IS 15889: GUIDELINE FOR THE CONDUCT OF FOOD SAFETY ASSESSMENT OF FOODS PRODUCED USING RECOMBINANT-DNA MICRO-ORGANISMS

2010·0.86 MB·English

by  Bureau of Indian Standards

इंटरनेट मानक
Disclosure to Promote the Right To Information
Whereas the Parliament of India has set out to provide a practical regime of right to
information for citizens to secure access to information under the control of public authorities,
in order to promote transparency and accountability in the working of every public authority,
and whereas the attached publication of the Bureau of Indian Standards is of particular interest
to the public, particularly disadvantaged communities and those engaged in the pursuit of
education and knowledge, the attached public safety standard is made available to promote the
timely dissemination of this information in an accurate manner to the public.
“जान1 का अ+धकार, जी1 का अ+धकार” “प0रा1 को छोड न’ 5 तरफ”
Mazdoor Kisan Shakti Sangathan Jawaharlal Nehru
“The Right to Information, The Right to Live” “Step Out From the Old to the New”
IS 15889 (2010): GUIDELINE FOR THE CONDUCT OF FOOD SAFETY
ASSESSMENT OF FOODS PRODUCED USING RECOMBINANT-DNA
MICRO-ORGANISMS [FAD 23: Biotechnology for Food and
Agriculture]
“!ान $ एक न’ भारत का +नम-ण”
Satyanarayan Gangaram Pitroda
““IInnvveenntt aa NNeeww IInnddiiaa UUssiinngg KKnnoowwlleeddggee””
“!ान एक ऐसा खजाना > जो कभी च0राया नहB जा सकता हहहहै””ै”
Bhartṛhari—Nītiśatakam
“Knowledge is such a treasure which cannot be stolen”

IS 15889 : 2011
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Indian Standard
GUIDELINE FOR THE CONDUCT OF FOOD SAFETY
ASSESSMENT OF FOODS PRODUCED USING
RECOMBINANT-DNA MICRO-ORGANISMS
ICS 67.020
© BIS 2011
B U R E A U O F I N D I A N S T A N D A R D S
MANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG
NEW DELHI 110002
May 2011 Price Group 6
Biotechnology for Food and Agriculture Sectional Committee, FAD 23
FOREWORD
This Indian Standard was adopted by the Bureau of Indian Standards, after the draft finalized by the
Biotechnology for Food and Agriculture Sectional Committee had been approved by the Food and Agriculture
Division Council.
A variety of micro-organisms used in food production have a long history of safe use that predates scientific
assessment. Few micro-organisms have been assessed scientifically in a manner that would fully characterize
all potential risks associated with the food they are used to produce, including, in some instances, the
consumption of viable micro-organisms. Furthermore, the principles of risk analysis, particularly those for
risk assessment, are primarily intended to apply to discrete chemical entities such as food additives and
pesticide residues, or specific chemical or microbial contaminants that have identifiable hazards and risks;
they were not originally intended to apply to intentional uses of micro-organisms in food processing or in the
foods transformed by microbial fermentations. The safety assessments that have been conducted have
focused primarily on the absence of properties associated with pathogenicity in these micro-organisms and
the absence of reports of adverse events attributed to ingestion of these micro-organisms, rather than evaluating
the results of prescribed studies. Further, many foods contain substances that would be considered harmful
if subjected to conventional approaches to safety testing. Thus, a more focused approach is required where
the safety of a whole food is being considered.
This standard has been aligned with the Guideline for the Conduct of Food Safety Assessment of Foods
Produced Using Recombinant-DNA Micro-organisms (CAC/GL 46-2003) and is identical to it.
This approach is based on the principle that the safety of foods produced using recombinant-DNA micro-
organisms, is assessed relative to the conventional counterpart having a history of safe use, not only for the
food produced using a recombinant-DNA micro-organism, but also for the micro-organism itself. This approach
takes both intended and unintended effects into account. Rather than trying to identify every hazard associated
with a particular food, the intention is to identify new or altered hazards relative to the conventional counterpart.
