CN103865839B - One kind of probiotics applied to aquaculture - Google Patents

One kind of probiotics applied to aquaculture Download PDF

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CN103865839B
CN103865839B CN 201410014890 CN201410014890A CN103865839B CN 103865839 B CN103865839 B CN 103865839B CN 201410014890 CN201410014890 CN 201410014890 CN 201410014890 A CN201410014890 A CN 201410014890A CN 103865839 B CN103865839 B CN 103865839B
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CN103865839A (en )
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崔云龙
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青岛东海药业有限公司
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Abstract

本发明涉及一种应用于水产养殖的制剂,具体是由花域芽孢杆菌制成的制剂在水产养殖中预防疾病、提高饲料转化率或净化养殖水质中的应用。 The present invention relates to a formulation for used in aquaculture, in particular a preparation made by flower fields Bacillus prophylaxis of diseases in aquaculture, feed conversion or purification applications of Water Quality.

Description

一种应用于水产养殖的微生态制剂 One kind of probiotics applied to aquaculture

技术领域 FIELD

[0001] 本发明涉及一种应用于水产养殖的制剂,具体是由花域芽孢杆菌制成的制剂在水产养殖中预防疾病、提高饲料转化率或净化养殖水质中的应用。 [0001] The present invention relates to a formulation for used in aquaculture, in particular made of a formulation flower domain Bacillus preventing a disease in aquaculture, feed conversion or the water quality purifying breeding applications.

背景技术 Background technique

[0002] 2006年底在全国爆发的大菱鲆药残事件给我们敲响了警钟。 [0002] in the country by the end of 2006 outbreak of turbot Yaocan event to us sounded the alarm. 欧盟己于2006年全面禁止抗菌药作为饲料添加剂在养殖中的应用。 The EU had in 2006 a total ban on antibiotics as feed additives applied in the breeding of. 为保证食品安全,美国等发达国家对肉类、海产品等抗菌药残留进行检测。 To ensure food safety, the United States and other developed countries for meat, seafood and other antibacterial drugs residues were detected. 随着中国加入世贸组织,名优产品出口被普遍看好。 With China's accession to the WTO, famous export is generally good. 但由于国内养殖不科学使用抗生素等有害药物的现象严重,大大限制了产品的出口。 However, due to the domestic aquaculture unscientific use of antibiotics and other harmful drugs is serious, greatly limiting exports. 开发抗菌药替代产品应用于养殖业是目前市场所需要的。 Development of alternative antibiotics used in aquaculture is required by the market.

发明内容 SUMMARY

[0003] 本发明的目的是提供一种在水产养殖中预防疾病、提高饲料转化率或净化养殖水质的制剂,该制剂由花域芽孢杆菌制成。 [0003] The object of the present invention is to provide a method for preventing diseases in aquaculture, feed conversion or the water quality purifying breeding formulation, the formulation is made by a Bacillus flower field.

[0004] 本发明的制剂的制备优选是通过下述步骤实施的,但不局限于此制备工艺,公知的能实现的制备工艺均可以:采取含花域芽孢杆菌的样本,然后将样品置于灭菌瓶内,4°C 保存,研究时从中取一定量样品加入18mL灭菌的稀释液中,充分混匀,在无菌操作台内,进行10—\10—2、10—3、10—4,10-5,10_6,10_7梯度稀释,取10—5,10'10-7三个稀释梯度,分别涂布于花域芽孢杆菌选择性单菌落分离固体培养基上,置于培养箱内,30°C下培养48小时,选择长势良好的单菌落分别接种于花域芽孢杆菌液体扩增培养基中,3(TC下扩增培养48小时。将所得培养液离心(12000rpm)分离出菌体后,将菌体冷冻真空干燥,调制出各菌种的干燥菌粉,然后根据《伯杰细菌鉴定手册》、相关细菌鉴定文献或16S rRNA序列对比分析进行花域芽孢杆菌菌种鉴定和毒性试验,将无毒性的花域芽 [0004] The preparation of the formulations of the present invention is preferably implemented by the following steps, but is not limited to this preparation process, known production process can be realized can be: taking a sample containing Bacillus flower field, the samples were then placed sterilized bottle, 4 ° C storage, a certain amount from the study of the sample diluent is added 18mL sterilized, thoroughly mixed, in a sterile console, a 10- \ 10-2,10-3,10 -4,10-5,10_6,10_7 dilutions, diluted take 10-5,10'10-7 three gradient fields are applied to flowers Bacillus selectively isolated single colony on solid medium, placed in an incubator under the culture, 30 ° C 48 hours, the selection of single colonies growing well were inoculated Bacillus flower field amplification liquid medium, and cultured for 48 hours amplification (at 3 TC. (12000rpm) the resulting culture was centrifuged out after the bacterial cells, the cells were freeze-dried in vacuo, to prepare a dried bacterial powder of each species, and domain analysis of spent Bacillus species identified in accordance with "Manual bacterial identification Berger," bacterial identification related documents, or 16S rRNA sequence comparison and toxicity tests, the non-toxic flower buds domain 杆菌干燥菌粉按需要比例添加辅料制成颗粒剂、片剂、胶囊、散剂、粉剂或液体制剂等各种剂型,或制成药品、兽药、饲料添加剂或水质净化剂,也可加入其他活菌或低聚果糖等低聚糖起到协同作用。 Bacillus dried bacterial powder added in the desired ratio auxiliary materials in various dosage forms of granules, tablets, capsules, powders, powders, or liquid formulations and the like, or made into pharmaceuticals, veterinary medicines, feed additives or water purification agents, can also be added to other viable oligosaccharides or fructooligosaccharides synergistic effect.

