CN111763644B - Composite microbial agent for preventing and treating liver and gall syndrome of fish as well as preparation method and application thereof - Google Patents

Composite microbial agent for preventing and treating liver and gall syndrome of fish as well as preparation method and application thereof Download PDF

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CN111763644B
CN111763644B CN202010723522.XA CN202010723522A CN111763644B CN 111763644 B CN111763644 B CN 111763644B CN 202010723522 A CN202010723522 A CN 202010723522A CN 111763644 B CN111763644 B CN 111763644B
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bacillus amyloliquefaciens
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刘刚
顾欣燕
刘金龙
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Tianjin Development Zone Kunhe Biotechnology Co ltd
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Abstract

The invention relates to a compound bacterial agent for preventing and treating fish liver and gall syndrome, and a preparation method and application thereof, wherein the compound bacterial agent is composed of bacillus coagulans and bacillus amyloliquefaciens in scientific proportion, and the bacillus coagulans and the bacillus amyloliquefaciens cooperate to have obvious treatment effect on liver and gall syndrome; the composite microbial inoculum has the characteristics of no phytotoxicity, no residue, growth promotion and small dosage, and provides possibility for diversification of medication schemes and protection of ecological environment.

Description

Composite microbial agent for preventing and treating liver and gall syndrome of fish as well as preparation method and application thereof
Technical Field
The invention belongs to the field of microbial inoculant products, and particularly relates to a compound inoculant for preventing and treating fish liver and gall syndrome, and a preparation method and application thereof.
Background
Liver and gall syndrome is one of the most frequent fish diseases in the occurrence of fish diseases in recent years, the epidemic season mainly starts from 6 months and the occurrence of 10 months, the disease incidence rate of fries and fingerlings is high in all places throughout the country, and the harmful objects are carp, crucian carp, grass carp, channel catfish, snakehead, rainbow trout, split-belly fish, megalobrama amblycephala, black carp and tilapia, and the disease is also common in soft-shelled turtles. As the common misdiagnosis is that other fish diseases are treated, the medicine is not symptomatic, and the death rate can reach 50 to 60 percent, even 60 to 90 percent.
Hepatobiliary syndrome is a typical symptom of hepatobiliary enlargement and discoloration. In the early stage of disease of the diseased fish, the liver is slightly enlarged, the liver is slightly anemic, and the color is slightly pale; the gallbladder is darker and slightly greenish. Along with the development of the disease, the liver is obviously enlarged, can be more than 1 time larger than that of the normal condition, the liver is gradually yellow and whitened, or is in plaque-like yellow-red-white phase to form obvious liver-pattern symptoms, some liver is changed into 'green liver' partially or mostly, some liver is slightly touched and fragile, the gall bladder is obviously enlarged by 1-2 times (sometimes causes bile overflow or gall bladder rupture), the gall color is changed into dark green or dark green, or is changed into yellow and white until colorless, the gall of a heavy patient is engorged with redness, and the gall is also changed into red. Sometimes, the spleen and kidney of the diseased fish are also obviously enlarged, and the viscera system is enlarged at the same time. As the main viscera are severely diseased and damaged, the disease resistance of the organism is reduced, and the invasion of other germs is allowed, the serious patients often have bleeding, gill rot, enteritis, tail rot and other symptoms. The most typical characteristic of this disease is hepatobiliary enlargement and discoloration.
Liver and gall are organs for decomposing toxins, all toxic substances in water and various toxins (mycotoxins and the like) in feed are detoxified through fish livers, and all medicines almost increase the burden of livers.
From the current research results, we find that the causes of hepatobiliary syndrome are mainly three types:
1. is the deterioration of the culture density and the water environment. When the ammonia nitrogen concentration in the water body is too high, the metabolic products of ammonia in the fish body are difficult to normally discharge and accumulate in the blood, and the metabolic imbalance of the fish is caused to cause liver and gall diseases.
2. Is a medicament for disorder use. For example, furazolidone, chloramphenicol, sulfonamides and the like are added into the feed at low dosage for a long time, or the drugs with large side effects and high residues such as trichlorfon, copper sulfate and the like are used, so that liver injury is caused.
