CN112868942A - Macro-algae fermentation product capable of being used as bivalve shellfish bait and preparation method and application thereof - Google Patents
Macro-algae fermentation product capable of being used as bivalve shellfish bait and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a large-scale seaweed fermentation product capable of being used as a feed for bivalve shellfish and a preparation method and application thereof, belonging to the technical field of aquatic shellfish feeds. The large-scale seaweed fermentation product which can be used as the bait for the bivalve shellfish is obtained by fermenting large-scale seaweed and seawater shellfish intestinal microorganisms, has good palatability, can remarkably promote the growth of the bivalve shellfish, is suitable for being used as the bait for the bivalve shellfish, and fills the blank of the large-scale seaweed feed which is suitable for feeding shellfish seedlings at home.
Description
Technical Field
The invention belongs to the technical field of aquatic shellfish feeds, and particularly relates to a large-scale seaweed fermentation product capable of being used as bivalve shellfish bait, and a preparation method and application thereof.
Background
The shellfish industry is an important component of fishery in China, and shellfish culture and capture are important activities throughout coastal areas of China, and are leading production industries in some remote areas or even local areas. The shellfish industry which develops healthily plays an important or irreplaceable role in the aspects of guaranteeing national food safety, enriching animal protein types, maintaining the stability of coastal areas, widening employment channels, increasing income of fishermen, expanding export, earning foreign exchange, inheriting traditional culture, cleaning water quality in sea areas, fixing carbon and the like.
The shellfish mainly uses unicellular algae such as chrysophyceae, diatom, chlorella, etc. as bait. However, in the shellfish industrial artificial seedling breeding and pond intermediate cultivation processes, the problems of low culture yield, harsh culture conditions, high cost and the like of the unicellular algae often occur, and particularly, the unicellular algae is difficult to culture in the northern summer high-temperature period, so that the shellfish seedling bait is difficult to supply, the shellfish seedling survival rate is low, the yield is reduced, and the sustainable development of the shellfish industry is restricted.
The kelp contains carotenoid, protein, vitamin, P, iodine, Ca and the like, particularly polysaccharide unsaturated acid and the like which are nutrient components which are not contained in terrestrial plants and can meet the growth and development of marine shellfish. However, the large-sized seaweed has a large size of crushed particles and a large cell wall thickness, and thus it is difficult to use the large-sized seaweed as an artificial feed for efficient use of shellfish. At present, the research and development of marine algae feed suitable for shellfish seedling feeding at home still belong to a blank.
Disclosure of Invention
In order to solve the problems, the invention provides a large-sized seaweed fermentation product which can be used as a feed for bivalve shellfish and a preparation method and application thereof. The large-scale seaweed fermentation product provided by the invention can be used as bait for bivalve shellfish, and fills the blank of large-scale seaweed feed suitable for shellfish seedling ingestion in China.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a large-scale seaweed fermentation product which can be used as bait for bivalve shellfish, and the large-scale seaweed fermentation product is obtained by fermenting large-scale seaweed and seawater shellfish intestinal microorganisms.
Preferably, the macroalgae species include one or more of ulva, hemerocallis fulva, ulva pertusa, carrageen, gelidium amansii, sargassum thunbergii, kelp, wakame seaweed, sargassum muticum, sarcandra glabra, dactylophora, gloiophora, and sargassum fusiforme.
Preferably, the species of marine shellfish includes one or more of philippines, pacific oysters, and bay scallops.
Preferably, the fermentation time is 5-10 days, and the fermentation temperature is 20-25 ℃.
The invention provides a preparation method of a large-sized seaweed fermentation product in the technical scheme, which comprises the following steps: preparing a seawater shellfish intestinal microorganism suspension; mixing the large-sized seaweed ground liquid, the seawater shellfish intestinal microorganism suspension and seawater, and fermenting to obtain a large-sized seaweed fermentation product;
the viable count of microorganisms in the seawater shellfish intestinal microorganism suspension is not less than 1000 CFU/mL;
the volume ratio of the large-scale seaweed ground solution to the seawater shellfish intestinal microorganism suspension to the seawater is (80-120): (7.5-12.5): (80-120).
