Ministry of Science and Technology-MCT
National Technical Biosafety Commission-CTNBio

TECHNICAL OPINION No.1255/2008.

Process No.:01200.002109/2000-04
Petitioner: Syngenta Seeds Ltda.
CNPJ: 049.156.326/0001-00
Address: Av. das Nações Unidas, 1801 – 4º andar - São Paulo
– SP – CEP 04795-900
Subject: Commercial Release of Genetically Modified Corn
Previous Extract: Communication No.115/2000 published on
Jul/31/2000.
Meeting: 106th Ordinary Meeting of CTNBio that took place
on September 20th, 2007.
Decision: GRANTED

After appreciation of request for Technical Opinion for
commercial liberation of genetically modified corn
resistant to insects of the Lepidoptera order (Bt11 corn,
Event Bt11), as well as all the progenies coming from the
transformation event Bt11, and its derivatives of lineages
crossings, and non—transgenic populations of corn with
lineages bearing event Bt11, CTNBio decided to GRANT it, on
the terms of this conclusive technical opinion.
Syngenta Seeds Ltda. requested from CTNBio a Technical
Opinion for the free registration, use, essays, tests,
seeding, transportation, storage, commercialization,
consume, importation, liberation and discard of corn (Zea
mays, L.) resistant to insects of the Lepidoptera order –
Corn Bt11. This corn was genetically modified through the
insertion of plasmid pZO1502 containing a fusion of gene
cry1A(Btk) with gene pat. The event of transgenic Bt11 corn
was obtained through the direct transfer of DNA in
protoplasts of lineage H8540 of corn, deriving from embryo
cells in culture in suspension treated with enzymes for
degradation of cellular wall. It contains the synthetic
gene Btk, that comes from Bacillus thuringiensis var.
kurstaki, that codifies á-endotoxin Cry 1Ab, that enables
the translation of á-endotoxin lethal to insects that
ingest these cells, particularly those of Lepidopterus
order, and gene pat, derived from Streptomycin
viridochromogenes cepa Tu494, and the codifier of the
phosphinotricin enzyme N-acetyltransferase (PAT). For
Cry1A(b), the highest expression levels were observed on
leaves, with 27 to 33 μg/g of fresh tissue. Levels 5 to 10
times lower were observed in straw tissues, stem and
grains. For PAT, the amount described on leaves is of the
order of 44 ng/g of fresh tissue. Half of this value was
found on panicles, and 10 times less in style-stigmas. No
allergenic or toxic effects were pointed out coming from
genetically modified plants and grains. Genetically
modified proteins are degraded by digestion of food, by
gastric fluids and by bacteria present on human being and
animals’ gastrointestinal treat. Due to plants bigger
production to the attack of insects, and, particularly, of
Bt11 corn spikes, there are less toxins of fungus origin in
grains, reducing the possibility of intoxications of human
beings and animals. Proteins Cry and PAT do not become
volatile, nor are absorbed by the epidermis, and,
therefore, it would not be justifiable to evaluate the
toxicity of such proteins through inhaling or via dermis.
No unintentional meaningful biological change occurred on
the composition, or on the nutritious value of the grain,
and of the Bt11 corn sawdust, as a consequence of Cry1A(b),
and pat transgene expression, suggesting, then, that Bt11
corn is substantially equivalent in nutritious composition
to the respective isogenic hybrid not genetically modified
and commercial hybrids of corn. The dispersion of corn
seeds is easily controlled, once corn domestication
eliminated the ancestral mechanisms of seeds dispersion,
and pollen movement is the only effective escape mean of
corn plants genes. The horizontal gene flow between Bt and
other species, even those that are very related, have
almost no probability of occurrence, for sylvan species
related to corn do not naturally occur in Brazil. The
coexistence between conventional corns cultivations
(improved or creoles), and transgenic cultivation is
possible from the agronomic point of view, and for that,
one should observe the disposition on Normative Resolution
No. 4 of CTNBio. Once B. thuringiensis is a soil
microorganisms, the exposition of live organisms, and of
the environment to this bacteria, or to any element
extracted from it, is an event that abundantly occurs in
nature, not resulting in meaningful risk for the soil micro
biota. However, even if genic flow occurs between Bt11 corn
plants and the creoles varieties, differences of the gene
flow in relation to any other existing allele in plants are
expected. In sum, the gene or allele will only stay in the
population if the gene flow is continuous, with relatively
high frequency, and if there is any adaptation advantage.
In the Brazilian environment, where sexually compatible
native species do not occur, or are known, the risk that
Bt11 corn execute or promote the invasion of uncultivated,
and cultivated areas does not exist. The ingestion of
endotoxin Cry1A(b) by worms of Spodoptera frugiperda,
Helicoverpa zea and Ditrea saccharalis with alkaline
digestive environment will promote its death through the
interaction of the protein with the receptors of cellular
surface of intestinal cells of these insects, promoting the
opening of the pores, and the invasion of the
microorganisms in the intestinal treat. Thus, insects’
death derives from the osmotic unbalance promoted by the
toxin, and by septicemia deriving from the invasion of
microorganisms into the intestinal flora. Meaningful
differences were not observed between the populations of
ladybugs, carabidae, cincidelidae and spiders, neither of
parasitoid of H. zea, Trichogramma sp., when Bt11 plants
were compared to their genetically unmodified isogenic
lineage. Bt11 hybrids were efficient for the control of the
evaluated plague-lepidopteron, and superior for the profit
agronomic parameters of grains and of bitter grains. For
the other evaluated agronomic parameters (plants height,
insertion height of spikes, date of male and female
flowering, note for diseases, percentage of erect plants,
kind of grain, grain color), Bt11 hybrids presented
performance statistically equals to the respective isogenic
not GM hybrids, confirming the equivalence of agronomic
performance between B11 hybrids, and the non GM isogenic in
conditions of the culture cultivation in Brazil. In Brazil,
nowadays, there is an indiscriminate use of insecticides,
and even a mixture of chemical products, to try to control
insects, especially S. frugiperda. The use of Bt technology
in Brazil may contribute for the reduction of the use of
insecticides, and, consequently, reduce the impacts of the
use of such agro toxics in the environment, in human and
animal’s health, and it may also indirectly help on the
preservation of untargeted organisms’ populations, and
benefic insects, facilitating the integrated handling of
crop plagues. The use of genetically modified plants
resistant to insects present positive repercussions also in
the aspects related to the acquisition and use of chemical
insecticides, to meaningfully reduce the pollution provoked
by industrial rejects, and by the use of water used on
pulverizations, besides avoiding man, food, rivers and
springs contamination deriving from the use, transportation
and storage of insecticides. Before the foregoing, one can
conclude that the cultivation and consume of Bt11 corn is
not the potential cause of meaningful degradation of the
environment, or of risks to human and animal’s health. For
these reasons, there are no restrictions to the use of this
corn, or its derivatives. The petitioner should conduct
monitoring after the commercial release on the terms of
Normative Resolution No.3 of CTNBio. In accordance with
what is established on art. 1 of law 11.460, of March 21st,
2007, “it is vetoed the research and cultivation of
organisms genetically modified on indigenous lands and
areas of conservation units”. In the ambit of competences
of art. 14 of Law 11.105/05, CTNBio considered that the
request fulfills the norms and the pertinent legislation
that aim at guaranteeing biosafety of the environment, of
agriculture and of human and animal’s health.

CTNBio’s TECHNICAL OPINION

I. GMO Identification
Designation of GMO: Bt11 Corn
Petitioner: Syngenta Seeds Ltda.
Species: Zea mays L.
Inserted Characteristics: Resistance to insects of
Lepidoptera order
Method of characteristic introduction: Direct
transformation of protoplasts
Proposed Use: Silage and grains production for human and
animal consume of GMO, and its derivatives.
II. General Information
Corn Zea mays L. is a species from the Gramineae family,
Maydae tribe, Panicoideae family. Corn is a separate
species within Zea sub-gender, with chromosome number 2n =
20, 21, 22, 24 (26). The sylvan species closer to corn is
teosinte, found in Mexico, and in some places in Central
America, where it can be crossed with corn cultivated in
production fields. The corn produced can also be crossed
with the most distant genre Tripsacum. This crossing,
however, occurs with great difficulty and results on
sterile-male progeny.
Corn history is over eight thousand years old in the
Americas, being cultivated since the pre-Colombian period.
It is one of the superior plants best scientifically
characterized, being, nowadays, the cultivated species that
reached the highest degree of domestication, and only
survives in nature when it is cultivated by men(4). Today,
there are around 300 races of corn, and within each race,
thousands of crops.
Corn is one of the most important sources of food in the
world, and is raw material for the production of a wide
range of food products, rations and industrial products.
