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CN-121974501-A - Method for producing water body environment protective agent by fermenting hermetia illucens larva slurry

CN121974501ACN 121974501 ACN121974501 ACN 121974501ACN-121974501-A

Abstract

The invention discloses a method for producing a water body environment protective agent by fermenting hermetia illucens larva slurry, belonging to the technical field of microbial fermentation. The black soldier fly larvae are taken as main raw materials, ground into slurry through a colloid mill, a small amount of carbon source is added, and mixed bacteria of rhodotorula (Phaffia rhodozyma) CGMCC No.29217, bacillus coagulans (Bacillus coagulans) CGMCC No.5233 and clostridium butyricum (Clostridium butyricum) CGMCC No.4729 are utilized for collaborative fermentation, so that the produced product can be widely applied to application scenes such as aquaculture. The fermented black soldier fly larva slurry is rich in astaxanthin, the fermented product has obvious acid fragrance and no peculiar smell, the total number of probiotics is more than or equal to 5.0X10 8 cfu/g, the pH is less than 4.50, the content of micromolecular active polypeptide (acid soluble protein) of the fermented product is improved by 2.5-3 times compared with that before fermentation, the content of antibacterial active substances of the fermented black soldier fly larva slurry detected by using bacillus subtilis CGMCC (B) 63501 as indicator bacteria is more than 1.0g/kg, pathogenic bacteria such as salmonella, vibrio parahaemolyticus, vibrio cholerae and the like are not detected, and normal-temperature storage can be realized. The preparation method has low production cost, simple operation, safety and no residue, and can improve the economic benefit of aquaculture.

Inventors

  • ZHANG SHANTING
  • LIU YAJUN
  • WAN HONGJING
  • WANG YU
  • WANG HAIKUAN

Assignees

  • 惠农达生物技术(江苏)有限公司

Dates

Publication Date
20260505
Application Date
20251231

Claims (6)

  1. 1. A method for producing a water body environment protective agent by fermenting hermetia illucens larva slurry is characterized in that hermetia illucens larva is taken as a main raw material, ground into slurry by a colloid mill, a small amount of carbon source is added, and mixed bacteria of Phaffia rhodozyma, bacillus coagulans and clostridium butyricum are utilized for synergistic fermentation. The rhodotorula is characterized in that the rhodotorula is named as TK1201, is classified as Phaffia rhodozyma, is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms for 12 months 4 days in 2023, and has a preservation address of North Star Xiyu No. 1, 3 in the Chaoyang area of Beijing, and a preservation number of CGMCC No.29217. The bacillus coagulans is named as TQ33, the classification name Bacillus coagulans is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms on 9 th month 9 of 2011, the preservation address is North Star Xiyu No. 1, 3 of the Beijing area, and the preservation number is CGMCC No. 5233. The clostridium butyricum is named as TK2, the classification name Clostridium butyricum is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms on 4 th month and 2 nd day of 2011, the preservation address is North Star Xiyu No. 1 and No. 3 in the Korean region of Beijing city, and the preservation number is CGMCC No. 4729.
  2. 2. The fermented black soldier fly larva slurry according to claim 1 comprises 60% -90% of the black soldier fly larva slurry, 5% -15% of carbon source (glucose, sucrose, brown sugar or molasses, etc.), and 5% -15% of the composite strain.
  3. 3. The water environmental protection agent prepared by fermenting the black soldier fly larva slurry according to claim 1, wherein the fermentation mode is that the black soldier fly larva (fresh or frozen) is ground and crushed into slurry by a colloid mill, the slurry is directly put into a fermentation tank, and simultaneously a carbon source (glucose, sucrose, brown sugar or molasses and the like) and a mixed strain are slowly added into the fermentation tank according to a proportion, the fermentation is carried out for 2-10 days at the temperature of 25-30 ℃ without ventilation, the liquid loading amount is 60% -90%, and the stirring speed is 60-300 rpm. The fermentation process utilizes the residual air in the consumption tank of the rhodozyma and the bacillus coagulans to provide optimal fermentation conditions for anaerobic fermentation of clostridium butyricum, and simultaneously the rhodozyma and the bacillus coagulans can also perform anaerobic fermentation.
  4. 4. The water environmental protection agent prepared by fermenting black soldier fly larva slurry according to claim 1 or 3, wherein the inoculation amount of strain fermentation is 3-6% (v/w) of rhodotorula CGMCC No.29217, 1-5% (v/w) of bacillus coagulans CGMCC No.5233 and 0-5% (v/w) of clostridium butyricum CGMCC No. 4729.
  5. 5. The water environmental protection agent prepared by fermenting the black soldier fly larva slurry according to claim 1 or 3, wherein the fermented black soldier fly larva slurry is rich in astaxanthin, has a sensory color of yellow brown to tan, has a smell of acid and fragrance as a main component, has no peculiar smell, has a pH of less than 4.50, has a probiotics content of more than or equal to 5.0X10 8 CFU/g, does not detect pathogenic bacteria such as salmonella, vibrio parahaemolyticus, vibrio cholerae and the like, and can be stored at normal temperature. The content of the micromolecular active polypeptide (acid soluble protein) of the fermentation product is improved by 2.5-3 times compared with the content of the antibacterial active substance after fermentation of the hermetia illucens larva slurry detected by using bacillus subtilis CGMCC (B) 63501 as indicator bacteria is more than 1.0g/kg.
  6. 6. The water environmental protection agent prepared by fermenting hermetia illucens larva slurry according to claim 1 is suitable for application in aquaculture and other application scenes.

