Biodegradation of synthetic polymers in soils: Tracking carbon into CO2 and microbial biomass


Plastic materials are widely used in agricultural applications to achieve food security for the growing world population. The use of biodegradable instead of nonbiodegradable polymers in single-use agricultural applications, including plastic mulching, promises to reduce plastic accumulation in the environment. We present a novel approach that allows tracking of carbon from biodegradable polymers into CO2 and microbial biomass. The approach is based on 13C-labeled polymers and on isotope-specific analytical methods, including nanoscale secondary ion mass spectrometry (NanoSIMS). Our results unequivocally demonstrate the biodegradability of poly(butylene adipate-co-terephthalate) (PBAT), an important polyester used in agriculture, in soil. Carbon from each monomer unit of PBAT was used by soil microorganisms, including filamentous fungi, to gain energy and to form biomass. This work advances both our conceptual understanding of polymer biodegradation and the methodological capabilities to assess this process in natural and engineered environments.


Modern agriculture heavily relies on the use of plastic materials in various applications, a practice coined plasticulture. Mulching with plastic films is a major application with a global market volume of approximately 2 × 106 tons per year (1). Mulch films are placed onto agricultural soils to improve conditions for plant growth while lowering consumption of water, herbicides, and fertilizer and also minimizing soil erosion (1, 2). Because of these benefits, mulching with plastic films helps to secure food for the growing world population. However, mulch films are commonly composed of nonbiodegradable polyethylene and, therefore, accumulate in agricultural soils and surrounding receiving environments if incompletely retrieved after use. These accumulations have negative ecologic and economic impacts, including decreased soil productivity (35). A promising strategy to overcome these risks is to use mulch films composed of polymers that biodegrade in soils (1, 68).

Biodegradation of polymers requires microorganisms to metabolize all organic components of the polymer. Biodegradation in soil involves several key steps (Fig. 1): (i) colonization of the polymer surface by microorganisms, (ii) secretion of extracellular microbial enzymes that depolymerize the polymer into low–molecular weight compounds, and (iii) microbial uptake and utilization of these compounds, incorporating polymer carbon into biomass or releasing it as CO2 (9).

Fig. 1 Key steps in the biodegradation of polymers in soils.

Microorganisms colonize the polymer surface and secrete extracellular enzymes that depolymerize the polymer. The formed low–molecular weight hydrolysis products are taken up by the microorganisms and used both for energy production, resulting in the formation of CO2, and for the synthesis of cellular structures and macromolecules, resulting in incorporation of polymer-derived carbon into the microbial biomass. The boxes on the right depict the analytical methods we used to study these steps. NMR, nuclear magnetic resonance.

Fig. 1 Key steps in the biodegradation of polymers in soils.Microorganisms colonize the polymer surface and secrete extracellular enzymes that depolymerize the polymer. The formed low–molecular weight hydrolysis products are taken up by the microorganisms and used both for energy production, resulting in the formation of CO2, and for the synthesis of cellular structures and macromolecules, resulting in incorporation of polymer-derived carbon into the microbial biomass. The boxes on the right depict the analytical methods we used to study these steps. NMR, nuclear magnetic resonance.Here, we examined each of the above steps for poly(butylene adipate-co-terephthalate) (PBAT), an aliphatic-aromatic statistical copolyester of large importance in the market of biodegradable mulch films (7). While previous studies provided indirect indications for PBAT biodegradation in soils based on determining PBAT mass loss and changes in its physicochemical properties (1012), we here use a novel workflow using stable carbon isotope-labeled PBAT to directly and unequivocally demonstrate its biodegradation in soil (table S1). This workflow included incubation of 13C-labeled polymer films in soil with continuous quantification of polymer-derived 13CO2 by isotope-specific cavity ring-down spectroscopy (CRDS) (13). The use of 13C-labeled polymers allowed us to distinguish polymer-derived CO2 from CO2formed by soil organic matter mineralization. After incubation, we imaged the polymer film surfaces using scanning electron microscopy (SEM) and demonstrated the incorporation of polymer-derived 13C into the biomass of film-colonizing microorganisms using element-specific, isotope-selective nanoscale secondary ion mass spectrometry (NanoSIMS) (14). We studied three PBAT variants that had similar physicochemical properties and comparable total 13C contents, but varied in the monomer that contained the 13C-label [that is, butanediol (P*BAT), adipate (PB*AT), or terephthalate (PBA*T)] (Fig. 2A and table S2). The use of these variants allowed us to follow biodegradation of all PBAT building blocks. The presented workflow is a novel approach to study the fundamental steps in polymer biodegradation in complex systems (1517).


This work presents an experimental approach to study polymer biodegradation in soils and to assess the key steps involved in this process: microbial polymer colonization, enzymatic depolymerization on the polymer surface, and microbial uptake and utilization of the released low–molecular weight compounds. Central to the approach is the use of polymer variants that are 13C-labeled in all monomer units of the polymer, thereby allowing us to assess whether all organic components of the polymer material are used by soil microorganisms. The label further allows tracing of polymer-derived carbon into both CO2 and microbial biomass. Using this approach, we demonstrate here the biodegradability of PBAT in soil. Biodegradability renders PBAT a more environmentally friendly alternative to persistent polymer materials for use in plasticulture, including single-use applications such as plastic mulching. Our results further imply that incorporation of polymer-derived carbon into microbial biomass needs to be taken into consideration in regulatory guidelines for determining biodegradability of polymers. Currently, these guidelines are solely based on extents of CO2 formation. Furthermore, the finding of subcellular structures within PBAT-colonizing fungi highly enriched in polymer-derived carbon might represent compartments in which carbon is stored (for example, in the form of neutral lipids) when fungi are limited by the availability of nutrients other than carbon (22). These limitations are plausible for microorganisms growing on PBAT and other polymers that do not contain nitrogen and phosphorous. If these limitations occur, increasing the availability of soil nutrients to microorganisms colonizing the polymer surface is expected to enhance polymer biodegradation.

