Patent Application
20220400619
The following invention is relevant to the field of agronomy. It specifically relates to a mobile light exposure device for improving the yield and quality of biological, particularly plant, material, as well as to the process and associated uses.
Improving crop yields and the quality of the biological material produced is an important current issue in the development of sustainable and environmentally friendly agriculture.
Even today, the use of phytopharmaceutical products remains the most effective solution for optimising crop yields.
Phytopharmaceutical products are any substances or formulations used to prevent, destroy or repel organisms undesirable to agriculture or public health, to stimulate plant metabolism, growth or morphogenesis. The most commonly used types of phytopharmaceutical products are pesticides, hormones and growth enhancers. A phytopharmaceutical product often takes the form of a chemical substance with or without additives.
The use of these substances is often recommended for large-scale cultivation such as in fields or greenhouses, as well as in public green areas and private gardens. As it is, crops are more difficult to control due to the presence of many biotic and abiotic factors that can affect the growth and, in the case of agricultural production, the final quality of the plants at harvest.
The harmful effects of phytopharmaceutical products on the environment and the contamination of drinking water resources such as groundwater have been taken into consideration.
Many animal species such as birds or non-pest insects (e.g. pollinators) are affected by phytopharmaceutical products. A direct poisoning can occur due to the presence of residues of phytopharmaceutical products, especially when animals enter treated fields.
Runoff from phytopharmaceutical products can also affect aquatic fauna, leading to the death or reduction of fish and amphibian populations. This fatality can be direct or indirect. Repeated exposure to phytopharmaceutical products can cause physiological and behavioural changes, but can also lead to decomposition of the plants, thus depriving fish of oxygen.
The consumption of products or by-products from agriculture using phytopharmaceutical products is the subject of numerous studies to determine the effects on health in the medium and long term.
The WHO also warns of the direct and indirect hazards to humans from the use of and exposure to phytopharmaceutical products, which can manifest themselves as acute poisoning or chronic side-effects.
Exposure to phytopharmaceutical products is also suspected of being the cause of numerous disorders and pathologies such as cognitive disorders; anxiety-depressive disorders; disorders of child development during pregnancy; fertility problems; problems of metabolisation of phytopharmaceutical products; metabolic problems (obesity, type II diabetes, thyroid dysfunction, . . . ); various cancers such as leukaemia, testicular cancer, brain tumours, melanoma, Hodgkin's disease, Kahler's disease; Parkinson's disease; Alzheimer's disease; and amyotrophic lateral sclerosis.
In view of the above, there is now a need to develop and optimise alternative techniques that achieve results that are substantially as effective as with the conventional use of phytopharmaceutical products but less harmful to health and the environment. Therefore, the Applicant was interested in plant treatments without the use of phytopharmaceutical products. Among these treatments, the treatment of crops with light appears to be an interesting and promising avenue.
Light treatment of crops is an interesting alternative to overcome these problems and does not have the negative environmental and health effects of phytopharmaceutical products described above.
Light is one of the most important environmental factors regulating plant growth and development. Plants need light not only for photosynthesis, but also for precise regulation of their development.
Several abiotic factors can influence plant metabolism and vigour, such as lack of water, excessive temperatures or ultraviolet radiation.
Ultraviolet (UV) radiation, also known as “black light” because it is not visible to the naked eye, is electromagnetic radiation of shorter wavelength than visible light but longer than X-rays. They can only be observed indirectly, either by fluorescence or with the help of specialised detectors.
Ultraviolet, blue and red radiation are involved in various photomorphogenic responses. UV-B wavelengths (280-315 nm) are biologically active and when applied at non-harmful doses induce changes in gene expression, physiology, metabolite accumulation and plant morphology.
As with other components of the solar spectrum, the effects of UV-B on plant physiology are influenced by hormones. Some of these hormones, such as abscisic acid, jasmonic acid, salicylic acid and ethylene, appear to be mainly factors of adaptation to environmental variations and stress. Other hormonal pathways using gibberellins and auxins in particular confer mainly morphological changes.
The use of UV-B radiation therefore influences the metabolism of plants to act on their morphology and in particular allows the synthesis of molecules such as flavonoids or the accumulation of phenolic pigments such as anthocyanins.
However, there are practical problems when using UV-B. Prolonged exposure of several hours or even days is required to be effective. However, prolonged exposure is difficult to implement on crops outside greenhouses.
