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    • 专利摘要:
    The composition, processes, and soil treated of the present invention relates to compositions comprising chitosan, glucosamine and amino acids, wherein the concentration of said chitosan is greater than 1.5% by weight and said glucosamine is greater than 1.5%. by weight The composition may also include solid chitin, but generally not more than about 2% by weight. The composition may also include trace elements, protein and other polysaccharides. The composition is generally a liquid, but may be a solid. In most embodiments, the solid may be reconstituted with water prior to use. In preferred embodiments, the composition comprises hytd and at least one of, hyta, hytb and hytc. in still other embodiments the composition comprises hytd and two or more of hyta, hytb and hytc. The composition may also comprise hytd, hyta, hytb and hytc. In disclosed processes soil, seed, seedlings or plant foliage are in contact with hytd or any of the above compositions.
    公开号:BR112013033319B1
    申请号:R112013033319-7
    申请日:2012-06-25
    公开日:2019-10-08
    发明作者:Jaime López-Cervantes
    申请人:Agrinos AS；
    IPC主号:
    专利说明:

    COMPOSITION, PROCESSES, AND TREATED SOIL
    This application claims priority for US Provisional Application Serial No. 61 / 500,543 filed on June 23, 2011.
    TECHNICAL FIELD [001] The processes and compositions are publicized, which increase agricultural production, increase plant defense processes, decrease the level of plant pathogens and reduce the amount of fertilizer used.
    BACKGROUND OF THE INVENTION [002] Microbes were previously used in agriculture. Examples include those disclosed in US Patents 4,952,229; 6,232,270 and 5,266,096.
    [003] Chitin has also been used in agriculture as a protein complex (US Patent 4,536,207) or in combination with several microbes (US Patent 6,524,998 and 6,060,429).
    [004] Chitosan in combination with other components has been used in agricultural applications. See, for example, US Patents 6,649,566; 4,812,159; 6,407,040; 5,374,627 and 5,733,851. It was also used to treat seeds from cereal crops. See US Patent 4,978,381. US Patent 6,524,998 also discloses that chitosan can be used in combination with specific microbes for agricultural use.
    [005] HYTb, alone or in combination with HYTc and the microbial composition HYTa are useful in the treatment of soil, seeds, seedlings and foliage, as disclosed in US Patent Application Serial No. 61 / 355,447 filed on June 16, 2010 entitled “Microbial Process and Composition for Agricultural Use” and US Patent Application Serial No. 13 / 160,333 filed on June 14, 2011 entitled “Microbial Process and Composition”, each of which is incorporated into this document by reference in its entirety .
    [006] Notwithstanding the above, there is a need to provide improved compositions and processes that enhance the harvest and reduce the amount of conventional fungicides and insecticides used in agricultural and horticultural applications.
    SUMMARY OF THE INVENTION [007] Compositions are disclosed comprising chitosan, glucosamine and amino acids, where the concentration of chitosan is greater than 1.5% by weight and glucosamine is greater than 1.5% by weight. In the preferred embodiments, the concentration of chitosan is 2 to 2.5% by weight and
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    2/31 of glucosamine is 2 to 6% by weight. The composition can also include solid chitin, but generally no more than about 2% by weight.
    [008] The composition can also include trace elements, protein and other polysaccharides.
    [009] The composition is usually a liquid, but it can be a solid. In most embodiments, the solid can be reconstituted with water before use.
    [010] In preferred embodiments, the composition comprises HYTd and at least one of, HYTa, HYTb and HYTc. In still other embodiments, the composition comprises HYTd and two or more, HYTa, HYTb and HYTc. The composition can also include HYTd, HYTa, HYTb and HYTc.
    [011] In the processes disclosed, soil, seed, seedlings or plant foliage are in contact with HYTd or any of the above compositions.
    [012] The composition of treated soil comprising soil treated with HYTd or any of the above compositions is also disclosed.
    [013] The treated plant comprising the HYTd treated plant or the composition of any of the above compositions is also disclosed.
    [014] Treated seed or seedlings comprising seed or seedlings treated with HYTd or any of the above compositions are also disclosed.
    BRIEF DESCRIPTION OF THE DRAWINGS [015] Figure 1 shows the results of the treatment of asparagus with HYTa + HYTb.
    [016] Figure 2 shows the results of the treatment of asparagus with HYTa + HYTb + HYTd.
    [017] Figure 3 is a graph showing the number of pieces and the size distribution of potatoes treated with HYTa compared to the control.
    [018] Figure 4 is a graph showing the number of pieces and the mass distribution of potatoes treated with HYTa compared to the control.
    [019] Figure 5 contains photographs comparing the potatoes obtained after treatment with HYTa compared to the control.
    [020] Figure 6 is a graph showing the number of pieces and the size distribution of potatoes treated with HYTa compared to the control.
    [021] Figure 7 is a graph showing the number of pieces and the mass distribution of potatoes treated with HYTa, HYTc, HYTc and HYTd compared to HYTa.
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    3/31 [022] Figure 8 contains photographs comparing potatoes obtained after treatment with HYTa, HYTc, HYTc and HYTd compared to HYTa.
    [023] Figure 9 is a graph showing the number of pieces and the size distribution of potatoes treated with HYTa, HYTc, HYTc and HYTd compared to Metam-sodium.
    [024] Figure 10 is a graph showing the number of pieces and the mass distribution of potatoes treated with HYTa, HYTc, HYTc and HYTd compared to Metam-sodium.
    [025] Figure 11 contains photographs comparing potatoes obtained after treatment with HYTa, HYTc, HYTc and HYTd compared to Metamsodium.
    [026] Figure 12 is a flow chart showing the digestion of the crustacean to form HYTb and HYTc. HYTc and HYTb are subsequently processed with HQE to form HYTd, a solution with relatively high amounts of chitosan and glucosamine compared to HYTb.
    [027] Figure 13 is a flow chart showing the digestion of fungi, including filamentous fungi, yeasts and / or insects to form HYTb and HYTc. HYTc and HYTb are optionally further processed with HQE to form HYTd, a solution with relatively high amounts of chitosan and glucosamine compared to HYTb.
    DETAILED DESCRIPTION [028] The compositions are disclosed comprising chitosan, glucosamine and amino acids, where the concentration of said chitosan is greater than 1.5% by weight and said glucosamine is greater than 1.5% by weight. In the preferred embodiments the concentration of chitosan is 2 to 2.5% by weight and that of glucosamine is 2 to 6% by weight. The composition can also include solid chitin, but generally no more than about 2% by weight. The composition can also include trace elements, protein and other polysaccharides. The composition is usually a liquid, but it can be a solid. In most embodiments, the solid can be reconstituted with water before use. In preferred embodiments, the composition comprises HYTd. In other embodiments, the composition comprises HYTd and at least one of, HYTa, HYTb and HYTc. In still other embodiments, the composition comprises HYTd and two or more of, HYTa, HYTb and HYTc. The composition can also comprise HYTd, HYTa, HYTb and HYTc. In the processes
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    4/31 disclosed soil, seed, seedlings or plant foliage are in contact with HYTd or any of the above compositions.
    HYTa [029] As used in this document, the term HYTa refers to a consortium of microbes derived from samples of fertile soil and commercial sources. HYTa was deposited with American Tissue Type Culture (ATTC), Rockville, Maryland, on May 19, 2010 with an assigned deposit designation of PTA-10973.