Description

Method for producing water body environment protective agent by fermenting hermetia illucens larva slurry Technical Field The invention belongs to the technical field of microbial fermentation, and relates to an application technology for fermenting black soldier fly larvae serving as a main raw material by grinding the black soldier fly larvae into slurry through a colloid mill and supplementing a small amount of carbon source, wherein the prepared product can be widely applied to aspects of aquaculture and the like by using rhodotorula rubra (Phaffia rhodozyma) CGMCC No.29217, bacillus coagulans (Bacillus coagulans) CGMCC No.5233 and clostridium butyricum (Clostridium butyricum) CGMCC No. 4729. Background Astaxanthin is a lutein carotenoid which is red in color, has strong antioxidant capacity due to a special structure, and is widely applied to industries such as aquaculture, food and the like. At present, astaxanthin sources are mainly produced by chemical synthesis methods, animal and plant extraction methods and microbial fermentation methods. The production process of the chemically synthesized astaxanthin is complex, the pollution is serious, three stereoisomers in the chemically synthesized astaxanthin exist simultaneously, and the utilization rate of animals is low. Compared with astaxanthin produced by chemical synthesis, the astaxanthin produced by an animal and plant extraction method and a microbial fermentation method has the characteristics of high bioavailability, strong coloring capability, high bioactivity and the like, and the development and the utilization of the astaxanthin are attracting more and more attention. Rhodotorula can synthesize astaxanthin in cells, and is one of the most promising and economical natural sources of astaxanthin. The black soldier fly is a saprophytic insect of the genus hermetia of the family hermetiaceae, and the larvae thereof are rich in amino acids, proteins, minerals and other nutritional ingredients, and are rich in antimicrobial peptides, chitin, chitosan and other active substances, so that the immunity and disease resistance of the cultured animals can be improved as baits. In aquaculture, a lot of farmers use the black soldier fly larvae as animal protein feed, and in the process of eating the black soldier fly larvae and shredding the black soldier fly larvae, nutrient substances in the black soldier fly larvae can be lost in a large amount, a large amount of undigested insect skin can be dissected in the alimentary canal of the aquatic animals, and even the insect skin of the black soldier fly larvae can float on the water surface of the mouth of a cultivation pond. The various digestive enzymes, organic acids, vitamins and the like generated by microbial fermentation also greatly improve the nutritive value of the insect protein feed, meanwhile, the microbial fermentation composite protein feed can also obtain antibacterial peptide with physiological activity, improve the biological activity of health-care substances, enable a certain amount of probiotics to exist in fermentation products, be used for improving the intestinal microecological environment of animals, and have great effects on the health and nutrition of the animals. The invention patent No. CN202410134640.5 discloses a method for removing impurities, decontaminating, dehydrating and sterilizing black soldier fly larvae, mixing with an organic carbon source, crushing, fermenting, drying to obtain larva fermented powder, maximally maintaining the nutritive value of the black soldier fly larvae, increasing peptides, functional grease, flavor substances and the like in the fermented powder of the black soldier fly larvae, and reducing the content of harmful bacteria. The patent of the invention with the application number of CN201910436161.8 describes a method for preparing feed protein by fermenting hermetia illucens larvae by utilizing microorganisms, which comprises the steps of pulping the hermetia illucens larvae, mixing the pulp with bean pulp, adding a carbon source, a nitrogen source and a strain for fermentation, and then drying and crushing to obtain the hermetia illucens fermented protein feed. However, the high-temperature heat treatment or the oven-drying pulverization in the above patent results in a loss of a large amount of bioactive substances. The invention patent with the application number of CN202311184508.7 introduces a method for adding concentrated mixed extract of galangal leaves, olive leaves and betel nut pulp into the mixture after scalding and crushing the black soldier fly larvae, and adding the mixture into a compound strain for continuous fermentation, and the mixture is applied to feed for improving the disease resistance of shrimps to aeromonas hydrophila. The invention patent with the application number of CN202110627082.2 discloses a processing method of a fermented feed for aquatic products based on hermetia illucens, which comprises the steps of