Brazil is the third biggest corn producer in the world with
a production of approximately 35 million tons in 2005,
behind only of the United States of America (282 million
tons), and China (139 million tons)(29). In Brazil, corn is
basically planted in two crops (summer plantation, and
small crop), and it is cultivated practically all over the
national territory, being 92% of the production
concentrated in the South (47% of production), Southeast
(21% of production) and Center-West (24% of production)
(19). In the productive chain of swine, and poultry,
approximately 70 to 80% of the corn produced in Brazil is
consumed.
It is known that the occurrence of insects in the tropics
is bigger than the one in tempered climate regions, and
that damages caused are more accentuated. Among the most
important corn plagues, one can highlight Spodoptera
frugiperda. Cruz et al.(21) estimated that the loss in
Brazil, due to the infestation by S. frugiperda was around
400 million dollars per year. From 1999, it was observed an
increase on the occurrence of S. frugiperda, and
consequently there was increment on the harms. Other
species of Lepidoptera order are also important plagues for
corn cultivation, such as Helicoverpa zea, and Diatrea
saccharalis). It is estimated that these three species may
cause damages of up to 34% on corn grains production.
The main insects control measure on corn culture has been
the insecticides use. In some areas of the Brazilian
center-West regions, for example, dozens of pulverizations
with insecticides are necessary in only one culture cycle.
Another plague control measure would be the use of
resistant cultivars. The acquisition of cultivars resistant
to insects through classic genetic improvement has not
obtained the hoped success. In the case of S. frugiperda,
many attempts have been made with limited success(77).
Brazil is the third biggest consumer of agricultural
defensives in the world. Nowadays, we have 142 agro toxics
registered for corn, 107 only for worms. There are already
many cases of resistance for the constant and
indiscriminate use of insecticides in corn culture in
Brazil. Besides, one of the factors that affects
agriculturists’ health the most in Brazil is the use of
agricultural defensives responsible for the intoxication of
a million people every year (2).
Bt11 genetically modified corn presents characteristics
that confer resistance, on the same plant, to insects, and
to glufosinate of ammonium herbicide, and resists to the
main plagues of Lepidoptera Order that affect corn culture
in Brazil, such as S. frugiperda, and H.zea. The genes
introduced codify an incomplete form of Bt insecticide
protein, obtained from cepa HD-1 of the soil bacteria
Bacillus thuringiensis var. kurstaki (btk), and an enzyme
(phosphinotricin-N-acetyl transferase, PAT), that confers
tolerance to glufosinate of ammonium herbicide, also
obtained from a soil bacteria, Streptomyces
viridochromogenes. Varieties of corn containing Cry
proteins have been used in many countries in the world, and
there is no information that hybrids of corn containing cry
genes have caused damage to the environment, or to human
and other animals’ health. Corn Bt11 is commercialized in
16 countries (Argentina, Australia, Canada, China, European
Union, Japan, Korea, Mexico, Philippines, Russia, South
Africa, Switzerland, Taiwan, The United Kingdom, The United
States of America, and Uruguay), being commercially
cultivated in the United states (1996), Canada (1996),
Japan (1996), South Africa (2003), Philippines (2005),
Argentina (2001) and Uruguay (2004).
In Brazil many necessary experiments were conducted, and
enough studies were made to convince CTNBio’s members about
the biosafety of the event in study. In the risk analysis,
the molecular characterization should be considered, taking
also in consideration studies carried out regarding the
constitutional, agronomic, and physiologic
characterization, of this event itself. The long experience
with traditional methods of plants improvement, the
experience of over three decades in research, and more than
one decade of commercialization of transgenic varieties in
the world, besides the advancement in the knowledge about
the structure and dynamics of genomes, indicating if a
certain gene, or characteristics is safe, signal that the
process of genetic engineering on its own presents little
potential for arising unexpected consequences that would
not be identified, or eliminated during the process of
genetically modified varieties development (8).
III. Description of GMO and Expressed Proteins
Bt11 corn was genetically modified through the insertion of
plasmid pZ01502 containing the fusion of gene cry1A (Btk)
with gene pat. This corn expresses gene cry1A(b), derived
from the soil bacteria B. thuringiensis subsp. kurstaki,
lineage HD-1.
B. thuringiensis (Bt) is a gram positive bacteria of
Bacillaceae family that produces, at the moment of its
sporulation, crystalline proteic inclusions. These
inclusions contain proteins called á-endotoxins, which
nowadays form a family of 300 members, classified in 49
groups(20). They are produced under the form of prototoxins
that are transformed into toxic peptides in the
insect’s intestines, through the action of intestinal
alkaline pH, and of proteases. The active toxin causes the
destruction of epithelial cells, and the death of the
larvae(47,23). B. thuringiensis may be considered the
biological agent of greatest potential for the control of
forests, agricultural plague-insects, and vectors of
diseases, thanks to the specificity of the á-endotoxins to
insects and target-invertebrate, and its innocuousness to
vertebrates and to the environment, including benefic
insects and natural enemies(43), making this agent a keycomponent
in strategies of integrated handling of
plagues(59).
The event of Bt11 transgenic corn was obtained through the
direct transfer of DNA(nu) in protoplasts of lineage H8540
of corn, derived from embryo cells in culture in suspension
treated with enzymes for degradation of the cell wall, and
it contains DNA sequences inserted into the cell genome,
according to the following description. The synthetic gene
Btk codifies á-endotoxin cry1Ab. The objective of use of
Btk genic cassette is to allow, in vegetable cells, the
transcription of RNA, and the translation of lethal á-
endotoxin to insects that ingest these cells, particularly
those of the Lepidoptera order, such as the ones of
Spodoptera, Helicoverpa and Diatrea genre (61, 17, 34).
Modifications on the original sequence of Btk were carried
out in order to alter some codons of preferential use in
bacteria for the preferential pattern of vegetable codons,
as well as the truncation, that is, the reduction of the
size of the codifying sequence, in order to produce a more
effectively toxic version to targeted-insects. The
synthetic nucleotide sequence, on the truncation version
did not alter the polypeptide sequence of codified protein
on the considered region. The final sequence of gene
Btk(1845 pb) illustrated on the process allows for its
immediate comparison with the original sequences of
cry1A(b) of B. thuringiensis var.kurstaki available at
GenBank with those under the access codes AYB47289, and
AFO59670, among others (65,71,39). Gene Btk is regulated by
two nucleotide sequences upstream, constituted by the
promoter RNA35S of mosaic virus of cauliflower (35S CaMV),
isolated CM1841 with 514 pb, and the intron sequence IV56
of gene of 1S (Adh1S) desidrogenase alcohol of corn, with
412 pb. With these regulating elements the transcription of
gene Btk has its potential increased in vegetable cells. As
terminating sequence, the cassette of expression has a
terminal region of 270 pb of gene of nopalina-syntase (3’-
nos) of T-DNA of Aggrobacterium tumefciens. All the
regulating elements of the transcription have a function
widely described in scientific literature (45, 35, 48). In
the case of a insecticide toxin without known, or described
enzymatic activity, one cannot expect metabolic alterations
deriving from the expression of Btk in vegetable cells. The
measures of general metabolic contents reinforce the idea
that, if any chemical alteration occurs due to genetic
transformation, it is not perceptible through sensible
methods of analysis, such as, for example, spectroscopy of
near infra-red (NIRS).
Another component of Bt11 corn is gene pat, derived from
Streptomycin viridochromogenes cepa Tu494 and codifier of
phosphinotricin N-acetyltransferase (PAT) enzyme. The
original sequence was modified to reduce the content G/C
and alter the beginning of the translation GTG to ATG, in
order to enable, and optimize the syntheses of the original
protein. The final version of pat gene has 558 pb. Again, a
551 pb sequence of the promoter 35S of CaMV (isolated Cabbs),
and the intron sequence IVS2 of 178 pb of gene adhS1 of
corn were used to promote and increase the transcription of
pat gene. Sequence 3’-nos of 220 pb was used as a
terminator element of transgenes. This cassette allows,
then, the syntheses of the recombinant protein PAT, capable
of chemically inactivate herbicides deriving from
phosphinotricin, such as glufosinate of ammonium, making
cells and vegetables that contain it resistant to it. Pat
Enzyme has described and well-known activity (32, 57, 70).
Bt11 corn has framework of plasmid pUC18, including the
origin of replication, and places of recognition of endonucleases
that allow the adaptation of sequences. These
vector DNA fragments have 1520 pb of extension and there is
no evidence that they are expressed on vegetable cells(7).
The final version of the plasmid used on the genetic
transformation of corn was called pZ01502, and has 6,120
pb, including all the cassettes and elements of DNA
described above. This plasmid was destitute from the gene
of bacteria resistance to antibiotics derived from
penicillin, such as ampicillin (gene ampR), originally
present on the parental form pUC18.