Preferably, the preparation method of the seawater shellfish intestinal microorganism suspension comprises the following steps: and (3) washing the seawater shellfish intestinal tract with sterile seawater, collecting the washed suspension, and homogenizing to obtain the seawater shellfish intestinal microorganism suspension.
Preferably, the predominant bacteria in the suspension of marine shellfish intestinal microorganisms include bacteria of phylum proteobacteria, phylum firmicutes, phylum bacteroides, phylum actinomycetes, cyanobacteria and phylum verrucomicrobia.
Preferably, the particle size of the kelp grinding fluid is 200-300 meshes.
Preferably, the method further comprises filtering the fermented liquid, wherein the filtering comprises rough filtering and fine filtering, the rough filtering is performed by a sieve with 10-20 meshes, and the fine filtering is performed by a sieve with 200-300 meshes.
The invention provides an application of the large-scale seaweed fermentation product in the technical scheme or the large-scale seaweed fermentation product obtained by the preparation method in the technical scheme in feeding bivalve shellfish offspring seeds, wherein the bivalve shellfish comprises Ruditapes philippinarum, mussels, clams, scallops and oysters.
Has the advantages that:
the invention provides a large-scale seaweed fermentation product which can be used as bait for bivalve shellfish, and the large-scale seaweed fermentation product is obtained by fermenting large-scale seaweed and seawater shellfish intestinal microorganisms. The large-scale seaweed fermentation product provided by the invention has good palatability, can obviously promote the growth of the bivalve shellfish, and can be used as bait of the bivalve shellfish. The results of the examples show that: the large-scale seaweed fermentation product is fed to Ruditapes philippinarum, the growth value of the shell length of the Ruditapes philippinarum is obviously higher than that of a feeding-free group and a feeding conventional feed group, and the large-scale seaweed fermentation product can be used as bait for bivalve shellfish.
Drawings
FIG. 1 is a graph showing the effect of each treatment of example 4 on clam shell length and shell height.
Detailed Description
The invention provides a large-scale seaweed fermentation product which can be used as bait for bivalve shellfish, and the large-scale seaweed fermentation product is obtained by fermenting large-scale seaweed and seawater shellfish intestinal microorganisms. In the present invention, the species of the macroalgae preferably include one or more of ulva, hemerocallis, ulva pertusa, carrageen, gelidium, sargassum thunbergii, kelp, undaria pinnatifida, sargassum pallidum, sarcandra, gloiopeltis, cuprum and sargassum fusiforme; more preferably one or more of ulva, hemerocallis, ulva pertusa and carrageen; more preferably ulva or hemerocallis; the invention selects the specific type of the large seaweed, so that the palatability of the fermentation product obtained by fermentation is better, and the growth of bivalve shellfish seedlings is promoted. In the present invention, the species of the seawater shellfish preferably include one or more of philippine clams, pacific oysters, and bay scallops; the invention selects specific species of seawater shellfish, the specific dominant phylum microorganism in the intestinal tract can effectively decompose large particles of crushed large algae into single-cell particles, and the large algae fermentation product obtained by fermenting the seawater shellfish intestinal microorganisms has good palatability, is beneficial to foraging of bivalve shellfish, solves the problem that the large algae is difficult to be used as artificial feed for shellfish high-efficiency utilization, and fills the blank of the domestic large algae feed suitable for shellfish seed ingestion. In the invention, the fermentation time is preferably 5-10 d, and more preferably 7 d; the fermentation temperature is preferably 20-25 ℃, and more preferably 25 ℃. The special fermentation conditions of the invention are very suitable for the growth of fermentation microorganisms, the efficiency of decomposing the large-sized seaweed into single cells is obviously improved, the fermentation process is simple, the fermentation can be completed within one week at room temperature, and the operability is strong.
