WO2018183491A1 - Récupération de bactéries séchées - Google Patents

Récupération de bactéries séchées Download PDF

Info

Publication number
WO2018183491A1
WO2018183491A1 PCT/US2018/024836 US2018024836W WO2018183491A1 WO 2018183491 A1 WO2018183491 A1 WO 2018183491A1 US 2018024836 W US2018024836 W US 2018024836W WO 2018183491 A1 WO2018183491 A1 WO 2018183491A1
Authority
WO
WIPO (PCT)
Prior art keywords
bacteria
humic acid
dried
present disclosure
sample
Prior art date
Application number
PCT/US2018/024836
Other languages
English (en)
Inventor
Adrienne HUNTRESS
William PASUTTI
Ben DOUGHAN
Kimberley CLARKE
Original Assignee
Novozymes Bioag A/S
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Novozymes Bioag A/S filed Critical Novozymes Bioag A/S
Publication of WO2018183491A1 publication Critical patent/WO2018183491A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/045—Culture media therefor

Definitions

  • CFU colony forming unit
  • non-spore-forming bacteria that are in samples that are dried, dehydrated, or desiccated can be cultivated more efficiently using media that contains humic acid, salts thereof, analogs thereof, or peat, as compared to media that does not contain humic acids or related substances.
  • bacteria in spray dried samples form a greater number of colonies in CFU assays when media containing humic acid or related substances is used, as compared to the number of colonies obtained using the same media that does not contain humic acid or related substances. Therefore, for bacteria that have been exposed to drying conditions, viability assays that use bacterial growth as a threshold, provide better estimates of the number of viable cells when humic acid or related substances are included in the culture medium.
  • agar means a gelatinous substance, generally derived from seaweed, and used in culture media to provide media that is solid or semisolid in consistency. In some examples, agar concentrations of about 0.5-1.5%) (weight/volume) in media may be used for bacterial culture plates. Herein, agar is considered a type of gelling agent.
  • agrochemical means chemicals used in agriculture like, for example, chemicals used as acaricides, fungicides, gastropodicide, herbicides, insecticides, miticides, and the like.
  • an "analog" of a first substance refers to a second substance that is structurally similar to the first substance, but with some differences.
  • An analog may be synthetic.
  • an "assay” means a test to determine something.
  • bacteria means prokaryotic organisms that have peptidoglycan in their cell walls, and have lipids in their membranes, where the lipids contain fatty acids.
  • capable refers to the ability or capacity to do or achieve a specific thing.
  • colony means a visible cluster of bacteria, generally on the surface of a solid or semisolid medium (e.g., medium containing agar), and probably originating from division of a single cell.
  • a colony formed by bacteria may be called a “bacterial colony” or “colony-forming unit” (CFU).
  • sample means to have or hold.
  • something e.g., bacteria
  • count when used as a verb, means to tally or total.
  • Countering is an act to tally or total.
  • determination means to establish or find out.
  • Determining is an act to establish or find out. Something that has been established or found out may be said to be “determined.”
  • dextrose equivalent value is a measure of the amount of reducing sugars present in a sugar product.
  • the dextrose equivalent value is an indication of the average degree of polymerization for sugars.
  • dilution when used as a noun, refers to a liquid that contains a reduced concentration of a thing as compared to the liquid when undiluted. "Diluting” is an act to create a dilution.
  • solvent when referring to a solid substance, refers to the solid incorporated into a liquid so as to form a solution.
  • dried sample refers to a sample that has been treated to decrease the moisture content of the sample.
  • the bacteria may be referred to as “dried bacteria.” Dehydrated or desiccated may be used in place of the word “dried.”
  • gelling agent refers to substances that are added to liquid to cause the liquid to become solid or semisolid in consistency. A variety of these substances exist.
  • Example gelling agents may include agar, agarose, alginic acid, carrageenan, gelatin, gellan gum, guar gum, xanthan gum, and the like.
  • Gram-negative refers to bacteria that, in a Gram staining reaction, lose the crystal violet stain and take the color of the counterstain.
  • humic acid refers to a principal component of humic substances (fulvic acid and humin are other principal components of humic substances) that is soluble in dilute alkali but which becomes insoluble as the pH becomes acidic.
  • Substances "related to" humic acid may include salts of humic acid, humic acid analogs, synthetic humic acids, and may also include peat.
  • maltodextrin refers to a dextrin containing maltose.
  • medium refers to compositions for supporting growth of bacteria.
  • Example growth medium may include liquid media (e.g., broths) or solid/semisolid media (e.g., agar-containing media).
  • moisture content means the amount of water in a sample.
  • moisture content is determined on a wet basis (i.e., weight of water in a sample/total weight of sample). For example, a sample with weight 10 grams, 1 gram of which is water, has a moisture content of 0.1 or 10%.
  • non-spore forming refers to bacteria that are not capable of forming spores.
  • peat generally refers to partially decomposed vegetable/plant matter.
  • plaque refers to applying a sample, bacteria from a sample, or dilution of the sample or bacteria, to solid or semisolid bacterial culture medium (e.g., agar- containing medium).
  • solid or semisolid bacterial culture medium e.g., agar- containing medium.
  • Plated refers to something that has been applied to solid or semisolid bacterial culture medium.
  • salt refers to an ionic form of a substance.
  • sample refers to a representative part of a whole.
  • soluble means able to be dissolved (e.g., in water). “Solubilizing” is an act to dissolve something.
  • spray drying refers to processes that produce dry powders from a liquid or slurry.
  • Spray dried when referring to a substance, refers to a substance that has undergone the spray drying process.
  • synthetic refers to something that is synthesized, rather than naturally occurring.
  • a synthetic substance may be an analog.
  • unculturable when referring to a bacterium, means unable to be cultured, using current technologies (i.e., technologies prior to this disclosure; e.g., without humic acid), and generally refers to a certain set of growth conditions (e.g., the medium does not contain humic acid).
  • a bacterium that is considered unculturable may eventually be cultured, for example, when technologies are improved (e.g., with humic acid).
  • use means to employ or put into service. "Using” is an act to employ or put into service. Something that has been employed or put into service may be said to be “used.”
  • Samples (generally liquid, but also semi-solid or solid) containing bacteria may be dried, dehydrated or desiccated using a variety of methods.
  • a liquid sample may be left open so that moisture from the sample is evaporated into the air. This may be called air drying.
  • a gas stream e.g., air
  • vacuum drying where heat is supplied to the sample by conduction, radiation, or microwaves, vapor is produced and carried away by a vacuum system, may be used.
  • drum drying where a surface supplies heat to the sample, vapor is produced and carried away by an aspirator, may be used.
  • Freeze drying or lyophilization may be used in some examples and occurs when the sample is frozen and moisture from the sample is sublimed, generally under a vacuum.
  • a dried sample may be produced by draining (e.g., centrifugation to mechanically extract a solvent).
  • Spray drying may be used in some examples. Spray drying generally produces small liquid droplets of specific sizes from a liquid or slurry using a spray nozzle or atomizer. After the droplets exit the nozzle or atomizer, they are dried, generally using hot air, to form a powder. Machines known as spray dryers are normally used for this process.
  • the dry compositions may have a moisture content of less than about 50%, 40%, 30%, 25%, 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.
  • Drying of samples containing bacteria generally is considered a stress for the bacteria.
  • a variety of stresses may cause bacteria that are culturable to enter an unculturable state.
  • hydration or rehydration of bacteria in a dried, dehydrated, or desiccated sample may be a stress that may cause unculturability.
  • the amount of stress applied to the bacteria may have to be considered. For example, too much stress applied to bacteria (e.g., type of stress, time and/or intensity of the stress) may cause the bacteria to become nonviable and, therefore, not recoverable. Too little stress may fail to place bacteria into an unculturable state at all. There likely is an amount of each different type of stress that places the maximum number of bacteria in a population into an unculturable state. This amount of stress may have to be empirically determined.