UV-C radiation, on the other hand, is sufficiently energetic to break chemical bonds. Absorption at particular wavelengths can be associated with resonance effects in which certain energy levels in an atom or molecule are almost equalled by the energies of incident photons. Used in high doses, UV-C can therefore cause direct damage to various molecules (nucleic acids, proteins, etc.).
With this in mind, UV-C radiation is commonly used for surface disinfection purposes.
Once the cuticular barrier is crossed in plants, UV-C radiation is more strongly absorbed than UV-B or UV-A. This is due to the fact that UV-C radiation excites many more molecules than UV-B and UV-A radiation, which are only absorbed by highly complex compounds or those with C=O or C+N functions, for example. Since it is absorbed more strongly than UV-B and UV-A, UV-C radiation is much less likely to penetrate deep into the tissue and thus cause damage, particularly on DNA molecules.
As UV-C is safer for the treated plants than UV-B or UV-A, it has been tested on different types of plant organs with the idea of identifying hormetic doses, i.e. doses that produce positive biological effects.
Earlier patent documents disclose the use of UV, in particular UV-C, on plants, but for different purposes than the invention in question.
Document WO9533374 details the use of a laterally moving rail vehicle to emit a glow discharge consisting of a mixture of UV-A, B, C, visible and infrared light. The exposure time of the plants to this light discharge is less than ten seconds, preferably three seconds. The use of this device is intended for the destruction of unwanted plants. As such, this paper discourages the use of radiation, particularly UV-C, to improve the yield and quality of biological material.
Document WO2007/049962 describes the use of a transport means for conveying plants under a light source consisting of at least 90% UV-C. This device is used for plant or fungus treatment purposes to control the growth of pathogens, insect pests and also for the destruction of aerial plant parts. As such, this paper does not consider the use of light pulses to improve the yield and quality of biological material.
Document FR3063974 describes a device with a conveyor belt for direct exposure of harvested plants to UV-C light pulses to decontaminate their surface which is contaminated with phytopharmaceutical products. However, this document does not describe a mobile device comprising locomotion means for moving the device, nor does it describe the use of the device to improve the yield and quality of biological material.
Finally, document DE19900616A1 describes a method for stimulating the production of anthocyanins in plants and fruits using a device emitting light at wavelengths in the visible spectrum but also UV-A and UV-B. However, the method described does not mention the use of UV-C for the improvement of the yield and quality of biological material, nor an exposure time of the order of seconds.
Taking the above into account, one problem that this invention proposes to solve consists in developing an alternative device that makes it possible to dispense with, or at least greatly limit, the use of phytopharmaceutical products. This alternative must ensure sufficient yield and quality of biological material.
Moreover, this device must be more ecological in order to preserve the environment, but also simple to use.
Its applications must be feasible at different scales, both in the greenhouse and in the field.
The first goal of the solution to this problem is a mobile light exposure device for improving the yield and quality of biological material comprising:
In particular, the Applicant has been able to demonstrate that UV-C radiation is less harmful than UV-B or UV-A radiation in plants. UV-C radiation is less penetrative to the tissue due to the micro-relief of epicuticular wax crystals that cause it to scatter on the leaf surface. After crossing the epicuticular barrier, penetration is all the lower as there is a high molecular uptake in the plant cells under the cuticle.
With this in mind, the idea of using the more energetic UV-C instead of UV-B for much shorter periods of time therefore appears to be more advantageous to the Applicant.
A second objective for the invention is a process for improving the yield and quality of biological materials comprising the following steps:
Finally, the third objective is the use of a device according to the invention for modifying the physiology of a biological material.
The invention developed consists of a mobile light pulse emitting device with qualities of adaptability and ease of regular use for the treatment of surfaces of variable size.
The applicant has been able to demonstrate that UV-B induced changes include:
These same changes can be observed using UV-C radiation with shorter exposure times when used at a given power and do not a priori cause negative effects on plants. They are perceived by plants and stimulate signalling and metabolic pathways influencing growth, development, branching, flowering, yield and quality under both normal and stress conditions. Applied at hormetic doses, i.e., doses that allow positive biological effects to be obtained, without negative side effects, they can be used in the field of agronomy to improve the yield and quality of biological material in order to obtain improved commercial products, particularly in terms of their concentrations of compounds of interest.
