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Draft 20160310,

SCIENTIFIC POSITION ON SÉRALINIS THREE PAPERS FROM 2012 TO 2016

Prof. Em. Klaus Ammann, University of Bern [email protected]

CONTENTS

Contents ...... 1

1. Seralini’s first paper on ...... 2

1.1. Introduction ...... 2

1.2. Flawed peer review process ...... 3

2. Publication of study: Manipulation of the press during the carefully planned and concerted launch on September 18, 2012 ...... 5

2.1. Clear rejection of the Séralini study by the European Food Safety Agency, EFSA, the direct response of CRIIGEN not fact based, not convincing ...... 6

2.2. Near unanimous rejection of study methodology ...... 9

2.3. Major flaws of the Seralini STUDy No.1 on tumors in sd rats ...... 12

2.3.1. The use of Sprague-Dewey rats highly prone to tumor-development for longterm studies ...... 12

2.3.2. Flawed defence of the use of Sprague-Dewey rats by ENSSER ...... 12

2.3.3. ENSSER and Séralini admit major mistake: insufficient sample size ...... 16

2.3.4. FEEd uptake not quantified ...... 16

2.3.5. Ad libitum feeding: well documented negative effects ...... 17

2.3.6. No written study plan approved by dated signature of the study director existing ...... 18

2.3.7. Final summary, a comparison of omissions and factual statements of Séralini et al. 2012 ...... 18

3. Second Seralini study in ploS ONE, STATUS 20150729 ...... 23 2

4. The latest publication of SERALINI et al.: Dead Cows of Gloeckner ...... 29

5. Caveat about pseudo-balance of views in the GM debate ...... 34

6. Final Word About Seralini And Criigen Activities, An Analysis From Jon Entine ...... 35

7. two new summaries of the worst pseudoscience publications underpinned with statistics ...... 39

8. Bibliographies related to Seralini three major papers ...... 41

1. SERALINI’S FIRST PAPER ON GM-MAIZE AND SD RATS

1.1. INTRODUCTION

The European Food Safety Authority has released in two steps a review of the paper by Seralini et al. (Seralini Gilles-Eric et al., 20120918) published in Food and Chemical Toxicology. EFSA highlights the multiple limitations of the study, both in the experimental design and in transparently reporting the available limited data set (EFSA-Statement, 20121004). EFSA waited in vain for the rest of the withheld original data (only much later Séralini handed over all the original data , but on the other hand provided CRIIGEN with data from Monsanto studies (EFSA-News, 20121022). Later, EFSA published the final report, likewise very negative: (EFSA Statement, 20121123). The opposition is – as usual – without proper scrutiny, supporting the Séralini-Study and is involved in discrediting unwelcome opinions by claiming that those critiques of CRIIGEN papers are coming from authors and units which are not independent and too close to the seed industry, this includes indeed also EFSA. A classic example of this kind of hollow polemics is the following declaration, a so called rebuttal to the massive critique against the Séralini paper, nota bene without a single scientific argument (Le Monde, 20121114).

It is easy to dismantle those opinions on lacking independency of EFSA as polemic argument-killers by learning about the criteria of EFSA web on how they meticulously choose their internal and external experts (EFSA Independence, 20121028a). The credibility of such declarations of dependency from the industry is seriously undermined by the fact that the so called ‘independent voices’ often do not reveal their own financial resources or claim in a hypocrite way their ignorance. Recently Séralini’s CRIIGEN was directly confronted with their own biased background and friends and sponsors (Riviere-Wekstein Gil, 201212), more in a broader view about independent research in: (Olenyi Sebastian, 20120228). Clearly, CRIIGEN is, as admits (in the present debate) Séralini openly, sponsored by organizations with a vested interest in organic farming, homoeopathy and anti-GMO retailing policy, documented also by a joint publication between CRIIGEN and Seven Pharma (Gasnier, Celine et al., 2011) for more details see (Seppi Wackes et al., 2011).

There is a strange kind of moral self-licensing at work (Merritt Anna C. et al., 2010), which helps to easily divide the world into good and bad guys and thus justifying to blur ethical principles. And worse: There are signs, that beyond those kinds of moral self-licensing games the dispute ends more often in court, as it happened with Marc Fellous, who got sentenced because of defamation in the French court to a hefty payment, his statement “Séralini failed in his obligation of intellectual integrity because of his dependence 3

on Greenpeace, from which he received financial assistance for his work”, however, the judges held that the other matters complained of were part of the scientific debate (Seralini-Fellous, 20110123).

The double irony is that CRIIGEN is financed by anti-GMO organizations, but it also is clear, that Séralini and his colleagues can be seen as scientists who strictly follow their own biased convictions and, after (Purchase, 2004), some of the CRIIGEN publications might fall under the third retraction category of “game- playing authors” such as (McBride et al., 1982). But as long as the original full dataset is not revealed, judgment is not really possible. But the case is open for debate, since there are clear signs of bias in previous publications like (Mesnage et al., 2012), where in a blatant case of manipulative and questionable methodology open cultures with human kidney cells in Petri dishes are sprayed with “realistic amounts of toxins” such as Bt toxins and glyphosate, see the clearly negative comments of two toxicology specialists (Chassy, B. M. & Miller, 20120222).

But let’s start in a chronological way related to the much debated publication of Séralini and his research group (Seralini Gilles-Eric et al., 20120918): their summary is full of wrong assumptions and conclusions, as we will see below:

The health effects of a Roundup-tolerant genetically modified maize (from 11% in the diet), cultivated with or without Roundup, and Roundup alone (from 0.1 ppb in water), were studied 2 years in rats. In females, all treated groups died 2–3 times more than controls, and more rapidly. This difference was visible in 3 male groups fed GMOs. All results were hormone and sex dependent, and the pathological pro-files were comparable. Females developed large mammary tumours almost always more often than and before controls, the pituitary was the second most disabled organ; the sex hormonal balance was modified by GMO and Roundup treatments. In treated males, liver congestions and necrosis were 2.5–5.5 times higher. This pathology was confirmed by optic and transmission electron microscopy. Marked and severe kidney nephropathies were also generally 1.3–2.3 greater. Males presented 4 times more large palpable tumours than controls which occurred up to 600 days earlier. Biochemistry data confirmed very significant kidney chronic deficiencies; for all treatments and both sexes, 76% of the altered parameters were kidney related. These results can be explained by the non-linear endocrine-disrupting effects of Roundup, but also by the overexpression of the transgene in the GMO and its metabolic consequences.

1.2. FLAWED PEER REVIEW PROCESS

The Journal of Food and Chemical Toxicology exists since 133 years, and is responsible for 20’000 titles and products (Elsevier et al., 2005) and it is by the sheer number of thousands of weekly publications quite a task to keep tight control in the production process including peer review. Things can go wrong in many ways in all big publishing houses, and keeping to Elsevier the Australian incidence of 5 journals having been sponsored and assumingly steered by the pharmaceutical industry in the background popped some years ago to the surface, triggering draconic measures of Elsevier closing those journals immediately after discovery (Hansen, 2009). Peer review is a hotly debated topic with thousands of publications, in the eyes of the author the worst of all quality assurance processes – with the exception of all the others…The debate involves post-peer review, open peer review etc and will eventually lead to surprising solutions. In particular, the analysis of the publication process of the paper of Séralini reveals some interesting facts: The ultimate responsibility of the Journal with an impact factor of 3.8 is in the hands of the editor in chief (specifically responsible for vision and strategy). It is obvious, that W. Hayes was not directly involved in reviewing and processing the paper, otherwise he would have noticed that his own suggestions about rat experimentation, published in a comprehensive paper (Hayes Wallace A. et al., 2011). He explicitly advises in several positions in the paper not to use Sprague-Dewey rats for food safety experimentation, as an example by citing (Weisburger & Williams, 1981):

“Sprague–Dawley rats were used briefly by the NCI both in exploratory projects and some early carcinogenicity bioassays, but the ‘‘large size and numerous spontaneous tumors in this strain’’ were considered detrimental characteristics compared to perceived advantages with F344 rats” 4

Fact is, that it was José Domingo acting as editor for this piece (approved by him personally in Nature (Butler Declan, 20120925). He is well known for his negative view on GM crops, as everyone can see in his reviews on the subject, notoriously filtering away unwelcome items in the reference list: (Domingo, J. L., 2000, 2007; Domingo, José L. & Giné Bordonaba, 2011). Domingo even has the guts (in midst of the rebuttal storm against the Séralini paper) to get cited in Nature with a ‘courageous’ statement “that the study raised no red flags during peer review” – hard to believe. Even harder to believe is the fact that José Domingo is now the editor in chief of the Elsevier Journal of food chemical toxicology, Hayes, after a short period of being an assistant editor in chief, he is now listed as an emeritus Editor in Chief.

In future, the detection of sentiments in review articles will be easier: (Tang et al., 2009):

Tang, H., Tan, S., & Cheng, X. (2009). A survey on sentiment detection of reviews. Expert Systems with Applications, pp. 10760-10773. ://INSPEC:10645113 AND http://www.ask- force.org/web/Discourse/Tang-Survey-Sentiment-Detection-reviews-2009.pdf

“The sentiment detection of texts has been witnessed a booming interest in recent years, due to the increased availability of online reviews in digital form and the ensuing need to organize them. Till to now, there are mainly four different problems predominating in this research community, namely, subjectivity classification, word sentiment classification, document sentiment classification and opinion extraction. In fact, there are inherent relations between them. Subjectivity classification can prevent the sentiment classifier from considering irrelevant or even potentially misleading text. Document sentiment classification and opinion extraction have often involved word sentiment classification techniques. This survey discusses related issues and main approaches to these problems.” From (Tang et al., 2009)

It is also difficult to accept, that W. Hayes still stands behind the specific peer review of the Séralini paper, this in a case, where dozens of red flags have been raised and the journals long standing reputation is at stake: (Hayes Wallace A., 20121017, 20121106). Nevertheless, the door is open for future scrutiny of the paper and its peer review:

“Separately, the Editors and the Publisher, Elsevier, will be considering these criticisms. We are continually reviewing our peer review processes to ensure that the journal accepts the right papers with the appropriate levels of rigor and attention to detail. If we conclude that changes need to be made to the peer review process, that will be communicated openly to readers of the journal.”(Hayes Wallace A., 20121017)

Actions like the one of Séralini contribute to heating the debate around GMOs in Agriculture with emotional values and papers with a clear result-orientation. Scientific Journals and Scientists should play a key independent role in this scenario: when they are subject to other agendas, they renounce to their function in Society, see comment of (Sinha, 2009) in Nature Biotechnology. This paper reads in its appropriate analysis of the disputes on another questionable food safety experiment with mice (Velimirov et al., 2008), which has been retracted by the Austrian government before publication, all details in (Ammann, 20120807).

For these reasons described 4 years earlier with all political ramifications in (Sinha, 2009) some rebuttals demand from the authors to retract their paper and provide a transparent list of their funding sources, and urges the Scientific Community to improve the quality of the peer-review process in order to avoid in the future similar incidences, that only create unjustified panic and do not serve Society. Peer review will have to listen to innovative authors like (Leydesdorff & Bornmann, 2012; Peerage of Science, 2012; Pulferer, 20110716; Sense about Science, 20090908). Helpful for supporting retraction in the case of Séralini is a paper of (Purchase, 2004) which describes categories of retraction types: It’s about fraud, errors and gamesmanship, in the case of Séralini its gamesmanship as the best fitting reason. In this thoughtful paper dealing with toxicology debates, an analysis of the case of Pusztai could be helpful: 5

“The report on the adverse effects of genetic manipulation of potatoes had a massive impact on the public’s perception of GM (Case study 2). Most scientists now accept that there were severe faults in the design of the experiments reported by Ewen and Pusztai (1999).Most agree that the claims made by Pusztai, before publication, to the media that it was genetic modification per se which caused the effects were an exaggeration and not warranted by the data available to him. However, the publicity surrounding this report contributed to the fact that the majority of the British public are now against the use of GM crops in the UK. From the scientific perspective, the problems inherent in the design of good toxicology experiments are not widely acknowledged. Indeed, toxicology is often viewed as an applied science with little to offer to basic science. This example demonstrates that toxicologists have much to contribute to good experimental design when the safety of foods and chemicals are being investigated. It is relatively easy to ask the question ‘is this safe’. Much more difficult is the design of the appropriate experiment to answer that question” (Purchase, 2004).

More details about the Pusztai experiment in Case study 2, emphasizing again the decisive role of experimental design, again a parallel to Séralini 2012.

2. PUBLICATION LAUNCH OF STUDY: MANIPULATION OF THE PRESS DURING THE CAREFULLY PLANNED AND CONCERTED LAUNCH ON SEPTEMBER 18, 2012

The scandalous way the authors handled the communication about the study and its dissemination is unusual: it was clearly more focused to its positive impact on the media than to the science behind their findings. It is reported by several journalists that early access to the paper before publication was only allowed upon signature of a very peculiar non-disclosure agreement: such an agreement would have prevented the journalists from approaching third-party researchers for comment. (Koerth-Baker Maggie, 20120921; PhysOrg, 20120920; Revkin Andrew C., 20120921). According to Koerth-Baker, the BBC noted in harsh criticism:

“In a move regarded as unusual by the media, the French research group refused to provide copies of the journal paper to reporters in advance of its publication, unless they signed non-disclosure agreements. The NDAs would have prevented the journalists from approaching third-party researchers for comment”. BBC in (Koerth-Baker Maggie, 20120921).

Similar statements came from Agence France Press (AFP) and from Reuters, later, several associations of science journalists also protested openly against such manipulative launch methods: International bodies on science journalism likewise protested officially against such manipulative methods: (AJSPI & Huet Sylvestre, 20121015; EUSJA Board, 20121005).

Additionally, CRIIGEN opened a dedicated website at the same time of the release of the paper, with clearly biased and defamatory dissemination material, and ready-to-use messages. The paper was published parallel to the release of books (Seralini Gilles-Eric, 2012; Séralini Gilles Eric, 2012), simultaneously also a movie of Jean-Paul Jaud, mostly based on those findings

The purpose of this novel kind of press manipulation is quite clear: The authors of that study, and The Sustainable Food Trust behind it, deliberately tried to make sure that the first stories you read about their study didn’t tell you how bad the study was (Sustainable Food Trust, 20121130), the result was that in the first days the message on the huge rat tutors caused by GM food was spread in multiple press products in a highly populist way.

The first one is the peculiar way the authors handled the communication about the study and its dissemination: a very unusual strategy for researchers, more focused to its impact on the media than to the science behind their findings.

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2.1. CLEAR REJECTION OF THE SÉRALINI STUDY BY THE EUROPEAN FOOD SAFETY AGENCY, EFSA, THE DIRECT RESPONSE OF CRIIGEN NOT FACT BASED, NOT CONVINCING

DG Sanco officially asked EFSA on Sept. 26, 2012 to review the scientific publication and ask any clarification needed to the authors of the paper. EFSA answered the same day that they will take on the job, consulting member states and the CRIIGEN author team for additional information if required. EFSA set up a multi-disciplinary task force which will analyse the paper by Séralini et al and publish an initial scientific review as the first step in a two-stage process. (EFSA, 20120926). A week later the first review was published, with a clearly negative judgement (EFSA Statement, 20121004):

“Considering that the study as reported in the Séralini et al. (2012) publication is of inadequate design, analysis and reporting, EFSA finds that it is of insufficient scientific quality for safety assessment. Therefore EFSA, concludes that the Séralini et al. study as reported in the 2012 publication does not impact the ongoing re-evaluation of glyphosate, and does not see a need to reopen the existing safety evaluation of maize NK603 and its related stacks.” (EFSA Statement, 20121004).

The reaction to this first negative assessment followed promptly, slamming EFSA for “institutionalized pseudo-science”: (GM-Free-Cymru, 20121004; GM-Watch LePage Corinne, 20121007). Both texts are completely devoid of scientific arguments, here just one symptomtic example:

There is no point in seeking to refute the silly points made by the tame “experts” quoted by the SMC—all of whom have powerful vested interests in the GMO research and development fields (3). They, after all, will all lose their jobs and their status (not to mention their own self-esteem) if GMOs really are proved to be toxic to mammals.(GM-Free-Cymru, 20121004)

On October 22 2012, EFSA provided CRIIGEN with the additional Monsanto data requested (anybody can obtain those usually confidential data on special request, which CRIIGEN missed to acknowledge officially): (EFSA-News, 20121022).

This all did not really help to trigger a more fact based scientific debate with CRIIGEN, as the following example demonstrates (the statement taken from the CRIIGEN Frequently Asked Questions prepared in the time of the first EFSA assessment (CRIIGEN & Seralini Gilles-Eric, 2012), the details will be treated in the following chapters.

Have you compared your results with those of the Japanese study of Sakamoto or another? Contrary to what you say you’re not the first to study the safety of one GMO for 2 years. Yes. None has been as comprehensive as ours, and none is on the maize NK 603 beyond 3 months.

Comment a) The research group of Sakamoto published two long term studies (Sakamoto, Y. et al., 2008; Sakamoto, Yoshimitsu et al., 2007), both were certainly done more professional and comprehensive than the Séralini study, and most important, they were done with an appropriate F344 rat strain, much better suited for long term experiments, but shunned as “not comprehensive” because of the positive results. The summary:

A chronic feeding study to evaluate the safety of genetically modified glyphosate-tolerant soybeans (GM soybeans) was conducted using F344 DuCrj rats. The rats were fed diet containing GM soybeans or Non-GM soybeans at the concentration of 30; in basal diet. Non-GM soybeans were a closely related strain to the GM soybeans. These two diets were adjusted to an identical nutrient level. In this study, the influence of GM soybeans in rats was compared with that of the Non-GM soybeans, and furthermore, to assess the effect of soybeans themselves, the groups of rats fed GM and Non- GM soybeans were compared with a group fed commercial diet (CE-2). General conditions were observed daily and body weight and food consumption were recorded. At the termination (104 weeks), animals were subjected to hematology, serum biochemistry, and pathological examinations. There were several di#erences in animal growth, food intake, organ weights and histological findings between the rats fed the GM and/or Non-GM soybeans and the rats fed CE-2. However, body weight and food intake were similar for the rats fed the GM and Non-GM soybeans. Gross necropsy findings, hematological and serum biochemical parameters, and organ weights showed no meaningful difference between rats fed the GM and Non-GM soybeans. In pathological observation, there was neither an increase in incidence nor any specific type of 7

non-neoplastic or neoplastic lesions in the GM soybeans group in each sex. These results indicate that long-term intake of GM soybeans at the level of 30; in diet has no apparent adverse effect in rats. (Sakamoto, Y. et al., 2008)

Comment b) according to (Snell et al., 2012), there exist 24 long term, multigenerational studies, it is simply anti-GMO propaganda to make people believe that there are no long term studies existing. See a section below about the regulatory reasons why 90-day studies are in many cases not even required, only in special situations necessary. And more: the recent opinion, 2-years studies should be done (EFSA News, 20121218) are not science based, but clearly politically motivated.

The aim of this systematic review was to collect data concerning the effects of diets containing GM maize, potato, soybean, rice, or triticale on animal health. We examined 12 long-term studies (of more than 90 days, up to 2 years in duration) and 12 multigenerational studies (from 2 to 5 generations). We referenced the 90-day studies on GM feed for which long-term or multigenerational study data were available. Many parameters have been examined using biochemical analyses, histological examination of specific organs, hematology and the detection of transgenic DNA. The statistical findings and methods have been considered from each study. Results from all the 24 studies do not suggest any health hazards and, in general, there were no statistically significant differences within parameters observed. However, some small differences were observed, though these fell within the normal variation range of the considered parameter and thus had no biological or toxicological significance. Why did you choose Sprague Dawley rats? (Snell et al., 2012).

