THE CURIOUS CASE OF THE “AFES DOCUMENT” ON GM MUSTARD
- The Assessment of Food/Feed and Environmental Safety (AFES) document on GM mustard was created by the Biosafety Support Unit of Dept of Bio-Technology. DBT is one of the main funders of this GM mustard project. The Sub-Committee might have created only a brief 5.5 pages document, if at all. It is unclear if the AFES document was signed off by GEAC before being put out for public comments for 30 days. It is also unclear if this AFES document is legally valid in the first instance.
- The biosafety appraisal of GM mustard clearly did not have various areas of expertise required for such an appraisal (even within the limited numbers of safety parameters tested for).
- The study protocols were designed by the crop developers and a majority of the studies were done by themselves. One of the studies passed off as having been done by National Institute of Nutrition has been outsourced to a private laboratory.
- The progress of GM mustard this far has been in the absence of any regulatory guidelines for environmental risk assessment in the Indian regulatory regime! ERA 2016 was created only now.
- Where the sub-committee could have prescribed several tests to be undertaken by the crop developers (the need for such studies is expressed in their meeting minutes), they push various such studies to post-release monitoring of GM mustard. We present a list of such studies.
- Many illustrations are shared that show GEAC in other cases going into details of various protocols to be adopted and additional studies to be taken up. They chose not to do this with GM mustard.
- The GEAC meeting minutes show that there are inconclusive discussions and processes initiated within the regulatory body on whether HT crop applications should be considered for commercial cultivation at all, whether toxicity test conclusions being drawn are right, and whether GM mustard should undergo feeding trials or not. The regulators show no consistency in following up on their own discussions on these matters.
- Most importantly, the AFES document’s lack of reliability is exposed. While individual test reports have concluded something, the AFES concludes something else. We show with illustrations how data from weediness studies are inconclusive in the first instance, how certain claims of some tests done by the AFES document are unfounded, how and why pollen flow study conclusions are unreliable, and how agronomic evaluation is deceptive and doctored.
- Concealed data so far on the health safety front shows significant differences in compositional analyses between GM and non-GM counterparts. These are being brushed aside and attributed to agro-climatic changes which should not have been allowed firstly in a rigorous experiment. Based on falsely-established-equivalence, several other tests are being denied on GM mustard.
- Sub-Chronic Toxicity Studies reveal statistically significant differences in biochemistry parameters, histo-pathological differences and body weight gains. However, these are being brushed aside too.
WE SHOW ONCE AGAIN HOW THE AFES DOCUMENT IS UNSCIENTIFIC, UNRELIABLE, DECEPTIVE, FALSE IN SEVERAL ASSERTIONS AND INVALID IN ITS PROCESSES AND CONCLUSIONS.
THE CURIOUS CASE OF THE “AFES DOCUMENT” ON GM MUSTARD
On September 5th 2016, the Genetic Engineering Appraisal Committee (GEAC) put out a document on the website of the Ministry for Environment, Forests & Climate Change (MoEFCC) called the AFES (Assessment of Food and Environmental Safety) for environmental release of Genetically Engineered Mustard (Brassica juncea) hybrid DMH-11 and use of parental events (Varuna bn3.6 and EH2 modbs2.99) for development of new generation hybrids. While doing so, they asked for public comments on the AFES document within 30 days’ time and stated that if somebody wants to review the biosafety data, they can come all the way to Delhi and get a “dekho”. Not to be photocopied or photographed, but for any concerned citizen or expert to sit in the Ministry’s office, guarded by a designated person from the department, to do any memorizing and mental analyses to enrich the comments to be given. Provided you believed that the GEAC is serious about taking any feedback on board in the first instance. They went out of their way to stick to their modus operandi and provided no extension to the timelines nor published the biosafety data despite many senior scientists and eminent citizens writing to them.
Several of us refused to engage with their farcical process and this document is an attempt to show why it is futile to engage with a deceitful and unscientific regulator who is under pressure to clear this GMO whatever the compromise to scientific integrity in the process.
Things have come to such a pass that in a Rejoinder to an interlocutory application in an ongoing PIL on GMOs in the Supreme Court (WP260/2005), the Government has no qualms in saying that the AFES report posted by GEAC is adequate to meet the public’s interest and need for transparency. Additional detail will not provide any greater value to the public…..The petitioners want to engage in an independent review of the dossier. In no functional regulatory system anywhere in the world such independent review is entertained. The claim for independent review by the petitioners is misleading for general public by questioning credibility of Indian scientific community and demeaning of the regulatory system”. It is obvious that they don’t want the world to know how inefficient and unscientific they are.
What the government does not know or does not want to acknowledge is that it is also the scientific community in India which has been engaging with the issue of GM mustard, which is indeed questioning the credibility of the regulatory system. Numerous experts and senior scientists from different fields, from within the NARS system who do not want to come out into the open but who have many valid questions to raise, have been supplying people like me with pointers about inaccuracies and inadequacies of GM mustard safety testing and assessment. It is upto the regulators whether they will interpret it as demeaning the regulatory system, or will be grateful for the support from independent scrutiny of data, which will help the regulators fulfill their obligation in a robust fashion. The current state of affairs is also a reflection on a scientific establishment in India where dissenting views and evidence cannot be presented where the government has decided in an unscientific fashion that GM mustard is needed and safe. As inside reports reveal of a recent GEAC meeting with state government representatives ostensibly to discuss future research and field trials of transgenic crops in the country, officials of government departments are not averse to arm-twisting state governments with threats of finance cuts if everyone does not fall in line with the blind promotion of transgenics in the name of ‘biotechnology development’. It is time that the regulatory biases are exposed fully.
WHAT IS THIS AFES DOCUMENT, ITS SANCTITY AND VALIDITY? WHO WROTE IT?
The GEAC, on its website, claims that this AFES document is the “Safety Assessment Report of the Sub Committee on GE Mustard”. It says that the “sub committee has examined the dossier on food safety, environmental safety, compliance etc. and has prepared a document “Assessment of Food and Environmental Safety”.
However, we know that the Sub-Committee has probably only created a 5-page report from its assessment, and not the AFES document as put out on the regulators’ website. Information available from GEAC’s 128th meeting minutes (Agenda 4.1.1.), along with an RTI application through which two reports of the only two meetings of the Sub-Committee were obtained (On February 2nd when they finished their main work hastily within 15 days from the time of creation of the Sub-Committee, and on April 11th 2016 when they wrapped up their work) show that it is the Biosafety Support Unit of DBT which wrote this AFES document.
In the normal course of such a (sub-)committee functioning in any regulatory system, there would be a mention of the Terms of Reference, the members of the committee, their meetings and their attendance, the methodology adopted and even signatures of the members, or at least the Chair. In the normal course, such a sub-committee would have first obtained public feedback after putting out all biosafety data and then come up with its own conclusions and recommendations. GEAC’s own earlier sub-committee/expert committee reports are an illustration in case. This so-called sub-committee report-cum-RARM (Risk Assessment Risk Management) document-cum-AFES document was not your normal appraisal and review document with recommendations, however. It was not written by the sub-committee to begin with, but the BSU.
The BSU is supposed to have 16 multi-disciplinary scientists, with a couple of years’ of experience on an average each. They apparently are the ones who will suggest which guidelines should be followed, and which can be dispensed with, on a case by case basis!! They were reportedly briefed by American officials, and an Australian team trained them in document writing for risk assessment, we understand.
In the first meeting of the Sub-Committee, it is seen that the Committee recommends that the GEAC direct Delhi University to prepare the Risk Assessment and Risk Management document for consideration of the GEAC. In the second meeting of the Sub-Committee, on 11/4/2016, they decide to call the RARM document authored by the BSU as AFES, upon the recommendation from Dr S R Rao of DBT. In the end, both the names suit the purpose of Rao and others – to call something as an assessment document, and also name it as a document that has figured out future risk management too, with a pre-concluded assumption that approval will take place, and post-release risk management has to be brought into the picture!
The AFES document itself is a shoddy document which selectively puts out some bits and pieces of information, where convenient, in a manner that is opportunistic and not scientific and is mostly a weak defence document for the regulators’ lack of rigour. It is clear that those sections which have been excessively elaborated upon are those where some critical sub-committee comments have been received, and are being responded to in a pre-emptive fashion. Some such portions are also in anticipation of criticism from civil society groups too, given that some such detailed analysis was shared by a delegation on July 18th 2016.
In the end, it’s clear that the sub-committee’s main feedback came in a rushed manner within 15 days, followed by going back and forth through the BSU with the crop developer team and finally coming up with a 5.5 page report and may be not even with a signing-off the AFES document that was put out on the website. It is unclear why the GEAC did not discuss the AFES document in its full meeting before putting it out on the website – was it pressure from the PMO?
We had already pointed out to how GM mustard’s appraisal and public consultation processes are vastly different from Bt brinjal’s. It is only such scrutiny that showed that Bt brinjal cannot be claimed to be safe. It is clear that the regulators did not want to take such a risk this time. Hiding the biosafety dossier and putting out a shady document called AFES suited their purpose immensely. Meanwhile, it is insulting for any serious scientist to be reacting to the limited information available in the AFES document for any meaningful or intelligent feedback to be provided.
Some questions will not go away. What after all is the sanctity of this AFES document, even by the rule book? Is GEAC allowed to have sub-committees and should such documents be signed off by them? Should it have been put out for public feedback at all?
EXPERTISE NEEDED FOR APPRAISAL AND WHAT THE SUB-COMMITTEE HAD WITH IT
The collective arrogance of the sub-committee and GEAC, in not welcoming any independent scrutiny of the biosafety data and their arguments in the Supreme Court right now is worth going into briefly.
In the environmental safety studies, it is clear that soil scientists, plant pathologists, entomologists and within that, specialists on bees and other pollinators, mustard breeders, agronomists etc., should have been present. That is the only way that weediness and aggressiveness potential, agronomic evaluation, crossability and pollen flow potential, impact on soil microbiota, pests, diseases and beneficial organisms could have been appraised soundly by the Sub-Committee in a scenario when the rest of the GEAC is not either consulted, or has abdicated its duties of appraisal.
On the food safety front, genetic toxicologists, clinical toxicologists and pathologists to interpret the study findings properly, public health experts and nutrition scientists were important to have for reliable appraisal of safety. On the molecular studies’ front, there was a need for a molecular biologist/plant biotechnologist/geneticist.
