to GMOs

Department of Biotechnology guidelines "Recombinant DNA Safety Guidelines" (1990DBT Guidelines)"Revised Guidelines for Safety inBiotechnology" (1994 DBT Guidelines)1998 "Revised Guidelines for Research inTransgenic Plants and Guidelines for Toxicity andAllergenicity Evaluation of Transgenic Seeds,Plants and Plant Parts" (1998 DBT Guidelines).

• The 1990 and 1994 DBT guidelinesrecommend appropriate practices,equipments and facilities necessary forsafeguards in handling GMOs in agriculture andpharmaceutical sectors• These guidelines cover‣ R&D activities on GMOs‣ Transgenic crops‣ Large-scale production and deliberaterelease of GMOs‣ Plants, animals and products into theenvironment‣ Shipment and importation of GMOs forlaboratory research

• 1998 DBT guidelines cover areas ofrecombinant DNA research on plants The development transgenic plants and theirgrowth in soil for molecular and field evolution The toxicity and allergenicity data forruminants such as goats and cows, fromconsumption of transgenic plantsData on comparative economic benefits of amodified plant

• Government of India established a three-tierregulatory structure at the central level in NewDelhi comprising three committees-The Review Committee on GeneticManipulation (RCGM) under the Ministry ofScience and Technology (MoST);The Genetic Engineering ApprovalCommittee (GEAC) under the Ministry ofEnvironment and Forestry (MoEF);The Monitoring and Evaluation Committee(MEC) under DBT/MoST

• DBT provides the secretariat for RCGM and MEC,and the MoEF for GEAC.• The GoI setted up a de-centralised structureconsisting of Institutional BiosafetyCommittees (IBSCs) andState and DistrictLevel Committees (SBCCs and DLCs)• IBSCs have been established in all institutions(public and private) that deal with GMOs• By 2008, only three states (out of a total of twentyfivestates and several „union territories‟ that makeup the Indian Union) had created SBCCs• DLCs have not been set up anywhere

• RCGM (Review Committee on GeneticManipulation)‣ assess and decide on the applicationssubmitted by institutions and companies forconducting R&D work,‣ greenhouse tests and contained field tests onplots of less than one acre in size (0.4hectare)Institutions and companies wishing to proceedbeyond these stages towards general release andcommercialisation of GM-crops must conductlarge-scale and multi-location field trials that aremandatory under the biosafety regulations

GEAC (Genetic Engineering ApprovalCommittee)• Has the sole responsibility and power toauthorise large-scale and multi-location fieldtrials• It can asses the „output‟ of the trials and on thebasis of that assessment to approve, reject orput on hold the applicant’s request for generalrelease of the GM-crop for commercial planting,imposing conditions (if need be) under which thegeneral release can take place• GEAC may request ICAR to check and validate the´output‟ of the field trials submitted by theapplicant, if necessary by conducting its own fieldtrials.

MEC (Monitoring and Evaluation Committee)• Monitors the small-scale contained field tests(RCGM’s ambit) and the open larger-scalefield trials (GEAC’s sphere), and submits itsreports to RCGM• MEC‟s monitoring work and reports are expected tocover all the main aspects of biosafety The impact of the GM-crop on theenvironment (ecology and biodiversity)The agronomy (crop production science andfarm-level economy)The health of humans and livestock and thelivelihoods of the farming community

IBSC constitution• (i) Head of the Institution or nominee• (ii) 3 or more scientists engaged in rDNA work ormolecular biology with an outside expert in therelevant discipline.• (iii) A member with medical qualifications -Biosafety Officer (in case of work with pathogenicagents/large scale use).• (iv) A nominee of DBT.

• Every institution setting up an IBSC first needsto identify the members-• The request has to be submitted to DBT alongwith the list and background of the proposedmembers• DBT then identifies an appropriate nominee andcommunicates the same to IBSC chairman.

