CN105285514A - Plant lactobacillus feed capable of relieving tilapia lead toxicity - Google Patents
Plant lactobacillus feed capable of relieving tilapia lead toxicity Download PDFInfo
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- CN105285514A CN105285514A CN201510823741.4A CN201510823741A CN105285514A CN 105285514 A CN105285514 A CN 105285514A CN 201510823741 A CN201510823741 A CN 201510823741A CN 105285514 A CN105285514 A CN 105285514A
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
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Abstract
The invention discloses a plant lactobacillus feed capable of relieving tilapia lead toxicity, and belongs to the technical field of feed. The feed is a finished feed probiotics granulated feed having the viable bacterial number of more than 10<8> cfu/g and prepared by a way of adding a bacterial agent obtained by carrying out centrifuging concentration or freeze drying and microcapsule embedding on CGMCC 5494 strains into an aquatic animal feed pre-mix material in a form of a feed additive, and then granulating or pre-granulating and then spraying probiotics on a carrier to obtain the feed. The feed obtained by the method can increase the growth rate of tilapia, increases the intestinal tract digestive enzyme activity of the lead-exposure tilapia, reduces the lead content in bodies of the lead-exposure tilapia, relieves the oxidative stress damage in the bodies of the lead-exposure tilapia, improves the activity of aminolevtllinic acid dehydratase in the bodies of the lead-exposure tilapia, improves the lysozyme activity and myeloperoxidase activity of the lead-exposure tilapia, relieves the cell and genetic toxicity of the lead-exposure tilapia, and better has application prospects.
Description
Technical field
The present invention relates to a kind of Lactobacillus plantarum feed can alleviating Tilapia mossambica Lead Toxicity, belong to field of feed.
Background technology
Lead is the very common Heavy Metallic Elements of occurring in nature, has very strong toxicity to human body.The World Health Organization in 1993 and FAO (Food and Agriculture Organization of the United Nation) (FAO/WHO) combine human body that the code committee recommends, and can to tolerate Lead intake be 25 μ g/ (kg body weight week).Metallic lead quality is soft, and be one of metal of applying the earliest of the mankind, its main application makes lead accumulator, is widely used in RONA, building, metal working material, munitions, trichromoscope and radiation shield etc.Because lead-acid accumulator provides energy to preserve, dual-use productive life all be unable to do without it.Along with the progress forward of economic society, especially industrialized acceleration is advanced, and the problem of lead contamination becomes increasingly conspicuous, and the lead contamination of water environment is also more and more severeer.There is report, enter over 10 years, along with the industrialization that China advances always, the industry of touching lead contamination also gets more and more, comprise heavy metal exploitation, metal smelt, chemical industry, printing and dyeing, the energy etc., also have the outstanding problem such as indiscriminate exploitation, illegal blowdown of some violation enterprises, the generation of lead contamination accident is also more and more frequent.Due to the characteristic of lead contamination, a large amount of lead enters aquaculture system, causes murder by poisoning to aquatile, and produce acute, chronic disease, the safety in utilization of aquatic products is threatened, has had a strong impact on the ecological balance of fish production and water environment, and then human health is worked the mischief.
Lead has carried out decades to the toxicologic study of animal, and research shows that lead poisoning can cause damage in various degree to each system in body.Heavy metal lead enters after in aquatic organism, accumulation can be caused, passive impact is caused on its health simultaneously, almost poison organs all in health, affect normal physiological function, and have potential teratogenesis, mutagenesis harm, form chronic or acute poisoning symptom, the death rate of aquatic life time serious, may be caused to rise.Current research finds that lead poisoning is mainly reflected in the following aspects to hydrobiological toxic action: A, affects immune system; B, infringement hemopoietic system; C, affects internal system; D, infringement reproductive system.
The saturnine Therapy study of aquatic animal is less, and its main thought is Adsorption of Lead or alleviates the plumbous toxic action caused.Current research mainly adds certain antioxidant or plumbous adsorbent to hydrobiological Treatment of Lead Poisoning method in feed.Antioxidant mainly some vitamins and polyphenoils comprises vitamin A, vitamin C, vitamin E, thiamine and mineral matter selenium, involved by the research of plumbous adsorbent has on imvite, zeolite and attapulgite.And antioxidant use cost is higher, adsorbent uses the ionic equilibrium may destroyed in aquatic organism.