  • viable but unculturable bacteria in a dried sample may become nonviable depending on the time the dried sample is stored (e.g., increasing numbers of viable, unculturable bacteria in the sample may die the longer the bacteria are exposed to the dry state) and/or depending on the conditions (e.g., temperature, relative humidity, and the like) under which the dried sample is maintained.
  • the percentage of bacteria within a sample that have entered into an unculturable state may affect the ability of that population to demonstrate recovery (e.g., if fewer bacteria in a population are in an unculturable state, assays that detect recovery of unculturable bacteria to a culturable state, even if robust, may not detect recovery).
  • an assay that can efficiently detect recovery of unculturable bacteria e.g., humic acid in the medium
  • bacteria are known to be capable of entering/exi sting in an unculturable state.
  • unculturable bacteria that exist in an unculturable state may include ⁇ -proteobacteria, ⁇ -proteobacteria, a-proteobacteria, ⁇ -proteobacteria, bacteroidetes, acinobacteria, or firmicutes.
  • the bacteria capable of entering/exi sting in an unculturable state include Gram-negative bacteria.
  • Nonlimiting examples of unculturable bacteria that exist in an unculturable state may be from the genera Acetobacter, Acinetobacter, Aeromonas, Agrobacterium, Alcaligenes, Arcobacter, Bifidobacterium, Bradyrhizobium, Burkholderia, Campylobacter, Citrobacter, Cytophaga, Enterobacter , Enter ococcus, Erwinia, Escherichia, Francisella, Helicobacter, Klebsiella, Lactobacillus, Legionella, Listeria, Oenococcus, Paracoccus, Pasteurella,
  • Nonlimiting examples of unculturable bacteria that exist in an unculturable state may be Acetobacter aceti, Acinetobacter calcoaceticus, Aeromonas hydrophilia, Aeromonas salmonicida, Agrobacterium tumifaciens, Alcaligenes eutrophus, Arcobacter butzleri,
  • Bifidobacterium lactis Bifidobacterium longum, Bifidobacteriumanimalis, Bradyrhizobium japonicum, Bradyrhizobium elkaii, Burkholderia cepacia, Burkholderia pseudomallei,
  • Campylobacter coli Campylobacter jejuni, Campylobacter lari, Citrobacter freundii, Cytophaga allerginae, Enterobacter aerogenes, Enterobacter cloacae, Enterobacter agglomerans,
  • Enterococcus faecalis Enterococcus hirae
  • Enterococcus faecium Enterococcus faecium
  • Erwinia amylovora Enterococcus faecalis, Enterococcus hirae, Enterococcus faecium, Erwinia amylovora,
  • Escherichia coli Francisella tularensis, Helicobacter pylori, Klebsiella aerogenes, Klebsiella pneumoniae, Klebsiella planticola, Lactobacillus plantarum, Lactobacillus lindneri,
  • Lactobacillus paracollinoides Lactobacillus lactus, Legionella pneumophila, Listeria monocyhtogenes, Oenococcus oeni, Paracoccus pantotrophus, Pasteurella piscicida,
  • a sample of bacteria formulated as a liquid may be dried by any of the techniques described herein. Generally, any type of formulation of the bacteria may be dried and at least some of the viable bacteria in the sample then cultured with the disclosed methods that use humic acid or humic acid related substances.
  • the liquid bacterial formulations, or the dried samples may contain any amount of bacteria. In some examples, the amount of bacteria in the formulations may be from 0.1-99% by weight.
  • the amount of bacteria in the formulations may be about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% by weight.
  • the liquid bacterial formulations, or the dried samples may contain 1 x 10 1 to about 1 x 10 20 bacteria per milliliter or gram of the formulation.
  • about 1 x 10 4 , 1 x 10 5 , 1 x 10 6 , 1 x 10 7 , 1 x 10 8 , 1 x 10 9 , 1 x 10 10 , 1 x 10 11 , 1 x 10 12 , 1 x 10 13 , 1 x 10 14 , 1 x 10 15 , or more bacteria or CFU per milliliter or gram of the formulation may be present.
  • the liquid bacterial formulations, or the dried samples may contain non-aqueous carriers.
  • non-aqueous carriers include disaccharides, maltodextrins, monosaccharides, oligosaccharides, sugar alcohols, peat-based powders and granules, agriculturally acceptable polymers, and the like.
  • the non-aqueous carriers may constitute any suitable portion of the liquid bacterial formulations, or the dried samples.
  • the non-aqueous carrier(s) comprise(s) about 1 to about 99 % (by weight) of the compositions.
  • the non-aqueous carrier(s) constitute(s) about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
  • the carrier amount/concentration is about 5 to about 45%, about 10 to about 30%, about 50 to about 99%, about 55% to about 95%, about 60% to about 95%, about 65% to about 90%, about 70 to about 90%, about 75 to about 90%, about 80 to about 90% or about 80 to about 85% (by weight) of the composition.
  • liquid bacterial formulations or the dried samples of the present disclosure, comprise one or more commercial carriers used in accordance with the manufacturer's recommended amounts/concentrations.
  • the non-aqueous carrier comprises, consists essentially of, or consists of one or more maltodextrins.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable maltodextrins, including, but not limited to, maltodextrins having a dextrose equivalent value (DEV) of about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25.
  • compositions of the present disclosure comprise one or more maltodextrins having a DEV of about 5 to about 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25, about 10 to about 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25, or about 15 to about 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25.
  • the non-aqueous carrier comprises a combination of maltodextrins having a DEV of about 5 to about 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25, about 10 to about 11,
  • the non-aqueous carrier comprises one or more maltodextrins having a DEV of about 10 to about 25 (e.g., one or more maltodextrins having a DEV of about 15 to about 20). In some embodiments, the non-aqueous carrier comprises a combination of maltodextrins having a DEV of about 10 to about 25 (e.g., a combination of maltodextrins having a DEV of about 15 to about 20).
  • Maltodextrins may be incorporated into the liquid bacterial formulations, or the dried samples of the present disclosure, in any suitable amount(s)/concentration(s). In some
  • the maltodextrin(s) comprise about 5 to about 99 % or more (by weight) of the composition.
  • the non-aqueous carrier comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99% or more of one or more maltodextrins (each and/or collectively) having a DEV value of about 15 to about 20.
  • the maltodextrin amount/concentration is about 50 to about 95%, about 55% to about 90%, about 60% to about 85%, about 65% to about 80%, or about 70 to about 80% (by weight) of the non-aqueous carrier.
  • the non-aqueous carrier comprises, consists essentially of, or consists of one or more monosaccharides, disaccharides, sugar alcohols and/or oligosaccharides.
  • concentration of these substances in the compositions disclosed herein may be between about 1-90%, 2-80%, 3-70%, 4-60%, 5-50%, 5-40%, 5-30%, 5-20% or 6-15% (by weight).
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable monosaccharide, including, but limited to, allose, altrose, arabinose, fructose, galactose, glucose, gulose, iodose, lyxose, mannose, ribose, talose, threose and xylose.
  • the non-aqueous carrier comprises glucose. In some embodiments, the non-aqueous carrier does not comprise glucose.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable disaccharide, including, but limited to, cellobiose, chitobiose, gentiobiose, gentiobiulose, isomaltose, kojibiose, lactose, lactulose, laminaribiose, maltose (e.g., maltose monohydrate, anhydrous maltose), maltulose, mannobiose, melibiose, melibiulose, nigerose, palatinose, rutinose, rutinulose, sophorose, sucrose, trehalose, turanose and xylobiose.
  • maltose e.g., maltose monohydrate, anhydrous maltose
  • maltulose e.g., maltose monohydrate, anhydrous maltose
  • maltulose e.g., maltose monohydrate
  • the non-aqueous carrier comprises maltose. In some embodiments, the nonaqueous carrier comprises sucrose. In some embodiments, the non-aqueous carrier comprises trehalose. In some embodiments, the non-aqueous carrier does not comprise trehalose.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable oligosaccharide, including, but limited to, fructo-oligosaccharides, galacto-oligosaccharides, mannon-oligosaccharides and raffinose.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable sugar alcohol, non-limiting examples of which include sorbitol, xylitol, glycerol, erythritol, threitol, arabitol, ribitol, mannitol, galactitol, fucitol, iditol, inositol, volemitol, isomalt, maltitol, lactitol, maltotriitol, maltotetraitol and polyglycitol.
  • suitable sugar alcohol non-limiting examples of which include sorbitol, xylitol, glycerol, erythritol, threitol, arabitol, ribitol, mannitol, galactitol, fucitol, iditol, inositol, volemitol, isomalt, mal
  • Mono-, di- and oligosaccharides may be incorporated into the liquid bacterial formulations, or the dried samples of the present disclosure, in any suitable
  • the mono-, di- and/or oligosaccharide(s) comprise(s) about 5 to about 95% (by weight) of the composition.
  • the non-aqueous carrier comprises about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95% or more (by weight) of one or more mono-, di- and/or oligosaccharides.
  • the mono-, di- and/or oligosaccharide amount/concentration is about 1 to about 65%, about 5% to about 20%, about 10% to about 25%, about 20% to about 50%, or about 30 to about 60% (by weight) of the nonaqueous carrier.