    [030] Table 1 identifies some of the microbes in HYTa that are considered to be responsible for the beneficial effects seen when it is used to treat soil and / or foliage.
    Table 1
    Bacteria ________________________________________________________________________________________
    Bacterium
    I. Azotobacter
    1. Azotobacter vinlandii
    II. Clostridium
    1. Clostridium pasteurianum
    2. Clostridium beijerinckii
    3. Clostridium sphenoides
    4. Clostridium bifermentans
    III. Lactobacillus for my ss . paracasei 1 . Lactobacillus 2. Lactobacillus acidophillus3. Lactobacillus delbrueckii ss. Bulgaricus 4. Lactobacillus brevis
    IV. Bacillus
    1. Bacillus amyloliquefaciens (Bacillus subtilis ((SILoSil® BS))
    2. Bacillus thuringiensis var. kurstakii (Bacillus thuringiensis l Strains HD-1))
    3. Bacillus thuringiensis var. canadensis (Bacillus cereus Group)
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    5/31
    4. Bacillus pasteurii (Bacillus cereus Group)
    5. Bacillus sphaericus (Subgroup I, III, and IV)
    6. Bacillus megaterium (SubgroupA)
    V. Acetobacter or Gluconacetobacter
    1. Acetobacter aceti ss. callfaciens
    2. Acetobacter aceti ss. xylimum
    SAW. Enterococcus
    1. Enterococcus faecium (SubgroupA)
    VII. Pediococcus
    1. Pediococcus pentosaceus
    VII. Rhizobium
    1. Rhizobium japonicum
    Fungi
    I. Saccharomyces
    1. Saccharomyces cerevisiae
    II. Penicillium
    1. Penicillium roqueforti
    III. Monascus
    1. Monascus ruber
    IV. Aspergillus
    1. Aspergillus oryzae
    V. Trichoderma
    1. Trichoderma harzianum (TRICHOSIL)
    Plants
    I. Arthrospiro
    1. Arthrospira platensis
    II. Ascophyllum
    1. Ascophyllum nodosum [031] Other microorganisms contained in HYTa: Nitrobacter, Nitrosomonads, Nitrococcus, Pseudomonas, Micrococcus luteus, Actinomycete, Azotobacter vinelandii, Lactobacillus casei, Trichoderma harzianum, Bacillusomycensis lichenomormis and lichenomensis.
    [032] The microbes active in HYTa include nitrogen-fixing microorganisms native to the soil. These are Azotobacter vinelandii and Clostridium
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    6/31 pasteurianum. Bacillus-subtilis provides enzymes to break down plant waste. Bacillus cereus provides additional enzymes to break down plant residues and penicillinase for the death of unwanted bacteria. Bacillus megaterium degrades complex sugars after breaking crop residue. Lactobacillus provides food for microbes in HYTa and controls the pH of the environment. Nitrobacter organisms oxidize ammonia to nitrite (N02), while Nitrosomonas microbes oxidize nitrite to nitrate (NO3).
    [033] An important property of HYTa is the fixation of atmospheric nitrogen. The nitrogen-fixing capacity of microbes in HYTa is enhanced by the assistance of other organisms in HYTa. Nitrogen fixation requires that phosphorus (P), potassium (K) and carbon (C) are available. HYTa contains microbes that are able to decompose P, K, and C within the soil. In addition, nitrogen-fixing bacteria provide a source of nitrogen for the other microbes in HYTa.
    [034] Nitrogen fixation can occur non-symbiotically by bacteria Nitrosomonas, Nitrobacter, Azotobacter vinelandii, and Clostridium pasteurinum present in HYTa or symbiotically, as occurs in root nodules by means of Rhyzobium bacteria.
    [035] The carbon required by nitrogen-fixing microbes in HYTa is supplied by C decomposers that convert complex organic compounds in the soil into simple compounds, such as sugars, alcohols, and organic acids. C decomposers include many of the microbes identified above.
    [036] Phosphorus is necessary for nitrogen-fixing microbes to proliferate and is obtained from the metabolic activity of P decomposers that convert phosphorus immobilized in soil into a bioavailable phosphorus nutrient. P decompositors in HYTa include Azotobacter, [037] Bacillus-subtilis, Pseudomonas fluorescens and Micrococcus luteus.
    [038] The potassium required by nitrogen fixers is supplied by the K-decomposing microbes present in HYTa that activate soil potassium. The decomposers of K in HYTa include Pseudomonas fluorescens.
    [039] Three important microbes in HYTa are Bacillus subtilis (SILoSil® BS), strains of HD-1 and HD-73 Bacillus thuringiensis (SILoSil® BT), and Trichoderma harzianum (TRICHOSIL). These organisms are present in the ATTC deposit PTA-10973. They were originally obtained from Biotecnologia Agroindustrial S.A. DE C.V., Morelia, Michoacan, Mexico.
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    7/31 [040] Bacillus subtilis (SILoSil® BS) is a Gram positive bacterium that is mesophilic and grows at an optimum temperature between 25 and 35 ° C. It is aerobic and can grow in anaerobic conditions and uses a wide variety of carbon sources. It contains two nitrate reductases, one of which is used for the assimilation of nitrogen. It is capable of secreting amylase, proteases, pullulanases, chitinases, xylanases and lipases.
    [041] Bacillus thuringiensis (Strains HD-1 and HD-2 (SILoSil® BT)) are facultative anaerobic Gram positive bacteria, in the form of peritrichic flagella. The HD-1 and HD-73 strains synthesize crystals with various geometric forms of insecticidal and protein activity during the spore period. The strains HD-1 and HD-2 secrete exoquitanases when in a medium containing chitin and can be used for the degradation of crustacean residues during the production of chito-oligosaccharides.
    [042] Trichoderma harzianum (TRICHOSIL) is a saprophytic fungus. It exhibits antibiotic action and biological competition and for this reason it has biological control properties. It produces enzymes that degrade cell walls or a combination of such activities. It produces glucanases, chitinases, lipases, and extracellular proteases when it interacts with some pathogenic fungi, such as Fusarium.
    [043] As shown above, the metabolism of each group of bacteria are closely interdependent and live in close symbiotic association for proper HYTa performance.
    [044] In addition to carbon, hydrogen, phosphorus, potassium, sulfur and various trace elements, a mixture of special growth factors such as B complex, free L-amino acids and ultra-soluble trace elements is important for optimal bacterial growth. Fermentation yeasts are incorporated into HYTa to provide these components. Fixation process 2 requires large amounts of ATP. The amount of ATP naturally present is not sufficient for the fixation of biofuel 2. Yeast fermentation in HYTa compensates for the large energy deficit. During fermentation, organic acids are formed in the respiratory process and together with the phosphorus released by the P decomposers, they form the ATP. ATP is used in the biological nitrogen fixation process.
    [045] HYTa contains beneficial enzymes and soil microorganisms that replace those that have been depleted due to excessive use of chemicals that result in decreased harvests. By increasing
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    8/31 microbial activity in the soil with HYTa, the bacteria causes nutrients and microelements to be absorbed (mineralized) more efficiently and effectively by plants.
    [046] Humus is transformed by some of the microorganisms into HYTa that impregnates the soil and the plant's radical apparatus. This process provides increased nutrition for the plant. This increases the nutrients and essential elements available in the soil that can be absorbed by plants.
