CN105110489A - Water-purifying and weed-protecting biological agent for shrimp and crab culture in high-temperature period as well as preparation method and application of biological agent - Google Patents
Water-purifying and weed-protecting biological agent for shrimp and crab culture in high-temperature period as well as preparation method and application of biological agent Download PDFInfo
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Abstract
The invention relates to a water-purifying and weed-protecting biological agent for shrimp and crab culture in the high-temperature period as well as a preparation method and an application of the biological agent and belongs to the technical field of aquaculture. The method mainly comprises steps as follows: a high-concentration culture of lactobacillus breris and enterococcus faecalis and potassium fulvic acid are taken and fully mixed in the weight ratio, sealing is performed after filling by a filling machine, and the net content of each package is 5 L. The biological agent can be directly added to a culture water body, relieve dirt on aquatic weed surfaces, resist harmful microbes on the aquatic weed surfaces, prevent aquatic weeds from rotting, effectively degrade organic matters such as protein, carbohydrates and the like in the culture water body, remove harmful substances such as ammonia nitrogen, nitrite and the like in the water body, promote intake of shrimps and crabs and optimize and repair the culture environment, cannot increase the oxygen consumption rate of the water body and creates a healthy and good water body ecological environment for the shrimps and crabs in the high-temperature period.
Description
Technical field
The present invention relates to a kind of pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation and its preparation method and application, belong to technical field of aquaculture.
Background technology
As the high protein animal source of high-quality, Shrimp waste is extensively concerned because having the features such as meat exquisiteness, delicious flavour and nutritive value are high.The share of Shrimp waste cultivation shared by the aquaculture of China is also improving year by year.Because Shrimp waste needs certain sheltered situation during degenerate cllipticity in water body, therefore at present in the freshwater shrimps and crabs class pond culture of China, what mainly adopt is kind of an ecomodel for grass cultivation.
Pasture and water as the primary producer in pond ecosystem, for maintain pond ecosystem stable, improve cultured output and benefit has great importance.Cost is low, reusing is high, detergent power is strong and provide the plurality of advantages such as sheltered situation for Shrimp waste and enjoy the attention and concern that cultivate dealer because having for pasture and water.But the health of pasture and water needs meticulously to safeguard in pond.In the pliotherm period, along with the continuous rising of water temperature, the harmful microorganism in aquaculture water increases severely, and fertilizer remaining in water body, feed, biological waste and meta-bolites are constantly accumulated, and are deposited on water bottom and pasture and water.These materials adhere to pasture and water surface and form mud, affect the attachment of shrimp crab and the photosynthetic of pasture and water and respiration.Time one is long will cause pasture and water surface dirt to be festered, withered and dead, affect quality of water environment, even to Shrimp waste generation toxic effect.And the above-mentioned substance decomposable asymmetric choice net being deposited on water bottom produces the objectionable impurities such as ammonia nitrogen, nitrite, reduce water body dissolved oxygen, affect quality of water environment, harm shrimp crab is healthy.Therefore how under the prerequisite ensureing aquaculture organism safety, effectively purifying aquaculture water quality, guarantee pasture and water healthy growth are the focuses that cultivation dealer pays close attention to.
In recent years, relevant investigation and application achieves certain effect, but there are problems equally.In the purification of shrimp crab aquaculture water, adopt microbiological manipulation technology safely and efficiently, as EM bacterium, genus bacillus etc., cost is low, easy to use, Be very effective more, effectively can control the organic contamination of water body, reduce the pollution level of ammonia nitrogen, nitrite in water body.But these microbial preparations mostly are aerobic microbiological, use at high temperature season and can strengthen water body oxygen consumption, can water hypoxia be caused, simultaneously to removing pasture and water surface sludge, preventing pasture and water from rotting and withered poor effect.On pasture and water health maintenance, adopt at present a class chemical substance such as tensio-active agent, flocculation agent more, although there is certain effect in a short time, need often to use, impact that can be certain on the generation of water ecology balance.Therefore the cultivation of shrimp crab is the health control of water quality and pasture and water in major tasks of pliotherm period, need exploitation a kind of eco-friendly, integrate the functional living being preparation purifying water, prevent anoxic, protect the strong grass of grass, for the benign development of shrimp crab cultivation provides technical support.