3. Is the spoilage of the feed, unbalance of nutrient components and deficiency of toxic substances and vitamins contained in the feed. Such as too high protein content, too high carbohydrate content, or long-term use of animal fats and highly saturated fatty acids, etc., results in too high feed energy-to-protein ratios. The high protein feed is easy to induce liver fat accumulation, destroy liver function and interfere normal physiological and biochemical metabolism of fish. The excessive carbohydrate content can cause disorder of sugar metabolism of fish, cause accumulation of visceral fat and prevent normal functions, the main lesion part is liver, a great amount of liver sugar is accumulated and fat infiltrates, hepatomegaly, light color and luster are caused, the appearance is matt, and serious fatty liver can also cause liver lesion to lose normal functions of liver, so that fish diseases which are mainly characterized by liver and gall diseases, namely, liver and gall syndrome of fish, appear.
Oxidation, rancidity, mildew and deterioration of fish feed. Fat is a substance which is easy to oxidize, and aldehyde, ketone and acid generated by fat oxidation are toxic to fish and directly cause damage to liver.
Choline, VE, biotin, inositol, VB and the like are all involved in fat metabolism in fish bodies, and the lack of the vitamins can cause fat metabolism disorder in fish bodies, so that fat is accumulated in liver and liver diseases are induced.
The gossypol in the cotton dregs, the glucosinolate in the vegetable dregs, the nitrite in the inferior fish meal and other toxic and harmful substances can cause liver and gall diseases of fish.
The current methods for treating hepatobiliary syndrome include the following methods:
1. adding bile acid
Exogenous bile acid can directly enter liver and gall to improve the utilization of grease and fat-soluble vitamins by aquatic animals, but can also cause the function degradation of the secretion bile of liver and pancreas, and is more seriously dependent on exogenous bile acid.
2. Scientific feeding and reduction of cultivation density
The scientific feeding contains the feeding stuff which does not deteriorate (without various toxins), and the protein content and the carbohydrate nutrition in the feeding stuff just meet the growth requirement of fish, and various vitamins have no more elements and no less elements. Scientific feeding also comprises moderate feeding quantity, and liver burden of fishes cannot be caused by excessive feeding. The scientific feeding and the reduction of the cultivation density have the consequences of reducing the investment of feed and the cultivation quantity in unit area, and the two modes reduce the production efficiency. Is unfavorable for the development of intensive cultivation.
3. Proper administration of drugs
Proper administration is to reduce unnecessary antibiotics and to avoid prohibiting the type of drug used in aquaculture. However, the use of antibiotics is reduced, and the disease loss is increased.
4. Supplementing vitamins and microelements
The starting point of supplementing vitamins and microelements is good, but as a common farmer and a small-sized feed factory, the method can hardly use N multiple vitamins and microelements accurately for each pond and multiple fish in mixed culture. Once the grasping is inaccurate, the nest falls into excess and deficiency.
Through continuous researches of technicians of the company, no report of preventing and treating hepatobiliary syndrome by using bacillus coagulans and bacillus amyloliquefaciens is available.
Disclosure of Invention
The invention aims to provide a compound microbial inoculum for preventing and treating fish liver and gall syndrome, and a preparation method and application thereof. The composite microbial inoculum is composed of bacillus coagulans and bacillus amyloliquefaciens in scientific proportion, and the bacillus coagulans and the bacillus amyloliquefaciens have a synergistic effect and obvious treatment effect on hepatobiliary syndrome; the composite microbial inoculum has the characteristics of no phytotoxicity, no residue, growth promotion and small dosage, and provides possibility for diversification of medication schemes and protection of ecological environment.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a compound microbial agent for preventing and treating fish liver and gall syndrome is characterized in that: including bacillus coagulans and bacillus amyloliquefaciens;
the bacillus coagulans (Bacillus coagulans) MES847, date of deposit: 8.30 days of 2018, the name of the preservation unit and short: china general microbiological culture collection center (CGMCC), number of collection: 16358; bacillus subtilis (Bacillus coagulans) MES847 is from the same sources as in patent application Ser. No.: a method for ecologically culturing Litopenaeus vannamei by replacing partial feed with live baits.
The bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MES 812 is preserved in China general microbiological culture collection center (China general microbiological culture Collection center) with a preservation address of Hospital No. 3 of North Chen West Lu 1 of the Korean area of Beijing, and a preservation date of 2017, 8, 10 days and a preservation number of CGMCC No.14515; the source of the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MES 812 is the same as the patent application number: CN201710947311.2 is a method for preventing and controlling dead tree of medlar root rot.
Effective activity of bacillus amyloliquefaciens in composite microbial inoculumThe number of bacteria is not less than 1.0X10 10 The effective viable count of bacillus coagulans per gram is not less than 5.0X10 8 Each/g.
Preferably, after the bacillus coagulans and the bacillus amyloliquefaciens are respectively cultured, bacterial powder is prepared, and then the bacterial powder and the bacterial powder are mixed according to a proportion to prepare the composite bacterial agent.
Preferably, the mixed ratio of the bacillus coagulans bacterial powder and the bacillus amyloliquefaciens bacterial powder in the composite bacterial agent is 1:1 by weight.
The invention also provides a preparation method of the compound microbial inoculum for preventing and treating fish hepatobiliary syndrome, which comprises the following steps: the bacillus coagulans and the bacillus amyloliquefaciens are subjected to shake flask culture, primary seed culture, secondary seed culture, fermentation, concentration and spray drying respectively to prepare bacterial powder, and then are mixed according to a proportion to prepare the composite bacterial agent.
Preferably, the culture method of the bacillus coagulans (Bacillus coagulans) MES847 is the same as that of patent CN201911419440.X, namely a method for ecologically culturing Litopenaeus vannamei by replacing part of feed with live baits;
the culture method of the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MES 812 is the same as that of patent application number: CN201710947311.2 is a method for preventing and controlling dead tree of medlar root rot.
The invention also provides application of the composite microbial inoculum for preventing and treating the liver and gall syndrome of fish, wherein the composite microbial inoculum is used for preventing the liver and gall syndrome of fish: the compound microbial inoculum is used in the fish feed according to the mass ratio of 1000:2, and is continuously added in the middle and later periods of cultivation to prevent liver and gall syndrome of fish.
Preferably, the compound bacterial agent is used for treating liver and gall syndrome of fish: the composite microbial inoculum is used in fish feed, the mass ratio of the fish feed to the composite microbial inoculum is 1000:4, and the fish feed is continuously added for 10-15 days to treat liver and gall syndrome of fish.
The principle of the invention:
the bacillus coagulans MES847 has a relatively wide antibacterial spectrum and has the effects of improving the cellular immunity and humoral immunity level of organisms; can stimulate the development of animal immune organs and improve the disease resistance of animals; active substances such as protease, lipase, amylase and xylanase can be produced, and digestion and absorption of feed nutrient substances (especially redundant fat) are promoted; vitamins, short chain fatty acids, amino acids, growth promoting factors and the like which are produced can provide nutrition for animal organisms, promote peristalsis of small intestines, improve intestinal digestion function and provide favorable conditions for growth and development of animals; the organic acid produced may act as part of the bile acid.
Active substances such as polymyxin, nystatin, gramicidin and the like generated in the growth process of bacillus amyloliquefaciens MES 812 have obvious inhibition effect on pathogenic bacteria or endogenous infected conditional pathogenic bacteria, and the morbidity is reduced; stimulating the growth and development of animal immune organs, activating T, B lymphocytes, improving the level of immunoglobulin and antibody, enhancing cellular immunity and humoral immunity functions, improving group immunity and reducing medication; bacillus amyloliquefaciens thallus synthesizes alpha-amylase, proteinase, lipase, cellulase and other enzymes and acts together with animal digestive enzymes in digestive tract; can synthesize various B vitamins such as vitamin B1, vitamin B2, vitamin B6, nicotinic acid and the like, and improve the activity of interferon and macrophage in animals; bacillus amyloliquefaciens is facultative anaerobe, rapidly consumes free oxygen in intestinal tracts, causes intestinal hypoxia, promotes the growth of beneficial anaerobes, and indirectly inhibits the growth of other pathogenic bacteria.