Risk assessment does not end there. There are of course other risks related to modern biotechnology, and this GM mustard too. Therefore, there is a need for trade experts, IPR experts, sociologists, rural livelihoods experts etc.
|Expertise that was needed||Expertise that was available in the Sub-Committee|
|1. Plant Biotechnology/ Genetics/ Molecular Biology
2. Mustard breeding
3. Plant pathology
4. Soil science
7. Bee expertise (given India’s case by case approach in regulation, GM mustard appraisal should have drawn in a bee and honey quality expert)
8. Genetic Toxicology
9. Clinical pathology
11. Nutrition Science
12. Trade expertise
13. Social sciences/agrarian studies
14. Livelihoods expertise
15. Any other required area of expertise
|1. Botany/Molecular Biology/Biochemistry*
2. Plant pathology
3. Ecology/Environmental Studies
4. Plant Genetics and breeding*
(in fact two experts each as indicated by the * above had similar academic training/qualifications)
When a sub-committee with a limited combination of expertise appraises this GM mustard in a hasty fashion and when several of them also hold conflict of interest, would there be reliable risk assessment? Does their certification of safety of GM mustard actually indicate any safety at all?
SUB-COMMITTEE PUSHES STUDIES THAT OUGHT TO BE DONE NOW, TO POST-RELEASE MONITORING!
One of the most striking aspects of the sub-committee’s assessment is that they obviously find the testing inadequate, think that sticking to only the guidelines laid down is “untenable”, and then recommend several studies to be done in the post-release monitoring phase. Now, how is this justified? What is the tearing hurry to approve GM mustard now and postpone these studies? What benefits are being expected and what risks are being taken in this shortcut adopted by the sub-committee? Will planting of GM mustard in Rabi 2016 make such an impact on India’s oilseed scenario, if that is the unfounded belief? What justifies the haste? Here are some illustrations:
- “The Committee advised for continuous monitoring and further investigation for fitness and transfer of transgenic trait from DMH-11 to their progenies and feral populations that will be essential for implementing management strategies to minimize persistence and dissemination from release site…. The Committee advised that a suitable Post Release management strategy should be implemented”. (Point iv of Sub Committee’s February report)
- “Can we address the question of whether grazing of transgenic mustard by farm animals would affect them? ‘DU Reply: Only transgenic hybrid DMH-11 will be grown extensively. Barnase expresses only in the tapetum. Barstar expresses in the tapetum but also at very low levels in the leaves. However, both Barstar and Barnase have been found to be safe in sub-chronic and acute toxicity studies on model animals. Seed meal derived from GM canola lines is being fed to farm animals for more than 13 years without any ill effect’. The (sub-)Committee reviewed the reply and noted that the above answer may be taken as a post-release monitoring component” (Point 4. Of Prof K Veluthambi’s questions, Page 7 of Sub Committee’s February report). THIS OF COURSE IGNORES THE FACT THAT THE REPLY IS UNSCIENTIFIC, IGNORES HERBICIDE USE ON GM MUSTARD, IGNORES BAR GENE EXPRESSION, IGNORES THAT GM CANOLA REFERENCE IS TO A DIFFERENT SPECIES ETC.!
- “Honey made from barnase expressing Varuna and DMH-11 needs to be tested for barnase levels and biosafety. ‘DU Reply: As nectaries are well developed in barnase lines, we do not expect expression of the barnase gene. This can be checked post-release to allay any concerns’. The Committee requested that honey be collected from the vicinity of cultivated area and check whether honey accumulates barnase, barstar and bar gene proteins. This is suggested as a post release management requirement”. (Point No. 17. Dr S K Apte’s questions, Page 13 of Sub Committee’s February report). IT IS MYSTERIOUS WHY THIS STUDY COULD NOT HAVE BEEN PRESCRIBED RIGHT NOW. IT IS IMPORTANT THAT HERBICIDE CONTAMINATION IS BEING IGNORED AGAIN.
- “Prof C R Babu recommended to the Committee that long term Post release monitoring should be implemented to study (i) weediness; (ii) pollen flow to wild relatives; (iii) the impact on beneficial insects and (iv) the impact on beneficial soil microbes if any”. (Point e, Page 15 of Sub Committee’s February 2016 report). NOT CLEAR HOW THE SUB COMMITTEE WAS HAPPY WITH ONE SEASON, ONE LOCATION STUDIES IN THE FIRST INSTANCE, AS IN THE CASE OF POLLEN FLOW STUDY!
- “Effect on beneficial soil micro-organisms (as per 1989 guidelines of RCGM document) may be evaluated as a part of Post Release Monitoring”. (Sub Committee’s 2nd Meeting on April 11th 2016, Page 2). THE COMMITTEE NOTES THE SERIOUS LIMITATIONS OF THE STUDY ON SOIL IMPACTS BUT DOES NOT GO THE ADDITIONAL REQUIRED STEP OF ACTUALLY PRESCRIBING SOUND TESTING!
- “As part of sustainable use of de-regulated GE mustard in future, it is important to demonstrate that honey derived from GE mustard be tested for the absence of barnase for a definite period as a part of Post Release Monitoring”. (Sub Committee’s 2nd Meeting on April 11th 2016, Page 2).
- “The Committee opined that ‘establishment of self reproducible populations of Brassica juncea in the hills’ may be taken as a scientific question which should be addressed from a research angle in the interest of long term sustainability of GM mustard technology”. (Sub Committee’s 2nd Meeting on April 11th 2016, Page 2).
Although food and environmental biosafety elaborated in this document did not reveal any measurable risk, for sustained use of technology in breeding for newer hybrids, some post-release monitoring / stewardship is suggested as a precautionary measure. These measures include: monitoring honey bee behavior particularly with respect to presence of target proteins in honey; impact on non-target organisms and intra and inter-specific interactions. Additional measures should be taken not to include any chemicals for weed control in the package of practices. (AFES document, Page 112).
It is worth noting that it is bad enough that several important safety considerations are pushed as recommended studies to be taken up as part of post-release monitoring. In the final AFES document, even these (7 points above) are not repeated faithfully, but only selectively (italics, above).
The important question here is why should the crop developer be not asked to take up these tests now? Why should this be done after release?
Who will be made responsible for any liability for these willful shortcuts and shortcomings?
WHO CREATED THE PROTOCOLS AND WHO DID THE TESTS?
As per an RTI response obtained from CGMCP, all the study protocols were designed by Delhi University (the applicants), in association with testing labs and institutions (RTI reply included as an Annexure 1 to this note). In terms of actual conducting of tests, the following seems to be the picture:
Environmental safety assessment:
- Weediness potential: Crop Applicant CGMCP, Delhi University
- Crossability and gene flow studies: Crop Applicant CGMCP, Delhi University
- Studies on soil microbial community: IMTECH, Chandigarh (a CSIR body)
- Studies on pests, diseases and beneficial organisms: Crop Applicant CGMCP, Delhi University
- Agronomic evaluation: Crop Applicant CGMCP, Delhi University*
Food safety assessment**:
- Allergenicity assessment: Food and Drug Toxicology Research Centre, National Institute of Nutrition, Hyderabad
- Toxicity assessment (acute oral toxicity, sub chronic toxicity): Food and Drug Toxicology Research Centre, National Institute of Nutrition, Hyderabad (M/s Premas Biotech Private Limited, Manesar was involved in preparation of purified proteins for some testing.
- Nutritional and compositional analysis: Outsourced to M/s QPS Bioserve India Private Limited, though claimed and signed off as study by Food and Drug Toxicology Research Centre, National Institute of Nutrition, Hyderabad
Molecular level characterization/studies:
- Expression levels of proteins: Crop Applicant CGMCP, Delhi University
- Molecular characterization: Crop Applicant CGMCP, Delhi University
*AFES claims that these were supervised by DRMR, an ICAR body. But a DRMR RTI response indicates otherwise.
** We have already brought out a report that points out that these studies are not reliable as far as we can see – it is the same person who advised CGMCP on the studies to be taken up as part of an institutional expert committee [DR B Sesikeran] who also has a formal association with a industry-funded body called ILSI, who headed NIN at the time that these studies were done. What’s more, he also was a regulator deciding on the results of studies conducted in his institute on his own advice!
6 of the above 10 aspects of safety assessment were not only designed by the crop developers themselves but were undertaken by them. 3 other studies on the health front were done by National Institute of Nutrition, including one outsourced study (GEAC in fact has discussed in one of its meetings that such outsourcing is not to be allowed). We will let the reader decide for herself/himself whether they would like to rely on the tests and results.
HAS GEAC ALWAYS LEFT EVERYTHING TO GUIDELINES?
The 2008 guidelines created in India, in the name of harmonization with international guidelines, and supported by USAID and industry lobby groups are clearly a case of lax regulatory guidelines, and are no protection from the risks of modern biotechnology. (http://igmoris.nic.in/files/Documents%20for%20IGMORIS%2014%20july/International%20Conference%20on%20GM%20foods/Proceedings%20of%20the%20Intl.%20conference%20on%20GM%20Foods.pdf, http://www.cera-gmc.org/files/cera/docs/sabp_activities/report_fs_2006_07.pdf)
GM mustard crop developers and the unknown authors of the AFES document hide behind the 2008 guidelines for safety assessment time and again. It is interesting to note that the Sub-Committee in its first meeting said that it is untenable for the crop developer to justify lack of some pertinent studies by taking recourse to the fact that such tests were not prescribed. It is also interesting to note that the Sub-Committee ends up prescribing a test to be taken up using the 1998 guidelines in its second meeting, but in a post-release setup!
117th meeting, 119th meeting, 120th meeting, 121st meeting and 122nd meeting minutes are good examples of what GEAC can discuss and decide upon, if the members want to, irrespective of what the Guidelines say. These are also useful illustrations of how GEAC abdicated its responsibilities when it came to GM mustard.
These are meetings where GEAC is seen laying down detailed study protocols – rightly so – in the case of Monsanto, Mahyco, Metahelix, Bayer et al applications for different GMOs, both for field trials and agronomic evaluation, for environmental studies and for molecular characterization.