This work demonstrates PBAT biodegradation in a selected agricultural soil over 6 weeks of incubation. Future studies extending on this work will need to assess variations in the rates and extents of PBAT mineralization among different agricultural soils, also over longer-time incubations. Furthermore, we propose studies that are directed toward identifying soil microorganisms that are actively involved in PBAT biodegradation. While the NanoSIMS-based approach presented here allows us to unambiguously demonstrate incorporation of polyester carbon into soil microbial biomass, it is not a high-throughput technique. Alternative approaches, including the extraction of targeted biomolecules from soils containing 13C-labeled polymers followed by quantifying the 13C contents in the extracted molecules, will allow us to analyze larger sample sets and thereby to systematically determine potential variations among soil microorganisms in the extent to which they incorporate polymer-derived carbon into their biomass.


Experimental design

The objective of this study was to develop an experimental approach to demonstrate biodegradation of PBAT in an agricultural soil. As biodegradation includes mineralization of PBAT carbon to CO2, as well as the incorporation of PBAT-derived carbon into the biomass of soil microorganisms, we addressed both of these processes in controlled laboratory experiments. We followed PBAT mineralization during soil incubation using an isotope-specific CRDS for the quantification of formed CO2. For each of the three PBAT variants, we simultaneously incubated seven films in one incubation bottle filled with soil to allow precise quantification of PBAT mineralization to CO2. The soil incubations were terminated after 6 weeks (that is, when approximately 10% of the PBAT carbon had been mineralized) to ensure that PBAT films were still intact for the subsequent imaging analyses. We revealed incorporation of PBAT-derived carbon into biomass using NanoSIMS, which enabled identification of subcellular features and determination of the carbon isotope composition of the PBAT film surface and the colonizing microorganisms at submicrometer spatial resolution. The low throughput of this high-end topochemical analysis technique constrained the number of collected images for soil-incubated films to two images for each of the three PBAT variants including replicate films. We note that we did not exclude any data or outliers from our analysis.

Polyesters, monomers, soil, and enzymes

Polyesters were provided by BASF SE and synthesized as previously described (23, 24). The physicochemical properties of the polyesters are listed in table S2. To obtain similar 13C contents for the three PBAT variants (that is, PB*AT, P*BAT, and PBA*T), synthesis of all variants was performed with defined ratios of labeled to unlabeled monomers. The three PBAT variants were free of chemical additives.

The 13C-labeled monomers 1,6-13C2-adipate and 13C4-butanediol used for PBAT synthesis and for soil incubation studies were purchased from Sigma, with more than 99% of the indicated positions in the monomer containing 13C. We obtained 1-13C1-terephthalate from dimethyl 1-13C-terephthalate purchased from Sigma. To obtain the free diacid, we dissolved dimethyl 1-13C-terephthalate in 2:1 water/tetrahydrofuran (5 mg in 2.4 ml), added 25 μl of a sodium hydroxide solution [37% (w/w)], and stirred the solution at room temperature for 2 hours. The solvent was then carefully removed under reduced pressure to obtain the hydrolysis product 1-13C1-terephthalate (confirmed by 1H NMR).

For PBAT and monomer incubations in soils under controlled laboratory conditions, we used agricultural soils from the agricultural center Limburgerhof (Rhineland-Palatinate, Germany). Physicochemical properties of the soils are provided in table S1. The soils were air-dried to a humidity of 12% of the maximum water-holding capacity of the soil, passed through a 2-mm sieve, and stored in the dark at 4°C for 12 months before use in the incubation experiments.

R. oryzae lipase was purchased as a powder from Sigma (catalog no. 80612). FsC was obtained as a solution from ChiralVision B.V. (Novozym 51032). Stock solutions of both enzymes in water were stored at −20°C.

Preparation of PBAT films and soils for incubation experiments

We prepared two sets of solvent-cast PBAT films that differed in the way that the PBAT films were attached to the silicon wafer substrates. For the first set, we solvent-cast PBAT films by adding three times 15 μl of a PBAT solution in chloroform [concentration, 5% (w/w)] onto a square-cut antimony-doped silicon wafer platelet (7.1 mm × 7.1 mm × 0.75 mm, Active Business Company). In between the additions of the polymer solutions, we allow the chloroform to evaporate. This procedure resulted in a PBAT mass of approximately 3 mg per wafer. Before incubation in soil, the solvent-cast polyester films were stored in the dark at room temperature for 1 week to ensure complete evaporation of the solvent (chloroform). PBAT variants from this first set were used for PBAT mineralization experiments (Fig. 2B), SEM imaging (Fig. 2C), and NanoSIMS imaging (Figs. 3 and 4 and fig. S8).

For the second set of PBAT films, we pretreated the silicon wafer platelets with Vectabond (Vector Laboratories, catalog no. SP-1800) before solvent casting of the polyester films. This second set of PBAT films was included to test whether the adhesion of the PBAT to the Si surface can be improved by this modified protocol. For the pretreatment, we exposed the wafers to a 1:50 diluted solution of Vectabond in acetone, subsequently dipped them into MilliQ water (Barnstead Nanopure Diamond), and dried them in a stream of N2. PBAT variants from this set were used only to determine PBAT mineralization (fig. S1), but not for SEM and NanoSIMS imaging.