The second EFSA final statement from November 23, 2012 on Séralini’s paper did not change the thoroughly negative assessment: (EFSA Statement, 20121123), the verdict remained clear and the disappointment explicitly mentioned that EFSA was not able to see the raw original data:

“EFSA reaches similar conclusions as in its first Statement (EFSA 2012). The study as described by Séralini et al. does not allow giving weight to their results and conclusions as published. Conclusions cannot be drawn on the difference in tumour incidence between treatment groups on the basis of the design, the analysis and the results as reported. Taking into consideration Member States’ assessments and the authors’ answer to critics, EFSA finds that the study as reported by Séralini et al. is of insufficient scientific quality for safety assessments. EFSA concludes that the currently available evidence does not impact on the ongoing re-evaluation of glyphosate and does not call for the reopening of the safety evaluations of maize NK603 and its related stacks. EFSA’s evaluation of the Séralini et al. article is in keeping with its role to review relevant scientific literature for risk assessment on an ongoing basis to ensure that the advice it provides is up-to-date.” (EFSA Statement, 20121123)

And a subsequently published more detailed review of Agnes Ricroch on the genomics aspects of long term studies (Ricroch, A. s. E., 2012) is also contradicting the Séralini long term study results:

Despite the fact that a thorough, lengthy and costly evaluation of genetically engineered (GE) crop plants (including compositional analysis and toxicological tests) is imposed before marketing some European citizens remain sceptical of the safety of GE food and feed. In this context, are additional tests necessary? If so, what can we learn from them? To address these questions, we examined data from 60 recent high-throughput ‘- omics’ comparisons between GE and non-GE crop lines and 17 recent longterm animal feeding studies (longer than the classical 90-day subchronic toxicological tests), as well as 16 multigenerational studies on animals. The ‘-omics’ comparisons revealed that the genetic modification has less impact on plant gene expression and composition than that of conventional plant breeding. Moreover, environmental factors (such as field location, sampling time, or agricultural practices) have a greater impact than transgenesis. None of these ‘-omics’ profiling studies has raised new safety concerns about GE varieties; neither did the long-term and multigenerational studies on animals. Therefore, there is no need to perform such long-term studies in a case-by-case approach, unless reasonable doubt still exists after conducting a 90-day feeding test. In addition, plant compositional analysis and ‘-omics’ profiling do not indicate that toxicological tests should be mandatory. We discuss what complementary fundamental studies should be performed and how to choose the most efficient experimental design to assess risks associated with new GE traits. The possible need to update the current regulatory framework is discussed.

The answer of the Séralini research group (confronted in the same Journal Food and Chemical Toxicology with many other critical contributions) (Seralini et al., 20121107), see the table p.3/4, which does not add to clarification, since the remarks from critique on the left side of the table are not referenced, often simplified and stripped of context. And the answers are sometimes plainly wrong or contain baffling statements beyond scientific common sense, really not helpful for the credibility: just a few examples:

“No reference groups: response: “Reference groups add irrelevant variability with non-substantially equivalent diets”…..

Comment: Quite an exotic statement, probably a rarity in the printed scientific literature.

Another example: 8

“Ad libitum feeding” response: In accordance with guidelines and usual practices”

Comment: The author group at CRIIGEN seems to be ignorant of the extensive literature on tumor-inducing ad libitum feeding, see more details in a section below, here just one citation: (Gumprecht, L. et al., 2009):

„Comparative data from two-year studies of Ad Libitum (AL)-fed CD controls (1977 to 1991) versus Diet Restriction (DR)-fed controls (1993 to 2005) were consistent with cross-sectional studies of 750 AL or DR-fed CD rats started in 1995. Moderate DR feeding improves the CD rat model in toxicology/carcinogenicity studies by allowing one to better distinguish the onset of background aging lesions, spontaneous tumors, and control group mortality from true drug treatment effects.” (Gumprecht, L. et al., 2009)

Fig. 1 Diagram of hypothesis for the pathogenesis of CRD in AL-fed rats. The &dquo;+&dquo; signifies a feedback effect on remaining nephrons. See text for discussion. From (Gumprecht, L. A. et al., 1993), Fig. 4.

(Gumprecht, L. A. et al., 1993) modified the models of (Brenner, 1985; Fogo & Ichikawa, 1989) for the pathogenesis of CRD (Chronic Renal Disease), they demonstrate that glomerural hypertrophy is initiated by Ad Libitum feeding, which causes an increased glomerular hydrostatic pressure in some glomeruli.

One of the major critical points of the study is highlighted by (Hammond, Bruce et al., 20121107): The strict rule should be, that an experiment follows a previously fixed study plan, in the criticized study the plan was admittedly changed in mid-way from a normal sub-chronic study to a long term 2-years study, which would have required much higher animal numbers per group,

‘‘The study should be conducted in accordance with the study plan’’. Apparently, the authors’ original intent was not to 29 conduct a carcinogenicity study ‘‘. We had no reason to settle at 30 Q1 first for a carcinogenicity protocol using 50 rats per group.’’ 31 (Seralini et al., 2012), but at some point during the in-life phase, 32 they changed the purpose of the study by extending it for 2 years 33 to assess potential carcinogenicity. Assuming they had a protocol 34 at the start of the study, they did not follow it as they substantially 35 altered the purpose and the design of the study while it was in progress 36. This should be considered a violation of GLP guidelines as the 37 study was not conducted in accordance with the original study 38 plan. If they wanted to carry out a carcinogenicity study, they 39 should have terminated the existing study, and prepared a new 40 study plan adapted from OECD TG 453. They did recognize, as sta-41 ted above, that they needed a larger number of animals (a mini-42 mum of 50 9

rats/sex/group) for a carcinogenicity study, instead of 43 the 10 rats/sex/group that they had in their existing study. For rea-44 sons which will be discussed later, their study did not have enough 45 animals to draw any meaningful conclusions. (Hammond, Bruce et al., 20121107)

Strangely enough, the Séralini research group first admitted that the small number of rats in each comparison group was a mistake in the ENSSER statement (ENSSER Statement Seralini, 20121005), but later found excuses with fluffy arguments, see in the table:

Number of rats per group comment: OECD 408 (90-day toxicity study) 10 animals per group OECD 452 (Chronictoxicity study) 20 animals per group but at least 10 animals per group are studied for hematological and clinical biochemical function

Comment: the statement is faulty in two ways: a) for carcinogenicity studies, it is important to use OECD guideline 451 (OECD Guideline 451, 20090908), besides an important number of precise prescriptions are made, related to the number of animals recommended:

Both sexes should be used. A sufficient number of animals should be used so that a thorough biological and statistical evaluation is possible. Each dose group and concurrent control group should therefore contain at least 50 animals of each sex. Depending on the aim of the study, it may be possible to increase the statistical power of the key estimates by differentially allocating animals unequally to the various dose groups, with more than 50 animals in the low dose groups;. (OECD Guideline 451, 20090908) b) In the guidelines No. 452 on toxicology studies (OECD Guideline 452, 20090908), which accompanies No. 451 and where specific additions are made in the prescriptions for non-carcinogenic toxins, the number can be lower, minimally 20 animals per sex, per group at each dose group (makes a total of 40 rats per dose). Nowhere a minimal group of 10 rats is recommended, the number given in the right answer section of the table cited above is misleading and smells as a cheap excuse to justify the change in experimental design at half way.

“Both sexes should be used. A sufficient number of animals should be used so that at the end of the study enough animals in every group are available for thorough biological and statistical evaluation. For rodents, at least 20 animals per sex per group should normally be used at each dose level, while for non-rodents a minimum of 4 per sex per group is recommended. In studies involving mice, additional animals may be needed in each dose group to conduct all required haematological determinations.”(OECD Guideline 452, 20090908)

2.2. MULTIPLE REJECTION OF STUDY METHODOLOGY AND RESULTS

Scientists reacted bluntly and rejected the study with clear statements: (Wager Rob et al., 20120924 and 2013):

Wager Rob, Giddings Val, signed by Ammann Klaus, Bradford Kent, Burachik Moises, Cetiner Selim, Chassy Bruce M., De Souza Lucia, De Stefano- Beltran Louis, Fellous Marc, Fedoroff Nina, Herring Ron, Klaus, J., Leaver Chris, Lerayer Alda, McHughe Alan, I., M. H., Newell-McGloughlin Martina, Potrykus Ingo, CS, P., Rao Kameswara, Sabah, A., Sivramiah, S., Strauss Steven, & Trewavas Antony. (20120924 and 2013). Letter to the Editor in Chief of Food, Chemical Toxicology (Elsevier), to request a serious reconsideration of the recent paper by Seralini et al. alleging tumorigenesis in rats resulting from consumption of corn derived from crops improved through biotechnology. pp. 1-4. http://www.ask- force.org/web/Seralini/Wager-Letter-Editor-Hayes-Seralini-20120925.pdf The full text below:

“Dear Dr. Hayes:

We write to you, as Editor in Chief, to request a serious reconsideration of the recent paper by Seralini et al. alleging tumorigenesis in rats resulting from consumption of corn derived from crops improved through biotechnology (Seralini, Clair, et al., 2012). As you are undoubtedly aware, the use of molecular methods to improve crop plants, now known as GMOs, continues to be a highly controversial subject globally despite the absence of evidence, to date, of human, animal or environment harm. The paper by Seralini et al. makes claims that contradict a large body of literature on the subject, reviewed recently in your journal by (Snell Chelsea et al., 2011). This review, analyses by serious scientific bodies, including the U.S. National Academy of Sciences and the Royal Society, as well as the European Union’s recent overview of 25 years of biosafety research on GMOs, all conclude that there are no negative health impacts specifically attributable to the use of molecular methods of crop improvement. Moreover, the 10

herbicide glyphosate, which affects an enzyme present in plants, but not animals, has a short residence time in the environment and a long history of safe use, as does the bacterium Bacillus thuringiensis, from which the so-called ‘‘Bt’’ gene was transferred to a number of crops to render them resistant to certain kinds of insect pests. Seralini et al. make the extraordinary claim that rats fed GM corn, with or without added glyphosate, develop tumors earlier in life and die prematurely compared with controls, attributing enhanced morbidity and mortality to consumption of the GM corn and herbicide. Such extraordinary claims must be based on sound and extensive evidence, as they are guaranteed to cause – and indeed, have caused – widespread alarm. As detailed below, this study does not provide sound evidence to support its claims. Indeed, the flaws in the study are so obvious that the paper should never have passed review. This appears to be a case of blatant misrepresentation and misinterpretation of data to advance an anti-GMO agenda by an investigator with a clear vested interest. We find it appalling that a journal with the substantial reputation of FCT published such ‘‘junk’’ science so clearly intended to alarm and mislead. In view of the importance of the ability to use modern molecular methods of crop improvement to increase the global food and feed supply and decrease the deleterious environmental impacts of conventional agriculture, we appeal to you to subject the paper to rigorous re-review by appropriate experts and promptly retract it if it fails to meet widely held scientific standards of design and analysis, as we believe it fails to do. We make this request for you to reconsider the paper because it falls short of the customary scientific and ethical standards in several specific regards: The experimental design is flawed, using far fewer animals per treatment (10) than dictated by the OECD guidelines mentioned (but not cited) in the paper (N = 50; see http://www.oecd.org/science/biosafety-biotrack/42470554.pdf). The reader is not informed that the rats used in the study, Sprague-Dawley rats, fed ad libitum diets, would be expected to develop tumors in patterns fully consistent with what the paper reports, vitiating the authors attempt to link the observed tumors with any specific dietary components. There is an abundant literature on these rats, and their responses to ad lib/ restricted diets, which the authors cite in an incomplete and entirely misleading way. The experiment lacks appropriate controls (i.e., at least 50 individuals, fed a measured diet of confirmed identity differing from tested diets only by absence of inserted DNA; a robust experiment would also include a random, unrelated diet, e.g., one derived from organic maize). Inappropriate and non-standard statistical tests were used, rendering meaningless any interpretations of the results reported – robust statistical tests of raw data to determine whether or not differences are statistically significant must be used, not mere reporting of percentages or irrelevant and exotic tests of no value (e.g., OPLS-DA). Critical details on how much food was consumed by each rat are absent, making it impossible to establish any dose/response relationship. The identity of the ‘‘control’’ diet (i.e., ‘‘non GM’’ was not confirmed, and details on food preparation methodology were not provided. The animals were not euthanized in a timely manner to eliminate unnecessary pain and suffering, as stipulated by both European and U.S. animal research guidelines. The underlying and complete data are being withheld, not shared with other scientists, as is required by Elsevier’s published policies (‘‘Authors may be asked to provide the raw data in connection with a paper for editorial review, and should be prepared to provide public access to such data (consistent with the ALPSP-STM Statement on Data and Databases), if practicable, and should in any event be prepared to retain such data for a reasonable time after publication...’’ http://publicationethics.org/files/u2/New_Code.pdf ). Thank you in advance for your consideration, sincerely, Robert Wager Vancouver Island University Canada”

There were an unusual high number of other letters to the Editor Hayes, all published in the Food, Chemical Toxicology: (Barale-Thomas, 2013; Berry, 20121107; Cockburn, 20121107; de Souza & Macedo Oda, 2013; Dung & Ham, 20121107; Folta, 2014; Grunewald & Bury, 20121107; Hammond, B et al., 2013; Hammond, Bruce et al., 20121107; Heinemann, J., 20121107; John, 2014; Langridge, 20121107; Morandini Piero, 20120920; Ollivier, 20121107; Pilu, 20121107; Roberfroid, 2014; Rosanoff, 2014; Sanders et al., 2013; Souza, 20121107; Tester, 2013, 20121107; Trewavas, 2013; Tribe, 20121107; Viljoen, 2013; Williams, 20121107), the great majority negative, supporting letters only from well known GMO opponents like e.g. Heinemann.

Here also an unusual large number of harsh critiques published by official organizations, which are largely consistent with the EFSA statement (Academies Francaises, 20121018-19; Belgian Biosafety-Advisory, 20121019; BfR-Bund, 20120928; CNRS, 20120927; COMETS-CNRS, 20121002; Council-Biotechnology, 11

20120920; CTNBio et al., 20121024; EFB-Statement et al., 20121004; EFSA-Statement, 20121004; EFSA, 20120926, 20121009, 20121011; EuropaBio, 20120919; EUSJA Board, 20121005; FSANZ Food Standard A- NZ, 20121011; Haut Conseil Biotechnologiques, 20121019; Health Canada, 20121025; Liberation, 20121002; PRRI, 20121129; Schorsch, 20121107; Science-Media, 20120919; Scientific-Alliance, 20120921; VBio, 20120921; VIB-Report, 20121008). Instead of summarizing the major points of each position, it is possible to summarize the flaws in several sub-chapters of 2.3.

Some content is also coming from numerous publications in other journals, magazines, blogs etc: Just two of them standing out is mentioned here in extenso: (Arjó et al., 2013):

Reactions comments and rebuttals of critique of the first Séralini long term study published outside the Food Chemical Toxicology journal are numerous, a proof that the echo of the study (not speaking about the hundreds of press products) was gigantic.

(ABNE, 2012; Aris, 2012; Bales et al., 2012; CERA (AGBIOS), 2012; Chapin et al., 2013; Clair, E. et al., 2012; Clair, É. et al., 2012; Collotta et al., 2013; CRIIGEN & Seralini Gilles-Eric, 2012; de Liz Oliveira Cavalli et al., 2013; Dietrich, D. et al., 2013; Dietrich, D. R. et al., 2013; Edwards & Beetles, 2012; Evers Elsa & Greenpeace, 2012; Fang & Casadevall, 2012; Fang et al., 2012; Frazier et al., 2012; Fresco, 2013; Friston, 2012; GMSAFOOD et al., 2012; Goldstein et al., 2012; Harleman et al., 2012; Hartung et al., 2012; Heinemann, J. A. et al., 2013; Henderson Mark, 2012; Herman & Raybould, 2013; Houllier, 2012; Jany, 2013; Jany Klaus, 2013; Juranek et al., 2013; Kamle & Ali, 2013; Keogh, 2012; Kim et al., 2013; Knasmueller et al., 2012; Kouretas et al., 2013; Kuntz, 2012; Kuroiwa et al., 2013; Liu et al., 2012; Loutchanwoot et al., 2013; Mann et al., 2012; Martinelli et al., 2013; Meldolesi, 2012; Mesnage, R et al., 2013; Mesnage, R., B. Bernay, et al., 2013; Mesnage, R., E. Clair, et al., 2013; Mesnage et al., 2012; Meyer & Hilbeck, 2013; Mink et al., 2012; Nau, 2013; Nicole, 2012; Nielsen, 2013; Paganelli et al., 2012; Piersma et al., 2013; Ricroch, A. E., 2013; Robinson et al., 2013; Romano et al., 2012; Romeis et al., 2013; Seok et al., 2013; Séralini Gilles Eric, 2012; Tanaka et al., 2012; Van Eenennaam, 2013; Vandenberg et al., 2012; Vernet et al., 2012; Williams Amy Lavin et al., 2012; Woubit et al., 2012; Zobiole et al., 2011)

Actions like Séralini’s, contribute to heating the debate around GMOs in Agriculture with emotional values. Scientific Journals and Scientists should play a key independent role in this scenario: when they are subject to other agendas, they renounce to their function in Society, see comment of (Sinha, 2009) in Nature Biotechnology.

For these reason some rebuttals demand from the authors to retract their paper and provide a transparent list of their funding sources, and urges the Scientific Community to improve the quality of the peer-review process in order to avoid in the future similar results, that only create unjustified panic and do not serve Society. (Leydesdorff & Bornmann, 2012; Peerage of Science, 2012; Pulferer, 20110716; Sense about Science, 20090908)

See retraction chapter below

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2.3. MAJOR FLAWS OF THE SERALINI STUDY NO.1 ON TUMORS IN SD RATS

2.3.1. THE USE OF SPRAGUE-DEWEY RATS HIGHLY PRONE TO TUMOR-DEVELOPMENT FOR LONGTERM STUDIES

One of the major flaws (mentioned by all critiques without exception, excellent example (Miller Henry I. & Chassy Bruce M., 20120925)) is the use of the wrong rat strains (Sprague-Dewey): there is a rich literature based on hard experimental long term data which demonstrates, that rats used by Séralini et al. are definitely not suited for their long term experiments of two years: they reach in this period their natural life span, and in particular the Sprague-Dewey rats are highly prone to get tumours of many kinds within months. The control rats develop towards the end of the two years experimental period between up to 70 and 85% spontaneous tumours, lesions and neoplasms, although kept in healthy conditions and no test toxins administered, here only a small sample of publications: (Anonymous-Rat-List, 2003; Dinse et al., 2010; Durbin et al., 1966; Kaspareit & Rittinghausen, 1999; Krzyżowska, 2010; McMartin et al., 1992; Pettersen et al., 1996; Planas-Silva et al., 2008; Poteracki & Walsh, 1998; Prejean et al., 1973; Thompson, Samuel W. & Hunt, 1963; Thompson, S. W. et al., 1961; Wahle et al., 1999; Walsh & Poteracki, 1994; Welsch et al., 1970).

Séralini’s defence in (ENSSER Statement Seralini, 20121005) is astonishingly weak, since he and his colleagues cite 12 publications (Conti et al., 1988, online 2006; Flamm et al., 2003; Guiterrez et al., 2011; Hack et al., 1995; Klimisch, Deckardt, Gembardt, Hildebrand, Kœttler, et al., 1997; Klimisch, Deckardt, Gembardt, Hildebrand, Kuttler, et al., 1997; Lee et al., 2010; Maltoni et al., 1988; Minardi et al., 2002; Morcos & Camilo, 2001; Perri et al., 1981; Soffritti et al., 2006; Voss et al., 2005) with SD rat experiments, seemingly justifying thus the use of SD rats in his own experiments. All those papers are critical towards the use of SD rats, as can be seen from the citations of the downloaded full texts, detailed comment in the subsequent sections

Séralini and his crew also seem to be ignorant (or do filter away) of unwelcome other publications dealing in a critical way with results of rat experiments, and often the target are the SD rats not suited for any long term experiments, despite its frequent use in former years – and defended with an obvious commercial bias by the producers of those rats.

An early paper of McMartin et al. demonstrated that Sprague-Dewey control rats in 2-year studies were highly prone to spontaneous varieties of cancer (McMartin et al., 1992)

Historical data are presented for neoplasms and related proliferative lesions from 1,170 Sprague-Dawley rats that served as controls in 9 carcinogenicity ( 2 year) studies conducted in the Safety Evaluation Facility of Ciba-Geigy Corporation, Summit, New Jersey. The most common neoplasm was pituitary adenoma, which occurred in 62.2% of the male and 84.7% of the female rats. Incidences of other neoplasms that occurred in more than 6.0% of the rats were, for males, benign pheochromocytoma (19.0%), cutaneous keratoacanthoma (7.9%), pancreatic islet cell adenoma (7.5%), benign testicular interstitial cell tumor (6.50/0), and thyroid C-cell adenoma (6.5%). For females these incidences were mammary fibro-adenoma (3 I .3%), mammary adenocarcinoma (1 6.8%), and mammary adenoma (6.5%). Focal cortical hypertrophy/cystic de- generation of the adrenal, a focal non-neoplastic lesion of zona fasiculata cells that often degenerate into large cysts, was present in 23.4% of all male and 82.7% of all female rats. Criteria for the differential diagnoses of selected neoplasms and related lesions are presented.