For instance, only commercial check hybrids are insisted upon for certain crops like maize, rice and brinjal (even for event selection trials) – 117th, 119th, 121st meetings. Here, GEAC does not hide behind unscientific and silly arguments that the proof of concept is only around herbicide tolerance or insect resistance and that better hybrids can be bred later, as is being done with GM mustard.
As another example, GEAC insists on expanding pollen flow study protocols beyond a 50 meter radius to include additional lines in an additional 50 m, after the proposed pollen trap in one case (117th GEAC meeting minutes).
Agronomic practices are discussed in detail, including chemical sprays and fertilizer use.
GEAC also did not leave soil micro-biota studies to some microbial abundance studies but insisted upon a careful assessment of impact on collembola species and earthworms too, amongst other organisms. This was however not done with GM mustard. In fact, the protocol did not even cover agriculturally important species when it comes to GM mustard safety evaluation on soil microbiota, and this has been noted by the sub-committee, but meekly pushed aside to post-release monitoring studies.
GEAC in the past has changed protocols proposed from Randomised Block Design to Alpha design where appropriate, prescribed the exact amount/quantities and numbers of samples to be tested for some parameters etc.
For herbicide tolerant crops, GEAC had prescribed tests on bio-efficacy of the herbicide, residue analysis in HT crop soil, effect of leftover residues on succeeding crops, untreated controls etc. (117th meeting). Protein expression data was prescribed to be recorded at the time of each herbicide application in one case (119th meeting). Data related to soil microflora, earthworms and soil insects related to soil rhizosphere was also prescribed to be recorded during pre- and post-spray of herbicide in an instance. (119th meeting). Visual observations on herbicide treated plots for yellowing, scorching and wilting were prescribed to be recorded. Control treatments will be manually weeded in this case. Dosage of herbicide sprays, approval of CIBRC, nature and extent of bio-degradation, residue estimations etc., were all areas of additional information sought for HT crops.
Feeding studies on broiler chicken and dairy animals were also prescribed (121st meeting).
It is clear that all of this has not been done in the case of GM mustard and has been left to the crop developers fully. The GEAC has to justify why this is so and whether safety of the technology will improve all by itself just because a public-funded, public-sector GMO comes into the picture.
In the 121st meeting of the GEAC on 18/7/2014 (soon after the new government was formed at the Centre), there is a partial discussion on whether GEAC should ever be entertaining a HT crop application for commercial cultivation at all. This is obviously a pending matter even within the regulatory body (apart from this being a policy decision for the government and for judicial adjudication in the GMOs PIL), while the Committee is also rushing headlong towards GM mustard approval, which is a herbicide tolerant crop.
“2.1. Minutes (of the 120th meeting) were confirmed with the following editorial changes:
- The Committee also took note of the following observations made by Dr PM Bhargava during the 119th GEAC meeting: ‘At this meeting, we had an important and long discussion on whether or not we should entertain any applications that relate to the incorporation of herbicide-resistant gene. Many members were strongly of the opinion that such applications should not be normally entertained as the evidence against the use of herbicide resistant crops is overwhelming. We, of course, should not stop any research work in the area but we should make it clear that no permission would be granted for field trials of such crops. It was decided that all those who are against the release of herbicide-resistant crops (their names should be mentioned in the minutes) will present evidence in favour of their view at an appropriate meeting where this item would be an agenda item’”.
What is worth noting from all these minutes is the additional questions on Herbicide Tolerant crop applications and additional requirements on the applicant – for instance, whether CIBRC permission exists or not, and a copy of the same to be furnished. The bio-degradation details. Studies on the herbicide’s eco-toxicity and residue possibilities etc. In the case of GM mustard, all of this has been ignored and left unprescribed.
On the other hand, we also have conflict-of-interest-laden members like Dr Sesikeran, who took up health safety studies for applicants, explaining the study results to the rest of the GEAC on behalf of the applicants and declaring some product or the other to be safe (Agenda 4.3.1 of GEAC’s 121st meeting is a fine example of this unacceptable component of regulation)! Similarly, Dr Akshay Pradhan, a crop developer of the GM mustard in question and also a GEAC member, gets to provide explanations to the GEAC in the same meeting on GM mustard (Agenda 4.4.9, Page 14)! This is a classic illustration of why we keep talking about removing all such members from GEAC completely.
It is important to note that the Biology Document of Brassica juncea which is supposed to have guided the biosafety testing of crops like GM mustard was prepared only in 2016. The regulators have failed in their duty but want the nation to bear the consequences for their irresponsible functioning.
It is also worth noting that even the Environmental Risk Assessment guidelines were prepared and released only in 2016. Without such guidelines in place, how did GM mustard get tested in the first instance? Isn’t this the government’s falsehood in the Supreme Court way back in 2008 too, that guidelines exist and they will be uploaded on the website?
There is another matter which is worth mentioning here, pertaining to conclusions and inferences drawn from toxicity tests (acute and sub-chronic) where the reports often conclude that “some statistically significant differences were observed, but were also within the normal range and therefore, found not to be biologically significant”. In the 121st meeting of the GEAC, this was explicitly discussed.
4.3.11. Some of the Members were of the view, that the Protocols for 14-days acute oral toxicity study and 90 days sub-chronic toxicity study may need to be revisited as the number of days and sample size of 20 animals (10 male and 10 female ) may not be adequate. It was clarified that the animal numbers used in the study are accepted world-wide for pre-clinical or non-clinical evaluation of safety and as per WHO document on safety evaluation of vaccines. Principles of statistics are the same, it is just the test material which is different. It was further stated that the size of the treatment group depends on the animal model chosen, i.e., the number of animals included in studies using non-human primates would be expected to be less than in studies including rodents. For small animal models, e.g., rats and mice, it is recommended that approximately 10 animals/sex/group be studied.
4.3.12 As regards the acceptability of the statistical variations in the toxicity studies even in the absence of any biological significance, Dr Ramesh Sonti supported by three other Members were of the view that “In acute and sub-chronic toxicity studies, for parameters where there is a statistically significant difference between control and treatment, the test should be repeated for that particular parameter/s (and not for all parameters). If there is a statistically significant difference on repetition, the matter should be investigated further. The matter should be dropped if there is no difference upon repetition. This is being suggested as a matter of ‘precautionary principle’ because these are foods that will be consumed by a large number of individuals over a long period and we would like to minimize risks. Furthermore, we should go the extra mile to minimize concerns regarding GM foods, at least in the initial years of their commercialization.”
4.3.13 The Committee after a lengthy discussion, decided to constitute a sub-committee to review the toxicity data in the context of the above discussions.
It is clear that this is an inconclusive discussion within GEAC – however, the GEAC’s sub-committee steamrollered ahead, without an independent health safety expert participating, and concluded that GM mustard is safe even with statistically significant differences in various parameters as we show later on in this document.
WHAT IS THE WITHHELD-DATA SHOWING AND WHAT IS THE AFES CONCLUDING?
- Environmental Safety Studies:
- Environmental Studies: Weediness Potential
Chapter 6, Section 6.1, pg 75-80 of AFES concludes the following about weediness potential: “The weediness potential of GE mustard hybrid DMH-11 is similar to that of the varieties commonly grown in India. There is no risk of any aggressiveness or any weediness potential in the hybrid DMH-11”. Such weediness was measured by (i) seed germination; (ii) speed of seed germination; (iii) seedling vigour; (iv) small seed size; (v) long continuous seed production and (vi) pod shattering. We present more details here.
(Permission Ref. BT/BS/17/30/97-PID dated 7/2/2011, as mentioned in Appendix Table on Pg114)
|Data in the dossier
(Permission Ref. RCGM letter No. BT/BS/17/30/97-PID dated 7/2/2011)
Ref. Report: “Studies on weediness potential and aggressiveness parameters of transgenic Brassica juncea, during Rabi 2011”
“Seed germination percentage revealed that there was no difference between the GE and non-GE hybrid”
|Lab studies showed handmade non-GE hybrid to have significantly higher percentage germination over transgenic hybrid at 10 days. (Table 1 of “Studies on weediness potential and aggressiveness parameters of transgenic Brassica juncea”, during Rabi 2011).|
|Speed of seed germination:
“The number of seedlings emerged after 5, 10 and 15 days of sowing showed that speed of germination had stabilized by 10 days in both GE and non-GE hybrid”
|Table 4 of the report shows that by Day 5, DMH-11 had 84.7% germination as opposed to 70.3% in VEH2-F1, at Day 10, it was 94.7% vs. 88% and at Day 15, it was 95.3% for DMH-11 vs. 88.3% for non-transgenic hybrid. Is the conclusion drawn by AFES right?|
|Shoot and root weight:
“The data revealed significantly higher shoot and root weight in handmade non-GE hybrid as compared to the GE hybrid DMH-11 under field conditions”
|Table 2 based on laboratory based studies, shows mean shoot length as 5.64 cms of 10 day old seedlings of VEH2-F1 as opposed to 5.21 of DMH-11. Table 3 shows 12.61 cms of mean root length of 10 day old seedlings of VEH2-F1 handmade hybrid, as opposed to 12.68 cms of DMH-11. This reveals no significant difference.
When it comes to mean shoot and root weight in gms of 15 day old seedlings grown under field conditions (Table 5), VEH2-F1 has 10.49 gms fresh shoot weight, 0.852 gms dry shoot weight, 0.141 fresh root weight and 0.043 dry root weight, as opposed to DMH-11 having 7.95 gms, 0.648 gms, 0.109 gms and 0.029 gms respectively. This reveals significant difference, with higher values in handmade non-GE hybrid.
|Long continuous seed production:
“The data on seed production shows no difference among GE hybrid DMH-11 and non-GE hybrid VEH2-F1”
|No such comparison was done between DMH-11 and VEH2-F1. AFES conclusion false.
