CN105060499B - A kind of compound micro-ecological preparation for improving breeding water body transparency and its application - Google Patents

Abstract

The invention provides a kind of compound micro-ecological preparation for improving breeding water body transparency and its application, the probiotics is to be formed by different strains with carrier compounding, and wherein bacillus amyloliquefaciens bacterial content is not less than 5 × 10⁹ CFU/g, simple bacillus bacterial content are not less than 1 × 10¹⁰ CFU/g, Lactobacillus plantarum content are not less than 2 × 10⁹CFU/g.The compound micro-ecological preparation has stronger capacity of decomposition, can effectively decompose the organic matters such as residual bait, excrement in pond, significantly improves water transparency, and stable water body pH, regulation algae is balanced each other, purified water.

Description

A kind of compound micro-ecological preparation for improving breeding water body transparency and its application

Technical field

The invention belongs to micro ecological technology field, it is related to a kind of compound micro-ecological preparation, and in particular to one kind improves cultivation The compound micro-ecological preparation of water transparency and its application.

Background technology

" breed fish and first support water ", water colour and transparency are the important indicators of pond culture.Transparency can influence whole water body Stability and water fertility, stabilization for whole water body, fishes and shrimps are grown up healthy and sound, the preventing and treating of disease has positive work With.With the increase of aquaculture density, the increase of daily ration, feeding quantity, metabolite, the organic matter such as excrement, residual bait can bulk deposition, water There is the relatively low deterioration water quality of the transparencies such as muddy water, yellow mud Ba Shui, Heisui River in cognition.

Microorganism can increase dissolved oxygen, reduction ammonia nitrogen and nitrite nitrogen as a kind of ecological regulator, suppress harmful micro- Biological breeding and rapid degradation of organic substances, maintenance aquatic ecological balance.The purifying water micrcxDrganism main species applied at present have light Close bacterium, bacillus, nitrobacteria, saccharomycete etc..In highdensity aquaculture process, bacillus etc. is added in good time Microorganism formulation etc., can accelerate ammonia nitrogen, nitrite etc. in the material circulation of water body, reduction breeding water body, commonly using helping Keep stable in water body, both less " fertilizer " was not easy to become " clear " water, and water environment is in good level for a long time.

The content of the invention

For degradation of organic substances, reduction ammonia nitrogen, improve breeding water body environment, breeding water body is improved the invention provides one kind The compound micro-ecological preparation of transparency and its application, the present invention can significantly carry two kinds of bacillus and lactic acid bacteria compounding use High water transparency, purifies water.

For achieving the above object, the technical scheme is that：

A kind of compound micro-ecological preparation for improving breeding water body transparency, the compound micro-ecological preparation includes bacterial content not Less than 5 × 10⁹CFU/g bacillus amyloliquefaciens, bacterial content is not less than 1 × 10¹⁰CFU/g simple bacillus and bacterium contain Amount is not less than 2 × 10⁹CFU/g Lactobacillus plantarum and carrier.

The simple bacillus is from deposit number：CGMCC No. 10051 simple bacillus f5.

The bacillus amyloliquefaciens are from deposit number：CGMCC No. 10011 bacillus amyloliquefaciens HFJ- 7。

Described carrier is talcum powder, and 60 mesh oversizes are 0.

The invention provides application of the described compound micro-ecological preparation in breeding water body transparency is improved.

The compound micro-ecological preparation is watered full pool spilling head, and consumption is 200~250g/ mus, is splashed once within 10~15 days.

The breeding water body includes the breeding water body of prawn, grass carp, silver carp, carp and loach.

Compared with prior art, the present invention has advantages below and technique effect：Simple bacillus that the present invention is selected, Organic matter, ammonia nitrogen, the nitrous acid of bacillus amyloliquefaciens, Lactobacillus plantarum to cultivating the later stage etc. has significant degradation effect, Organic oxygen demand is reduced, indirect oxygenation maintains algae phase, bacterium to balance each other, and reduces water body pH, and purification improves water quality, promotes cultivation dynamic Thing well grows.Complex microorganism preparations of the present invention are put on into water body, the transparency of breeding water body can be improved, had Extensive market application foreground.

Brief description of the drawings

Fig. 1 is influence of the compound micro-ecological preparation to water transparency；

Fig. 2 is influence of the compound micro-ecological preparation to water body ammonia nitrogen.

Embodiment

The present invention combines example in detail below, and the present invention is further detailed explanation.

The screening of embodiment 1, bacterial strain

First, simple bacillus f5 separation screening

In July, 2014, water sample, bed mud from the coastal many places collection prawn culturing region in Jiangnan are that sample screens Bacillus. Bed mud takes supernatant to be sample by conventional doubling dilution；Water sample can be directly as sample；Sample liquid is incubated in after 75 DEG C of 20min, It is coated on nutrient agar panel and carries out the separation of bacterial strain, purifying, obtains 4 bacillus.