This standard supports the IS 15887 : 2010 ‘Principles for the risk analysis of foods derived from modern
biotechnology’. It addresses safety and nutritional aspects of foods produced through the actions of
recombinant-DNA micro-organisms. The recombinant-DNA micro-organisms that are used to produce these
foods are typically derived using the techniques of modern biotechnology from strains that have a history of
safe, purposeful use in food production. However, in instances where the recipient strains do not have a
history of safe use their safety will have to be established. Such food and food ingredients may contain viable
or non-viable recombinant-DNA micro-organisms or may be produced by fermentation using recombinant-
DNA micro-organisms from which the recombinant-DNA micro-organisms may have been removed.
If a new or altered hazard, nutritional or other food safety concern is identified by the safety assessment, the
risk associated with it would first be assessed to determine its relevance to human health. Following the
safety assessment and if necessary further risk assessment, the food or the component of food, such micro-
organisms used in production would be subjected to risk management considerations in accordance with
IS15887 before it is considered for commercial distribution.
Clauses 3.1.5 and 3.1.6 refer to the concept of ‘substantial equivalence’ as described in the report of the
2000 joint FAO /WHO expert consultations (Document WHO/SDE/PHE/FOS/00.6, WHO, Geneva, 2000).
Report of Joint FAO/WHO Expert Consultation on Allergenicity of Foods Derived from Biotechnology (2001),
which includes reference to several decision trees, was used in developing the Annex to these guidelines.
Clause A-3.3 of this standard may also require reference to Section 6.4 ‘Pepsin Resistance’ of Report of Joint
FAO/WHO Expert Consultation, 2001.
While this Guideline is designed for foods produced using recombinant-DNA micro-organisms or their
components, the approach described could, in g eneral, be applied to foods derived from micro-organisms
that have been altered by other techniques.
IS 15889 : 2011
Indian Standard
GUIDELINE FOR THE CONDUCT OF FOOD SAFETY
ASSESSMENT OF FOODS PRODUCED USING
RECOMBINANT-DNA MICRO-ORGANISMS
1 SCOPE or rendered non-viable during processing; or food
produced using the traditional food production
1.1 The standard describes approaches recommended
micro-organisms for which there is experience of
for making safety assessments of foods produced
establishing safety based on common use in food
using recombinant-DNA micro-organisms, using
production.
comparison to a conventional counterpart, and
identifies the data and information that are generally NOTE — It is recognized that for the foreseeable future,
applicable to making such assessments. micro-organisms derived from modern biotechnology will
not be used as conventional counterparts.
1.2 This standard does not address,
3 INTRODUCTION TO FOOD SAFETY
a) safety of micro-organisms used in ASSESSMENT
agriculture (for plant protection,
3.1 General
biofertilizers, in animal feed or food derived
from animals fed the feed, etc); 3.1.1 Most foods produced as a result of the
purposeful growth of micro-organisms have their
b) risks related to environmental releases of
origins in antiquity, and have been deemed safe long
recombinant-DNA micro-organisms used in
before the emergence of scientific methods for
food production;
assessing safety. Micro-organisms possess
c) safety of substances produced by micro-
properties, such as fast growth rates, that enable
organisms that are used as additives or
genetic modifications, whether employing
processing aids, including enzymes for use
conventional techniques or modern biotechnology,
in food production;
to be implemented in short time frames. Micro-
d) specific purported health benefits or organisms used in food production derived using
probiotic effects that may be attributed to conventional genetic techniques have not
the use of micro-organisms in food; or customarily been systematically subjected to
e) issues relating to the safety of food extensive chemical, toxicological, epidemiological, or
production workers handling recombinant- medical evaluations prior to marketing. Instead
DNA micro-organisms. microbiologists, mycologists, and food
technologists have evaluated new strains of bacteria,
1.3 The criterion for establishing the safety of micro- yeasts and filamentous fungi for phenotypic
organisms used in the production of foods where characteristics that are useful in relation to food
there is no history of safe use is beyond the scope production.
of this standard.