During BRLII trials, such data on long continuous seed production for not recorded for all entries. Data presentation was left as year-wise, site-wise tables with no aggregation and no conclusions can be drawn, especially given that other hybrids were not used as comparators.
|Seed size, Pod shattering:
“The seed size of two parents of the hybrid DMH-11 ie., Varuna and EH-2 are different”
|No such comparison was done with pod shattering and no quantified data was collected. No conclusion can be drawn here, and the AFES document making it appear that it has been studied is misleading.|
|Given the above, the AFES conclusion on weediness potential is unreliable|
- Environmental Safety Studies: Crossability & Pollen Flow Studies (inter-species and intra-species)
(Ref. Letter No. BT/BS/17/30/97-PID dated 15/10/2010 for crossability studies at Bawana as cited in Appendix Table on Pg.114)
|Data in the biosafety dossier & other relevant documents
(Ref. permission letter no. BT/BS/17/30/97-PID dated 15/10/2010, for crossability studies in Rabi 2010 and 2011.
|AFES document, outside the Appendix Table of Page 115 refers to only one experiment on intra-species pollen flow in the year 2010, in DU’s Research Farm in Bawana, New Delhi (page 83). However, the season/year of inter-species crossability experiment is not mentioned.||While Bt brinjal and Bt cotton went through multiple seasons of such pollen flow studies, GM mustard was put through only one season pollen flow/crossability studies, that too in one location. Is this a reliable assessment?|
|AFES document, page 83:
Chances of escape of transgenes to related Brassica species may occur only if these are present in the receiving environment.
Successful crossability requires that the populations of donors and recipients must overlap temporally and spatially, and be sufficiently close biologically.
|A Biology document of Brassica juncea is prepared by the Indian regulators in 2016 (carefully undated on the publication of course), years after the R&D and testing begins on GM mustard, ostensibly to become the baseline for safety testing (as Minister and AS messages confirm helpfully to us)!! It can only be assumed that some guidance was indeed taken from this document for safety assessment of GM mustard in the current instance. Table 6 in page 5 lists the rapeseed mustard crops grown in India. The description of rapeseed mustard collections for our gene banks also shows how many varieties are present in different states of India. It is noted that outcrossing varies from 7.6% to 22% even though it is a predominantly self pollinated crop. While extent of wind pollination was recorded upto 11-17.5%, insect pollination is an important component. It is further noted that bees may carry pollen over long distance and have been found foraging in fields more than 4 kms away from the hive, resulting in outcross seed set.
A very low frequency of natural hybridization amongst B. rapa, B. juncea and B. napus does occur if they are cultivated closely (4.1, Pg. 11). Such hybrids are partially fertile and can set a few open pollinated seeds. B. juncea shows the second highest crossability with B.napus after B.rapa.
Low introgression varied between species and existed to an extent with wild relatives.
|“Hence, in the absence of synchronous flowering under growing season, the possibility of outcrossing with these (4) species is unlikely”.
“Seeds were harvested from the related species, germinated and sprayed with herbicide Basta. The study showed no Basta resistant plant in F1 generation – hence, it could be concluded that the likelihood of crossability of GE DMH-11 with other Brassica species under natural conditions is highly unlikely” (Pg. 85 of AFES document).
|Data of 1000 seeds planted in 2011-12 from the harvested seeds of 2010-11 season from the experiment does show that all the plans of other species are susceptible to Basta and were destroyed (Table 3).
Our comment: Should the limitation of the experimental protocol, of planting all species at the same time, rather than synchronise flowering stage for different species, become the basis for inferring that possibility of outcrossing itself is unlikely?
Also, should this study have been done only in one location in one season for reliable conclusions to be drawn and to estimate the risk potential from inter-species crossability, given the literature evidence on inter-species crossability in northern hills?
|Intra-species Crossability or Pollen Flow:
|Study limitation cannot be the basis for concluding something incorrectly. The outer plot of non-GM mustard of Pusa bold was sown only upto a distance of 50m. Further, while an inner plot of GM crop with the outer plot being non-GE mustard presents estimates of pollen flow of one kind, in reality, non-GM plots could be sown in a long contiguous fashion next to GM mustard crop, depending on plot sizes and shapes.
The relevant report in the dossier shows pollen flow upto 20 mts. Further, 7 or 9 plants out of 1000 are seen to be herbicide tolerant. This is a 0.7% to 0.9% contamination. However, nothing has been reported about bee abundance or activity here, nor the chemical treatment especially related to herbicides, if any, used in the experiment. This level of outcrossing is too low compared to the levels reported in literature.
The study being conducted in one location, in a small plot size, that too in only one season cannot be taken to be conclusive evidence on negligible outcrossing. The sub-committee of GEAC has to explain where they draw their confidence on their conclusions from.
Even while citing from the biology document of B juncea, the rate of outcrossing is being mentioned as 11 to 17.5% on page 86 of the AFES document, whereas the biology document mentions upto 22%. Literature also exists showing higher outcrossing than this, in addition to GEAC’s own assessment in the past with another GM mustard.
The Sub-Committee’s assessment in concluding that the crossing with B juncea will be similar with other non-GE hybrids is an incorrect statement since no transgenes spread from non-GE CMS based hybrids, to begin with.
Importantly, the conclusion that intra-specific crosses between DMH-11 and other varieties of B juncea would not have any selective advantage in the absence of Basta herbicide spray has no basis for that presumption (that there will be no Basta spray). There will indeed be herbicide usage by farmers.
In the box at the bottom of Page 86 it is stated, “progeny of such crosses will not have any survival advantage unless sprayed with a specific herbicide Basta”, and we contend strongly that this survival advantage does exist, and without accounting for this, any conclusion by the Sub Committee is invalid. The figure 6.2 is rendered invalid too, on these grounds.
Elaborate presentations have already been made to the GEAC on the impacts of such outcrossing and how farmers will lose out in the end, especially organic farmers.
|The Sub-Committee’s conclusion of negligible risk is unscientific, unreliable and incorrect. The outcrossing figure presented is unreliable (see the box below for another GEAC discussion on the subject). The conclusion drawn about negligible risk is unscientific given that farmers will spray herbicide glufosinate conferring selection advantage.|
It is worth noting that in 2002, Pro-Agro (a Bayer subsidiary) which walked up to the GEAC for commercial cultivation permission of a similar bar-barnase-barstar based GM mustard, reported “transgene escape” to a maximum distance of 35 meters, with an estimated 0.01% transfer at that distance. It had no information on level of contamination at distances of 1 to 5 meters. The GEAC records in its 34th meeting on 7th November 2002 that “considering the agro climatic conditions and small land holdings of Indian farmers, the Committee was of the view that the non-GM mustard seed from the adjoining fields is likely to get contaminated by male sterility barnase gene, barstar, neomycin and bar genes. This factor may affect the stability of the properties of the non-transgenic varieties. Trial studies conducted by the company also indicate the presence of male sterile plants in the progeny population of non-transgenic Brassica growing in the vicinity of transgenics. Further, in the 36th meeting of GEAC on 25th April 2003, under Agenda 3.0 A., the commercial release of transgenic mustard by M/s Proago Seeds Pvt Ltd is discussed again. Here, the following is recorded: ‘ICAR has conducted trials at only 4 locations which is not adequate. The ICAR trials indicate pollen flow upto 75 meters whereas the company has reported pollen flow upto 50 meters’”. (Relevant minutes can be accessed here)
- Environmental Studies: Impact on pests, diseases and beneficial organisms
There was detailed feedback provided in a special meeting of the GEAC on July 18th 2016 how the protocols for various studies are not rigorous enough, and how the data is unbelievable and appears cooked up. Those comments are unaddressed, and remain pertinent to the AFES section “6.4 on Studies on pests, diseases and beneficial organisms”, from page 94.
|Trial||Location||Mustard Aphid||Painted Bug||Leaf
|Cabbage Butterfly||Mustard Sawfly||Termites|
|Kumher||P||All Nil||All Nil||All Nil||All Nil||All Nil|
|Alwar||P||P||All Nil||All Nil||All Nil||All Nil|
|SG nagar||P||All Nil||All Nil||All Nil||All Nil||All Nil|
2nd Year (2011-12)
|Kumher||P||All Nil||All Nil||All Nil||All Nil||All Nil|
|Alwar||P||All Nil||All Nil||All Nil||All Nil||All Nil|
|SG nagar||All Nil||All Nil||All Nil||All Nil||All Nil||All Nil|
The above table, presented to GEAC on July 18th 2016, is an illustration of the unbelievable data on insect pests generated from the field trials.
More specifically about beneficial organisms, and within that, pollinators like bees, the following are some specific comments.
- What was essentially a rudimentary testing on bee abundance was concluded and stated to be a bee foraging study. It is not.
- Within the bee abundance study, while eye observation is one methodology, given that there are serious doubts about the competence of the people who recorded the observations and conducted the study, this methodology is not foolproof and should have been supplemented by other superior tools like videography, sweep net, soap bowl etc.
- The competence of the data enumerators during the trials is certainly under question given that the observations appear spurious and unbelievable. It is also a fact that none of the study locations have bee experts and the one location (PAU, Ludhiana) which does have a competent expert did not involve this expert in the biosafety testing.
- The data appears to be bogus from the wide variation seen in the observations and even lack of consistency in findings. No inference can be drawn from such findings. The least that should have been done is to repeat the studies to further understand the observations captured. There appears to be a serious flaw both in terms of sample locations chosen and competence of data collectors. Together, they put a question mark on the reliability of even the limited testing that was done with weak protocols.
- There is also a major contradiction in concluding that Mustard is a predominantly self-pollinating crop and showing results of yields in different treatments which actually demonstrate high outcrossing – it is obvious from the impact on yield increases that bee-keeping results in, that cross-pollination potential is very high with pollinators’ presence and activity. It is therefore important to fully acknowledge the high possibilities of cross-pollination, and therefore, the consequent impacts of GM mustard.
- The studies are completely inadequate in that they did not treat GM mustard as a herbicide tolerant crop – once released, farmers will certainly use it as a HT crop. Today, they are not using herbicides on mustard essentially because there are no HT crops. Once this is introduced, they will rationalise that spraying a herbicide is easier and cheaper than dealing with agricultural labourers for manual de-weeding. This use of herbicide will leave its own impacts on honeybees and other pollinators. The assessment of GM mustard is completely inadequate in this context. The illegal use of HT GM cotton in India is a good illustration of the point being made.
- Apart from the methodology adopted, and the inferences drawn from unreliable observations, there is the issue of the scope of the studies undertaken related to impact on Bees as well as Honey. Needless to say, mustard is a very important crop for bees and other pollinators in general, and these pollinators are in turn critical for our food security.
Impact on Pollinators and Bees:
* It is unclear why impact on other pollinator species was not studied – these are equally important in mustard pollination too.
* The tests should have included observations on diversity of species (with competent field enumerators who can distinguish between these).