    [047] The use of HYTa alone or in combination with chitin, chitosan, glucosamine and / or amino acids (1) provides nutrients and elements in the soil that increase crops by 25 to 55%, (2) reduces greenhouse gas emissions greenhouse, (3) increases the efficiency of mineral fertilizers (3) reduces the use of conventional fungicides and other pesticides, (4) increases the production of plant growth regulators, (5) improves soil structure, land cultivation, and water penetration and retention, (6) cleans chemical residues and (7) shifts the pH of the soil to neutral pH.
    Microbial compositions [048] HYTa can be used, alone or in combination, with one or more components selected from the group of one or more amino acids, chitin, chitosan or glucosamine. In some cases, Acetyl-D-glucosamine may be included in the microbial composition. The microbial composition includes any and all combinations of the components mentioned above. Particularly preferred combinations include: (1) HYTa and chitin;
    [049] (2) HYTa and chitosan; (3) HYTa and glucosamine; (4) HYTa and amino acids; (5) HYTa, chitin and amino acids; (6) HYTa, chitin, chitosan and amino acids; (7) HYTa, chitosan, glucosamine and amino acids; (8) HYTa, chitosan and glucosamine and HYTa (9), chitin, chitosan, glucosamine and amino acids, the latter being particularly preferred. HYTb and in particular HYTd are the preferred sources for chitosan, glucosamine and amino acids.
    [050] When HYTa is grown in the presence of chitin, chitosan and / or amino acids, it may contain residual chitin, chitosan and / or amino acids. Under such circumstances, the HYTa culture constitutes the disclosed microbial composition and can be applied directly to the plant's soil, seed, seedlings or foliage. Alternatively, one or more of the second components can be added to complete the second components in the composition or to change its composition.
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    9/31 [051] As used in this document, the term amino acids refers to a composition containing two or more amino acids. Amino acids include tryptophan, histidine, threonine, tyrosine, valine, methionine, isoleucine, leucine, phenylalanine, lysine, aspartic acid, cysteine, glutamic acid, glutamine, serine, glycine, alanine, proline, asparagine and arginine. In preferred embodiments, amino acids are provided by using HYTb (see below).
    [052] As used in this document, the term chitin refers to a biopolymer consisting predominantly of repeating units of beta-1-4-linked Nacetyl-D-glucosamine. Chitin is found in the natural environment as a primary structural material of the exoskeleton of animals such as Arthropoda, for example, crustaceans, insects, spiders, etc., Mollusca, for example, snails, squids, etc., Coelentara, for example, organisms such as hydroids and jellyfish, Nematoda, such as non-segmented worms. Chitin is also found in several fungi, including members of the Fusarium genus. Chitin can be extracted from these natural sources by treatment with alkali, or by a process of biodegradation. The molecular weight of chitin varies according to its origin and the method of isolation. In preferred embodiments, chitin is derived as a chitin biodegradation solid containing Arthropods as described in Bioderpac applications. It is preferable that chitin has a diameter of about 50 to 75 microns to facilitate its application through drip and spray irrigation systems.
    [053] As used in this document, the term chitosan refers to a polysaccharide consisting predominantly of Dglucosamine repeat units. Chitosan is obtained by deacetylation of chitin. The degree of deacetylation compared to chitin is preferably greater than 50%, 60%, 70%, 80%, 85%, 90% and 95%. It is preferable that the level of deacetylation is sufficient to process the water-soluble chitosan at acidic pH. The molecular weight of chitosan varies depending on its origin and the method of isolation. Chitosan includes chitosan oligomers. In preferred embodiments, chitosan is precipitated at pH 9.0 of the aqueous fraction obtained from the biodegradation of chitin containing Arthropods as described in the Bioderpac applications.
    [054] As used in this document, the term chitosan oligomer refers to chitosan having 2 or more D-glucosamine repeat units and, in the case of incomplete chitin deacetylation, one or more units of N
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    10/31 acetyl-D-glucosamine. In preferred embodiments, chitosan oligomers are derived from the aqueous fraction generated in the biodegradation of chitin containing Arthropods as described in the applications of Bioderpac. In some embodiments, chitosan oligomers are used as the second component of the microbial composition.
    [055] As used in this document, the term glucosamine refers to an amino monosaccharide. In the preferred embodiments, it is the sugar residue that forms the main structure of chitin and chitosan of biopolymers. Glucosamine is present in the aqueous fraction generated during the biodegradation of chitin containing Arthropods as described in the applications of Bioderpac. Glucosamine induces plants to produce chitinase as a defense against pathogens containing chitin.
    HYTb and HYTc [056] As used in this document, the term HYTb refers to the aqueous fraction and HYTc refers to the solid fraction obtained from the biodegradation of chitin containing Arthropods as described in US Patent Application Serial No. 61 / 289,706, filed 12/23/09, entitled Biodegradation of Crustacean Byproducts, US Patent Application Serial No. 61 / 299,869, filed on 1/29/10 entitled Biodegradation Process and Microbial Composition and US Patent Application Serial 61 / 355,365 filed on 06/16/2010 entitled Biodegradation Process and Composition and PCT / EP2010 / 070285 filed on 12/20/2010 entitled “Biodegradation Process and Composition”, each of which is incorporated by reference in this document in its entirety.
    [057] Briefly, in the process of arthropod biodegradation a microbial composition is used to degrade the arthropod or residual components of the arthropod. It is a lactic acid fermentation process. The microbial composition contains microbes that produce enzymes that can degrade chitin containing components from arthropod to chitin, chitosan, N-acetyl glucosamine and glucosamine. It also contains microbes that produce enzymes that can break down proteins and fats to produce amino acids and lipids.
    [058] A preferred microbial composition for arthropod degradation is referred to as HQE. HQE was deposited with the American Type Culture Collection (ATCC) Manassas, VA, USA on April 27, 2010 and given the Patent Filing Designation PTA-10861.
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    11/31 [059] In a preferred embodiment, the marine arthropod is a crustacean and the preferred crustacean is shrimp. The shrimp by-product comprises cephalothorax and / or shrimp exoskeleton.
    [060] In the biodegradation process, it is preferable that the fermentation is an optional aerobic fermentation. It is also preferable that the fermentation is carried out at a temperature of about 30 ° C to 40 ° C. The pH is preferably less than about 6, more preferably less than about 5.5. However, the pH should be kept above about 4.3. Fermentation is carried out for about 24 to 96 hours. In some embodiments, fermentation is carried out for about 24 to 48 hours and more preferably 24 to 36 hours. These fermentation periods are much shorter than the typical fermentation periods of the prior art from 10 to 15 days to achieve substantially the same amount of digestion, although without detectable chitosan and glucosamine formation.
    [061] The mixture is preferably separated by centrifugation. (for example, about 920 g). Gravity separation can also be used, but it is not preferred because of the time required to achieve separation.
    [062] The mixture separates into three fractions: solid, aqueous and lipid. The solid fraction comprises chitin and is called HYTc. The aqueous fraction comprises protein hydrolyzate, amino acids, chitosan and glucosamine and is called HYTb. The lipid fraction comprises sterols, vitamins A and E and carotenoid pigments such as astaxanthin.