According to the invention, the large-sized seaweed is fermented by shellfish intestinal microorganisms, so that the feed which has good palatability and can remarkably promote the growth of bivalve shellfish is provided, and the sustainable development of shellfish industry is promoted.
The invention provides a preparation method of a large-sized seaweed fermentation product in the technical scheme, which comprises the following steps: preparing a seawater shellfish intestinal microorganism suspension; mixing the large-sized seaweed ground liquid, the seawater shellfish intestinal microorganism suspension and seawater, and fermenting to obtain a large-sized seaweed fermentation product;
the viable count of microorganisms in the seawater shellfish intestinal microorganism suspension is not less than 1000 CFU/mL;
the volume ratio of the large-sized seaweed grinding liquid to the seawater shellfish intestinal microorganism suspension to the seawater is (80 mL-120): (7.5 mL-12.5): (80 mL-120).
The invention prepares seawater shellfish intestinal microorganism suspension. In the present invention, the preparation method of the suspension of intestinal microorganisms of seawater shellfish preferably comprises the following steps: and (3) flushing the inner side of the intestinal tract of the seawater shellfish with sterile seawater, collecting the flushed suspension, and homogenizing to obtain the suspension of the intestinal microorganisms of the seawater shellfish. The method of washing and the homogenization step are not particularly required in the present invention, and conventional methods are used by those skilled in the art. In the invention, the viable count of the microorganisms in the seawater shellfish intestinal microorganism suspension is not less than 1000CFU/mL, and more preferably 1000 CFU/mL. The preferred bacteria in the seawater shellfish intestinal microorganism suspension preferably comprise bacteria of proteobacteria, firmicutes, bacteroidetes, actinomycetes, cyanobacteria and verrucomicrobia. The invention provides rich strain resources for digesting the large-sized seaweed grinding fluid by utilizing various intestinal dominant bacteria, ensures that the large-sized seaweed is pre-digested before feeding the clams, and is convenient for the clams to be digested and utilized. The seawater shellfish intestinal microorganism suspension is mainly used for fermenting large-scale seaweed grinding liquid, so that large particles in the large-scale seaweed grinding liquid are decomposed to obtain single-cell particles, and the double-shell shellfish foraging is facilitated.
After obtaining the seawater shellfish intestinal microorganism suspension, the invention mixes the large-scale seaweed grinding liquid, the seawater shellfish intestinal microorganism suspension and seawater, and ferments to obtain the large-scale seaweed fermentation product. In the invention, the particle size of the large-sized seaweed grinding fluid is preferably 200-300 meshes; more preferably 300 mesh, and the fermentation efficiency can be improved by subjecting the macroalgae to a grinding treatment. The method for preparing the kelp grinding fluid has no special requirements, and the grinding method which is well known to the skilled person can be adopted. The invention mixes the large-sized seaweed grinding fluid, the seawater shellfish intestinal microorganism suspension and the seawater; the preferable volume ratio of the large-sized seaweed ground solution to the seawater shellfish intestinal microorganism suspension to the seawater is (80-120): (7.5-12.5): (80-120); more preferably 10: 1: 10. the invention can make the large-scale seaweed fully fermented by further selecting a specific fermentation proportion. The invention mixes the large-sized seaweed grinding fluid, the seawater shellfish intestinal microorganism suspension and the seawater and then carries out fermentation. In the invention, the fermentation time is preferably 5-10 d, and more preferably 7 d; the fermentation temperature is preferably 20-25 ℃, and more preferably 25 ℃.
After fermentation, the fermentation liquor after fermentation is preferably filtered to obtain the large seaweed fermentation product. In the invention, the filtration preferably comprises coarse filtration and fine filtration, wherein the coarse filtration is preferably sieved by a sieve with 10-20 meshes, and is further preferably sieved by a sieve with 20 meshes; the fine filtration is preferably carried out by a 200-300 mesh sieve, and is further preferably carried out by a 300 mesh sieve. The fermentation liquor after fine filtration is more suitable for filter-eating shellfish.
The preparation method of the large-sized seaweed fermentation product provided by the invention is simple to prepare, has wide raw material sources, and solves the problem that large-sized seaweed is difficult to be used as artificial feed for efficient utilization of shellfish.