Hybridizations of Southern blots and amplifications through
chain reaction of DNA-polymerase (PCR) were presented to
demonstrate the integration of DNA fragment on vegetable
genome, the number of gene copies, the presence, or absence
of other DNA elements, and the location of transgene. The
results presented corroborate to the statements of the
proponent that one transgenic copy was integrated to a long
arm of chromosome 8 of the corn originally transformed,
and, part of it, transferred to the progenies in
hemizygote, initially, and hemizygote on final versions of
parental lineages for the production of hybrids. The
location of the insertion was defined by linking molecular
markers of RFLP type (polymorphisms as big as fragments of
DNA generated by hydrolysis with endo-nucleases of
restrictions). These essays demonstrated, also, the
presence of transgenes cry1A(b), pat, and of the origin of
replication of pUC18. Finally, such analysis allow for the
conclusion that none of the lineages or hybrids derived
from the initial event Bt11 contain gene ampR.
Results presented by the proponent regarding the analysis
of the presence of cry1A(b) and pat, as well as the pattern
of resistance to glufosinate and to S. frugiperda worms,
have demonstrated that genes Btk and pat are closely
linked, and that both are inherited as loci simple
dominants on Bt11 corn lineage. The segregation data match
the Mendelian pattern on the proportion 3:1 for
heterozygote progeny.
The proof of the presence of recombinant proteins in
different vegetables tissues was executed through imunodetection
of Cry1A(b) and PAT. For the first protein,
higher levels were observed on leaves, with 27 to 33 μg/g
of fresh tissue. Levels 5 to 10 times lower were observed
in straw tissues, stem and grains. For PAT, the amount
described on leaves is around 44 ng/g of fresh tissue. Half
of this value was found in panicles, and 10 times less in
style-stigmas.
IV. Aspects Related to Human and Animals’ Health
The evaluation of foods safety derived from genetically
modified raw material is based on risk analysis, scientific
methodology that encompasses the phases of evaluation,
management and risk communication. On the risk evaluation
phase one looks for the qualitative and quantitative
characterization of potential adverse effects, having as
base the concept of substantial equivalence for the
identification of eventual differences between the new food
and its conventional correspondent. The Principle of
Substantial Equivalence is key concept on the evaluation
process of innocuousness of foods coming from new
technologies (27).
To evaluate safety of genetically modified food raw
material, or its equivalence to conventional food, it is
recommended that four main elements are analyzed, more
specifically: (1) parental variety, that is, the plant that
originated the new genetically modified raw material; (2)
the transformation process, including the characterization
of the construction used, and of the resulting event; (3)
the product of the inserted gene, and the potential of
toxicity and allergenicity, and, finally; (4) the
composition of the new variety deriving from the genetic
transformation. The group of data of these analyses should
allow for the identification and characterization of the
potential adverse effects associated to the new raw
material consume, subsidizing the phases of management and
risk communication.
According to the petitioner, corn Bt11 derives from the
transformation of common Zea mays, a species profoundly
characterized, and about which there is solid safety
background for human consume. Information about identity,
origin and chemical composition have been reported, being
attached to the process publication copy that provides
abundant data regarding its composition, highlighting the
naturally observed variations on the presence of
nutrients(73). The characterization of Bt11 corn, and its
products of expression were extensively analyzed, according
to item III of this technical opinion.
The state of the art on the evaluation of toxicity
preconizes the use of essays of animal experimentation, as
scientific form of qualitative and quantitative
characterization of potential adverse effects to human
health caused by the exposition to environmental
intoxicating substance, or present in foods. Thus, whenever
viable, toxicological essays of xenobiotic in
experimentation animals are executed, administrated them
through exposition via that allow extrapolating the results
observed in animals to humans. This extrapolation allows to
establish IDA (Acceptable Daily Ingestion), or Reference
Dose that means the dose of this substance to which an
individual may expose himself daily without observing
negative effects deriving form such exposition.
Thus, the study of protein Btk of corn Bt11 was conducted
through acute oral via in rats, besides the digestibility
essays. On the essay of simulated digestion, it was
observed that the half-life of the protein is inferior to
30 seconds on the gastric system, and that, in the
intestines, the complete chain protein is converted into
the central fragment resistant to tripsine. The toxicity
study through acute oral via was conducted in rats, and no
harmful effects were observed on any of the evaluated
doses, being 4, 000 mg/kg of body weight the highest dose
tested, which is considered the NOEL of the essay, that is,
the highest dose in which no harmful effects are observed,
estimating thus, DL50 as being superior to 4,000 mg/kg of
body weight. Toxicological classification tables consider
low toxicity doses over 2,000 mg/kg of body weight that do
not provoke harmful effects on evaluated animals under
adequate experimental conditions.
One can conclude that the absence of effects in this essay
was related to the low potential of absorption of protein
demonstrated on the study of in vitro digestibility, where
one could observe its rapid degradation in the gastric
fluid of mammals, with less than 4% of activity after two
minutes. This essay demonstrated the stability of the
protein for 19 hours in the intestinal fluid.
The results show that genetically modified corn on the
concentration of up to 4,000 mg/kg was incapable of
producing acute toxic effects in rats, and that on the
concentration of 11% to 33% on the diet (11g to 33 g/kg of
body weight) it was incapable of producing intoxication
signs in rats fed for 90 days. The Codes Alimentarius of
FAO/WHO(28) uses the following formula for the calculation
of IDA = NOEL/FS
where:
· IDA is the biggest amount in mg/kg of a chemical
substance that can be ingested per day by the human
being, during his whole life, and that does not cause
any harm;
· NOEL is the biggest dose of a chemical substance in
mg/kg that, if used, does not produce toxic effects
on animal species most sensitive to it;
· FS is the safety factor, usually equals to 100 (two
order factors 10: the first considering the human
being 10 times more sensible than the most sensible
animal species studied, and the second considering
the individual variability within the human species).
In this sense, once the biggest amount of genetically
modified corn used in toxicity essays (33,000 mg/kg/day in
sub-chronic essay in the rat) did not produce toxic
effects, and, considering the impossibility of
administrating a bigger amount per day on rats, one can
conclude that it is impossible to calculate the NOEL value.
In fact, a rat does not ingest 10g/100g per day of body
weight of ration, according to Harkness and Wagner’s
description(36), being impossible to feed it with bigger
amount of the product without causing malnutrition due to
lack of other normal ration components. Thus, one can
understand why there are no IDA values for genetically
modified corn. In other words, the level of its possible
toxicity, if it exists, is way beyond the maximum amount
ingested by any human or animal that, in practice, one can
affirm for its absolute innocuousness.
Brake and collaborators(9) compare the nutritional effects
of Bt corn to non modified corn in chicken for slaughter.
The results showed that the administration of genetically
modified corn during 35 days did not interfere with the
gain of weight, or with the digestibility characteristics
of proteins ingested by the chickens. These results were
confirmed, among others, by Taylor et al.(68). Folmer and
collaborators(31) compare the nutritional effects of corn
Bt with non-modified corn in cattle for slaughter and
concluded that the administration of corn Bt did not modify
any parameters that indicate food efficiency, or of gain of
weight of the treated animals in relation to those of the
control group. Sanden and collaborators(58), during a long
term study (8 months) in salmons, reported the lack of
alterations in the body development, and on tissues of the
fish stomach and intestines.
Proteins Cry and PAT have high molecular weight, 65kD and
30 kD, respectively. So, they are not volatile, nor
absorbed by the epidermis, and, for these reasons, it is
not justifiable to evaluate the toxicity of these proteins
through inhaling or dermis via. Additionally, the toxicproteins
safety of B. thuringiensis have been proved since
the 60’s, with the use of microbial insecticides based on
Bt (62, 63, 64), even in organic cultures.
The allergenic potential of proteins Cry 1Ab and PAT was
investigated using various criteria, including homology of
the sequence of amino acids with allergenic known at the
data banks of public domain (Genpept, Swissprot, PIR
protein), and no homology was detected(42). On the contrary
of known proteic allergenic, studies have demonstrated that
proteins Cry1Ab were rapidly inactivated when subjected to
simulated gastric fluids of mammals. Similarly, it was
noted that protein PAT was rapidly digested in conditions
that reproduce human digestion.
Okunuki and collaborators(53) showed that the degradation
of protein Cry1Ab, after being heated is very fast, and,
considering its digestibility in human gastric fluids, they
suggested that it should present no allergenic potential,
or extremely low one. Batista et al.(5) tested the
allergenicity of genetically modified soy and corn in
sensitized individuals, comparing it to the one produced by
conventional seeds in the same individuals, and showed that
the genetically modified products are safe regarding the
allergenic potential. Nakajima and collaborators(50)
confirm the previous data when they reported the lack of
meaningful levels of specific IgE against Cry1Ab in
patients' serum with food allergy.