Summary of the invention
The object of the invention is to overcome deficiency of the prior art, a kind of pliotherm period shrimp crab cultivation water purification is provided to protect biotechnological formulation of grass and its preparation method and application, utilize short lactobacillus, the high-concentration culturing thing of enterococcus faecalis and potassium fulvate, directly aquaculture water is dropped into after rationally processing and be composite, pasture and water surface can be alleviated dirty, the harmful microorganism on antagonism pasture and water surface, prevent pasture and water from rotting, protein in effective degrading cultivation water, the organic substances such as carbohydrate, remove ammonia nitrogen in water body, the objectionable impuritiess such as nitrite, do not increase water body consumption rate, promote ingesting of Shrimp waste, optimize and repair breeding environment, Shrimp waste for the pliotherm period builds a water ecological setting in good health.
According to technical scheme provided by the invention, a kind of pliotherm period shrimp crab cultivation water purification protects careless biotechnological formulation, formula rate is as follows by weight: the high-concentration culturing thing of short lactobacillus 50 ~ 60 parts, the high-concentration culturing thing of enterococcus faecalis 35 ~ 45 parts, potassium fulvate liquid 5 ~ 10 parts; Above-mentioned mixing of materials is evenly loaded in packing bottle afterwards, obtains product pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation.
Described pliotherm period shrimp crab cultivation water purification protects the preparation method of careless biotechnological formulation, and step is by weight:
(1) the high-concentration culturing thing preparation of short lactobacillus: get a ring short lactobacillus CGMCC1.558 lawn from test tube slant, be inoculated in the MRS liquid nutrient medium of 50 ~ 100mL sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 40h; Then accessed 5 ~ 8L in the MRS liquid nutrient medium of sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 35h, made bacterial classification seed liquor for subsequent use; Now viable count>=10 of bacterium liquid
9cFUmL
-1;
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300 ~ 350L, cool for subsequent use after 121 DEG C of sterilizing 15min; By above-mentioned bacterial classification seed liquor access fermentor tank, at 32 ~ 37 DEG C, rotating speed 60 ~ 80rmin
-1condition under cultivate 30 ~ 35h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use, obtain the high-concentration culturing thing of short lactobacillus;
(2) the high-concentration culturing thing preparation of enterococcus faecalis: get a ring enterococcus faecalis CGMCC1.2135 lawn from test tube slant, be inoculated in the MRS liquid nutrient medium of 50 ~ 100mL sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 40h; Then accessed 5 ~ 8L in the MRS liquid nutrient medium of sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 35h, made bacterial classification seed liquor for subsequent use; Now viable count>=10 of bacterium liquid
9cFUmL
-1;
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300 ~ 350L, cool for subsequent use after 121 DEG C of sterilizing 15min; By above-mentioned bacterial classification seed liquor access fermentor tank, at 32 ~ 37 DEG C, rotating speed 60 ~ 80rmin
-1condition under cultivate 30 ~ 35h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use, obtain the high-concentration culturing thing of enterococcus faecalis;
(3) preparation of potassium fulvate liquid: get potassium fulvate 25 ~ 40 parts, adds in 65 ~ 80 parts of water, dissolves in 121 DEG C of sterilizing 15min after stirring, for subsequent use after cooling, obtains potassium fulvate liquid;
(4) mix: the high-concentration culturing thing 50 ~ 60 parts getting the obtained short lactobacillus of step (1), the high-concentration culturing thing 35 ~ 45 parts of the enterococcus faecalis that step (2) is worth, the potassium fulvate liquid 5 ~ 10 parts that step (3) is obtained; Above-mentioned mixing of materials is evenly loaded in packing bottle afterwards, obtains product pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation.
Described MRS liquid nutrient medium is, extractum carnis 10gL
-1, peptone 10gL
-1, yeast extract paste 5gL
-1, glucose 5gL
-1, citric acid diamines 2gL
-1, tween 80 1gL
-1, sodium acetate 5gL
-1, dipotassium hydrogen phosphate 2gL
-1, magnesium sulfate 0.2gL
-1, manganous sulfate 0.05gL
-1, calcium carbonate 20gL
-1, adjust ph 6.2 ~ 6.6, cools for subsequent use after 121 DEG C of sterilizing 15min;
Described liquid fermentation medium, its component ratio by mass number meter: 15 parts, molasses, soyflour 5 parts, fresh fish cream 5 parts, yeast extract paste 5 parts, sodium acetate 2 parts, 0.1 part, magnesium sulfate, dipotassium hydrogen phosphate 1.0 parts, 66.9 parts, water, dissolves mixing, regulator solution pH value 6.2 ~ 6.6 under room temperature in fermentor tank, 121 DEG C of sterilizing 15min, for subsequent use after cooling.
Sugar content >=55% in described molasses.
The reagent being used for regulating pH is lactic acid and sodium bicarbonate.