  • the non-aqueous carrier comprises, consists essentially of, or consists of one or more malt extracts.
  • the non-aqueous carrier comprises, consists essentially of, or consists of one or more peat-based powders and/or granules.
  • liquid bacterial formulations may comprise any suitable peat-based powder(s) and/or granule(s).
  • Peat-based powders and/or granules may be incorporated into the compositions of the present disclosure in any suitable form.
  • Peat-based powders and/or granules may be incorporated into the liquid bacterial formulations, or the dried samples of the present disclosure in any suitable
  • the peat-based powder(s) and/or granuale(s) comprise(s) about 5 to about 95 % (by weight) of the composition.
  • the non-aqueous carrier comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95% or more (by weight) of one or more peat-based powders and/or granuales.
  • the peat extract amount/concentration is about 50 to about 99%, about 55% to about 95%, about 60% to about 90%, about 65% to about 90%, or about 70 to about 90% (by weight) of the non-aqueous carrier.
  • the non-aqueous carrier comprises, consists essentially of, or consists of one or more agriculturally acceptable polymers.
  • compositions of the present disclosure may comprise any suitable agriculturally acceptable polymer(s), including, but not limited to, biodegradable polymers and synthetic polymers.
  • suitable agriculturally acceptable polymer(s) including, but not limited to, biodegradable polymers and synthetic polymers.
  • compositions of the present disclosure comprise agar, alginate, carrageenan, cellulose, guar gum, locust bean gum, methylcellulose, pectin, polycaprolactone, polylactide, polyvinyl alcohol, polyvinyl pyrrolidone, sodium carboxymethyl cellulose, starch and xanthan gum.
  • Non-limiting examples of polymers that may be useful in compositions of the present disclosure include TICAXAN® xanthan powders, such as PRE-HYDRATED® TICAXAN® Rapid-3 Powder (TIC Gums, White Marsh, MD) and combinations thereof.
  • compositions of the present disclosure may be found in Pouci, et al. AM. J. AGRIC. BIOL. SCI. 3(1):299 (2008).
  • Agriculturally acceptable polymers may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the agriculturally acceptable polymer(s) comprise(s) about 5 to about 95 % (by weight) of the composition.
  • the non-aqueous carrier comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95% or more (by weight) of one or more agriculturally acceptable polymers.
  • the agriculturally acceptable polymers amount/concentration is about 50 to about 99%, about 55% to about 95%, about 60% to about 90%), about 65%> to about 90%>, or about 70 to about 90%> (by weight) of the nonaqueous carrier.
  • liquid bacterial formulations or the dried samples of the present disclosure, comprise one or more commercial polymers used in accordance with the manufacturer's recommended amounts/concentrations.
  • compositions of the present disclosure comprise a dry non-aqueous carrier that comprises one or more maltodextrins in combination with one or more mono-, di- and/or oligosaccharides, one or more sugar alcohols, one or more malt extracts, one or more
  • Maltodextins and mono-, di- and/or oligosaccharides may be incorporated into compositions of the present disclosure in any suitable ratio(s).
  • the nonaqueous carrier has a maltodextrin:(mono-, di- and/or oligosaccharide) (e.g.,
  • the non-aqueous carrier has a
  • maltodextrin:(mono-, di- and/or oligosaccharide) e.g., maltodextrin: maltose
  • ratio of about 5:95, 10:90, 15:85, 20:80, 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 67:33, 70:30, 75:25, 80:20, 85: 15, 90: 10, 95:5 or more.
  • the maltodextrin: (mono-, di- and/or oligosaccharide) e.g., maltodextrimmaltose
  • the compositions of the present disclosure comprise one or more
  • maltodextrins having a DEV of about 15 to about 20 and one or more mono-, di- and/or oligosaccharides (e.g., maltose) in a ratio of about 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85: 15, 90: 10, 95:5.
  • mono-, di- and/or oligosaccharides e.g., maltose
  • Maltodextins and malt extracts may be incorporated into the liquid bacterial formulations, or the dried samples of the present disclosure, in any suitable ratio(s).
  • the non-aqueous carrier has a maltodextrin: malt extract ratio of about 1 :99 to about 99: 1 (by weight, based upon the respective weight percentages of the maltodextrin(s) and malt extract(s) in the non-aqueous carrier).
  • the non-aqueous carrier has a maltodextrin: malt extract ratio of about 5:95, 10:90, 15:85, 20:80, 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85: 15, 90: 10, 95:5 or more.
  • the maltodextrin: malt extract ratio is about 45:55 to about 95:5.
  • the compositions of the present disclosure comprise one or more
  • maltodextrins having a DEV of about 15 to about 20 and one or more malt extracts in a ratio of about 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85: 15, 90: 10, 95:5.
  • Maltodextins and peat-based powders and/or granules may be incorporated into the liquid bacterial formulations, or the dried samples of the present disclosure, in any suitable ratio(s).
  • the non-aqueous carrier has a maltodextrin: peat powder/granuale ratio of about 1 : 99 to about 99: 1 (by weight, based upon the respective weight percentages of the maltodextrin(s) and peat powder(s)/granuale(s) in the non-aqueous carrier).
  • the non-aqueous carrier has a maltodextrin: peat powder/granuale ratio of about 5:95, 10:90, 15:85, 20:80, 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85: 15, 90: 10, 95:5 or more.
  • the maltodextrin: peat powder/granuale ratio is about 45:55 to about 95:5.
  • compositions of the present disclosure comprise one or more maltodextrins having a DEV of about 15 to about 20 and one or more peat-based powders and/or granuales in a ratio of about 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85: 15, 90: 10, 95:5.
  • Non-aqueous carriers may be incorporated into the liquid bacterial formulations, or the dried samples of the present disclosure, in any suitable amount(s)/concentration(s).
  • the non-aqueous carrier(s) comprise(s) about 5 to about 99.9% (by weight) of the composition.
  • the non-aqueous carrier(s) constitute(s) about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 99.5%) or more (by weight) of the composition.
  • the non-aqueous carrier amount/concentration is about 50 to about 99%, about 55% to about 95%, about 60%> to about 95%, about 65% to about 90%, about 70 to about 90%, about 75 to about 90%, about 80 to about 90%) or about 80 to about 85%> (by weight) of the composition.
  • liquid bacterial formulations or the dried samples of the present disclosure, comprise one or more commercial carriers used in accordance with the manufacturer's recommended amounts/concentrations.
  • liquid bacterial formulations may comprise any suitable agriculturally acceptable dispersant(s), including, but not limited to, surfactants and wetting agents.
  • liquid bacterial formulations or the dried samples of the present disclosure, comprise one or more anionic surfactants.
  • anionic surfactants for example, in some embodiments, in some
  • compositions of the present disclosure comprise one or more water-soluble anionic surfactants and/or one or more water-insoluble anionic surfactants, optionally one or more anionic surfactants selected from the group consisting of alkyl carboxylates (e.g., sodium stearate), alkyl sulfates (e.g., alkyl lauryl sulfate, sodium lauryl sulfate), alkyl ether sulfates, alkyl amido ether sulfates, alkyl aryl polyether sulfates, alkyl aryl sulfates, alkyl aryl sulfonates, alkyl sulfonates, alkyl amide sulfonates, alkyl aryl sulfonates, alkyl benzene sulfonates, alkyl diphenyloxide sulfonate, alpha-olefin sulfonates, alkyl
  • perfluorooctanesulfonate perfluorooctanesulfonate, phosphate ester, styrene acrylic polymers, toluene sulfonates and xylene sulfonates.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more cationic surfactants.
  • one or more cationic surfactants for example, in some embodiments, in some
  • compositions of the present disclosure comprise one or more pH-dependent amines and/or one or more quaternary ammonium cations, optionally one or more cationic surfactants selected from the group consisting of alkyl trimethyl ammonium salts (e.g., cetyl trimethylammonium bromide, cetyl trimethylammonium chloride), cetylpyridinium chloride, benzalkonium chloride, benzethonium chloride, 5-Bromo-5-nitro-l,3-dioxane,
  • alkyl trimethyl ammonium salts e.g., cetyl trimethylammonium bromide, cetyl trimethylammonium chloride
  • cetylpyridinium chloride e.g., cetyl trimethylammonium bromide, cetyl trimethylammonium chloride
  • cetylpyridinium chloride e.g., benzalkonium chloride, benzethonium chloride, 5-Bromo-5
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more nonionic surfactants.
  • nonionic surfactants for example, in some embodiments, in some
  • compositions of the present disclosure comprise one or more water-soluble nonionic surfactants and/or one or more water-insoluble nonionic surfactants, optionally one or more nonionic surfactants selected from the group consisting of alcohol ethoxylates (e.g., TERGITOLTM 15-S surfactants, such as TERGITOLTM15-S-9 (The Dow Chemical