Chowdhury and collaborators(16) studied the destination of
intrinsic genes (of corn itself), and recombinants in bullcalves
fed with Bt11 corn resistant to insects, and noted
the presence of intrinsic and recombinant genes in the
fluid of rumen, and in the content of the rectus in the
period between five to eighteen hours after being fed.
However, recombinant genes were never found in blood cells,
or guts, and muscles. Phipps and collaborators(54), in
similar work, but with bull-calves fed with ration
containing genetically modified soy (gene cp4-epsps) and Bt
corn (gene cry1Ab), found fragments of transgenes in the
rumen, and in the duodenal digest. There were no traces of
transgene in feces, in the blood, or in the animals’ milk.
Aeschbacher et al.(1), in experiments executed with chicken
fed with hybrid Bt corn, did not find any fragment of
transgene in the tissues of the muscles, liver, spleen,
other organs, flesh or eggs.
A little discussed theme, but with positive impact over
human and animal’s health is the possibility of having the
improvement on grains quality, due to the introduction of
Cry toxin in corn. Due to the greater protection of plants
to insects attack, and, especially, of Bt11 corn spikes,
rotten grains and spikes are extremely reduced when
compared to untransformed plants. As a consequence, they
diminish the toxins of fungus origin on the grains,
reducing the possibility of humans and animals’
intoxication. Munkvold et al’s(49), and Clements et
al’s(18) works in 2003 concluded that B11 corn presented
reduction on the concentration of fungus in the grains.
Between grains and sawdust, the parameters evaluated
presented a similar profile, and within the amplitude used
as reference by the International Life Sciences Institute
Crop Composition(40). The parameter of total grease
percentage by dry weight of grains of Bt11 corn was
superior, when compared to the other treatments. However,
the fatty acids levels were individually presented within
the amplitude published by ILSI(40). The results obtained
indicated that no meaningful unintentional biological
change occurred on the composition, or on the nutritive
value of the grain and of Bt11 corn sawdust, due to the
expression of transgenes cry1A(b) and pat, suggesting,
then, that Bt11 corn is substantially equivalent in
nutritive compositions to the respective isogenic hybrid
not genetically modified and commercial hybrids of corn.
From the analysis of residues (proteins) eventually present
in food coming from Bt11 corn to be provided to animals and
to human beings, one can conclude that none of them have
cancer, teratogenic or genotoxic potential. In fact, these
proteins do not have any structural similarity with primary
or secondary carcinogens, and have no conditions of
connecting to human DNA.(15). Finally, the lack of acute,
or sub-chronic effects produced by genetically modified
corn eliminates, also, any possibility of late
neurotoxicity. This toxic effect is exclusive of
organophosphorate plaguecide, and does not have any
relation to possible residues of Bt11 corn.
Before the foregoing, it is relevant to remind that
allergenic or toxic effects coming from genetically
modified plants were not found. Genetically modified
proteins are degraded by digestion of food, by gastric
fluids, and by bacteria present in the gastrointestinal
treat of human beings and animals.
V. Environmental and Agronomic Aspects
Corn plants are allogamous and annual, of crossed
fecundation and widely pollinated with the help of the
wind, insects, gravity and other agents. The introduction
of genic elements characterized in Bt11 event did not alter
the reproductive characteristics of the plant. Therefore,
the same chances of crossed fecundation that occurs between
hybrids, and not genetically modified lineages of corn,
will occur between plants of Bt11 event, and other corn
plants. In Brazil there are no parental species of corn in
natural distribution. However, there are populations of
creoles corn that can be crossed with genetically modified
corns, in case they are planted in the vicinities.
The risk of passing the transgenes to other individuals in
nature, and its consequences, mostly in biodiversity is,
without any doubt, one of the direct effects that have
called the most attention in case of transgenic. The gene
flow may be horizontal, when the exchange of genetic
information happens between animals of different species,
genetically distant, or vertical when the passage of
genetic information occurs between individuals of the same
species.
The horizontal gene flow between Bt and other species, even
those very related, have almost null probability of
occurrence. Sylvan species related to corn do not naturally
occur in Brazil. Siqueira and collaborators (66) and
Nielsen et al. (51) discuss the possibility of Bt gene of
transgenic plant passing to other microorganisms of the
soil. The conclusion is that the probability is very
remote. Once B. thuringiensis is a soil microorganism, the
exposition of live organisms and of the environment to
these bacteria, or to any element extracted from it is an
event that occurs abundantly in nature, not resulting in
meaningful risk for soil micro biota. It would be much more
plausible for this gene to pass from B. thuringiensis to
other micro-organisms.
The vertical gene flow, at first, has no consequence
because most agriculturists do not reuse the collected
grains as seeds. The hybrid seeds of F1 generation are
acquired every year. However, there is a small contingent
of agriculturists of subsistence that keep creoles
varieties. Nodari and Guerra(52) argue that the diversity
of agricultural species composed of creoles cultivars of
corn may be threatened by transgenic. However, it is
possible to keep these cultivars, for hybrid corn has been
intensively cultivated in Brazil for many decades in the
same regions in which most of the creoles cultivars are
concentrated and the latter have been kept. On the other
hand, even if gene flow occurs between plants of Bt11 corn,
and creoles varieties, it is not expected difference of
genic flow in relation to any other allele that exists in
plants. Discussion in this regard is presented by Ramalho
and Silva(59). In sum, gene or allele will only remain in
the population if the genic flow is continuous, with a
relatively high frequency, and if there is any adaptation
advantage. Additionally, the characteristics introduced
into event Bt11 would not bring potentially damaging
consequences to human, animal’s health, or to the
environment, due to the considerations made previously, and
to the background of safe use in other countries for more
than 10 years(12). However, it is necessary to emphasize
that the coexistence between conventional cultivars of corn
(improved or creoles) and transgenic cultivars of corns is
possible from the agronomic point of view(11,46), and one
should note the disposition on Normative Resolution No.4 of
CTNBio. It is also important to remember that most of
indigenous races, creoles populations, ancient and recent
cultivars, as well as exotic cultivars of corn are
preserved in Brazil by EMBRAPA, as well as in various
institutes of germoplasm preservation in the world.
Classical analysis of the genetics presented by the
proponent has demonstrated that there is no possibility of
distinction between the pollen of Bt11 corn and the pollens
of non-transgenic corns. The results pointed out to the
fact that heterozygote corn plants for genes cry1A(b) and
pat do not produce progenies excess in crossing-test,
concluding that Bt11 corn pollen is not more competitive,
or efficient in fertilization than the conventional pollen.
Comparing the pollen concentrations to 1m of source culture
under low to moderate winds, it was estimated that,
approximately, 2% of pollen is noted at 60m, 1.1% at 200m,
and 0.75-0.5% at 500m of distance. At 10 m of a field, in
average, the number of pollen grains per area unit is ten
times smaller than the one observed at 1m from the border.
Therefore, if the established distances of separation
developed for the production of corn seeds are observed, it
is expected that the pollen transfer to the adjacent
varieties are minimized, being improbable the presence of
genetic materials with resistance to insects.
Seeds dispersion is easily controlled, once corn
domestication eliminated the ancestral mechanisms of seeds
dispersion, and the pollen movement is the only effective
mean of corn plants genes escape, thus, in face of the
nature of grains, cobs and corn plants, this vegetable
survival is limited to the plantation, and harvest cycle
made by human being, since it is totally dependent on him
for the seeds to germinate after being thrashed. The
different vegetable tissues and organs do not have
proliferation capacity, being restricted to seeds firmly
stuck to the spikes, and protected by straw, that is, only
human activity can remove the seeds from the spikes, and
guarantee the survival of the vegetable, cycle to cycle.
Thus, corn plants are not invasive plants, and their
control is easily executed on crops where cultures
rotations are conventional practices, with eventual arise
of voluntary, or spontaneous plants derived from seeds lost
during harvest. In the Brazilian environment, where native
species sexually compatible with corn do not occur, or are
not known, the risk of Bt11 corn execute, or promote the
invasion of uncultivated and cultivated areas does not
exist.
With expected effects of transgenic expression, an
incomplete version of protein Cry1A(b) is expressed on
vegetable tissues. Lepidopterus insects S. frugiperda , H.
zea and D. saccharalis are particularly susceptible to the
action of this class of á-endotoxins, for they have
digestive treat with alkaline pH, what promotes the
solubilization of proteic crystals, and the intestinal
receptors specific to them. This endotoxin ingestion by
worms with alkaline digestive environment will promote the
death of the insects through the interaction of the protein
with intestinal receptors of cellular surface, promoting
the opening of the pores, and the invasion of
microorganisms of the intestinal treat. Thus, the insects’
death derives from the osmotic unbalance promoted by the
toxin, and by septicemia deriving from the invasion of the
intestinal flora by microorganisms(10).