In view of the finiteness of current present Research and methods for the treatment of, seem very necessary for hydrobiological lead accumulation and a kind of new intervention of lead poisoning exploration or methods for the treatment of; And the feasibility of above-mentioned several methods for the treatment of also impels us to continue, and searching is applied more extensively, potentiality are huger, there are again the means alleviating Lead Toxicity effect simultaneously.Probio is extensively present in human intestine, spontaneous fermentation dairy products and vegetalitas fermented food as among pickles, sauerkraut.Long-Term Scientific Study result shows, probio is that human body is requisite and have the beneficial bacterium of important physiological function, and its main Physiological Function has: control lactose intolerance, recover colony balance in human body intestinal canal and safeguard health, antitumor and pre-anti-cancer effect, control the functions such as toxins in human body level.Have researcher verified, the lactic acid bacteria of some specific bacterial strain can effective Adsorption of Heavy Metal Ions, and has significant effect at alleviation oxidative stress and oxidative damage.Lactic acid bacteria as microorganism have cost low, without environmental hazard, can be used as the features such as feed addictive.And, there is a large amount of scholar's research probiotic feed to the prebiotic effect of aquatic livestock, but do not had people to report in the lead poisoning of alleviation aquatile.Therefore explore one and can alleviate the saturnine probiotic feed of aquatic animal, its practice significance is huge.Therefore a kind of lactobacillus feed alleviating Lead Toxicity of exploratory development just further can excavate the function of probio, develops the probio be possessed of higher values, and alleviates aquatic animal Lead Toxicity open up the approach and solution that make new advances for utilizing probiotic additive.
Summary of the invention
The object of this invention is to provide a kind of Lactobacillus plantarum feed; described Lactobacillus plantarum feed carries out after carrying out in premix Lactobacillus plantarum CGMCCNo.5494 being joined aquatic animal feed with the form of feed addictive granulating or granulate in advance preparing probiotic feed containing viable bacteria in the mode of carrier spraying probio, is greater than 10 in described Lactobacillus plantarum feed containing viable bacteria content
8the Lactobacillus plantarum CGMCCNo.5494 of CFU/g.
Described Lactobacillus plantarum CGMCCNo.5494 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 29th, 2011, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, and deposit number is CGMCCNo.5494.
Involved Lactobacillus plantarum CGMCCNo.5494 has following biological characteristics:
(1) thalline feature: Gram-positive, cell is rod-short, and thalline is about 0.5-1.0 μm wide, and 2-4 μm is long, Cheng Dan, paired or chaining, and do not form gemma, two ends are circular;
(2) colony characteristics: form obvious bacterium colony on MRS culture medium, diameter is between 0.3-2.0mm, circular, milky, and translucent, surface wettability is smooth, neat in edge, not chromogenesis;
(3) growth characteristics: the minimum growth temperature of this bacterial strain is 20 DEG C, and maximum growth temperature is 40 DEG C, at temperature 30-37 DEG C, growth is best, and the highest tolerant and minimum initial growth pH is 9.0 and 2.5, and the initial pH of the most suitable growth is 6.0; The period of delay of lactobacillus plantarum strain of the present invention is relatively short, and about 4h starts to enter exponential phase, and 12h just reaches stationary phase.
Involved Lactobacillus plantarum CGMCCNo.5494 joins in aquatic animal feed as feed addictive, also has following beneficial characteristics:
(1) growth rate of Tilapia mossambica can be improved;
(2) the plumbous enteron aisle digestive enzyme activity exposing Tilapia mossambica can be improved;
(3) lead content, the especially lead content of Tilapia mossambica liver pancreas, kidney, spleen and the gill in plumbous exposure Tilapia mossambica body can be reduced;
(4) plumbous exposure Tilapia mossambica vivo oxidation stress damage can be alleviated;
(5) the plumbous activity exposing aminolevulinate dehydratase (ALAD) in Tilapia mossambica body can be improved;
(6) the plumbous lysozyme activity and the activity of myeloperoxidase (MPO) that expose Tilapia mossambica can be improved;
(7) the plumbous cell and the genetoxic that expose Tilapia mossambica can be alleviated.
In one embodiment of the invention, described feed addictive is the concentrated bacterium liquid of CGMCCNo.5494 or the freeze-dried vaccine powder of freeze drying process acquisition, or the sodium alginate micro gel capsule microbial inoculum of CGMCCNo.5494.