The invention and the advantages thereof will be better understood on reading the following description and non-limiting embodiments, illustrated with reference to the annexed drawings in which:
In this description, unless otherwise specified, it is understood that when an interval is given, it includes the upper and lower bounds of the interval.
According to the invention, the mobile light exposure device 1 for improving the yield and quality of biological material 2 as illustrated in
By light exposure, the applicant means one or more light sources 9 from said device emitting at wavelengths between 200 nm and 700 nm.
The first module of the mobile light exposure device 1, comprising one or more discharge lamps according to the invention, allows one or more light pulses to be emitted. Non-limiting examples of lamps that can be used are low, medium or high pressure lamps, pulsed light or Xenon lamps, Excimer lamps, LED lamps or mercury vapour lamps (254 nm).
The light pulses are characterised in particular by their duration and wavelength.
The durations of the light pulses are necessarily less than two seconds, preferably less than or equal to one second.
Preferably, the duration of the light pulses is between one second and one thousandth of a second. Preferably, it is between one second and one hundredth of a second. Particularly preferred values used by the Applicant are one second, one tenth of a second, one hundredth of a second or 300 μs or 500 μs.
The number and frequency of the light pulses are modulated according to the nature of the biological material to be treated 2.
The wavelengths of the light pulses are generally between 200 nm and 700 nm (UV-C, UV-B, UV-A, visible light), preferably between 200 and 280 nm (UV-C). Even more advantageously, they are between 250 and 265 nm.
Even more preferably, the light pulses can be:
The device according to the invention allows the improvement of the yield and quality of the biological material 2.
Biological material 2 means plant material, fungus or microorganisms or media from the culture of microorganisms. Preferably, the biological material 2 is a plant material. Plant material is a whole plant or part of a plant such as a cell, tissue, leaf, fruit, stem, flower or root.
Preferably, said plant material comes from farms comprising plantations. These plantations are derived from vitro-plant, agriculture, forestry or horticulture such as vegetable, fruit, cereal, oilseed or protein crops.
As non-limiting examples of usable plant material, the following plant families can be mentioned: Amaranthaceae, Apiaceae, Arecaceae, Asteraceae, Brassicaceae, Cannabaceae, Cucurbitaceae, Fabaceae, Liliaceae, Musaceae, Poaceae, Rosaceae, Rubiaceae, Rutaceae, Solanaceae and Vitaceae.
Another example is grass, i.e., any annual or perennial, non-woody plant belonging to the Angiosperms (monocotyledons or dicotyledons), generally green in colour. More specifically, grass commonly refers to the grasses, especially the forage grasses, which make up the grasses, meadows and lawns, and the morphologically related families rushes and sedges.
Preferably, the plant species used are: Allium ampeloprasum var. porrum (Leek), Allium cepa (Onion), Allium sativum (Garlic), Brassica oleracea (Cabbage) including var. italica (Broccoli) and var. botrytis (Cauliflower), Cannabis sativa (Cannabis), Capsicum annuum (Pepper), Cucurbita pepo (Courgette), Cucurbita maxima (Pumpkin), Daucus carota (Carrot), Elaeis guineensis (Oil palm), Fragaria x ananassa (Strawberry), Glycine max (Soya), Helianthus annuus (Sunflower), Hordeum vulgare (Barley), Lactuca sativa (Lettuce), Malta domestica (Apple), Mangifera indica (Mango), Musa spp. (Banana), Nicotiana tabacum (Tobacco), Oryza sativa (Rice), Prunus persica (Peach), Prunus avium (Cherry), Pyrus communis (Pear), Raphanus sativus (Radish), Rosa hybrida (Rose), Secale (Rye), Solanum lycopersicum (Tomato), Solanum tuberosum (Potato), Triticum spp. (Wheat), Vitis vinifera (Vine), Zea mays (Maize).
According to the invention, the device enables one or more light pulses to be emitted from one or more light treatment panels.
These light pulses may be of different wavelength, power and/or duration. Similarly, it is possible to consider superimposing different light pulses (in terms of wavelength, duration or power) during the passage of the device.
In particular, this makes it possible to use different light pulses simultaneously, separately or spread out over time.
The first light pulse emission module 3 of the device according to the invention comprises at least one light treatment panel whose surface is between 0.01 m2 and 10 m2.