One of the advantages of transgenic plants resistant to
insects expressing genes that codify á-endotoxins, or the
microbial preparations, when compared to chemical
insecticides, is the high specificity to target-species. In
fact, no differences were observed among populations of
Dermaptera: Forficulidae, Coleopteran: Anthocoridae,
Carabidae, Cincidelidae and Araneae. In relation to eggs
parasitism of H. zea by Trichogramma sp. (Hymenoptera:
Trochogrammatidae), no meaningful differences were observed
either when compared to Bt11 plants with their isogenic
lineage not genetically modified(30). The results reinforce
observations made in other countries and cultures where
field studies showed that the abundance and activity of
untargeted insects (predators and parasitoids) were similar
when plants genetically modified with Bt were compared to
non-genetically modified plants. In contrast, crops whose
control is made through chemical methods, negative effects
are normally observed on the biological control of plagueinsects.
Before the foregoing, one can conclude that the
use of Bt plants, and the consequent reduction on the
applications of insecticides tend to favor the presence of
predators insects and parasitoids of plague-insects(57).
In relation to target-insects, this event was tolerant to
the attack of H.zea, almost no damage occurred on spikes,
and to the attack of S. frugiperda. In severe infestation
conditions of S. frugiperda, the proponent demonstrated
that hybrids of Bt11 corn presented productivity extremely
higher than that the one of its non-transgenic isogenies.
In fields experiments carried out in 2000 in Uberlândia,
MG, Bt11 corn also showed a noted effect over Mocis latipes
(plague of Lepidoptera order that feeds from leaves).
According to analysis of factorial variance of the data
presented by the proponent, no difference was observed
between the hybrids derived from original elite lineages
and the derivatives of Bt11 converted lineages selected for
the aspects of productivity, humidity on harvest, putting
roots on the ground, spikes height, plants height, and
thermal units for adornment, or dehiscence of pollen
grains. However, meaningful differences were described
between the original elite-lineages, and the conversions
Bt11 for the characteristics of putting the stem on the
ground, and integrity note. Bt11 corn presented smaller
stem breakage than the non-transgenic hybrids, due to the
fact that the former is less susceptible to damages on the
leaves and on the stem, due to the smaller incidence of
plague-lepidopteron. In relation to differences between
grains produced by Bt11 corn, and by equivalent
conventional corn, the analysis results of spectroscopy of
near infra-red (NIRS) have demonstrated that there are no
differenced in relation to non-transgenic grains for
density, weight of 100 grains, grains size, amid
percentage, protein percentage, oil percentage, and fiber
percentage.
Agronomic parameters, and the efficacy on plaguelepidopteron
control of Bt11 corn hybrids were compared to
isogenic lineages in essays conducted in 5 places:
Uberlândia-MG, Ituiutaba-MG, Iraí de Minas-MG, Campo
Mourão-PR and Pinhalzinho-SC, in the agricultural harvest
of 2005/06. Plants structure, spikes insertion height, male
and female flowering date, note for diseases, percentage of
erect plants, kind of grains, gains color, humidity
content, profit, and rancid grains, were the parameters
studied on the agronomic evaluations. For the study of
efficacy of the event Bt11 in the control of lepidopteronplague
damage of S. frugiperda, of D. saccharalis, and of
H. zea were evaluated. Bt11 hybrids were efficient for the
control of the evaluated lepidopteron-plague, as well as
superior for the agronomic parameters grains profit and
rancid grains. According to presented information the
favorable differential of performance was mainly related to
the efficient protection against the attack of the plagues
studied. For the other evaluated agronomic parameters Bt11
hybrids presented performance statistically equals to the
respective non GM isogenic hybrids. These results confirm
the equivalence of agronomic performance between Bt11
hybrids, and the non GM isogenies in cultivation conditions
of the culture in Brazil.
In Brazil nowadays, there is indiscriminate use of
insecticides, and even mixture of chemical products, trying
to control insects, especially S. frugiperda. With the
massive application of these chemical products, an
agricultural desert is created in certain regions of
Brazil, for the natural enemies of plagues are the first to
be eliminated. The frequent application of chemical
insecticides contributes for the degradation of the
environment, environmental pollution and break of all the
ecosystem in corn culture, and even in other cultures in
rotation. With the adoption of genetically modified plants
resistant to insects, the reduction of insecticides has
been considerable in countries that have adopted the
technology for more than ten years. For example, in the
United States, producers have obtained reduction of more
than 8,000 tons of insecticide active ingredient only in
2001(14,34,33). In China, the applications of insecticides
were reduced in an average of 67%, and the reduction in
volumes of insecticide active ingredients was reduced in
80%(38). In South Africa the reductions were around
66%(41). Before the foregoing, one can consider that the
use of Bt technology in Brazil may contribute for the
reduction of the use of insecticides and consequently,
reduce impacts of use of these agro toxic in the
environment, in the human and animal’s health. What’s more,
the use of Bt technology may have positive impact on the
preservation of populations of untargeted organisms and
benefic insects, facilitating the integrated handling of
crop plagues(69, 37,6). Additionally, the adoption of
technologies that reduce pulverization of chemical products
in crops may favor acquiring secondary benefits, such as
the reduction of use of raw-material on the production of
agro toxics, on the conservation of fuel used to produce,
distribute and apply such agro toxic, and for the
elimination of use necessity and discard of agro toxic
cartons(44).
VI. Restriction to the use of GMO and its derivatives:
Studies presented by the petitioner demonstrated that there
was no meaningful difference between the hybrids of corn
derived from unmodified lineages and Bt11 corn in relation
to agronomic characteristics, such as productivity, harvest
humidity, putting the root on the ground, spike height,
plant height, and others. Besides, there were no meaningful
differences in the reproduction way, dissemination or
capacity of survival of the genetically modified corn in
relation to lineages of unmodified corn. All the evidences
presented in the process, and in bibliographic references
such as Schuler et al.(60), of Maagd et al. (22), Candas
and collaborators(13), Brookes et al. (11), Broderick et
al. (10), Sanden et al. (58), Okuniki et al. (53), among
others, confirm the risk level of the transgenic variety as
equivalent to the non-transgenic varieties in face of the
soil micro flora, to untargeted vertebrate and invertebrate
animals, as well as to other vegetables, and to human and
animal health. Thus, the cultivation and consume of Bt11
corn are not potentially causing meaningful degradation of
the environment, or risks to human and animal’s health. For
these reasons, there is no restriction to the use of this
corn, or its derivatives.
After ten years of use in different countries, no problem
was detected for the human and animal’s health, or for the
environment that may be attributed to transgenic corn. It
is necessary to emphasize that the lack of negative effects
resulting from transgenic plants cultivation of corn does
not mean that they cannot happen. Zero risk to absolute
safety does not exist in the biological world, however,
there already exists an accumulation of trustworthy
scientific information , and a safe background of ten years
use that allows us to affirm that Bt11 corn is as safe as
its conventional versions. Thus, the petitioner should
conduct monitoring after the commercial release on the
terms of Normative Resolution No. 3 of CTNBio.
The vertical gene flow for local varieties (called creoles
corns) of open pollination is possible, and presents the
same risk caused by commercial genotypes available in the
market (80% of conventional corn planted in Brazil come
from commercial seeds that went through a process of
genetic improvement). The coexistence of conventional corns
cultivations (improved or creoles), and transgenic
cultivations of corns is possible from the agronomic point
of view(11, 46), and should follow the disposition on
Normative Resolution No. 4 of CTNBio.
VII. Considerations about particularities of different
regions of the Country (subsidies to the inspections
organs):
In accordance with what is established on art. 1 of Law
11.460, of March 21st, 2007, “it is vetoed the research and
cultivation of organisms genetically modified on indigenous
lands and areas of conservation units”.
VIII. Conclusion
Considering that Bt11 corn derives from the transformation
of common corn Z. mays, species profoundly characterized,
and about which there is solid safety background for human
and animal consume, and that the transformation process
gave place to the insertion of a sole copy of the fragment
of DNA containing the genetic constructions with genes pat
and Btk.
Considering that the safety of corn containing gene pat was
exhaustively analyzed by CTNBio on process
01200.005154/1998-36, and, moreover, that on Technical
Opinion 987/2007 all aspects related to biosafety of
Liberty Link corn were approached.
Considering also that:
1. Corn is the species that reached the highest degree of
domestication among cultivated plants, being able to
survive in nature without human intervention.
2. There is no sylvan species in Brazil with which corn can
be crossed, since the closer sylvan species to corn is
teosinte, found in Mexico and in some places in Central
America, where it can be crossed with corn cultivated in
production fields.