2.3.2. FLAWED DEFENCE OF THE USE OF SPRAGUE-DEWEY RATS BY ENSSER 13

The Séralini publication has provoked (compared with the poor quality of the paper) a storm in press and activist websites, including a deeply flawed rebuttal of ENSSER: (ENSSER Statement Seralini, 20121005)

ENSSER Statement Seralini (20121005), Questionable Biosafety of GMOs, Double Standards and, once again, a "Shooting-the-Messenger" style Debate, ENSSER, European Network of Scientists for Social and Environmental Responsibility, No. pp. 8, ENSSER Statement on Séralini et al. (2012) publication and reactions evoked Hyderabad, India at COP-6 Meeting, http://xa.yimg.com/kq/groups/18208928/312025102/name/ENSSER_Seralini.pdf AND http://www.ask-force.org/web/Seralini/ENSSER-Statement- Seralini-20121005.pdf

Summary of ENSSER:

The European Network of Scientists for Social and Environmental Responsibility (ENSSER) welcomes the study “Long term toxicity of a Roundup herbicide and a Roundup-tolerant genetically modified maize” by a group of research scientists of the French Committee for Research and Independent Information on Genetic Engineering (CRIIGEN), an institutional member of ENSSER. (Seralini Gilles-Eric et al., 20120918) report on a two-year, full life-cycle study with rats testing Monsanto’s NK603 glyphosate-tolerant GM maize (single trait glyphosate tolerance) and its Roundup co-technology in the journal Food and Chemical Toxicology. The group of researchers led by Prof. Séralini has published previous toxicology studies on Roundup and its active ingredient glyphosate ((Gasnier, C. et al., 2009); (Benachour & Seralini, 2009) The scientists also evaluated historical industry data. These data from feeding trials with rats were submitted in support of the company’s dossers seeking approval for food/feed import. When reanalysing the developer’s raw data, they found signs for toxicological effects on rat liver and kidneys after 90 days of feeding with GM maize, including the GM maize NK603 tested in this new study ((de Vendomois et al., 2009; Seralini et al., 2011; Seralini, G.E. et al., 2007; Seralini, G. E. et al., 2007).

Repeated calls that regulators should require more rigorous, long-term follow-up studies by the developers have consistently been ignored or rejected on dubious grounds. In 2011, the European Food Safety Authority (EFSA) still rejected mandatory 90-day feeding studies as a requirement for applications for GM food and feed (EFSA Opinion, 20110524). The few studies that were carried out by developers are voluntary and apply protocols of their choosing.

Main issues according to ENSSER

1. CRIIGEN’s research1 was crucial in finally eliciting a policy response by the European competent authority, the European Commission’s Directorate-General SANCO (Health and Consumer Safety) in 2012. In its draft Implementing Regulation on applications for authorisation of genetically modified food and feed (European Commission 2012), the European Commission stated: “toxicological studies with the whole genetically modified food and feed shall be carried out.” If adopted, applicants “shall include a 90-day feeding study with whole food and feed in rodents for the assessment of food and feed containing, consisting of or produced from genetically modified plants [...].”

2. After a careful comparative analysis of both industry published data and that of CRIIGEN, ENSSER concludes that most arguments which attempt to invalidate the Séralini et al. study cannot hold up to closer scrutiny. Raised criticisms are to a large extent either wrong or apply double standards. The weak point of the pilot study by Séralini et al. is the number of animals used, which does not allow a statistical analysis of the raw data regarding one parameter measured out of over 30 - mortality. This has been acknowledged by the authors and needs to be considered/remediated in follow-up studies.

3. The controversy and vitriolic attacks evoked by the study reveal one underlying aspect: The lack of appropriate and agreed methodologies for long-term studies to scientifically assess effects of the life-time consumption of GM foods.

4. The development of such methodology and tests, which have been demanded by concerned scientists ever since GM food was announced to be introduced into the international markets, has systematically been blocked by lobby work of industry and associated scientists. International bodies such as the Codex Alimentarius and national governments - including most EU governments and their authorities - accepted instead the concept of substantial equivalence and the concept of familiarity to evade any scientific mandatory testing of GM food for human health safety and to simply declare significant differences found between GMOs and their unmodified parents as ‘biologically irrelevant’ in an assumption- based approach.

5. The acceptance of these industry-led constructed models providing the conceptual justification for evading testing of food-related risks associated with the introduction of this new technology and neglecting the clearly formulated demands of European citizens led to the crisis of trust in science & regulations that now come to light with full force.

6. Due to the proven close links between industry and EU risk assessors and the documented disproportional influence on regulations by developers and owners of the technology, it is predictable and expected that these authorities, including EFSA, will not be able to substantially revise their original assessment of GM maize NK603 (and any other application) as their credibility is at stake. This is highlighted for example by the European Parliament (2012) refusing the discharge of the EFSA 2010 budget as long as there is no fundamental change in policy, leadership and guidance.” (ENSSER Statement Seralini, 20121005)

Just four points of the ENSSER defense commented here:

Point 1: (Point 3 in ENSSER): ENSSER and Séralini admit the weak point of the study: The low number of tested animals: “The weak point of the pilot study by Séralini et al. is the number of animals used, which does not allow a statistical analysis of the raw data regarding one parameter measured out of over 30 - mortality. This has been acknowledged by the authors and needs to be considered/remediated in follow-up studies.”

Strangely enough, Séralini himself defends at roughly the same time the low number of animals in this answer to critics (Seralini, Mesnage, et al., 2012) 14

“We thus biochemically measured 10 rats per sex per group as performed by Monsanto. Even for a study of up to two years, we had no reason to monitor biochemical effects on more than 10 animals per sex per group as this is the number recommended in OECD guideline 452 for chronic toxicity testing (OECD 1981 was in application when the study started in 2008), even if 20 animals per group or more are possible. The purpose of the addition of R treated groups was not to assess R long term carcinogenesis, which needs to follow OECD 453 guideline with at least 50 rats per sex per group (even if 10 rats are then still measured at a biochemical level). The aim of our study was to test R under similar conditions to the GM maize in order to try and understand if residues of R in the feed could explain the possible pathologies that may arise. There were two main potential sources of harm tested in our study: (i) effects from the GM maize itself, treated or not with R, and (ii) herbicide residues alone in drinking water, using 3 doses for each treatment. We recall that the initial investigation published by Hammond and colleagues (Hammond et al., 2004) used 2 doses for each treatment group despite that fact that 3 doses are recommended by OECD guideline 408, which they reported to have followed.” (Seralini, Mesnage, et al., 2012)

Point 2: Lack of quantitative feed details given in his long term study, cheap and false excuse by claiming that Hammond in his papers also lacks precise quantitative data on rat feeding:

ENSSER thus proves that they did not exactly read the two Hammond publications of the experiments: (Hammond, B. et al., 2004; Hammond, B. G. et al., 2006): ENSSER claims that Hammond did not give details about the amount of feed, relying on the second publication from 2006, but actually Hammond refered in the 2006 publication explicitly to the 2004 publication, where all details, including a graph, are given.

Point 3 (Point 6 in ENSSER) Independency of EFSA: EFSA is an European Agency with probably the strictest hiring rules existing, conclusion: it’s as independent as as it can possibly be: (EFSA, 20121011; EFSA Independence, 20121028b)

EFSA Independence (20121028), On the Indepencence of EFSA, Collection of 32 website links on the Independence of EFSA, publ: EFSA, Parma, http://www.youtube.com/watch?v=N0FEVglpQcw&feature=player_embedded AND http://www.efsa.europa.eu/en/topics/topic/independence.htm AND http://www.ask-force.org/web/Seralini/EFSA-Independence-Statements- 20121028.pdf AND http://www.ask-force.org/web/EFSA/EFSA-response-NGOs-EN-2012.pdf AND http://www.ask-force.org/web/EFSA/ILSI-EFSA- response-NGOs-EN-2012.pdf AND http://www.ask-force.org/web/EFSA/Scott-Thomas-EFSA-denies-conflict-interest-allegations-20131025.pdf

And

EFSA (20121011), European Court of Auditors recognises EFSA’s advanced independence policy, makes recommendations, publ: http://www.efsa.europa.eu/en/press/news/121011.htm AND http://www.ask-force.org/web/Seralini/EFSA-Cour-Autitors-Recommendations-20121011.pdf

Additional remarks: It is a strange way of looking at independency when you know that oppositional groups are often financed by organic industry units, see (Olenyi Sebastian, 20120228)

Olenyi Sebastian (20120228), GM crops: Forget the money, follow the science publ: The BTS blog of the Biotechnology and Society group of the TU Delft and Euractiv, http://www.euractiv.com/cap/gm-crops-follow-money-analysis-510724#comment-2945 AND http://www.ask- force.org/web/Fundamentalists/Olenyi-GM-Crops-Forget-Money-Follow-Science-20120228.pdf and (Seppi Wackes et al., 2011)

Seppi Wackes, Imposteurs and C. (various) (2011), Seralini c. Fellous : la science politisee, la justice instrumentalisee, les masques tombent ! (1,2,3.,4,5), par Wackes Seppi in: Imposteur, Wackes Seppi, http://imposteurs.over-blog.com/pages/Tout_ou_presque_sur_le_CRIIGEN- 4536267.html AND http://imposteurs.over-blog.com/article-seralini-c-fellous-la-science-politisee-la-justice-instrumentalisee-les-masques-tombent- 2-par-wackes-seppi-65324096.html AND http://imposteurs.over-blog.com/article-seralini-c-fellous-la-science-politisee-la-justice-instrumentalisee- les-masques-tombent-5-par-wackes-76194721.html http://www.ask-force.org/web/Seralini/Testar-Temoignage-Seralini-20101124.pdf AND http://www.ask-force.org/web/Seralini/Joudrier-INRA- Seralini-Rebuttal-french-230501.pdf AND http://www.ask-force.org/web/Seralini/Fellous-Seralini-Jugement-20110118.pdf AND http://www.ask- force.org/web/Seralini/Haro-von-Mogel-blog-Seralini-2011.pdf AND http://www.ask-force.org/web/Seralini/Doull-Seralini-Report-2007.pdf AND http://www.ask-force.org/web/Seralini/monsanto-backgrounder-richar-response-2005.pdf AND http://www.ask-force.org/web/Seralini/Fellous- Seralini-Jugement-20110118.pdf

Point 4: The Séralini defense of ENSSER makes a false claim by stating that the Sprague-Dewey rats were used for similar experiments successfully and without complaints, and in order to underpin this statement, ENSSER cites several papers underpinning this opinion:

ENSSER cites 12 long-term experiments with Sprague-Dewey rats showing no problems: This is a false claim, since not a single of those 12 ENSSER citations support their hypothesis, all authors without exception point to the difficulties of a clear distinction between spontaneous or experimentally induced carcinogenicity of body organs. There are only two possibilities: Either the authors of the ENSSER rebuttal did not really download the full texts of their citations and in some cases did even not read the summaries, or they counted on readers who do not invest the time to read the citations properly.

 (Voss et al., 2005) have used 600 rats resulting into an incontestable significant significant dose-related trend, testing phthalates, and concluding explicitly that Sprague-Dewey rats are not suitable for those long term experiments. “In addition, the carcinogenicity should be evaluated in rat strains with low spontaneous tumor incidence in the organs known as target of DEHP toxicity”

15

 (Hack et al., 1995) calculate proper statistics from groups of 50 rats, testing insecticides (Endosulfans), detrimental toxicological effects proven, but no carcinogenicity (no surprise). “The type and frequency of tumours were similar in dosed and control rats and gave no indication of a treatment-related effect.”

 (Klimisch, Deckardt, Gembardt, Hildebrand, Kœttler, et al., 1997), the same story with a toxicity study on N-vinylpyrrolidone-2 vapours (NVP): Sprague rats seem to be unsuitable for long time carcinogenic studies: “As NVP gave negative results in a battery of in vitro and in vivo genotoxicity tests, it appears that the tumours that arose were manifestations of a non-genotoxic mechanism.”

 (Minardi et al., 2002): A special case of Vinyl acetate monomer (VAM) causing superficial cancer symptoms (squamous cell carcinomas), there where VAM came into direct contact with surfaces in the oral tract, but again the use of Sprague rats proved to be inadequate for general toxicological long term studies. “The experimental studies on rodents conducted until 1997 proved in one way or another inadequate to evaluate the carcinogenic potential of VAM. To date, seven carcinogenicity studies have been published in the scientific literature. In the first study, 96 male and female Sprague-Dawley rats were exposed to 2500 ppm VAM by inhalation for 52 weeks. Early mortality was high: only 49 animals survived for 26 or more weeks. No tumours related to VAM were reported during 135 weeks.15–17 Because of the poor survival rate, this study was inadequate for detecting the carcinogenic potential of the monomer

 (Soffritti et al., 2006) Again the same story: the induced tumours not statistically confirmed: “In our experimental conditions, it has been shown that sodium arsenite administered for 104weeks in drinking-water to male and female Sprague-Dawley rats, 8 weeks old at the start of the experiment and kept under observation until spontaneous death, induces an increased incidence (albeit not statistically significant) of benign and malignant tumours of the lung, kidney, and bladder.”

 (Lee et al., 2010) demonstrates the existence of a rare tumor incidence at early age of the Sprague rats, certainly not a good prospect for tumor studies with this rat strain. Spontaneous basal cell carcinoma (BCC) is very rare in rats, with an incidence rate of only 0.14% reported in aged animals. “A spontaneous BCC occurred in a 7-week-old Sprague-Dawley rat housed in a specific-pathogen-free animal facility. The tumor was a single, well-delineated reddish-brown subcutaneous mass measuring 2 _ 2 cm and located in the left inguinal region. Microscopically, the tumor consisted of basaloid cells in lobular and cribriform growth patterns and with a high mitotic rate. Immunohistochemically, cytokeratin 14 (an indicator for basal keratinocytes of the epidermis) showed strong reactions throughout the whole tumor, and cytokeratin 18 showed weak but positive reaction in the majority of nested tumor cells. To the authors’ knowledge, this is the first report of spontaneous BCC occurrence in young Sprague-Dawley rats.”

 (Perri et al., 1981) reports about a 30 months experiment (not “Perry et al. 1981 as a 12-months study” – not existing…), showing only at an unreal high dose of the yeast added to the feed some tumours symptoms: “Animals dead spontaneously or killed at the end of the trial were autopsied and the main organs fixed for histological examination. Lesions and tumours were classified. Biochemical, haematological and autopsy variations at the times of sacrifice were observed in the long term study. The experiment showed no pathological differences between controls and animals treated with 20 and 50% proteins from yeast. The group fed with 80% single cell protein showed a significant increase of malignant lymphomas incidence. ”And: “it is important that the experiments of this type should be evaluated with caution since it is difficult to separate the effect of possible nutritional imbalance from the effect of spontaneous ageing and the actual effects of some component of the diet. As a conclusion the results in our experiments show that the diets in which 18.4 and 7.2% are supplied by SCP [= Single Cell Protein] do not modify the distribution of tumours as compared to the control group.”

 (Conti et al., 1988, online 2006) A 12-months study showing slightly better results for testing Syrene oxide, because the Sprague rats life was terminated when the survival rate reached 50%, except for testing. Again the main symptoms were superficial cancer (squamous cell carcinomas). “The study was terminated when the survival rate reached 50% in at least one experimental group. Styrene, when given by inhalation, was found to cause an increase in total (benign and malignant) and malignant mammary tumours. Styrene oxide produced a high incidence of tumours in the forestomach (papillomas, acanthomas, and in situ and invasive squamous cell carcinomas). Para- methylstyrene was not shown to be carcinogenic.”

 (Maltoni et al., 1988) A 12-months study, from the same research group of B. Conti demonstrate the same symptoms of the inadequacy of Sprague rats for Long-term experiment: Unexplained differences occur between inhalation and ingestion: “Under the tested experimental conditions, acrylonitrile was shown to be carcinogenic in Sprague-Dawley rats when given by inhalation and did not produce any carcinogenic effect when given by ingestion. In the inhalation experiment, acrylonitrile caused an increase in different types of tumours and the most noticeable acrylonitrile-related tumor was encephalic glioma.”

 (Morcos & Camilo, 2001), another 12-months study with no results on cancerogenicity of the texts substances, not surprising: “Acutely and chronically, there were no differences in external appearance, level of activity, daily food consumption, blood cell count, kidney function, thyroid function, prothrombin time (PT), and activated partial prothrombin time. (PTT), which remained within normal ranges in the supplement group. Organ histology remained unchanged. Although during the chronic toxicity period the triglyceride and LDL suppression persisted,”

16

 (Flamm et al., 2003) in this 24-months study is not relevant for the cancerogenicity studies under debate here, there is no trace of tumor findings, and the conclusions fit well into the hypothesis of the experiment: “Lower B Wand BWG were the only consistent findings in neotame safety studies. These changes are consistent in magnitude with small reductions in FC during long-term neotame safety studies according to the allometric relationship between changes in BWG and changes in FC previously reported for other high-intensity sweeteners and dietary restriction studies in the same sex and strain of rats.

 (Guiterrez et al., 2011): The same results again: Sprague-Dewey rats are not suitable for tumor testing: “Four of 160 rats died before study completion. No clinical signs of toxicity were observed throughout the study. Food and water consumption, bodyweight, blood biochemical and haematological parameters, organ weight ratios and histopathological findings were similar in control and treated groups. The histological lesions found in treated animals are commonly present in this specie and strain according to literature and our historical data. In conclusion, long-term (12 months) oral treatment of rats with D-004 (800–2000 mg/kg/day) did not show evidences of D-004-related toxicity under our conditions. The highest dose tested (2000 mg/kg) was a no-observed adverse effect level in this study.”

KEENAN ADVERSE EFFECTS IN 1999

Apart from those above polemics, ENSSER provides some arguments in a scientific style which have to be commented here:

2.3.3. ENSSER AND SÉRALINI ADMIT MAJOR MISTAKE: INSUFFICIENT SAMPLE SIZE

The authors of the Seralini study acknowledge the inconclusive low number (10) of rat groups compared, which renders the whole study anyway inconclusive. Actually, the debate could be closed here for good, but Séralini et al. are not stopping here with their polemics.

The original wording of the former CRIIGEN president Corinne LePage in a dispute sounds a bit different: (LePage Corinne, 20120925), published a few days earlier, still defending stoutly the low number of test groups:

“The number of animals tested per group were insufficient” ... Except that no study has ever been conducted on as many animals (200) and the number of rats per group (10 [in Séralini’s case, it was 10 of each sex]) is the same as the number used in all studies submitted to public bodies and validated by them [GMW comment: Monsanto used 20 rats per group in its 90-day study on NK603 but reportedly only analysed 10!”

2.3.4. FEED UPTAKE NOT QUANTIFIED

By using flawed comparisons to Monsanto papers they intend to excuse their own omission of not having quantified the administered feed.

Again the same syndrome: ENSSER does not study the referenced literature of B. Hammond et al. from Monsanto thoroughly enough: Contrary to ENSSER, Hammond et al. give precise amounts of ingested feed in their publications, but for this you again have to read the paper thoroughly, not only the remark “at libitum feeding”, you need to include the references given by Hammond: It is correct that (Hammond, B. G. et al., 2006) write explicitly that the food has been administered ad libitum, but in the same paper they refer to an earlier publication that all feeding and methodological details are given there (Hammond, B. et al., 2004): The consumption there is precisely quantified, given in the graphs of fig. 2 in g/day over the study weeks:

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2.3.5. AD LIBITUM FEEDING: WELL DOCUMENTED NEGATIVE EFFECTS

Somehow unexpected for the author is the complete ignorance related to the Séralini comments and his own writings – even in the experimental concepts of the politically motivated repeat experiment - that ad libitum feeding has a long negative history, there are numerous experimental papers which give ultimate prove that ad libitum feeding results in overfeeding and higher negative effects.

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Fig. 1 The cumulative percentage of male SD rats with lethal pituitary tumours ●.-The cumulative percentage of male SD rats with lethal pituitary tumors.

2.3.6. NO WRITTEN STUDY PLAN APPROVED BY DATED SIGNATURE OF THE STUDY DIRECTOR EXISTING The authors in ENSSER admit freely, that they did not follow a basic rule in toxicology experimentation: To set up the methodology according to a clearcut hypothesis before you start your work, another fundamental mistake: “The authors did not apply the OECD Guidelines 451 carcinogenicity Studies) or 453 (Combined Chronic Toxicity\Carcinogenicity Studies) - as demanded by commentators - because the authors did not intend to conduct carcinogenicity studies in the first place. It was their intent to apply the Guideline 408 methodology in an extended time span.”

The principles of good laboratory practice adopted formally by the OECD in 1997 (OECD Principles, 1998) (not cited by ENSSER) demands a proper study plan:

“For each study, a written plan should exist prior to the initiation of the study. The studyplan should be approved by dated signature of the Study Director and verified for GLP compliance by Quality Assurance personnel as specified in Section 2.2.1.b., above. The study plan should also be approved by the test facility management and the sponsor, if required by national regulation or legislation in the country where the study is being performed.”

2.3.7. FINAL SUMMARY, A COMPARISON OF OMISSIONS AND FACTUAL STATEMENTS OF SÉRALINI ET AL. 2012

From a German lab toxicologist I received a table which I translated, the colors applied to the items, comparing the 90-day experiments with the 2-years trials with laboratory animals, using the two OECD guideline publications 19

If you follow the red and yellow remarks, you will see that the paper of Séralini critizised here is not following the basic principles of long term feeding trials.