    [063] It is preferable that HQE is used in the biodegradation process. In other embodiments, it is preferable that HYTb previously prepared is added to HQE or to the fermentation broth. As described above, HYTb contains amino acids, chitosan, glucosamine and trace elements including calcium, magnesium, zinc, copper, iron and manganese. HYTb also contains enzymes such as lactic enzymes, proteases, lipases, chitinases, lactic acid, polypeptides and other carbohydrates. HYTb can also contain latent microorganisms from a previous biodegradation process. These microorganisms can be reactivated and, in combination with HQE, contribute to a more resistant biodegradation process compared to when HQE is used alone as otherwise described in this document.
    [064] More particularly, the process includes the following steps:
    The. Activation of microbial cells in a sugar-based solution to improve their growth and biomass formation. B. Grinding of
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    12/31 shrimp by-products (cephalothorax and exoskeleton) to produce a homogeneous paste.
    ç. Homogeneous mixture of the shrimp by-product slurry in at least 10% of the activated inoculum.
    d. Adjust pH values to less than 6.0 in the mixture using a citric acid solution to inhibit the growth of microorganisms and promote the development of microbial cells that make up the inoculum.
    and. Fermentation of the mixture in a non-continuous stirred system at temperatures within a range of 30 to 40 ° C for at least 96 hours, maintaining the pH at less than
    5.0. The pH is monitored periodically. If the pH is raised above 5.0, a citric acid buffer is added in an amount to maintain the pH below 5.0.
    f. Centrifuging the yeast to separate the three main fractions: chitin, liquid hydrolyzate and pigmented paste.
    g. Washing the raw chitin and collecting the rinse water to recover fine solids or minerals.
    H. Chitin drying and storage.
    i. Drying and storage of the liquid hydrolyzate.
    j. The pigmented paste (lipid fraction) is stored in closed containers for conservation.
    [065] The process and operational fundamentals are best understood with reference to the following detailed description.
    Activation of microbial cells [066] A microbial composition as disclosed in this document is used as an inoculum. The HQE inoculum has a microbial concentration of about 2.5 to 3.0% (w / v). HQE is activated by dilution to 5% in sugar cane solution (3.75% of final concentration of sugar cane), and incubated at 37 ° C for 5 days. HYTb (10 ml per liter of culture) is preferably added to provide a source of minerals and is naturally derived from minerals. The cell growth of microorganisms was estimated by the optical density measured at 540 nm. Activation is completed at an optical density of about 1.7. The concentration of microbes after activation is about 1.9 to 3.0% (w / v).
    Sample preparation [067] Samples of shrimp by-products are obtained from shrimp processing plants. Slightly defrosted and chopped residue (1500
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    13/31 g per batch) is mixed with 99 grams of sugarcane (final concentration of 6.6% by weight) and 85.5 ml of activated HQE 5% (v / w) (optical cell density = 1, 7). The pH is adjusted to 5.5 using 2 M citric acid.
    Fermentation control [068] The mixture is incubated at 36 ° C, with non-continuous shaking for 96 h. During the fermentation process, the pH is monitored by means of a potentiometer, and the total titratable acidity (ATT,%) was determined by titration with 0.1 N NaOH until a pH of 8.5 is obtained. ATT is expressed as a percentage of lactic acid.
    Separation conditions [069] The fermentation product is a viscous silage that has an intense orange color, due to the presence of astaxanthin. Silage is centrifuged (5 ° C) at 1250 rpm (930g) for 15 min to obtain chitin, liquid hydrolysates, and pigment paste. The upper phase (pigment paste) is separated manually. The liquid hydrolysates are separated by decantation, and the sediment that constitutes the crude chitin is washed with distilled water to separate the fine solids. The resulting liquid is collected and dried. The crude chitin, liquid hydrolysates and fine solids are dried at 60 ° C. All fractions are stored to protect them from light.
    [070] Other microbial compositions for the production of HYTb and HYTc are set out in the following table 2.
    Table 2
    Culture Composition
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    14/31
    Microorganism 1 2 3 4 5 6 7 8 9 10 Bacillus subtilis X X X XX X XX Bacillus cereus X XXXXX Bacillus megaterium X X Azotobactervinelandii X XXXXX Lactobacillusacidophilus X X X X XX X XLactobacillusgot married X XX X X XTrichodermaharzianum X X X XX X XX Rhizobium X XXXXX
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    15/31
    Microorganism 1 2 3 4 5 6 7 8 9 10 japonicum Clostridium pasteurianum X X X X X X Bacilluslicheniformis X X XX X XX X Pseudomonasfluorescens X X XX X Bacillusthuringiensis X X X X X X Streptomyces X X X X X X X Nitrobacter XX X X X Micrococcus XX X X X Proteus vulgaris XX X X X
    [071] These microorganisms are preferably derived from HQE and are referred to as Bacillus subtilis ((SILoSil® BS), Bacillus cereus (Bioderpac, 2008), Bacillus megaterium (Bioderpac, 2008), Azotobacter vinelandii (Bioderpac, 2008), Lactobacillus acidophilus (Bioderpac, 2008), Lactobacillus casei (Bioderpac, 2008), Trichoderma harzianum (TRICHOSIL), Rhizobium japonicum (Bioderpac, 2008), Clostridium pasteurianum (Bioderpac, 2008), Bacillus licheniformis (Bioderpac, 2008), HD-1 and HD-73 strains of Bacillus thuringiensis (SILoSil® BT), Streptomyces (Bioderpac, 2008), Micrococcus (Bioderpac, 2008), Nitrobacter (Bioderpac, 2008) and Proteus (Bioderpac, 2008). such organisms can be easily isolated from HQE and recombined to form the disclosed microbial composition to degrade arthropods to produce HYTb and HYTc.
    HYTb [072] HYTb contains amino acids (about 12% by weight), chitosan (about 1.2% by weight), glucosamine (about 1% by weight) and trace elements (about
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    16/31 of 6% by weight), including calcium, magnesium, zinc, copper, iron and manganese. It also contains enzymes such as lactic enzymes, proteases, lipases, chitinases, among others, lactic acid, polypeptides and other carbohydrates. The specific gravity of HYTb is usually about 1.050 to 1.054. The average amino acid content in HYTb for certain amino acids is set out in Table 2.
    Table 3
    Amino Acid Powder hydrolysatesdry Aspartic acidGlutamic acid Serine Histidine Glycine Threonine Alanine Proline Tyrosine ArginineValine Methionine Isoleucine Tryptophan Leucine 3839169281436.125.87022.22016.418.33.123 Amino Acid Powder hydrolysatesdry Lysine phenylalanine 3913 Total 431
    [073] In some embodiments, HYTb may constitute a second component that is combined with HYTa or used separately as a soil concealer and / or as a foliage spray.
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    17/31
    HYTc [074] The primary component of HYTc is chitin. It has an average molecular weight of about 2300 daltons and constitutes about 64% by weight of the composition. About 6% of HYTc contains minerals, including calcium, magnesium, zinc, copper, iron and manganese, about 24% by weight of protein and 6% of water. It has a specific gravity of about 272 kg / m3. In some embodiments, HYTc may constitute a second component that is combined with HYTa or used separately as a soil concealer and / or as a foliage spray.
    [075] HYTa is preferably used with HYTb and HYTc either in combination, or separately as a soil concealer or foliage spray.
    [076] The microbes in HYTa require the trace elements calcium, magnesium, sulfur, boron, manganese, zinc, molybdenum, iron, copper, sodium and silicon. These important trace elements can often be obtained from toxic chemical reactions, which are not suitable for certified organic products. Accordingly, it is preferable that these trace elements are obtained from an organic source such as HYTb and / or HYTc.