3. Protein Cry1Ab was detected in low levels of analyzed
tissues, and presented great susceptibility to digestion in
simulations of gastric fluids, not demonstrating acute
toxicity in mammals, or similarity with known allergens.
4. Due to the greater protection of plants to insects’
attack, particularly, of Bt11 corn spikes, rotten grains
and spikes are meaningfully reduced when compared to
untransformed plants, consequently, there is reduction of
toxins of fungus origins in grains, diminishing the
possibility of intoxication of humans and animals.
5. No unintentional meaningful biological change occurred
on the composition, or on the nutritious value of the grain
and of the Bt11 corn sawdust, due to Cry1A(b) and pat
transgene expression, suggesting, then, that Bt11 corn is
substantially equivalent in nutritious composition to the
respective isogenic hybrid not genetically modified, and to
commercial corn hybrids.
6. DNA molecule is a natural food component, not presenting
any evidence that such molecule may have adverse effect to
men when ingested in food in acceptable quantities (no
direct toxic effect).
7. There is no evidence that intact genes of plants may be
transferred and functionally integrated to human genome, or
to other mammals exposed to this DNA, or foods manufactured
with these elements(16).
8. The petitioner answered to all the questionings
postulated on Normative Instruction No. 20 of CTNBio, and
none of the questions indicate that this corn may present
adverse effects on human or animal food.
9. There is no risk of Bt11 corn to execute or promote
invasion of uncultivated areas.
10. B. thuringiensis may be considered the biological agent
of greatest potential for the control of forests,
agricultural plague-insects, and vector of diseases, thanks
to the specificity of endotoxins to insects and targetinvertebrate,
and its innocuousness to vertebrates and
environment, including benefic insects and natural enemies,
making this agent a key-component in strategies of
integrated handling of plagues.
11. B. thuringiensis cultures are registered in the
National Health Surveillance Agency – ANVISA under
different formulations for the application in 30 kinds of
vegetable cultures for food use.
12. Bio-pesticides based on toxin are widely used as an
alternative to chemical insecticides in terms of safety to
non-targeted organisms, and when the development of
resistance to chemical insecticides occurs.
13. Meaningful differences were not observed between the
populations of ladybugs, Carabidae, cincidelidae and
spiders, as well as parasitoid of H. zea, Trichogramma sp.,
when plants Bt11 are compared to their isogenic lineage not
genetically modified.
14. One of the advantages of transgenic plants resistant to
insects expressing genes that codify endotoxins, or
microbial preparations, when compared to chemical
insecticides, is the high specificity to target-species.
15. The use of Bt technology in Brazil may contribute for
the reduction of the use of insecticides, and,
consequently, reduce the impacts of the use of such agro
toxics in the environment, in human and animal’s health,
and it may also indirectly help on the preservation of nontargeted
organisms populations and benefic insects,
facilitating the integrated handling of crop plagues.
16. The use of genetically modified plants resistant to
insects present positive repercussions also in the aspects
related to the acquisition and use of chemical
insecticides, to meaningfully reduce the pollution provoked
by industrial rejects, and by the use of water used on
pulverizations, besides avoiding man, food, rivers and
springs contamination deriving from the use, transportation
and storage of insecticides.
17. The coexistence among cultivations of conventional
corns (improved or creoles) and transgenic cultivations of
corns is possible from the agronomic point of view, and one
should observe the disposition on Normative Resolution No.
4 of CTNBio.
18. Comments, opinions, suggestions and documents resulting
from the Public Hearing that took place on March 20th, 2007
did not present relevant scientific fact, substantiated by
scientific evidences that compromise environmental safety
of human and animals’ health of corn Bt11.
19. Attachment III of Cartagena Protocol about Biosafety
(Decree 5.705, of February 16th, 2006) says that risks
associated to live organisms modified, or to products
derived from them, to wit, benefited materials that have as
origin a live modified organism, containing new detectable
combinations of replicable genetic material obtained
through the use of modern biotechnology, should be
considered on the context of risks presented by the nonmodified
receptors or parental organisms in the probable
receptor environment.
20. The historical use of this transgenic variety in the
world reveals a great accumulation of trustworthy
scientific information that indicate that this variety is
as safe for the environment, and for human and animal
health, as the varieties of hybrid corns that have been
being used.
21. After ten years of use in different countries, no
problem was detected for human, animal’s health, or to the
environment that may be attributed to transgenic corns. It
is necessary to emphasize that the lack of negative effects
resulting of corn transgenic plants does not mean that they
may not happen. Zero risk and absolute safety does not
exist in the biologic world, although there already exist
an accumulation of trustworthy scientific information and a
safe background of ten years of use that allows us to
declare that corn Bt11 is as safe as conventional versions.
Thus, the petitioner should conduct monitoring of postcommercial
release on the terms Normative Resolution No. 3
of CTNBio.
Before the foregoing, and considering the international
criteria accepted on the process of risk analysis of
genetically modified raw-material, it is possible to
conclude that Bt11 corn, derived from MON810 lineage, is as
safe as its conventional equivalent.
CTNBio thinks that the cultivation and consume in
commercial scale of Bt11 corn are activities that do not
potentially cause meaningful degradation of the environment
or aggravations to human and animal health. The use
restrictions of the GMO in analysis and its derivatives are
conditioned to disposition on Normative Resolution No. 03
and Normative Resolution No. 04 of CTNBio.
VIII – Bibliographic References
1. AESCHBACHER, K.; MESSIKOMMER, R.; MEILE, L; WENK, C.
2005. Bt176 corn in poultry nutrition: physiological
characteristics and fate of recombinant plant DNA in
chickens. Poultry Sci. 84: 385-394.
2. ALVES FILHO, J.P. 2001. Agrotóxicos e Agenda 21:sinais e
desafios da transição para uma agricultura sustentável.In:
II SINTAG Anais. II Simpósio Internacional de Tecnologia de
Aplicação de Agrotóxicos: eficiência, Economia e
Preservação da Saúde Humana e do Ambiente, Jundiaí, SP,
17/07/2001 a 20/07/2001.
3. ANVISA. 2006.
http://www.anvisa.gov.br/toxicologia/monografias/b01.pdf.
Acesso em 15/10/2006.
4. BAHIA FILHO, A.F.C.; GARCIA, J.C. 2000. Análise e
Avaliação do mercado brasileiro de sementes de milho. In:
UDRY, C.V.; DUARTE, W.F. (Org.) Uma história brasileira do
milho: o valor de recursos genéticos. Brasília: Paralelo
15, 167-172.
5. BATISTA, R.; NUNES, B.; CARMO, M.; CARDOSO, C.; JOSÉ,
H.; DE ALMEIDA, A.; MANIQUE, A.; BENTO, L.; RICARDO, C.;
OLIVEIRA, M. 2005. Lack of detectable allergenicity of
transgenic maize and soya samples. J. allergy Clin.
Immunol. 116:403-10.
6. BENEDICT, J; ALTMAN, D. 2001. Commercialization of
transgenic cotton expressing insecticidal crystal protein.
In: JENKINS, J.; SAHA, S. (eds.). Genetic improvement of
cotton: emerging technologies. Enfield: Science Publishers,
137-201.
7. BENSASSON, D.; BOORE, J.L.; NIELSEN, K.M. 2004. Genes
without frontiers? Heredity, 92: 483-489.
8. BRADFORD, K.J.; DEYNZE, A.V.; GUTTERSON, N.; PARROTT,
W.; STRAUSS, S.H. 2005. Regulating transgenic crops
sensibly: lessons from plant breeding, biotechnology and
genomics. Nature Biotech. 23: 439-444.
9. BRAKE, J.; FAUST, M.A.; STEIN, J. 2003. Evaluation of
transgenic event Bt11 hybrid corn in broiler chickens.
Poultry. Sci. 82:551-559.
10. BRODERICK, N.A.; RAFFA, K.F.; HANDELSMAN, J. 2006.
Midgut bacteria required for Bacillus thuringiensis
insecticidal activity. Proc. Natl. Acad. Sci. USA 103:
15196-15199.
11. BROOKES, G.; BARFOOT, P.; MELÉ, E.; MESSEGUER, J.;
BÉNÉTRIX, F. BLOC, D.; FOUEILLASSAR, X, FABIÉ,A.;
POEYDOMENGE, C.2004. Genetically modified maize; pollen
movement and crop co-existence. Dorchester, UK: PG
Economics, 20pp.
(www.pgeconomics.co.uk/pdf/Maizepollennov2004final.pdf)
12. BROOKES, G.; BARFOOT, P.2006. Global Impact of Biotech
crops; Socio-Economic and Environmental Effects in the
First Ten Years of Commercial Use. AgBioForum 9: 139-151.