We prepared the soil for PBAT incubations by adding MilliQ water to the soil to adjust its water content to 47% of its maximum water-holding capacity. We subsequently transferred 60 g of the soil into each of the incubation vessels (100-ml glass Schott bottles). We prepared a total of nine incubation bottles in three sets of three bottles (see below). The soils were then preincubated at 25°C in the dark for 1 week.

After soil preincubation, we transferred the wafers carrying the solvent-cast polyester films into the soils in the incubation bottles. We added seven wafers to each incubation bottle. The wafers were spaced apart by at least 1 cm. All wafers were positioned upright in the soil. The three bottles of the first set each contained films of one of the three differently labeled PBAT variants obtained by direct solvent casting. The three bottles of the second set were identical to the first set except for the wafers, which were pretreated with Vectabond before solvent casting. The three bottles in the third set served as controls and contained soil but no PBAT films. All bottles were incubated for 6 weeks at 25°C in the dark. We note that our study therefore does not address potential effects of ultraviolet irradiation–induced changes in the structure of PBAT on its biodegradability. Over the course of the incubation, we gravimetrically determined the water content of the soils at defined time intervals. To sustain a constant soil water content, amounts of water that were lost from the soil through evaporation were replenished by adding corresponding amounts of MilliQ water.

Preparation and SEM imaging of soil-incubated PBAT films

After 6 weeks of incubation in soil, we carefully removed the silicon wafers carrying the PBAT films from the soils. To chemically fix the cells attached to the surfaces of the PBAT films, we directly transferred the films into a freshly prepared fixation solution (pH 7.4) containing glutaraldehyde (2.5%), sodium cacodylate (0.1 M), sodium chloride (0.1 M), potassium chloride (3 mM), and sodium phosphate (0.1 M). The films were exposed to this solution for 20 min at 25°C and subsequently transferred to a solution of OsO4 in MilliQ water (1%) for 30 min of exposure on ice. Finally, we dehydrated the films in a series of water/ethanol solutions of increasing concentrations (70%, 5 min; 95%, 15 min; 100%, 2 × 20 min), followed by critical point drying of the samples with liquid CO2 (Baltec CPD 030). Critical point drying resulted in detachment of the PBAT films from the wafer. To reattach the films to the wafers for further analyses, we used a double-sided adhesive, electrically conducting carbon tape (Ted Pella, product no. 16084-1). Directly after mounting the films onto the wafers with carbon tape, thin films of platinum (thickness, 10 nm) were deposited on the samples using a sputter coater (Baltec SCD 500). SEM was conducted on a Zeiss Supra 50 VP. Imaging was performed with a secondary electron detector at a working distance of 4.0 mm and an electron high tension of 5.0 kV. These films were also used for NanoSIMS analysis (see below).

PBAT films from the second set, for which wafers were pretreated with Vectabond before solvent casting of PBAT (see above), also detached from the wafers. We decided to reject further analysis of these films (that is, SEM and NanoSIMS).

PBAT film imaging by NanoSIMS

NanoSIMS measurements were performed on a NanoSIMS NS50L (Cameca) at the Large-Instrument Facility for Advanced Isotope Research (University of Vienna). Before data acquisition, analysis areas were presputtered by scanning of a high-intensity, slightly defocused Cs+ ion beam (beam current, 400 pA; spot size, approximately 2 μm). To avoid crater edge effects, scanning during presputtering was conducted over square-sized areas with an edge length exceeding the frame size of the subsequently recorded images by at least 15 μm. Every data set acquired on the soil-incubated polymer films contains image data recorded from (at least) two distinct depth levels, accessed by sequential presputtering with Cs+ ion fluences of 5.0 × 1016 and 2.0 × 1017 ions/cm2, respectively. Application of the lower ion dose density enabled sampling of all cells within the analysis areas, irrespective of their size and/or morphology, whereas the extended presputtering allowed us to gain insight into cellular features contained within the lumen of bulky cells such as fungal hyphae (see, for example, Fig. 4).

Imaging was conducted by sequential scanning of a finely focused Cs+ primary ion beam (2-pA beam current) over areas ranging from 45 × 45 μm2 to 70 × 70 μm2 at a physical resolution of approximately 70 nm (that is, probe size) and an image resolution of 512 × 512 pixels. If not stated otherwise, images were acquired as multilayer stacks with a per-pixel dwell time of 1.5 ms per cycle. 12C, 13C, 12C12C, 12C13C, 12C14N, 31P, and 32S secondary ions as well as secondary electrons were simultaneously detected, and the mass spectrometer was tuned for achieving a mass resolving power of >9.000 (according to Cameca’s definition) for detection of C2 and CN secondary ions. Image data were analyzed with the ImageJ plugin OpenMIMS, developed by the Center for NanoImaging (27). Secondary ion signal intensities were corrected for detector dead time (44 ns) and quasi-simultaneous arrival (QSA) of secondary ions. Both corrections were performed on a per-pixel basis. QSA sensitivity factors (“beta values”) were obtained from measurements on dried yeast cells, yielding 1.1, 1.06, and 1.05 for C, C2, and CN secondary ions, respectively. Before stack accumulation, images were corrected for positional variations originating from primary ion beam and/or sample stage drift. ROIs were manually defined on the basis of 12C14N secondary ion signal intensity distribution images and cross-checked by the topographical/morphological appearance indicated in the simultaneously recorded secondary electron images (see fig. S10). While each cell from unicellular organisms was assigned to an individual ROI, image regions within the polyester surfaces and hyphae were segmented into multiple ROIs. Throughout the article, the carbon isotope composition is displayed as the 13C/(12C + 13C) isotope fraction, given in at%, calculated from the C and C2 secondary ion signal intensities via 13C/(12C + 13C) and 13C12C/(2⋅12C12C + 13C12C), respectively. Owing to superior counting statistics, all carbon isotope composition data shown in the article were inferred from C2signal intensities. We note that we did not observe any significant differences between 13C content values inferred from C2 signal intensities versus C signal intensities. For the line scan analyses displayed in Fig. 4, C2 normalized C14N signal intensities were used as an indicator of the relative nitrogen content {calculated via [12C14N (1 + 13C/12C)]/[12C13C + 12C2 (1 + (13C/12C)2)], whereby the term 13C/12C refers to the 13C-to-12C isotope ratio, calculated from the C2 signal intensities via 13C12C/(2⋅12C12C)}. This quantity formally refers to the relative nitrogen-to-carbon elemental ratio and was used in favor of the relative nitrogen concentration, which is inferable from C normalized C14N signal intensities, to minimize artifacts arising from the considerable topography within the areas of the fungal hyphae (28).


