CN114343087B - Fermented feed for improving prawn culture survival rate and growth speed - Google Patents
Fermented feed for improving prawn culture survival rate and growth speed Download PDFInfo
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
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- A—HUMAN NECESSITIES
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Abstract
The invention relates to a fermented feed for improving the culture survival rate and the growth speed of prawns, which contains a leaven, wherein the leaven comprises the following components: 15-20% of bacillus subtilis, 20-20% of lactobacillus buchneri TZ-LB-01715, 10-15% of enterococcus faecium, 20-25% of saccharomyces cerevisiae, 5-15% of acid protease, 6-12% of xylanase and 5-15% of pectinase; the activity of each probiotic and enzyme preparation was: bacillus subtilis 2X 1010CFU/g, Lactobacillus buchneri TZ-LB-0172X 1010CFU/g, enterococcus faecium 1X 1010CFU/g, Saccharomyces cerevisiae 2X 109CFU/g, acid protease 5X 104U/g, xylanase 1X 104U/g, pectinase 3X 103U/g. The leaven has obvious bacteriostasis effect on vibrio parahaemolyticus, can promote the growth of prawns and obviously reduce the early death of young prawns.
Description
Technical Field
The invention relates to the technical field of feeds, in particular to a fermented feed for improving the culture survival rate and the growth speed of prawns.
Technical Field
With the large-scale and intensive development of aquaculture in China, the environment of aquaculture water is seriously polluted, the disease incidence rate of aquaculture is continuously improved, and the product quality of aquaculture is also continuously reduced. Although the addition of antibiotics to feed has been prohibited, the long-term use of antibiotic feed has previously affected the development of resistance of aquatic animals to pathogenic bacteria and harmful microorganisms. The problem of how to solve the disease of aquatic animals is a major problem facing the current aquaculture industry in case antibiotics cannot be used.
The penaeus vannamei boone is an absolute dominant variety in aquaculture in China. The penaeus vannamei boone is originally produced in the coastal water area of the south pacific ocean and is tropical economic shrimps, and through continuous exploration, research and test of researchers, the penaeus vannamei boone seedlings can be cultured in a large amount in fresh water after being gradually desalted. However, due to the abuse of antibiotics and the pollution of water quality in recent years, the penaeus vannamei boone has a plurality of diseases in the culture process, and huge economic losses are brought to farmers. It has been shown that common diseases include: white spot syndrome, prawn red body disease, muscle necrosis (white turbidity) disease, red leg disease, yellow gill disease, gill rot disease, etc. It is worth noting that the most common outbreak of the prawn early death syndrome is related to vibrio parahaemolyticus, and has the characteristics of wide disease scope, fast disease speed and unobvious disease season; the early stage of symptoms is red leg and tail, reddish liver and pancreas, broken beard of red hair, and reddish intestine and stomach, and the later stage is marked by atrophy of jejunum, empty stomach, liver and pancreas, reddish body, and immobility.
The Vibrio parahaemolyticus is also called halophilic bacteria, and the growth conditions are suitably 37 ℃ and pH 7.4-8. The vibrio parahaemolyticus has wide distribution, mainly lives in seawater, fishes, shrimps and shell crustaceans, usually causes gastrointestinal infection, and can also cause parenteral infection. In recent years, the pathogenic bacteria of the early death syndrome of penaeus vannamei boone are mainly vibrio parahaemolyticus. In the culture of the prawns, the body color of the sick prawns is whitish, turbid and reddish, most of the liver and pancreas are swollen, the body is soft, the color is pale white or faint yellow, and the liver and pancreas of part of the sick prawns are obviously atrophied. Usually, the disease condition develops rapidly with slow flowing of pond water or lying on the slope of the pond, losing appetite and jejunum and stomach, the mortality and the pond discharging rate are extremely high, and the shortest time is only 2 to 3 days from a small amount of disease to the pond discharging.