Preferably, the area of the light treatment panel is between 0.01 m2 and 5 m2. Even more preferably, the surface area of the panel is between 0.01 m2 and 3 m2. The surface area of the panels preferably used is approximately 0.4 m2, 0.8 m2, 1 m2 or 1.2 m2.
According to the invention, the mobile light exposure device 1 for improving the yield and quality of biological material comprises a second adjustment module 6.
The second adjustment module 6 can be controlled remotely or directly on the device.
Preferably, the second adjustment module 6 allows both adjustment of the optical power density of the treatment panel and the temperature of the panel.
The temperature of the panel is regulated actively (e.g. by a fan) or passively (e.g. by a thermal diffuser) by a temperature control unit 12 which changes the temperature based on data it receives from a temperature sensor 13.
Preferably, the second adjustment module controls a mechanical adjustment module 8 which ensures the correct positioning of the panels with respect to the biological material 2, in particular when the biological material 2 is presented in the form of a low culture as illustrated in
The optical power density of the panel is between 100 W/m2 and 10 000 W/m2, preferably between 300 W/m2 and 3 000 W/m2. The sources used can be discharge lamps (including low, medium or high pressure lamps, pulsed or Xenon light, Excimer lamps) or LEDs. The above-mentioned light sources 10 can be advantageously mounted on a so-called reflector body 9 with reflectors 11 as well as the temperature control unit 12, in order to control the light beam as shown in
Even more preferably, the optical power density of the panel is between 500 W/m2 and 2500 W/m2, preferably between 1000 W/m2 and 2000 W/m2.
Of course, the person skilled in the art will be able to adapt the above-mentioned settings according to the surface and the plant material to be treated.
According to the invention, the mobile light exposure device 1 for improving the yield and quality of biological material 2 also comprises a locomotion means enabling the device 4 to be moved, preferably at a speed of between 1 and 10 km/h. The means of locomotion 4 is advantageously a traction or propulsion means.
Preferably, the speed of the mobile device is between 1 and 10 km/h. Preferably again, it is between 2 and 5 km/h. The particularly preferred speed values used by the Applicant are 4 km/h, 3.6 km/h, 2.5 km/h and 1.8 km/h.
The means of locomotion 4 may or may not include driving wheels that can travel on any type of road or rail. Depending on the nature of the surface, it may refer to a traction device consisting of wheels and assisted or not by motor. Non-limiting examples include but are not limited to:
Preferably, the means of locomotion 4 used is a traction or propulsion device consisting of wheels assisted by a thermal or electric motor.
The size of the areas to be treated is variable and can generally range from 0.001 m2 to 100 hectares. Preferably, the surface area of the areas to be treated corresponds to the size of a crop field, a nursery, a green space, but also to a product obtained in post-harvest.
The device according to the invention is advantageously powered by an independent and/or autonomous energy source 7. Preferably, the power source is in the form of a battery and/or an alternator. Alternatively, the device can also be assisted by the presence of solar panels, an electric, combustion, internal combustion or hybrid engine.
Advantageously, the device is powered by a single-phase or three-phase, 50 or 60 Hz alternator delivering a voltage of between 110 V and 500 V. Preferably, this alternator has a voltage stabilisation device and will produce a power at least equal to the power of the lamps. For example, a device with a 2 m2 panel will need to be powered by an alternator providing between 250 W and 25,000 W depending on the optical power density selected.
The invention also relates to a process for improving the yield and quality of biological materials comprising the following steps:
Preferably, the method according to the invention is adapted to a biological material 2 which is a plant material such as a fruit, vegetable, seed, vitro plant, tuber or any other part of a plant.
Finally, the invention has as a last object the use of a device according to the invention, for the modification of the physiology of a biological material 2.
The light pulses emitted by the use of the invention modify the physiology of the biological material. Modification of the physiology of a biological material 2 means mechanical, physical and/or biochemical modification as a result of its interaction with the environment.
According to a first preferred embodiment, the object of the invention is the use of a device for modifying the primary and/or secondary metabolism of a biological material 2, preferably of a plant material.
Preferably, the effects are observable on the primary and secondary metabolism of plants.
Primary plant metabolism refers to all metabolites directly involved in normal growth, development and reproduction. Secondary metabolism involves all metabolites existing in low concentrations in plants.