* The tests should have studied actual foraging behaviour and not just abundance. This in turn should also make a distinction between nectar and pollen foraging.
* The tests should have been taken up with standard and essential protocols adopted for such assessment in EU, as per some bee experts.
* The tests should have been done on plots sown to DMH-11 farm saved seed too (with its segregation of traits in F2 generation) or on seed saved from DMH-11 X Non-transgenic mustard crop (with its outcrossing of traits).
Honey Production & Quality:
* Apart from studying impacts on several pollinators and within that, particular species of bees, there should have been studies on honey production – this then means studies that will last for several seasons and locations and not just few locations and seasons.
* Transgenic protein and herbicide residue levels should have been measured in the honey.
* Ex-ante impact assessment on trade security issues with honey from GM mustard fields should have been prescribed and done, given that our honey consignments will be rejected in some markets and would not be cost-effective/competitive in some other markets. On this hinges the livelihoods of around 5 lakh bee keeper households in the country.
Given all the above, the conclusion of the Sub Committee is irresponsible, unscientific and invalid. Even secondary literature-wise, studies do indicate significant impacts on non-target organisms, including beneficial organisms. In fact, it is unclear why the Sub-Committee hesitates to prescribe more tests that ought to have been done. There is no justification at all for not prescribing these tests right now or getting the testing done through independent capable organisations.
- Environmental Studies: Agronomic Evaluation
We have already given very detailed comments on many things that are unscientific and fraudulent about the agronomic evaluation of GM mustard, especially given that the very basis for coming up with transgenic pollination control is supposed to be claims around yield increases. Such yield increases being proven against parental lines is laughable. The only valid way of assessment is to test GM mustard against non-GM hybrids. The 121st meeting of the GEAC clearly prescribed the following: “Collect data regarding agronomic performance of the hybrid DMH-11 in comparison to national and zonal checks”. Please note that it did not say against the parents. There is absolutely no justification for GEAC to say anything opportunistic and unscientific at this point of time.
We have also shown based on raw data accessed, that in any case, any findings even within a rigged protocol are unreliable given the lack of consistency in the data (Slide 34 and 35 of this presentation, for example).
If the sub-committee is still going ahead with its unreasonable clean chit on this front to GM mustard, it is irresponsible. The evaluation being termed as a proof of concept for heterosis and hybrid vigour as compared to the parents is laughable. From where an application claimed that its permission would lead to an increase in India’s mustard yields to the point of bringing down its edible oil import bill, for a sub-committee to reach a stage where it is only ready to say that there is “presence of hybrid vigour in the hybrid” that too against the parents, is quite an admission of the failure of this GMO to prove anything in its favour. Any proof of concept on pollination control cannot be an end in itself if distinct benefits cannot be proven – such benefits can only be tested in a comparative framework vis a vis other hybrids, both in terms of superior pollination control and thereby superior heterotic possibilities.
The famous lines of the former Environment Minister describing Bt brinjal as a solution going around looking for a problem apply equally well or better to this GM mustard. Proof of concept of pollination control is not proof of concept of heterotic superiority or even superior pollination control. This has simply not been tested for and no statements of benefit can be made in favour of GM mustard on this front. In fact, if the so called environmental release is meant to breed superior hybrids with further research, the crop applicants can very well get back to RCGM for the same, and it does not require GEAC to provide any approvals for ‘environmental release’! We also have pointed out in the past that even pollination control is not adequately or rigorously proven with varying test protocols adopted in different centres which point to male sterility breakdown.
- FOOD SAFETY STUDIES
- To begin with, the acute toxicity study with purified proteins and the two sub-chronic toxicity studies with seeds and leaves are incomplete, invalid and unreliable since they did not treat GM mustard as a herbicide tolerant crop which will also carry toxic herbicide residues and would probably have combined effects of both GE and pesticides.
- The biosafety of glufosinate has not been established in the pesticides-related regulatory regime in India. In fact, use of glufosinate in mustard crop is illegal.
- No chronic health impacts assessment has been done with this GM mustard.
- It is perplexing how the sub-committee without an independent health expert has pronounced GM mustard as being safe for human consumption.
- It is also clear that the AFES green signal has come at a time when GEAC has not concluded any of its debates on related matters – related to herbicide tolerant crops, related to toxicity studies. Sub-Committees are referred to in different meeting minutes, but there is no evidence of such committees being constituted, or having come up with their final reports after comprehensive reviews.
- Here, we present evidence on how the AFES document is presenting either outright lies or selective bits of information to camouflage the lack of safety of GM mustard.
|AFES document||Data in the biosafety dossier|
The AFES document presents Table 5.1. on total glucosinolates and Table 5.2. on fatty acid composition. It is interesting to note that DMH-11 data is kept out of the tables.
“Thus the data submitted suggest that there are no significant differences in key fatty acid component levels in seeds or leaf of GE parental lines as well as hybrid GMH-11 as compared to their respective comparator”, claims the AFES document (page 58 and 59).
Table 5.3. presents data of glucosinolate and key fatty acids with DMH-11 compared with ‘normal range in commercially cultivated varieties’ and lists Varuna barnase and EH2 barstar also!
Further, it is claimed that values in hybrid DMH-11 are comparable with commercially cultivated Varuna and check RL 1359, and the nutrient compositions of DMH-11 are within the range of non-GE varuna and zonal check. How is this comparison valid?
“Conclusions: on the basis of data analysed, the compositional differences between GE line and their conventional comparators are within the range of natural variability encountered in mustard”.
“Also, hybrid DMH-11 is very similar in its composition, to the commercially cultivated varieties in India which have a history of safe use”. (page 61, AFES document)
|Composition differences between leaves and seeds of six different cultivars was taken up with samples from 3 locations.
Report of NIN is dated 2/4/2014, numbered as Study No. 2/2012. (Last field trials would have been only in 2011-12. It is not clear if the report was released in a delayed fashion). The actual testing was undertaken at M/s QPS Bioserve India Private Limited.
The Results section (No. 7, pg 124/360 and 125/360) shows that under different parameters, there were indeed significant differences for various parameters (minerals, vitamins, secondary metabolites, amino acids etc. in either the leaf or the seed.
However, the conclusion section (8) says: “The compositional analysis includes macro, micro nutrients were substantially equivalent inspite of the significant changes which may be due to agro-climatic changes”!!
It is important to note that the entire reasoning behind not taking up feeding studies or other food safety tests was an argument that no significant changes were detected in the Compositional Analysis.
However, it is clear now, that there were significant changes, but have been discounted in an unreasonable fashion.
In any experiment, agro-climatic changes should have been controlled for, to begin with, for such compositional studies.
In fact, the only valid protocol for compositional studies is to compare GM mustard with isogenic non-GM lines. In this study too, handmade non-GM hybrid VEH-2 should have been used. However, this was not done.
Importantly, the conclusion reported in the study reports is very different from the conclusions being claimed in the AFES document.
|Acute Oral Toxicity (5.5., page 64-65):
Recombinant proteins expressed in E. coli and purified, were used in this study, and each protein was administered orally, separately, to swiss albino mice.
“The data generated showed that none of the three proteins cause mortality or any adverse effect in the test animals when administered orally”.
“It may be noted that this gene (bar) is used as a selectable marker in the experiments and does not imply that basta spray is required during cultivation of the said hybrid”.
|It is interesting to note that the Coordinator of PCT in NIN which took up the testing, till 31st August 2012 was Dr B Sesikeran.
The number of animals were 8 males and 8 females for study and control groups each (32 in all) for Bar and Barstar proteins, and only 6 males+6 females in each group (control and test protein group) for barnase, showing some inexplicable lack of standardisation. These sample sizes are also below the numbers discussed on the subject in a GEAC meeting.
Taken up in Swiss Albino Mice as oral route administration, for 14 days each, as tests of different individual recombinant proteins. Approved by RCGM in its meeting on 22/11/2011 and the permission letter was BT/BS/17/30/97-PID. A concentration of 1000 mg/kg body weight was used for this test, using purified test proteins of bar and barnase. For barstar, 1700 mg/kg was the concentration used.
In the study with barnase protein, it is seen that the control group animals (12) had a small body weight gain, whereas the barnase-protein fed animals had a small weight loss. This was seen more in the case of male mice.
This study was done separately for Recombinant Bar Protein, Barnase and Barstar. However, the proteins are bound together in DMH-11.
Further, this study ignored the fact that GM mustard is herbicide tolerant and herbicide usage will be taken up by farmers.
The AFES document citing literature on pat gene to justify bar gene safety, after recording only 87% similarity between these two genes is unscientific.
|Sub-chronic Toxicity (5.6, page 65):
“The sub chronic toxicity studies with leaf and seed material showed that there was no significant difference in body weights…urine analysis, biochemical parameters and haemotology was in the normal range and similar for animals fed with normal and GE materials. No toxicologically significant adverse effects were observed in necropsy and histopathology studies of the vital organs of the test animals”.
|The sub-chronic toxicity study was 88 days’ long, done as two studies with leaves and seeds from three transgenic mustard lines, in Sprague Dawley Rats.
The relevant RCGM approval letter is BT/BS/17/30/97-PID dated 12/1/2012. Test material was received in April 2012 for leaves and actual administration was between 11th June 2012 and 8th September 2012. The test materials were lyophilized powder of leaves of 3 transgenic lines. The study was done using 120 sprague dawley rats (60 males, 60 females), divided into six groups fed with different test materials.
The results are recorded thus:
Although there was a significant difference in glucose levels in EH-2 (non transgenic) group when compared to transgenic group of DMH-11, as the values fall within the standard normal range, these variations are of no significance. (7.4.2. on Clinical Chemistry).
Except a significant difference was observed in weights of Lungs in Varuna (NT) and EH2 (NT) groups compared to transgenic DMH-11 group, all organ weights were normal in all six groups including control group. (7.5.1. on Organ Weights, under 7.5. Necropsy and Histopathology).
Focal interstitial nephritis in kidney was seen only in transgenic group animal but the occurrence was sparse and hence not considered significant. (7.5.3. on Histopathology).
Our Comment: It is unclear if the study is interpretable as equivalent to fresh leaves’ consumption.