The pathogenic mechanism of the vibrio parahaemolyticus is that the vibrio parahaemolyticus generates hemolytic toxin to cause hemolysis of various cells of the prawn; vibrio parahaemolyticus has enterotoxin effect, which causes prawn digestive tract tissue injury, ileum erosion, gastric mucositis, even liver, pancreas and digestive dysfunction; the vibrio parahaemolyticus generates urease, so that intestinal juice of the prawns is accumulated to generate intestinal dropsy, and the phenomenon of white feces can occur in severe cases; the extracellular product of vibrio parahaemolyticus can obviously reduce the immunity of prawns and has slow growth speed, so that various other diseases are easily caused.
The biological fermentation feed is a feed capable of replacing feed added with antibiotics. The feed is added with viable bacteria preparation, plant extract, organic acid, antibacterial peptide, etc., and the health of the product is far higher than that of the feed added with antibiotic. The microorganisms in the feed have a metabolic effect, can generate small molecular substances such as organic acid, soluble polysaccharide and the like by degrading polysaccharide, protein, fat and the like, can promote absorption, contains rich nutrient substances, has good palatability, and has a promoting effect on improving the problem of water pollution of aquaculture. However, no biological fermentation feed for preventing and treating the vibrio parahaemolyticus of the penaeus vannamei exists in the current market.
Disclosure of Invention
In order to solve the problems, the invention provides a fermented feed for improving the culture survival rate and the growth speed of prawns.
The improvement provided by the inventionThe shrimp culture survival rate and growth speed fermented feed contains a leaven, and the leaven comprises the following components: 15-20% of bacillus subtilis, 20-20% of lactobacillus buchneri TZ-LB-01715, 10-15% of enterococcus faecium, 20-25% of saccharomyces cerevisiae, 5-15% of acid protease, 6-12% of xylanase and 5-15% of pectinase; wherein the activities of the probiotics and the enzyme preparation are respectively as follows: bacillus subtilis 2X 1010CFU/g, Lactobacillus buchneri TZ-LB-0172X 1010CFU/g, enterococcus faecium 1X 1010CFU/g, Saccharomyces cerevisiae 2X 109CFU/g, acid protease 5X 104U/g, xylanase 1X 104U/g, pectinase 3X 103 U/g。
Wherein the Lactobacillus buchneri TZ-LB-017 is classified and named as Lactobacillus buchneriLactobacillus buchneriThe culture medium is preserved in China general microbiological culture Collection center (CGMCC), and the preservation addresses are as follows: the preservation time is 2021 year, 3 month and 23 day, and the preservation number is CGMCC No. 22054.
Further, the preparation method of the fermented feed comprises the following steps: 0.05-0.2Kg of leaven, 50-200Kg of basic raw materials, 5-50Kg of brown sugar and 10-50Kg of water, evenly stirring after mixing, and fermenting for three days in a closed manner at normal temperature.
Further, the base stock comprises: 10-20% of rapeseed cake, 45-55% of soybean meal, 16-26% of fish meal, 5-20% of bran and 5-20% of corn protein powder.
Preferably, the base stock comprises: 15% of rapeseed cake, 50% of soybean meal, 20% of fish meal, 8% of bran and 7% of corn protein powder.
The invention has the following beneficial effects:
1. the lactobacillus buchneri TZ-LB-017 screened from pig manure of a farm by the inventor is adopted in the starter, and compared with the conventional lactobacillus buchneri or the commercially available lactobacillus buchneri, the lactobacillus buchneri can generate phenyllactic acid with higher concentration in the fermentation process, and has remarkable advantages in the aspects of acid production, harmful bacteria inhibition and synergistic effect with other probiotics. The invention discovers that the leavening agent obtained by compounding lactobacillus buchneri TZ-LB-017 with other bacteria and enzymes has obvious bacteriostatic effect on vibrio parahaemolyticus: the plate dibbling method of example 1 proves that the leavening agent of the present invention has bacteriostatic effect on vibrio parahaemolyticus compared with the common leavening agent.