Described carrier is acceptable carrier in feed.
Described in feed acceptable carrier be one or more normally used adhesive, lubricant or protective agents in feed.
In one embodiment of the invention, the preparation of described concentrated bacterium liquid comprises the following steps:
A. the preparation of culture medium: use and 10% enzyme hydrolysis skimmed milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract are dissolved in the water by culture medium gross mass 87.7%, then adjusting its pH is 6.8, obtains culture medium;
B. Lactobacillus plantarum CGMCC5494 bacterial classification is inoculated in the culture medium at temperature 110-120 DEG C after sterilizing 8-12min according in the inoculum concentration of the quality 2-4% of described culture medium, then at temperature 37 DEG C, 18h is cultivated, with pH7.2 phosphate buffer cleaning 2-4 time, reach 10 by the resuspended viable bacteria concentration that makes of physiological saline
11cFU/mL, obtains fresh concentrated bacterium liquid.
In one embodiment of the invention, the preparation of described freeze-dried vaccine powder comprises the following steps:
C. the preparation of freezing drying protective agent: use water and the standby protective agent obtained containing 100g/L skimmed milk power, 30mL/L glycerine, 100g/L maltodextrin, 150g/L trehalose, 10g/LL-sodium glutamate of protective agent mixed raw material;
D. 10 are reached according to way phosphate buffer cleaning 2-4 rear resuspended viable bacteria concentration that makes of freezing drying protective agent with step C preparation of steps A, B
11cFU/mL, allows this suspension preculture 60min at temperature 37 DEG C, then carries out freeze drying and obtain described freeze-dried vaccine powder.
In one embodiment of the invention, the preparation of described sodium alginate micro gel capsule microbial inoculum comprises following step:
E. the viable bacteria concentration of CGMCC5494 is reached 10
11the concentrated bacterium liquid of more than CFU/mL, be that 1:10 fully mixes with 2% sodium alginate soln with volume ratio, and add the Tween 80 of 0.4%, magnetic stirrer emulsification 15min, use drive nozzle to spray in the CaCl2 solution of 0.2mol/L, solidification 15min, by filtered through gauze, normal saline flushing three times, obtains sodium alginate micro gel capsule microbial inoculum.
In one embodiment of the invention, described Tilapia mossambica basal feed formula and trophic component are as shown in following table 1, table 2:
Table 1 basal feed composition and content
Note:
apremix provides for every kilogram of feed: vitamin A, 3000IU; Vitamin D, 1500IU; Vitamin E, 150mg; Vitamin K, 15mg; Cobastab
1, 10mg; Cobastab
2, 20mg; Cobastab
6, 10mg; Cobastab
12, 0.1mg; Vitamin C, 500mg; Calcium pantothenate, 20mg; Nicotinic acid, 25mg; Folic acid, 5mg; Inositol, 150mg; Choline, 2000mg; Biotin, 1mg; Calcium, 4g; Magnesium, 600mg; Iron, 60mg; Copper, 6mg; Manganese, 50mg; Zinc, 100mg; Selenium, 0.2mg; Iodine, 1mg; Cobalt, 1mg.
Table 2 basal feed composition and nutrient composition content
Feed making method described in one embodiment of the present invention as shown in Figure 1.
The application of Lactobacillus plantarum CGMCCNo.5494 of the present invention in fish meal can accelerate the growth rate of Tilapia mossambica, improve enteron aisle digestive enzyme activity, reduce the lead content of the internal organs such as liver pancreas, kidney, the gill in Tilapia mossambica body, the oxidativestress damage that the lead poisoning of alleviation Tilapia mossambica body internal cause causes, the reduction of aminolevulinate dehydratase activity, immune level decline and cell and genetoxic, very have application prospect.