Importantly, the study was not conducted on GM mustard with herbicide tolerant crop protocols.
|Sub-Chronic Toxicity with Seeds
“The sub chronic toxicity studies with leaf and seed material showed that there was no significant difference in body weights…urine analysis, biochemical parameters and haemotology was in the normal range and similar for animals fed with normal and GE materials. No toxicologically significant adverse effects were observed in necropsy and histopathology studies of the vital organs of the test animals”.
|Study permission letter from RCGM (No. BT/BS/17/30/97-PID) dated 12/1/2012. Receipt of test material on 17th April 2012. Study was done between 18/6/2012 to 15/9/2012. 120 sprague dawley rats were used for the study (60 male, 60 female), randomly divided into 6 groups. Test material, of Brassica juncea seed powder, was administered at 20 mg/rat for 90 days.
The report presents its findings and conclusions in the following manner:
“There were significant changes in clinical chemistry parameters but were within normal range and hence of no significance” (There was a significant difference of TBILI level in EH-2 (NT) group with respect to transgenic group of DMH-11”.
“MCV in Varuna barnase and EH-2 barstar was found to be significantly decreased as compared to DMH-11 group. Monocyte counts in Varuna barnase and EH-2 barstar found to be significantly increased as compared to DMH-11 group”. “Although variations were observed, but as all these variations fall within a normal range for rats, these cannot be considered as significant”.
“Histopathological observations of all organs were unremarkable except lungs, liver, trachea, larynx and lymph nodes which showed changes in a few animals of each group. Few of those changes were also evidence in the control group”.
“The histopathological result was also seen to be unremarkable”.
|In all these tests, statistically significant differences were indeed recorded, even though study protocols themselves had limitations. However, all such differences were ignored and discounted in the conclusions of safety drawn.|
- Bioinformatics analysis was passed off as allergenicity study, along with pepsin digestibility and thermal stability of the 3 proteins. Further, an argument the protein expression levels of the introduced genes were low is not proof of health safety of GM mustard (entire 5.4 section of page 63). The lack of safety of GMOs is from the inherent changes created by the process of genetic modification, and in the case of GM mustard, additional risks from the herbicide tolerance trait and associated herbicide usage.
- Rationalising the existence of safety based on “no reports of any observed ill effects from more than a decade of consumption, exports and trade in Canada” is unscientific. Lack of evidence is not proof of safety. If this were so, GM mustard can be approved in India straightaway without any regulatory assessment at all! Further, citing data on B napus opportunistically is not acceptable, and data on Brassica juncea should be presented.
- Sections 5.7. (Toxicity assessment of transgene proteins for livestock and wildlife, including cattle, goats and pigs) is invalid in its arguments. It is not based on any testing. Arguments based on PAT protein not tenable. The claim that substantial equivalence has been established based on no difference between GE and their non-GE juncea lines through compositional analysis is an outright lie as the conclusion in the report in the biosafety dossier shows (as opposed to falsehoods stated in the AFES document). All bullet points on Page 68 are either outright falsehoods or scientifically untenable arguments.
- We have already pointed out how ICMR guidelines as quoted on Page 68 are not the last word for safety assessment, and GEAC itself has taken various decisions in its past meetings to prove this.
- The statement of AFES that “it may be noted that GE canola is being cultivated in several of these countries for more than 20 years” is an incorrect statement given that only 3 countries have been cultivating.
- Section 5.9 is meaningless in its rationalization since no protocols have been evolved to actually test the impact of a GMO on Ayurveda or any other system of medicine.
- Section 5.10, Page 73 ‘Potential allergenicity assessment of barnase, barstar and bar proteins’’s claim that “this analysis further confirmed lack of significant sequence similarly of barnase, barstar or bar proteins to any known allergenic protein” is a matter of concern. What does lack of significant similarity mean and how much similarity actually exists?
It is worth drawing the attention of the reader here that in the 121st meeting of the GEAC, the following has been decided upon:
“4.4.10. On the issue of the toxicology studies consensus were expressed that unlike in the west, GM Canola is used as oil where as in India mustard leaves and seeds are also consumed and therefore, toxicology data should be reviewed with great caution. The Committee decided to refer the matter to the sub-committee proposed under agenda item no 4.3.
4.4.11 In addition, it was decided to obtain the following additional information:
- What are the other hybrids of Canola approved for commercial cultivation in the West and how much of the cultivated hybrids fall under barnase and barstar genes. ….
4.4.12.The Committee further decided to obtain additional information as indicated in para 4.4.11 above and also decided to refer the matter to the sub-committee proposed for the purpose of review to the toxicology data under agenda item 4.3.13 and 4.4.10”.
From all information available, no such sub-committee for reviewing toxicology data has been constituted whereas the GEAC put out an AFES document for public comments even without an independent health expert! No data has been shared, it appears, on other hybrids in the West and how much is under barnase-barstar technology.
It is also apparent from a perusal of the discussions and debates within the GEAC on this subject, that this is a matter pending to be resolved in India’s regulatory regime (on how toxicology data is to be interpreted).
The insistence that feeding studies will be taken up only if nothing is found in compositional analyses has also been violated in the context of GM mustard. Importantly, a decision on whether feeding studies are needed or not is pending, as per the same 121st GEAC meeting minutes, given below (it is worth noting that such a feeding study at NDRI, Karnal was actually proposed and discussed in the GEAC):
“4.5.2 The Member Secretary, RCGM informed that the applicant had sought exemption from feeding studies with leaves and seeds as they are not toxicological studies but basically meant to evaluate nutritional imbalance. As compositional equivalence of edible plant parts (leaf and seeds) has been established, and no Allergenicity has been observed, additional feeding studies may not be required. Further, the same genetic system has been in use for more than 15 years in several countries around the World. The matter was considered by the RCGM in its 133rd meeting held on 22.4.2014. While the RCGM agreed to the above, the Secretariat was advised to send a brief note to the GEAC with justification on why feeding studies are not required in the present case.
4.5.3 After a brief discussion on the matter, the Committee requested that the Note forwarded by RCGM may be circulated to all members of the GEAC for consideration of the case in the next meeting. Accordingly decision on the two proposals mentioned above were deferred”.
From all available information in the public domain of all subsequent meetings of GEAC and its sub-committee, such a discussion for a decision on the deferred proposals was not taken up subsequently by the GEAC.
The following reasoning is offered by us as to why the conclusions around ‘biological irrelevance of statistically substantial differences in any health safety parameter even when values are found within normal range’ are illogical and undependable:
- If the objective of the study is to check whether consumption of a GM food leads to abnormal health parameters (biochemistry, histo-pathological, body weight related etc. etc.) and whether these values fall within normal range of values for a particular species, then it is obvious that such feeding tests can be conducted straightaway on one set of animals and results verified against the normal range without having to resort to a comparative framework in the protocol;
- It is clear that the objective is not to verify only biological relevance of results, but to see in a comparative framework if GM-fed animals have significant differences in any parameter being studied, vis-à-vis the control group. Even here, it is important to note that chronic impact parameters like endocrine disruption or carcinogenic impacts are not likely to be captured in a linear, dose-dependent way. This will also be not applicable equally to both sexes whereas GEAC’s toxicology inferences have always sought to brush off early warnings of lack of safety by insisting that such differences should be dose-dependent and applicable to box sexes.
The only scientific and rational thing to be done in all such cases is to increase the number of assessments as well as do this for a long enough time to capture chronic impacts, until a reasonably reliable conclusion can be drawn, apart from more fundamental policy decisions to be taken on the subject.
In a discussion on the subject in the Sub-Committee (Page 4 of the February 2016 meeting), DU reply argues that it is important that even if there are significant differences found, such differences should be checked to see if they are in physiological range, and then one should correlate biochemical and haemotological data with histopathological changes. From available information, it appears that differences are indeed significant on all these fronts, and this should have been taken very seriously by the regulators and not ignored. If safety is being asserted by the regulators, they should justify the same by putting out all data in the public domain and explain the basis for their clean chit rather than present false conclusions as was done in the AFES document.
- Protein Expression Studies
This was taken up during October 2011-March 2012. Some findings worth mentioning are:
BAR PROTEIN: Expression of bar protein was noted in all transgenic lines, in all the tissues tested.
- Bar protein observed in the pollen of DMH-11 transgenic lines ranged between 0.159 to 0.363 µgm bar/mg of the total protein. This obviously has implications for honey quality/contamination. It is interesting to note that the sub-committee only keeps referring to barnase protein but not bar protein whose expression is driven by a double enhancer promoter in one of the parental lines.
- In the leaves of GM mustard, bar gene expression was more than 1 µgm bar/mg of the total protein in the range observed across different days of crop growth and different locations. In fact, in Bharatpur in Rabi 2011, in EH2-barstar leaves at 60 days and 110 days, it was more than 3 µgm bar/mg of the total protein. The dosage fixed in toxicity studies however looked at the average expression levels. Given large variability, how have the health effects of this been assessed?
- The highest expression levels of bar were observed in the root tissue. It is interesting to note that this was not looked at in the context of Horizontal Gene Transfer, soil pathogens or other organisms, use of herbicide glufosinate on the GM mustard crop and resultant implications.
BARNASE AND BARSTAR GENE EXPRESSION: It is seen that in DMH-11, low levels of expression of barnase are present in the whole bud. Further, barstar expression is also noted in different parts.
The following is excerpted from a GEAC discussion in the 121st meeting:
“4.4.9 Dr Sonti informed that the Barnase Protein is fundamentally a toxic protein and used as an anti-cancer drug. It was clarified that the protein is expressed only in the tapetum and is not expressed in any other tissue as per the analysis carried out in the report. Clarifications were also sought on why very low level of expression barstar is found in other tissues in addition to anthers but not barnase even though both the genes are expressed under the same tapetum specific promoter. At the request of the Committee, Dr Pradhan clarified that in the barstar construct the selection marker bar gene is driven by 35S double enhancer promoter (a stronger promoter than the normal 35S promoter) and the barstar gene by tapetum specific TA29 promoter. In the barnase construct the bar gene is driven by normal 35S promoter and barnase gene by TA29. The barstar expression observed in tissues other than the anthers could be due to the enhancer element of 35S double enhancer promoter influencing the TA29 promoter and thus resulting in very low expression of barstar in tissues other than anthers”.