13. CANDAS, M.; LOSEVA, O.; OPPERT, B.; KOSARAJU, P.; BULLA
JUNIOR, L.A. 2003. Insect resistance to Bacillus
thuringiensis: alterations in the Indian meal moth gut
proteome. Molec. Cel. Proteomics 2.1: 19-28.
14. CARPENTER, J.; FELSOT, A; GOODE, T.; HAMMING, M.;
ONSTAD, D.; SANKULA, S. 2002. Comparative environmental
impact of biotechnology-derived and traditional soybean,
corn, and cotton crops (CAST: 1-189). Ames, IA: Council for
Agricultural Science and Technology.
15. CASARETTE, L.J.; DOULL, J. 1975. Toxicology: the basis
science of poisons. New York: Mcmillian Publishing Co., p.
313-378.
16. CHOWDHURY, E.H.; KURIBARA, H.; HINO, A.; SULTANA, P.;
MIKAMI, O.; SHIMADA, N.; GURUGE, K.S.; SAITO, M.; NAKAJIMA,
Y. 2003. Detection of corn intrinsic and recombinant DNA
fragments and Cry1Ab protein in the gastrointestinal
contents of pigs fed genetically modified corn Bt11. J.
Anim. Sci. 81: 2546-2551.
17. CHRISTOU, P.; CAPELL,T.; KOHLI, A.; GATEHOUSE, J.A.;
GATEHOUSE, A.M.R. 2006. Recent developments and future
prospects in insect pest control in transgenic crops.
Trends plant Sci. 11: 302-308.
18. CLEMENTS, M.J.; CAMPBELL, K.W.; MARAGOS, C.M.; PILCHER,
C.; HEADRICK, J.M.; PATAKY, J.K.; WHITE,D.G. 2003.
Influence of Cry1Ab protein and hybrid genotype on
fumonisin contamination and Fusarium ear rot of corn. Crop
Sci. 44: 1283-1293.
19. CONAB. Milho total (1a e 2a safra) Brasil – Série
Histórica de área plantada: safra 1976-77 a 2006-07.
HTTP://www.conab.gov.br/conabweb/download/safra/MilhoTotal
SerieHist.xls
20. CRICKMORE, N. 2007. Bacillus thuringiensis Toxin
Nomenclature.
http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/
21. CRUZ, I. FIGUEIREDO, M.L.C.; OLIVEIRA, A.C.;
VASCONCELOS, C.A. 1999. Damage of Spodoptera frugiperda
(Smith) in different maize genotypes cultivated in soil
under three levels of aluminum saturation. International
Journal of Pest Management 45:293-296.
22. DE MAAGD, R.A.; BRAVO, A.; CRICKMORE, N. 2001. How
Bacillus thuringiensis has evolved specific toxins to
colonize insect world. Trends Genet. 17: 193-199.
23. DE MAAGD, R.A.; BRAVO, A.; CRICKMORE, N. SCHNEPF, H.E.
2003. Structure diversity and evolution of protein toxins
from spore-forming entomopathogenic bacteria.
Ann.Rev.Gent.37: 409-433.
24. EDGE, J.M.; BENEDICT, J.H.; CARROLL, J.P.; REDING, H.K.
2001. Bolgard cotton: an assessment of global economic,
environmental and social benefits. J. Cotton sci %: 121-
136.
25. FAO/WHO – Food and Agriculture Organization of the
United Nations. 2000. Safety aspects of genetically
Modified Foods of Plant Origin. Report of a Joint FAO/WHO
Expert Consultation on Foods Derived from Biotechnology, 29
May – 2 June, 2000. World Health Organization, WHO
Headquarters, Geneva, Switzerland. 35pp.
http://www.org.int/foodsafety/publications/biotech/en/ec_Ju
ne2000_en.pdf
26. FAO/WHO – Food and Agriculture Organization of the
United Nations. 2000. Grassland Index Zea mays L.
(http://www.fao.
Org/WAICENT/agricult/agp/agpc/doc/gbase/data/pf000342.htm)
27. FAO/WHO – Organización de las Naciones Unidas para la
Agricultura y la Alimentación/ Organización Mundial de La
Salud. 2004. Codex Alimentarius: Alimentos obtenidos por
medios biotecnológicos. Roma: FAO, 57pp.
28. FAO/WHO. 2006. Evaluation of certain veterinary drug
residues in food. Report of the 66th Meeting of the Joint
Expert Committee on Food Addictives (JECFA). P.9-12.
29. FAO. 2007. FAOSTAT. Disponível em:
http://faostat.fao.org/site/340/default.aspx.
30. FERNANDES, O.A.; FARIA, M.; MARTINELLI, S.; SCHIMIDT,
F.; CARVALHO, V.F.; MORO, G. 20007. Short-Term assessment
of Bt on non-target arthropods in Brazil. Sci. Agric. 64:
249-255.
31. FOLMER, J.D.; GRANT, R.J.; MILTON, C.T.; BECK, J. 2002.
Utilization of Bt corn residues by grazing beef steers and
Bt corn silage and grain by growing beef cattle and
lactating dairy cows. J. Anim. Sci. 80: 1352-1361.
32. FORLANI, G.; OBOJSKA, A.; BERLICKI, L. ; KAFARSKI, P.
2006. Phosphinothricin analogues as inhibitors of plant
glutamine synthetases. J. Agric.Food Chem. 54:796-802.
33. GIANESSI, L.; SILVERS, C.; SANKULA, S.; CARPENTER, J.A.
2002. Plant biotechnology: current and potential impact for
improving pest management in U.S. agriculture – analysis of
40 case studies (executive summary). Washington, DC:
national Center for Food and Agricultural Policy.
http://www.ncfap.org/40CaseStudies/NCFAB%20Exec%20Sum.pdf
34. GÓMEZ, I.; PARDO-LÓPEZ, L.; MUÑOZ-GARAY, C.; FERNANDEZ,
L.E.; PÉREZ, C.; SÁNCHEZ, J.; SOBERÓN, M.; BRAVO, A. 2007.
Role of receptor interaction in the mode of action of
insecticidal Cry and Cyt toxins produced by Bacillus
thuringiensis. Peptides 28: 169-173.
35. GURR, S.J.; RUSHTON, P.J. 2005. Engineering plants with
increased disease resistance: how are we going to express
it? Trends in Biotechnol. 23: 283-290.
36. HARKNESS, J.E.; WAGNER, J.E. 1993. Biologia e Clínica
de Coelhos e Roedores. São Paulo: Roca, 3.ed.p.49.
37. HEAD, G.; FREEMAN, B.; MINA, B.; MOAR, W.; RUBERSO, J.;
TURNIPSEED, S. 2001. Natural enemy abundance in commercial
Bollgard and conventional cotton fields. Proceedings of the
Beltwide Cotton Conference 2: 796-798. Memphis; National
Cotton Council.
38. HUANG, J.; ROZELLE, S.; PRAY, C.; WANG, Q. 2002. Plant
biotechnology in China. Science 295: 674-676.
39. HUANG, Z.; GUAN, C.; GUAN, X. 2004. Cloning,
characterization and expression of a new cry1Ab gene from
Bacillus thuringiensis WB9. Biotechnol. Lett.26: 1557-1561.
40. ILSI. 2004. Nutritional and safety assessment of foods
and feeds nutritionally improved through biotechnology.
Compr. Rev. Food Sci. Food Safety 3: 36-104.
41. ISMAEL, Y.; BENNETT, R.; MORSE, S. 2002. Bt cotton,
pesticides, labour and health: a case study of smallholder
farmers in the makhatini Flats, republic of South Africa.
Paper presented at the 6th International ICABR Conference,
Ravello, Italy.
42. KEEK, P.J.; MITSKY, T.A. 1994. Comparative alignment of
insecticidally-active B.t.k. HD-73 protein (b.t.k. protein)
to known allergenic and toxic proteins using the FAST
algorithm. Monsanto Technical Report MSL – 13643, St.
Louis.
43. KRIEG,A.; LANGENBRUCH, G.A. 1981. Susceptibility of
arthropod species to Bacillus thuringiensis. In: BURGES,
H.D. (Ed.) Microbial control of pests and plant diseases
1970-1980. London: Academic, 837-896.
44. LEONARD, R.; SMITH, R. 2001. IPM and environmental
impacts of bt cotton: a new era of crop protection and
consumer benefits. ISN No. 00401074.
45. LESSARD, P.A.; KULAVEERASINGAM, H.; YORK, G.G.; STRONG,
A.; SINSKEY, A.J. 2002. Manipulating gene expression for
the metabolic engineering of plants. Metabolic Engin. 4:
67-79.