The invention provides an application of the large-sized seaweed fermentation product in the technical scheme or the large-sized seaweed fermentation product obtained by the preparation method in the technical scheme in feeding bivalve offspring seeds. In the invention, the bivalve shellfish comprises Ruditapes philippinarum, mussels, clams, scallops and oysters, and preferably Ruditapes philippinarum. The large-sized seaweed fermentation product provided by the invention is used for feeding bivalve shellfish, can remarkably promote the growth of the length and the height of the bivalve shellfish, and is suitable for feeding bivalve shellfish seedlings.
In order to further illustrate the present invention, the following examples are given to describe the fermentation product of kelp which can be used as bait for bivalve shellfish and the preparation method and application thereof in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
A large-scale seaweed fermentation product which can be used as a bait for bivalve shellfish is prepared by the following steps:
1. selecting 10 healthy Ruditapes philippinarum, washing the shell with sterile seawater for 3 times, dissecting with a sterile scalpel to obtain an intestinal tract, washing the intestinal tract with sterile seawater for 3 times by using a sterile injector, collecting the washed suspension, homogenizing with a sterile homogenizer, and adding sterile seawater to 20mL to obtain the seawater shellfish intestinal tract microorganism suspension, wherein the viable bacteria number of microorganisms in the seawater shellfish intestinal tract microorganism suspension is not lower than 1000 CFU/mL.
2. Weighing 100g of fresh ulva lactuca, washing with sterilized pure water, and grinding in a sterilized grinding mill to obtain about 100mL of large-scale seaweed ground liquid with the particle size of about 300 meshes.
3. Selecting a 500mL conical flask, cleaning, and putting into an oven for drying for later use.
4. Putting the conical flask into the prepared large-scale seaweed ground solution, and adding 10mL of seawater shellfish intestinal microorganism suspension and 100mL of sterilized seawater.
5. Putting the conical flask into a constant-temperature incubator, adjusting the temperature to 25 ℃, and fermenting for 7 d.
6. And (3) roughly filtering the fermentation liquor by using a 20-mesh silk screen, and then filtering by using a 300-mesh silk screen to obtain ulva fermentation liquor, wherein the filtered product is the large-scale seaweed fermentation product which can be used as the bait for the bivalve shellfish.
Example 2
A large-scale seaweed fermentation product which can be used as a bait for bivalve shellfish is prepared by the following steps:
1. selecting 10 healthy Ruditapes philippinarum, washing the shell with sterile seawater for 3 times, dissecting with a sterile scalpel to obtain an intestinal tract, washing the intestinal tract with sterile seawater for 3 times by using a sterile injector, collecting the washed suspension, homogenizing with a sterile homogenizer, and adding sterile seawater to 20mL to obtain the seawater shellfish intestinal tract microorganism suspension, wherein the viable bacteria number of microorganisms in the seawater shellfish intestinal tract microorganism suspension is not lower than 1000 CFU/mL.
2. Weighing 100g of fresh hemerocallis fulva, washing with sterilized pure water, and grinding in a sterilized grinding mill to obtain about 100mL of large-sized seaweed ground liquid with particle size of 300 mesh.
3. Selecting a 500mL conical flask, cleaning, and putting into an oven for drying for later use.
4. Putting the conical flask into the prepared large-scale seaweed ground solution, and adding 10mL of seawater shellfish intestinal microorganism suspension and 100mL of sterilized seawater.
5. Putting the conical flask into a constant-temperature incubator, adjusting the temperature to 25 ℃, and fermenting for 7 d.
6. And (3) roughly filtering the fermentation liquor by using a 20-mesh silk screen, and then filtering by using a 300-mesh silk screen to obtain the hemerocallis fermentation liquor, wherein the filtered product is the macroalgae fermentation product which can be used as the bait for the bivalve shellfish.