Supplementary material for this article is available at http://advances.sciencemag.org/cgi/content/full/4/7/eaas9024/DC1

Supplementary Materials and Methods

Fig. S1. Mineralization of PBAT films.

Fig. S2. NMR analysis of enzymatic hydrolysis products of PBAT films I.

Fig. S3. NMR spectra of terephthalate, adipate, and butanediol.

Fig. S4. NMR analysis of enzymatic hydrolysis products of PBAT films II.

Fig. S5. NMR analysis of enzymatic hydrolysis products of PBAT films III.

Fig. S6. NMR analysis of enzymatic hydrolysis products of PBAT films IV.

Fig. S7. Mineralization of terephthalate, adipate, and butanediol.

Fig. S8. NanoSIMS analysis of PBAT films after soil incubation I.

Fig. S9. Control experiment for NanoSIMS analysis I.

Fig. S10. Definition of ROIs.

Fig. S11. Control experiment for NanoSIMS analysis II.

Fig. S12. NanoSIMS analysis of PBAT films after soil incubation II.

Table S1. Soil characterization.

Table S2. Characterization of PBAT variants.

Supplementary Appendix. Calculations of the carryover during NanoSIMS measurements.

References (2933)

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

Acknowledgments: We thank S. Probst, M. Jaggi, and F. Strasser for their help with growing E. coli, performing IRMS measurements, and NanoSIMS control sample preparation and data analysis, respectively. Funding: M.T.Z., T.F.N., R.B., H.-P.E.K., K.M., and M.S. thank the Joint Research Network on Advanced Materials and Systems of BASF SE and ETH Zürich for scientific and financial support. M.W. and A.S. were supported by the European Research Council Advanced Grant project NITRICARE 294343. D.W. was supported by the European Research Council Starting Grant project DormantMicrobes 636928. SEM imaging was performed at the Center for Microscopy, University of Zurich. Author contributions: M.T.Z., A.S., D.W., H.-P.E.K., K.M., and M.S. designed the study. M.T.Z., A.S., T.F.N., and R.B. performed experiments. All authors contributed to the writing of the manuscript. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate our conclusions are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.

  • Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).




   Is bamboo the answer to citrus greening in Florida

At the present time, USA production of bamboo products provides but a tiny fraction of the national and international demand which is growing by geometric rates. By eliminating export requirements and import costs the domestic production of bamboo products is on a course to become a major force in Florida agriculture. Bamboo will become the next cash crop for Florida and beyond.  In a collaboration with an Italian bamboo company and Florikan, a producer of NASA chosen controlled release fertilizer we have been able to refine and accelerate  the production, propagation and growth of certain bamboo  species. Due to the elegance of design and safety and efficiency of this controlled release fertilizer it is adaptable to the growth characteristics of bamboo. Application is necessary only once or twice per year or per growth cycle. Edible bamboo shoots in large volume can be achieved in 3 years with mature trunks available for harvest in 5 years.  The future for Florida citrus farmers need not be bleak as they will be the logical recipients of this technology as the demand for bamboo products increases and as Florida farmers and growers realize the vast potential of this “perfect” plant. Just as the original settlers in Florida recognized a natural resource and improved upon it to create a major industry, the new pioneers are poised to begin a new frontier for Florida agriculture. 

Barrett Demler and James Demler Sweetgrass Farms, Sarasota, Florida, USA Published in Fruit Juice Focus Stefan@fruitjuicefocus.com 

Stefan Worsley e contact@juicemarket.info
t +44 (0) 7711 564 219


Continue Reading »

Organic molecules on Mars
On 6 August 2012, the Sample Analysis at Mars (SAM) instrument suite (1) arrived on Mars onboard the Curiosity rover. SAM’s main aim was to search for organic molecules on the martian surface. On page 1096 of this issue, Eigenbrode et al. (2) report SAM data that provide conclusive evidence for the presence of organic compounds—thiophenic, aromatic, and aliphatic compounds—in drill samples from Mars’ Gale crater. In a related paper on page 1093, Webster et al. (3) report a strong seasonal variation in atmospheric methane, the simplest organic molecule, in the martian atmosphere. Both these finding are breakthroughs in astrobiology.
Since NASA’s Curiosity rover landed on Mars in 2012, it has sifted samples of soil and ground-up rock for signs of organic molecules—the complex carbon chains that on Earth form the building blocks of life. Past detections have been so faint that they could be just contamination. Now, samples taken from two different drill sites on an ancient lakebed have yielded complex organic macromolecules that look strikingly similar to kerogen, the goopy fossilized building blocks of oil and gas on Earth. At a few dozen parts per million, the detected levels are 100 times higher than previous finds, but scientists still cannot say whether they have origins in biology or geology. The discovery positions scientists to begin searching for direct evidence of past life on Mars and bolsters the case for returning rock samples from the planet, an effort that begins with the Mars 2020 rover.