The two bacterial powders have different functions for the etiology of different hepatobiliary syndromes, and solve hepatobiliary syndromes caused by almost all reasons.
Advantageous effects
The composite microbial inoculum has the characteristics of no phytotoxicity, no residue, growth promotion and small dosage, and provides possibility for diversification of medication schemes and protection of ecological environment.
Compared with the method for adding cholic acid juice, the organic acid generated by bacillus coagulans in the compound bacterial agent can be used as endogenous organic acid, and the bacterial strain has self-regulation, and when the acid yield reaches a certain level, the reproduction is stopped, and the organic acid is not generated any more, so that the recovery of bile acid secretion of fish livers and gall is facilitated.
Compared with a scientific feeding method, the composite microbial inoculum is added to perfectly solve the relations of feeding quantity, slow growth, culture density and fish yield. The bacillus amyloliquefaciens thalli in the composite microbial inoculum synthesizes alpha-amylase, protease, lipase, cellulase and other enzymes, and plays roles together with digestive enzymes in animal bodies in the digestive tract; promote the absorption of redundant nutrition and reduce the detoxification pressure of the liver.
Compared with the correct medication, the active substances such as polymyxin, nystatin, gramicidin and the like generated in the growth process of the bacillus amyloliquefaciens in the compound bacterial agent are added, and the active substances have obvious inhibition effect on pathogenic bacteria or endogenous infection conditional pathogenic bacteria, reduce morbidity and reduce detoxification pressure of livers when the compound bacterial agent is used for medication.
Compared with the method of supplementing vitamins and microelements, the bacillus amyloliquefaciens added into the compound microbial inoculum can synthesize various B vitamins such as vitamin B1, B2, B6, nicotinic acid and the like, and improve the activity of interferon and macrophages in animals. And the added vitamins and the living bacteria are different, because the added vitamins can be lost in the intestinal tract, and the living bacteria can be produced more efficiently (without loss) and more permanently.
Detailed Description
The invention is further described below in conjunction with the detailed description.
The culture method of bacillus coagulans (Bacillus coagulans) MES847 is the same as that of patent CN201911419440.X, which is a method for ecologically culturing Litopenaeus vannamei by replacing partial feed with live baits;
the culture method of the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MES 812 is the same as that of patent application number: CN201710947311.2 is a method for preventing and controlling dead tree of medlar root rot.
Example 1
A preparation method of a compound microbial inoculum for preventing and treating fish liver and gall syndrome comprises the following steps: bacillus coagulans MES847 and Bacillus amyloliquefaciens MES 812 are subjected to shake flask culture, primary seed culture, secondary seed culture and fermentation respectivelyConcentrating, spray drying to obtain bacterial powder, and mixing proportionally to obtain the composite bacterial agent; the mixed proportion of the bacillus coagulans bacterial powder and the bacillus amyloliquefaciens bacterial powder in the composite bacterial agent is 1:1; the effective viable count of Bacillus amyloliquefaciens in the composite microbial inoculum is not less than 1.0X10 10 The effective viable count of bacillus coagulans per gram is not less than 5.0X10 8 Each/g.
Example 2
The compound bacterial agent for preventing and treating the liver and gall syndrome of the fish prepared in the embodiment 1 of the invention is used for treating the liver and gall syndrome of the fish,
test of therapeutic effect for preventing and treating liver and gall syndrome of fish
(1) Test time and place:
the test is carried out in 2 open-air ponds in Wang Cun fishing ground of temple of Tianjin city at 7 months of 2019, the areas of the 2 ponds are respectively 30 mu (1#) and 50 mu (2#), and the average water depth of the ponds is 1.8 m during the test.