A similar picture is produced by the following Table, published by (EuroBioNews, 20121012). 20

The commentary of EuroBioNews is very critical about Séralinis data: The data do not show a proper dose- dependent effect.(Ng Ashley, 20120925)

The original data of Séralini’s publication from September condensed into a graph by Prof. Dr. Klaus Jany, December 2012

Following these first quick rebuttals the publications of the same author gave more precises graphs and comments: (Jany Klaus, 2013, 20120921; Jany Klaus & Widhalm Kurt, 201305, 20130409), a summary:

A peer reviewed rebuttal of Séralini: (Jany Klaus, 2013) 21

Jany Klaus (2013), Critical remarks on the long-term feeding study by Séralini et al. (2012). Does the study provide proof of health threats posed by genetically modified foods?, EFFL - European Food and Feed Law Review, 3, pp. 176-187, http://www.ask-force.org/web/Seralini/Jany-Critical-Remarks- Seralini-Study-EFFL-201303.pdf

“In the opinion of CRIIEN and Prof. Seralini the results of a long term feeding study with rats and genetically modified maize and the herbicide Roundup formulations are so serious that they have chosen several PR-events to present this new publication. In press conferences Prof. Seralini explained his feeding study demonstrate the scientific proof that rats develop earlier immense large tumors and died earlier due to the consumption genetically modified maize and/or roundup. Therefore he and CRIIGEN were convinced of the necessity to inform politicians and the public about the risks and danger of genetically modified foods and the use of the herbicide Roundup in agriculture. Furthermore he stated his study shows clearly that the safety assessment of transgenic plants and foods derived from them is performed in a wrong way or is insufficient. He and his supporters claimed that the findings are so alarming that a ban on transgenic plants is justified until valid procedures and criteria for a risk assessment are developed. In addition they discredited regulatory / safety assessment authorities, especially EFSA. Strange but possibly by chance in the same time period Seralini presented his new book “Tous cobayes” (We all are guinea pigs) in which he describes the risks of gene engineering, the use of plant protection agents and genetically modified foods. Furthermore the food supermarket company Carrefour, a financial sponsor to CRIIGEN and Seralini announced the introduction of a pallet products which are free from components derived from of genetically modified organisms (GMO-free products). These foods should protect consumers for genetically engineered ingredients and should enable a choice of freedom. Probably if all these public events were not by chance, one can assume that the findings of the just published feeding study were used for a populistic propaganda story against gene engineering in agriculture and food production. The long term feeding study of Seralini et al. found great interest in media as well as to NGOs and science. Only driven by the reported results, critics and opponents of GMOs saw here the longed scientific evidence of the risks for human health when consuming foods derived from GMO. These products and especially transgenic plants are more than harmful. They also promote cancer diseases and led to an earlier death. Therefore such products must be forbidden. On the other site, most of the scientists were shocked that such a manuscript could be accepted to publication in a peer-reviewed journal like “Food and Chemical Toxicology”. In the paper the questionable study design and so many methodical insufficiencies are apparent and nearly no measured data are presented. Especially no data are given verifying the statements about increased cancer rates and earlier death. The confidence in the reviewer system and/or in the scientific expertise of the selected peer reviewers was undermined. More than 37 governmental food authorities and academic societies have evaluated of the few available data and the whole publication. After the assessment they concluded nearly with the same opinion: The performed study does not correspond to generally accepted scientific criteria and the paper and the results are not suitable for a (new) risk reassessment of maize NK603 and glyphosate/roundup.” (Jany Klaus, 2013)

Life time / mortality

“The authors describe quite succinctly that approx.50% of the male and 70% of the female animals die earlier before reaching the normal expected lifetime (2 years) compared to the control group. They interpret this observation as a clear proof of the toxicity of the genetically modified maize and Roundup and glyphosate, respectively. However, such a simple unreflected consideration is not valid. It includes neither possible dose-response relationships nor differences between the drinking water test group and the maize NK603 feeding groups. Furthermore, the reference points in the control group have also to be regarded in respect to mortality and life time of the females. Seralini et al. observed only two deaths in this control group. This number is very low or in expressed in another way the survival time of the whole female test group is with 80% exceptionally high. The historical values for this rat strain are in the range of about 40-50%. Probable the observed high survival rate is purely by chance or arises from the too low number of animals used in the control group. Mortality and survival times of the animals in the different test groups should be analyzed separately. Derived from Sealini’s data the survival times of the males and females are here presented a little bit clearer in Fig. I A and I B).” (Jany Klaus, 2013).

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Fig. 2 Fig. IA: Survived animals in the drinking water test group from (Jany Klaus, 2013) See the 2250 mg/L male line , obviously, the high glyphosate concentration has some cancer protective characteristic…

“A negative influence of glyphosate is hardly to detect over the male test group. Mortality and life times are not significantly different from the control group at concentrations of 50ng/L and 400 mg/L. In contrast, the highest concentration (2.25 g/L) has positive effects. Fewer animals die and survival times are expanded. A prolongation time of almost 100 days can be observed (Fig IA). Obviously, high amounts of glyphosate also seem to counteract carcinogenesis. If one accepts this observation as a fact, then one must assume that glyphosate (Roundup) has protective health effects for male rats. However this finding would be in contrast to other studies of Seralini. Apparently females in all three test groups die earlier than in the control group. However, no dose-effect relationships are detectable at the different glyphosate concentrations. The observed lower survival times are probably due to the surprisingly high life times compared to the control group. Interestingly the survival times of the treated groups correspond essentially to the historical data for “untreated” rats. Obviously the survival times of this rat strain is not negatively correlated with the intake of glyphosate (Roundup). The small differences between the 3 test groups are not significant. (Jany Klaus, 2013)

Fig. 3 Fig. IB: Survived animals in the NK603 feeding test groups from (Jany Klaus, 2013)

Fig. 4 Fig. II Numbers of tumors found in the maize feeding test groups from (Jany Klaus, 2013)

Comment to Fig. I and II: 23

“Raw data can be taken from Fig. 2 of the Seralini study about the numbers of tumors of non-regressive palpable tumors and their temporal appearance. The most frequently observed anatomical pathologies are represented in Table 3. In Fig 2 it is shown that in both control groups no tumors were observed during the first 420 days, while already 10 - 30 % of treated female animals have developed tumors. On Fig. IB: Survived animals in the NK603 feeding test groups the other hand, tumors were detected not before 520 days in all male test groups with Roundup in the drinking water. Therefore it is not very informative to discus the occurrence and numbers of tumors in general over all the different test groups, but again it is conspicuous that there are no dose effect relationships observable. The lower the percentage of maize NK 603 or Roundup the earlier tumors were developed, whereas the highest concentrations apparently prevented an early development of tumors. As only three doses are tested an inverse dose-effect-relationship is not proofed. However, the data have to be analyzed separately and sex- specifically. As already observed in the drinking water test groups females seem to be more sensitive to genetically modified maize (maize NK603 with and without Roundup) due to the expanded life time females However, this increased sensitivity is not specific for genetically modified products (foods) or roundup formulations, but it is also observed when testing conventional, single substances. Within the individual test groups the overall numbers of tumors do not vary insignificantly compared to the control groups (Fig. II). In the male control group 13 tumors were observed, where as in the test group fed with maize NK603 (11, 22, 33%) 15, 13, 17 tumors were detected. In the test group fed with maize NK603 treated with Roundup less tumors (9, 6, and 10) were found than in the control group. The number of large tumors varies between 1 and 3, as shown also as a photo. Not surprisingly in females more tumors occurred. In the control group 19 tumors were detected. In the test group fed with maize NK603 26, 19, 25 tumors were reported. The number of large heavy tumors varied between 6 (control) and 10 (33% maize NK603). These slight differences can not regarded as significant for the occurrence of tumors over all test groups. Furthermore, the earlier appearance of the tumors can not proven by the Fischer-Kaplan statistic. The published data do not support a dietetic influence on the number of tumors or the earlier appearance of tumors.” (Jany Klaus, 2013)

Equally important is the rebuttal of Séralini from Spain: (Arjó et al., 2013):

Arjó, G., M. Portero, C. Piñol, J. Viñas, X. Matias-Guiu, T. Capell, A. Bartholomaeus, W. Parrott and P. Christou (2013), Plurality of opinion, scientific discourse and pseudoscience: an in depth analysis of the Séralini et al. study claiming that Roundup™ Ready corn or the herbicide Roundup™ cause cancer in rats, Transgenic Research, pp. 1-13, http://dx.doi.org/10.1007/s11248-013-9692-9 AND http://www.ask-force.org/web/Seralini/Arjo-Plurality-Opinion- Scientific-Discourse-Seralini-2013.pdf AND http://www.ask-force.org/web/Facultyof1000/Arjo-Plurality-Opinion-Seralini-F1000-Ammann-20130311.pdf

“A recent paper published in the journal Food and Chemical Toxicology presents the results of a long-term toxicity study related to a widely-used commercial herbicide (RoundupTM) and a Rounduptolerant genetically modified variety of maize, concluding that both the herbicide and the maize varieties are toxic. Here we discuss the many errors and inaccuracies in the published article resulting in highly misleading conclusions, whose publication in the scientific literature and in the wider media has caused damage to the credibility of science and researchers in the field. We and many others have criticized the study, and in particular the manner in which the experiments were planned, implemented, analyzed, interpreted and communicated. The study appeared to sweep aside all known benchmarks of scientific good practice and, more importantly, to ignore the minimal standards of scientific and ethical conduct in particular concerning the humane treatment of experimental animals.” (Arjó et al., 2013)

The recommendations:

“We do not advocate retraction of scientific papers simply in response to public pressure. However, the publication of the Se´ralini article was a clear and egregious breach of the standards of scientific publishing and a grave insult to the integrity of thousands of dedicated scientists around the world. We therefore call upon the editor of Food and Chemical Toxicology to issue formal retraction of the Se´ralini article, at the very least in order to comply with COPE guidelines. We note that simply publishing letters to the editor pointing out the deficiencies with the paper does not relieve the editor, the journal or its publisher from the need to abide by COPE guidelines. The publication of the Séralini article undermines the value of peer review, encouraging the plurality of opinion and democracy in science and promoting their influence on scientific policies. Therefore, we also recommend that all journal editors abide by COPE guidelines in order to protect the credibility of the peer review process and protect the integrity of the scientific process in the public view. Although COPE guidelines should be sufficient for proper editorial decisions, we also call upon all editors to recognize that their decision to publish certain papers can have serious repercussions, including the introduction of unnecessary new regulations, the escalation of expenditure in the search to ensure compliance, the unfair suppression of promising technologies and unnecessary alarmism that affects the most vulnerable members of our society.” (Arjó et al., 2013)

3. SECOND SERALINI STUDY IN PLOS ONE, STATUS 20150729

Here you can compare the pre-release leaked through his buddies (same scandalous behavior as in the press conference 2012, with the exception that this time he released all original data). But being able to compare both versions, the final release by PLOS one with some changes, which are considerable, but strangely enough incomplete, it again smells like sloppy peer revive for many reasons.

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We all are a bit disappointed since we thought PLOS one is retracting the paper before the official publication, this would have been a world premiere for Séralini…

Comparison document: (Windows 2013, go to REVIEW and fourth left to COMPARE and choose your preferred constellation under Markup: http://www.ask-force.org/web/Seralini/Mesnage-Seralini-COMBINED-final-preprint-1.docx

The first (illegal) release to his friends – in these leaky times easy to get – those who were quick enough got it directly on the link in the press conference June 17 (Details under remarks in the first press release). http://www.ask-force.org/web/Seralini/Mesnage-Defarge-et-al-PlosOne-20150617.pdf

The final (official) release from PLOS one from July 2: http://www.ask-force.org/web/Seralini/Mesnage-Defarge-et-al-PlosOne-final-20150702-1.pdf

The press release from June 17, 2015 http://www.ask-force.org/web/Seralini/Mesnage-Defarge-et-al-Test-Rat-Diets-PlosOne-Press-Release- 20150617.pdf A juicy detail: in the June 17 press release the link “Published in PLOS ONE: http://dx.plos.org/10.1371/journal.pone.0128429 (17 June 2015)” did not function ca. from June 18 on, now it works again, leading to the document published today July 2. The reason: unexpected instruction from PLOS one that original version is withheld and needs correction.

Another press release, basically with the same false pre-release proclamations. http://www.ask-force.org/web/Seralini/Mesnage-Defarge-et-al-Test-Rat-Diets-PlosOne-Press-Release- UCBN-20150617.pdf

The details under the COMPARISON document above: PLOS one demanded to cancel some sentences before release: The major cancelled text in the summary:

The high background rate of pathologies in laboratory rodents could be due to dietary contaminants. This invalidates the use of external controls (historical data) in regulatory tests consisting of comparisons of toxicological effects of control rats from other experiments, because those rats are fed with different mixtures of pollutants. This also questions the use of 50 rats per group in carcinogenicity studies to increase the statistical power lost due to the elevated pathology background.

In the Introduction cancelled: To estimate the hazards due to chronic exposure to these contaminants in diets we calculated the chronic non-cancer hazard indexes (HI) (to take into account general toxicological

Strangely enough, in the conclusions at the end of the paper nearly the same statements which they had to cancel in the summary are again repeated.

As far as I can understand as a non-Toxicologist, there is not the slightest hint that the measured pesticide components really can harm the rats or rat-experiment in ANY WAY. It’s just an unproven song through the whole document, and for sure it will create fear again with non-specialists. My French friends created Séralinade 1 and 2

You can only guess what this means: most probably the lazy reviewers did not arrive to the end of the text, or relied on the authors to wipe out similar statements, WHO KNOWS?

Then I tested citation No. 69 as an example with a result typical for Mesnage-Séralini: Kuroiwa, Y., Ando, R., Kasahara, K., Nagatani, M., Yamakawa, S., & Okazaki, S. (2013). Transition of Historical Control Data for High Incidence Tumors in F344 Rats. Journal of Toxicologic Pathology, 25

26(2), pp. 227-230. http://www.ask-force.org/web/Seralini/Kuroiva-Transition-Historical-Control- Data-High-Incidence-Tumors-F344-rats-2013.pdf

The comment of Mesnage et al on this reference 69 is: “Kuroiwa et al. [69] concluded that the diverse and fluctuant incidences of pathologies in historical data may be caused by environmental factors, rather than “spontaneous”

But when you really read Kuroiwa, you see not the same conclusions, re environment it is just the CONTRARY. “Our series of surveys demonstrated that the historical control data for spontaneous tumors fluctuated over the years. In our survey, all the rats came from the same breeder, and were housed under the same laboratory conditions. The fluctuation of tumor incidences might be caused by some sort of factor from animals other than environmental factors, although genetic drift is rarer in the inbred F344 strain than in the outbred Wistar and SD strains see

Tennekes, H., Kaufmann, W., Dammann, M., & Ravenzwaay, B. v. (2004). The stability of historical control data for common neoplasms in laboratory rats and the implications for carcinogenic risk assessment. Regulatory Toxicology and Pharmacology, 40(3), pp. 293-304. http://www.sciencedirect.com/science/article/pii/S0273230004001060 AND http://www.ask-force.org/web/Seralini/Tennekes-Stability-historical-control- data-neoplasms-2004.pdf )

It is a real crux when you start reading papers thoroughly… Also Koroiwa cites a paper stating that the SD rats have much lower tumor incidences, and when you go to the paper, it does not at all deal with SD rats. (Besides this, the statement is dead wrong as has been shown by multiple publications, SD rats are simply not compatible with long term experiments due to early appearance of spontaneous tumors.

Some final remarks: Séralini intends to construct two excuses for mistakes in his previous papers: 1. The number of samples too low (which he actually admitted), but now comes back with a really cheap excuse: fluctuation due to different pesticides in the feed a higher number of samples is useless…. 2. SD rats justified, despite a many times proven high rate of spontaneous tumors of various kinds.

Presently, I am analyzing nearly ca. 190 full-text references on experimental use of Sprague – Dawley (SD), the case is clear, in the great majority of the cases the SD rats are highly prone to spontaneous tumors in early age and should not be used for long term experiments.

Did you know that Séralini himself admitted that having reached mid-term on the experiment in 2011/12, he decided to change from short term to long term concept?? This guy is chaotic and highly selective in experimentation and reading papers.

One excellent reaction is already published, since then we have even more ammunition

Bernstein Alison. (20150618). Scientist deconstructs Séralini’s PLOS GMO study: ‘Failed attempt at redemption’ with 54 comments. Genetic Literacy Project, 20. http://www.geneticliteracyproject.org/2015/06/18/scientistdeconstructsseralinisplosgmostudyfailedattemptatredemption/ AND http://www.ask-force.org/web/Seralini/Bernstein-Scientist-deconstructs-Seralini-PLOS-study-20150618.pdf

For some time, we considered to write a letter to PLOS ONE, but decided that the paper did not stir up enough dust and it might be better to avoid big noise.

Additionally some short comments on: IARC Study final, 29. 7. 2015, 92 pages http://www.ask-force.org/web/HerbizideTol/IARC-Glyphosate-Review-.July-2015.pdf 26

An excellent deconstruction of the Séralini PLOS ONE paper has been published by Alison Bernstein in GLP:

Bernstein Alison. (20150618). Scientist deconstructs Séralini’s PLOS GMO study: ‘Failed attempt at redemption’ with 54 comments. Genetic Literacy Project, 20, New York, GLP Genetic Literacy Project, http://www.geneticliteracyproject.org/2015/06/18/scientist- deconstructs-seralinis-plos-gmo-study-failed-attempt-at-redemption/ AND http://www.ask-force.org/web/Seralini/Bernstein- Scientist-deconstructs-Seralini-PLOS-study-20150618.pdf

The full text is given here as a follow-up text of the author’s rebuttal with some selected elements of critique, here a full rejection, including lots of general critical points about Séralini’s and his group’s work:

Gilles-Éric Séralini, a molecular biologist at the University of Caen in France, is hoping for redemption with a new paper about the effect of pesticides and genetically modified (GMO) feed on rats and mice. He hasn’t earned that redemption.

A few years ago, Séralini suffered the ultimate humiliation for a scientist. The JOURNAL FOOD AND CHEMICAL TOXICOLOGY retracted his high-profile study. The editors reviewed the raw data and found the results were “inconclusive” and did not back the conclusions that were loudly trumpeted in media headlines. The authors themselves eventually conceded that the study had serious flaws, noting in a press release that “the data are inconclusive, due to the rat strain and the number of animals used.”

Other long-term studies, which were publicly funded, had uncovered no healthissues with GMO corn or the herbicide glyphosate. The Japanese Department of Environmental Health and Toxicology released a 52- week feeding study of GM soybeans in 2007, finding “no apparent adverse effect in rats.” In 2012, a team of scientists at the University of Nottingham School of Biosciences released a review of 12 long-term studies (up to two years) and 12 multi-generational studies (up to 5 generations) of GM foods in the same journal that published the Séralini paper, concluding there is no evidence of health hazards.” Consequently, there was growing pressure on the journal to retract the original study since publication in 2012, along with other criticisms and an exchange of letters in the journal.

Now, Séralini has a new study, released July 2 after being delayed more than two weeks, in which the authors measured the levels of various pesticides, industrial chemicals and genetically engineered crops in 13 brands of laboratory rodent feed. (NOTE: An earlier version of the embargoed journal article had been distributed to journalists and numerous news outlets, and Séralini’s own vanity site has broken the embargo.) There seems to be nothing wrong with the data itself. The results themselves of are not surprising given the currently planted genetically engineered crops and current pesticides usage.

GLP has assessments from the latest Séralini study by scientists from around the world here.

GLP has a profile of Séralini and his research here.

The authors take these unsurprising results and call into question the validity of everyone else’s studies. The conclusion, as explained in a university press release, is unfounded: “It therefore appears that the cause of diseases and disorders found in laboratory rats has been too quickly attributed to the genetic characteristics of the species used.” In other words, the rats are dying from GMOs and pesticides like glyphosate in feeds.

As a neuroscientist who works regularly with lab animals, I find these claims baffling. 27

These results do not mean much, given the absence of any data suggesting a correlation between diet and phenotype, a trait, in laboratory rodents. They have presented zero evidence from their own work or published work that feed contamination is an issue for laboratory animal health. They present no data on animal health and no data about which feeds, fed to which strains produce which pathological phenotypes. They also completely ignore the fact that different strains of rodents have different phenotypes and rates of spontaneous pathology. The authors have made a huge logical leap in concluding that this data calls into question all historical data used as external controls.

Fortunately, the very historical data that the authors are attempting to discredit are unlikely to support their conclusion.

I would also argue that if such changes in the phenotype of well-characterized strains of lab rodents were occurring, the scientists who work with these animals would notice. When your control animals don’t behave as expected (in their behavioral response, pathology, life span, reproductive success, anything really), this is a big red flag. If there was an issue with feed causing significant pathology in laboratory rodents, this is something that would be seen in labs around the world.