Accompanying drawing explanation
Fig. 1 is feed making method flow;
Fig. 2 is feed CGMCCNo.5494 activity (a) of different addition manner and the Changing Pattern of survival rate (b) at 4 DEG C;
Fig. 3 is feed CGMCCNo.5494 activity (a) of different addition manner and the Changing Pattern of survival rate (b) at 25 DEG C;
Fig. 4 is Tilapia mossambica enteron aisle amylase (A) and intestinal protease (B) activity change; The group indicating different letter represents significant difference (P<0.05);
Fig. 5 is the MDA change of Tilapia mossambica liver pancreas (A) and trunk kidney (B); The group indicating different letter represents significant difference (P<0.05);
Fig. 6 is the reductive glutathione change of Tilapia mossambica liver pancreas (A) and trunk kidney (B); The group indicating different letter represents significant difference (P<0.05);
Fig. 7 is the glutathione peroxidase change of Tilapia mossambica liver pancreas (A) and trunk kidney (B); The group indicating different letter represents significant difference (P<0.05);
Fig. 8 is the TAC change of Tilapia mossambica liver pancreas (A) and trunk kidney (B); The group indicating different letter represents significant difference (P<0.05);
Fig. 9 is aminolevulinate dehydratase activity change in Tilapia mossambica serum; The group indicating different letter represents significant difference (P<0.05);
Figure 10 is that Tilapia mossambica lysozyme (A) is active with myeloperoxidase (B); The group indicating different letter represents significant difference (P<0.05);
Figure 11 is Tilapia mossambica erythrocyte nucleus form: A-B, normal cell core; C-D, kidney shape core; E-F, splits core; G-H, emits complex; I-J, cavity core; K-L, hook-shaped core; M-N, double leaf core; O-P, micronucleus; In figure, scale is 4 μm.。
Detailed description of the invention
Embodiment 1 mixes tests the impact of different probio addition manner feed CGMCCNO.5494 activity
The concentrated bacterium liquid made by CGMCCNO.5494, freeze-dried vaccine powder, sodium alginate micro gel capsule microbial inoculum add in Tilapia mossambica basal feed respectively, mash feed is mixed into by mixer, with bacterium colony colony counting method, the viable bacteria content in mash feed is detected, acquired results is multiplied by feed relative, is designated as M; To being added into the concentrated bacterium liquid before feed, freeze-dried vaccine powder and sodium alginate micro gel capsule microbial inoculum carry out bacterium colony plate count, calculate total viable count, are designated as M0, obtain survival rate=M/M0 × 100%.Result is as shown in table 3:
Table 3 mixes the impact on different probio addition manner feed CGMCCNO.5494 survival rate
Note: each group does not all have significant difference
Known by table 3, there is no the significance difference opposite sex (p>0.05) between each group.Illustrate that mixed cell operation does not affect substantially on the activity of CGMCCNO.5494.
Embodiment 2 is granulated and to be tested the impact of different probio addition manner feed CGMCCNO.5494 activity
The concentrated bacterium liquid made by CGMCCNO.5494, freeze-dried vaccine powder, sodium alginate micro gel capsule microbial inoculum add in Tilapia mossambica basal feed respectively, are mixed into mash feed by mixer, adopt bacterium colony plate count to detect to the viable bacteria content in mash feed; Granulated in granulator by mixed mash feed, pelleting temperature is 70 DEG C, granulates and terminates to carry out the detection of bacterium colony plate count to the viable bacteria content in pellet afterwards.Result is as shown in table 4:
Table 4 is granulated on the impact of different probio addition manner feed CGMCC5494 survival rate
Note: different letter represents between group has significant difference (P<0.05)
Pelletization unit operation is larger to different addition manner CGMCCNO.5494 activity influence.Affect comparatively large on concentrated bacterium liquid addition manner and freeze-dried vaccine powder addition manner, its survival rate has been reduced to 6% and 2.54% respectively.After adopting sodium alginate to carry out microcapsules process, its survival rate significantly promotes, and reaches 24.01%, improves 4 times and 8 times respectively compared to two other group.Illustrate that sodium alginate micro gel capsule can effectively protect CGMCCNO.5494 of the present invention to survive under severe processing environment.
Embodiment 3 is stored and is tested the impact of different probio addition manner feed CGMCCNO.5494 activity
Store two months under being put in 4 DEG C and 25 DEG C of conditions respectively with the pellet of concentrated bacterium liquid, freeze-dried vaccine powder, sodium alginate micro gel capsule and rear spray process of granulating, week about a bacterium colony plate count is carried out to pellet and detect viable count and calculate its survival rate.