  • alkanolamides alkanolamine condensates, carboxylic acid esters, cetostearyl alcohol, cetyl alcohol, cocamide DEA, dodecyldimethylamine oxides, ethanolamides, ethoxylates of glycerol ester and glycol esters, ethylene oxide polymers, ethylene oxide- propylene oxide copolymers, glucoside alkyl ethers, glycerol alkyl ethers (e.g., ), glycerol esters, glycol alkyl ethers (e.g., polyoxyethylene glycol alkyl ethers, polyoxypropylene glycol alkyl ethers,), glycol alkylphenol ethers (e.g., polyoxyethylene glycol alkylphenol ethers,), glycol esters, monolaurin, pentaethylene glycol monododecyl ethers, poloxamer, polyamines, polyg
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise at least one nonionic surfactant.
  • the compositions of the present disclosure comprise at least one water insoluble nonionic surfactant and at least one water soluble nonionic surfactant.
  • the compositions of the present disclosure comprise a combination of nonionic surfactants having hydrocarbon chains of substantially the same length.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more zwitterionic surfactants.
  • the compositions of the present disclosure comprise one or more betaines and/or one or more sultaines, optionally one or more zwitterionic surfactants selected from the group consisting of 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfonate, cocamidopropyl betaine, cocamidopropyl hydroxysultaine, phosphatidylserine, phosphatidylethanolamine, phosphatidylcholine and/or one or more sphingomyelins.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more soaps and/or organosilicone surfactants.
  • the compositions of the present disclosure comprise one or more alkali metal salts of fatty acids.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more wetting agents.
  • the compositions of the present disclosure comprise one or more naphthalene sulfonates, optionally one or more alkyl naphthalene sulfonates (e.g., sodium alkyl naphthalene sulfonate), one or more isopropyl naphthalene sulfonates (e.g., sodium isopropyl naphthalene sulfonate) and/or one or more butyl naphthalene sulfonates (e.g., sodium n-butyl naphthalene sulfonate).
  • alkyl naphthalene sulfonates e.g., sodium alkyl naphthalene sulfonate
  • isopropyl naphthalene sulfonates e.g., sodium isopropyl
  • surfactant(s) will have low toxicity for the bacteria in the composition and for the plant part(s) to which the composition is to be applied. In some embodiments, the surfactant(s) will be selected to wet and/or emulsify one or more soils.
  • Non-limiting examples of dispersants that may be useful in compositions of the present disclosure include AtloxTM (e.g., 4916, 4991; Croda International PLC, Edison, NJ), BIO-SOFT® (e.g., N series, such as Nl-3, Nl-7, Nl-5, Nl-9, N23-3, N2.3-6.5, N25-3, N25-7, N25-9, N91-2.5, N91-6, N91-8; Stepan Company, Northfield, IL), MAKON® nonionic surfactants (e.g., DA-4, DA-6 and DA-9; Stepan Company, Northfield, IL), MORWET® powders (Akzo Nobel Surface Chemistry LLC, Chicago, IL), MULTIWETTM surfactants (e.g., MO-70R, MO-85P, MO-85P-PW-(AP); Croda International PLC, Edison, NJ), SILWET® L-77 (Helena Chemical Company, Collierville, TN
  • Dispersants may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the dispersant(s) comprise(s) about 0.1 to about 25 % (by weight) of the composition.
  • the compositions of the present disclosure may comprise about 0.1, 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 15, 20, 25% or more (by weight) of one or more dispersants.
  • the dispersant (s) comprise(s) about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1 to about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 % (by weight) of the composition.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more commercial wetting agents and/or one or more surfactants used in accordance with the manufacturer's recommended amounts/concentrations.
  • liquid bacterial formulations may comprise any suitable excipient(s), including, but not limited to, anti-freezing agents, drying agents, safeners and pH buffers.
  • liquid bacterial formulations may comprise any suitable agriculturally acceptable anti-freezing agent(s), including, but not limited to, ethylene glycol, glycerin, propylene glycol and urea.
  • suitable agriculturally acceptable anti-freezing agent(s) including, but not limited to, ethylene glycol, glycerin, propylene glycol and urea.
  • compositions of the present disclosure comprise one or more commercial anti-freezing agents used in accordance with the manufacturer's recommended amounts/concentrations.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable agriculturally acceptable drying agent(s), including, but not limited to, drying powders.
  • the compositions of the present disclosure comprise calcium stearate, clay (e.g., attapulgite clay, montmorillonite clay), graphite, magnesium stearate, magnesium sulfate, powdered milk, silica (e.g., fumed silica,
  • hydrophobically-coated silica precipitated silica
  • soy lecithin hydrophobically-coated silica, precipitated silica), soy lecithin and/or talc.
  • Non-limiting examples of drying agents that may be useful in compositions of the present disclosure include AEROSIL® hydrophobic fumed silica powders (Evonik Corporation, Parsippany, NJ), BENTOLITE® powders (BYK-Chemie GmbH, Wesel, Germany), SIPERNAT® silica powders (Evonik Corporation, Parsippany, NJ) and combinations thereof.
  • drying agents that may be included in the compositions of the present disclosure may be found in BURGES, FORMULATION OF MICROBIAL BIOPESTICIDES:
  • Drying agents may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the compositions of the present disclosure comprise about 0.5 to about 10 grams of drying powder per liter of composition.
  • the compositions of the present disclosure may comprise about 0.5, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10 grams or more of drying powder per liter of the composition.
  • the amount/concentration of drying agent(s) comprise(s) calcium stearate, attapulgite clay, montmorillonite clay, graphite, magnesium stearate, silica (e.g., fumed silica, hydrophobically- coated silica and/or precipitated silica) and/or talc.
  • liquid bacterial formulations or the dried samples of the present disclosure, comprise one or more commercial drying agents used in accordance with the manufacturer's recommended amounts/concentrations.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable agriculturally acceptable dust suppressant(s), including, but not limited to, adhesives, glycerin, mineral oils, paraffinic oils, vegetable oils and synthetic polymers. It is to be understood that some compounds may act as both an adhesive and a dust suppressant. Indeed, the dust suppressant activity of many compounds arises from their ability to adhere dust particles to other heavier particles. For example, in some embodiments of the present disclosure, one or more oils is used to adhere microbial spores in the composition to larger particles comprising one or more maltodextrins.
  • Non-limiting examples of dust suppressants that may be useful in compositions of the present disclosure include ARENAPRO, BIORAIN and ROADKILL (Dustkill LLC, Columbus, IN), BIO-SOFT® surfactants (e.g., N series, such as Nl-3, Nl-7, Nl-5, Nl-9, N23-3, N2.3-6.5, N25-3, N25-7, N25-9, N91-2.5, N91-6, N91-8; Stepan Company, Northfield, IL), DURASOIL (Soilworks, LLC, Scottsdale, AZ), DUSGON (Dupont (Australia) Ltd, Macquarie Park,
  • BIO-SOFT® surfactants e.g., N series, such as Nl-3, Nl-7, Nl-5, Nl-9, N23-3, N2.3-6.5, N25-3, N25-7, N25-9, N91-2.5, N91-6, N91-8; Stepan Company, Northfield, IL), DURASOIL (Soilworks, LLC, Scottsdale,
  • SUNSPRAY® ULTRA-FINE® oils HollyFrontier Refining & Marketing, LLC, Plymouth Meeting, PA
  • TOMADOL® surfactants e.g., 23-1, 23-3, 23-5, 23-6.5; Air Products and Chemicals, Inc., Allentown, PA
  • Dust suppressants may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • dust suppressants comprise about 0.1 to about 30% (by weight) of the composition.
  • the compositions of the present disclosure may comprise about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 6, 7, 8, 9, 10% or more (by weight) of one or more dust suppressants.
  • the dust suppressant amount/concentration is about 1 to about 2, 3, 4, 5, 6, 7, 8, 9 or 10%, about 1.5 to about 2, 3, 4, 5, 6, 7, 8, 9 or 10%, about 2 to about 3, 4, 5, 6, 7, 8, 9 or 10%, about 2.5 to about 3, 4, 5, 6, 7, 8, 9 or 10% or about 3 to about 4, 5, 6, 7, 8, 9 or 10%) (by weight) of the inculant composition.
  • compositions of the present disclosure comprise one or more surfactants, such as BIO-SOFT® N23-3, that disperse microorganisms within the composition and suppress dusting.
  • the non-aqueous carrier in the compositions of the present invention may act as a protectant. It is to understood that the compositions of the present disclosure may comprise additional protectants.
  • Protectants may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the protectant(s) comprise(s) about 0.1 to about 25 % (by weight) of the composition.
  • the compositions of the present disclosure may comprise about 0.1, 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 15, 20, 25% or more (by weight) of one or more protectants.
  • the protectant amount/concentration is about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1 to about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 % (by weight) of the
  • the protectant amount/concentration is effective to ensure that at least about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65% or more of the microbial spores/vegetative cells in the composition survive following storage at 10,