Described pliotherm period shrimp crab cultivation water purification protects the application of careless biotechnological formulation, and pliotherm period shrimp crab cultivation water purification is protected careless biotechnological formulation and is used in cultivating pool, consumption is 300 ~ 1000g/ mu meter Shui Shen, is used in conjunction 1 ~ 2 time.
In the present invention, the action principle of each component is as follows:
1, short lactobacillus: short lactobacillus (
lactobacillusbrevis) belong to Bacterium lacticum order, lactobacillaceae, short lactobacillus genus, distributed more widely, have on plant stem-leaf surface, more common in pickles, maximum with small intestine in humans and animals body Digestive tract.Short lactobacillus cell is shaft-like, does not form gemma, Gram-positive; Catalase, oxidase negative; Do not grow in atmosphere, amphimicrobian, glucose fermentation, Sunmorl N 60S, pectinose, fructose, ribose, lactose, maltose, melibiose, semi-lactosi produce acid; Can grow in the environment close with human small intestine's gallbladder salinity; The suitable concentration of salt is roughly 2% ~ 3%; Well-grown near pH6.0, also can show good acid resistance in the environment of pH3.0, but pH is too low or too highly it all can be suppressed to grow; The optimal temperature of fermentation is 29.5 ~ 30 DEG C.
Short lactobacillus can produce natural microbiotic-lactobacillin (Lactobacillin) voluntarily, and lactobacillin has stimulates the growth of body secretes antibody, raising immunizing power and promotion probiotics, selectivity kills the critical functions such as pathogenic bacterium.The polysaccharide that short lactobacillus produces can induce body to produce Interferon, rabbit, causes non-specific immunity.Short lactobacillus has stronger removal nitrite ability, if nitrite content is at 250mgL
-1within, inoculation short lactobacillus 48h nitrite just can all be removed, and the optimum pH that short lactobacillus removes nitrite is 5.0 ~ 6.0, and optimum temperuture is 30 DEG C; If nitrite content is at 200mgL
-1within, short lactobacillus has extremely significant linear relationship to the removal amount of nitrite and concentration of substrate.
The present invention utilizes its secretion lactic acid and lactobacillus usually remove pasture and water surface sludge and prevent from rotting, and transforms the nitrite removed in water body simultaneously, the pliotherm period uses not water consumption body dissolved oxygen etc.
2, enterococcus faecalis: enterococcus faecalis (
enterococcusfaecalis) cry streptococcus faecium again, apply C polysaccharide antigen, according to Lan Shi serological classification, suis can be divided into many groups.Streptococcus faecium belongs to D group.D group streptococcus divides faecalis and non-faecalis two class.The former comprises enterococcus faecalis, faecium and streptococcus durans, and the latter has streptococcus bovis and streptococcus equinus.Enterococcus faecalis is additive for farm animal feed, can prevent and treat the diarrhea that cub fledgling poultry causes because of alteration of intestinal flora, has the effect of the imbalance of regulating intestinal canal normal microflora and growth promoting effects.Enterococcus faecalis bacterium shape circle or oval, can extend along the direction of chain, diameter 0.5 ~ 1.0 μm, and great majority become two or short catenation, usually do not move.On rich medium, bacterium colony is large and smooth, diameter 1 ~ 2mm, Quan Yuan, rare pigment.Its nutritional requirement is low, and ordinary nutrient agar also can grow.Can 10 ~ 45 DEG C, pH value 9.6 or containing growing in 6.5%NaCl broth culture, and ability 65 DEG C of 30min.It can utilize arginine for the energy, fermentation sorbyl alcohol, nonfermented pectinose.Simple substratum grows and does not need folic acid.It produces the nutritive substance useful to cultivated animals in metabolism and growth process, as lactic acid, amino acid, VITAMIN, enzyme and antibacterial substance.In feed, add enterococcus faecalis, not only preservative activity is played to feed, and the local flavor of feed can be increased, promote the appetite of cultivated animals.Partial protein can be resolved into acid amide and amino acid by enterococcus faecalis, and the nitrogen-free extract of most carbohydrate is converted into lactic acid, and can make the fiber deliquescing in feed, allows the conversion specific absorption of feed uprise.It also can be used as pond water quality and substrate modifier, organic debris in degrading cultivation environment, reduces the hazardous and noxious substances such as ammonia nitrogen and nitrite, plays Optimal culture environment, promote the effects such as water surrounding benign ecological cycles, be conducive to the digestibility improving feed simultaneously.