Experimental result is as shown in accompanying drawing 2, accompanying drawing 3, can find out under 4 DEG C of conditions by accompanying drawing 2, the CGMCCNO.5494 vigor of each group all shows good stability, as shown in accompanying drawing 2 (a), within one month, the CGMCCNO.5494 activity of four groups all can remain on the same order of magnitude.In the store experiment of 2 months, sodium alginate micro gel capsule group shows best stability, and the 8th week time, its viable count still can remain on about 40%.Shown in accompanying drawing 2 (b), when the 7th week, concentrated bacterium liquid interpolation group starts to occur reducing largely, and alginic acid difficult microcapsules interpolation group survival rate is the highest, is 44.61%.When the 8th week, what decline degree was maximum is freeze-dried vaccine powder interpolation group, and the survival rate of its viable bacteria drops to 3.38%.Secondly after granulating, spraying probiotic group, be 8.16%, and sodium alginate micro gel capsule group survival rate still can remain on 40.77%.Under 25 DEG C of conditions, nearly all group all shows inadaptability, as shown in accompanying drawing 3 (a), when the 5th week, all can't detect viable count.Except sodium alginate micro gel capsule group, other groups, the 3rd week time, can't detect viable count.The viable count of sodium alginate micro gel capsule group within the previous moon will be significantly higher than other three groups (P<0.05).After granulating, the stability of spraying probiotic group is also poor, and as shown in accompanying drawing 3 (b), the 2nd Zhou Shiqi can't detect viable count.This illustrates that 4 DEG C have the activity being beneficial to protection CGMCCNO.5494; and sodium alginate micro gel capsule interpolation CGMCCNO.5494 can keep more stable activity in 4 DEG C; slow down the decrease speed of CGMCC5NO.494 at 25 DEG C of condition vigor, the activity of protection CGMCC5494.
Embodiment 4 Lactobacillus plantarum CGMCCNO.5494 exposes the impact experiment of Growth Op Tilapia and digestive enzyme activity to lead
Choose healthy Tilapia mossambica 180 tail, original body mass mean+SD is 53.40 ± 1.25g, is divided into four groups at random, often organizes 15 tails, in triplicate.Divide into groups respectively, control group: basal diet of feeding; Pb exposure group: basal diet of feeding, is exposed to the Pb of 1mg/L
2+in solution; CGMCC5494 intervention group: CGMCC5494 probiotic feed of feeding, is exposed to the Pb of 1mg/L
2+in solution; Probio CGMCC5494 group: CGMCC5494 probiotic feed of feeding.The CGMCC5494 feed used in embodiment 2 with the pellet that concentrated bacterium liquid mode makes.Test 28 days by a definite date, period the record scale of feeding of Tilapia mossambica and death condition, record Tilapia mossambica body weight after terminating, puts to death Tilapia mossambica and dissects, and gets enteron aisle and builds up kit according to Nanjing respectively and national standard prolease activity assay method measures enteron aisle amylase and prolease activity.
These experimental results are as shown in table 5 and accompanying drawing 4.As shown in Table 5, contrast CGMCC5494 group and control group, Pb exposure group and CGMCC5494 intervention group, find all have significant difference on average weight gain and daily gain, illustrates that probio CGMCC5494 facilitates the plumbous growth exposing Tilapia mossambica.
The growth indexes change of table 5 Tilapia mossambica
Note: between the group of different letter representative, there is significant difference (P<0.05)
From accompanying drawing 4, add two groups (CGMCC5494 intervention group and probio CGMCC5494 group) of probio CGMCC5494, the trend of digestive enzyme activity is higher than contrast and Pb exposure group.The amylase activity of CGMCC5494 intervention group and prolease activity improve about 100%, 100% than Pb exposure group respectively, 40%, 33% is improve respectively than probio CGMCC5494 group, illustrate that CGMCC5494 can significantly improve the plumbous exposure enteron aisle endo-amylase of Tilapia mossambica and the vigor of protease, facilitate the plumbous growth exposing Tilapia mossambica.
In addition, the present invention also in the same way, has investigated the impact of Lactobacillus plantarum LP2-3 on Growth Op Tilapia and digestive enzyme activity.Similar in CGMCC5494, Lactobacillus plantarum LP2-3 has the adsorption effect of good lead ion equally in lead ion solution, result but shows and exposes the facilitation of growth of Tilapia mossambica significantly lower than the present invention to lead, and cannot be improved significantly to the exposure enteron aisle endo-amylase of Tilapia mossambica and the vigor of protease.