Example 3
The methods of example 1 and example 2 were used to prepare a large-sized seaweed fermentation product, each method was used to prepare two batches continuously, total DNA was extracted from the obtained large-sized seaweed fermentation product and from samples of intestinal tissue of ruditapes philippinarum used in the test, and the microbial community structure of each sample was examined (stored at-80 ℃ and examined by bemaimer), and the examination results are shown in tables 1 and 2.
TABLE 1 Ruditapes philippinarum intestinal bacterial flora composition
The composition of the healthy philippine clam intestinal bacterial community for fermenting the large-scale seaweed ground liquid is researched through 16S rDNA high-throughput sequencing, and the clam intestinal bacterial community is mainly characterized by Proteobacteria, Firmicutes, Bacteroides, Actinobacilla, Cyanobacteria and Verrucomicrobia. Although the structures of intestinal bacterial communities of different clam individuals are different, the dominance gates are basically consistent, and the final fermentation product cannot be influenced by the difference among different clam individuals.
TABLE 2 Macro-algal fermentation broth bacterial community composition
The bacterial community composition of the fermentation liquid of the two large-scale seaweeds including the hemerocallis fulva and the ulva is researched through 16S rDNA high-throughput sequencing, and the bacterial community dominant phyla of the fermentation liquid of different large-scale seaweeds and different batches of the fermentation liquid include Proteobacteria (Proteobacteria), Firmicutes (Firmicutes), EpsilonBacaeota, Cyanobacteria (Cyanobacter), Fusobacteria (Fusobacteria), bacteroides (bacteriodes), Verrucomicrobia (Verrucomicrobia) and the like, are basically the same as the dominant phyla of the intestinal tract of the philippines, and the difference between different clam individuals does not influence the final fermentation product.
Example 4
The macroalgae fermentation liquid of example 1 and example 2 was fed to Ruditapes philippinarum, and a blank control group, a conventional bait feeding group, and a non-fermented macroalgae ground liquid feeding group were set. Wherein, the conventional feed group is fed with chrysophyceae and Chaetoceros every day, and the daily feeding amount is 5000 cells/mL (the final concentration of chrysophyceae and Chaetoceros in the barrel after feeding); the blank control group was not fed; the daily feeding amount of the other treatment groups was 5000 cells/mL (cells were counted in regular and irregular shapes in the fermentation broth of macroalgae; large numbers of regular cells were still visible in the ground macroalgae liquid, and counted in regular cells). The shell length and shell height increase values of the clams of different test groups after three weeks of feeding are recorded, and the detection results are shown in table 3 and figure 1; SL in FIG. 1 is the shell length; SH is shell height; the control is no feeding; mock is to feed conventional baits (golden algae, Chaetoceros); HMF is a fermentation liquor fed with hemerocallis fulva; SCF is feeding ulva fermentation liquor; HMF + SCF is a fermentation liquid for feeding hemerocallis fulva and an ulva fermentation liquid; HM is the fresh milled solution fed with non-fermented hemerocallis fulva (i.e. the fresh milled solution fed with non-fermented hemerocallis fulva in table 3); SC feeding non-fermented ulva fresh grinding liquid (namely feeding non-fermented ulva grinding liquid in the table 3); HM + SC is the feeding of the fresh milling solution of non-fermented hemerocallis fulva and the fresh milling solution of non-fermented ulva (i.e. the feeding of the fresh grinding solution of non-fermented hemerocallis fulva and the fresh milling solution of non-fermented ulva in Table 3).
TABLE 3 influence of different feeding modes on the clam shell length and shell height
As can be seen from the results of Table 3 and FIG. 1, the growth value of the clam shell length was the highest in the case of the ulva fermentation product fed alone, and was 349. + -. 53. mu.m; the growth value of the shell of young clam is 315 +/-69 mu m when the young clam is fed with the hemerocallis fulva fermentation product alone. The shell length growth values for the clams in both test groups were significantly higher than the shell length growth values for the clams in the no-feed group and the conventional bait group (p < 0.05). The shell length and shell height increasing values of the young clams fed with the fresh grinding fluid test group of the non-fermented hemerocallis, the fresh grinding fluid test group of the non-fermented ulva, the fresh grinding fluid test group of the non-fermented hemerocallis and the fresh grinding fluid test group of the non-fermented ulva are obviously lower than the shell length and shell height increasing values of the young clams fed with the conventional bait (p is less than 0.05). The results in table 3 and figure 1 suggest that the fermentation product obtained after milling macroalgae and co-fermenting with healthy philippine clam gut microbes can be used as a bait for clams.