3.1.2 Safety assessments of recombinant-DNA
2 DEFINITIONS micro-organisms should document the use of related
micro-organisms in foods, the absence of properties
For the purpose of this standard, the following
known to be characteristic of pathogens in the
definitions shall apply.
recombinant-DNA micro-organisms or the recipient
2.1 Recombinant-DNA Micro-organism — Bacteria, strains used for constructing the recombinant-DNA
yeasts or filamentous fungi in which the genetic micro-organisms, and known adverse events
material has been changed through in vitro nucleic involving the recipient or related organisms. In
acid techniques including recombinant addition, when a recombinant DNA micro-organism
deoxyribonucleic acid (DNA) and direct injection of directly affects or remains in the food, any effects
nucleic acid into cells or organelles. on the safety of the food should be examined.
2.2 Conventional Counterpart — A micro-organism/ 3.1.3 The use of animal models for assessing
strain with a known history of safe use in producing toxicological effects is a major element in the risk
and/or processing the food and related to the assessment of many compounds, such as pesticides.
recombinant-DNA strain. The micro-organism may In most cases, however, the substance to be tested

be viable in the food or may be removed in processing is well characterized, of known purity, of no particular
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IS 15889 : 2011
nutritional value, and human exposure to it is 3.1.5 Due to the difficulties of applying traditional
generally low. It is therefore relatively straightforward toxicological testing and risk assessment procedures
to feed such compounds to animals at a range of to whole foods, a more focused approach is required
doses some several orders of magnitude greater than for the safety assessment of foods produced using
the expected human exposure levels, in order to recombinant-DNA micro-organisms. This has been
identify any potential adverse health effects of addressed by the development of a multidisciplinary
importance to humans. In this way, it is possible, in approach for assessing safety that takes into account
most cases, to estimate levels of exposure at which the intended effect, the nature of the modification
adverse effects are not observed and to set safe and detectable unintended changes that may occur
intake levels by the application of appropriate safety in the micro-organism or in its action on the food,
factors. using the concept of substantial equivalence.
3.1.4 Animal studies cannot readily be applied to 3.1.6 While the focus of a safety assessment will be
testing the risks associated with whole foods, which on the recombinant-DNA micro-organism, additional
are complex mixtures of compounds, and often information on its interaction with the food matrix
characterized by a wide variation in composition and should be taken into consideration when applying
nutritional value. Due to their bulk and effect on the concept of substantial equivalence, which is a
satiety, they can usually only be fed to animals at key step in the safety assessment process. However,
low multiples of the amounts that might be present the concept of substantial equivalence is not a safety
in the human diet. In addition, a key factor to consider assessment in itself. Rather it represents the starting
in conducting animal studies on foods is the point that is used to structure the safety assessment
nutritional value and balance of the diets used, in of both a recombinant-DNA micro-organism relative
order to avoid the induction of adverse effects that to its conventional counterpart and the food
are not related directly to the material itself. Detecting produced using recombinant-DNA micro-organism
any potential adverse effects and relating these relative to its conventional counterpart. This concept
conclusively to an individual characteristic of the is used to identify for evaluation similarities and
food can therefore be extremely difficult. If the differences between a recombinant-DNA micro-
characterization of the food indicates that the organisms used in food processing as well as the
available data are insufficient for a thorough safety food produced using the recombinant-DNA micro-
assessment, properly designed animal studies could organisms and their respective conventional. It aids
be requested on the whole food. Another in the identification of potential safety and nutritional
consideration in deciding the need for animal studies issues and is considered the most appropriate
strategy to date for safety assessment of foods
is whether it is appropriate to subject experimental
produced using recombinant-DNA micro-organisms.
animals to such a study if it is unlikely to give rise to
The safety assessment carried out in this way does
meaningful information.
not imply absolute safety of the new product; rather,
Animal studies typically employed in toxicological it focuses on assessing the safety of any identified
evaluations also cannot be readily applied to testing differences so that the safety of the recombinant-
potential risks associated with ingestion of micro- DNA micro-organism and the food produced using
organisms used for food production. Micro- recombinant-DNA micro-organism can be considered
organisms are living entities, containing complex relative to their respective conventional
structures composed of many biochemicals, and counterparts.