It is apparent that when the DU scientists picked up the Bayer bar-barnase-barstar technology, they chose to tweak the technology to change the barnase construct ostensibly to avoid any leaky expression of the protein but also possibly to break the IPRs of Bayer in this sphere. Here, the barnase protein expression was regulated and controlled. However, when it comes to the barstar construct, the double enhancer promoter is a giveaway for the intent to commercialise herbicide tolerance trait with very high expression levels in the male parent. This is also driving the “leaky” expression of barstar in different tissues, however and the crop developers had absolutely no intent to address this.
Allergenicity testing: The low levels of protein expression in different tissues (bar gene expression is higher of course) and some bio-informatics based analysis of lack of similarity with known allergens has been used to present the safety of GM mustard in several parts of the AFES document, ignoring the fact that the health safety concerns around GMOs flow from 3-4 factors: the individual foreign genes, the changes due to the GE process, the additional risks from the chemical usage accompanying such GMOs and any combined effects, if any.
OTHER OBSERVATIONS AND COMMENTS ON THE AFES DOCUMENT:
- General comment on the AFES document: The AFES document reveals anything about GM mustard, as selectively as it does, in only 70 pages or so, within a 133 page document. There is hardly anything that can be gleaned about the biosafety test results from these 70 pages and GEAC is being outright stubborn in not revealing more information in the name of protecting IPRs which is unreasonable as already explained in detail, in person by the Central Information Commissioner in the first hearing in case number CIC/SA/A/2015/901798 and by the government in arguing in the Supreme Court that the public does not need to know more.
- Chapter 1 Introduction: The AFES document deceptively lists out several years of work done on GM mustard – the actual biosafety testing has been condensed into very few studies, and packed into four years in all (2010-2014). Any statement that hints at studies conducted from 2003-04 or from 2008 as mentioned in Page 5 of the AFES document is outright incorrect (Chapter 1 Introduction: “Studies on the biosafety of the parental lines and the resultant hybrid DMH-11 have been carried out since 2008…..”).
- Chapter 1 Introduction: The AFES document makes no mention of the fact that CMS technology exists for male sterility and hybridization possibilities. This is also a mischievous framing of the issue. Please note that CMS is included in the List of Abbreviations on Page 119 but not anywhere in the AFES document! (Chapter 1 Introduction, Page 1: “Since B. juncea is predominantly a self pollinating crop, a pollination control mechanism is required to facilitate cross-pollination for production of hybrid seeds. To achieve this, one of the parents has to be male sterile….”) (8.2. Summary: “….therefore, a transgenic technology based hybrid seed production system has been developed”.
- Table 1.1.: Before this section, there should have been some description of who prepared this document, adopting what methodology, with what participation and attendance, over how many meetings, with what inputs etc.
- Table 1.1.: “Selection marker bar required only for hybrid seed production” – ‘required’ does not mean anything when it comes to farmers using herbicide on the crop. This is a herbicide tolerant crop, period. Any lessons to be gleaned from large scale illegal cultivation of herbicide tolerant cotton should tell us about the reality of lack of regulation at the farmer level. This is a good example of regulatory incapability on the part of GEAC and mere statements like the one in Table 1.1. do not mean anything in reality.
- Table 1.1: “Compositional Analysis conducted at Food and Drug Toxicology Research Centre of NIN”: outright lie. It had been outsourced to a private laboratory.
- Table 1.1: Clearly shows that several studies have been conducted by the crop developers themselves. It also reveals that DRMR/ICAR not in the picture.
- 2. Global status of hybrid seed production technology in Brassica napus using MS-RF system deploying genes used for B. juncea: Only 3 countries have approved transgenic canola for cultivation, even though there are numerous countries which grow rapeseed around the world as the regulators’ Biology Document of B.juncea reveals. Table 1.3. should make the sub-committee and GEAC ask a pertinent question based on the information provided: Why are most countries not opting for transgenic technology? Don’t they have scientists who can develop transgenic hybrids? How are they able to produce hybrids without the use of GM technology? What are their yields like? We give the response by presenting a graph below about the situation of rapeseed cultivation in different countries which are major producers of rapeseed, and their yields in the recent past. The data has been sourced from Food and Agriculture Organisation (FAO)’s statistics.
- Table 1.4 on Rapeseed oil exports from Canada: Can the export figures of Canada be attributed to the adoption of transgenic technology, or is it to do with overall cultivation area, surplus production, and trade rules that came into being post-WTO? Are the regulators and crop developers right in putting forward Table 1.4 to defend an argument around GM canola? If yes, what is the argument and what is the evidence backing up the same?
Meanwhile, it is worth noting that the exports to India are quite negligible. This should be noted by those who think that GM canola oil imports into India justify and become the basis on which India should and can approve commercial cultivation of GM mustard within the country. Such an unsound argument ignores the fact that GM canola imports are minuscule compared to the overall edible oil consumption in India, and approval for commercial cultivation brings with it additional health and environmental risks, in addition to risks related to GM oil consumption which have not been tested by our regulators so far.
|CONSUMPTION OF IMPORTED GM CANOLA OIL WITHIN INDIA’S OIL CONSUMPTION
(All oil figures in Million Metric Tons)
|(LATEST 5 YEARS’ PICTURE)|
|Year||Total Edible Oil Consumption||Total Edible Oil Imports||Largest Import: Palmolein Oil||GM Oil Imports||%age of GM Canola in total oil consumption||%age of imported GM oils (soy & rapeseed in total consumption|
|Soybean oil||Rapeseed/ Canola oil*|
|Source: USDA GAIN Annual Reports on Oilseeds & Products.|
|Reports IN6047, IN5049, IN4026, IN3034, IN2030|
Table 1.4: GEAC has to be clarify whether all the imports listed against India have received permission from the regulators (including DGFT and whether such oil is being labeled in the market as per the Legal Metrology Act being implemented in the country). If yes, the permission details should be revealed to the nation and proof provided for regulatory clearances given and basis for the same.
- Chapter 2: Biology of Indian Mustard – the fact that this document has been created and uploaded into the public domain only in 2016 is one more proof of serious regulatory failure in the country, given the fact that a document which is supposed to be a baseline for biosafety testing has been brought into existence years after all testing has been ostensibly completed!
- 5. Weeds, major pests and diseases: “Weeds in rape and mustard crop are reported to cause approximately 20-30 per cent yield reduction”. The regulators have to make up their mind on whether they want to project weeds as a problem in mustard crop or not, since the crop developers downplay this in their anxiety to make sure that GM mustard is not rightly understood as a herbicide tolerant crop!
- 5. Weeds, major pests and diseases: It is interesting to note that the data that emerged from field trials does not reflect the pest and disease reality summarized in this section of AFES document. This reinforces our doubt that data has been doctored in the field trial reports.
- Chapter 3 Indian Biosafety Regulatory Framework: Listing out regulatory regimes from other countries does not suffice for the fact that our growing and consumption conditions are unique in themselves and we need sui generis, suitable and rigorous regulation. Further, the institutions listed out on pages 18 and 19 have proven themselves to be incapable of fulfilling the mandate of regulation capably. SBCCs and DLCs don’t exist in reality. Further, listing 1998 guidelines does not make sense since these have not been applied in the case of GM mustard. Same applies to the SOPs for field trials of 2008, which have been violated in the case of GM mustard trials. GEAC chose not to act on complaints and reports on the same.
- Chapter 3 Indian Biosafety Regulatory Framework The ERA coming into existence only in 2016 reinforces the contention of many groups and citizens, including the SC PIL petitioners that approvals have been taking place without sound guidelines in place. In fact, in the AFES document, the 1998 guidelines have been referred to in a misleading fashion as Revised Guidelines for Research in Transgenic Plants, rather than the full nomenclature of these guidelines which is: Revised Guidelines for Research in Transgenic Plants and Guidelines for Toxicity and Allergenicity Evaluation of Transgenic Seeds, Plants and Plant Parts, 1998 (http://www.moef.gov.in/sites/default/files/geac/biosafety.html). These are only for toxicity and allergenicity evaluation and have nothing to do with environmental risk assessment. It means that GM mustard had proceeded this far in a vacuum of regulatory guidelines!
- Chapter 3 Indian Biosafety Regulatory Framework (page 20): To argue that revised documents are subjected to wide ranging consultations is ironical for a government which is refusing to take cognizance of the recommendations made by the Supreme Court Technical Expert Committee’s independent report which includes Government’s nominees.
- Figure 3.1.b Step by Step process followed by the applicant for regulatory approval: It is clear that GM mustard did not follow this process of approval when parents were swapped after 2006-07 trials. In the Supreme Court reply filed by the Government of India in IA 47 of 2016 (WP 260 of 2005), under “Reciprocal change in the parents of hybrid DMH-11”, the government fails to provide any proof for formal permission having been accorded. It states “100. It is submitted to the Honourable Court that the developers had informed the RCGM in the year 2006 that after the trials of 2006-07 for all the testing of barnase/barstar system and hybrid DMH-11, Varuna barnase line and EH-2 barstar line would be used”. This is one of those sections in the rejoinder when the government failed to produce any Annexure as proof and it is amply clear that no permission has been accorded (the claim is not about permission given but information provided by the applicant which can be cooked up any time like many other cooked up materials in this GM mustard dossier).
- 2. Step by Step process followed by the applicant: This is a misleading heading since there is no clear sequence laid down or followed – tests happen in a parallel fashion as per convenience more than anything else. Further, page 23’s bullet point on confined trials’ conditions mitigating the ‘persistence of the GE plant and its progeny in the environment’ ignores and does not reveal the fact that GM mustard trials violated these conditions and we had provided photographic proof of the same to GEAC, which chose not to act on the complaint at all.
- 2. Step by Step process followed by the applicant, Page 23: “the data is generated to compare GE lines with their non-GE counterparts for various parameters” is an outright false statement since DMH-11 did not have any non-GE counterpart or equivalent control for several parameters.
- 2. Step by Step process followed by the applicant, Page 23: “In addition, the statutory bodies can exempt or prescribe generation of additional information or data on biosafety….”. It would be useful to know from the regulators why this provision was used only to exempt and not to prescribe additional data in the case of GM mustard?
- 2. Step by Step process followed by the applicant, Page 24: CCC could not have ensured compliance as claimed in the AFES document, given the timing of monitoring visits. However, they pointed out to violations which were not taken seriously by GEAC and no action initiated.