Purified single strain carries out ammonia nitrogen degradation measure.Single strain is inoculated in LB seed culture mediums, 37 DEG C 180 Rpm cultivate about 20h to cell concentration be 10⁸ CFU/mL.By 1% inoculum concentration（Volume ratio）It is forwarded in ammonia nitrogen culture medium（Glucose 5g, MgSO₄·7H₂O 0.25g, K₂HPO₄0.5g, (NH₄)₂SO₄7.2~7.4,20 DEG C of 150rpm trainings of 0.1g, water 1L, pH Support）, timing sampling measure ammonia nitrogen concentration, the bacterium（Numbering f5）To ammonia nitrogen degradation effect preferably, degradation rate is 99% during 36h.

Through Physiology and biochemistry method Preliminary Identification, bacterial strain f5 is bacillus, its biological characteristics：It is Gram-positive, good Oxygen rod bacterium, 15 DEG C~40 DEG C of growth temperature.Bacterium colony is circular, and neat in edge, milky, bacterium colony is smooth, relatively moistens.

Using bacterial strain f5 DNA as template, 16S rRNA universal primers are that primer is expanded, and amplified fragments carry out sequence survey It is fixed.Sequencing result is compared with the 16S rRNA sequences of Blast softwares category kind related to GenBank, shows bacterial strain 16S In rRNA sequences and GenBank gene pools in bacillusBacillus simlpe16s rRNA sequence homologies degree most Height, homology reaches 99%.The 16S rRNA sequences of existing bacillus in Genbank are lost by DNAMAN6.0 Pass evolutionary analysis, as a result show, bacterial strain f5 16S rRNA withBacillus simpleHomology highest, judges the bacterial strain as bud The simple bacillus of spore Bacillus.

The bacterial strain f5 screened is subjected to culture presevation, depositary institution：China Committee for Culture Collection of Microorganisms is general Logical microorganism center（CGMCC）；Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica； Preservation date：On November 25th, 2014；Simple bacillusBacillus simplexDeposit number be CGMCC No.10051。

2nd, bacillus amyloliquefaciens HFJ-7 separation screening

In January, 2012 in March, 2012, from 10, the town suitable for reading in Shouguang City of Weifang City of Shandong Province, capsicum green house of vegetables takes 30 Capsicum Rhizosphere sampling sheet.Soil sample is fully ground respectively, in heating 20min in 75 DEG C of water-baths, then uses sterile saline In 1000 times of dilutions, in dilution spread on nutrient agar NA flat boards, the line purifying of picking single bacterium colony filters out 68 plants of pure bacterium altogether, It is bacillus through microscopy, including bacterial strain HFJ-7.

Through Bacterial Physiological biochemical method Preliminary Identification, bacterial strain HFJ-7 is bacillus, its biological characteristics：Gram Stained positive, the colony diameter formed on nutrient agar panel about 3-3.5mm, edge is irregular, and middle fold is faint yellow. Thalli morphology is in shaft-like, gemma ellipse, and end life, size is 0.84 × 0.52 μm.

Using molecular genetics methods to the further taxonomic identifications of bacterial strain bacillus HFJ-7.Bacterial strain HFJ-7 is extracted first DNA, using the DNA as template, 16S rRNA universal primers be primer, 16S rRNA fragments are expanded, amplified fragments enter Row sequencing.Sequencing result is compared with the 16S rRNA sequences of Blast softwares category kind related to GenBank, as a result Show bacterial strain 16S rRNA sequences and the 16s of the Bacillus pumilus in bacillus in GenBank gene pools RRNA sequence homology degree highests, homology reaches 99%.By DNAMAN6.0 to existing bacillus in Genbank 16S rRNA sequences carry out phylogenetic analysis, as a result show, bacterial strain S3 16S rRNA withBacillus amyloliquefaciensHomology highest, judges bacillus amyloliquefaciens of the bacterial strain as bacillus.

The bacillus amyloliquefaciens HFJ-7 screened is subjected to culture presevation, depositary institution：Chinese microorganism strain preservation Administration committee's common micro-organisms center（CGMCC）；Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences Institute of microbiology；Preservation date：On November 19th, 2014；Bacillus amyloliquefaciensBacillus amyloliquefaciensDeposit number be CGMCC No. 10011.

Embodiment 2, the culture of simple bacillus

F5 single bacterium colonies are inoculated in LB seed culture mediums, 37 DEG C of 180rpm culture about 20h to cell concentration are 10⁸ CFU/mL；By 1% inoculum concentration（Volume ratio）Fermentation medium is forwarded to, the fermentative medium formula is：Corn steep liquor 1g, grape Sugar 3 g, yeast extract 0.5 g, KH₂PO₄0.1 g, K₂HPO₄0.15 g, (NH₄)₂SO₄0.5 g, NaC1 0.1 g, MgSO₄· 7H₂O 0.05 g, H₂O 100 mL, pH 7.0~7.2.