  • the protectant amount/concentration is effective to ensure that at least about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65% or more of the microbial spores/vegetative cells in the composition survive following desiccation (of about 0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95% or more) and storage at 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85% or more relative humidity for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 21, 22, 23, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104 weeks or more.
  • the protectant amount/concentration is effective to ensure that at least about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65% or more of the microbial spores/vegetative cells in the composition survive following
  • cryopreservation at or below -80°C for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
  • compositions of the present disclosure compriviate
  • sugar alcohols e.g., sorbitol
  • Sugar alcohols may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s). In some embodiments, the sugar alcohol(s)
  • compositions of the present disclosure may comprise about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.75, 1, 1.25, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5% or more (by weight) of one or more sugar alcohols (e.g., sorbitol).
  • sugar alcohols e.g., sorbitol
  • liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more peat extracts.
  • Peat extracts may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the peat extract(s) comprise(s) about 0.1 to about 25 % (by weight) of the composition.
  • the compositions of the present disclosure may comprise about 0.1, 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 15, 20, 25% or more (by weight) of one or more peat extracts.
  • the peat extract amount/concentration is about 0.5 to about 10% (by weight) of the the composition.
  • compositions of the present disclosure comprise one or more peptones.
  • liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more skim milk extracts.
  • skim milk extracts may be incorporated into the compositions of the present disclosure in any suitable amount/concentration.
  • skim milk extracts comprise about 0.1 to about 25 %> (by weight) of the the composition.
  • the compositions of the present disclosure may comprise about 0.1, 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 15, 20, 25% or more (by weight) of skim milk extracts.
  • the skim milk extract amount/concentration is about 0.5 to about 10%) (by weight) of the composition.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more hygroscopic polymers.
  • the compositions of the present disclosure may comprise one or more albumins, alginates, celluloses, gums (e.g., cellulose gum, guar gum, gum arabic, gum combretum, xantham gum), methyl celluloses, nylons, pectins, polyacrylic acids, polycarbonates, polyethylene glycols (PEG),
  • PEI polyethylenimines
  • PMA polymethylacrylates
  • PVA polyurethanes
  • PVA polyvinyl alcohols
  • PVP polyvinylpyrrolidones
  • propylene glycols sodium carboxymethyl celluloses and/or starches.
  • Non-limiting examples of hygroscopic polymers that may be useful in compositions of the present disclosure include AGRIMERTM polymers (e.g., 30, AL-10 LC, AL-22, AT/ATF, VA 3E, VA 31, VA 5E, VA 51, VA 6, VA 6E, VA 7E, VA 71, VEMA AN-216, VEMA AN-990, VEMA AN-1200, VEMA AN-1980, VEMA H-815MS; Ashland Specialty Ingredients,
  • compositions of the present disclosure may be found in Pouci, et al. AM. J. AGRIC. BIOL. SCI. 3(1):299 (2008).
  • Hygroscopic polymers may be incorporated into the liquid bacterial formulations, or the dried samples of the present disclosure in any suitable amount(s)/concentration(s).
  • the hygroscopic polymer(s) comprise(s) about 0.1 to about 25 % (by weight) of the composition.
  • the hygroscopic polymer(s) comprise(s) about 0.5 to about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 % (by weight) of the composition.
  • the compositions of the present disclosure may comprise about 0.1, 0.2, 0.3, 0.4, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 15, 20, 25% or more (by weight) of one or more hygroscopic polymers.
  • the hygroscopic polymer amount/concentration is about 0.5 to about 10% (by weight) of the composition.
  • the hygroscopic polymer amount/concentration is about 0.5 to about 5% (by weight) of the composition.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more commercial hygroscopic polymers used in accordance with the manufacturer's recommended amounts/concentrations.
  • the liquid bacterial formulations, or the dried samples of the present disclosure comprise one or more oxidation control components.
  • compositions of the present disclosure may comprise any suitable oxidation control component(s), including, but not limited to, antioxidants and/or oxygen scavengers.
  • the oxidation control component is/comprises ascorbic acid and/or glutathione.
  • the liquid bacterial formulations, or the dried samples comprise one or more antioxidants.
  • the compositions of the present disclosure comprise ascorbic acid, ascorbyl palmitate, ascorbyl stearate, calcium ascorbate, carotenoids, lipoic acid, phenolic compounds (e.g., flavonoids, flavones, flavonols), potassium ascorbate, sodium ascorbate, thiols (e.g., glutathione, lipoic acid, N-acetyl cysteine), tocopherols, tocotrienols, ubiquinone and/or uric acid.
  • ascorbic acid ascorbyl palmitate, ascorbyl stearate, calcium ascorbate, carotenoids, lipoic acid, phenolic compounds (e.g., flavonoids, flavones, flavonols), potassium ascorbate, sodium ascorbate, thiols (e.g., glutathione, lipoic acid
  • Non-limiting examples of antioxidants that may be useful in compositions of the present disclosure include those that are soluble in the cell membrane (e.g., alpha tocopherol (vitamin E), ascorbyl palmitate), those that are soluble in alcohols (e.g., IRGANOX®
  • antioxidants BASF Buch AG, Basel, Switzerland
  • those that are soluble in water e.g., ascorbic acid and isomers or ascorbic acid, sodium or potassium salts of ascorbic acid or isomers or ascorbic acid, glutathione, sodium or potassium salts of glutathione.
  • use of a membrane-soluble antioxidant necessitates the addition of one or more surfactants to adequately disperse the antioxidant within the composition.
  • compositions of the present disclosure comprise one or more commercial antioxidants used in accordance with the manufacturer's recommended amounts/concentrations.
  • the liquid bacterial formulations, or the dried samples comprise one or more naturally occurring or synthetic oxygen scavengers.
  • the compositions of the present disclosure comprise ascorbic acid, ascorbate salts, catechol and/or sodium hydrogen carbonate.
  • compositions of the present disclosure comprise one or more commercial oxygen scavengers used in accordance with the manufacturer's recommended amounts/concentrations.
  • Oxidation control components may be incorporated into the liquid bacterial formulations, or the dried samples compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the oxidation control component(s) comprise(s) about 0.0001 to about 5 % or more (by weight) of the composition.
  • the compositions of the present disclosure may comprise about 0.0005, 0.001, 0.002, 0.003, 0.004, 0.005, 0.0075, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.75, 1, 1.25, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5% or more of one or more oxidation control components.
  • the amount/concentration of oxidation control components is about 0.005 to about 2% (by weight) of the composition. In some embodiments, the oxidation control component(s) is/are present in a concentration ranging from about 1 x 10 "20 M to about 1 x 10 "1 M.
  • one or more oxidation control components may be added at a concentration of 1 x 10 "20 M, 1 x 10 "19 M, 1 x 10 "18 M, 1 x 10 "17 M, 1 x 10 "16 M, 1 x 10 "15 M, 1 x 10 "14 M, 1 x 10 "13 M, 1 x 10 "12 M, 1 x 10 "11 M, 1 x 10 "10 M, 1 x 10 "9 M, 1 x 10 "8 M, 1 x 10 "7 M, 1 x 10 "6 M, 1 x 10 "5 M, 1 x 10 "4 M, 1 x 10 "3 M, 1 x 10 "2 M, 1 x 10 "1 M or more.
  • liquid bacterial formulations may comprise any suitable safener(s), including, but not limited to, napthalic anhydride.
  • compositions of the present disclosure may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the compositions of the present disclosure comprise one or more commercial safeners used in accordance with the manufacturer's recommended
  • liquid bacterial formulations, or the dried samples compositions of the present disclosure may comprise any suitable pH buffer(s), including, but not limited to, potassium phosphate monobasic and potassium phosphate dibasic.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable adhesive(s), including, but not limited to, adhesive compositions comprising one or more maltodextrins and/or one or more mono-, di- or oligosaccharides.
  • the compositions of the present disclosure may be formulated into any suitable type of composition, including, but not limited to, seed coatings, soil formulations and foliar formulations.