• If research is conducted on organisms that requirespecial containment conditions (Biosafety Level 3or 4) or if large-scale microbial research isconducted, a Biological Safety Officer must beappointed.• His responsibilities include periodic inspection offacilities and protocols and development ofemergency plan, as higher containment levelsrequire more scrutiny.• This person is also a member of the IBSC

IBSC MEETINGS• Each IBSC has to meet at least twice a year toreview the status of rDNA projects in the institution• The IBSC members are expected to look into thefollowing during the meetings:‣ Action taken on the decisions of earlier IBSCmeetings.‣ Characterization of work and approval as per riskcategory.‣ Evaluation of projects and direction tosubmission for appropriate agencies for approvals

‣ Inspection of containment facilities andgreenhouses etc.‣ Review the medical reports of employees‣ Maintaining procedures and other approvalrequirements.

SUBMISSION OF REPORTS- IBSC• IBSC has to furnish half yearly reports on theongoing projects in the organization to RCGM(Review Committee on Genetic Manipulation)• The report consists of the observance of thesafety guidelines including accidents, risks anddeviations, if any.• There is a prescribed format for informationon projects to IBSC/RCGM and half yearlyreports to be submitted to RCGM

FUNCTIONS- IBSC• The role of IBSCs assumes major importance inthe regulatory framework since it is a StatutoryCommittee that operates from the premises ofthe institution• It is in a position to conduct onsite evaluation,assessment and monitoring of adherence tothe biosafety guidelines.

• The decisions taken by the next highercommittee i.e., Review Committee on GeneticManipulation (RCGM), which operates fromDBT are based on the applications submittedby the investigators with the approval of IBSC• therefore, it is pertinent that the membersof the IBSCs and DBT nominees to theIBSCs have expertise in evaluation,assessment and monitoring of projectsas per the rDNA guidelines.

• Based on the feedback received in theseconsultations, a handbook was prepared byBCIL in consultation with DBT for membersand DBT nominees of IBSCs covering thecomposition, functions, role in approval and achecklist for evaluating projects, which can beused on a case-to-case basis.• A CD containing important biosafetyregulations and guidelines for GMOs is alsoincluded in the handbook for ready referenceby the IBSC members and nominees of DBT

• The IBSC is the nodal point for interactionwithin the institution for implementation of thebiosafety regulatory framework• Highly defined roles for the committees andtheir members• Inorder to strengthen the functioning of thecommittees and their feedback to DBT, DBT inassociation with Biotech Consortium IndiaLimited (BCIL) organized a series of nationalconsultations at six locations namelyBangalore, Chennai, Hyderabad, New Delhi,Mumbai and Jalna

FUNCTIONS- IBSC• Head of the institutionthe biosafety guidelines are followed inhis institution.regular meetings of IBSC are held toreview recombinant research projects inthe institution.open discussion takes place amongst themembers in the meetings and the views ofexternal members as well DBT nomineerecorded.the facilities at the institution are sufficientto meet the containment levels stipulatedfor rDNA products and processes

• IBSC members• The main functions of IBSC members asdefined in the rDNA Safety Guidelines by DBTare as follows:Review and clearance of project proposalsfalling under restricted category, whichfulfill the requirements under the guidelinesTailoring biosafety programme as per thelevel of risk assessed.Training of personnel on biosafety.Instituting a health-monitoring programmefor laboratory personnel.Adopting emergency plans.

• DBT NomineeEach IBSC has a nominee from DBT who oversees theactivities to ensure that safety aspects are being fullyadhered by the organizationThe committee has been constituted as per thenorms of the guidelinesThe Recombinant DNA Safety Guidelines arestrictly followed in the institution.The IBSC meets regularly, at least twice in ayear to review the ongoing activities andprovides half yearly reports to RCGM/DBT in theprescribed performa.All the activities are within the purview of theguidelines and in the knowledge of RCGM/DBT.The DBT nominee is expected to guide the IBSCon biosafety issues.

• Principal Investigator• Depending upon the risk category, the PI has toinform the IBSC, seek permission of IBSC beforestarting the experiments or seek permission ofthe RCGM through its IBSC• The PI is primarily responsible for ensuringcompliance with biosafety standards• The PI functions as a project manager as well asa researcher, communicating with the IBSC andbearing responsibility for training andsupervising personnel

• Based on the nature of the GMO, the PIdetermines the proper containment levelfor the project and, in accordance with theDBT Guidelines, develops the necessaryexperimental protocols.• to make an initial determination of therequired levels of physical and biologicalcontainment in accordance with the DBTguidelines.

to submit the initial research protocol andany subsequent changes (such as changes inthe source of DNA or host vector system) tothe IBSC for review and approval.to ensure that no work is initiated until theresearch project has been approved by the IBSCand has met all requirements of DBT guidelines.remain in communication with the IBSCthroughout the conduct of the project.To ensure the safe conduct of the rDNAexperiments in his laboratory.