If we wanted to mine the existing data to address some of these concerns, here are a few question we can ask to explore if these chemicals or genetically engineered crops in laboratory feed affect the phenotype of laboratory rodents.

When genetically engineered crops were introduced in the 1990s, did control mice start having different phenotypes? We can pull the rates of spontaneous pathological effects from papers published before and after the introduction of the relevant genetically engineered crops and compare them. If there is no difference in control animals before and after the introduction of genetically engineered crops, then the GMO composition of laboratory feed has had no effect on the health of lab animals.

A study published last year by University of California-Davis animal geneticist Alison Van Eenennaam did just that with livestock. Her team examined almost 30-years of livestock studies, more than 100 billion animals, comparing their health before and after GMO feed became the norm. She found no difference in the animals.

1) Some commonly used lab strains have been used for 70+ years. The pesticide residues found in laboratory feed reflect the pesticides in current use, just as pesticide residues on food reflect current pesticide use. So an obvious question is: have the phenotypes of these mice changed as our pesticide use has changed?

When organochlorines were banned in the 1970s and replaced by organophosphates, was there a concurrent change in the phenotype of lab animals? As use of organophosphates has declined more recently, have we seen a change in the animals? As glyphosate has replaced more toxic herbicides have we seen a change in lab animal health? Again, we can assess this by comparing control animals in different decades. If the phenotype of control animals has been consistent, these changes in pesticides residues found in laboratory feed have had no effect on their health outcomes.

2) In the paper, the two feeds used in Italy had the highest amount of contaminants, according to the authors. This leads to the question: do lab animals in Italian labs have a different set of phenotypes as a result of eating this specific brand of feed? A more general way to put that question is to ask if the same lab strains fed feed with different contaminant profiles have different phenotypes? 28

We can easily mine existing data to address these questions. Let’s look at Sprague-Dawley rats as an example to answer the first two questions. These rats were originally bred in 1925 so they have been used experimentally for enough time to answer questions 1 and 2 above. If we look at historical and current data on these animals, we can see if there have been any changes in their background phenotype. This has already been addressed in a rebuttal letter to a previous paper from Séralini’s group. The rate does not appear to have changed.

This specific breed of rats is well known to be prone to develop cancer with age and especially when there is no dietary restriction. For example, Prejean et al. (1973) noted a spontaneous tumour incidence of 45% in 360 Sprague–Dawley rats (179 males and 181 females) in an 18-month series of carcinogenesis experiments. The percentage of female rats with tumours was almost double that of males. Durbin et al. (1966) reported a mean incidence of 71%, the peak incidence in normally aging rats were age-related with abrupt increases in the rate of development of mammary tumour, one occurring at about the 500th and the other at about the 660th day of life, with the median age at 671 ± 41 days. Harlan, the company that marketed the animals, describes the high incidence of 76% of mammary gland tumours (predominantly fibroademonas) in females on Life-span and Spontaneous Disease of Sprague-Dawley. Keenan et al. (1995) describes spontaneous tumours in up to 87% of females and up to 71% of males fed ad lib. Dietary restriction significantly reduced the incidence of tumours.

3) To address the third question, we can compare data on control animals from studies done in different countries. Of course, the proper way to address these issues is to do a thorough meta-analysis of all control animals in studies, separated by strain. However, quick reviews of the literature for the incidence of spontaneous pathology doesn’t seem to justify such an effort. Basically, if changes in phenotype of commonly used strains of laboratory rodents were occurring, we would see it in the existing data. If adverse events were occurring in control animals, a properly conducted study would report these adverse effects and we would see these changes in behaviour of control animals in the literature. There are also ethical and legal obligations for reporting such adverse events to the veterinary staff and institutional ethics boards for animal research. Furthermore, the question of why the control mice are behaving differently than all other control mice would be very interesting research question that scientists would follow up on.

The authors of this study are searching for an answer to a non-existent problem. All Séralini had to do is a literature search to determine if this is a problem. They are trying to blame external factors (chemical contamination and GMOs) for a problem of genetics. We can, in fact, address many of the issues that authors say cannot be addressed by mining the very data that Seralini and colleagues want to throw away out of hand. But they didn’t address them and thus cannot make any conclusion except that genetically engineered crops and low levels of these chemicals exist in laboratory feed.

The authors also conclude that because of the high background pathology (reminder: not demonstrated here), the recommendation to study larger groups of animals is invalid. This is not how statistics works. If your variation is higher, you need a larger group to discern a pattern. If there is, in fact, high background variation, the only thing this underscores is the important of choosing an appropriate mouse strain for your study.

Overall, this paper is a thinly veiled attempt to address the consensus scientific criticism of Séralini’s previous work. This new paper doesn’t do anything to help his case that these criticisms were not valid. 29

Alison Bernstein is a scientist studying Parkinson’s disease living in Atlanta with her husband, 2 kids and 2 cats. Follow her on her Mommy PhD Facebook page and on Twitter @mommyphd2.

A rebuttal of the Seralini PLOS ONE study comes from EFSA (European Food Safety Authority) (EFSA, 20151006):

EFSA. (20151006). Review of results published by Mesnage et al. (2015) in PLoS ONE and the laboratory findings communicated by Dr Samsel(EFSA, 20151006) to Farm Wars. European Food Safety Authority. EJ EFSA Journal Scientific Report, pp. doi:10.2903/j.efsa.2015.4258 AND http://www.efsa.europa.eu/sites/default/files/scientific_output/files/main_documents/4258.pdf AND http://www.ask- force.org/web/Seralini/EFSA-Review-Mesnage-PLoS-ONE-and-Laboratory-findings-Samsel-Farm-Wars-20151002.pdf

Abstract In a paper published in PLoS One and entitled ‘Laboratory rodent diets contain toxic levels of environmental contaminants: Implications for regulatory tests’, Mesnage et al. (2015) analysed commercial laboratory rodent diets for environmental contaminants and genetically modified organisms (GMOs). In samples from 13 different commercial rodent diets obtained from five continents, the authors of the study report the presence of pesticides, heavy metals, polychlorinated dibenzo-p-dioxins and dibenzofurans, and GMOs. The paper by Mesnage et al. (2015) provides a useful addition to the already existing knowledge in the field. However, there are several limitations with the methodological approach used by the authors, including insufficient information about the test material and methodology used, incomplete reporting of the data, and inappropriate interpretation of legislation and results. The vast majority of pesticides were absent (below the limit of detection), and where detected, the levels of pesticides, heavy metals and dioxins were only just above the limit of detection in the feed samples but below regulatory levels for feed and foodstuffs. Only in a limited number of feed diets did the authors report levels of lead that exceeded the maximum levels specified by legislation for foodstuffs. The application of the ADI concept to claim the existence of a health risk in rodents or to demonstrate background levels of diseases or disorders in rodents has no scientific justification. In an interview conducted with Dr Samsel, the farmwars.info website reports on the presence of glyphosate in three different rodent diets. The information reported on the website is not supported by sufficient detail or a reference to permit a full scientific review. In conclusion, no new scientific elements were provided that would impact on the validity of regulatory feeding tests in the EU. © European Food Safety Authority, 2015. From (EFSA, 20151006)

4. THIRD PUBLICATION OF SERALINI ET AL.: DEAD COWS OF GLOECKNER Gottfried Gloeckner and Gilles-Eric Séralini claim that Bt-maize has caused the death of some of the cows: (Gloeckner Gottfried & Seralini Gilles-Eric, 20160122)

Gloeckner Gottfried, & Seralini Gilles-Eric. (20160122). Pathology reports on the first cows fed with Bt176 maize (1997–2002) and short comment. Scholarly Journal of Agricultural Science (the domain no more available since January 27), 6(1), pp. 1-8. This domain name expired on Jan 27 2015 06:39PM AND http://www.ask-force.org/web/Seralini/Gloeckner-Seralini-Pathology-Reüprts-Bt176- Maize-Cows-201601.pdf AND short comment http://www.ask-force.org/web/Seralini/Seralini-Experience-first-GM-crop-Europe- short-20160122.pdf

“On an independent modern farm followed by certified veterinarians, dairy cows (mean of 62 per year) were maintained in optimized milk production for 3 years each. From 1997 to 2002, just after the commercial release of the first GMO (genetically modified organism) in Europe, genetically modified (GM) Bt176 maize grown on the farm was progressively introduced in controlled diets. The results are described in the following account, which has an historical value as it is the longest and first on-farm observation of mammals, performed by an experienced farmer and veterinarians, during a period of unusual pathological problems in cows receiving a GMO-rich diet. Thus it was not designed as a scientific experiment. Over the years, and coinciding with regular increases in GMO content of the diet (0–40%), the proportion of healthy cows with high milk yield diminished from 70% (normal rate) to only 40%. At the peak of mortalities in 2002, 10% of the cows died, preceded by a long-lasting paresis syndrome without hypocalcemia or fever, but with kidney biochemical failure and mucosa or epithelial problems. No microbial origin was identified, though intensively investigated. The GM maize, subsequently withdrawn from the market, was at the time the only intended managerial change for the cows. It is proposed that it provoked long-term toxic effects on mammals, which are not observable in most common conditions of intensive farming with high and rapid animal turnover and with no specific labels on GM feed (identifying amount and precise identity of GMO content). More long-term assessments during GMO feeding trials should be performed.” From (Gloeckner Gottfried & Seralini Gilles-Eric, 20160122)

Strangely, the paper published in the “Scholarly Journal of Agricultural Science vanished, since the domain expired on January 27. It is still obtainable over the internet. The journal belongs to the so-called predator journals, which have all sorts of strange habits, as here disappearing from the internet because the domain 30

has been rented only for a limited time. For all further details see (Beall Jeffrey & Scholarly-open-Access, 2013-2015; Bohannon John, 20151119).

Instead of summarizing numerous rebuttals, here the comprehensive review of the Genetic Literacy Project:

Staropoli Nicholas. (20160128).The story behind Séralini’s disappearing GMOs-are-toxic study, and the journal that published it. Genetic Literacy Project,, pp. 4. https://www.geneticliteracyproject.org/2016/01/28/story-behind-seralinis-disappearing-gmos- toxic-study-journal-published/ AND http://www.ask-force.org/web/Seralini/Staropoli-Story-behind-disappearig-Gloeckner-Seralini- study-20160128.pdf

The story behind Séralini’s disappearing GMOs-are-toxic study, and the journal that published it https://www.geneticliteracyproject.org/2016/01/28/story-behind-seralinis-disappearing-gmos-toxic-study-journal-published/

Nicholas Staropoli | January 28, 2016 | Genetic Literacy Project

Where did the latest Séralini anti-GMO paper go?

In a bizarre twist involving the French scientist whose controversial studies are the bedrock of the anti- GMO movement, Gilles-Éric Séralini told Retraction Watch that he has no idea.

Here’s the story.

Gilles-Éric Séralini newest attack on GMOs, this one purporting to show that a type of genetically modified corn once used in livestock feed can be toxic, has disappeared from the web. Séralini and his research group, CRIIGEN — Committee for Independent Research and Information on Genetic Engineering — held a news conference on January 26 announcing:

… New scientific data on Bt-toxins and a thorough study of the records show that this GMO Bt maize is most probably toxic over the long term. 31

Co-authored with Gottfried Glöckner, a German dairy farmer involved in the experimental testing of the corn known at Bt176 (developed by Novartis, now Syngenta) during the 1990s, the article appeared in the Scholarly Journal of Agricultural Sciences (SJAS). Never heard of it? It’s an obscure journal — or was a journal until it disappeared one day after the Séralini paper was published. Based in Nigeria, it was known as an ‘open access’ predatory journal because it its business model was to print controversial, often low quality articles–but only after the authors paid high fees, supposedly to cover editorial services and professional review. They often prey on scientists who are desperate to add publications to their resume, or scientists, like Séralini who cannot get their work published any more in serious, scholarly publications.

Considering Séralini‘s tainted reputation — he has published numerous controversial articles claiming that GMO foods are dangerous that have been widely discredited by regulatory oversight agencies — the mainstream press ignored the event. Two members of the European Parliament spoke out in support of the study. Michèle Rivasi, of France, an environmentalist MEP and member of the Environment and Health Committee in the European Parliament noted the study was proof the precautionary principle should dictate GMO policy. Carey Gillam, the former Reuters reporter renowned for her anti-GMO reporting — after being separated from the global news service, she took a job as “research director” at the anti-GMO organic industry funded U.S. Right to Know, tweeted her admiration for Séralini’s perseverance in informing the public about the ‘dangers’ of GMOs:

The article was gaining some traction in anti-GMO community when the day after it’s release, poof, it disappeared from the web. Readers who click on the link for the article here go to an empty page, with a note saying that the domain expired on January 27.

The anti-biotech website GMWatch, which is run by the same people who oversee Séralini’s propaganda site, GMOSeralini, continues to hype the article. According to GMWatch, the journal also published a commentary by Seralini, which detailed Glöckner’s expertise in this area, but the link to goes to the expired site. This was recovered from way back machine, and posted on Retraction Watch:

When German farmer Gottfried Glöckner began feeding his cows on GM Bt176 maize in 1998, he noticed strange symptoms appearing in the animals. They included partial paralysis (paresis), accompanied by severe fatigue and problems in the kidneys and mucous membranes followed by death in 10% of cases.

Initially the culprit was thought to be a bacterial or viral disease. Analyses were conducted, including some by university laboratories, in collaboration with the German Ministry of Health and the GM maize developer company, Syngenta (then Novartis). But the investigations drew a blank. 32

By 2002 the proportion of GM Bt-maize in the diet, which had been progressively introduced, had reached 40%. Glöckner had become convinced that Bt-maize was the cause of the diseases in his cows. He sued Syngenta and was awarded partial compensation for his losses. But he subsequently suffered legal and personal setbacks, as reported in a commentary newly published by Prof Gilles-Eric Séralini in the peer- reviewed Scholarly Journal of Agricultural Sciences (SJAS) (Séralini GE, The experience of one of the first GM crop farmers in Europe, SJAS 2016).

After the court cases ended, Prof Séralini gained access to the veterinary records and to detailed archived data for each cow, as well as to the testimony of the farmer, who holds a Master’s degree in Agricultural Sciences.

Analysing Gottfried Glöckner’s story

In the 1990’s, Gottfried Glöckner and his wife owned a farm in Weidenhof, Wölfersheim, Hessen, Germany. The couple grew conventional crops like corn and raised livestock. When GMOs began to be available for farmers in the mid 1990’s, Glöckner was initially very intrigued by this new technology. He told The Ecologist in 2014:

Since 1995 when genetically modified Roundup-Ready (RR) Soy was imported into the EU, approved under the principle of ‘substantial equivalence,’ I had been interested in the subject GMO technology in plants. When in 1997 the EU approved the commercial sale of Syngenta GMO corn (Syngenta Bt176) I decided, as a farmer interested in new better technologies, to grow Syngenta’s Bt176 on my land.

By 2002, 40 percent of the feed he was giving his cows was derived from Syngenta’s Bt176 maize — a GMO corn that possesses the gene for a bacterial toxin that is also an organic Bt-insecticide. Yet around this time, Glöckner started to become worried about the safety of these crops. In 2001, a few of his cows became lethargic, weak, and had reduced milk yield. That year, five of his 66 cows died. A similar scenario played out in 2002 and seven dead cows. After conducting his own investigation, Glöckner concluded that the culprit for the death of his cows was the Bt-corn. The incident turned into a large news event in Germany and regulatory agencies were called to investigate.

In the 15 years since the incident, Glöckner fought a long legal battle with Syngenta and was incarcerated for violence against his wife. Now that the legal battles are over the data and veterinarian record were finally released and Séralini was able to get a hold of it and decided to publish the data.

How did these cows die?

Many critics of the paper have pointed out the confusing nature of the opening statement of the paper in which the authors admit, “This study was not designed as a scientific experiment.” The phrase is also repeated in the paper’s abstract. Yet despite this self-characterization as not science, it has been promoted as a scientific evidence against the safety of GMOs.

Yet, putting aside some of the quirks of this paper and the journal, there is still a legitimate question of what happened to these cows. Prof. Dr. Klaus-Dieter Jany, who was the head of the Molecular Biology Centre of the Federal Research Centre for Nutrition and Food in Germany from 1998-2008 and was asked at the time by the Ministry of Agriculture for an assessment, investigated the farm and the cows shortly after the tragedy. 33

In his expert opinion, the evidence for the cause of the cow’s deaths was most likely attributed to one of a few factors which all revolve around Glöckner’s use of inappropriate feeding practices. Jany says the farm was using, “inappropriate power feed and a wrong feed composition to increase the milk yield in a short time.” Furthermore, he notes that they found Glöckner was using, “inadequate and unsanitary corn silage with many undesired contaminations (plastics and dead rats) of the corn silage.” Finally, Jany says botulism also couldn’t be ruled out as at least a potential cause for at least some of the deaths.

Jany’s explanation is also consistent with the vast majority of research into the effects of GMOs in the feed of animals. Most notably, a large one from 2015 published in Journal of Animal Science which used data from 100 billion animals and found no health dangers to the animals from GMOs. But large data sets might not be needed to conclude that GMOs in animal feed are safe. Today, almost all of livestock feed today is derived from GMOs, and if they weren’t safe to consume Séralini wouldn’t need to go looking for the one farmer who lost 12cows.

Séralini had apparently hoped that this new “study” would stir the embers of concern around GMO food safety. The CRIIGEN news release:

This study reveals once again the urgent need for specific labelling of the identity and quantity of GMOs, especially in food and feed. Long-term testing of GM food and the pesticides they are designed to contain must be carried out and made public. This is now more essential than ever.

In fact, this study reveals only one thing: it’s critical that research on biotechnology, food and farming practices be carried out by credible researchers publishing in top tier journals reviewed by respected independent international science agencies. That leaves Séralini on the side-lines.

Nicholas Staropoli is the Associate Director of GLP and Director of the Epigenetics Literacy Project. He has an M.A. in biology from DePaul University and a B.S. in biomedical sciences from Marist College. Follow him on twitter @NickfrmBoston.

Gottfried Glöckner fed his cows with a maize breed (Bt-176) which has been withdrawn, the Bt-toxin lever was considerably higher than the follow-up Bt-maize traits. This is also the reason, why multiple feeding studies have been conducted, all with negative results, no measurable toxicity, here just one example from

Petit, L., Baraige, F., Bertheau, Y., Brunschwig, P., Diolez, A., Duhem, K., Duplan, M. N., Fach, P., Kobilinsky, A., Lamart, S., Schattner, A., & Martin, P. (2005). Detection of genetically modified corn (Bt176) in spiked cow blood samples by polymerase chain reaction and immunoassay methods. Journal of Aoac International, 88(2), pp. 654-664. ://000228000600036 AND http://www.ask-force.org/web/Bt/Petit- Detection-Bt-Cowblood-2005.pdf

The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1 Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35s promoter and Btl76specific junction sequence) was determined by both the polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) and the 5’-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26s rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (asl-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/ml in 10 mL whole blood samples. The sandwich ELlSA kit used detected down to 1 ng transgenic Cry1 Ab protein/ml spiked whole blood. From (Petit et al., 2005)

The same harmless results have been published by many authors: (Aeschbacher et al., 2005; Badosa et al., 2004; Barriere et al., 2001; Bordoni et al., 2004; Brake & Vlachos, 1998; Brusetti et al., 2004; Chiueh et al., 2004; Corbisier et al., 2007; EFSA-Opinion, 2006; El-Sanhoty et al., 2006; Felke et al., 2010; Feriotto et al., 2003; Ferrini et al., 2007; Garcia-Canas et al., 2004; Gardenghi et al., 2004; Lutz et al., 2006; Petit et al., 34

2005; Ramadan et al., 2006; Siegfried et al., 2001; Taverniers et al., 2005; Tony et al., 2003; Tony et al., 2004; Trabalza-Marinucci et al., 2008; Verdenius, 1990; Visser et al., 1994; Zangerl et al., 2001; Zoerb et al., 2003).