  • the liquid bacterial formulations, or the dried samples of the present disclosure may comprise any suitable anti-settling agent(s), including, but not limited to, polyvinyl acetate, polyvinyl alcohols with different degrees of hydrolysis, polyvinylpyrrolidones, polyacrylates, acrylate-, polyol- or polyester-based paint system binders which are soluble or dispersible in water, moreover copolymers of two or more monomers such as acrylic acid, methacrylic acid, itaconic acid, maleic acid, fumaric acid, maleic anhydride, vinylpyrrolidone, ethylenically unsaturated monomers such as ethylene, butadiene, isoprene, chloroprene, styrene,
  • any suitable anti-settling agent(s) including, but not limited to, polyvinyl acetate, polyvinyl alcohols with different degrees of hydrolysis, polyvinylpyrrolidones, polyacrylates, acrylate-, polyol- or
  • Anti-settling agents may be incorporated into the compositions of the present disclosure in any suitable amount(s)/concentration(s).
  • the compositions of the present disclosure comprise about 0.0001 to about 10% or more (by weight) of the composition.
  • the compositions of the present disclosure may comprise about 0.0005, 0.001, 0.002, 0.003, 0.004, 0.005, 0.0075, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.75, 1, 1.25, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10% or more of one or more anti-settling agents.
  • the amount/concentration of anti-settling agents is about 0.01 to about 5% (by weight) of the composition.
  • compositions of the present disclosure comprise one or more commercial anti-settling agents used in accordance with the manufacturer's recommended amounts/concentrations.
  • the liquid bacterial formulations, or the dried samples compositions of the present disclosure may comprise any suitable biostimulant(s), including, but not limited to, seaweed extracts (e.g., Ascophyllum nodosum extracts, such as alginate, Ecklonia maxima extracts, etc.), humic acids (e.g., potassium humate), fulvic acids, myo-inositol, glycine and combinations thereof.
  • seaweed extracts e.g., Ascophyllum nodosum extracts, such as alginate, Ecklonia maxima extracts, etc.
  • humic acids e.g., potassium humate
  • fulvic acids e.g., myo-inositol, glycine and combinations thereof.
  • Soil organic matter may be classified as a humic substance or a non-humic substance.
  • Humic substances are composed of altered or transformed components of plants, animals, microbes, and the like (e.g., decomposed organic matter).
  • Non-humic substances include unaltered remains (e.g., not decomposed) of plants, animals, microbes, and the like.
  • Humic substances are generally thought to include a humic acid component, a fulvic acid component, and a humin component.
  • the humic acid component, and substances that may contain all or part of the humic acid component, is disclosed herein as capable of increasing the efficiency of plating of unculturable bacteria from samples.
  • humic acid component for example, is generally water soluble at alkaline pH, but becomes less soluble under acidic conditions.
  • humic acid may be defined as the fraction of humic substances that are water insoluble at pH 2, but are increasingly soluble at higher pH values.
  • the fulvic acid component is generally soluble in water at all pH values.
  • the humin component is generally insoluble at all pH values.
  • humic acid is a complex mixture of weak aliphatic and aromatic organic acids, often containing phenolic and carboxylic substituents.
  • Humic acids may be called polydisperse because of their variable chemical features.
  • the molecular sizes of humic acids may range, in some examples, from approximately about 10,000 to about 100,000 daltons.
  • Humic acids may readily form salts with inorganic trace mineral elements. Both humic acids and salts thereof can be used and may be active in the methods disclosed herein.
  • Humic substances may be components of soil (e.g., humus), peat, lignite, coal, lake and stream sediments, seawater, and shale (e.g., Leonardite).
  • Humic acid may be obtained or extracted from certain of these substances (e.g., convenient sources may be humus rich soil, peat moss, compost) using various methods.
  • Humic acid may also be obtained from systems set up to facilitate degradation of organic materials (e.g., plant material) so that humic acid is produced.
  • Humic acid may also be formed by polymerization of substances like polyphenols. Some of these methods are described in, for example, US Patent No. 5,854,032. Other methods for extracting or producing humic acids may be used.
  • Humic acids can also be purchased commercially (e.g., Sigma- Aldrich No. 53680; Alfa Aesar No. 41747).
  • the above-mentioned substances - like peat, lignite, coal, sediments, seawater, shale, and the like - are also within the scope of materials that increase plating efficiency of unculturable bacteria.
  • Salts of humic acid are within the scope of materials that can increase the efficiency of plating or recovery of unculturable bacteria from samples.
  • formation of salts of humic acid depends on the ability of carboxyl and/or hydroxyl groups therein to dissociate their hydrogen ions and bind to positive cations (e.g., metal cations like iron, copper, zinc, calcium, manganese, magnesium, and the like).
  • Salts of humic acid can be purchased commercially (Sigma-Aldrich No. H16752).
  • Humic acid analogs and synthetic humic acids (a humic acid analog may also be synthetic) also exist and are within the scope of materials that may increase the efficiency of plating of unculturable bacteria.
  • certain quinones one being anthraquinone- 2, 6-disulfonate (AQDS) are considered analogs of humic acid.
  • Synthetic humic acids can be made by methods known in the art (e.g., V. A. Litvin, R. L. Galagan. "Synthesis and Properties of Synthetic Analogs of Natural Humic Acids.” Russian Journal of Applied Chemistry 85, no. 2, 2012).
  • Humic acid may be fractionated and some of the fractions may be successfully used in the methods disclosed herein.
  • humic acid is added to an aqueous solution of 0.1 M ammonium bicarbonate at a slightly basic pH. Insoluble material is removed from the mixture. The remaining solution is passed through a filter that retains molecules larger than 5,000 molecular weight on the filter, while molecules smaller than 5,000 molecular weight pass through the filter. The material retained on the filter may be shown to possess the activity of increasing the efficiency of plating of unculturable bacteria.
  • Other methods of fractionating humic acid may be used.
  • humic acid, salts thereof, analogs thereof, and peat may include leonardite humic acids, lignite humic acids, peat humic acids or water-extracted humic acids.
  • humic acid, salts thereof, analogs thereof, and peat may include ammonium humate, boron humate, potassium humate and/or sodium humate.
  • ammonium humate, boron humate, potassium humate and sodium humate is/are excluded.
  • Nonlimiting examples of humic acids that may be useful various examples may include MDL Number MFCD00147177 (CAS Number 1415-93-6), MDL Number MFCD00135560 (CAS Number 68131-04-4), MDL Number MFCS22495372 (CAS Number 68514-28-3), CAS Number 93924-35-7, and CAS Number 308067-45-0.
  • Dried bacterial populations likely contain viable but unculturable bacteria.
  • Unculturable bacteria were generally once culturable under a specific set of conditions, but became unculturable at a later time under those same culture conditions.
  • application of stress e.g., drying
  • Recovery of unculturable bacteria may occur when the unculturable bacteria become culturable later.
  • humic acid may be added without salts, analogs, or peat.
  • salts of humic acid may be added, without humic acid, analogs, or peat.
  • analogs of humic acid may be added, without humic acid, salts thereof, or peat.
  • peat may be added, without humic acid, salts thereof, or analogs thereof.
  • the humic acids and/or related substances may be added to any bacterial medium.
  • the growth media may include YEM, R2A, TSA, LB, NA, ISP2, Jensen's, and the like.
  • Media used for culturing bacteria may be liquid, semisolid or solid.
  • Semisolid or solid medium may be made, in some examples, by adding a gelling agent to a liquid medium.
  • a common gelling agent is agar. However, a number of other gelling agents exist and may be used. Examples include agarose, alginic acid, carrageenan, gelatin, gellan gum, guar gum, xanthan gum, and others.
  • bacteria plated on a semisolid or solid medium may divide and form colonies after a time when the medium is placed in an environment conducive to growth of bacteria (e.g., 2-5 days of incubation at 30°C in an ambient atmosphere).
  • an environment conducive to growth of bacteria e.g., 2-5 days of incubation at 30°C in an ambient atmosphere.
  • these conditions e.g., days of incubation, temperature, atmosphere
  • optimal conditions may be empirically determined.
  • humic acid may require different concentrations within media to produce increased efficiency of plating of bacteria isolated from samples, as compared to media that lacks the humic acids (e.g., the optimal concentration of humic acid may not be the same as the optimal concentration of a salt of humic acid).
  • concentrations of any of the various humic acid forms above 0% may be used.
  • humic acid forms may be used at concentrations above 0% and less than about 5% (e.g., 0.25, 0.50, 1.50, 2.00, 2.50%).
  • humic acid forms may be used at concentrations above 0% and less than about 0.25% (e.g., 0.10, 0.15, 0.20, 0.25%).
  • a concentration of humic acid used in the medium is not 0.1% or is above 0.1%.
  • concentrations of humic acid between about 0-5% or 0.05-2.00%) may be used.