therefore are not comparable to pure compounds. In
3.2 Unintended Effects
some processed foods, they can survive processing
and ingestion and can compete and, in some cases, 3.2.1 In achieving the objective of conferring a
be retained in the intestinal environment for specific target trait (intended effect) to a micro-
significant periods of time. Appropriate animal organism by the addition, substitution, removal, or
studies should be used to evaluate the safety of rearrangement of defined DNA sequences, including
recombinant-DNA micro-organisms where the donor, those used for the purpose of DNA transfer or
or the gene or gene product do not have a history of maintenance in the recipient organism, additional
safe use in food, taking into account available traits could, in some cases, be acquired or existing
information regarding the donor and the traits could be lost or modified. The potential for
characterization of the modified genetic material and occurrence of unintended effects is not restricted to
the gene product. Further, appropriately designed the use of in vitro nucleic acid techniques. Rather, it
studies in animals may be used to assess the is an inherent and general phenomenon that can also
nutritional value of the food or the bioavailability of occur in the development of strains using traditional
the newly expressed substance in the food. genetic techniques and procedures, or from exposure

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of micro-organisms to intentional or unintended relevance and potential impact on food safety. A
selective pressures. Unintended effects may be variety of data and information is necessary to assess
deleterious, beneficial, or neutral with respect to unintended effects, because no individual test can
competition with other micro-organisms, ecological detect all possible unintended effects or identify, with
fitness of the micro-organism, the micro-organism’s certainty, those relevant to human health. These data
effects on humans after ingestion, or the safety of and information, when considered in total, should
foods produced using the micro-organism. provide assurance that the food is unlikely to have
Unintended effects in recombinant-DNA micro- an adverse effect on human health. The assessment
organisms may also arise through intentional of unintended effects takes into account the
modification of DNA sequences or they may arise biochemical, and physiological characteristics of the
through recombination or other natural events in the micro-organism that are typically selected for
recombinant-DNA micro-organism. Safety improving strains for commercial food or beverage
assessment should include data and information to uses. These determinations provide a first screen
reduce the possibility that a food derived from a for micro-organisms that exhibit unintended traits.
recombinant-DNA micro-organism would have an Recombinant-DNA micro-organisms that pass this
unexpected, adverse effect on human health.
screen are subjected to safety assessment as
described in 4.
3.2.2 Unintended effects can result from the
insertion of DNA sequences new to a micro-organism
3.3 Framework of Food Safety Assessment
into the microbial genome; they may be compared
with those observed following the activity of 3.3.1 The safety assessment of a food produced
naturally occurring transposable genetic elements. using a recombinant-DNA micro-organism is based
Insertion of DNA may lead to changes in expression on determining the safety of using the micro-
of genes in the genome of the recipient. The insertion organism, which follows a stepwise process of
of DNA from heterologous sources into a gene may addressing relevant factors that include,
also result in the synthesis of a chimeric protein,
a) description of the recombinant-DNA micro-
also referred to as a fusion protein. In addition genetic
organism;
instability and its consequences need to be
considered. b) description of the recipient micro-organism
and its use in food production;
3.2.3 Unintended effects may also result in the
formation of new or changed patterns of metabolites.
c) description of the donor organism(s);
For example, the expression of enzymes at high levels
or the expression of an enzyme new to the organism d) description of the genetic modification(s)
may give rise to secondary biochemical effects, including vector and construct;
changes in the regulation of metabolic pathways, or
e) characterization of the genetic
altered levels of metabolites.
modification(s); and
3.2.4 Unintended effects due to genetic modification
f) safety assessment:
may be subdivided into two groups: those that could
be predicted and those that are “unexpected”. Many 1) expressed substances: assessment of
unintended effects are largely predictable based on potential toxicity and other traits related
knowledge of the added trait, its metabolic to pathogenicity;
consequences or of the site of insertion. Due to the
expanding knowledge of microbial genomes and 2) compositional analyses of key
physiology, and the increased specificity in function components;
of genetic materials introduced through recombinant-
3) evaluation of metabolites;
DNA techniques compared with other forms of
genetic manipulation, it may become easier to predict 4) effects of food processing;
unintended effects of a particular modification.