2. The fermented feed prepared by the leavening agent has obvious improvement effect on the survival rate, the growth speed, the feeding liveness and the bait efficiency of the penaeus vannamei boone, which shows that the fermented feed prepared by the leavening agent can obviously improve the species and the number of dominant flora fixedly planted in the intestinal tract of the penaeus vannamei boone, improve the immunity of the penaeus vannamei boone, obviously improve the disease resistance of vibrio parahaemolyticus, obviously reduce the early death condition of the penaeus vannamei boone, improve the health state of the penaeus vannamei boone, and improve the economic benefit of the penaeus vannamei boone cultivation without antibiotics in the feed in production.
Drawings
FIG. 1 is a table-type dibbling method of example 1, which verifies that the leavening agent of the present invention has bacteriostatic effect on Vibrio parahaemolyticus;
FIG. 2 is a comparison of 45 days of Penaeus vannamei Boone in the large herd breeding trial of example 3.
Detailed Description
The present invention is further illustrated by the following examples.
Example 1: bacteriostatic test of leavening agent
1.1 preparation of culture Medium and leaven
Liquid culture medium: to 100 ml of filtered seawater were added 0.5g of peptone (Typtone), 0.1 g of Yeast Extract (Yeast Extract), and 0.001 g of iron phosphate tetrahydrate (Fe PO)4•4H2O), adjusting the pH value to 7.6-7.8, sterilizing at high temperature and high pressure (121 ℃, 20 min) by a high-pressure steam sterilizer, and storing in a chromatography cabinet at 4 ℃ for later use.
Solid medium: to 100 ml of filtered seawater were added 0.5g of peptone (Typtone), 0.1 g of Yeast Extract (Yeast Extract), 0.001 g of iron phosphate tetrahydrate (Fe PO)4•4H2O), 2g Agar (Agar Powder), sterilized by high-pressure steam sterilizer at high temperature and high pressure (121 deg.C, 20 min), and inverted in a sterile ultra-clean bench when the temperature of the culture medium is reduced to about 50 deg.C, each plateAbout 15 ml, after air drying, it is stored in a chromatography cabinet at 4 ℃ for later use.
A leavening agent: the components of the leavening agent comprise: 15-20% of bacillus subtilis, 20-20% of lactobacillus buchneri TZ-LB-01715, 10-15% of enterococcus faecium, 20-25% of saccharomyces cerevisiae, 5-15% of acid protease, 6-12% of xylanase and 5-15% of pectinase; wherein the activities of the probiotics and the enzyme preparation are respectively as follows: bacillus subtilis 2X 1010CFU/g, Lactobacillus buchneri TZ-LB-0172X 1010CFU/g, enterococcus faecium 1X 1010CFU/g, Saccharomyces cerevisiae 2X 109CFU/g, acid protease 5X 104U/g, xylanase 1X 104U/g, pectinase 3X 103 U/g。
Wherein the Lactobacillus buchneri TZ-LB-017 is classified and named as Lactobacillus buchneriLactobacillus buchneriThe culture medium is preserved in China general microbiological culture Collection center (CGMCC), and the preservation address is as follows: the preservation time is 2021 year, 3 month and 23 day, and the preservation number is CGMCC No. 22054.
1.2 test methods
Conventional leavening agent: 10% of bacillus subtilis, 9% of lactobacillus plantarum, 30% of saccharomyces cerevisiae, 10% of enterococcus faecalis, 30% of cellulase and 11% of xylanase.
2g of the leaven and the conventional leaven are respectively taken, added with a small amount of filtered seawater for activation, and then respectively transferred into conical flasks filled with 200 ml of liquid culture medium, and cultured in a constant temperature incubator at 28 ℃ for 12 hours for later use.
A strain of vibrio parahaemolyticus III-1 preserved in a laboratory is taken as an indicator bacterium, and the activated vibrio parahaemolyticus is transferred into an erlenmeyer flask filled with 100 ml of liquid culture medium for culture overnight.
100 mu l of the III-1 bacterial liquid is taken and coated on a solid culture medium, then 4 aseptic filter paper sheets with the r = 6 mm are clamped by aseptic tweezers and placed on a flat plate coated with the indicator bacteria at equal distance, 8 mu l of leaven is respectively dropped on two filter paper sheets (P1/P2), E1 and E2 are conventional leaven, and the flat plate is taken after 24 h to observe whether bacteriostatic rings appear around the filter paper sheets.