  • a concentration of a salt of humic acid below about 0.25% may be used.
  • a concentration of peat of about 0.5% may be used.
  • Nonlimiting examples of dried bacteria may include ⁇ -proteobacteria, ⁇ - proteobacteria, a-proteobacteria, ⁇ -proteobacteria, bacteroidetes, acinobacteria, or firmicutes,
  • the bacteria capable of being recovered using the methods disclosed herein include Gram-negative bacteria.
  • Nonlimiting examples of bacteria recoverable using the methods disclosed herein may be from the genera Acetobacter, Acinetobacter, Aeromonas, Agrobacterium, Alcaligenes, Arcobacter, Bifidobacterium, Bradyrhizobium, Burkholderia, Campylobacter, Citrobacter, Cytophaga, Enterobacter , Enter ococcus, Erwinia, Escherichia, Francisella, Helicobacter, Klebsiella, Lactobacillus, Legionella, Listeria, Oenococcus, Paracoccus, Pasteurella,
  • Nonlimiting examples of bacteria recoverable using the methods disclosed herein may be Acetobacter aceti, Acinetobacter calcoaceticus, Aeromonas hydrophilia, Aeromonas salmonicida, Agrobacterium tumifaciens, Alcaligenes eutrophus, Arcobacter butzleri,
  • Bifidobacterium lactis Bifidobacterium longum, Bifidobacteriumanimalis, Bradyrhizobium japonicum, Bradyrhizobium elkaii, Burkholderia cepacia, Burkholderia pseudomallei,
  • Campylobacter coli Campylobacter jejuni, Campylobacter lari, Citrobacter freundii, Cytophaga allerginae, Enterobacter aerogenes, Enterobacter cloacae, Enterobacter agglomerans,
  • Enterococcus faecalis Enterococcus hirae
  • Enterococcus faecium Enterococcus faecium
  • Erwinia amylovora Enterococcus faecalis, Enterococcus hirae, Enterococcus faecium, Erwinia amylovora,
  • Escherichia coli Francisella tularensis, Helicobacter pylori, Klebsiella aerogenes, Klebsiella pneumoniae, Klebsiella planticola, Lactobacillus plantarum, Lactobacillus lindneri,
  • Lactobacillus paracollinoides Lactobacillus lactus, Legionella pneumophila, Listeria monocyhtogenes, Oenococcus oeni, Paracoccus pantotrophus, Pasteurella piscicida,
  • the bacteria cultured using the methods disclosed herein may not be from the order Actinomycetales (e.g., microbes from this order may be excluded). In some examples, the bacteria cultured using the methods disclosed herein may not be from the phyla Acidobacteria and Verrucomicrobia (e.g., microbes from one or both of these phyla may be excluded). In some examples, the excluded Acidobacteria may belong to subdivision 1 only. In some examples, the excluded Verrucomicrobia may belong to subdivision 4 only.
  • the dried bacteria that have been recovered are enumerated.
  • the enumerated bacteria may be counted directly.
  • the enumerated bacteria may be calculated.
  • a sample or dilution thereof may be cultured on agar- containing nutrient media.
  • One of the media may contain humic acid, a salt thereof, an analog thereof, or peat.
  • the other of the media may not contain humic acid or related substances.
  • the bacteria enumerated on that medium are generally greater than the number of bacteria enumerated on the same medium not containing humic acid. Subtraction of the latter from the former (number on humic acid medium minus number on medium not containing humic acid) yields an estimate of the number of unculturable bacteria in the sample.
  • the bacteria enumerated or counted using medium containing humic acid, salts thereof, analogs thereof, or peat are greater than the number of bacteria enumerated using the same medium without humic acid and/or related substances.
  • the number of bacteria in presence of humic acid and/or related substances may be at least about 1.1-, 1.2-, 1.3-, 1.4-, 1.5-, 1.6-, 1.7-, 1.8-, 1.9-, 2.0-, 2.1-, 2.2-, 2.3-, 2.4-, 2.5-, 2.6-,
  • a method comprising, consisting essentially of, or consisting of:
  • [00170] 6 The method of any one of embodiments 1-5, where the dried sample is spray dried, freeze dried, air dried, or drum dried.
  • non-aqueous carrier includes a monosaccharide, a disaccharide, sugar alcohol, or oligosaccharide.
  • a method for determining viable bacteria in a sample comprising, consisting essentially of, or consisting of:
  • a number of bacterial colonies formed on the medium containing humic acid, a salt thereof, an analog thereof, or peat is at least about 1.1-, 1.2-, 1.3-, 1.4-, 1.5-, 1.6-,
  • [00196] 27 The method of embodiment 26, where the spray-dried sample is stored for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 21, 28, 30, 42, or 64 days, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 16, 20, 24, 48, or 52 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, or 48 months, or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years prior to the plating step.
  • a method for determining viable bacteria comprising, consisting essentially of, or consisting of:
  • a number of bacterial colonies formed on the medium containing humic acid, a salt thereof, an analog thereof, or peat is at least is at least about 1.1-, 1.2-, 1.3-, 1.4-, 1.5- , 1.6-, 1.7-, 1.8-, 1.9-, 2.0-, 2.1-, 2.2-, 2.3-, 2.4-, 2.5-, 2.6-, 2.7-, 2.8-, 2.9- 3.0-, 3.1-, 3.2-, 3.3-, 3.4-, 3.5-, 3.6-, 3.7-, 3.8-, 3.9-, 4.0-, 4.1-, 4.2-, 4.3-, 4.4-, 4.5-, 4.6-, 4.7-, 4.8-, 4.9-, 5.0-, 5.2-, 5.4-, 5.6-, 5.8-, 6.0-, 6.2-, 6.4-, 6.6-, 6.8-, 7.0-, 7.2-, 7.4-, 7.6-, 7.8-,
  • Bradyrhizobium bacteria were formulated, as described below, and then spray dried.
  • Spray drying used a triple nozzle, where the primary formulation, containing the bacteria, was fed through an inner nozzle feed to create the particle core, and a secondary formulation was fed through an outer nozzle, to coat the bacterial core particle.
  • the bacteria used as the starting material was Bradyrhizobium japonicum strain 273 that had been propagated in liquid medium and then had been stored in a liquid formulation in a polypropylene bladder. A volume of material from the bladder was centrifuged and the bacterial pellet was re-suspended in one-tenth of the original volume to obtain a ten-fold concentrated culture.
  • Various formulations containing the concentrated culture were then made with a 70:30 ratio of Maltodextrin Ml 50 to a simple sugar, where the simple sugar for each primary formulation is described in Table 1 below. Some of the formulations contained other additives, as indicated in Table 1.
  • the concentrated culture was added to each formulation at a rate of 69% concentrated culture per weight of formulation material.
  • the final primary formulations consisted by weight of 69% concentrated culture, 21% MALTRIN® Ml 50 (maltodextrin), 9% maltose monohydrate or sorbitol or fructose or xylitol, dependent on formulation, as noted in Table 1 below, 0.08% potassium phosphate dibasic, and 0.02% potassium phosphate monobasic.
  • the final secondary feed formulation consisted of (by weight): 85% deionized water, 0.08% potassium phosphate dibasic, 0.02% potassium phosphate monobasic, 12.75% MALTRIN® Ml 50 (maltodextrin), and 2.25%) maltose monohydrate. Samples were incubated at 25 °C for either 1 or 24 hours as indicated in Table 1 below. All samples were spray dried using a triple nozzle with a target outlet temperature of less than 50°C. The primary feed rate used was 2mL/minute and the secondary feed rate was 11% using a BUCHI Mini Spray Dryer B-290.
  • Spray-dried samples were stored in each of two conditions. One condition was 54% relative humidity at room temperature. The second condition was 65% relative humidity at 25 °C. Stability testing was performed after various days in storage, as noted in Table 2 below. For each time point, the spray-dried samples were re-suspended in standard phosphate buffer solution and serially diluted in 96-well plates to obtain a final dilution range of 10 "2 -10 "7 .
  • Colonies were manually counted and colony forming units (CFU)/mL were calculated. Results for each stability test are found in Tables 2 and 3 below. The recovery results for each formulation stored at 54% relative humidity and room temperature are found in Table 2 below while the results for formulations stored at 65% relative humidity and 25 °C are found in Table 3 below.
  • Table 2 Comparison of Bradyrhizobium japonicum recovery under 54% relative humidity and room temperature conditions in various formulations using YEM and YEMHA agar
  • Values in parenthesis represent normalized data where each CFU/mL count on YEMHA agar was compared to the corresponding CFU/mL count on YEM agar.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Toxicology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne des procédés de récupération de certaines bactéries non cultivables sous une forme cultivable. Les bactéries qui ont été séchées, déshydratées ou desséchées, dans un exemple utilisant un séchage par pulvérisation, sont cultivées plus efficacement au moyen de milieux qui contiennent de l'acide humique, des sels de celui-ci, des analogues de celui-ci, ou de la tourbe, que lors de l'utilisation de milieux qui ne contiennent pas d'acide humique ou de substances apparentées. Lorsque de l'acide humique est présent dans les milieux utilisés dans des dosages d'unités formant des colonies (UFC) de bactéries séchées, des titres plus élevés de bactéries viables sont obtenus par rapport au cas dans lequel le même milieu qui ne contient pas d'acide humique ou de substances apparentées est utilisé.
PCT/US2018/024836 2017-03-30 2018-03-28 Récupération de bactéries séchées WO2018183491A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201762478669P 2017-03-30 2017-03-30
US62/478,669 2017-03-30