[0005] 花域芽孢杆菌选择性单菌落分离固体培养基优选但不局限于:纯化水1L,蛋白胨l〇_〇g,牛肉浸膏3.(^,恥(:15.(^,琼脂25.(^,调?117.2,121。(:,灭菌15分钟。 [0005] Flower domain single colony isolated Bacillus selective solid medium is preferably but not limited to: purified water 1L, l〇_〇g peptone, beef extract 3 (^, shame (: 15 (^, agar 25 . (^ tune? 117.2,121. (:, sterilized for 15 minutes.

[0006] 花域芽孢杆菌液体扩增培养基优选但不局限于:纯化水1L,蛋白胨5.0g,酵母粉1 • 〇g,柠檬酸铁0 • lg,氯化钠I9 • 45g,氯化镁5.98g,硫酸钠3.24g,氯化钙1.8g,氯化钾0 • 55g,碳酸钠0.16g,溴化钾0 • 08g,氯化锶0 • 〇34g,硼酸0.022g,硅酸钠0 • 004g,氟化钠0.0024g,硝酸钠0 • 0016g,磷酸氢二0 • 008g,调pH7 • 4,121。 [0006] Bacillus flower field amplification liquid medium preferably but not limited to: purified water 1L, peptone 5.0g, yeast extract 1 • 〇g, ferric citrate 0 • lg, sodium chloride I9 • 45g, Magnesium chloride 5.98g , 3.24 g of sodium sulfate, calcium chloride 1.8g, KCl 0 • 55g, sodium carbonate 0.16g, KBr 0 • 08g, 0 • 〇34g strontium chloride, 0.022 g boric acid, sodium silicate 0 • 004g, 0.0024 g sodium fluoride, sodium nitrate 0 • 0016g, dicalcium phosphate 0 • 008g, adjusting pH7 • 4,121. (:,灭菌15分钟。 (:, Sterilized for 15 minutes.

[0007]为进一步具体说明本发明,发明人利用上述方法通过花域芽孢杆菌选择性单菌落分离固体培养基分离鉴定出了无毒性的花域芽孢杆菌,本发明所述花域芽孢杆菌并不局限于为说明本发明所述的花域芽孢杆菌,只要无毒性的花域芽孢杆菌均是本发明所述,均在本发明的保护范围内。 [0007] In order to further illustrate the present invention, the inventors took domain by the above method by Bacillus selectively isolated single colony isolated and identified a solid medium, non-toxic domain of Bacillus flowers, the flower of the present invention is not domain Bacillus to illustrate the limited domain of Bacillus flower according to the present invention, as long as the non-toxic domain of Bacillus flowers are the present invention are within the scope of the present invention.

[0008]本发明所述花域芽孢杆菌优选但不限于花域芽孢杆菌DHH28保藏编号CGMCC8561, 花域芽孢杆菌DHH06保藏编号CGMCC8566。 [0008] The present invention preferably take domain but not limited to Bacillus subtilis flower field DHH28 Accession No. CGMCC8561, flower fields Bacillus DHH06 Accession No. CGMCC8566.