1.3 test results
As shown in figure 1, the experimental result of the plate dibbling method shows that compared with the common leaven, the leaven of the invention has antagonistic effect on vibrio parahaemolyticus.
Example 2 laboratory culture test of prawn
Preparing a fermented feed: 0.1Kg of leaven, 150Kg of basic raw materials, 25Kg of brown sugar and 25Kg of water, evenly stirred and fermented in a closed way for three days at normal temperature. The base material comprises: 15% of rapeseed cake, 50% of soybean meal, 20% of fish meal, 8% of bran and 7% of corn protein powder.
Randomly distributing 840 tails of penaeus vannamei boone larvae into 6 water bodies of 0.4 m3Of the indoor round cement pit, at the end of each pit 140. The breeding is temporarily carried out for 15 days before formal group feeding is started, and the pathogens possibly carried by the breeding are sampled and detected during the temporary breeding. The test is provided with a blank control group and a test group, and each group is provided with 3 parallels. The test prawn fry is fed for 3 times (8 o ' clock, 2 o ' clock in afternoon and 8 o ' clock in evening) every day, and the feeding amount is 3% -5% of the weight of the prawn fry (the feeding amount increases along with the culture time). Wherein the blank control group is fed with common feed; the test group is fed with fermented feed in the first 30 days and common feed in the last 30 days in a period of 60 days. The temperature of the water body is 24-27 ℃, the salinity of the water body is 15 (+ -1), the pH value is 8.1 (+ -0.1), the air is continuously filled, the ingestion and survival conditions of the young prawns are checked every day, and the dead prawns are taken out in time.
Measuring the initial weight and the weight of each group of prawn seedlings at the end of the test, and calculating the specific growth rate of each group of prawn seedlings; calculating the survival rate of the prawns at the end of the test;
survival (%) = survival mantissa/total mantissa × 100%;
the results show that: the survival rate of each group of prawns in 60 days of culture is only (25.5 +/-1.8)%, while the survival rate of prawns fed with the fermented feed reaches (79.3 +/-7.6)%, and the difference is very obvious (P)<0.01)。
Example 3 aquatic Mass Breeding test
Fermented feed was prepared as in example 2.
The test is carried out in Suzhou city of Jiangsu province, the test pond is 4 prawn greenhouse culture ponds, the pond area is 5 mu, and the bottom materials and the hydrological conditions of the ponds are completely the same. No. 1 and No. 3 ponds are used as test groups, and biological fermentation feed is continuously fed with the bait (the feeding proportion of the fermentation feed to the common feed is shown in a table 2); the No.2 and No. 4 ponds are used as control groups and are fed with common feed in the whole process. In the whole test process, the conventional culture operation of 4 shrimp ponds is the same, the average temperature in the greenhouse is 23.8 ℃ in the test period, and the test period is 60 days. The test is carried out by selecting shrimp fries with the same specification, size and health condition.
Measurement indexes are as follows: the growth condition of the shrimp larvae: every 15 days in the test, 50 shrimps were randomly taken from each shrimp pond, observed for the body color, intestinal tract and liver and pancreas shape and color, and measured for body length and weight and recorded. At the end of the test, the harvested test shrimps were measured for body length, body weight and recorded.
Feeding time: 1 material platform is placed respectively at diagonal position in every pond, and the fodder volume is placed according to 1% of the total amount of feeding when having a meal in the material platform, observes every day and records the time that the fodder eats futilely in the material platform.
Bait efficiency: the bait feeding amount is recorded every day, and the total feeding amount and the shrimp receiving amount are calculated at the end of the test.
The results show that: the test group shrimp larvae are transparent in body color, the number of black spots on the head, chest and nails is small, the intestinal tract is full, the liver and pancreas outlines are clear, the color is reddish brown, and the length of excrement is normal and is not easy to break. The contrast group shrimps are darker, part of the shrimps are reddish, more black spots are formed on the shells, stress reaction is easy to generate and twitch, the intestinal tract is thinner, the liver and pancreas are clear in outline, the volume is larger, the color is reddish, and the length of the prawn excrement is short and the color is dark.