NASA Statement on Possible Subsurface Lake near Martian South Pole

Scientists have found evidence of a ‘lake’ on Mars
New evidence shows the presence of a large body of water, almost like a “lake,” one mile beneath the icy surface of Mars, according to a study published in the journal Science. They used data from Mars Express. If confirmed, this would be the largest body of liquid water found on Mars so far.Photo via @cnni
News from Science
Far beneath the ice cap at Mars’s south pole lies a lake of liquid water—the first to be found on the Red Planet:
The view of Mars shown here was assembled from MOC daily global images obtained on May 12, 2003.
Credits: NASA/JPL/Malin Space Science Systems

A new paper published in Science this week suggests that liquid water may be sitting under a layer of ice at Mars’ south pole.

The finding is based on data from the European Mars Express spacecraft, obtained by a radar instrument called MARSIS (Mars Advanced Radar for Subsurface and Ionosphere Sounding). The Italian Space Agency (ASI) led the development of the MARSIS radar. NASA provided half of the instrument, with management of the U.S. portion led by the agency’s Jet Propulsion Laboratory in Pasadena, California.

The paper, authored by the Italian MARSIS team, outlines how a “bright spot” was detected in radar signals about 1 mile (about 1.5 kilometers) below the surface of the ice cap in the Planum Australe region. This strong radar reflection was interpreted by the study’s authors as liquid water — one of the most important ingredients for life in the Universe.

“The bright spot seen in the MARSIS data is an unusual feature and extremely intriguing,” said Jim Green, NASA’s chief scientist. “It definitely warrants further study. Additional lines of evidence should be pursued to test the interpretation.”

“We hope to use other instruments to study it further in the future,” Green added.

One of those instruments will be on Mars later this year. NASA’s InSight lander will include a heat probe that will burrow down as far as 16 feet (5 meters) below the Martian surface. The probe, built by the German Aerospace Center (DLR), will provide crucial data on how much heat escapes the planet and where liquid water could exist near its surface.

“Follow the Water” has been one of the major goals of NASA’s Mars program. Water is currently driving NASA’s exploration into the outer solar system, where ocean worlds — like Jupiter’s moon Europa and Saturn’s moon Enceladus — hold the potential to support life. Even protoplanets like Ceres may explain how water is stored in rocky “buckets” that transport water across the solar system.

 Source : Inge Loes ten Kate Department of Earth Sciences, Utrecht University, Utrecht, Netherlands: i.l.tenkate@uu.nl in Science  08 Jun 2018: Vol. 360, Issue 6393, pp. 1068-1069
DOI: 10.1126/ science.aat2662
Published By:


Cancer is a collection of related diseases involving the abnormal growth of cells within the body.When malignant, cancerous cells divide and spread into surrounding tissues — sometimes at an alarming rate. Many types of cancer result in tumors, which are solid masses of tissue.  Cannabis Vs. Cancer Article By Gooey Rabinski

In the United States, cancer claims the lives of nearly 600,000 people each year and is a $150 billion business. In 2015 alone, it is estimated that more than 1.6 million new cases of cancer will be diagnosed in the United States. Globally, there were 14 million new cases of cancer in 2012 and more than eight million deaths.

According to the National Cancer Institute (NCI), “Studies in mice and rats have shown that cannabinoids may inhibit tumor growth by causing cell death, blocking cell growth, and blocking the development of blood vessels needed by tumors to grow.”http://medicalcannabisreport.com/cannabis-vs-cancer/
Cannabis Vs. Cancer Gooey Rabinski Whaxy.com

The organization noted that animal studies have revealed that cannabinoids are capable of killing cancer cells while — unlike radiation or chemotherapy — protecting normal cells. Continue Reading »

This is a relatively well researched article. The contamination with the processing chemicals is problematic


AVOID! The Toxic Truth About Stevia

By Jane Barthelemy. Stevia is marketed as a healthy sweetener. But surprising new evidence indicates all stevia sold in grocery stores is highly processed with methyl alcohol or other toxic chemicals. Healthy Solution: Look for natural zero-sugar sweeteners. Is your stevia pure? Or is it mixed with other sweeteners and chemicals? Do you wonder how your stevia is refined? What does “natural” mean, if anything? You deserve to know what’s hidden in your stevia. It is essential for your health to learn how stevia is processed, and decide for yourself. This article strips away the marketing hype, with clear evidence that’s very surprising.

Executive Summary:
Yes, Stevia’s green leaves are naturally sweet. However those white powders and clear drops we find in groceries have very little to do with stevia leaves. They aren’t really stevia at all. They’re an extract that’s been super-refined using toxic chemicals, bleach, and marketed as “healthy”. When you look at the chemical refinement process, stevia is no more natural than Aspartame, Splenda, NutraSweet, Equal, Sweet N Low, etc. Everybody is looking for a non-addictive, healthy sweetener. But beware of any sugar-free sweetener that gives you the illusion of a “free ride”, because you may just be deepening your addiction.