The lactic acid that the present invention utilizes it to secrete, amino acid, VITAMIN, enzyme and antibacterial substance are to prevent pasture and water from rotting, to promote pasture and water growth, organic debris in degrading cultivation environment, reduce the hazardous and noxious substances such as ammonia nitrogen and nitrite, the pliotherm period uses not water consumption body dissolved oxygen simultaneously.
3, potassium fulvate: potassium fulvate is that a kind of pure natural mineral substance active potassium element is fertile, include trace element, rare earth element,
plant-growth regulator, the multiple nutritional components such as viral inhibitors, make that nutrient is more sufficient, supply is more reasonable, thus avoid the generation of the various physiological disturbances that crop is caused because lacking element, make that crop plant type is more vigorous, leaf look more dark green, lodging tolerance is stronger.Potassium fulvate can supplement the nutrient run off in soil timely, makes soil activating, has vitality, decreases the continuous cropping disease that in soil, nutrient is caused by taken in excess.Potassium fulvate not pure molecular compound, but a kind of macromolecular structure of inhomogenous plyability and the extremely complicated mixture of composition.Except high-content xanthohumic acid except, also be rich in almost whole amino acid, nitrogen, phosphorus, potassium, multiple enzyme, carbohydrate (oligose, fructose etc.) protein, nucleic acid, humic acid and the nutritions such as VC, VE and a large amount of vitamin B group required in growing process, there is the short long factor of the unknown of high biological activity function; Its complex ability is strong, can improve absorption and the running of micro elements by plants; Deflocculate, tool cushion, and solubility property is good, strong with metal ion interaction ability; Be conducive to the absorption of plant to moisture content and nutritive element, and then improve crop yield, improve crop quality.
The present invention utilizes it to the premium properties of plant to safeguard the health of pasture and water, removes the impact of metal in water body.
Biological material specimens preservation:
Short lactobacillus
lactobacillusbrevis, CGMCC1.558, is disclosed in China General Microbiological culture presevation administrative center, network address
http:// www.bnbio.com/p_55/p_50855.html; Enterococcus faecalis
enterococcusfaecalis, CGMCC1.2135, is disclosed in China General Microbiological culture presevation administrative center, network address
http:// www.bnbio.com/p_31/p_52431.html.
Beneficial effect of the present invention:
The present invention utilizes short lactobacillus, the high-concentration culturing thing of enterococcus faecalis and potassium fulvate, directly aquaculture water is dropped into after rationally processing and be composite, can alleviate that pasture and water surface is dirty, the harmful microorganism on antagonism pasture and water surface, prevent pasture and water from rotting, the organic substances such as the protein in effective degrading cultivation water, carbohydrate, remove the objectionable impurities such as ammonia nitrogen, nitrite in water body, do not increase water body consumption rate, promote ingesting of Shrimp waste, optimize and repair breeding environment, the Shrimp waste for the pliotherm period builds a water ecological setting in good health.
It is easy to use that pliotherm period shrimp crab cultivation water purification of the present invention protects careless biotechnological formulation, and usage quantity is little, and the consumption of every mu of water surface every meter of depth of water is only 300 ~ 1000mL; Can not water hypoxia be caused after using, significantly can reduce the content of the objectionable impuritiess such as organic substance in water body, ammonia nitrogen, nitrite, hydrogen sulfide, improve the environmental quality of water quality and substrate; Can pathogenic micro-organism in antagonism water body, reduce pasture and water surface dirty, prevent pasture and water from rotting, promote pasture and water growth, improve situation of ingesting and the life condition of pliotherm period shrimp crab, improve cultured output and quality.
Embodiment
Following fermentor tank, mixing machine, wrapping machine etc. are conventional equipment, commercially buy, without any particular requirement, adopt common working method.Material convenient sources of the present invention, all can commercially buy.
The present invention below will be further described in conjunction with the embodiments:
Embodiment 1
A kind of pliotherm period shrimp crab cultivation water purification of the present invention protects careless biotechnological formulation and preparation method, adopts following processing step:
(1) short lactobacillus high-concentration culturing thing preparation: get from test tube slant a ring short lactobacillus (
lactobacillusbrevis, CGMCC1.558, by China General Microbiological culture presevation, administrative center provides) lawn, be inoculated in the MRS liquid nutrient medium of 50mL sterilizing, in 35 DEG C of quiescent culture 36h.Then accessed 5L in the MRS liquid nutrient medium of sterilizing, in 35 DEG C of quiescent culture 30h, made bacterial classification seed liquor for subsequent use.Now viable count>=10 of bacterium liquid
9cFUmL
-1.
