Method for improving quality of aquaculture pond water using a nutrient germinant composition and spore incubation method

Patent metadata

Field	Value
Publication number	US-11608283-B2
Application number	US-202016986725-A
Country	US
Kind code	B2
Filing date	Aug 6, 2020
Priority date	May 23, 2014
Publication date	Mar 21, 2023
Grant date	Mar 21, 2023

Abstract

A method for improving the quality of pond water used in aquaculture applications by adding to the pond water active bacteria that are preferably germinated from spores on site using a nutrient-germinant composition and an incubation method for increased spore germination efficiency, in combination with a nitrification enhancement agent such as calcium carbonate or calcified seaweed, and an optional reaction surface area modifier such as calcified seaweed or plastic or metal particles or fragments. The nutrient-germinant composition comprises L-amino acids, D-glucose and/or D-fructose, a phosphate buffer, an industrial preservative, and may include bacteria spores (preferably of one or more Bacillus species) or they may be separately combined for germination. The incubation method comprises heating a nutrient germinant composition and bacteria spores, to a temperature range of 35° C. to 60° C. for around 2 to 60 minutes to produce an incubated bacteria solution that is discharged to the aquaculture application.

First claim

What is claimed is:

1. A method of adding bacteria to water used in an aquaculture application, the method comprising:
heating a portion of a nutrient spore composition to a temperature in a range of around 38° C. to 60° C. at or near a site of the aquaculture application for an incubation period of around 2 minutes to around 6 hours to form a batch of incubated bacteria solution;
periodically repeating the heating step to form additional batches of incubated bacteria solution over the course of a treatment cycle;
dispersing each batch of incubated bacteria solution into the water used in the aquaculture application; and
wherein the nutrient spore composition comprises a nutrient germinant composition in liquid form and bacteria spores.

2. The method of claim 1 wherein the bacteria are one or more species of Bacillus licheniformis and Bacillus subtilis.

3. The method of claim 1 wherein the bacteria is selected from the group consisting of Bacillus amylophilus, Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacteroides ruminocola, Bacteroides ruminocola, Bacterioides suis, Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium thermophilum, Enterococcus cremoris, Enterococcus diacetylactis, Enterococcus faecium, Enterococcus intermedius, Enterococcus lactis, Enterococcus thermophiles, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus cellobiosus, Lactobacillus curvatus, Lactobacillus delbruekii, Lactobacillus farciminis, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus lactis, Lactobacillus plantarum, Lactobacillus reuteri, Leuconostoc mesenteroides, Megasphaera elsdennii, Pediococcus acidilacticii, Pediococcus cerevisiae, Pediococcus pentosaceus, Propionibacterium acidipropionici, Propionibacterium freudenreichii , and Propionibacterium shermanii.

4. The method of claim 1 wherein the nutrient germinant composition comprises:
an L-amino acid;
one or more buffers comprising a phosphate buffer, HEPES, Tris base, or a combination thereof;
an industrial preservative;
optionally D-glucose, or optionally D-fructose, or optionally both D-glucose and D-fructose; and
optionally a source of potassium ions.

5. The method of claim 4 wherein the L-amino acid is L-alanine, L-asparagine, L-valine, L-cysteine, a hydrolysate of soy protein, or a combination thereof and wherein the nutrient germinant composition comprises around 17.8 g/L to 89 g/L total of one or more L-amino acids.

6. The method of claim 1 wherein the nutrient spore composition is a premixed storage stable liquid composition that further comprises a germination inhibitor;
wherein the bacteria spores are spores of a Bacillus species; and
wherein the nutrient germinant composition comprises: (1) around 17.8 g/L to 89 g/L total of one or more L-amino acids comprising L-alanine, L-asparagine, L-valine, L-cysteine, or a hydrolysate of soy protein; and (2) one or more buffers comprising a phosphate buffer, HEPES, Tris base, or a combination thereof.

7. The method of claim 6 wherein the germination inhibitor comprises sodium chloride, D-alanine, or a combination thereof.

8. The method of claim 7 wherein the premixed storage stable liquid composition comprises around 29 g/L to 117 g/L sodium chloride and/or around 8 g/L to 116 g/L D-alanine.