    HYTd [077] HYTd is obtained by fermenting chitin with a microbial composition such as HQE suspended in HYTb. The process is similar to that described above for the production of HYTb and HYTc, except that the substrate is chitin, for example, HYTc, instead of chitin containing Arthropods.
    [078] Figure 12 is a flow chart, showing the digestion of the crustacean to form HYTb and HYTc. HYTc and HYTb are subsequently processed with HQE to form HYTd, a solution with relatively high amounts of chitosan and glucosamine compared to HYTb.
    [079] Figure 13 is a flowchart showing the digestion of fungi, including filamentous fungi, yeasts and / or insects to form HYTb and HYTc. HYTc and HYTb are further processed with HQE to form HYTd.
    [080] HYTb already contains chitosan (about 0.5 to 1.5% by weight) and glucosamine (about 0.5 to 1.5% by weight). The amount of chitosan and glucosamine in HYTd ranges from about 2% by weight to 2.5% by weight of chitosan and from about 2% by weight to 5% by weight of glucosamine. This represents an increase in the amount of chitosan and glucosamine in
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    18/31 compared with HYTb of about 0.5% by weight to 2.5% by weight of chitosan and from about 0.5% by weight to 5% by weight of glucosamine.
    [081] HYTd when undiluted is similar to HYTb, but contains higher amounts of chitosan and glucosamine. HYTd contains amino acids (about 5 to 12% by weight) and trace elements (about 6% by weight), including calcium, magnesium, zinc, copper, iron and manganese. It also contains enzymes such as lactic enzymes, proteases, lipases, chitinases, among others, lactic acid, polypeptides and other carbohydrates. In some embodiments, the degree of acetylation of the chitosan produced is 20% or less, preferably 15% or less, more preferably 10% or less, even more preferably 8% or less and more preferably 5% or less. The average amino acid content in HYTd for certain amino acids is similar to that of HYTb. See Table 3.
    [082] Preferred HYTd comprises 12% by weight of L-amino acids [083] (Aspartic acid, Glutamic acid, Serine, Histidine, Glycine, Threonine, Alanine, Proline, Arginine, Valine, Methionine, Isoleucine, Tryptophan, Phenylalanine, Lysine and Threonine) and 5% by weight of glucosamine and chitosan. HYTd also preferable contains one or more or all of the soluble minerals (P, Ca, Mg, Zn, Fe and Cu), enzymes and lactic acid from the chitin digestion process, as well as other polysaccharides.
    [084] As used in this document, the term glucosamine includes glucosamine or a mixture of glucosamine and N-acetyl glucosamine. In most embodiments, HYTd contains glucosamine and N-acetyl glucosamine.
    [085] HYTd may also contain particulate chitin that has not been fully digested. In general, the fermentation mixture is filtered to remove large chitin particles. The filtrate generally contains no more than 2% by weight of chitin.
    Activation of HYTa [086] The aforementioned microbial compositions can be used to treat soil, seeds, seedlings and / or plant foliage.
    [087] However, HYTa is first activated before use.
    [088] In preferred embodiments, HYTa is activated by incubating an inoculum of HYTa in aqueous solution for 24 to 168 hours to allow microbes to grow and reproduce before being used in the process of treating soil, seeds, seedlings and / or plant foliage. Incubation conditions influence the initial global properties of HYTa.
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    19/31 [089] In one embodiment, an HYTa inoculum is diluted in water at a ratio of 1/100 and allowed to incubate at a temperature of approximately 36 ° C at a pH of 6.8 to 7.1 for approximately 24 to about 168 hours (7 days). HYTb can optionally be used during this activation. The nitrogen fixing microbes Azotobacter vinelandii and Clostridium pasteurianum proliferate under reduced nitrogen growth conditions. In addition, as the oxygen concentration decreases, Lactobacilli, including Lactobacillus acidophilus and Lactobacillus casei, proliferate. Colony forming units (CFUs) for some of the bacteria in activated HYTa are set out in table 3:
    Table 4
    Azotobactervinelandii 101,050,000Cfu / mL Clostridium 104,275,000 pasteurianum Cfu / mL Bacillus subtilis 1,100,000Cfu / mL Bacillus cereus 25,000 Cfu / mL Bacillus megaterium 10,000 Cfu / mL Lactobacillus 500,000 Cfu / mL Nitrobacter 5,000 Cfu / mL Nitrosomonas 2,500 Cfu / mL Total 206,967,000
    Cfu / mL [090] The HYTa obtained after this incubation maintains the beneficial properties of HYTa, but is particularly adapted as a soil corrective for the treatment of soils lacking in nitrogen given the nitrogen fixing capabilities of Azotobacter vinelandii and Clostridium pasteurianum .
    [091] If soil pathogens such as filamentous fungi of the genus Fusarium or nematodes are present, or considered to be present, HYTa can be activated in substantially the same conditions, but in the presence of chitin. Chitin stimulates the expansion of microbes
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    20/31 responsive to chitin such as Pseudomonas fluorescens, Trichoderma harzianum, Bacillus thuringiensis, Streptomyces sp. Nitrobacter sp. Micrococcus sp. and Bacillus subtilis. HYTa obtained in these conditions has antifungal, fungicidal, antinematoid, nematodicidal and insecticidal properties insofar as such pathogens contain chitin. Such microbial compositions can be applied directly to soil or seed, seedlings and / or plant foliage. Such microbial compositions also have the ability to fix nitrogen as in the incubation mentioned above in the absence of chitin.
    [092] In addition to chitin incubation, HYTa can be activated with chitin and amino acids. A preferred source of chitin is HYTc. When HYTc is used the protein and minerals in HYTc are also present during activation.
    [093] In addition, HYTa can be activated in the presence of amino acids and chitosan. A preferred source of amino acids and chitosan is HYTb and / or HYTd. When HYTb and / or HYTd is used glucosamine and other components of HYTb and / or HYTd are also present during activation.
    [094] Optionally, HYTa can be incubated with chitin, amino acids and chitosan. A preferred source of chitin is HYTc. A preferred source for amino acids and chitosan is HYTb and / or HYTd. When HYTb, HYTd and HYTc are used, other components in these formulations are also present during activation.
    Use of Activated HYTa [095] Activated HYTa can be used alone or in combination with other components, such as chitin, chitosan (for example, HYTc), glucosamine and amino acids (for example, HYTb or HYTd) to treat soil, seed, seedlings or foliage. In some embodiments, combinations of these components can be applied as a mixture. In other modalities, they can be applied separately. In yet other modalities, the components can be applied at different times.
    [096] In one embodiment, activated HYTa can be applied to soil, seeds or seedlings, or used in foliar applications by direct application to foliage. However, when plant pathogens are present, it is preferable that the microbial composition comprises activated HYTa, chitin and / or chitosan. Alternatively, HYTa can be activated in the presence of chitin. Chitosan is known to have bactericidal, fungicidal and antiviral properties, as well as its ability to stimulate plant growth and induce
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    21/31 plant resistance to pathogens. In other embodiments, glucosamine is a part of the microbial composition.