5) assessment of immunological effects;
Molecular biological and biochemical techniques can
also be used to analyse changes that occur at the
6) assessment of viability and residence
level of transcription and translation that could lead
of micro-organisms in the human
to unintended effects.
gastrointestinal tract;
3.2.5 The safety assessment of foods produced
7) antibiotic resistance and gene transfer;
using recombinant-DNA micro-organisms involves
and
methods to identify and detect such unintended
effects and procedures to evaluate their biologi cal 8) nutritional modification.
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3.3.2 In certain cases, the characteristics of the contained in food should be conserved as stock
micro-organisms and/or the foods produced/ cultures with appropriate identification using
processed using these micro-organisms may molecular methods, and preferably, in established
necessitate generation of additional data and culture collections. This may facilitate the review of
information to address issues that are unique to the the original safety assessment. Such stock cultures
micro-organisms and/or food products under review. should be made available to regulatory authorities
Experiments intended to develop data for safety upon request.
assessments should be designed and conducted in
4.2 Description of the Recipient Micro-organism
accordance with sound scientific concepts and
and Its Use in Food Production
principles, as well as, where appropriate, good
laboratory practice. Primary data should be made 4.2.1 A comprehensive description of the recipient
available to regulatory authorities upon request. micro-organism or micro-organism subjected to the
Data should be obtained using sound scientific modification should be provided. Recipient micro-
methods and analysed using appropriate statistical organisms should have a history of safe use in food
techniques. The sensitivity of all analytical methods production or safe consumption in foods. Organisms
should be documented. that produce toxins, antibiotics or other substances
that should not be present in food, or that bear
3.3.3 The goal of each safety assessment is to
genetic elements that could lead to genetic
provide assurance, in the light of the best available
instability, antibiotic resistance or that are likely to
scientific knowledge, that the food will not cause
contain genes conferring functions associated with
harm when prepared or consumed according to its
pathogenicity (that is also known as pathogenicity
intended use, nor should the organism itself cause
islands or virulence factors) should not be
harm when viable organisms remain in the food.
considered for use as recipients. The necessary data
Safety assessments should address the health
and information should include, but need not be
aspects for the whole population, including immuno-
restricted to:
compromised individuals, infants, and the elderly.
The expected endpoint of such an assessment will a) identity: scientific name, common name or
be a conclusion regarding whether the new food and/ other name(s) used to reference the micro-
or micro-organisms are as safe as the conventional organism, strain designation, information
counterparts taking into account dietary impact of about the strain and its source, or accession
any changes in nutritional content or value. Where numbers or other information from a
the micro-organism is likely to be viable upon recognized culture repository from which the
ingestion, its safety should be compared to a organism or its antecedents may be
conventional counterpart taking into account obtained, if applicable, information
residence of the recombinant-DNA micro-organism supporting its taxonomical assignment;
in the gastrointestinal tract, and where appropriate,
b) history of use and cultivation, known
interactions between it and the gastrointestinal flora
information about strain development
of mammals (especially humans) and impacts of the
(including isolation of mutations or
recombinant-DNA micro-organism on the immune
antecedent strains used in strain
system. In essence, the outcome of the safety
construction); in particular, identifying traits
assessment process is to define the product under
that may adversely impact human health;
consideration in such a way as to enable risk
c) information on the recipient micro-
managers to determine whether any measures are
organism’s genotype and phenotype
needed to protect the health of consumers and if so
relevant to its safety, including any known
to make well-informed and appropriate decisions in
toxins, antibiotics, antibiotic resistance
this regard.