Observable effects include a quantitative and qualitative change in the production of chemical molecules produced by the primary and secondary metabolism of plants. By way of non-limiting examples, the molecules concerned may be monosaccharides, glucosinolates, amino acids, proteins, lipids, terpenoids, phenolic compounds, alkaloids or cannabinoids (THC for tetrahydrocannabinol and CBD for cannabidiol).
The Applicant has observed that the process according to the invention allows an increase in the concentration of cannabinoids (THC, CBD), flavonoids and terpenes.
The quantitative and qualitative modification of the production of chemical molecules produced by the primary and secondary metabolism of plants can lead to the modification of the growth and/or development, the increase of the defences as well as the morphogenesis of a biological material 2, preferably of a plant material, especially under stress.
According to a second preferred embodiment, the object of the invention is the use of a device for modifying the growth and/or development of a biological material 2, preferably of a plant material.
According to a third preferred embodiment, the invention relates to the use of a device for increasing the defences of a biological material 2, preferably a plant material.
Surprisingly, the Applicant was also able to demonstrate that the use of the device according to the invention and in particular the application of light pulses to the biological material 2 made it possible to obtain the following effects:
Thus, the results obtained by using this invention may also be applicable in the following fields, as non-limiting examples: pharmacology, cosmetics, the paper industry and its derivatives, the flavour and perfume industry, the “green” chemical industry, the food industry, animal feed, nutraceuticals.
The present invention will now be illustrated by means of the following examples.
The first results obtained with mercury vapour lamps (254 nm) show a superiority of UV-C flashes (one second) compared to conventional exposures (one minute). UV-C flashes show stimulating effects on secondary metabolism, growth, yield and production quality.
The tobacco plants were sown at D0.
Observations were made on the effects of UV-C treatments applied at a dose of 1 kJ/m2, either as one second flashes (D1) or as conventional one minute exposures (D2), on the nicotine concentration of tobacco leaves from plants grown in pots under glass. The trial also included an untreated control.
The device used for the UV-C treatments consisted of mercury vapour lamps (254 nm) and allowed comparison of flashes and conventional exposures at the same dose and wavelength.
Several UV-C treatments were carried out, namely at D47, D54 and D61 and at D68. Harvesting took place one week later at D75.
Two basal leaves were collected per plant, from ten plants per sample. That is a total of 20 leaves collected per sample. n=3. The leaves were oven-dried at about 80° C. for 48 hours and then ground.
Nicotine was determined by HPLC-DAD 260 nm and the concentration expressed on a dry matter basis.
Tab. 1. Effect of conventional and UV-C flash exposures on nicotine concentration in tobacco leaves. Different letters indicate significant differences at the 5% level.
The results in Table 1 show that UV-C flashes substantially increase (+17%) the nicotine concentration of tobacco leaves compared to the control, while conventional exposures decrease it (−8%).
The leek plants were sown at DO.
Observations were made on the effects of UV-C flash treatments applied at three doses, 400, 800 and 1200 J/m2 on the mass and length of the drums. The trial also included an untreated control (UT). The scheme was completely randomised with n=11.
The device used for UV-C treatments consists of mercury vapour lamps (254 nm). Doses of 400, 800 and 1200 J/m2 were obtained with exposure times of 1 s, 2 s and 3 s, respectively.
Four treatments were performed, every 7 days, between D23 and D44.
The drums were harvested at D51, weighed and measured for length.
The results in
From the results in
The red anthocyanin variety GNIFF AB from the Sainte Marthe farm was used in this trial.
Sowing took place at D0 and cultivation was carried out in a greenhouse with temperatures between 17° C. and 26° C.
The device used for the UV-C treatments consisted of mercury vapour lamps (254 nm). The doses of 100, 300 and 500 J/m2 were all obtained with an exposure time of is by adjusting the distance between the lamps and the plants. The experimental set-up also included a control n=35.
The carrots were harvested at D60. The total weight, length and diameter of the roots were measured. The length of the leaves was also measured.
Tab. 2. Effect of different doses of UV-C applied as is flashes on leaf length, total weight, length, diameter and length to diameter ratio of carrot roots. *, ** and *** correspond to differences with the control, significant at the 10%, 5% and 1% thresholds, respectively (Student's t test).