It is hardly necessary to say that in the Glöckner-Seralini study only two of the above studies were cited and shortly commented. And also the following two modern studies with the ultimate proof of Bt-toxins being harmless are also not mentioned, although this would be closer to reality, since the Bt-176 production and distribution has long since been stopped, and the only negative incidence known today are the 12 dead cows of Gottfried Glöckner, where studies and results of independent experts have ruled out the possibility of Bt-167 toxins, see the summary of the expertise, done at the time of the incidences in Glöckner’s stable:

5. CAVEAT ABOUT PSEUDO-BALANCE OF VIEWS IN THE GM DEBATE

A caveat at the end of this paragraph on peer review is appropriate: Although it is in principle necessary to ask ethical questions, we should first concentrate on the scientific assessment of a professional peer review strictly following a factual agenda such as (Chassy, B. & Parrott, 2009; Chassy, Bruce M., 2009) are demanding. Only then, when this filter has been passed successfully, then it is important to go into ethical and socio-economic questions. But as often, it is the farmers and the market regulating efficiently – and – no surprise – they follow quite naturally socio-economic principles. It is wrong to mix scientific and ethical questions as de Melo et al. and Inteman et al. are asking for (de Melo-Martin & Meghani, 2008; Intemann & de Melo-Martin, 2008), the result is then to erroneously accept for discussion a paper like the one of Seralini et al. (Seralini, G.E. et al., 2007), a paper which has been seriously and repetitiously criticized on a factual basis by EFSA (EFSA- Statement, 2007; EFSA, 2007a, 2007b). Such papers should not be seen as a publication which takes into account a “balanced view”, because they are flawed in the first place. Papers from the laboratory of Séralini are unfortunately often cited as done by independent scientists with the function of important wistle- blowers, which is not very convincing: Digging into the financial support of Séralini and his CRIIGEN lab is highly interesting, You realize that they also receive funds which come from opponents of GMO technology, such as Sevene Pharma, commercializing homeopathic products which claim to detoxify various toxic products (Imposteurs, 2011) and more: CRIIGEN has been created with the financial support of the retailer Carrefour which has also contributed financially to certain studies of Séralini and his group. Interestingly enough, Carrefour, the second largest food distributor in the world, sells its own brand of “GMO-free” products… Source: (Kuntz, 2011). For more details about ethical questions see chapter 8.6. But it should also be seen here that the main argument of the quality of scientific papers is not who actually financed the research, but one should scrutinize the methodological and scientific quality of the papers. It is rather simplistic to split the science paper world into two half’s with a contrasting financial background. It is not only ironic, but truly sad that a colleague of ours, Marc Fellous, has been convicted by a French court to have accused Gilles Eric Séralini for not being a neutral scientist anymore (taking money from Greenpeace, which is a fact). Interestingly enough Fellous has not been convicted by the same court because he is accusing Séralini for publishing bad science…(Kuntz, 2011; Olenyi Sebastian, 20120228; Sachs, 2005; Sachs et al., 2009, 20070624). See also more material about the case Seralini-Fellous in (Seppi Wackes et al., 2011). 35

An interesting paper concentrates on risk perception, a wider view in toxicology from scientists of all kinds and working environment to lay people, it may help to arrive to differentiated insights, as developed by Nancy Kraus (Kraus et al., 1992).

“Human beings have always been intuitive toxicologists, relying on their senses of sight, taste, and smell to detect harmful or unsafe food, water, and air. As we have come to recognize that our senses are not adequate to assess the dangers inherent in exposure to a chemical substance, we have created the sciences of toxicology and risk assessment to perform this function. Yet despite this great effort to overcome the limitations of intuitive toxicology, it has become evident that even our best scientific methods still depend heavily on extrapolations and judgments in order to infer human health risks from animal data. Many observers have acknowledged the inherent subjectivity in the assessment of chemical risks and have indicated a need to examine the subjective or intuitive elements of expert and lay risk judgments. We have begun such an examination by surveying members of the Society of Toxicology and the lay public about basic toxicological concepts, assumptions, and interpretations. Our results demonstrate large differences between toxicologists and laypeople, as well as differences between toxicologists working in industry, academia, and government. In addition, we find that toxicologists are sharply divided in their opinions about the ability to predict a chemical's effect on human health on the basis of animal studies. We argue that these results place the problems of risk communication in a new light. Although the survey identifies misconceptions that experts should clarify for the public, it also suggests that controversies over chemical risks may be fueled as much by limitations of the science of risk assessment and disagreements among experts as by public misconceptions.” (Kraus et al., 1992)

6. Final Word About Seralini And Criigen Activities, An Analysis From Jon Entine

A full copy of an article from the Genetic Literacy Project, it contains a retrospective of all the disputes around the work of Gilles Eric Séralini from Jon Entine (Entine Jon, 20130610)

Entine Jon. (20130610). Being Gilles-Eric Seralini: Inside the mind of the anti-GM movement. Genetic Literacy Project,, pp. 6. https://www.geneticliteracyproject.org/2013/06/10/being-gilles-eric-seralini-inside-the-mind-of-the-anti-gm-movement/ AND http://www.ask-force.org/web/Seralini/Entine-Being-Gilles-Eric-Seralini-Inside-mind-20130610.pdf

Being Gilles-Eric Seralini: Inside the mind of the anti-GM movement

Jon Entine | June 10, 2013 | Genetic Literacy Project https://www.geneticliteracyproject.org/2013/06/10/being-gilles-eric-seralini-inside-the-mind-of-the-anti-gm-movement/

An anti-GMO brain. (Created from image via Flickr/Ethan Hein)

In one of the weirdest scenes in Being John Malkovich, the endearingly offbeat 1999 comedy-fantasy that takes us into mind of actor John Malkovich playing a fictional version of himself, Malkovich’s doppelgänger enters his own subconscious and is placed in a world where everyone looks like him and can only say “Malkovich.”

This scene is equally disconcerting and compelling because it provides a metaphor for being trapped in your own narrow view of the world—it’s just you, you, you. 36

This is what appears to be going on with anti-crop biotechnology campaigners. All they can see and hear are their own slogans and conspiracy theories—it’s just Frankenfood, Frankenfood, Frankenfood.

The June 4th Washington, DC crop biotech forum—which you can view in full on the Genetic Literacy Project website—was supposed to have been a debate featuring two of the world’s most prominent anti-biotech activists—Gilles-Eric Seralini, a French scientist, and Jeffrey Smith, who runs the Institute for Responsible Technology, an anti-biotech NGO. Both ran tail, apparently when they realized this event would require more than talking to themselves—they’d actually have to engage with mainstream science. Malkovich, at least, walked the edge of self-mockery and is shamed by his subconscious self-centeredness. That’s not the case with Seralini and Smith.

As I began assembling this event, two leading anti-biotechnology groups—the Center for Food Safety and the Union of Concerned Scientists—turned down my invitation. I don’t believe the “experts” at either organization represent mainstream scientific thinking—but they are the best the Antis have. Disgracefully, they would not engage in open dialogue.

As I’ve written before, in controversies you don’t get to choose your opponents — the public does that for you. That’s what led me to Smith and Seralini. Smith, while not a scientist — he is a former yogic instructor and political candidate—has written two best- selling self-published books and produced a documentary narrated by the wife of Dr. Mehmet Oz, whose show he has appeared on numerous times. Whatever his credentials, he’s a favourite of Hollywood, and one of the most quoted anti-biotech crusaders in the world.

Gilles-Eric Seralini. (CREDIT: Flickr/ALDEADLE Alliance of Liberals and Democrats for EU)

To facilitate dialogue, Smith and I agreed to invite professor Seralini, whose paper released last fall claiming that rats fed genetically modified corn developed cancerous tumors turned him into a global celebrity. The paper was crudely ideological and roundly rejected by established scientists and every major science and science journalism organization of note, particularly experts in Europe where the public remains very leery of biotechnology. Seralini knows this: he has denied every request to share his data with independent researchers. That’s why I was surprised when he agreed to participate… but not surprised when he, along with Smith, backed out once I recruited Kevin Folta, Interim Chair of the Horticultural Sciences Department at the University of Florida, to join the debate.

At the June 4th forum, Kevin would ultimately deconstruct both Smith’s and Seralini’s bizarre takes on crop biotechnology. Facing the prospect of a civil but rigorous debate, Smith and Seralini scurried for the exits.

After they pulled out, Kevin and I retooled the debate into a forum and recruited Karl Haro von Mogel, University of Wisconsin plant breeding and plant genetics PhD candidate, plant biotechnologist, and cofounder of the Biofortified website.

Reason, religion and food security

It’s tempting to characterize this dispute as a battle between science and ideology, but that would be simplistic. Issues of risk, notions of “Nature” and “natural”—these are emotional, even religious beliefs. How we view food is deeply personal. Science can only take us so far—but it’s pretty far.

We all can appreciate why any debate over farming, food and modern technology tends toward contentious. After all, we are talking about our children and our health. The one thing we can agree on is that everyone, on all sides of this discussion, wants abundant, highly nutritious food produced with the least environmental damage.

We decided to focus the forum on global food security. According to the Food and Agriculture Organization (FAO) of the United Nations, 868 million people are undernourished. Food security has improved in recent decades, as the undernourishment rate dropped from 18.6% in 1990-1992 to 12.5%. But over the same period, population grew from 5.4 to 7.0 billion—and it’s on track to grow to 9 million over the next two to three decades. As Bill Gates once said, “The world is getting better, but it’s not getting better fast enough, and it’s not getting better for everyone.” 37

Global food security is a complex challenge. Agriculturally rich regions like North America, Argentina and Brazil must produce enough to make up for production deficits in Asia, Africa and even Europe. We will need 70-100% more food by 2050 to match population and prosperity growth, and it must occur in the face of more frequent extreme weather events marked by floods, droughts and heat waves.

One thing we know is that technology—including biotechnology—must play a central role, just as it did in the Green Revolution. Beginning in the late 1940s, genetic research led to the breeding of high-yield grains. Combined with the use of new fertilizers, herbicides and pesticides, output soared. Since 1950, world wheat production alone has increased by more than 300%.

That said, the environmental consequences of high-yield industrial scale farming are daunting. Organic farming can play a role, but it’s at best a marginal part of the solution. Although no one believes it’s a silver bullet, the overwhelming consensus of the science and farming communities is that genetic engineering can and will be part of farmers suite of tools for addressing increasing food needs while mitigating environmental damage.

A just released research paper in the Public Library of Science open access journal, PLoS One on genetically modified Bt-cotton says it best:

Controlling for other factors, the adoption of GM cotton has significantly improved calorie consumption and dietary quality, resulting from increased family incomes. This technology has reduced food insecurity by 15–20% among cotton-producing households. GM crops alone will not solve the hunger problem, but they can be an important component in a broader food security strategy.

What is agricultural genetic engineering?

The Food and Drug Administration recently addressed its role in regulating food safety and the relative riskiness of GE crops. Selective breeding turned inedible wild grains, like corn and wheat into delicious modern varieties. We’ve been doing it for thousands of years, but it is terribly imprecise.

As the FDA notes, “These genes [from traditional selective breeding] may include the gene responsible for the desired characteristic, as well as genes responsible for unwanted characteristics.” Genetic engineering, rather, can “introduce the desired characteristic without also introducing genes responsible for unwanted characteristics.”

Introduced first in the US in the mid-1990s, genetically engineered crops are now grown by more than 17 million farmers in 28 countries. Eighty-one percent of the world’s soybeans, 81% of cottonseed, 35% of corn and 30% of canola are now grown using GE seeds. In 2012, the global area of biotech crops continued to increase for the 17th year. Their commercial value worldwide exceeds $185 billion per year. In the US, the use of herbicide tolerant soybeans, cotton and corn, and pesticide-resistant Bt-cotton and corn is even more pronounced.

Almost all GE crops are based on two well-established and rigorously tested technologies. First, Bt-crops produce a bacterial protein known as Bacillus thuringiensis. It’s naturally occurring—and it’s widely used by organic farmers to selectively kill pest insects. Genetically engineered Bt crops simply produce their own Bt. The effects are identical to what happens on organic farms— which is what makes protests against genetically engineered Bt crops seem so bizarre to scientists. The net result is that Bt-crops increase yields because farmers lose fewer crops to insect pests.

The other major GE crops are those designed to be herbicide-resistant, most commonly glyphosate, better known as Monsanto’s Roundup. Glyphosate does not bio-accumulate and breaks down rapidly in the environment. Because the weed killer is more powerful and less toxic than the chemicals that it competed with, farmers quickly adopted glyphosate—even more so after Monsanto introduced genetically engineered versions that were paired with the herbicide. The use of crops engineered for herbicide resistance reduces inputs, cost, and labour for farmers. It’s agricultural sustainability at work.

In the US, the use of herbicide tolerant soybeans, cotton and corn, and pesticide resistant Bt-cotton and corn has soared since their introduction. It’s estimated that 90% of the farmers around the world that grow GE crops—roughly 15 million people—are resource-poor. The total acreage of GE crops in developing countries now exceeds that of the developed world.

Despite the worldwide boom, the controversy has not abated, particularly in Europe, where GE crops for food are banned or heavily restricted. Many people, particularly those who call themselves progressives, are convinced that genetically modified foods are either harmful or have unknown medical and ecological impacts. These views are promulgated on anti-biotech sites like Natural News.com, Mother Jones and the Organic Consumers Association—but they even infect normally science-based sources like the New York Times.

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Pulitzer journalist Moss embarrasses New York Times

The latest embarrassment came two weeks ago, when Pulitzer Prize winning reporter Michael Moss appeared in the paper’s video cast discussing genetically modified crops. He framed the debate in apocalyptic terms. Here is an excerpt:

I have family in Europe. They’ve been talking to me about GMOs for years and years. I think they decided that even though there is no hard science showing long-term health problems with GMOs, they also point out that the research really hasn’t been done. So for them the glass is half empty, rather than half full. They’re saying, “look until proven safe, we’re gonna, like, avoid this stuff.”

I think it’s been under the radar a bit. In increasing mood, people are concerned about it. Those [anti-Monsanto] rallies over the weekend were amazing. So many people hit the streets and I think part of the thing happening here is people are realizing, this is really scary stuff. I mean, just consider the name, right. Genetically modified organisms. This isn’t like taking one apple and crossing it with another and getting a redder, shinier apple. This is extracting genetic material from one living creature and putting it another. And that’s really disturbing to people.

There are so many inanities it’s difficult to know where to begin. Particularly appalling was his “really scary stuff” comment—it is fear mongering promoted by the world’s most respected newspaper.

Moss was flat out wrong when he said: “research [on genetically modified foods] really hasn’t been done.” There have been 1,000 or more studies on GM crops and foods. As von Mogel outlined in his talk at the biotech forum, more than 600 hundred of them are catalogued (and soon to be searchable) at the Biofortified site’s GENERA database.

Couldn’t Moss have talked to scientists instead of relying on paranoid gossip from family in Europe? For an investigative reporter, this seems a startling lapse. There is broad scientific consensus that genetically engineered crops currently on the market are safe to eat.

Here is what some of the world’s major science groups have publicly declared about crop biotechnology.

 The American Academy for the Advancement of Science (AAAS): “The science is quite clear: crop improvement by the modern molecular techniques of biotechnology is safe.”

 The U.S. National Academy of Sciences: “[N]o adverse health effects attributed to genetic engineering have been documented in the human population.”

 The World Health Organization: “No effects on human health have been shown as a result of the consumption of such foods by the general population in the countries where they have been approved.”

 The European Commission: “[N]o scientific evidence associating GMOs with higher risks for the environment or for food and feed safety than conventional plants and organisms.”

 The Union of the German Academies of Science and Humanities, Germany’s most prominent science organization in the heart of anti-biotech Europe: “Foods from approved GM crops are safe for humans and animals; Approved GM crops do not pose environmental hazards.”

All three forum participants—me, von Mogel and Folta—addressed the conspiracy theory ruminations of many anti-biotech campaigners. For instance, the belief that genetic engineering is an abomination, a scar against the “natural order”, which they believe is good and safe. This is the “naturalistic” fallacy, an erroneous simplification of nature’s ecological impact.

Activists often paint a scary picture of modern technology: Mad scientists holed up in their labs tinkering with the keys to life itself, playing god. Genetic engineering is portrayed as untested and risky—; something is bound to go horribly wrong. To judge from their rhetoric, what they call the “Frankenfood revolution” courts health epidemics and environmental Armageddon. That’s the kind of language used by tens of millions of people—painting apocalyptic scenarios of a Brave New World.

The activists’ greatest ire is focused on Monsanto. It is the evil face of a scary industry—the spawn of the Devil, in their eyes. Being first out of the box with a new technology created an inviting target. Combined with the massive popular mistrust of government, we have a recipe for generating anti-globalism nightmares. Monsanto, activists say, is trying to take over the worlds’ food supply and government regulators are its willing puppets.

The NGO and fringe media obsession with conspiracy theories threatens to obscure pioneering developments in crop biotechnology—what could be called Biotech 2.0—which more clearly benefit consumers and the environment. That was the focus of von Mogel’s talk. He outlined more than a dozen new products, including:

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 Crops that eliminate or greatly reduce the need for nitrogen fertilizers  Drought and flood resistant crops of all varieties and grains that can grow in saltier water and soils  Carbon sequestering trees to address global warming  Nutritionally enhanced grains and vegetables that produce healthful omega-3 fatty acids  Salmon that grows twice as fast as conventional varieties and consumes fewer resources—it’s more sustainable—and is just as nutritious  Fruits and nuts with allergen-causing proteins knocked out; imagine hypo-allergenic peanut butter These innovations have been tested and found safe, but languish on laboratory shelves, victims of activist campaigns that paralyze the approval process in country after country.

Whatever one thinks of their credentials, Smith and Seralini have emerged as potent voices, their views echoed on thousands of ‘natural’ or ‘organic’ or progressive media sites. Yet they both promote blatant falsehoods.

What’s the take away from the debate that wasn’t? Smith, Seralini and their adherents, like the fictional John Malkovich, are trapped in their own minds—unable or unwilling to embrace critical thinking or rely on empirically based evidence. Worse, well- meaning people who should know better, like Michael Moss, spread these fear. They’ve contributed to what British journalist Mark Lynas has characterized as one of the greatest science communication disasters of the past half-century—the dangerous misinformation campaign over genetically engineered crops and foods.

Jon Entine, executive director of the Genetic Literacy Project, is a senior fellow at the Center for Health & Risk Communication and STATS (Statistical Assessment Service) at George Mason University. Much more stories about Séralini see on the website of the Genetic Literacy Project, and type the keyword Seralini: you will find a total of 8 pages of references, all related to the Seralini debate: https://www.geneticliteracyproject.org/page/6/?s=Seralini&submit_x=5&submit_y=4#038;submit_x=5&submit_y=1

7. TWO NEW SUMMARIES OF SOME WORST PSEUDOSCIENCE PUBLICATIONS UNDERPINNED WITH STATISTICS

Panchin, A. Y. (2013). Toxicity of roundup-tolerant genetically modified maize is not supported by statistical tests. Food and Chemical Toxicology, 53(0), pp. 475. http://www.sciencedirect.com/science/article/pii/S0278691512007843 AND http://www.ask- force.org/web/Seralini/Panchin-Toxicity-roundup-tolerant-GM-maize-not-supported-by-statstics-2013.pdf

Panchin, A. Y., & Tuzhikov, A. I. (2016). Published GMO studies find no evidence of harm when corrected for multiple comparisons. Critical Reviews in Biotechnology, pp. 1-5. http://dx.doi.org/10.3109/07388551.2015.1130684 AND http://www.ask- force.org/web/Seralini/Panchin-published-GMO-studies-no-evidence-harm-2016.pdf

“A number of widely debated research articles claiming possible technology-related health concerns have influenced the public opinion on genetically modified food safety. We performed a statistical reanalysis and review of experimental data presented in some of these studies and found that quite often in contradiction with the authors’ conclusions the data actually provides weak evidence of harm that cannot be differentiated from chance. In our opinion the problem of statistically unaccounted multiple comparisons has led to some of the most cited anti-genetically modified organism health claims in history. We hope this analysis puts the original results of these studies into proper context.” From (Panchin & Tuzhikov, 2016)

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The following paragraphs are an excerpt from (Panchin & Tuzhikov, 2016), it is the statistical analysis and comments on the papers of Séralini:

Next, we will review the study by Seralini et al. (Séralini et al., 2012, 2014). The authors chose to drop the ‘‘statistics part’’, related to the main conclusions of the article, all together [Panchin, 2013), 21] so we had to obtain the p values ourselves using a straight-forward and transparent statistical significance test used in the analysis of contingency tables – Fishers exact test (other tests would yield similar results). In the current reanalysis of the hypothesis presented by Seralini et al., the numerical data was taken from the original article. The number of multiple comparisons is discussed but is not relevant in this case because the study reveals no significant effects of GM food on animal health even without the Bonfferoni correction. In this study, male and female rats were fed genetically modified maize (GMM) in three different concentrations (11%, 22% and 33%) with and without an added herbicide (six GMM-fed groups for each sex). A modest control set of rats (N 1/ 4 10 for each sex) was used. One of the main conclusions was that ’’in females, all the treated groups died 2–3 times more than controls’’. As follows from the article: 2 female rats out of 10 died before the mean survival time in the control group, compared to 29 out of 60 in the pooled six GMM- fed groups. However, this difference is not statistically significant (p 1/ 4 0.09, two-tailed Fishers exact test, not adjusted for multiple comparisons).