    [097] In a preferred embodiment, HYTa activated alone or in combination with chitin (preferably HYTc), and / or chitin, chitosan and amino acids (preferably HYTb, HYTd and / or HYTc) is applied to soil, seeds, seedlings and / or foliage. It is preferable that HYTa is used in combination with chitin, chitosan, glucosamine and amino acids. HYTc is the preferred source of chitin, while HYTb and / or HYTd are the preferred sources of chitosan, glucosamine and amino acids, however, the components of the microbial composition particularly HYTa, chitin, chitosan, glucosamine and amino acids can be applied separately or in any combination or subcombination. They can be applied simultaneously or sequentially, in any given order. However, the preferred mode of application is initially to apply everything at the same time. The application of the preceding components provides the direct treatment of plant pathogens, the induction of pathways of resistance to the plant pathogen, and the nutrition of HYTa microbes, the indigenous non-pathogenic soil flora, and the plant.
    [098] When the soil is initially treated with a microbial composition comprising HYTa activated alone, the microbes present in the composition have an opportunity to fill the soil and change its taxonomic composition. In some situations, initial HYTa colonization provides little or no nutrients for the plant. In such cases, it is important to maintain a reserve of nutrients to support the growth of microbes by colonizing the rhizosphere and the growth of plants in the soil. It may be necessary to repeat the application of HYTa, depending on the growth cycle and the nutritional regime of the plant. In other cases, it may be sufficient to provide additional applications of amino acids, chitin and / or chitosan, for example, HYTb and HYTc, to the previously treated soil.
    [099] When HYTa is used in combination with, for example, HYTb, HYTd and / or HYTc, additional nutrients are available for HYTa microbes and plants present in the treated soil.
    [100] Table 5 sets out a typical fourteen-week program for the application of HYTa, HYTb and HYTc for drip-irrigated crops grown in the soil. Values are per hectare. For HYTa and HYTb, the values represent liters per week. For HYTc, the values represent kilograms per week.
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    Table 5
    WeekHYT-A M® .. y ',3 1 '1 ··· ι ,, · ϋ · , |, · Ί · ........ peW8ÍW10 ......... ’ι'ι'ρ., Ιιμι '1 W13 W14 0 0 1 0 1 0 1 0 1 0 1 0 1 HYT-B 10 5 0 3 2 3 2 3 2 3 2 3 2 3 HYT-C 1 1 1 1
    [101] The pulse in which the microbial composition is injected into the irrigation system must be one in which the microbial composition is able to reach the root system and remain there overnight, while the system is off. For maximum HYTc performance, it should be applied at the same time as a mixture with HYTa. The protocol must be continued, while the plant remains in production. This protocol covers all stages of the plant, including germination, root formation, plant growth, flowering, fruiting, fruit formation harvest and harvest. This protocol is designed for the maximum potential yield covering nutritional aspects, aspects of biostimulation and protection against diseases such as nematodes and fungi.
    [102] The process can be carried out by placing it in contact with the soil to form a treated soil. In some cases, the process is repeated. In some cases, plants, seedlings or seeds are already present in the soil before treatment with the microbial composition. In other cases, plants, seedlings or seeds are transplanted into the soil after treatment with the microbial composition.
    [103] In general, before application the number of hectares or acres to be treated is determined. Then, the recommended amount of activated HYTa per hectare or acre is multiplied by the area to be treated and diluted in enough water to irrigate or spray the soil or crops in the area to be treated. The same procedure can be followed for HYTb and / or liquid HYTd. HYTc, being a solid, can be applied directly as a solid or as a suspension in water. HYTc is preferably ground into micron size particles before use.
    [104] The process can be carried out with infertile soil. Such soils are generally those that had at least one with low cation exchange capacity, low water holding capacity, low matter content
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    23/31 organic, and low levels of available nutrients are present. In general, infertile soil does not support vigorous plant growth and / or produces low yields.
    [105] For non-soil systems such as hydroponics, the same protocol applies, but with a daily distribution following the fertigation program.
    [106] Microbial compositions can be used in connection with any plant, including, but not limited to, alfalfa, banana, barley, broccoli, carrots, corn, cucumber, garlic, grapes, leek, melon, onion, potato, raspberry , rice, soy, pumpkin, strawberry, sugar cane, tomato and watermelon.
    [107] When applied as a soil concealer, the microbial composition containing HYTa, chitin, amino acids and chitosan increases crop production by an average of about 25% to 55% compared to the observed 15 to 25% increase in crop production for E2001. From Karl Co. SA de CV, Navojoa, Sonora, Mexico.
    [108] Microbial composites can also result in a decrease in the amount of chitin used. For example, chitin has been used as a soil concealer in the prior art. Typically, about 600 kg of chitin was used per hectare. However, the beneficial effects of such use have not been seen for up to six months. When HYTa was activated in the presence of chitin and then combined with chitin and applied as a soil corrective, the beneficial effects were observed after seven days, using only 4 to 6 kg of chitin per hectare.
    [109] Although the disclosure is primarily directed towards the use of the disclosed microbial compositions, HYTb, HYTc and / or HYTd for agricultural applications, such compositions or their components and processes can also be used in horticultural applications to improve flower production and foliage and decrease the use of conventional fungicides and insecticides.
    [110] When activated HYTd and HYTa, HYTb, or HYTc is applied to soil, seedlings or foliage, this form treated soil, treated seed, treated seedling, treated foliage and treated plants. HYTd is also a new composition. Therefore, the soil, seed, seedlings, foliage and plants treated with HYTd and HYTa, HYTb and / or HYTc are also new. Since HYTd, HYTa, HYTb and HYTc are usually diluted before application, the soil, seed, seedlings and foliage will normally contain the components of HYTd, HYTa, HYTb and / or HYTc in diluted form.
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    24/31 [111] Soil treated with HYTa is defined as soil containing one or more microbes that are unique to HYTa dispersed in treated soil. Such microbes can be detected in genetically treated soil using a BioChip that detects microbial populations based on DNA. See, for example, US Patent Publication 2007/0015175, incorporated herein by reference. Other methods, such as PCR, which are known to those skilled in the art can also be used. Microbes in HYTa that are particularly preferred are Bacillus subtilis (SILoSil ® BS), HD1 strain of Bacillus thuringiensis, strain HD-73 of Bacillus thuringiensis (SILoSil® BT) and Trichoderma harzianum (TRICHOSIL) each of which can be isolated from deposit of HYTa or obtained from Biotechnology Agroindustrial SA DE CV, Morelia, Michoacan, Mexico. Trichoderma harzianum (TRICHOSIL) is preferable, since it is important during the activation of HYTa in that it causes intercomponent synergies between the other microbes in HYTa. The identification of one or more of these microorganisms can be further combined with the identification of other microbes in HYTa, if necessary, to confirm the presence of HYTa or that HYTa was present. Trichoderma harzianum (TRICHOSIL) was filed with ATCC on October 6, 2011 and given Patent Deposit Designation PTA-12152. Bacillus subtilis (SILoSil ® BS) was filed with ATCC on October 7, 2011 and given Patent Deposit Designation PTA-12153. The HD-1 and HD-73 strains of Bacillus thuringiensis (SILoSil ® BT) were deposited with ATCC on May 31, 2012 and given Patent Deposit Designation PTA-12967.
    [112] The seed, seedlings, foliage and treated plants are defined in a similar way. In these cases, HYTa microbes are found on the surface of the seed, seedlings, foliage and treated plants.
    [113] As used in this document, the term essentially consisting of in connection with HYTa, HYTb and HYTc means any HYTa, HYTb and / or HYTc alone or in combination without additional microbes.