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 350L, cool for subsequent use after 121 DEG C of sterilizing 15min.By above-mentioned bacterial classification seed liquor access fermentor tank, at 35 DEG C, rotating speed 60rmin
-1condition under cultivate 30h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use.
(2) enterococcus faecalis high-concentration culturing thing preparation: get from test tube slant a ring enterococcus faecalis (
enterococcusfaecalis, CGMCC1.2135, by China General Microbiological culture presevation, administrative center provides) lawn, be inoculated in the MRS liquid nutrient medium of 50mL sterilizing, in 35 DEG C of quiescent culture 36h.Then accessed 5L in the MRS liquid nutrient medium of sterilizing, in 35 DEG C of quiescent culture 30h, made bacterial classification seed liquor for subsequent use.Now viable count>=10 of bacterium liquid
9cFUmL
-1.
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 350L, cool for subsequent use after 121 DEG C of sterilizing 15min.By above-mentioned bacterial classification seed liquor access fermentor tank, at 35 DEG C, rotating speed 60rmin
-1condition under cultivate 30h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use.
Described MRS liquid nutrient medium is, extractum carnis 10gL
-1, peptone 10gL
-1, yeast extract paste 5gL
-1, glucose 5gL
-1, citric acid diamines 2gL
-1, tween 80 1gL
-1, sodium acetate 5gL
-1, dipotassium hydrogen phosphate 2gL
-1, magnesium sulfate 0.2gL
-1, manganous sulfate 0.05gL
-1, calcium carbonate 20gL
-1, regulate about pH6.5, cool for subsequent use after 121 DEG C of sterilizing 15min.
Described liquid fermentation medium, its component ratio by mass number meter: 15 parts, molasses (sugar content >=55%), soyflour 5 parts, fresh fish cream 5 parts, yeast extract paste 5 parts, sodium acetate 2 parts, 0.1 part, magnesium sulfate, dipotassium hydrogen phosphate 1.0 parts, 66.9 parts, water, dissolves mixing, regulator solution pH value to 6.5 under room temperature in fermentor tank, 121 DEG C of sterilizing 15min, for subsequent use after cooling.
(3) preparation of potassium fulvate liquid: by weight, gets potassium fulvate 30 parts, adds in 70 parts of water, dissolves in 121 DEG C of sterilizing 15min after stirring, for subsequent use after cooling.
(4) mix: count by weight, get the high-concentration culturing thing 60 parts of short lactobacillus, the high-concentration culturing thing of enterococcus faecalis 35 parts, potassium fulvate liquid 5 parts, mix in rear loading packing bottle, obtain product pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation.
Embodiment 2
A kind of pliotherm period shrimp crab cultivation water purification of the present invention protects careless biotechnological formulation and preparation method, adopts following processing step:
(1) short lactobacillus high-concentration culturing thing preparation: get from test tube slant a ring short lactobacillus (
lactobacillusbrevis, CGMCC1.558, by China General Microbiological culture presevation, administrative center provides) lawn, be inoculated in the MRS liquid nutrient medium of 75mL sterilizing, in 32 DEG C of quiescent culture 40h.Then accessed 6L in the MRS liquid nutrient medium of sterilizing, in 32 DEG C of quiescent culture 35h, made bacterial classification seed liquor for subsequent use.Now viable count>=10 of bacterium liquid
9cFUmL
-1.
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300L, cool for subsequent use after 121 DEG C of sterilizing 15min.By above-mentioned bacterial classification seed liquor access fermentor tank, at 32 DEG C, rotating speed 70rmin
-1condition under cultivate 34h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use.
(2) enterococcus faecalis high-concentration culturing thing preparation: get from test tube slant a ring enterococcus faecalis (
enterococcusfaecalis, CGMCC1.2135, by China General Microbiological culture presevation, administrative center provides) lawn, be inoculated in the MRS liquid nutrient medium of 75mL sterilizing, in 32 DEG C of quiescent culture 40h.Then accessed 6L in the MRS liquid nutrient medium of sterilizing, in 32 DEG C of quiescent culture 35h, made bacterial classification seed liquor for subsequent use.Now viable count>=10 of bacterium liquid
9cFUmL
-1.
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300L, cool for subsequent use after 121 DEG C of sterilizing 15min.By above-mentioned bacterial classification seed liquor access fermentor tank, at 32 DEG C, rotating speed 70rmin
-1condition under cultivate 35h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use.