[0009]本发明实施说明中优选使用的花域芽孢杆菌的细菌学性质,本发明并不局限本发明所述细菌学性质,只是为说明本发明的优选: [0009] The embodiment of the present invention will be described bacteriological properties of Bacillus flower domain preferably used, the present invention is not limited to the bacteriological properties of the present invention, only to illustrate the present invention preferably:

[0010] 1、为说明本发明,本发明利用上述方法分离的花域芽孢杆菌分别为花域芽孢杆菌DHH28保藏编号CGMCC8561,花域芽孢杆菌DHH06保藏编号CGMCCS566。 Separation [0010] 1, to illustrate the present invention, the present invention is a method of using the flowers are flowers domain Bacillus subtilis domain DHH28 Accession No. CGMCC8561, flower fields Bacillus DHH06 Accession No. CGMCCS566.

[0011] 2、菌体和菌落形态 [0011] 2, and cell colony morphology

[0012]显微镜观察:长2.0-2.5微米,宽0.7-0.9微米,长杆状,革兰氏染色阳性,芽孢呈椭圆形中生。 [0012] microscope: 2.0-2.5 m long, 0.7 to 0.9 microns wide, a long rod-shaped, gram positive, spore oval in green.

[0013] 菌落形态呈近圆形,边缘整齐且粘稠湿润,白色不透明。 [0013] colony morphology were nearly circular, neat edge and viscous moist, white opaque.

[0014] 3、生理生化鉴定 [0014] 3, physiological and biochemical identification

[0015] 硫化氢:一;卩引味:一;明胶液化:+ ;硝酸盐还原:一。 [0015] Hydrogen sulfide: a; Jie primer taste: a; Liquefaction of gelatin: +; Reduction of nitrate: a.

[0016] 4、糖酵解实验鉴定 [0016] 4, experimental identification glycolysis

[0017] 葡萄糖:+。 [0017] Glucose: +.

[0018] 本发明所述的花域芽孢杆菌指活生物个体。 [0018] Flower domain according to the present invention refers to a living organism Bacillus individual.

[0019]本发明是以有效剂量的上述花域芽孢杆菌作为主要活性成份,按照一定的制剂工艺,加入常规的赋形剂、调味剂、崩解剂、防腐剂、润滑剂、湿润剂、黏合剂、溶剂、增稠剂、增溶剂等辅料,制成任何一种适合于使用的剂型,如片剂、胶囊剂、颗粒剂、散剂、液体制剂、粉剂等剂型。 [0019] The present invention is based on the effective dose of Bacillus flower domain as a main active ingredient, according to certain preparation process, the addition of conventional excipients, flavoring agents, disintegrants, preservatives, lubricants, wetting agents, bonding agents, solvents, thickeners, solubilizing agents and other accessories, made suitable for use any one of forms, such as tablets, capsules, granules, powders, liquids, powders and other formulations.

[0020] 本发明所述以有效剂量的上述花域芽孢杆菌作为主要活性成份制成活菌制剂,也可以是上述花域芽孢杆菌与双歧杆菌、链球菌、枯草芽孢杆菌等益生菌中的一种或几种或其他活性成份组合制成活菌制剂,以起到协同作用,提高效果。 [0020] In the present invention, an effective dose of the above-described flower domain of Bacillus as a main active ingredient preparation made viable, and may be spent above Bacillus domain and Bifidobacterium, Streptococcus, Bacillus subtilis probiotics in one or more other active ingredients or combination thereof viable agents to synergistic effect, improving effect.

[0021] 本发明所指有效剂量是指以上述花域芽孢杆菌根据上面所述单独或组合作为主要药物活性成份制成的固体活菌制剂包含的总活菌数不能低于1 x 10sCFU/g,一般在1 X 109CFU/g以上,最高可达到1 X 10nCFU/g或1 X 10nCFU/g以上。 [0021] The present invention is referred to an effective dose refers to the total number of viable cells viable solid formulation according to the above-described flower Bacillus domain alone or in combination as described above is made mainly of a pharmaceutical active ingredient comprises not less than 1 x 10sCFU / g , typically 1 X 109CFU / g or more, up to 1 X 10nCFU / g or 1 X 10nCFU / g or more.