factors or other factors related to
4 GENERAL CONSIDERATIONS pathogenicity, or immunological impact, and
information about the genetic stability of the
4.1 Description of the Recombinant-DNA Micro-
micro-organism;
organism
d) history of safe use in food production or
A description of the bacterial, yeast, or fungal strain safe consumption in food; and
and the food being presented for safety assessment
e) information on the relevant production
should be provided. This description should be
parameters used to culture the recipient
sufficient to aid in understanding the nature of the
micro-organism.
organism or food produced using the organism being
submitted for safety assessment. Recombinant-DNA 4.2.2 Relevant phenotypic and genotypic
micro-organisms used in food production or information should be provided not only for the
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recipient micro-organism, but also for related species identification of all genetic material potentially
and for any extra-chromosomal genetic elements that delivered to or modified in the recipient micro-
contribute to the functions of the recipient strain, organism and to provide the necessary information
particularly if the related species are used in foods for the analysis of the data supporting the
or involved in pathogenic effects in humans or other characterization of the DNA added to, inserted into,
animals. Information on the genetic stability of the modified in, or deleted from the microbial genome.
recipient micro-organism should be considered
4.4.2 The description of the strain construction
including, as appropriate, the presence of mobile
process should include,
DNA elements, that is, insertion sequences,
transposons, plasmids, and prophages. a) information on the specific method(s) used
for genetic modification;
4.2.3 The history of use may include information
on how the recipient micro-organism is typically b) information on the DNA used to modify the
grown, transported and stored, quality assurance micro-organism, including the source (for
measures typically employed, including those to example plant, microbial, viral, synthetic),
verify strain identity and production specifications identity and expected function in the
for micro-organisms and foods, and whether these recombinant-DNA micro-organism, and
organisms remain viable in the processed food or copy number for plasmids; and
are removed or rendered non-viable as a c) intermediate recipient organisms including
consequence of processing. the organisms (for example other bacteria or
fungi) used to produce or process DNA prior
4.3 Description of the Donor Organism(s)
to introduction into the final recipient
Information should be provided on the donor organism.
organism(s) and any intermediate organisms, when
4.4.3 Information should be provided on the DNA
applicable, and, when relevant, related organisms. It
added, inserted, deleted, or modified, including,
is particularly important to determine if the donor or
intermediate organism(s) or other closely related a) characterization of all genetic components
species naturally exhibit characteristics of including marker genes, vector genes,
pathogenicity or toxin production, or have other regulatory and other elements affecting the
traits that affect human health. The description of function of the DNA;
the donor or intermediate organism(s) should include:
b) size and identity;
a) identity: scientific name, common name or c) location and orientation of the sequence in
other name(s) used to reference the the final vector/construct; and
organism, strain designation, information d) function.
about the strain and its source, or accession
numbers or other information from a 4.5 Characterization of the Genetic Modification(s)
recognized culture repository from which the
4.5.1 In order to provide clear understanding of the
organism or its antecedents may be
impact of the genetic modification on the composition
obtained, if applicable, and information
and safety of foods produced using recombinant-
supporting its taxonomic assignment;
DNA micro-organisms, a comprehensive molecular
b) information about the organism or related and biochemical characterization of the genetic
organisms that concerns food safety; modification should be carried out. To facilitate the
c) information on the organism’s genotype and safety assessment, the DNA to be inserted should
phenotype relevant to its safety including any be preferably limited to the sequences necessary to
known toxins, antibiotics, antibiotic resistance perform the intended functions.
factors or other factors related to pathogenicity,
4.5.2 Information should be provided on the DNA
or immunological impact; and
modifications in the recombinant DNA micro-
d) information on the past and present use, if
organism; this should include,
any, in the food supply and exposure
route(s) other than intended food use (for a) characterization and description of the
example, possible presence as added, inserted, deleted, or otherwise
contaminants). modified genetic materials, including
plasmids or other carrier DNA used to
4.4 Description of the Genetic Modification(s)
transfer desired genetic sequences. This
Including Vector and Construct
should include an analysis of the potential
4.4.1 Sufficient information should be provided on for mobilization of any plasmids or other
the genetic modification(s) to allow for the genetic elements used, the locations of the
5