Grow Your Own Stevia!
The best stevia is the kind you can grow yourself. When it’s alive you know it’s REAL! Stevia plants grow beautifully in a pot, a garden, a window box, or on your kitchen window sill. Just pull off a leaf when you need to sweeten something. This way you can be sure it’s organic, and the soil is healthy.

What is Stevia Really? How can I Know if it is Pure?
Stevia, or “stevia rebaudiana” is a plant that originates in Brazil with naturally sweet leaves. The leaves can be dried and powdered into a pure sweetener about 40 times sweeter than sugar. These raw, unprocessed stevia leaves have a strong aftertaste akin to licorice, and taste artificial. Pure unprocessed stevia leaves and green powder are not widely available due to their strong aftertaste. If you live in Santa Fe like I do, buy them bulk at the Coop on Alameda near the almond butter grinder. (Or you can order a pound here from Frontier on Amazon).

In grocery stores, we find an entire shelf of “stevia” in the form of processed liquid drops and white powders – all highly refined chemical extractions from the leaves, in the hopes of reducing the aftertaste. The resulting processed sweeteners are called myriad confusing names such as stevia, stevia extract, pure stevia, Rebaudioside A, Reb A, steviol glycosides, etc, and are anywhere from 2X to 350X sweeter than sugar, depending on the blend with other fillers. As a high-intensity sweetener, a little goes a long way, therefore it is often pre-measured in packets or mixed with other fillers such as GMO Maltodextrin, GMO corn Erythritol, inulin fiber, or even cane sugar. Processing is done with a variety of chemicals, such as, methanol, arsenic, ethanol, acetone, and others.

The resulting artificial sweetener called “Stevia” is toxic and unhealthy.
Don’t be fooled by the name, that seemingly innocent stevia we find in grocery stores is a chemical concoction just like Splenda and Aspartame. In fact, it’s highly probable that you’re buying a blend that’s 99.8% Erythritol, a fermented sweetener made from genetically modified corn, with a pinch of refined stevioside powder. Your “Stevia” can be processed, mixed with chemicals, blended in a hundred ways, and still legally be called simply “stevia”. Refined stevioside is sold under countless brand names such as Sun Crystals, SweetLeaf, Truvia, PureVia, Stevia in the Raw, Pyure, and NuStevia to name a few.

Commercial Stevia is bad news.
Stay away from it. That includes Stevioside and Rebaudioside and all the names. All “stevia” in grocery stores is processed with toxic chemicals. If you’re still going for the story that stevia is natural and comes from Peru, know that 85% of all stevia comes from China. Even the world’s top stevia marketer, international sugar giant Cargill, top food manufacturer in the world with over $102.7 billion in 2016 sales, manufacturer of Truvia and PureVia with Coca-Cola and PepsiCo, has all of its stevia produced in China. It’s a small world when you control the sweeteners every body is addicted to.

Take-Home Message:
If you truly want to be free of sugar addiction, then processed stevia and other artificial sweeteners won’t help you. It’s better to skip all forms of Stevia, Truvia, and the blends listed below. The best solution is to use small doses of sugar-free sweeteners, to slowly detox your system from regular sugars. Check out my two preferred sugar-free sweeteners: Just Like Sugar Table Top, and PureLo LoHan by Swanson.

stevia-sugarQuiz: Which sweetener is “Natural”?
1) Sugar cubes, 2) Processed stevioside powder, or 3) Stevia leaves from your garden?


(Most people don’t want too much information. However if YOU are one of those folks that desire the whole truth, read on. To learn how Stevia leaves are processed into a toxic sweetener, it required a bit of digging. As usual, the devil is in the details. To learn the whole Stevia story, continue reading…)

How is Stevia Processed?
Processed stevia is made with a dangerous chemical refining process hidden from the public and deceptively marketed as “natural”. Manufacturers run into the problem that stevia leaves are extraordinarily resilient. The stevia cell walls are so tough that they resist the usual methods of boiling or centrifuging. Producers aim to to extract the active sweet compound, stevioside, and remove the funny aftertaste. In order to concentrate stevia to 300X concentration, toxic chemicals and artificial chemical enzymes are used, such as methanol, kerosene, alcohol, chlorine, ash, acids, titanium dioxide, arsenic, preservatives, chemical stabilizers, and emulsifiers.

The world’s largest producers of stevia hold patents for undisclosed, proprietary extraction methods. These patents belong to industry giants such as Coca Cola, PureCircle in Malaysia and USA, Cargill – maker of Truvia and PureVia, JustBio – A Canadian Biotech firm, McNeil Nuritionals LLC- maker of Splenda, and Chengdu Waggott Pharmaceutical Company in Sichuan China. That’s quite a line-up! Here are 5 common stevia extraction methods I located in public patent records. They all indicate the use of toxic chemicals, which are difficult or impossible to remove.