Described MRS liquid nutrient medium is, extractum carnis 10gL
-1, peptone 10gL
-1, yeast extract paste 5gL
-1, glucose 5gL
-1, citric acid diamines 2gL
-1, tween 80 1gL
-1, sodium acetate 5gL
-1, dipotassium hydrogen phosphate 2gL
-1, magnesium sulfate 0.2gL
-1, manganous sulfate 0.05gL
-1, calcium carbonate 20gL
-1, regulate about pH6.2, cool for subsequent use after 121 DEG C of sterilizing 15min.
Described liquid fermentation medium, its component ratio by mass number meter: 15 parts, molasses (sugar content >=55%), soyflour 5 parts, fresh fish cream 5 parts, yeast extract paste 5 parts, sodium acetate 2 parts, 0.1 part, magnesium sulfate, dipotassium hydrogen phosphate 1.0 parts, 66.9 parts, water, dissolves mixing, regulator solution pH value to 6.2 under room temperature in fermentor tank, 121 DEG C of sterilizing 15min, for subsequent use after cooling.
(3) preparation of potassium fulvate liquid: by weight, gets potassium fulvate 35 parts, adds in 65 parts of water, dissolves in 121 DEG C of sterilizing 15min after stirring, for subsequent use after cooling.
(4) mix: count by weight, get the high-concentration culturing thing 55 parts of short lactobacillus, the high-concentration culturing thing of enterococcus faecalis 40 parts, potassium fulvate liquid 5 parts, mix in rear loading packing bottle, obtain product pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation.
Embodiment 3:
A kind of pliotherm period shrimp crab cultivation water purification of the present invention protects careless biotechnological formulation and preparation method, adopts following processing step:
(1) short lactobacillus high-concentration culturing thing preparation: get from test tube slant a ring short lactobacillus (
lactobacillusbrevis, CGMCC1.558, by China General Microbiological culture presevation, administrative center provides) lawn, be inoculated in the MRS liquid nutrient medium of 100mL sterilizing, in 37 DEG C of quiescent culture 35h.Then accessed 8L in the MRS liquid nutrient medium of sterilizing, in 37 DEG C of quiescent culture 32h, made bacterial classification seed liquor for subsequent use.Now viable count>=10 of bacterium liquid
9cFUmL
-1.
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300L, cool for subsequent use after 121 DEG C of sterilizing 15min.By above-mentioned bacterial classification seed liquor access fermentor tank, at 37 DEG C, rotating speed 80rmin
-1condition under cultivate 30h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use.
(2) enterococcus faecalis high-concentration culturing thing preparation: get from test tube slant a ring enterococcus faecalis (
enterococcusfaecalis, CGMCC1.2135, by China General Microbiological culture presevation, administrative center provides) lawn, be inoculated in the MRS liquid nutrient medium of 100mL sterilizing, in 37 DEG C of quiescent culture 35h.Then accessed 5L in the MRS liquid nutrient medium of sterilizing, in 37 DEG C of quiescent culture 32h, made bacterial classification seed liquor for subsequent use.Now viable count>=10 of bacterium liquid
9cFUmL
-1.
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300L, cool for subsequent use after 121 DEG C of sterilizing 15min.By above-mentioned bacterial classification seed liquor access fermentor tank, at 37 DEG C, rotating speed 60rmin
-1condition under cultivate 30h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use.
Described MRS liquid nutrient medium is, extractum carnis 10gL
-1, peptone 10gL
-1, yeast extract paste 5gL
-1, glucose 5gL
-1, citric acid diamines 2gL
-1, tween 80 1gL
-1, sodium acetate 5gL
-1, dipotassium hydrogen phosphate 2gL
-1, magnesium sulfate 0.2gL
-1, manganous sulfate 0.05gL
-1, calcium carbonate 20gL
-1, regulate about pH6.6, cool for subsequent use after 121 DEG C of sterilizing 15min.
Described liquid fermentation medium, its component ratio by mass number meter: 15 parts, molasses (sugar content >=55%), soyflour 5 parts, fresh fish cream 5 parts, yeast extract paste 5 parts, sodium acetate 2 parts, 0.1 part, magnesium sulfate, dipotassium hydrogen phosphate 1.0 parts, 66.9 parts, water, dissolves mixing, regulator solution pH value to 6.5 under room temperature in fermentor tank, 121 DEG C of sterilizing 15min, for subsequent use after cooling.
(3) preparation of potassium fulvate liquid: by weight, gets potassium fulvate 25 parts, adds in 75 parts of water, dissolves in 121 DEG C of sterilizing 15min after stirring, for subsequent use after cooling.
(4) mix: count by weight, get the high-concentration culturing thing 58 parts of short lactobacillus, the high-concentration culturing thing of enterococcus faecalis 35 parts, potassium fulvate liquid 7 parts, mix in rear loading packing bottle, obtain product pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation.