37 DEG C of 180rpm cultivate 24h, microscopy gemma maturing rate>90%；Counted using nutrient agar panel, bacterium amount>150 ×10⁸ CFU/mL。

Embodiment 3, the application in leg Shrimp Litopenaeus vannamei Culture Raceway-type Ponds

The compound micro-ecological preparation of the present invention for improving breeding water body transparency contains bacterial content and is not less than 5 × 10⁹ CFU/g bacillus amyloliquefaciens, bacterial content is not less than 1 × 10¹⁰CFU/g simple bacillus, bacterial content be not less than 2 × 10⁹CFU/g Lactobacillus plantarum and carrier.The simple bacillus is from deposit number：CGMCC No.'s 10051 Simple bacillus f5, the bacillus amyloliquefaciens are from deposit number：CGMCC No. 10011 solution starch gemma bar Bacterium HFJ-7, the Lactobacillus plantarum uses commercially available strain, and carrier is talcum powder, and 60 mesh oversizes are 0.

The more muddy prawn Tu Tang of 44 mu or so of selection, water quality is research object.The fine day morning is according to 1 mu of 200g microbial inoculum It is watered full pond uniformly to splash, splashes and aerator is opened after finishing 2 hours, continuous tracing detection index 7 days, the water of timing observation daily Color change, determines the indexs such as water transparency, ammonia nitrogen, nitrous acid.Water transparency is measured using black and white disk method.For guarantee test The accuracy of progress, without products such as other chemical classes, biological species, the disinfectant classes of influence result of the test during experiment.

As a result as shown in figure 1, the transparency of 1~4# pond waters increases with observing time, gradually tend towards stability.It is logical The field observation to water body and microscope initial survey are crossed, discovery is splashed after compound micro-ecological preparation, and planktonic microalgae quantity is equally corresponding It increased.From figure 2 it can be seen that bacillus has significant degradation effect to ammonia nitrogen, 4 pond ammonia nitrogen degradation amounts are equal >50%, anaphase effect remains stable.During experiment, probiotics of splashing influences little to water body pH, each pond pH change Change amplitude maintains 0.2~0.3, pH and is kept between 8.2~8.4.

Embodiment 4, the application in grass carp cultivates

Test site is splashed compound micro-ecological preparation by 1 mu of 200g consumption, sprinkled within every 10 days in Hubei grass carp plant Spill once, while determining the indexs such as ammonia nitrogen, nitrous acid, pH, the transparency of cultivating pool water body.In the whole breeding process of control group Without using probiotics.

As a result show, using the probiotics water body environment and breeding production performance can be made to obtain excellent to a certain extent Change, the death rate of reduction cultivation grass carp significantly reduces ammonia nitrogen, content of nitrite, respectively 72.5%, 55.9% (P<0.05)； Water transparency averagely improves 31.2%, pH and is stably maintained between 8.2~8.6.

The compound micro-ecological preparation is watered full pool spilling head, and consumption is 200~250g/ mus, is splashed once within 10~15 days；Water Body is muddy serious optionally continuously to splash 3~5 days.

The above embodiments are merely illustrative of the technical solutions of the present invention, rather than is limited；Although with reference to foregoing reality Apply example the present invention is described in detail, for the person of ordinary skill of the art, can still implement to foregoing Technical scheme described in example is modified, or carries out equivalent substitution to which part technical characteristic；And these are changed or replaced Change, the essence of appropriate technical solution is departed from the spirit and scope of claimed technical solution of the invention.

Claims (4)

1. a kind of compound micro-ecological preparation for improving breeding water body transparency, it is characterised in that：The compound micro-ecological preparation by Following components is constituted：Bacterial content is not less than 5 × 10⁹CFU/g bacillus amyloliquefaciens, bacterial content is not less than 1 × 10¹⁰ CFU/ G simple bacillus and bacterial content are not less than 2 × 10⁹CFU/g Lactobacillus plantarum and carrier；The simple bacillus It is from deposit number：CGMCC No. 10051 simple bacillus f5；

The bacillus amyloliquefaciens are from deposit number：CGMCC No. 10011 bacillus amyloliquefaciens HFJ-7.

2. the compound micro-ecological preparation according to claim 1 for improving breeding water body transparency, it is characterised in that：Described Carrier is talcum powder, and 60 mesh oversizes are 0.

3. application of the compound micro-ecological preparation in breeding water body transparency is improved described in claim 1, it is characterised in that will The compound micro-ecological preparation is watered full pool spilling head, and consumption is 200~250g/ mus, is splashed once within 10~15 days.

4. application of the compound micro-ecological preparation according to claim 3 in breeding water body transparency is improved, its feature exists In the breeding water body is to include the breeding water body of prawn, grass carp, silver carp, carp and loach.