UV-C flashes applied during cultivation increase leaf length and total weight and root diameter. Only the 100 J/m2 treatment increases the root length. It can be seen that UV-C flashes stimulate growth and yield. They can also exert morphogenetic effects and therefore on the quality of presentation by modifying the length/diameter ratio of the roots (treatments at 300 and 500 J/m2). In general, the effects on leaf length, total weight and root diameter are more pronounced the higher the applied rate.
The red anthocyanin variety GNIFF AB from the Sainte Marthe farm was used in this trial.
Sowing took place at D0 and cultivation was carried out in a greenhouse with temperatures between 17° C. and 26° C.
The device used for the UV-C treatments consisted of mercury vapour lamps (254 nm). The doses of 100, 300 and 500 J/m2 were all obtained with an exposure time of is by adjusting the distance between the lamps and the plants. The experimental set-up also included a control n=35.
The root tubers of the carrots were harvested at D60 and ground in liquid nitrogen and the powder was used for the determination of phenolic compounds, flavonoids and all anti-free radical species.
UV-C flashes applied during cultivation increase the amount of phenols (
Refrigerated strawberry plants of the variety ‘Gariguette’ were planted in 2 litre pots in a glasshouse at D0.
The device used for UV-C treatments consists of mercury vapour lamps (254 nm). The treatments consisted of applying a dose of 800 J/m2 (in 1 s) on the following days: D5, D12, D19 and D26. The treatments were carried out on normal plants and on plants treated with a defence elicitor treatment)(Vacciplant®). Vacciplant® treatments were carried out at D10 and D24. In addition to normal plants treated with UV-C and plants treated with UV-C and Vacciplant® , the randomised trial included untreated plants and plants treated only with Vacciplant® n =20.
Flowering was noted on the following days: J10, J12, J14 since the start of the test, noted as J0, J+2, J+4 in the figure).
Strawberry plants of the variety ‘Gariguette’ were planted in 21 pots under glass at D0.
The device used for the UV-C treatments consisted of mercury vapour lamps (254 nm). Observations were made on the effects of two UV-C doses (D1=800 J/m2, D2=1600/m2), obtained after 1 and 2 s of exposure respectively. The UV-C treatments were carried out on the following days: J18, J25, J32 and J45. The trial included a control (TNT) and n=20.
Observations were made at D53 on the number of flowers, the number of green fruits and the number of ripe fruits.
At D53, there was no difference in the number of flowers between the treated plants and the control (results not shown), which is consistent with the end of trial observations presented in Example 4.
Several Cannabis sativa L. plants were planted at D0 in Spain, in 15 L pots filled with fertilised All Mix potting soil, in Mylar ‘Black boxes’. Watering was provided every 3 days. The imposed temperatures were 19° C. at night and 26° C. during the day. During the growth phase (3 weeks), the photoperiod was 18 h day/6 h night with lighting provided by MH 250W 6600° K lamps. During the flowering phase (8 weeks), the photoperiod was 12 h day/12 h night with lighting provided by HPS 400W 2700° K lamps. Ventilation was provided with a flow rate of 15 m3 /h.
The device used for the UV-C treatments consisted of mercury vapour lamps (254 nm). Observations were made on the effects of a single dose of UV-C given as a is flash, first from the 5th week of flowering, and a second time one week later.
Samples were taken one week after the second light treatment from pooled plant material per treatment (2 grams per treatment). The comparison focused on the effect of UV-C flash treatments versus an untreated control.
Tab. 3. Effect of UV-C flash exposures on cannabinoid concentration in Cannabis sativa L. inflorescences expressed as mg/g dry matter.
The results in Table 3 show that UV-C flashes substantially increase (+621%) the THC concentration compared to the control. CBD levels are only trace, as expected.
The trial was conducted on vines located on a ‘Merlot’ plot randomised in 3 blocks. Plants treated with UV-C flashes were exposed to illuminations of about one second every 10 days from April to July. The lamps used for the light treatments were UV-C (254 nm) amalgam lamps mounted on a prototype tractor moving between the crop rows. The treatments consisted of flashes of approximately one second, delivering 800 J/m2/flash.
10-13 randomly selected plants were harvested individually per block (n=35) in September. The values in the table are averages+standard deviations, expressed in kg of clusters per vine. Different letters indicate significant differences at the 5% level (non-parametric Kruskal Wallis test).
| Number | Date | Country | Kind |
|---|---|---|---|
| 1908196 | Jul 2019 | FR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2020/069480 | 7/10/2020 | WO |