Embodiment 5 Lactobacillus plantarum CGMCC5494 exposes the reducing effect experiment of lead content in Tilapia mossambica body to lead
Zoopery and grouping is carried out with Tilapia mossambica according to described in embodiment 4, experiment terminates rear execution Tilapia mossambica and dissects, get liver pancreas, kidney, spleen, the gill, brain, muscle of back and gonadal tissue, put into Hyperfrequency waves eliminating stove and clear up 20min, the digestion solution obtained uses atomic absorption spectrometer (SpectrAA220, Varian, USA) measure lead content.Experimental result is as shown in table 6.
Table 6 Tilapia mossambica internal organs lead content
Note :-expression does not detect; Unit is μ g/g tissue wet; The group indicating different letter represents significant difference (P<0.05).
Contrast Pb+5494 group and Pb group can find, in liver pancreas, the gill, brain, sexual gland, dorsal muscles etc. are organized, both present significant difference, and in trunk kidney and spleen, otherness is not quite, but its trend remain probio intervention group will lower than lead exposure group.These data illustrate that CGMCC5494 can reduce the plumbous lead content exposed in Tilapia mossambica body.
Embodiment 6 Lactobacillus plantarum CGMCC5494 is alleviated the plumbous Tilapia mossambica oxidative damage that exposes and is tested
Zoopery and grouping is carried out with Tilapia mossambica with described in embodiment 4, experiment terminates rear execution Tilapia mossambica and dissects, get liver pancreas and kidney, the method building up reagent test box according to Nanjing measures the level of MDA (MDA), reductive glutathione (GSH), glutathione peroxidase (GSH-Px), TAC (T-AOC) respectively.Test result as depicted in figures 5-8.
As depicted in figures 5-8, CGMCC5494 alleviates because lead exposes the reduction of GSH, GSH-Px and the T-AOC caused, and the rising of MDA in liver.GSH is the important substance maintaining cell biological function, and GSH-Px is a kind of antioxidase extensively existed in body, poisonous peroxide catalyzed can be become innocuous substance, the biomarker of both heavy metal exposures; T-AOC is the overall target for weighing oxidation resistance, and its size can reflect the state of body autoxidation metabolism, under lead exposes, can produce oxidative damage, cause T-AOC to reduce; And MDA is the end-product of free radical in a kind of organism and lipid generation peroxidization, is a kind of marker enzyme of organism internal oxidition stress damage.These results suggest that, CGMCC5494 alleviates the oxidative stress and oxidative damage that Tilapia mossambica lead poisoning causes.
Embodiment 7 Lactobacillus plantarum CGMCC5494 is alleviated aminolevulinate dehydratase (ALAD) activity in plumbous exposure Tilapia mossambica body and is reduced experiment
Carry out zoopery and grouping with described in embodiment 4 with Tilapia mossambica, experiment terminates rear execution Tilapia mossambica and dissects, and tail bone vein gets Tilapia mossambica blood, and measure according to the kit of Chuan Xiang bio tech ltd, Shanghai, measurement result as shown in Figure 9.
ALAD is one of key enzyme of the former synthesis of ferroheme, and directly relevant with hemopoietic system, lead poisoning can suppress several key enzymes of the former synthesis of ferroheme, comprises ALAD, is saturnine action target spot enzyme, and the activity of ALAD can as saturnine biomarker.Found by contrast, plumbous exposure group content comparatively other each groups significantly declines, and illustrate that plumbous exposure inhibits ALAD active, and Pb+5494 significantly improves ALAD activity, illustrates that CGMCC5494 can protect ALAD active in saturnine damage.
Embodiment 8 Lactobacillus plantarum CGMCC5494 improves immune level experiment in plumbous exposure Tilapia mossambica body
Zoopery and grouping is carried out with Tilapia mossambica with described in embodiment 4, experiment terminates rear execution Tilapia mossambica and dissects, tail bone vein gets Tilapia mossambica blood, and build up kit according to Nanjing and carry out mensuration lysozyme and myeloperoxidase (MPO) activity, measurement result as shown in Figure 10.
Lysozyme and MPO are parts for nospecific immunity, and lysozyme can be engulfed and kill inoculating microbe, and MPO also can kill the microorganism engulfed in cell.This two indices represents the power of nospecific immunity to a certain extent.From accompanying drawing 10, due to the impact that lead exposes, the lysozyme content of plumbous group and the active mean value of MPO are all lower than control group, and Pb+5494 group lysozyme content and MPO activity are significantly higher than Pb group; Illustrate that CGMCC5494 significantly improves lead and exposes the lysozyme content and the reduction of MPO activity that cause.