Publications (1)

Publication Number Publication Date
WO2018183491A1 true WO2018183491A1 (fr) 2018-10-04

Family

ID=62090036

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2018/024836 WO2018183491A1 (fr) 2017-03-30 2018-03-28 Récupération de bactéries séchées

Country Status (1)

Country Link
WO (1) WO2018183491A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019217255A1 (fr) 2018-05-07 2019-11-14 Novozymes Bioag A/S Isolats de microbacterium et leurs utilisations
WO2020263734A1 (fr) 2019-06-24 2020-12-30 Novozymes Bioag A/S Isolats d'erwinia et leurs utilisations
WO2021086695A1 (fr) 2019-10-29 2021-05-06 Novozymes Bioag A/S Isolats de microbacterium et leurs utilisations
WO2021101949A1 (fr) 2019-11-22 2021-05-27 Novozymes Bioag A/S Isolats de paenibacillus et leurs utilisations
WO2021101937A1 (fr) 2019-11-20 2021-05-27 Novozymes Bioag A/S Isolats de pseudomonas et leurs utilisations
WO2023288294A1 (fr) 2021-07-16 2023-01-19 Novozymes A/S Compositions et procédés pour améliorer la résistance à la pluie de protéines sur des surfaces de plantes
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5854032A (en) 1995-06-07 1998-12-29 Arctech, Inc. Biological production of humic acid and clean fuels from coal
WO2005095579A1 (fr) * 2004-03-31 2005-10-13 Danisco A/S Procede
US20130157848A1 (en) * 2011-12-16 2013-06-20 Novozymes Biologicals Holdings A/S Bradyrhizobium strains
WO2017044473A1 (fr) * 2015-09-11 2017-03-16 Novozymes Bioag A/S Compositions d'inoculant stables et procédés de production de ces compositions

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5854032A (en) 1995-06-07 1998-12-29 Arctech, Inc. Biological production of humic acid and clean fuels from coal
WO2005095579A1 (fr) * 2004-03-31 2005-10-13 Danisco A/S Procede
US20130157848A1 (en) * 2011-12-16 2013-06-20 Novozymes Biologicals Holdings A/S Bradyrhizobium strains
WO2017044473A1 (fr) * 2015-09-11 2017-03-16 Novozymes Bioag A/S Compositions d'inoculant stables et procédés de production de ces compositions

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
BAIRD; ZUBLENA: "SOIL FACTS: USING WETTING AGENTS (NONIONIC SURFACTANTS) ON SOIL", 1993, NORTH CAROLINA COOPERATIVE EXTENSION SERVICE PUBLICATION
BRYAN A. GARNER: "A Dictionary of Modern Legal Usage", vol. 624, 1995
BURGES: "FORMULATION OF MICROBIAL BIOPESTICIDES: BENEFICIAL MICROORGANISMS, NEMATODES AND SEED TREATMENTS", 2012, SPRINGER SCIENCE & BUSINESS MEDIA
HAYAKAWA M ET AL: "Humic acid-vitamin agar, a new medium for the selective isolation of soil actinomycetes", JOURNAL OF FERMENTATION TECHNOLOGY, ELSEVIER, JP, vol. 65, no. 5, 1 January 1987 (1987-01-01), pages 501 - 509, XP025705951, ISSN: 0385-6380, [retrieved on 19870101], DOI: 10.1016/0385-6380(87)90108-7 *
MCCARTY: "WETTING AGENTS", 2001, CLEMSON UNIVERSITY COOPERATIVE EXTENSION SERVICE PUBLICATION
PINTO, D. ET AL.: "Thirty Years of Viable but Nonculturable State Research", CRITICAL REVIEWS IN MICROBIOLOGY, vol. 41, 2015, pages 61 - 76, XP055481183, DOI: doi:10.3109/1040841X.2013.794127
POUCI ET AL., AM. J. AGRIC. BIOL. SCI., vol. 3, no. 1, 2008, pages 299
V. A. LITVIN; R. L. GALAGAN: "Synthesis and Properties of Synthetic Analogs of Natural Humic Acids", RUSSIAN JOURNAL OF APPLIED CHEMISTRY, vol. 85, no. 2, 2012, XP035030845, DOI: doi:10.1134/S1070427212020243
VISSER ET AL: "Physiological action of humic substances on microbial cells", SOIL BIOLOGY AND BIOCHEMISTRY, PERGAMON, OXFORD, GB, vol. 17, no. 4, 1 January 1985 (1985-01-01), pages 457 - 462, XP023671542, ISSN: 0038-0717, [retrieved on 19850101], DOI: 10.1016/0038-0717(85)90009-4 *
XIN-QING ZHAO ET AL: "Screening and identification of actinobacteria from marine sediments: Investigation of potential producers for antimicrobial agents and type I polyketides", WORLD JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, KLUWER ACADEMIC PUBLISHERS, DO, vol. 25, no. 5, 23 January 2009 (2009-01-23), pages 859 - 866, XP019690989, ISSN: 1573-0972 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019217255A1 (fr) 2018-05-07 2019-11-14 Novozymes Bioag A/S Isolats de microbacterium et leurs utilisations
WO2020263734A1 (fr) 2019-06-24 2020-12-30 Novozymes Bioag A/S Isolats d'erwinia et leurs utilisations
WO2021086695A1 (fr) 2019-10-29 2021-05-06 Novozymes Bioag A/S Isolats de microbacterium et leurs utilisations
WO2021101937A1 (fr) 2019-11-20 2021-05-27 Novozymes Bioag A/S Isolats de pseudomonas et leurs utilisations
WO2021101949A1 (fr) 2019-11-22 2021-05-27 Novozymes Bioag A/S Isolats de paenibacillus et leurs utilisations
WO2023288294A1 (fr) 2021-07-16 2023-01-19 Novozymes A/S Compositions et procédés pour améliorer la résistance à la pluie de protéines sur des surfaces de plantes
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes

Similar Documents

Publication Publication Date Title
WO2018183491A1 (fr) Récupération de bactéries séchées
Orozco-Mosqueda et al. The production of ACC deaminase and trehalose by the plant growth promoting bacterium Pseudomonas sp. UW4 synergistically protect tomato plants against salt stress
Bhattacharyya et al. Genome-guided insights into the plant growth promotion capabilities of the physiologically versatile Bacillus aryabhattai strain AB211
Sun et al. Nutrient supplementation of pinewood biochar for use as a bacterial inoculum carrier
Lopez et al. Endophytic bacteria of Mammillaria fraileana, an endemic rock-colonizing cactus of the southern Sonoran Desert
Bale et al. The survival of bacteria exposed to desiccation on surfaces associated with farm buildings
Vassilev et al. Rock phosphate solubilization by immobilized cells of Enterobacter sp. in fermentation and soil conditions
WO2018175681A1 (fr) Récupération de bactéries stressées
Mary et al. Rates of drying and survival of Rhizobium meliloti strains during storage at different relative humidities
Johnson et al. Elicitation of capsaicin production in freely suspended cells and immobilized cell cultures of Capsicum frutescens Mill
Rojas-Tapias et al. Evaluation of three methods for preservation of Azotobacter chroococcum and Azotobacter vinelandii
JP7384347B2 (ja) 微生物凍結乾燥組成物
JP2011223990A (ja) 凍結乾燥菌体の製造方法
US20220053768A1 (en) Methylobacterium compositions for improving corn yield
Park et al. Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors
Patil et al. Azotobacter
Mary et al. Differences among Rhizobium meliloti and Bradyrhizobium japonicum strains in tolerance to desiccation and storage at different relative humidities
Kong et al. Biochemical responses of the mycorrhizae in Pinus massoniana to combined effects of Al, Ca and low pH
Brahmaprakash et al. Role of additives in improving efficiency of bioformulation for plant growth and development
Berninger et al. Zeolite‐based, dry formulations for conservation and practical application of Paraburkholderia phytofirmans PsJN
Salmeron et al. Effects of alachlor and metolachlor on the biological activity of Azospirillum brasilense grown in chemically defined and dialyzed‐soil media
Gracheva et al. Mechanisms of damaging bacteria during lyophilization and protective activity of shielding media
Silva et al. Impact of preservation conditions on fatty acids, xanthan gum production and other characteristics of Xanthomonas campestris pv. mangiferaeindicae IBSBF 2103
Ashraf et al. Bacterial exo-polysaccharides and productivity of the salt affected soils: I. Diversity of exo-polysaccharide producing bacteria from the rhizosphere of wheat (Triticum aestivum L.) grown in normal and saline Pakistani soils
Annizah et al. Phosphate solubilizing bacteria isolated from Tuban mangrove soil, Indonesia

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18721191

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18721191

Country of ref document: EP

Kind code of ref document: A1