- 2. Step by Step process followed by the applicant, Page 24: “The evaluation of food/feed and environmental safety is carried out by the RCGM” – why should this be done by RCGM in the first instance when DBT has funded the project and approved the protocols?
- 3. Step by step regulatory compliance and data generation in the case of GE mustard parental lines and hybrid DMH-11, Page 24: “Based on the BRL I studies, approval was granted by GEAC for BRL II field trials in October 2014” – there was no appraisal of any data generated in the BRL I studies given that nearly all studies were claimed to be completed in the 2 years of BRLI studies as the list of studies given in the GEAC’s 121st meeting minutes show. All food safety and environmental safety studies were claimed to have been completed by then. However, GEAC does not study the results before proceeding forward and the minutes are a clear demonstration of this lack of responsible discharge of duties. It is also apparent that the final dossier was submitted in February 2016, as per the AFES document Page 25 description. This means that no further assessment, including of the many points made by civil society representatives were ever put to the developer or addressed.
- 4. Assessment of Food/Feed and Environmental Safety (AFES) – Risk Assessment Process: Invoking relevant excerpts from the ERA 2016 guidelines is laughable given that this document was not in place when GM mustard appraisal took place!
- Figure 3.2.: GEAC has to state explicitly who are the stakeholders who were involved in the decision-making process in the figure presented for “components of risk analysis” and also show proof of how they were involved in the case of GM mustard.
- Risk Context, page 26: From a perusal of the AFES document and from knowing the way the GM mustard appraisal took place, it is clear that GEAC does not know about the Risk Context – this includes socio-economic realities of poor rural female agricultural workers, about chemical use in states like Punjab, about growing and consumption conditions in India, about the impact of environmental toxins on an already malnourished population etc. etc.
- Risk Assessment, Page 26: It is apparent that GM mustard was not evaluated as a HT crop and the entire risk assessment stands invalid just on this count.
- Figure 3.3 to describe an iterative process of risk assessment, page 30: Can GEAC demonstrate what iterative processes did they run at all, and why they did not take the opportunity of rigorous evaluation before the large scale trials or BRL II of GM mustard?
- Risk assessment process for breeding stacked events, page 30: We have already shown that parental events did not undergo all studies and the claim in the last line on this page is incorrect.
- Risk Communication: “…Indian government’s commitment to clarity, transparency and accountability for decision-making processes”. This is most ironical, these claims on transparency, clarity and accountability. Here is a classic illustration of how none of these are adhered to by the Indian government and its regulators!
- Page 31: “However, such authorization shall also be subject to all other laws…relevant at that time”. The GM regulators are complicit along with the crop developers in violating the Insecticides Act in India by allowing glufosinate use on mustard crop. It is clear that no such conforming to other laws is being practiced.
- Chapter 4, Molecular Characterisation: We have already shown how inconsistent protein expression levels data is across different reports. Does this indicate stability of the events?
- Chapter 4, Molecular Characterisation: This is where the AFES document should also have provided information on all the IPRs on all the genetic materials used in GM mustard creation – the spacers, the promoters, the terminators, the desired transgenes, the constructs, the processes as well as the products, here in India and elsewhere. Any MTA terms and conditions should also have been revealed. This is where information on Glufosinate patent held by Bayer (through its subsidiary) in India should also have been revealed which is valid till 2018.
- 2. The male sterility-fertility restorer technology: “The system has the potential of hybrid seed production to bring about a substantial increase in crop productivity…amount of yield increase depending upon the parental germplasm”. Where has this potential that makes claims on yield increases depending on parental germplasm been proven or established? Only pollination control potential has been shown, that too inadequately.
- 2. The male sterility-fertility restorer technology: Claims on successful deployment of barnase-barstar system based heterosis breeding in 3 countries does not stand scrutiny from the fact that most other higher-yielding countries have not used this technology as we have already shown.
- 2. The male sterility-fertility restorer technology: Brassica juncea being described as a predominantly self-pollinating crop demands a thorough justification for how Varuna barnase male sterile line produced such good yields in the GM mustard trials!
- Page 34: “later, the elite barnase event was transferred to Varuna genotype of B.juncea while the barstar gene construct event was used to transfer the barstar gene to EH-2 genotype via backcross breeding” – GEAC has to produce evidence of permission granted for this.
- Page 39 and 40, narrative description, Table 4.1. and Figure 4.4. on double enhancer promoter driving the bar gene: “In the barstar gene construct, the bar gene, used as a plant selectable marker, is controlled by CaMV35S double enhancer promoter. This promoter confers a 10-fold increase in the expression levels over the corresponding single enhancer counterpart”. GEAC has to explain what is the purpose of using a double enhancer promoter for bar gene expression. This alone is a dead giveaway for the herbicide tolerance trait intended to be exploited by the crop developer and there is absolutely no purpose for this double enhancer promoter, especially given that it is also causing the problem of barstar expression unnecessarily at higher ‘leaked’ levels.
- Table 4.1. should be accompanied by IPRs related information on all these genetic materials/gene components introduced.
- 4. Method of genetic transformation of B. juncea: “The combining ability studies earlier had shown that hybrids between these two lines are significantly more productive than the parental varieties and the national and/or regional checks” – Outright lie. No national and regional checks were used as required and therefore, no significant productivity increase has been shown!
- 5. Characterisation of the inserted genetic material and stability of the genetic modification: “The male sterile barnase line has been analysed over ten generations under containment/field conditions and shown to stably inherit the male sterile phenotype with no breakdown of sterility” – outright lie. Data from field trials shows that male sterility broke down given that seed setting happened in bagged male sterile branches also.
- 6. Expression of the introduced genes: “Some level of expression (of barstar gene) was also observed in the leaf, stem and the root tissues. This could be due to the presence of a strong 35S double enhancer promoter driving the expression of bar gene”. This certainly has to be explained by GEAC as to why the double enhancer promoter had to be used, especially given that barstar is expressing in other parts because of this!
- 6. Expression of the introduced genes: “In addition to anthers, low levels of barstar transcripts were also observed in other tissues of hybrid DMH-11”. What then are the implications for honey quality and contamination, for instance? GEAC should put out data on the protein levels detected.
- Table 4.2. on Expression Levels of Barnase, Barstar and Bar proteins in the GE mustard lines: Interestingly, this is the only table where data is presented in a summarized fashion, across 3 BRL season trials from field grown plants, and one season of contained net house plants. However, this does not help a reader to understand the variability found across seasons and growing locations and to cross-verify whether the highest levels were taken into account when toxicity testing dosages were fixed. Given the obfuscation of data that GEAC indulges in, it is important that the full data is shared separately for each year and location for this parameter.
- 6. Expression of the introduced genes, Page 49: While description of proteins in seed and edible oil is presented, there is no information on oil cake and impact of use of such oil cake for soil amendments. What impacts are present from the oil cake of a herbicide tolerant transgenic mustard crop, sprayed with herbicides by farmers? What about cold pressed oil and oil cake specifically from such processes? Is risk assessment complete without exploring these aspects?
- 8. Functionality of the introduced proteins for male sterility and restored fertility: “…the functionality was confirmed by the absence of seed set upon self pollination by bagging the inflorescence”. Outright lie, as field trial data shows. In fact, protocols should have been standardized and this aspect tested further, rather than making false statements.
- 10. Cloning, purification and production of pure Barstar, Barnase and Bar proteins for biosafety studies: It does not help to describe such production and use of pure proteins, given that in reality, individual proteins do not exist, but they move as bound complexes. Testing did not account for this and to that extent, the tests are invalid.
- 11. Conclusions of this Chapter are questionable on how the transgene is inherited stably with no breakdown in sterility over many generations in the male sterile line.
- Chapter 5 Food and Feed Safety Studies: In this Chapter, it is rationalized that animal feeding studies are required only if compositional analysis shows statistically significant differences, especially in comparison to values/ranges for conventional varieties available in literature or recognized databases (“weight of evidence approach”). However, such differences were ignored and discounted completely as we have shown in this note.
- Page 79, Assessment of any potential weediness of GE hybrid DMH-11 and the parental lines: “The data also shows that GE hybrid DMH-11 is not prone to pod shattering”. We challenge GEAC to produce evidence for this, since no data on this has been collected other than a simple yes or no, that too not across all trials!
- Page 79: “farmers are not required to spray Basta in the hybrid GE DMH-11 field for weed control as Basta is not a recommended herbicide in the package of practices for mustard cultivation in India” – this is either a naïve statement or an over-smart one. No farmer is waiting to use only recommended chemicals in this country!
- Page 80: The statement that there is no evidence that GE herbicide tolerant crops are more invasive than their conventional counterparts is not borne out by the feral populations detected in various locations.
- 3. Studies on soil microbial community: The tests and conclusions are invalid since GM mustard has not been tested as a herbicide tolerant crop. It is also unclear whey other species like collembolan and earthworms were not prescribed in soil impact studies. The rationalization of safety based on excessive discussion on barnase protein (Page 90) is not reliable given that bar protein gets expressed in high levels in root tissues. The unscientific justification that the proteins are already widely present in nature and their presence in GE mustard is not expected to present any new toxicity risks to soil micro-organisms is laughable given that it ignores the fact of increased proteins like bar in large quantities from cultivation of GM mustard. How is it possible that the proteins will be present at the same levels with and without GM mustard? “Overall, not a single physical or chemical stressor was introduced in soil by GE mustard” is untenable given that HT mustard will certainly add to the chemical stressors to be introduced in soil. Only bacterial count was taken up in these studies and even here, there were variations that were found but ignored.
- Page 95 ignores studies that do exist of negative impacts on bees from GM crops.
SEVERAL OTHER ISSUES RAISED BY US IN THE PAST, OF A MORE FUNDAMENTAL NATURE AROUND GM MUSTARD, REMAIN UNANSWERED TO THIS DAY.
ALL THE ABOVE ARE ILLUSTRATIONS OF HOW THE AFES DOCUMENT IS UNRELIABLE IN ITS ANALYSIS AND CONCLUSIONS.
Annexure: RTI that shows that CGMCP evolved all the test protocols!
 This note has been prepared by Kavitha Kuruganti (firstname.lastname@example.org). This note seeks to show how GMOs like GM mustard which are unneeded and unsafe, can be cleared only by compromising on scientific rigor. Utmost attention has been paid to be accurate & scientific and if there are any mistakes, request that the same be pointed out to me.