Embodiment 9 Lactobacillus plantarum CGMCC5494 alleviates plumbous exposure Tilapia mossambica cell and Genetic toxicity
Zoopery and grouping is carried out with Tilapia mossambica with described in embodiment 4, experiment terminates rear execution Tilapia mossambica and dissects, tail bone vein is got Tilapia mossambica blood and is made blood smear, examine under a microscope, amplify 1000 times, random number 1000 red blood cells, the statistics heterokaryotic number of different classification (accompanying drawing 11).Result is as shown in table 7.
Table 7 poikilocytosis nuclear statistics
Note: unit is individual/1000 red blood cells; The group indicating different letter represents significant difference (P<0.05)
Table 7 lists the heterokaryotic statistic of classification of red born of the same parents.Can reflect the toxicity of environmental contaminants to genotoxic inspection, and micronucleus and core abnormity are to hereditary and that cytotoxicity is very responsive index.As shown in Table 7, plumbous group at red blood cell kidney shape core, split number on core double leaf core, micronucleus and be significantly higher than blank group, and kidney shape core to be heteromorphic nucleus occur that quantity is maximum, it can be said that the bright reason exposed due to lead creates genetoxic and cytotoxicity to Tilapia mossambica.Contrast Pb group and Pb+5494 group, find the heteromorphic nucleus number of Pb+5494 group at kidney shape core, split core, emit little complex, remarkable in Pb group in the frequency of occurrences of cavity core, hook-shaped core, double leaf core and micronucleus, illustrate that CGMCC5494 reduces heteromorphic nucleus number, the heredity that the lead poisoning can alleviating Tilapia mossambica causes and the cytotoxicity under plumbous exposure as probiotic additive.
Embodiment 10 utilizes Lactobacillus plantarum CGMCC5494 to make the pellet containing this bacterium
Lactobacillus plantarum CGMCC5494 is cultivated 18h in the medium, fresh concentrated bacterium liquid, freeze-dried vaccine powder and sodium alginate micro gel capsule microbial inoculum is obtained according to the method addressed before, be added into respectively in powdery premixing feedstuff and fully mix, granulate with minitype fodder-pelleting machine, pelleting temperature is 70 DEG C, obtains the pellet containing this bacterium; After pellet is shaping, uses drive nozzle to be evenly mixed in pellet fresh concentrated bacterium liquid, obtain the pellet containing this bacterium.The viable count of gained pellet is greater than 10
8cFU/g finished product feedstuff.
Although the present invention with preferred embodiment openly as above; but it is also not used to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; all can do various changes and modification, what therefore protection scope of the present invention should define with claims is as the criterion.
Claims (10)
1. a Lactobacillus plantarum feed; it is characterized in that; described Lactobacillus plantarum feed carries out after carrying out in premix Lactobacillus plantarum CGMCCNo.5494 being joined aquatic animal feed with the form of feed addictive granulating or granulate in advance preparing probiotic feed containing viable bacteria in the mode of carrier spraying probio, is greater than 10 in described Lactobacillus plantarum feed containing viable bacteria content
8the Lactobacillus plantarum CGMCCNo.5494 of CFU/g.
2. Lactobacillus plantarum feed according to claim 1, it is characterized in that, described aquatic animal feed is Tilapia mossambica basal feed, according to the mass fraction, containing Soybean Meal 40%, rice bran 14%, flour 22%, fish meal 10%, Cottonseed Meal 5%, rapeseed dregs 5%, soybean oil 2%, premix 2%.
3. Lactobacillus plantarum feed according to claim 2, is characterized in that, described feed addictive is the microbial inoculum that CGMCCNo.5494 obtains with centrifugal concentrating or freeze drying process, or the sodium alginate micro gel capsule microbial inoculum of CGMCCNo.5494.
4. Lactobacillus plantarum feed according to claim 1, is characterized in that, described carrier is acceptable carrier in feed.
5. method according to claim 4, is characterized in that, described in feed acceptable carrier be one or more normally used adhesive, lubricant or protective agents in feed.
6. the arbitrary described Lactobacillus plantarum feed of claim 1-5 is improving the application in the plumbous enteron aisle digestive enzyme activity exposing Tilapia mossambica.
7. the arbitrary described Lactobacillus plantarum feed of claim 1-5 is improving the plumbous application exposing the growth rate of Tilapia mossambica.