To make available the protocols that describethe potential biohazards and the precautions tobe taken to all laboratory staff.To instruct laboratory staff about the practicesand techniques required to ensure safety, andthe procedures for dealing with accidents.To supervise the performance of the laboratorystaff to ensure that the required safety practicesand techniques are employed.To undertake corrective measures promptly forany work errors and conditions that may resultin the release of recombinant DNA materials.

ROLE OF IBSC IN APPROVAL• The rDNA activities within an organizationcould be broadly categorizedresearchlarge-scaleexperiments/production/field releaseimport and shipment

Research• IBSC has to review all recombinant researchcarried out by an organization• The rDNA Safety Guidelines of DBT stipulatethree categories of research activities i.e.Category I, II and III with increasing level ofcontainment requirements• Category I experiments involving self cloning,using strains and also inter species cloningbelonging to organism in the same exchangergroup etc. and are exempt for the purpose ofintimation and approval.

• Category II experiments falling undercontainment levels II, III and IV, large scale useof recombinants made of self cloning in systemsbelonging to exempt category etc. require priorintimation to IBSC• Category III experiments involving toxin genecloning, cloning of genes for vaccine production,use of infectious animals and plant viruses, selffusion experiments, field testing and release etc.require review and approval of IBSC beforecommencement• Depending upon the category of experiments,IBSC can simply note the information provided byPI, give permission before start of theexperiments or forward it to RCGM for approval.

• The categories of genetic engineeringexperiments on plants have been notifiedspecifically under the “Revised Guidelines forResearch in Transgenic Plant, 1998” by DBT• In this categorization, routine recombinant DNAexperiments fall in Category I and need onlyintimation to the IBSC• Category II include lab and greenhouseexperiments in contained environment wheredefined DNA fragments that are non pathogenicto human and animals are used for genetictransformation of plants

• Category III pertains to high risk experimentswhere the escape of transgenic traits into theopen environment could cause significantalterations in the biosphere, the ecosystem,the plants and animals.• All experiments conducted in greenhouse andopen field conditions not belonging to theabove Category II types, would fall underCategory III risks• Such experiments could be conducted onlyafter clearance from RCGM and notified by theDepartment of Biotechnology

• Different levels of containment have beenprescribed for different categories of rDNAexperiments in the guidelines• IBSC should allow genetic engineering activityon classified organisms only at places wheresuch work should be performed as perguidelines• Provision of suitable safe storage facility ofdonor, vectors, recipients and other materialsinvolved in experimental work should be madeand may be subject to inspection onaccountability.

Large scale trials and production• approval for large-scale trials and productionneeds to be taken from GEAC, as per the Rules1989• IBSC has an extremely important role in termsof verifying the information being forwarded toRCGM and GEAC• Both RCGM and GEAC depend on the review ofthe IBSC on the submissions made.• IBSC has to recommend emergency plan incase of large-scale operations includingmethods and procedures for handling largelosses of cultures and organisms

• The interstate shipment of indigenousetiological agents, diagnostic specimens andbiological products need clearance of IBSC• The import of regulated materials forresearch (e.g. toxin genes, hybridomas, cellcultures, organelle) and specifyingconditions under which the agent or vector isshipped, handled and use are issued by RCGM• large scale imports for industrial use areregulated by GEAC• In case of plants, the import is routedthrough the Director, National Bureau ofPlant Genetic Resources on the basis of theimport permit issued by the DBT, based onrecommendations of the RCGM

Information required for risk assessment of a GMOParticularsMolecular Biology DetailsCharacteristics of donor organismsCharacteristics of host/ recipientorganismsInformation requiredOrigin/source and taxonomicclassificationGenome characteristics and priorhistory for genetic manipulationNature of pathogenicity and virulence,infectivity or toxicity, its host range andstability of these traitsTechniques for identification of donororganismOrigin/source and taxonomicclassificationGenome characteristics and priorhistory for genetic manipulation,cultivation/growth and safe useInformation on reproductive cycle(asexual/sexual), its growth andsurvivalNature of pathogenicity and virulence,infectivity or toxicity, its host range andstability of these traits