46. MESSEGUER, J.; PEÑAS,G.; BALLESTER, J.; BAS, M.; SERRA,
J.; SALVIA, J.; PALAUDELMÀS, M.; MÉLE, E. 2006. Pollenmediated
gene flow in maize in real situations of
coexistence. Plant Biotechnology Journal. 4:633-645.
47. MONNERAT, R.G.; BRAVO, A. 2000. Proteínas
bioinseticidas produzidas pela bacteria Bacillus
thuringiensis: modo de ação e resistência. In: MELO. In:
MELO, I.S.; AZEVEDO, J.L. (Ed.) 2000. Controle Biológico.
Jaguariúna: Embrapa Meiro Ambiente, 163-200.
48. MOORE, I.; SAMALOVA, M.; KURUP, S. 2006.
Transactivated and chemically inducible gene expression in
plants. Plant J. 45: 651-683.
49. MUNKVOLD, G.P.; HELLMICH, R.L.; RICE, L.G. 1999.
Comparison of fumonisin concentration in kernels of
transgenic Bt maize hybrids and nontransgenic hybrids.
Plant Dis. 83: 130-138.
50. NAKAJIMA, O.; TESHIMA, R.; TAKAGI, K.; OKUNUKI., H.;
SAWADA, J.I. 2006. ELISA method for monitoring human serum
IgE specific for Cry1Ab introduced into genetically
modified corn. Regul. Toxicolog. Pharmacol. 44: 182-188.
51. NIELSEN, K.M.; BONES, A.M.; SMALLA, K.; VAN ELSAS, J.D.
1998 Horizontal gene transfer from transgenic plants to
terrestrial bacteria – a rare event? FEMS Microbiology
Reviews 22, 79-103.
52. NODARI R.O.; GUERRA, M.P. 2001. Assessment of
environmental risks of genetically modified field plants.
Cadernos de Ciência Tecnologia 8: 81-116.
53. OKUNUKI, H.; TESHIMA, R.; SHIGETA, T.; SAKUSHIMA, J.;
AKIYAMA, H.; GODA, Y.; TOYODA, M.; SAWADA, J. 2002,
Increased digestibility of two products in genetically
modified food (CP4-EPSPS and Cry 1Ab) after preheating.
Shokuhin Eiseigaku Zasshi 43: 68-73.
54. PHIPPS, R.H.; DEAVILLE, E.R.; MADDISON, B.C.; Sutton,
J.D.; Beever, D.E.; Givens, D.I. 2003. Detection of
transgenic and endogenous plant DNA in rumen fluid,
duodenal digesta, milk, blood, and feces of lacting Dairy
Cows. J. Dairy Sci. 86: 4070-4078.
55. RAMALHO, M.A.P.; SILVA, N.O. 2004. Fluxo gênico em
plantas. In: MIR, L.; MOREIRA FILHO, C.A. (Eds.) Genômica.
São Paulo: Atheneu. P. 863-884.
56. RODRIGO–SIMÓN A.; DE MAAGD R.A.; AVILLA, C.; BAKKER,
P.L.; MOLTHOFF, J.; GONZÁLEZ-ZAMORA, J.E.; FERRÉ, J. 2006.
Lack of detrimental effects of Bacillus thuringiensis Cry
toxins on the insect predator Chrysoperla carnea: a
toxicological, histopathological, and biochemical analysis.
Appl. Environ. Microbiol. 72: 1595-1603.
57. ROMEIS, J.; MEISSLER, M.; BIGLER, F. 2006. Transgenic
crops expressing Bacillus thuringiensis toxins and
biological control. Nature Biotechnol. 24: 63-71.
58. SANDEN, M.; BERNSTSSEN, M.H.G.; KROGDAHL, D.; HERME, GI.;
MCKELLEP, A-M. 2005. An examination of the intestinal
tract of Atlantic salmon, Salmo salar L., parrfed different
varieties of soy and maize. J. Fish Dis.28: 317-30.
59. SCHNEPF, E.; CRICKMORE, N.; VAN RIE, J.; LERECLUS, D.;
BAUM, J.; FEITELSON, J.; ZEIGLER, D.R.; DEAN, D.H. 1998.
Bacillus thuringiensis and its pesticidal cystal proteins.
Microbiol. And Molec.Biol. rev. 62: 775-806.
60. SCHULER, T.H.; DENHOLM, I.; JOUANIN, L.; CLARK, S.J.;
CLARK, A.J.; POPPY, G.M. 2001. Population-scale laboratory
studies of the effect of transgenic plants on nontarget
insects. Mol. Ecol. 10: 1845-1853.
61.SHELTON, A.M.; ZHAO, J.Z.; ROUSH, R.T. 2002. Economic,
ecological, food safety, and social consequences of the
deployment of Bt transgenic plants. Annu. Rev. Entomol. 47;
845-881.
62. SHIMADA, N.; YONGSOON, K.; MIYAMOTO, K.; YOSHIOKA, M.;
MURATA, H. 2003. Effects of Bacillus thuringiensis Cry1Ab
toxin on mammalian cells. J Vet Med Sci. 65: 187-191.
63. SHIMADA, N.; MIYAMOTO, K.; KANDA, K.; MURATA, H. 2006.
Bacillus thuringiensis insecticidal Cry1Ab toxin does not
affect the membrane integrity of the mammalian intestinal
epithelial cells: an in Vitro study. In Vitro Cell Dev.
Biol. Anim. 42: 45-49.
66. SIQUEIRA, J.O.; TRAMIN, I.C.B.; RAMALHO, M.A.P.;
FONTES, E.M.G. 2004. Interferências no agrossistema e
riscos ambientais de culturas transgênicas tolerantes a
herbicidas e protegidas contra insetos. Cadernos de Ciência
e Tecnologia 21: 11-81.
67. TAN, S.; EVANS, R.; SINGH, B. 2006. Herbicidal
inhibitors of amino acid biosynthesis and herbicidetolerant
crops. Amino Acids 30: 195-204.
68. TAYLOR, M.L.; HARTNELL, G.; NEMETH, M.; KARUNANANDAA,
K.; GEORGE, B. 2005. Comparison of broiler performance when
fed diets containing corn grain with insect-protected (corn
rootworm and European corn borer) and herbicide-tolerant
(glyphosate) traits, control corn, or commercial reference
corn – revisited. Poult.sci. 84: 1893-1899.
69. XIA, J.Y.; CUI, J.J.; MA, L.H.; DONG, S.X.; CUI, X.F.
1999. The role of transgenic Bt cotton in integrated
insect pest management. Acta Gossypii Sim 11: 57-64.
70. YI, G.; SHIN, Y.M.; CHOE, G.; SHIN, B.; KIM, Y.S.; KIM,
K.M. 2007. Production of herbicide-resistant sweet potato
plants transformed with the bar gene. Biotechnol. Lett. 29:
669-675.
71. YU, J.; XIE, R.; TAN, L.; XU, W.; ZENG, S.; CHEN, J.;
TANG, M.; PANG, Y. 2002. Expression of the full length and
3’-spliced cry1Ab gene in the 135-kDa crystal protein minus
derivative of Bacillus thuringiensis subsp. Kyusshuensis.
Curr. Microbiol. 45: 133-138.
72. WAQUIL, J.M.; VILLELA, F.M.F.; FOSTER, J.E. 2002.
Resistência do milho (Zea mays L.) transgênico (Bt) à
lagarta-do-cartucho, Spodoptera frugiperda (Smith)
(Lepidoptera: Noctuidae). Revista Brasileira de Milho e
Sorgo 1 (3): 1-11.
73. WATSON, S.A.; RAMSTAD, P.E. 1987. Corn: chemistry and
technology. St. Paul: American Association of Cereal
Chemist, 605p.
(unreadable signature)
Walter Colli
President of CTNBio
Divergent Vote:
CTNBio’s member, Dr. Rubens Onofre Nodari (Environmental
Permanent Sector Sub-commission) voted contrarily to the
commercial release of Bt11 corn.
The reporter Dr. Fábio Kessler Dal Soglio (Vegetable
Permanent Sector Sub-commission) issued contrary opinion to
this product approval considering the following points:
1. Problems on the characterization of the genetic
transformation event;
2. Insufficient demonstration of safety of Bt11 corn or
human and animal consume, and effect on the environment of
Brazil;
3. The social and cultural importance of corn in Brazil and
negative consequences of the release of transgenic
varieties over these dimensions of the Brazilian Rural
development, going against the Brazilian legislation of
protection of intellectual property of traditional
communities and indigenous people;
4. The observation of the Precaution Principle, in
accordance with Law 11.105, for the certainty that the
release of the transgenic varieties of corn will cause
direct impact in traditional, local and creoles varieties
of corn, important component of the Brazilian biodiversity,
harming, then, the environment.

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