    Process for producing HYTd [114] The process for producing HYTd is disclosed in US Patent Application Serial No. 61 / 500,527 filed on June 23, 2011 entitled “Process for Making Chitin and Chitin Derivatives” which is expressly incorporated into this document by reference.
    [115] Soon, HQE or related chitin degradation microbial compositions are activated and added to HYTb. Solid chitin is
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    25/31 is added and the mixture is fermented for 3 to 7 days. Chitin can be obtained from HYTc or other sources such as chemical treatment or biodegradation of chitin containing fungi, filamentous fungi, yeasts and / or insects. HYTc is the preferred source of chitin. Chitin is preferably micronized. Micronized chitin or residual chitin can be used.
    Use of HYTd [116] HYTd can be used as a biostimulant I for root and foliage growth and as a fungicide.
    [117] If used alone as a fungicide, it is preferable that 20 liters are applied per hectare.
    [118] If used to treat stressed plants, it is preferable for HYTd to be applied at 3 to 10 liters per hectare.
    [119] HYTd can also be applied at 3 to 5 liters per hectare.
    [120] HYTd can be applied directly to soil, foliage or both.
    [121] HYTd can be used in conjunction with other components, such as HYTb, HYTc and / or HYTa. When used with other components, HYTd can be combined with the component to form new compositions. Such compositions can be applied to the estate or plant. Alternatively, HYTd and one or more of HYTa, HYTb and / or HYTc can be applied separately or at different times.
    Example 1 [122] The following protocol was applied to the soil of asparagus plants.
    Table 6
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    test HYTa HYTb HYTç HYTd 1 0 0 0 0 2 3 liters / hectare and then 1 liter / hectare every 45 days. 0 0 0 3 3 liters / hectare and then 1 liter / hectare every 45 days. 2 liters / hectare and then 1 liter / hectare every 12 days. 0 0 4 3 liters / hectare and then 1 liter / hectare every 45 days. 5 liters / hectare and then 2 liters / hectare every 12 days. 0 5 liters / hectare and then 2 liters / hectare every 12 days.
    [123] The results are shown in figures 1 and 2. As can be seen without any application of any HYT product a relatively small plant is obtained with the development of the poor root. As you progress through 2, 3 and 4 tests, it is clear that each treatment results in better leaf and root development.
    Example 2 [124] This example demonstrates the benefit of treating potatoes with HYTa compared to a control.
    [125] The soil contained 1% or less of organic matter and had a pH between 7.3 and 7.5. The following per hectare was applied throughout the test:
    400 to 600 kg of ammonium sulfate.
    -250 to 400 kg of 11-52-0, phosphorus.
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    300 kg of potassium sulfate
    150 to 200 kg of potassium nitrate to 100 kg of magnesium sulphate to 50 kg of zinc sulphate
    -HYTa-10L [126] Two liters of HYTa were applied to each hectare of soil with the first application of water. At planting, 4 liters of HYTa per hectare were applied to the soil. After the development of the stall, 2 liters of HYTa per hectare were applied to the soil. When the potatoes were about 4 cm in size, 2 liters of HYTa per hectare was applied to the soil.
    [127] The results are shown in Table 7.
    Table 7
    TESTHYT + A CONTROL size parts kilos parts kilos giant 0 0 00 0 0.00 first 4 1 20 1 0.30 second 18 3 33 11 1.93 third 21 2.74 25 337 bedroom 71 3.24 118 5.27 MONO 6 0.79 7 1.30 TOTAL 120 11.3 162 12.1
    DIFFERENCE parts kilos 0 Ü06 3 0 91 7 1 41 • 4 -0 63 47 -2.03 -1 -0.51 -42 -0.86
    % IN RELATION TO CONTROL parts kilos 300% 307% 64% 73% -16% -19% -40% -39% -14% -39% -26% -7%
    [128] The most valuable giant potatoes Potatoes are followed by 1, 2 and 3 sizes. The next two sizes can be used for potatoes or processed seeds. Figures 3 and 4 graphically present the results in table 7. As can be seen, there is a significant increase in the number and mass of potatoes in the first and second sizes. Figure 5 contains photographs comparing the potatoes obtained.
    Example 3 [129] This example demonstrates the benefit of treating potatoes with HYTa, HYTb, HYTc and HYTd compared to treatment with HYTa.
    [130] The soil contained 1% or less of organic matter and had a pH between 7.3 and 7.5. The following per hectare was applied throughout the test:
    400 to 600 kg of ammonium sulfate.
    250 to 400 kg of 11 -52-0, phosphorus.
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    300 kg of potassium sulfate
    150 to 200 kg of potassium nitrate to 100 kg of magnesium sulphate to 50 kg of zinc sulphate
    -HYTa-10L
    HYTb- 8 L
    HYTc - 4 kg
    HYTd- 5 L [131] Two liters of HYTa were applied to each hectare of soil with the first application of water. At planting, 4 liters of HYTa, 3 kilograms of HYTc and 5 liters of HYTd per hectare were applied to the soil. After the development of the stall, 2 liters of HYTa per hectare were applied to the soil. When the potatoes were about 4 cm in size, 2 liters of HYTa per hectare was applied to the soil.
    [132] After the beginning of tuber formation, 1 liter of HYTb was applied to the leaves of the plant every 6 to 10 days. This was repeated eight times.
    [133] The results are shown in Table 8.
    Table 8
    TESTHYT + A + B + C + D CONTROL size parts kilos parts kilos giant 4 I 54 0 ooo first 8 2 07 1 030 second 19 360 11 1 93 third 32 404 25 3.37 guarto 39 169 118 527 MONO 1 1 54 7 1.30 TOTAL 103 14.5 162 12.1
    DIFFERENCE parts kilos 4 15-1 7 1 78 8 168 7 0 68 -79 -358 -6 0 24 -59 2.32
    % IN RELATION TO CONTROL parts kilos 700% 602% 73% 87% 28% 20% -67% -68% -86% 19% 36% 19%
    [134] Figures 6 and 7 graphically present the results in Table 8. As can be seen, there is a significant increase in the number and mass of potatoes in the giant up to third sizes compared to treatment with HYTa alone. Figure 8 contains photographs comparing the potatoes obtained.
    Example 4 [135] This example demonstrates the benefit of treating potatoes with HYTa, HYTb, HYTc and HYTd compared to treatment with Metam Sodium in a fungal infested soil. The results are shown in table 9.
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    29/31 soil was treated as described in example 3. HYTa, HYTc and HYTd were applied as set out in example 2. Three hundred liters of Metam Sodium were applied per hectare.
    Table 9
    TESTHYT + A + B + C + D CONTROL Metam-Sodium size parts kilos parts kilos giant 4 1 1 0.43 first 8 2.07 1 0.32 second 19 3.60 35 6.46 third 32 404 24 2.57 bedroom 39 169 88 3.53 MONO 1 1.54 6 147 TOTAL 103 14.5 155 15.0
    <% compared to control parts kilos 300% 257% 700% 547% -46% -44% 33% 52% -55% -54% -83% 4% -34% -3%
    [136] Figures 9 and 10 graphically show the results in Table 9. As can be seen there is a significant increase in the number and mass of potatoes in the giant, first and third sizes compared to the treatment with Metam-Sodium. Figure 11 contains photographs comparing the potatoes obtained.