One of the more popular methods of producing stevia extract was developed by D. Payzant, U.S. Pat. No. 5,962,678. In summary, sweet stevia glycosides are extracted using methanol, a toxic, colorless, volatile flammable liquid alcohol. This method has been used for decades. The major drawback is that a toxic solvent like methanol is difficult to remove. Trace amounts are harmful to health and not ideal for human consumption.
Another common production method comes from Uenishi Hideaki, Japan Patent 54030199. To extract the sweetness and discard a bitter aftertaste, this method also requires the use of various toxic solvents. The removal of solvents requires energy and time, which are not considered cost-effective.
A third production method developed by R. H. Dobberstein, U.S. Pat. No. 4,361,697, uses several toxic solvents including methanol in a complex multi-step process. The major drawback is still the presence of toxic solvents, and their complete removal is not possible and not considered commercially viable.
Sato Toru, Japan Patent JP57005663 uses a new and improved process to extract sweetness from stevia hydrated in water containing alcohol, with the addition of calcium, iron, or aluminum. These compounds are then removed, passed through an acid-ication exchange resin using toxic solvents such as ethanol, acetone, etc. The major drawbacks here are the removal of water from aqueous extract, and removal of toxic solvents, which is not economical.
US. Pat. No. 4,599,403 by Sampath Kumar uses an improved method that is said to be less dependent on toxic chemicals. The major drawbacks are that the aqueous extract is treated first with an acid and then with base and then treated with toxic solvents like n-butanol, which lower the final yield and must ultimately be removed. Again, removal of solvents is not commercially viable, therefore most stevioside products generally contain these toxins.
What’s Really in Your Stevia Bottle?
Well, you can start with the knowledge that there’s almost NO pure stevia out there, except for that rare green powder with a funny aftertaste. (I don’t mind the aftertaste, but many people don’t care for it.) If you want to know what’s really in your stevia, you can try reading the label. However that’s a problem since labels don’t have to disclose all ingredients. Your next hint is serving size. A low serving size of one gram or less is a good indication that the manufacturer is taking full advantage of the legal loophole, and omitting certain chemicals or ingredients. Here’s the loophole: By law, any item under 0.5 grams per serving is not required to be disclosed. So there’s no way you can know for sure what’s really in there. If your Stevia is any of the popular products below, I’ve done some of your homework for you, by reading the labels. However what’s undisclosed we’ll never know.

Popular Stevia Products and their Surprising Ingredients!

1 Better Stevia liquid This is a NOW Foods blend of refined Stevioside with Vegetable Glycerin, a non-glycemic fermented sweetener. 1 tsp liquid = 1 cup sugar sweetness. See Stevia Glycerite.
2 Better Stevia packets NOW Foods makes this product of powdered refined stevioside blended with Non-GMO Rice Maltodextrin.
3 Generic Stevioside Drops See Stevioside Liquid Extract. Generic refined stevioside drops are sold in every grocery chain under their private label, such as Trader Joe’s, Kroger’s, Safeway, Albertson’s, and many other store labels.
3 Generic Stevioside Powder See Stevioside Powder, refined. Refined stevioside powders are sold in grocery chains under their private label, such as Trader Joe’s, Kroger’s, Safeway, Albertson’s, and many other store labels.
4 Generic Stevioside, Industrial See Stevioside Powder, refined. This is a generic powder made of refined stevioside, that is sold on the industrial level as a food additive for the food industry. It is used in a wide variety of food products, such as Good Earth Teas, Celestial Seasonings Tea, Energy Drinks, Sodas, Chocolates, Ice Creams, and Energy Bars. It often contains toxic chemicals, however the amounts are usually under the 0.5 grams per serving, therefore disclosure is not required.
5 Green Leaf Stevia This is a proprietary blend by Swanson made of refined Stevioside powder and high-glycemic non-GMO rice Maltodextrin.
6 Green Stevia Powder This is the pure stuff, and the only healthy stevia. Pure dried stevia leaf is available in a fine green powder that is 30 – 40 times sweeter than sugar. It is raw, and has a peculiar aftertaste. I buy it here.
7 NuNaturals MoreFiber Stevia Baking Blend This is a sugar substitute blend of high glycemic GMO Corn Maltodextrin with refined stevioside. Prepare for a spike in your blood sugar.
8 NuStevia This sugar substitute blends high glycemic GMO Corn Maltodextrin with refined stevioside. Another blood-sugar spike here.
9 PureVia™ Made by Cargill, this sweetener blends genetically modified corn Erythritol with refined Stevioside or Rebaudioside. The Stevia is extracted by proprietary methods we can’t know. There’s nothing natural here.
10 Pyure Organic Stevia A sweetener made from refined stevioside sold in sachets or liquid. It contains agave inulin, refined Stevioside extract, and other unknown ingredients.
11 Rebiana Rebiana is a zero-calorie sweetener produced by proprietary methods by extracting sweetness from the stevia leaf with chemicals and heat, and refining into a high intensity powder that is 200 – 300 times as sweet as sugar. See Stevioside.
12 Rebaudioside Refined Rebaudioside is made from the stevia leaf, where its sweetness is isolated and concentrated using heat and chemicals into a powder about 300X sweeter than table sugar, with somewhat reduced aftertaste. It can be purchased as a white powder or liquid drops. China is the world’s primary producer of rebaudioside. Nothing natural here.
13 Slimstevia A Chinese sweetener similar to Truvia made from genetically modified corn Erythritol with refined Stevioside and/or Rebaudioside. Not natural.
14 Slimtevia This high-intensity sweetener is 3 times sweeter than sugar. It is said to contain high-sugar Fructose, Inulin fiber, FOS (Fructo-oligosaccharides), stevia, and Magnesium Carbonate. This won’t help anyone end the sugar habit.
15 Stevia by Xymogen A sweeter blend of high-glycemic Maltodextrin and refined Stevioside Extract (Rebiana). Prepare for a blood sugar jolt.
16 Stevia dried leaf This is the pure stuff. Unrefined, dried leaves of the South American plant Stevia Rebaudiana are 30–45 times as sweet as table sugar. You can keep this as a potted plant, in bulk dried leaves, or as a green powder. This is a 100% safe sweetener, truly natural (and Paleo). However many people find it has a strong aftertaste. Find it as leaf particles or green powder in food coops and online.
17 Stevia in the raw™ This is a high-glycemic combination of GMO corn Maltodextrin or Dextrose plus refined stevioside. It’s an attractive name, but neither natural nor healthy. Prepare for blood sugar blues.
18 Stevia FOS Blend This is a brand of refined stevioside powder blended with Inulin Fructo-oligosaccharides. It is a zero-calorie, zero carb, sweetener.
19 Stevia Glycerate Proprietary liquid drops produced by NOW Foods, made from refined stevioside and non-glycemic Vegetable Glycerin, a fermented liquid sweetener from oils. 1 tsp Stevia Glycerate = 1 cup sugar sweetness.
20 Steviacane™ This is a blend of refined stevioside with high-glycemic cane sugar by Imperial Sugar Company. Expect a blood sugar jolt here.
21 SteviaClear Drops This is refined stevioside powder in a liquid alcohol solution. The drops are 250 – 300 times as sweet as sugar. Nothing natural here. I suggest first having it tested for methanol and other toxins.
22 Stevioside Liquid Extract These stevioside drops are made from stevia leaves that are refined using methanol and then dissolved in a liquid alcohol solution. There are many sources for stevioside drops, and countless private labels. Most refined Stevioside drops are mixed with other ingredients. The pure stevioside drops are 250 – 300 times as sweet as sugar.
23 Stevioside Powder, refined Refined Stevioside and Rebaudioside are made from the stevia leaf. Its sweetness is isolated and concentrated using heat and chemicals into a powder c. 300 times sweeter than sugar, with reduced aftertaste. China is the world’s primary producer of stevioside. Refined Stevioside and Rebaudioside are often sold in proprietary blends with cane sugar, artificial sweeteners, or other chemicals and rebranded under the generic name of ”Stevia”.
24 Stevita Spoonables A blend of Erythritol and refined Stevioside. Don’t know if it is GMO or NON-GMO corn Erythritol.
25 Steviva Blend A blend of high quality Non-GMO Erythritol with refined Stevioside powder. Steviva Blend is twice as sweet as sugar. There’s nothing natural here.
26 Sun Crystals® A blend of cane sugar mixed with refined stevioside. Prepare for sugar shock.
27 Sweet Serum A low-carb, low-glycemic liquid sweetener that contains organic raw agave inulin, Yacon root and Stevioside. Sweet Serum has a concentrated sweet honey taste. 1 teaspoon Sweet Serum is equal in sweetness to about 5 teaspoons sugar. Nothing natural here.
28 Sweet Simplicity® A Sugar Substitute made from genetically modified corn Erythritol, Fructose sugars and Natural Flavors by Whole Earth Sweetener Company, the makers of PureVia. Prepare for insulin shock.
29 Sweet’nVit stevia A high intensity sweetener developed by the European firm Vitiva containing refined Stevioside, genetically modified Corn Erythritol and Maltitol, a fermented sweetener.
30 SweetLeaf Stevia Shaker A blend of refined stevioside powder and inulin. Nothing natural here.
31 Truvia™ A blend of GMO corn Erythritol, refined Rebaudioside, and other ingredients by Cargill.
32 ZSweet® A sweetener that can be used cup for cup like sugar, made from Non-GMO Erythritol and highly refined Stevioside or Rebaudioside.