8. the arbitrary described Lactobacillus plantarum feed of claim 1-5 improve plumbous expose aminolevulinate dehydratase in Tilapia mossambica body active in application.
9. the application of the arbitrary described Lactobacillus plantarum feed of claim 1-5 in the lysozyme activity improving plumbous exposure Tilapia mossambica and activity of myeloperoxidase.
10. a feed addictive, is characterized in that, described feed addictive is the microbial inoculum utilizing Lactobacillus plantarum CGMCCNo.5494 to prepare.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105724788A (en) * | 2016-03-30 | 2016-07-06 | 马家庆 | Method for manufacturing breeding feed of eumeces chinensis in pre-egg stage |
| CN105961916A (en) * | 2016-05-13 | 2016-09-28 | 中山大学 | Probiotics-containing tilapia feed |
| CN106616042A (en) * | 2016-12-23 | 2017-05-10 | 江南大学 | Lactobacillus plantarum feed and application thereof |
| CN106666081A (en) * | 2017-01-13 | 2017-05-17 | 江南大学 | Probiotics feed capable of relieving aluminium toxicity of tilapia and application thereof |
| CN107594136A (en) * | 2017-09-04 | 2018-01-19 | 安徽秀安生态农业有限公司 | A kind of sorbefacient doll's fish meal |
| CN107668389A (en) * | 2017-09-04 | 2018-02-09 | 安徽秀安生态农业有限公司 | A kind of doll's fish meal |
| CN111040960A (en) * | 2019-11-19 | 2020-04-21 | 西南大学 | Lactobacillus plantarum LP33 and application thereof in preparation of product for promoting lead excretion |
| CN111197012A (en) * | 2018-11-17 | 2020-05-26 | 原生生物医学股份有限公司 | Novel lactobacillus plantarum, lactobacillus composition and use thereof for treating or preventing heavy metal-related diseases |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586148A (en) * | 2012-02-28 | 2012-07-18 | 江南大学 | Plant lactobacillus capable of relieving lead toxicity and application thereof |
-
2015
- 2015-11-24 CN CN201510823741.4A patent/CN105285514A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586148A (en) * | 2012-02-28 | 2012-07-18 | 江南大学 | Plant lactobacillus capable of relieving lead toxicity and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| 王汗成: "植物乳杆菌饲料缓解铅对罗非鱼的毒性作用", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
| 王汗成等: "含益生菌罗非鱼颗粒饲料的加工方式及贮存稳定性研究", 《安徽农业科学》 * |
| 田丰伟: "缓解氧化应激乳酸菌的筛选、表征和功能评价研究", 《中国博士学位论文全文数据库工程科技1辑》 * |
| 高鹏飞等: "乳酸菌在水产养殖业中的应用", 《家畜生态学报》 * |
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| CN105724788A (en) * | 2016-03-30 | 2016-07-06 | 马家庆 | Method for manufacturing breeding feed of eumeces chinensis in pre-egg stage |
| CN105961916A (en) * | 2016-05-13 | 2016-09-28 | 中山大学 | Probiotics-containing tilapia feed |
| CN106616042A (en) * | 2016-12-23 | 2017-05-10 | 江南大学 | Lactobacillus plantarum feed and application thereof |
| CN106666081A (en) * | 2017-01-13 | 2017-05-17 | 江南大学 | Probiotics feed capable of relieving aluminium toxicity of tilapia and application thereof |
| CN107594136A (en) * | 2017-09-04 | 2018-01-19 | 安徽秀安生态农业有限公司 | A kind of sorbefacient doll's fish meal |
| CN107668389A (en) * | 2017-09-04 | 2018-02-09 | 安徽秀安生态农业有限公司 | A kind of doll's fish meal |
| CN111197012A (en) * | 2018-11-17 | 2020-05-26 | 原生生物医学股份有限公司 | Novel lactobacillus plantarum, lactobacillus composition and use thereof for treating or preventing heavy metal-related diseases |
| CN111040960A (en) * | 2019-11-19 | 2020-04-21 | 西南大学 | Lactobacillus plantarum LP33 and application thereof in preparation of product for promoting lead excretion |
| CN111040960B (en) * | 2019-11-19 | 2022-02-01 | 西南大学 | Lactobacillus plantarum LP33 and application thereof in preparation of product for promoting lead excretion |
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