(2) Test pond conditions:
the 2 ponds are mixed fish ponds mainly comprising carps, and the seedling placing, feeding and water feeding conditions of the two ponds are basically consistent. Before the start of the experiment, slow feeding occurred in all 2 ponds, and some of the fish in pond # 2 died. The main cultured fish (carp) is anatomic and inspected to find out the hepatobiliary enlargement, the color change of the gall bladder is darker and the color is slightly green. Both ponds have bacterial gill rot and slight enteritis symptoms. Some fish in pond # 2 appear liver color gradually turning yellow and white.
(3) The test method comprises the following steps:
no.1 is a control pond using commercially available florfenicol powder (for aquatics) and povidone-iodine solution (for aquatics). The 2# pond is a test pond, and the hepatobiliary syndrome is more serious than the 1# pond.
The 7 month 28 day trial began with 400g of the complex inoculant per 100kg of feed in the 2# pond feed, i.e., 4kg of the complex inoculant per ton of compound feed. And the addition was continued for up to 8 months 12 days. And the day was splattered outside the pond # 2: povidone-iodine solution (for aquatic products).
(4) Test results:
the 2# experimental pond gradually tended to be normal in fish school feeding on day 6 (day 8, 3). Dissections showed that liver and gall color gradually changed to normal, and dissections showed that liver and gall size gradually changed to normal by 8 months and 12 days.
The fish in the # 1 control pond on day 6 instead died and on day 7 (day 4 of 8 months) were treated according to the experimental (# 2) pond method. After 5 days (8 months and 9 days), the ingestion of the fish in the pond is also recovered to be normal. And the liver and gall size is gradually changed into normal after 3 days (8 months and 12 days) of dissection.
(5) Conclusion of the test:
therefore, the microecological composite microbial agent obtained by the invention has obvious treatment effect on hepatobiliary syndrome.
Example 3
The prevention experiment effect of different bacterial agents on the liver and gall syndrome of fish:
(1) Test time and place:
the method is carried out in 3 open-air ponds of a new village fishing ground built in Tianjin city Wang Wenzhuang, wherein the areas of the 2 ponds are respectively 50 mu (1#), 30 mu (2#), and 40 mu (3#), and the average water depth is 1.8 m during the test.
(2) Test pond conditions:
of the 3 ponds, pond 1 is a carp-based mixed culture pond, and two ponds 2 and 3 are grass carp-based mixed culture ponds. The seedling placing, feed feeding, and water feeding conditions of the 3 ponds are basically consistent. Symptoms of hepatobiliary syndrome appear in all 3 ponds before the start of the test.
(3) Test protocol:
the pond 1 uses a single bacillus amyloliquefaciens bacterial agent, the pond 2 uses a single bacillus coagulans bacterial agent, and the pond 3 uses a composite bacterial agent to treat hepatobiliary syndrome. And comparing the recovery conditions of the hepatobiliary syndromes of the ponds to obtain a conclusion.
(4) The test process comprises the following steps:
the test starts at 15 days of 2019, 6 months, 400g of the composite microbial agent is used in 3# pond feed per 100kg of feed, namely 4kg of the composite microbial agent is used per ton of compound feed; 400g of bacillus coagulans microbial inoculum is used in the No. 2 pond feed per 100kg of feed, namely 4kg of the microbial inoculum is used per ton of compound feed; in the pond feed No.1, 400g of bacillus amyloliquefaciens microbial inoculum is used per 100kg of feed, namely 4kg of microbial inoculum is used per ton of compound feed. All three ponds were added and continued for up to 6 months and 30 days.
The 3# experimental pond gradually tended to be normal in fish school feeding on day 6 (21 days of 6 months). Dissections found that the color of the liver and pancreas gradually turned to normal, and dissections found that the size of the liver and pancreas gradually turned to normal by 6 months and 30 days.
Fish feeding gradually became normal on day 8 (day 23 of 6 months) of the trial pond # 1. Dissections found that the color and size of the liver pancreas gradually changed to normal by 7 months and 5 days.
Fish feeding gradually became normal on day 8 (day 23 of 6 months) of the 2# trial pond. Dissections found that the color and size of the liver pancreas gradually changed to normal by 7 months and 5 days.