Characteristics oftransformed/modified organismGenotypic characters includingcharacterization of site ofmodification of recipient genome,regulation and stability of insertedDNA, frequency of mobilization ofinserted DNA and/ or genetic transfercapabilityPhenotypic characters includingtaxonomic characterization,colonization potential, antibioticresistance, infectivity, production oftoxins, allergens, antinutrients or anyother metabolites and its host rangeExpression and properties of thegene product including copies/number of new gene(s), rate andlevel of expression, activity ofexpressed protein, allergenic/toxichazards of the product

Human HealthConsiderationsToxicity• For human food/animal feed,elaborate determination of compositionand assessment of quality oftransformed plants/fruits/seeds aswell as animals, with appropriatecontrolsAllergenicity• Comparison of the characteristicsof the modified organism to therecipient organism regardingallergenicity• Composition analysis of all majoringredients in GMOsNutritional analysis•Changes in the level of key nutrients,natural toxicants or anti nutrients,bioavailability of nutrients.

Environmental ConsiderationsApplication of the modified organismin the environment-Geographical location site-Containment and decontamination-Introduction protocols includingquantity and frequency of application-Methods for monitoring applications- Other including transportarrangements, post-releasemonitoring of the site, impact of GMOon weed formationSurvival, multiplication anddissemination of the GMO in the largescale facilities/field/environment-Description of detection,identification and monitoringtechniques-Specificity, sensitivity and reliabilityof detection techniques-Study of gene flow in plants-Implications of out crossing/crossfertilization

Interactions of modified organismswith biological systems (target andnon-target populations)-Known and predicted habitats of theGMO-Identification and description oftarget organisms and ecosystems-Identification and description of nontargetorganism(s) which might beexposedStability of the GMO in terms ofgenetic traits-Genetic transfer capability-Likelihood of post-release selectionleading to the expression ofunexpected and undesirable traits bythe modified organismRoutes of dissemination-Routes of dissemination, physical orbiological- Known or potential modes ofinteraction, including inhalation,ingestion, surface contact, burrowingand injection

Characteristics of the purifiedproducts from GMOs (e.g.:recombinant therapeutics)- Identity, strength, quality, purity-Evaluation of functional assaysincluding potency of active component,sensitivity, specificity, variability ofassays-Estimates of variability including upperand lower limits for each specificationfor final release of the product- If purified product is held prior tofurther processing, information ondescription of storage conditions andverification of stability

Categorization of microorganisms based on pathogenicityHazard Group 1Hazard Group 2Organisms that are most unlikely to cause human diseaseOrganisms capable of causing human disease and which may bea hazard to laboratory workers, but are unlikely to spread to thecommunity. Laboratory exposure rarely produces infection andeffective prophylaxis or effective treatment is usually availableHazard Group 3Hazard Group 4Organisms that may cause severe human disease and present aserious hazard to laboratory workers. They may present a risk ofspread to the community, but there is usually effective prophylaxisor treatment availableOrganisms that cause severe human disease and are a serioushazard to laboratory workers. They may present a high risk ofspread to the community, and there is usually no effectiveprophylaxis or treatment

BiosafetyLevelPractice and Techniques Safety Facilities1. Standard microbiological practices Non primary containment providedby adherence to standard laboratorypracticesBasic2. Level 1 practices plus laboratorycoats; decontamination of allinfectious wastes limited access;protective gloves and biohazardwarning signs as indicated3. Level 2 practice plus speciallaboratory clothing, controlledaccess4. Level 3 practices plus entrancethrough change room where streetclothing is removed and laboratoryclothing is put on shower on exit,all wastes are decontaminated onPartial containment equipment (i.e.Class I or II Biological SafetyCabinets) used to conductmechanical and manipulativeprocedures that have aerosolpotential that may increase the riskof exposure to personnelPartial containment equipment usedfor all manipulations of infectiousmaterialMaximum containment equipment(i.e. class III biological safetycabinet or partial containmentequipment in combination with fullbody air supplied, positive pressureBasic.ContainmentMaximumcontainment