3.1 growth rate: as can be seen from Table 3 and FIG. 2, the growth rate of the prawns was significantly higher in the test group fed with the fermented feed than in the control group.
3.2 bait efficiency and survival rate
The feeding time of the test group prawns is found to be shortened by counting the feeding time of the prawns every day, which indicates that the prawns have active feeding behaviors, and simultaneously proves that the feeding attraction effect of the fermented feed is obvious. After 60 days of culture, 3401Kg of prawns are harvested in two ponds (No. 1 and No. 3 ponds) of a test group, and the bait efficiency reaches 1.14; and 2773 Kg of prawns are harvested from two ponds (No. 2 and No. 4) of the control group together, and the bait efficiency is 1.03. The survival rate of the shrimps of each test group is calculated according to the yield of the shrimps and the average weight of the monomers, which shows that the survival rate of the fermented feed can be improved by 3.1 percent, and the bait efficiency is improved by 0.11.
The survival rate of the shrimp seedlings in the test group fed with the biological fermented feed is 67.1 percent, and the economic benefit is 6134 yuan/mu (see table 4), while the survival rate of the shrimp seedlings in the control group is 64.0 percent, and the economic benefit is 5351 yuan/mu.
The tests prove that when the feed fermented by the starter is fed in the breeding process of the penaeus vannamei boone, the types and the number of dominant flora fixedly planted in intestinal tracts of the penaeus vannamei boone can be obviously improved, the immunity of the penaeus vannamei boone is improved, the disease resistance to vibrio parahaemolyticus is obviously improved, the early death condition of the young penaeus vannamei boone is obviously reduced, the health state of the penaeus vannamei boone is improved, the body color of the young penaeus vannamei boone in a test group is transparent, the ingestion time is shortened, the growth of the young penaeus vannamei boone can be obviously promoted, the survival rate of the young penaeus vannamei boone and the bait efficiency are improved, and the economic benefit of farmers is obviously improved.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (4)
1. A fermented feed for improving the culture survival rate and the growth speed of prawns, which contains a leaven, is characterized in that the leaven comprises the following components: 15-20% of bacillus subtilis, 20-20% of lactobacillus buchneri TZ-LB-01715, 10-15% of enterococcus faecium, 20-25% of saccharomyces cerevisiae, 5-15% of acid protease, 6-12% of xylanase and 5-15% of pectinase; wherein the activities of the probiotics and the enzyme preparation are respectively as follows: bacillus subtilis 2X 1010CFU/g, Lactobacillus buchneri TZ-LB-0172X 1010CFU/g, enterococcus faecium 1X 1010CFU/g, Saccharomyces cerevisiae 2X 109CFU/g, acid protease 5X 104U/g, xylanase 1X 104U/g, pectinase 3X 103 U/g;
Wherein the Lactobacillus buchneri: (A)Lactobacillus buchneri) TZ-LB-017 and is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation time is 2021, 3 and 23 days, and the preservation number is CGMCC No. 22054.
2. The fermented feed for improving the cultivation survival rate and the growth speed of prawns according to claim 1, wherein the preparation method of the fermented feed comprises the following steps: 0.05-0.2Kg of leaven, 50-200Kg of basic raw materials, 5-50Kg of brown sugar and 10-50Kg of water, evenly stirring after mixing, and fermenting for three days in a closed manner at normal temperature.
3. The fermented feed for improving the cultivation survival rate and the growth speed of prawns according to claim 2, wherein the basic raw materials comprise: 10-20% of rapeseed cake, 45-55% of soybean meal, 16-26% of fish meal, 5-20% of bran and 5-20% of corn protein powder.
4. The fermented feed for improving the survival rate and the growth speed of the prawn culture according to claim 3, wherein the basic raw materials comprise: 15% of rapeseed cake, 50% of soybean meal, 20% of fish meal, 8% of bran and 7% of corn protein powder.
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