Application Example 1
Product prepared by Example 1 uses at Gaochun, Jiangsu one river crab cultivation pond.
Area 10 mu, pond water quality overrich during summer high temperature (August), water colour is bad, and in deep green, substrate is a bit black smelly, and water body ammonia nitrogen reaches 1.2mgL
-1, nitrite is 0.3mgL
-1, DO is 4.2mgL
-1algae, the mud of pasture and water surface attachment are a lot, and in blackish green, blade is withered and yellow, and surface bacteria is 3 × 10
5individual cm
-1, it is smelly that root is sent out rotten.Water transparency is low, and be 18cm, river crab shell is unsuccessful.One morning 10 the biotechnological formulation of above-mentioned formula is used in cultivating pool, consumption is 1000g/ mu meter Shui Shen.0.18mgL is reduced to the nitrite in latter 2 days ponds
-1, ammonia nitrogen reduces to 0.8mgL
-1, DO is 4.8mgL
-1, water quality becomes salubrious; After 5 days, water transparency increases to 25cm, and substrate stink disappears, and pasture and water surface sludge reduces, and the withered and yellow improvement of pasture and water blade, surface bacteria reduces to 6 × 10
3individual cm
-1, river crab activity recovers normal, and shell increases.
Application Example 2
Product prepared by Example 2 uses at Xinghua, Jiangsu one river crab cultivation pond.
Area 20 mu, pond water quality overrich during summer high temperature (August), water colour is bad, and in deep green, substrate is a bit black smelly, and water body ammonia nitrogen reaches 1.5mgL
-1, nitrite is 0.28mgL
-1, DO is 4.0mgL
-1algae, the mud of pasture and water surface attachment are a lot, and in blackish green, blade is withered and yellow, and surface bacteria is 9 × 10
5individual cm
-1, it is smelly that root is sent out rotten.Water transparency is low, and be 15cm, river crab shell is unsuccessful.One morning 10 the biotechnological formulation of above-mentioned formula is used in cultivating pool, consumption is 500g/ mu meter Shui Shen.0.20mgL is reduced to the nitrite in latter 2 days ponds
-1, ammonia nitrogen reduces to 1.0mgL
-1, DO is 4.5mgL
-1, water quality becomes salubrious; After 5 days, water transparency increases to 23cm, and substrate stink disappears, and pasture and water surface sludge reduces, and the withered and yellow improvement of pasture and water blade, surface bacteria reduces to 8 × 10
2individual cm
-1, river crab activity recovers normal, and shell increases.
Application Example 3
Product prepared by Example 3 uses at changzhou one river prawn cultivating pool.
Area 10 mu, pond water quality overrich during summer high temperature (July), water colour is bad, and in deep green, substrate is a bit black smelly, and water body ammonia nitrogen reaches 1.8mgL
-1, nitrite is 0.35mgL
-1, DO is 3.8mgL
-1algae, the mud of pasture and water surface attachment are a lot, and in blackish green, blade is withered and yellow, and surface bacteria is 6 × 10
5individual cm
-1, it is smelly that root is sent out rotten.Water transparency is low, is 22cm, river prawn poor growth.One morning 10 the biotechnological formulation of above-mentioned formula is used in cultivating pool, consumption is 1000g/ mu meter Shui Shen.0.10mgL is reduced to the nitrite in latter 2 days ponds
-1, ammonia nitrogen reduces to 0.75mgL
-1, DO is 5.0mgL
-1, water quality becomes salubrious; After 5 days, water transparency increases to 30cm, and substrate stink disappears, and pasture and water surface sludge reduces, and the withered and yellow improvement of pasture and water blade, surface bacteria reduces to 2 × 10
3individual cm
-1, river prawn activity recovers normal, increase of ingesting.
Claims (6)
1. pliotherm period shrimp crab cultivation water purification protects a careless biotechnological formulation, it is characterized in that formula rate is as follows by weight: the high-concentration culturing thing of short lactobacillus 50 ~ 60 parts, the high-concentration culturing thing of enterococcus faecalis 35 ~ 45 parts, potassium fulvate liquid 5 ~ 10 parts; Above-mentioned mixing of materials is evenly loaded in packing bottle afterwards, obtains product pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation.