    [137] Metam sodium is a soil fumigant used as a pesticide, herbicide and fungicide. Its use is illegal, some countries due to environmental concerns. Treatment with HYTa, HYTb, HYTc and HYTd can eliminate the use of Metam Sodium in certain applications, thus reducing its environmental impact and the cost of using this soil fumigant.
    Example 5 [138] This example demonstrates the effect of treatment with cucumber HYTa and HYTc infested with the nematode Rhabditis and the fungus Fusahum oxisporum.
    [139] Ten liters of HYTa and 3 kilograms of HYTc per hectare were applied to the soil. This protocol was repeated 8 days later.
    [140] The results are shown in table 10.
    Table 10
    Before After 18 days
    - Nematode population per kg of soilper Kg. of Soil Nematode population per kg of soilNematodes per Kg. Of
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    Nematode Low<100 Average> 100 <500 High> 500 Low<100 Average> 100 <500 High> 500 Rhabditis 9,600 850Nematode population per kg of soil Nematode population per kg of soil Fungus Low400 Average> 600 High> 700 Low400 Average> 600 High> 700 Fusarium oxisporum 770 3
    [141] As can be seen, the population of these organisms has been substantially reduced 18 days after treatment.
    Example 6 [142] This example demonstrates the effect of treatment with HYTa, HYTc and HYTd on tomatoes infested with the fungus Fusarium oxisporum.
    [143] Five liters of HYTa, 1 kilogram of HYTc and 5 liters of HYTd per hectare were applied to the soil. This treatment was repeated every 15 days [144] The results are shown in table 11.
    Table 11
    Colony Forming Unit (UFC / g) HYTA, C + D Control Fusarium oxysporum 500 1,666 Rhizoctonia solani 0Phytopthora sp. 0Pythium sp 0
    [145] As can be seen, treatment with HYTa, HYTc and HYTd significantly reduced the colony forming unit of the fungus.
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    Example 6 [146] The foliage of the mold-infested tomato plant (fungus Phytophthora infestans) was treated with HYTd. Two, 4, 6 and 8 liters of HYTd were diluted in 100 liters of water per hectare. The control was untreated tomatoes. After a week, the infestation was stopped. The fungus in the control crop however developed a cotton-like appearance that resulted in damage to the plant and the development of necrosis (results not shown). The optimum amount of HYTd in the treatment of Phytophthora infestans is 6 liters per hectare.
    权利要求:
    Claims (12)
    [1]
    1. Composition, characterized by the fact that it comprises HYTd, in which said HYTd comprises the liquid fraction obtained from the fermentation of HYTb and HYTc with HQE for 3 to 7 days, in which said HQE is a composition comprising the designation of ATCC Patent Deposit PTA-10861, in which said HYTb comprises the liquid fraction obtained from the fermentation of Arthropods containing chitin with 10% activated HQE with non-continuous agitation at 36 ° C and pH below 5 for 96 hours, and in which said HYTc comprises the solid fraction obtained from the fermentation of Arthropods containing chitin with activated HQE 10% with non-continuous stirring at 36 ° C and pH below 5 for 96 hours, and in which the activated HQE is obtained by diluting a HQE inoculum with microbes from 2.5-3.0% (w / v) to 5% in sugar cane solution and incubated at 37 ° C for 5 days.
    [2]
    2. Composition according to claim 1, characterized by the fact that it comprises at least one of HYTa, HYTb and HYTc, wherein said HYTa comprises a composition comprising the ATCC Patent Filing designation PTA-10973.
    [3]
    Composition according to claim 1, characterized by the fact that it additionally comprises two or more of HYTa, HYTb and HYTc, wherein said HYTa comprises a composition comprising the ATCC Patent Filing designation PTA-10973.
    [4]
    4. Composition according to claim 1, characterized by the fact that it additionally comprises HYTa, HYTb and HYTc, wherein said HYTa comprises a composition comprising the ATCC Patent Filing designation PTA-10973.
    [5]
    5. Process, characterized by the fact that it comprises putting the soil, seed, seedlings or plant foliage in contact with the components of the composition, as defined in any one of claims 1 to 4.
    [6]
    6. Process, according to claim 5, characterized by the fact that it additionally comprises putting in contact the soil, seeds, seedlings
    Petition 870190042228, of 05/06/2019, p. 41/43
    2/2 or plant foliage with at least one of HYTa, HYTb and HYTc, wherein said HYTa comprises a composition comprising the ATCC Patent Filing designation PTA-10973.
    [7]
    7. Process, according to claim 5, characterized by the fact that it additionally comprises putting in contact the soil, seed, seedlings or plant foliage with two or more of HYTa, HYTb and HYTc, wherein said HYTa comprises a composition comprising the ATCC Patent Filing designation PTA-10973.
    [8]
    8. Process according to claim 5, characterized by the fact that it additionally comprises contacting the soil, seed, seedlings or plant foliage with HYTa, HYTb and HYTc, wherein said HYTa comprises a composition comprising the designation of ATCC Patent Filing PTA-10973.
    [9]
    9. Process, characterized by the fact that it comprises combining HYTd and at least one of HYTa, HYTb and HYTc to form a mixture, as defined in any of claims 2 to 4.
    [10]
    10. Process according to claim 9, characterized by the fact that it further comprises applying said mixture to the soil, foliage, seed or seedlings.
    [11]
    11. Treated soil, characterized by the fact that it is treated with the composition, as defined in any one of claims 1 to 4.
    [12]
    12. Process, characterized by the fact that it comprises planting seeds, seedlings or plants in treated soil, as defined in claim 11.
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    AU2016224901A1|2017-08-31|Microbial consortia
    同族专利:
    公开号 | 公开日
    RU2583302C2|2016-05-10|
    CA2840138C|2017-01-17|
    UA113736C2|2017-03-10|
    CN104735983A|2015-06-24|
    ES2791827T3|2020-11-06|
    CN104735983B|2017-04-26|
    EP2723172B1|2020-03-18|
    AU2015205815B2|2016-01-21|
    AU2015205815A1|2015-09-17|
    AU2012273927B2|2015-04-30|
    US20150119244A1|2015-04-30|
    CA2840138A1|2012-12-27|
    US20120329650A1|2012-12-27|
    BR112013033319A2|2016-08-16|
    AU2012273927A1|2014-02-13|
    RU2014102002A|2015-08-27|
    RU2016111734A|2018-11-27|
    WO2012175739A1|2012-12-27|
    US10617123B2|2020-04-14|
    US20170215431A1|2017-08-03|
    EP2723172A1|2014-04-30|
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    法律状态:
    2016-10-11| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|
    2018-07-17| B07A| Application suspended after technical examination (opinion) [chapter 7.1 patent gazette]|
    2019-02-05| B07A| Application suspended after technical examination (opinion) [chapter 7.1 patent gazette]|
    2019-08-13| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|
    2019-10-08| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 25/06/2012, OBSERVADAS AS CONDICOES LEGAIS. (CO) 20 (VINTE) ANOS CONTADOS A PARTIR DE 25/06/2012, OBSERVADAS AS CONDICOES LEGAIS |
    优先权:
    申请号 | 申请日 | 专利标题
    US201161500543P| true| 2011-06-23|2011-06-23|
    US61/500,543|2011-06-23|
    PCT/EP2012/062240|WO2012175739A1|2011-06-23|2012-06-25|Composition comprising chitosan, glucosamine and amino acids for agricultural use|
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