Seralini et al. state: ‘‘20% females (only two) died spontaneously, while up to 50% males and 70% females died in some groups on diets containing the GMM’’. The highest mortality was observed for the group of female rats fed 22% GMO: 7 out of 10 rats died prematurely. Even if we compare the mortality in this particular group of rats with controls, we conclude that the observed difference is not statistically significant (p 1/ 4 0.07, two-tailed Fishers exact test, not adjusted for 12 multiple comparisons). In addition, mortality rates among female rats were lower in the 33% GMO group comparing to the 22% GMO group, thus a dose– response effect was not observed.

Among males, 3 rats out of 10 died prematurely in the control group, compared to 19 out of 60 rats in the six pooled GM-fed groups (not significant p 1/ 4 0.615, two-tailed Fishers exact test). One of the most curious facts is that if we set aside statistical significance, take the data provided by Seralini et al. and use same ‘‘cherry picking’’ strategy, we could state that ‘‘in males, groups with 22% and 33% GMM in their diet died 3 times less than controls’’ (p 1/ 4 0.291, two-tailed Fishers exact test, not significant). Does a high dose of GMM prolong the lifespan of male rats? The authors did not report this ‘‘observation’’. The article states: ‘‘in treated males, liver congestions and necrosis were 2.5–5.5 times higher’’ and that ‘‘females developed large mammary tumors almost always more often than and before controls’’. Two male rats out of 10 had liver pathologies in the control group, compared to 30 out of 60 GMM fed male rats. About 5 female rats out of 10 developed 41

mammary tumors in the control group, compared to 44 out of 60 GMM fed female rats. Neither of these differences are statistically significant (p 1/4 0.076 and p 1/4 0.133, two-tailed Fishers exact test). 4 A. Y. Panchin et al. Crit Rev Biotechnol, Early Online: 1–5 Downloaded by 5.228.65.227] at 09:09 15 January 2016 Over 30 different organs were analyzed in the study and the two sexes were analyzed independently. Seralini et al. provide the data selectively, however none of the reported differences between the number of rats with specific organ pathologies in control and GMM fed groups are statistically significant even at alpha 1/4 0.05 (two-tailed Fishers exact test without the Bonferroni correction). In his response to the criticism of the absent statistical analysis, Dr. Seralini gave a laconic reply: ‘‘Statistics do not tell the truth, but may help in understanding results’’. [22] Indeed and clearly so! As a side note: the rat tumors presented as scary imagery in the study are frequent in Sprague– Dawley rats and have a reported spontaneous incidence of 45% over a 1.5-year period.[23

8. BIBLIOGRAPHIES RELATED TO SERALINI THREE MAJOR PAPERS

Comprehensive overviews on GM food regulation has been published by (Kuiper & Kleter, 2003) and a summary of the politically motivated disputes is given by (Sinha, 2009)

A Bibliography related to the Séralini long term study: (Ammann Klaus, 20121111)

Thematic bibliography long-term-experiments, Séralini and News from Web of Science and other sources, with full text links Version January 10 .2013 pp 63 (Manuscript) http://www.ask-force.org/web/Seralini/Ammann-Thematic-Bibliography-Longterm-Seralini-20121111.pdf

ABNE. (2012). FOLLOW-UP ON CONTROVERSIAL GM MAIZE STUDY BY SERALINI et al; European Food Safety Authority confirms their conclusion that the study is not valid for risk assessment pp. Ouagadougou, Burkina Faso ISBN/ISSN African Biosafety Network of Expertise (ABNE) Retrieved from http://www.nepadbiosafety.net/abne/wp-content/uploads/2012/12/Follow_up_NK603-Maize-Study.pdf AND http://www.ask- force.org/web/Seralini/ABNE-Followup-Seralini-2012.pdf

Academies Francaises. (20121018-19). Avis des Académies nationales d’Agriculture, de Médecine, de Pharmacie, des Sciences, des Technologies, et Vétérinaire sur la publication récente de G.E. Séralini et al. sur la toxicité d’un OGM pp. 5 Paris, France ISBN/ISSN Academies d'Agriculture de France, Académie Nationale de Médicine, Académie Nationale de Pharmacie, Institut de France Académie des Sciences, Académie des Téchnologies, Académie Vétérinaire de France Retrieved from http://www.academie-sciences.fr/activite/rapport/avis1012.pdf AND http://www.ask- force.org/web/Seralini/Academies-Nationales-Avis-Seralini-20121019.pdf AND http://www.ask-force.org/web/Seralini/Academies-Nationales-Avis- Seralini-details20121018.pdf AND http://www.ask-force.org/web/Seralini/Academies-Communique-de-Prssse-Avis-20121019pdf.pdf

Aeschbacher, K., Messikommer, R., Meile, L., & Wenk, C. (2005). Bt176 corn in poultry nutrition: Physiological characteristics and fate of recombinant plant DNA in chickens. Poultry Science, 84(3), pp. 385-394. ://000227113500005 AND http://www.ask- force.org/web/Bt/Aeschbacher-Poultry-Bt176corn-2005.pdf

AJSPI, & Huet Sylvestre. (20121015). Séralini et les OGM: une déclaration de l'AJSPI ( L'Association des journalistes scientifiques ). Liberation. Retrieved from http://sciences.blogs.liberation.fr/home/2012/10/s%C3%A9ralini-et-les-ogm-une- d%C3%A9claration-de-lajspi.html AND http://www.ask-force.org/web/Seralini/AJSPI-Seralini-OGM- Declaration-20121015.pdf

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Ammann, K. (20120807). Review: The Austrian experiment with mice fed with a hybrid GM maize from Monsanto, Part 3.5a: Background and Part 3.5b: Experiment, ASK-FORCE AF-5. ASK-FORCE-5, AF-5, pp. 34. http://www.ask-force.org/web/Debate-MS-Ch/Debate-Ch-3.5-Austrian-Mice- Background-Experiment-20120807.pdf

Ammann Klaus. (20121111, January 10, 2013). Thematic bibliography longterm experiments, Séralini and News from Web of Science and other sources, with full text links Version February 9 .2013. Bibliography. Neuchatel, Switzerland. http://www.ask-force.org/web/Seralini/Ammann- Thematic-Bibliography-Longterm-Seralini-20121111.pdf

Anonymous-Rat-List. (2003). Scientific Literature Survey: Mammary tumors, pituitary tumors, and spaying. 2012, from http://www.ask- force.org/web/Seralini/Anonymous-Rat-List-Spaying-2003.pdf

Aris, A. (2012). Response to "Food Standards Australia New Zealand's" comments. Reproductive Toxicology, 33(3), pp. 403-404. ://WOS:000303361200017 AND http://www.ask-force.org/web/HerbizideTol/Aris-Response-Food-Standards-NZ-2012.pdf

Arjó, G., Portero, M., Piñol, C., Viñas, J., Matias-Guiu, X., Capell, T., Bartholomaeus, A., Parrott, W., & Christou, P. (2013). Plurality of opinion, scientific discourse and pseudoscience: an in depth analysis of the Séralini et al. study claiming that Roundup™ Ready corn or the herbicide Roundup™ cause cancer in rats. Transgenic Research, pp. 1-13. http://dx.doi.org/10.1007/s11248-013-9692-9 AND http://www.ask- force.org/web/Seralini/Arjo-Plurality-Opinion-Scientific-Discourse-Seralini-2013.pdf AND http://www.ask-force.org/web/Facultyof1000/Arjo- Plurality-Opinion-Seralini-F1000-Ammann-20130311.pdf

Badosa, E., Moreno, C., & Montesinos, E. (2004). Lack of detection of ampicillin resistance gene transfer from Bt176 transgenic corn to culturable bacteria under field conditions. Fems Microbiology Ecology, 48(2), pp. 169-178. ://WOS:000221441000006

Bales, J. M., Powell, D. S., Bethel, L. M., Reed, D. S., & Hartman, A. L. (2012). Choice of inbred rat strain impacts lethality and disease course after respiratory infection with Rift Valley Fever virus. Frontiers in Cellular and Infection Microbiology, 2, pp. http://www.frontiersin.org/Journal/Abstract.aspx?s=149&name=cellular_and_infection_microbiology&ART_DOI=10.3389/fcimb.2012.00105 AND http://www.ask-force.org/web/Seralini/Bales-Choice-Inbred-Rat-Strain-2012.pdf

Barale-Thomas, E. (2013). Letter to the editor. Food and Chemical Toxicology, 53(0), pp. 473-474. http://dx.doi.org/10.1016/j.fct.2012.10.041 AND http://www.sciencedirect.com/science/article/pii/S0278691512007867 AND http://www.ask-force.org/web/Seralini/Barale-Thomas-SEPT-Letter- Editor-Seralini-20131116.pdf

Barriere, Y., Verite, R., Brunschwig, P., Surault, F., & Emile, J. C. (2001). Feeding value of corn silage estimated with sheep and dairy cows is not altered by genetic incorporation of Bt176 resistance to Ostrinia nubilalis. Journal of Dairy Science, 84(8), pp. 1863-1871. ://WOS:000170470700014 AND http://www.ask-force.org/web/Bt1/Barriere-Feeding-Value-Corn-Silage-Estimated-Sheep-Dairy-Cows-2001.pdf

Beall Jeffrey, & Scholarly-open-Access. (2013-2015). OMICS Goes from “Predatory Publishing” to “Predatory Meetings”. pp. http://scholarlyoa.com/2013/01/25/omics-predatory-meetings/ AND http://www.ask-force.org/web/Peer-Review/Beall-Scholarly-Open-Access- OMICS-Goes-from-Predatory-Publishing-to-Meetings-2015.pdf

Belgian Biosafety-Advisory. (20121019). Advice of the Belgian Biosafety Advisory Council of the article by Seralini et al. 2012 on toxicity of GM maize NK603 pp. 10 Brussels ISBN/ISSN Belgian Biosafety Advisory Council Retrieved from http://www.bio-raad.be/docs/BAC_2012_0898_CONSOLIDE.pdf AND http://www.ask-force.org/web/Seralini/Advice-Belgian-Biosafety-Advisory-Council-Seralini-20121019.pdf

Benachour, N., & Seralini, G. E. (2009). Glyphosate Formulations Induce Apoptosis and Necrosis in Human Umbilical, Embryonic, and Placental Cells. Chemical Research in Toxicology, 22(1), pp. 97-105. ://WOS:000262602900015 AND http://www.ask- force.org/web/Seralini/Benachour-Glyphosate-Formulations-2009.pdf

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Berry, C. (20121107). Letter to the editor. Food and Chemical Toxicology(0), pp. http://www.sciencedirect.com/science/article/pii/S0278691512007983 AND http://www.ask-force.org/web/Seralini/Berry-Letter-Editor-FCT- 20111107.pdf

BfR-Bund. (20120928). Bundesinstitut für Risikobewertung: Veröffentlichung von Seralini et al. zu einer Fütterungsstudie an Ratten mit gentechnischverändertem Mais NK603 sowie einer glyphosathaltigen Formulie-rung Berlin, Germany: German Government Retrieved from http://www.bfr.bund.de/de/presseinformation/2012/29/studie_der_universitaet_caen_ist_kein_anlass_fuer_eine_neube_wertung_von_glyphosat _und_gentechnisch_veraendertem_mais_nk_603-131728.html AND http://www.ask-force.org/web/Seralini/BfR-bund-Stellungsnahme-Seralini- 20120928.pdf AND http://www.bfr.bund.de/cm/343/veroeffentlichung-von-seralini-et-al-zu-einer-fuetterungsstudie-an-ratten-mit- gentechnischveraendertem-mais-nk603-sowie-einer-glyphosathaltigen-formulierung.pdf AND English http://www.ask-force.org/web/Seralini/BfR- bund-Position-engl-Seralini-20121001.pdf.

Bohannon John. (20151119). Feature: How to hijack a journal. Science Magazine, pp. 4. http://retractionwatch.com/2015/11/19/can-journals-get- hijacked-apparently-yes/ AND http://www.ask-force.org/web/Peer-Review/Bohannon-How-to-hijack-a-journal-20151119.pdf

Bordoni, R., Mezzelani, A., Consolandi, C., Frosini, A., Rizzi, E., Castiglioni, B., Salati, C., Marmiroli, N., Marchelli, R., Bernardi, L. R., Battaglia, C., & De Bellis, G. (2004). Detection and quantitation of genetically modified maize (Bt-176 Transgenic maize) by applying ligation detection reaction and universal array technology. Journal of Agricultural and Food Chemistry, 52(5), pp. 1049-1054. ://000220039800005

Brake, J., & Vlachos, D. (1998). Evaluation of transgenic event 176 "Bt" corn in broiler chickens. Poultry Science, 77(5), pp. 648-653. ://000073345900003

Brenner, B. M. (1985). NEPHRON ADAPTATION TO RENAL INJURY OR ABLATION. American Journal of Physiology, 249(3), pp. F324-F337. ://WOS:A1985AQW2900053 AND http://www.ask-force.org/web/Seralini/Brenner-Nephron-Adptation-Renl-Injury-1985.pdf

Brusetti, L., Francia, P., Bertolini, C., Pagliuca, A., Borin, S., Sorlini, C., Abruzzese, A., Sacchi, G., Viti, C., Giovannetti, L., Giuntini, E., Bazzicalupo, M., & Daffonchio, D. (2004). Bacterial communities associated with the rhizosphere of transgenic Bt 176 maize (Zea mays) and its non transgenic counterpart. Plant and Soil, 266(1-2), pp. 11-21. ://000226385500003 AND http://www.ask-force.org/web/Bt/Brusetti-Bacterial- communities-2004.pdf

Butler Declan. (20120925). Rat study sparks GM furore, Cancer claims put herbicide-resistant transgenic maize in the spotlight. Nature, 489(News in Focus), pp. 484. http://www.nature.com/news/rat-study-sparks-gm-furore-1.11471 AND http://www.ask-force.org/web/Seralini/Butler-Rat-Study- Sparks-GM-Furore-2012.pdf

CERA (AGBIOS). (2012). GM Crop Database. Center for Environmental Risk Assessment (CERA),. from

Chapin, R. E., Boekelheide, K., Cortvrindt, R., van Duursen, M. B. M., Gant, T., Jegou, B., Marczylo, E., van Pelt, A. M. M., Post, J. N., Roelofs, M. J. E., Schlatt, S., Teerds, K. J., Toppari, J., & Piersma, A. H. (2013). Assuring safety without animal testing: The case for the human testis in vitro. Reproductive Toxicology, 39, pp. 63-68. ://WOS:000320678700009 AND http://www.ask-force.org/web/Seralini/Chapin-Assuring-Safety- Animals-2013.pdf

Chassy, B., & Parrott, W. (2009). Is This Study Believable? Examples from Animal Studies with GM Foods. . In Agricultural Biotechnology, , pp. 9. http://www.agribiotech.info/details AND http://www.ask-force.org/web/Peer-Review/Chassy-Parrott-Believable-2009.doc

Chassy, B. M. (2009). Global Regulation of Transgenic Crops Molecular Genetic Approaches to Maize Improvement (Ch. pp.107-124) http://dx.doi.org/10.1007/978-3-540-68922-5_9

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Chassy, B. M., & Miller, H. I. (20120222). The Science of Things That Aren't So Forbes, February 2012, pp. http://www.forbes.com/sites/henrymiller/2012/02/22/the-science-of-things-that-arent-so/ AND http://www.ask-force.org/web/Seralini/Chassy- Miller-Seralini-Rebuttal-Forbes-20120222.pdf

Chiueh, L. C., Lin, C. Y., Chen, Y. C., Wu, M. H., & Shih, D. Y. C. (2004). Establishment of quantitative method for five events of genetically modified maize (Bt11, Event176, GA21, MON810 and T25) by real-time QPCR. Journal of Food and Drug Analysis, 12(4), pp. 316-323. ://000225988300005

Clair, E., Linn, L., Travert, C., Amiel, C., Seralini, G.-E., & Panoff, J.-M. (2012). Effects of Roundup (R) and Glyphosate on Three Food Microorganisms: Geotrichum candidum, Lactococcus lactis subsp. cremoris and Lactobacillus delbrueckii subsp. bulgaricus. Current Microbiology, 64(5), pp. 486-491. ://WOS:000301841000012 AND http://www.ask-force.org/web/Seralini/Clair-Effects-Roundup-Glyphosate- Microorganisms-2012.pdf

Clair, É., Mesnage, R., Travert, C., & Séralini, G.-É. (2012). A glyphosate-based herbicide induces necrosis and apoptosis in mature rat testicular cells in vitro, and testosterone decrease at lower levels. Toxicology in Vitro, 26(2), pp. 269-279. http://www.sciencedirect.com/science/article/pii/S0887233311003341

CNRS (Producer). (20120927). Pétition Pour un débat raisonné sur les OGM. Retrieved from http://www.cnrs.fr/fr/une/actus/2012/20120927- debat-ogm.html AND http://www.ask-force.org/web/Seralini/CNRS-Petition-Seralini-20120927.pdf

Cockburn, A. (20121107). Letter to the Editor. Food and Chemical Toxicology(0), pp. http://www.sciencedirect.com/science/article/pii/S0278691512007855 AND http://www.ask-force.org/web/Seralini/Cockburn-Letter-to-Editor- 20121107.pdf

Collotta, M., Bertazzi, P. A., & Bollati, V. (2013). Epigenetics and Pesticides. Toxicology(0), pp. http://www.sciencedirect.com/science/article/pii/S0300483X1300022X AND http://www.ask-force.org/web/Seralini/Collota-Epigenetics- Pesticides-Sprague-2013.pdf

COMETS-CNRS. (20121002). Rappels du COMETS sur les aspects éthiques de la communication des chercheurs avec les média pp. Paris ISBN/ISSN Le Comité d'éthique du CNRS (COMETS), http://www.cnrs.fr/fr/organisme/ethique/comets/index.htm Retrieved from http://www.cnrs.fr/fr/organisme/ethique/comets/docs/121003-rappel-deontologie-ong.pdf AND http://www.ask-force.org/web/Seralini/COMETS- Rappel-Deontologie-Communication-20121002.pdf

Conti, B., Maltoni, C., Perino, G., & Ciliberti, A. (1988, online 2006). LONG-TERM CARCINOGENICITY BIOASSAYS ON STYRENE ADMINISTERED BY INHALATION, INGESTION AND INJECTION AND STYRENE OXIDE ADMINISTERED BY INGESTION IN SPRAGUE-DAWLEY RATS, AND PARA- METHYLSTYRENE ADMINISTERED BY INGESTION IN SPRAGUE-DAWLEY RATS AND SWISS MICE. Annals of the New York Academy of Sciences, 534, pp. 203-234. ://WOS:A1988Q080100021 AND online 2006 published http://onlinelibrary.wiley.com/doi/10.1111/j.1749- 6632.1988.tb30112.x/pdf AND http://www.ask-force.org/web/Seralini/Conti-Long-Term-Carcinogenicity-Bioassays-Styrene-2006.pdf

Corbisier, P., Broothaerts, W., Gioria, S., Schimmel, H., Burns, M., Baoutina, A., Emslie, K. R., Furui, S., Kurosawa, Y., Holden, M. J., Kim, H. H., Lee, Y., Kawaharasaki, M., Sin, D., & Wang, J. (2007). Toward metrological traceability for DNA fragment ratios in GM quantification. 1. Effect of DNA extraction methods on the quantitative determination of Bt176 corn by real-time PCR. Journal of Agricultural and Food Chemistry, 55(9), pp. 3249-3257. ://WOS:000245946300003

Council-Biotechnology. (20120920). Scientists in U.S. Reject Heavily Publicized Rat Study, Other Studies Show No Impact of Feeding Biotech Grain pp. Washington DC, USA ISBN/ISSN Retrieved from www.whybiotech.com AND http://www.ask- force.org/web/Seralini/Council-Biotech-Seralini-20120920.pdf

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CRIIGEN, & Seralini Gilles-Eric. (2012). FAQ – From CRIIGEN research Team, Protocole - Results - Discussion, SECTION ONE : PROTOCOL, SECTION TWO RESULTS, SECTION THREE DISCUSSION. from http://www.criigen.org/SiteEn/index.php?option=com_content&task=view&id=368&Itemid=1 AND http://www.ask-force.org/web/Seralini/CRIIGEN-Seralini-FAQs-Protocol-2012.pdf see more: http://www.criigen.org/SiteEn/