2. described in claim 1, pliotherm period shrimp crab cultivation water purification protects the preparation method of careless biotechnological formulation, it is characterized in that step is by weight:
(1) the high-concentration culturing thing preparation of short lactobacillus: get a ring short lactobacillus CGMCC1.558 lawn from test tube slant, be inoculated in the MRS liquid nutrient medium of 50 ~ 100mL sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 40h; Then accessed 5 ~ 8L in the MRS liquid nutrient medium of sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 35h, made bacterial classification seed liquor for subsequent use; Now viable count>=10 of bacterium liquid
9cFUmL
-1;
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300 ~ 350L, cool for subsequent use after 121 DEG C of sterilizing 15min; By above-mentioned bacterial classification seed liquor access fermentor tank, at 32 ~ 37 DEG C, rotating speed 60 ~ 80rmin
-1condition under cultivate 30 ~ 35h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use, obtain the high-concentration culturing thing of short lactobacillus;
(2) the high-concentration culturing thing preparation of enterococcus faecalis: get a ring enterococcus faecalis CGMCC1.2135 lawn from test tube slant, be inoculated in the MRS liquid nutrient medium of 50 ~ 100mL sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 40h; Then accessed 5 ~ 8L in the MRS liquid nutrient medium of sterilizing, in 32 ~ 37 DEG C of quiescent culture 30 ~ 35h, made bacterial classification seed liquor for subsequent use; Now viable count>=10 of bacterium liquid
9cFUmL
-1;
In 0.5 ton of stainless steel fermentor tank, add the liquid fermentation medium of cumulative volume 300 ~ 350L, cool for subsequent use after 121 DEG C of sterilizing 15min; By above-mentioned bacterial classification seed liquor access fermentor tank, at 32 ~ 37 DEG C, rotating speed 60 ~ 80rmin
-1condition under cultivate 30 ~ 35h, treat bacterial concentration>=10
9cFUmL
-1time the 25L bucket put into through sterilizing for subsequent use, obtain the high-concentration culturing thing of enterococcus faecalis;
(3) preparation of potassium fulvate liquid: get potassium fulvate 25 ~ 40 parts, adds in 65 ~ 80 parts of water, dissolves in 121 DEG C of sterilizing 15min after stirring, for subsequent use after cooling, obtains potassium fulvate liquid;
(4) mix: the high-concentration culturing thing 50 ~ 60 parts getting the obtained short lactobacillus of step (1), the high-concentration culturing thing 35 ~ 45 parts of the enterococcus faecalis that step (2) is worth, the potassium fulvate liquid 5 ~ 10 parts that step (3) is obtained; Above-mentioned mixing of materials is evenly loaded in packing bottle afterwards, obtains product pliotherm period shrimp crab cultivation water purification and protect careless biotechnological formulation.
3. pliotherm period shrimp crab cultivation water purification protects the preparation method of careless biotechnological formulation as claimed in claim 2, it is characterized in that:
Described MRS liquid nutrient medium is, extractum carnis 10gL
-1, peptone 10gL
-1, yeast extract paste 5gL
-1, glucose 5gL
-1, citric acid diamines 2gL
-1, tween 80 1gL
-1, sodium acetate 5gL
-1, dipotassium hydrogen phosphate 2gL
-1, magnesium sulfate 0.2gL
-1, manganous sulfate 0.05gL
-1, calcium carbonate 20gL
-1, adjust ph 6.2 ~ 6.6, cools for subsequent use after 121 DEG C of sterilizing 15min;
Described liquid fermentation medium, its component ratio by mass number meter: 15 parts, molasses, soyflour 5 parts, fresh fish cream 5 parts, yeast extract paste 5 parts, sodium acetate 2 parts, 0.1 part, magnesium sulfate, dipotassium hydrogen phosphate 1.0 parts, 66.9 parts, water, dissolves mixing, regulator solution pH value 6.2 ~ 6.6 under room temperature in fermentor tank, 121 DEG C of sterilizing 15min, for subsequent use after cooling.
4. pliotherm period shrimp crab cultivation water purification protects the preparation method of careless biotechnological formulation as claimed in claim 3, it is characterized in that: sugar content >=55% in described molasses.
5. pliotherm period shrimp crab cultivation water purification protects the preparation method of careless biotechnological formulation as claimed in claim 3, it is characterized in that: the reagent being used for regulating pH is lactic acid and sodium bicarbonate.
6. described in claim 1, pliotherm period shrimp crab cultivation water purification protects the application of careless biotechnological formulation, it is characterized in that: pliotherm period shrimp crab cultivation water purification is protected careless biotechnological formulation and is used in cultivating pool, consumption is 300 ~ 1000g/ mu meter Shui Shen, is used in conjunction 1 ~ 2 time.
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Application publication date: 20151202 |