Stevia was once a simple plant used by the Guarani Indians in South America for healing. But our world-wide craving for sweetness, along with modern food processing methods have changed all that. Now stevia is refined with toxic chemicals in private proprietary procedures deeply linked to the largest international corporations and the sugar industry. Most of our stevia is produced in China, and then marketed as our most beloved natural sweetener. If you still believe your stevia to be healthy, check out the links below for a journey of deception and international intrigue that will make your hair stand up on end.


Patent – Manufacturing method of pure natural high-purity stevioside – CN 102199177 (Translated from Chinese) http://www.google.com/patents/CN102199177A?cl=en

Patent – High-purity rebaudioside A and method of extracting same https://www.google.com/patents/US7923541

Patent – Process For Extraction And Debitterizing Sweet Compounds From Stevia Plants http://www.freepatentsonline.com/y2016/0015066.html

Patent – Process for production of steviosides from stevia rebaudiana bertoni – US 20060142555 A1 http://www.google.com.ar/patents/US20060142555

Method for extracting active ingredient of natural product (stevia) and uses thereof CN 101138686 (Translated from Chinese) A http://www.google.com/patents/CN101138686A?cl=en

The Aspartame / NutraSweet Fiasco http://www.stevia.net/aspartame.htm

How the Feds Set Frankenstein Free on the Farm, by Dr. Al Sears, M.D. http://www.bibliotecapleyades.net/ciencia/ciencia_geneticfood140.htm

Is Stevia Paleo? https://www.primalorganicmiami.com/is-stevia-paleo/

Cargill to Settle Deceptive Marketing Lawsuit alleging Truvia, Stevia Based Sweetener is Not Natural. http://www.foodnavigator-usa.com/Regulation/Cargill-to-settle-deceptive-marketing-lawsuit-alleging-Truvia-stevia-based-sweetener-is-not-natural

Don’t confuse consumers with stevia messages, by Russ Bianchi

A Tale of Two Sweeteners – Stevia and Aspartame

Stevia Leaf – Too Good To Be Legal?

Stevia – A Natural Choice, by Dr. Betty Martini


It’s been a while, but we have received the final finished footage of our company Florikan and Chairman Ed Rosenthal and President Eric Rosenthal induction into the Space Foundation‘s Space Technology Hall of Fame. And our first thought was share it with our fans! Enjoy!

Quite the honor !