9. The method of claim 5 wherein the one or more buffers comprises around 10-36 g/L of monosodium phosphate and around 30-90 g/L of disodium phosphate.

10. The method of claim 1 wherein the nutrient spore composition is premixed as a concentrated liquid; and
wherein the nutrient germinant composition comprises:
around 8.9-133.5 g/L of one or more L-amino acids;
around 0.8-3.3 g/L total of one or more industrial preservatives;
around 40-126 g/L total of one or more phosphate buffers, around 15-61 g/L Tris base, or around 32.5-97.5 g/L HEPES, or a combination thereof;
optionally around 18-54 g/L of D-glucose, D-fructose, or a combination thereof; and
optionally around 7.4-22.2 g/L of KCl; and
wherein the bacteria spores comprise one or more species of Bacillus.

11. The method of claim 10 further comprising:
adding a diluent to the nutrient spore composition prior to or during heating; and
mixing the diluted nutrient spore composition during the incubation period; and
wherein the diluted nutrient spore composition has a concentration of around 0.1° A to 10% of the concentrated liquid.

12. The method of claim 1 further comprising:
providing the nutrient germinant composition and providing a separate spore composition in liquid form;
mixing the nutrient germinant composition with the spore composition at or near the site of the aquaculture application to form the nutrient spore composition; and
wherein the spore composition comprises:
the bacteria spores comprising one or more Bacillus species in spore form;
about 0.002 to 5.0% by weight thickener;
about 0.01 to 2.0% by weight total of one or more acids or salts of acids;
wherein the spore composition has a pH of around 4.5 to around 5.5; and
wherein the percentages are by weight of the spore composition.

13. The method of claim 4 further comprising:
providing the nutrient germinant composition and providing a separate spore composition in liquid form;
mixing the nutrient germinant composition with the spore composition at or near the site of the aquaculture application to form the nutrient spore composition; and
wherein the spore composition comprises:
the bacteria spores comprising one or more Bacillus species in spore form;
one or more acids or salts of acids;
a thickener; and
wherein the spore composition has a pH of around 4.5 to around 5.5.

14. The method of claim 13 wherein the spore Bacillus species are one or more of: Bacillus pumilus, Bacillus licheniformis, Bacillus amylophilus, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus clausii, Bacillus firmus, Bacillus megaterium, Bacillus mesentericus, Bacillus subtilis var natto, or Bacillus toyonensis.

15. The method of claim 13 wherein the acids or salts of acids are one or more of acetic acid, citric acid, fumaric acid, propionic acid, sodium propionate, calcium propionate, formic acid, sodium formate, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, or calcium sorbate.

16. The method of claim 13 wherein the spore composition comprises:
about 0.002 to 5.0% by weight thickener;
about 0.01 to 1% by weight total of one or more acids or salts of acids; and
about 0.00005 to 3.0% by weight of a surfactant;
wherein the percentages are by weight of the spore composition.

17. The method of claim 3 wherein the incubation period is around 2 minutes to around 5 minutes.

18. The method of claim 17 wherein the aquaculture application is a growing pond containing fish or eel.

19. The method of claim 1 wherein the incubation period is around 4 to 6 hours.

20. The method of claim 19 wherein the aquaculture application is a growing pond containing shrimp.

21. The method of claim 1 wherein the temperature is in a range of around 38° C. to 50° C.

22. The method of claim 1 wherein the temperature is in a range of around 41° C. to 44° C.

23. The method of claim 1 wherein the nutrient germinant composition does not include fructose or glucose.

24. The method of claim 4 w

Assignees

• Nch Corp

Inventors

• Greenwald Charles
• Everett Gabriel F K
• Pruitt Judy
• Rosmarin Amanda
• Church Jordan
• Aberle Daniel
• Aboagye George
• White Skylar Rae
• Cao Haibo
• Zetena Christopher
• Gillespie Kelly A

Classifications

• C02F2101/105

  Phosphorus compounds · CPC title

• A01K63/04

  Arrangements for treating water specially adapted to receptacles for live fish · CPC title

• C02F2103/20

  from animal husbandry · CPC title

• C02F3/302

  Nitrification and denitrification treatment (C02F3/308 takes precedence) · CPC title

• C02F2101/16

  Nitrogen compounds, e.g. ammonia · CPC title