The results of the above examples show that the kelp fermentation product provided by the invention has good palatability, can significantly promote the growth of bivalve shellfish, and can be used as bait for bivalve shellfish; meanwhile, the preparation method of the large-scale seaweed fermentation product provided by the invention is simple, the raw material source is wide, and the blank of the large-scale seaweed feed suitable for shellfish offspring seed ingestion in China is filled.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (10)
1. A large-scale seaweed fermentation product capable of being used as a bait for bivalve shellfish is characterized in that the large-scale seaweed fermentation product is obtained by fermenting large-scale seaweed and seawater shellfish intestinal microorganisms.
2. The macroalgae fermentation product of claim 1, wherein the macroalgae species comprise one or more of ulva lactuca, hemerocallis fulva, ulva pertusa, carrageen, gelidium amansii, sargassum thunbergii, kelp, wakame seaweed, sargassum muticum, sarcandra glabra, gloiopeltis canadensis, cuprum algae, and hizikia fusiforme.
3. The kelp fermentation product according to claim 1, wherein the species of marine shellfish comprises one or more of philippines, pacific oysters, and gulf scallops.
4. The kelp fermentation product according to claim 1, wherein the fermentation time is 5-10 days and the fermentation temperature is 20-25 ℃.
5. A method for preparing a kelp fermentation product according to any one of claims 1 to 4, comprising the steps of: preparing a seawater shellfish intestinal microorganism suspension; mixing the large-scale seaweed ground solution, the seawater shellfish intestinal microorganism suspension and seawater, and fermenting to obtain a large-scale seaweed fermentation product;
the viable count of microorganisms in the seawater shellfish intestinal microorganism suspension is not less than 1000 CFU/mL;
the volume ratio of the large-scale seaweed ground solution to the seawater shellfish intestinal microorganism suspension to the seawater is (80-120): (7.5-12.5): (80-120).
6. The method for preparing the suspension of the marine shellfish intestinal microorganisms as claimed in claim 5, wherein the method for preparing the suspension of the marine shellfish intestinal microorganisms comprises the following steps: and (3) washing the seawater shellfish intestinal tract with sterile seawater, collecting the washed suspension, and homogenizing to obtain the seawater shellfish intestinal tract microorganism suspension.
7. The method of claim 5, wherein the predominant bacteria in the suspension of marine shellfish intestinal microorganisms comprises bacteria of the phylum Proteobacteria, Thelephorata, Bacteroides, Actinomycetes, cyanobacteria, and verrucomicrobia.
8. The method according to claim 5, wherein the kelp grinding fluid has a particle size of 200 to 300 mesh.
9. The preparation method according to claim 5, further comprising filtering the fermented liquid, wherein the filtering comprises coarse filtering and fine filtering, the coarse filtering is performed by a 10-20 mesh sieve, and the fine filtering is performed by a 200-300 mesh sieve.
10. The application of the large-scale seaweed fermentation product of any one of claims 1 to 5 or the large-scale seaweed fermentation product obtained by the preparation method of any one of claims 6 to 9 in feeding bivalve shellfish offspring seeds, wherein the bivalve shellfish comprises Ruditapes philippinarum, mussels, clams, scallops and oysters.
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| 洪奕娜等: "藻菌混合固定化胶球在海水贝类育苗中的应用初探", 《安徽农学通报(上半月刊)》 * |
| 邹建威: "岩扇贝内脏团和肠道可培养微生物及其宏基因多样性分析", 《中国优秀博硕士学位论文全文数据库（硕士） 农业科技辑》 * |
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