CTNBio, José Fernando Garcia, Fernando Salvador Moreno, & Nance Beyer Nardi. (20121024). Evaluation Avaliacao de Publicacao de Trabalho Experimental em Carater de Urgencia, Considered Opinion on Seralini Publication pp. 10 Brasilia Brazil: CTNBio ISBN/ISSN CTNBio Ministerio de Ciencia Technologia e Inovacao, Brazilian National Technical Commission on Biosafety Retrieved from http://www.ask-force.org/web/Seralini/CTNBio-Seralini-Avaliacao-port-20121021.pdf AND http://www.ask-force.org/web/Seralini/CTNbio-Seralini- Evaluation-en-.20121024.pdf

de Liz Oliveira Cavalli, V. L., Cattani, D., Heinz Rieg, C. E., Pierozan, P., Zanatta, L., Benedetti Parisotto, E., Wilhelm Filho, D., Mena Barreto Silva, F. R., Pessoa-Pureur, R., & Zamoner, A. (2013). Roundup disrupts male reproductive functions by triggering calcium-mediated cell death in rat testis and Sertoli cells. Free Radical Biology and Medicine, 65(0), pp. 335-346. http://www.sciencedirect.com/science/article/pii/S0891584913003262

de Melo-Martin, I., & Meghani, Z. (2008). Beyond risk - A more realistic risk - benefit analysis of agricultural biotechnologies. Embo Reports, 9(4), pp. 302-306. ://WOS:000254561300002 AND http://www.ask-force.org/web/Regulation/DeMelo-Martin-Beyond-Risk-EMBO-2008.pdf

de Souza, L., & Macedo Oda, L. (2013). Letter to the editor. Food and Chemical Toxicology, 53(0), pp. 440. http://www.sciencedirect.com/science/article/pii/S0278691512008022

de Vendomois, J. S., Roullier, F., Cellier, D., & Seralini, G. E. (2009). COMPARISON OF THE EFFECTS OF THREE GM CORN ON MAMMALIAN HEALTH International Journal of Biological Sciences, 5, pp. 706-721. http://www.biolsci.org/v05p0706 AND http://www.ask- force.org/web/Food/DeVentomois-Comparison-Effects-Three-2009.pdf

Dietrich, D., von Aulock, S., Marquardt, H. W. J., Blaauboer, B. J., Dekant, W., Kehrer, J., Hengstler, J. G., Collier, A. C., Gori, G. B., Pelkonen, O., Lang, F., Nijkamp, F. P., Stemmer, K., Li, A., Savolainen, K., Hayes, A. W., Gooderham, N., & Harvey, A. (2013). Open letter to the European commission: scientifically unfounded precaution drives European commission's recommendations on EDC regulation, while defying common sense, well-established science, and risk assessment principles. Archives of Toxicology, 87(9), pp. 1739-1741. ://WOS:000323734900010 AND http://www.ask-force.org/web/Evolution/Dietrich-Open-Letter-EC-Scientifically-unfounded-2013.pdf

Dietrich, D. R., von Aulock, S., Marquardt, H., Blaauboer, B., Dekant, W., Kehrer, J., Hengstler, J., Collier, A., Gori, G. B., Pelkonen, O., Lang, F., Nijkamp, F. P., Stemmer, K., Li, A., Savolainen, K., Hayes, A. W., Gooderham, N., & Harvey, A. (2013). Scientifically unfounded precaution drives European Commission's recommendations on EDC regulation, while defying common sense, well-established science and risk assessment principles. Toxicology in Vitro, 27(7), pp. 2110-2114. ://WOS:000326361000011 AND http://www.ask-force.org/web/Regulation/Dietrich-et-al- Letter-Scientifically-Unfounded-Reg-Toxicol-Pharmacol-2013.pdf

Dinse, G. E., Peddada, S. D., Harris, S. F., & Elmore, S. A. (2010). Comparison of NTP Historical Control Tumor Incidence Rates in Female Harlan Sprague Dawley and Fischer 344/N Rats. Toxicologic Pathology, 38(5), pp. 765-775. http://tpx.sagepub.com/content/38/5/765.abstract AND http://www.ask-force.org/web/Seralini/Dinse-Comparison-NTP-Historical-Control-Tumor-2010.pdf

Domingo, J. L. (2000). Health risks of GM foods: Many opinions but few data. Science, 288(5472), pp. 1748-1749. ://000087503800023 AND http://www.ask-force.org/web/Food/Domingo-Health-Risks-Science-2000.PDF

Domingo, J. L. (2007). Toxicity studies of genetically modified plants: A review of the published literature. Critical Reviews in Food Science and Nutrition, 47(8), pp. 721-733. ://000251214200003 AND http://www.ask-force.org/web/Bt/Domingo-Toxicity-Studies-2007.pdf

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Edwards, I. B., & Beetles, C. (2012). Information about Australian Tour of G.E. Seralini in February 2012 Newspaper, Report and Fake Diploma. Stock & Land, . Retrieved from http://www.ask-force.org/web/Seralini/Edwards-Beetles-Seralini-Information-Sheet-Australia-2012.pdf see comment in French by Marcel Kunzt: http://www.agriculture-environnement.fr/a-la-une,6/seralini%E2%80%89-le-prix-de-l-ego,771.html

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EFSA-Opinion. (2006). Opinion of the Scientific Panel on genetically modified organisms [GMO] related to genetically modified crops (Bt176 maize, MON810 maize, T25 maize, Topas 19/2 oilseed rape and Ms1xRf1 oilseed rape) subject to safeguard clauses invoked according to Article 16 of Directive 90/220/EEC Opinion adopted on 29 March 2006, Publication date: 11/04/2006 Last updated: 27/07/2006 The EFSA Journal, 338, pp. 1-15. http://www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1178620769622.htm AND http://www.ask-force.org/web/Regulation/EFSA- Opinion-Maize-Bt176-MON810-t25-2006.pdf

EFSA-Statement. (2007). Statement of the Scientific Panel on Genetically Modified Organisms on the analysis of data from a 90-day rat feeding study with MON 863 maize pp. 5 ISBN/ISSN European Food Safety Authority Retrieved from http://www.efsa.europa.eu/EFSA/Statement/GMO_statement_MON863,0.pdf

EFSA-Statement. (20121004). Review of the Séralini et al. (2012) publication on a 2-year rodent feeding study with glyphosate formulations and GM maize NK603 as published online on 19 September 2012 in Food and Chemical Toxicology EFSA Journal, 2012;10(10):2910 pp. 1-9. Press release: http://www.efsa.europa.eu/en/press/news/121004.htm AND Assessment: http://www.efsa.europa.eu/en/efsajournal/doc/2910.pdf AND EFSA- Letter to Seralini: http://www.efsa.europa.eu/en/edinterviews/docs/corporateSeralini121004.pdf AND http://www.ask- force.org/web/Regulation/EFSA.Statement-Seralini-Journal-201210-10-2912-20121004.pdf

EFSA. (2007a). Press release: EFSA reaffirms its risk assessment of genetically modified maize MON 863 maize pp. 5 ISBN/ISSN European Food Safety Authority Retrieved from http://www.ask-force.org/web/Food/EFSA-Reaffirmation-mon863-20070628.pdf

EFSA. (2007b). Safety and nutritional assessment of GM plants and derived food and feed: The role of animal feeding trials. Safety and nutritional assessment of GM plants and derived food and feed: The role of animal feeding trials, 46(Supplement 1), pp. S2-S70. http://www.sciencedirect.com/science/article/B6T6P-4RTW3XD-1/1/3bf8f16f11a571d65ea649c1260bb3da

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EFSA. (20151006). Review of results published by Mesnage et al. (2015) in PLoS ONE and the laboratory findings communicated by Dr Samsel to Farm Wars. European Food Safety Authority. EJ EFSA Journal Scientific Report, pp. doi:10.2903/j.efsa.2015.4258 AND http://www.efsa.europa.eu/sites/default/files/scientific_output/files/main_documents/4258.pdf AND http://www.ask- force.org/web/Seralini/EFSA-Review-Mesnage-PLoS-ONE-and-Laboratory-findings-Samsel-Farm-Wars-20151002.pdf

EFSA Independence. (20121028a). On the Indepencence of EFSA. 20121028, from http://www.youtube.com/watch?v=N0FEVglpQcw&feature=player_embedded AND http://www.efsa.europa.eu/en/topics/topic/independence.htm AND http://www.ask-force.org/web/Seralini/EFSA-Independence-Statements- 20121028.pdf

EFSA Independence. (20121028b). On the Indepencence of EFSA. Collection of 32 website links on the Independence of EFSA. 20121028, from http://www.youtube.com/watch?v=N0FEVglpQcw&feature=player_embedded AND http://www.efsa.europa.eu/en/topics/topic/independence.htm AND http://www.ask-force.org/web/Seralini/EFSA-Independence-Statements- 20121028.pdf

EFSA News. (20121218). Commission and EFSA agree need for two-year GMO feeding studies. from www.gmwatch.org/index.php?option=com_content&view=article&id=14509 AND http://www.ask- force.org/web/EFSA/EFSA-agree-two-year-GMO-feeding-studies-20121218.pdf

EFSA Opinion. (20110524). Guidance for risk assessment of food and feed from genetically modified plants. EFSA Journal, 9(5), pp. 37. http://www.efsa.europa.eu/en/efsajournal/pub/2150.htm AND http://www.ask-force.org/web/EFSA/EFSA-Opinion-Guidance-Risk-Assessment- 20110524.pdf

EFSA Statement. (20121004). Review of the Séralini et al. (2012) publication on a 2-year rodent feeding study with glyphosate formulations and GM maize NK603 as published online on 19 September 2012 in Food and Chemical Toxicology EFSA Journal, 2012;10(10):2910 pp. 1-9. Press release: http://www.efsa.europa.eu/en/press/news/121004.htm AND Assessment: http://www.efsa.europa.eu/en/efsajournal/doc/2910.pdf AND EFSA- Letter to Seralini: http://www.efsa.europa.eu/en/edinterviews/docs/corporateSeralini121004.pdf AND http://www.ask- force.org/web/Regulation/EFSA.Statement-Seralini-Journal-201210-10-2912-20121004.pdf

EFSA Statement. (20121123). Final review of the Séralini et al. (2012a) publication on a 2-year rodent feeding study with glyphosate formulations and GM maize NK603 as published online on 19 September 2012 in Food and Chemical Toxicology. EFSA Journal, 10(11, 2986), pp. 1-10. http://www.efsa.europa.eu/en/efsajournal/doc/2986.pdf AND http://www.ask-force.org/web/HerbizideTol/Seralini-EFSA_2986-annex-1-2012.pdf AND http://www.ask-force.org/web/HerbizideTol/EFSA-final-Review-Seralini--2986-20121123.pdf AND http://www.ask- force.org/web/HerbizideTol/EFSA-Seralini-Final-Press-Release-20121128.pdf

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El-Sanhoty, R., Shahwan, T., & Ramadan, M. F. (2006). Application of artificial neural networks to develop a classification model between genetically modified maize (Bt-176) and conventional maize by applying lipid analysis data. Journal of Food Composition and Analysis, 19(6-7), pp. 628-636. ://000238776500022

Elsevier, Hatton Jackie, John Schrefer, Peggy Bridges, Angelika Lex, Claudio Rothmuller, Celeste Giampetro, James Phimister, and, W. T., & Ryoji Fukada. (2005). A short history of Elsevier, in celebration of the 125th of Elsevier and the 425th anniversary of the House of Elzevir pp. 10 Netherlands, Amsterdam ISBN/ISSN Elsevier Retrieved from http://www.elsevier.com/about/history AND http://www.ask- force.org/web/Seralini/Elsevier-Short-History-2005.pdf

ENSSER Statement Seralini. (20121005). Questionable Biosafety of GMOs, Double Standards and, once again, a "Shooting-the-Messenger" style Debate pp. 8 ENSSER Statement on Séralini et al. (2012) publication and reactions evoked Hyderabad, India at COP-6 Meeting ISBN/ISSN ENSSER, European Network of Scientists for Social and Environmental Responsibility Retrieved from http://xa.yimg.com/kq/groups/18208928/312025102/name/ENSSER_Seralini.pdf AND http://www.ask-force.org/web/Seralini/ENSSER-Statement- Seralini-20121005.pdf

Entine Jon. (20130610). Being Gilles-Eric Seralini: Inside the mind of the anti-GM movement. Genetic Literacy Project,, pp. 6. https://www.geneticliteracyproject.org/2013/06/10/being-gilles-eric-seralini-inside-the-mind-of-the-anti-gm-movement/ AND http://www.ask- force.org/web/Seralini/Entine-Being-Gilles-Eric-Seralini-Inside-mind-20130610.pdf

EuroBioNews. (20121012). French Study Suggests GMOs Cause Health Problems. European Biotechnology News, 11(Issue 10), pp. 1 (28). http://www.ask-force.org/web/Seralini/EuroBioNews-French-Study-Suggests-GMOs-201210.pdf

EuropaBio (Producer). (20120919). Key messages – Seralini’s study, Key Messages. Retrieved from http://www.ask- force.org/web/Seralini/EUROPABIO-Key-messages-Seralinistudy-UK-20120919-2.pdf

EUSJA Board. (20121005). EUSJA Statement on embargoes and manipulation pp. Bad Gastein, Germany ISBN/ISSN EUSJA, Europen Union of Science Journalist Association Retrieved from http://networkedblogs.com/D5nZ0 AND http://www.ask-force.org/web/Seralini/EUSJA-Statement- Embargoes-20121005.pdf

Evers Elsa, & Greenpeace. (2012). Gilles-Eric Séralini Biography Australia Tour, Greenpeace. pp. http://www.greenpeace.org/australia/Global/australia/True_food/Seralini_Bio.pdf AND http://www.ask-force.org/web/Seralini/Evers-Greenpeace- Seralini-Bio-Australia-Tour-2012.pdf

Fang, F. C., & Casadevall, A. (2012). Reply to "Likelihood of False-Positive Results in High-Impact Journals Publishing Groundbreaking Research". Infection and Immunity, 80(3), pp. 1301. http://iai.asm.org/content/80/3/1301.short AND http://www.ask-force.org/web/Peer-Review/Fang-Reply- Likelyhood-High-Impact-2012.pdf

Fang, F. C., Steen, R. G., & Casadevall, A. (2012). Misconduct accounts for the majority of retracted scientific publications. Proceedings of the National Academy of Sciences, 109(42), pp. 17028-17033. http://www.pnas.org/content/109/42/17028.abstract AND http://www.ask- force.org/web/Peer-Review/Fang-Misconduct-Accounts-Majority-retracted-2012.pdf

Felke, M., Langenbruch, G.-A., Feiertag, S., & Kassa, A. (2010). Effect of Bt-176 maize pollen on first instar larvae of the Peacock butterfly (Inachis io) (Lepidoptera; Nymphalidae). Environmental. Biosafety Research, -- Received: 20 November 2008, Accepted: 5 December 2009, online publication 28. October 2010 (!!)(--), pp. --. http://dx.doi.org/10.1051/ebr/2010006 AND http://www.ask-force.org/web/Bt1/Felke-Effect-Bt-176-Peacock- 2010.pdf

Feriotto, G., Gardenghi, S., Bianchi, N., & Gambari, R. (2003). Quantitation of Bt-176 maize genomic sequences by surface plasmon resonance- based biospecific interaction analysis of multiplex polymerase chain reaction (PCR). Journal of Agricultural and Food Chemistry, 51(16), pp. 4640- 4646. ://000184356700021 49

Ferrini, A. M., Mannoni, V., Pontieri, E., & Pourshaban, M. (2007). Longer resistance of some DNA traits from BT176 maize to gastric juice from gastrointestinal affected patients. International Journal of Immunopathology and Pharmacology, 20(1), pp. 111-118. ://WOS:000245120600013

Flamm, W. G., Blackburn, G. L., Comer, C. P., Mayhew, D. A., & Stargel, W. W. (2003). Long-term food consumption and body weight changes in neotame safety studies are consistent with the allometric relationship observed for other sweeteners and during dietary restrictions. Regulatory Toxicology and Pharmacology, 38(2), pp. 144-156. http://www.sciencedirect.com/science/article/pii/S0273230003000758 AND http://www.ask- force.org/web/Seralini/Flamm-Long-Term-Food-Consumption-Sweeteners-2003.pdf

Fogo, A., & Ichikawa, I. (1989). EVIDENCE FOR THE CENTRAL ROLE OF GLOMERULAR GROWTH PROMOTERS IN THE DEVELOPMENT OF SCLEROSIS. Seminars in Nephrology, 9(4), pp. 329-342. ://WOS:A1989CC86100004

Folta, K. (2014). Letter to the Editor. Food and Chemical Toxicology(0), pp. http://www.sciencedirect.com/science/article/pii/S0278691514000052 AND http://www.ask-force.org/web/Seralini/Folta-Kevin-Letter-Editor-2014.pdf

Frazier, K. S., Seely, J. C., Hard, G. C., Betton, G., Burnett, R., Nakatsuji, S., Nishikawa, A., Durchfeld-Meyer, B., & Bube, A. (2012). Proliferative and Nonproliferative Lesions of the Rat and Mouse Urinary System. Toxicologic Pathology, 40(4 suppl), pp. 14S-86S. http://tpx.sagepub.com/content/40/4_suppl/14S.abstract AND http://www.ask-force.org/web/Seralini/Frazier-Proliferative-Nonproliferative- Lesions-Urinary-2012.pdf

Fresco, L. O. (2013). The GMO Stalemate in Europe. Science, 339(6122), pp. 883. http://www.sciencemag.org/content/339/6122/883.short AND http://www.ask-force.org/web/Seralini/Fresco-GMO-Stalemate-Europe-Science-20130222.pdf

Friston, K. (2012). Ten ironic rules for non-statistical reviewers. NeuroImage, 61(4), pp. 1300-1310. http://www.sciencedirect.com/science/article/pii/S1053811912003990

FSANZ Food Standard A-NZ. (20121011). Response (preliminary) to Séralini paper on the long term toxicity of a Roundup herbicide and a Roundup- tolerant genetically modified maize Retrieved 1, 33, from http://www.foodstandards.gov.au/consumerinformation/gmfoods/gmfactsheets/responsetosralinipap5676.cfm AND http://www.ask- force.org/web/Seralini/FSANZ-Seralini-Preliminary-Response-201210.pdf

Garcia-Canas, V., Cifuentes, A., & Gonzalez, R. (2004). Quantitation of transgenic Bt event-176 maize using double quantitative competitive polymerase chain reaction and capillary gel electrophorsesis laser-induced fluorescence. Analytical Chemistry, 76(8), pp. 2306-2313. ://000221096800020

Gardenghi, S., Finotti, A., Gambari, R., & Feriotto, G. (2004). Real-time detection of genetically modified maize Bt-176 genomic sequences by surface plasmon resonance-based biospecific interaction analysis. Minerva Biotecnologica, 16(3), pp. 203-210. ://000225250200005

Gasnier, C., Dumont, C., Benachour, N., Clair, E., Chagnon, M. C., & Seralini, G. E. (2009). Glyphosate-based herbicides are toxic and endocrine disruptors in human cell lines. Toxicology, 262(3), pp. 184-191. ://WOS:000269287700002 AND http://www.ask- force.org/web/Seralini/Gasnier-Glyphosate-based-Herbicides-2009.pdf

Gasnier, C., Laurant, C., Decroix-Laporte, C., Mesnage, R., Clair, E., Travert, C., & Seralini, G.-E. (2011). Defined plant extracts can protect human cells against combined xenobiotic effects. Journal of occupational medicine and toxicology (London, England), 6(1), pp. 3-3. ://MEDLINE:21251308 AND http://www.ask-force.org/web/Seralini/Gasnier-Defined-Plant-Extracts-2011.pdf

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Gloeckner Gottfried, & Seralini Gilles-Eric. (20160122). Pathology reports on the first cows fed with Bt176 maize (1997–2002) and short comment. Scholarly Journal of Agricultural Science (the domain no more available since January 27), 6(1), pp. 1-8. This domain name expired on Jan 27 2015 06:39PM AND http://www.ask-force.org/web/Seralini/Gloeckner-Seralini-Pathology-Reüprts-Bt176-Maize-Cows-201601.pdf AND short comment http://www.ask-force.org/web/Seralini/Seralini-Experience-first-GM-crop-Europe-short-20160122.pdf

GM-Free-Cymru. (20121004). EFSA slammed for "institutionalised pseudo-science" after rat study review from http://www.gmfreecymru.org/documents/efsa_slammed.html AND http://www.ask-force.org/web/Seralini/GM-Free-Cymru-EFSA-Slammed- 20121004.pdf

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Wager Rob, Giddings Val, signed by, Ammann Klaus, Bradford Kent, Burachik Moises, Cetiner Selim, Chassy Bruce M., De Souza Lucia, De Stefano-Beltran Louis, Fellous Marc, Fedoroff Nina, Herring Ron, Klaus, J., Leaver Chris, Lerayer Alda, McHughe Alan, I., M. H., Newell- McGloughlin Martina, Potrykus Ingo, CS, P., Rao Kameswara, Sabah, A., Sivramiah, S., Strauss Steven, & Trewavas Antony. (20120924 and 2013). Letter to the Editor in Chief of Food, Chemical Toxicology (Elsevier), to request a serious reconsideration of the recent paper by Seralini et al. alleging tumorigenesis in rats resulting from consumption of corn derived from crops improved through biotechnology. pp. 1-4. http://www.ask- force.org/web/Seralini/Wager-Letter-Editor-Hayes-Seralini-20120925.pdf

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