(5) Test results:
the pond # 3 heals at the fastest rate with minimal loss. And the healing speed of the pond 1 and the pond 2 is obviously slower than that of the pond 3, and the ingestion time is influenced to be longer.
Therefore, the bacillus amyloliquefaciens and the bacillus coagulans have a synergistic effect, and the combined use has a better effect of preventing and treating hepatobiliary syndrome.
The foregoing is merely a preferred embodiment of the invention, and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended by the present invention.

Claims (3)

1. A compound microbial agent for preventing and treating fish liver and gall syndrome is characterized in that: the composite microbial inoculum is bacillus coagulans and bacillus amyloliquefaciens;
the bacillus coagulans is bacillus coagulans (Bacillus coagulans) MES847, and the preservation date is as follows: 8.30 days of 2018, the name of the preservation unit and short: china general microbiological culture collection center (CGMCC), number of collection: CGMCC No.16358;
the bacillus amyloliquefaciens is bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MES 812 and is preserved in China center for common microorganism strain preservation management, wherein the preservation address is North Celu No.1, no. 3 of the Korean area of Beijing city, the preservation date is 2017, 8 months and 10 days, and the preservation number is CGMCC No.14515;
the effective viable count of the bacillus amyloliquefaciens in the composite microbial inoculum is not less than 1.0x10 10 The effective viable count of bacillus coagulans per gram is not less than 5.0X10 8 Individual/g;
the mixed ratio of the bacillus coagulans bacterial powder and the bacillus amyloliquefaciens bacterial powder in the composite bacterial agent is 1:1.
2. The composite microbial agent for preventing and treating fish hepatobiliary syndrome according to claim 1, wherein bacillus coagulans and bacillus amyloliquefaciens are respectively cultured to prepare bacterial powder, and then mixed according to a proportion to prepare the composite microbial agent.
3. The preparation method of the compound microbial inoculum for preventing and treating fish hepatobiliary syndrome according to any one of claims 1-2, which is characterized by comprising the following steps: the bacillus coagulans and the bacillus amyloliquefaciens are subjected to shake flask culture, primary seed culture, secondary seed culture, fermentation, concentration and spray drying respectively to prepare bacterial powder, and then are mixed according to a proportion to prepare the composite bacterial agent.
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Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104411182A (en) * 2012-04-12 2015-03-11 杜邦营养生物科学有限公司 Microbial strains and their use in animals
CN103497906B (en) * 2013-08-02 2015-03-11 国家***第三海洋研究所 Microecological preparation, preparation method, and applications thereof
CN103966118B (en) * 2013-12-20 2015-11-18 中国水产科学研究院黄海水产研究所 Bacillus amyloliquefaciens and application thereof
CN105238722B (en) * 2015-11-03 2018-09-21 江苏省苏微微生物研究有限公司 The preparation method and application of one bacillus amyloliquefaciens bacterial strain and its bacterium powder preparation
CN108085265B (en) * 2016-11-22 2021-09-28 北京大北农科技集团股份有限公司 Bacillus coagulans new strain, and microecological preparation and feed thereof
WO2018155613A1 (en) * 2017-02-24 2018-08-30 東亜薬品工業株式会社 Composition for prevention and/or treatment of shrimp acute hepatopancreatic necrosis disease including bacillus amyloliquefaciens strain and/or treated product of said strain
CN107494999A (en) * 2017-09-20 2017-12-22 成都通威三新药业有限公司 A kind of the feature fish feed preparation and preparation method of liver protecting shield intestines
CN107616985A (en) * 2017-10-01 2018-01-23 青岛东海药业有限公司 Bacillus coagulans preparation and its application
CN108996711A (en) * 2018-09-11 2018-12-14 江苏远山生物技术有限公司 A kind of compound micro-ecological preparation used for aquiculture and preparation method thereof containing clostridium butyricum
CN111394275B (en) * 2020-02-22 2022-05-27 华东师范大学 Bacillus amyloliquefaciens and application thereof, aquatic feed and aquaculture method

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