[0022] 本发明所指有效剂量是指以上述花域芽孢杆菌根据上面所述单独或组合作为主要药物活性成份制成的液体活菌制剂包含的总活菌数不能低于1 X 10sCFU/mL,一般在1 X 109CFU/mL 以上,最高可达到1 X 10nCFU/mL或1 X 10uCFU/mL 以上。 [0022] The present invention is referred to an effective dose refers to the total number of viable cells viable liquid formulation according to the above-described flower Bacillus domain alone or in combination as described above is made mainly of a pharmaceutical active ingredient comprises not less than 1 X 10sCFU / mL , typically 1 X 109CFU / mL or more, up to 1 X 10nCFU / mL or 1 X 10uCFU / mL or more.

[0023] 本发明首次公开花域芽孢杆菌在水产养殖中预防疾病、提高饲料转化率或净化养殖水质中的应用,因此含上述花域芽孢杆菌的制剂在上述中的应用均属于本发明的保护范围。 [0023] First Publication flower domain Bacillus of the present invention is the prevention of diseases in aquaculture, feed conversion or application Water Quality of purification, therefore the formulation containing the above flower domain Bacillus in the above application belong to the protection of the present invention. range.

[0024] 由于本发明首次公开了花域芽孢杆菌作为主要活性成份制成制剂在海参、虾、鲍鱼或鱼等养殖中的应用,因此含上述花域芽孢杆菌的制剂在海参、虾、鲍鱼或鱼等养殖中的应用均属于本发明的保护范围。 [0024] Since the present invention discloses for the first time domain Bacillus flowers use as the main active ingredient formulations prepared in cucumber, shrimp, fish farming abalone, or the like, so that preparations containing the above-described flower domain Bacillus sea cucumber, shrimp, abalone or fish farming applications belong to the scope of the present invention.

[0025] 本发明所述的花域芽孢杆菌在制成任何一种剂型时,均具有在水产养殖中预防疾病、提高饲料转化率或净化养殖水质的作用。 Flower Bacillus domain of [0025] the present invention when formed into any dosage form, each with the prevention of diseases in aquaculture, feed conversion or the water quality purifying breeding. 任何剂型,如果其组份中含有上述花域芽孢杆菌成份制备成制剂,在其包装或说明书等标识上或者在其他任何宣传品上只要注明或提示具有在水产养殖中预防疾病、提高饲料转化率或净化养殖水质的作用,则落入本发明的保护范围之内。 Any dosage form, if its composition containing the above ingredients Bacillus flower field prepared in a preparation, on the packaging or the like as long as the instructions indicate or suggest the identity or having disease prevention in aquaculture on any other promotional materials, feed conversion or purification of water quality in aquaculture, fall within the scope of the present invention.

[0026] 本发明所述的花域芽孢杆菌可以制成兽药、饲料添加剂或养殖水质净化剂。 [0026] Bacillus flower domain of the present invention may be made of veterinary medicine, water purification aquaculture feed additive or agent. 将上述花域芽孢杆菌制成的兽药、饲料添加剂或养殖水质净化剂应用于在水产养殖中预防疾病、提高饲料转化率或净化养殖水质,则落入本发明的保护范围之内。 The above-described flower Bacillus made veterinary field, aquaculture feed additives or water purifying agent used in the prevention of diseases in aquaculture, feed conversion or the water quality purifying breeding, it falls within the scope of the present invention.

具体实施方式 detailed description

[0027]制备例说明:上述已经对花域芽孢杆菌制剂的制备进行说明,这里通过花域芽孢杆菌DHH28保藏编号CGMCCS561,花域芽孢杆菌DHH06保藏编号CGMCC8566颗粒剂为例进行具体说明,其他花域芽孢杆菌菌种制剂的制备方法本领域技术人员通过本实施例很容易掌握,其他剂型的制备方法本领域技术人员通过本实施很容易掌握,在此不再一一叙述说明。 [0027] Preparation Example Description: The above formulation has been prepared flower Bacillus domain will be described here DHH28 Accession Number CGMCCS561 domain by spend, Bacillus subtilis flower domain DHH06 Accession No. CGMCC8566 example specifically described granules, flower fields other Bacillus species formulations prepared by those skilled in the present embodiment is easy to grasp, other dosage forms prepared by those skilled in the present embodiment is easy to grasp, which is not going to describe FIG. 制备方法并不局限于本发明实施例所述,公知的能够达到制备目的的方法均可以,实施例的制备说明只是对本发明的说明,并不是对本发明保护范围的限制。 Preparation methods are not limited to the embodiment of the present invention, the method can achieve the purpose of preparation are well known can be prepared just described embodiment, the present invention is not intended to limit the description of the scope of the present invention.

[0028]制备实施例花域芽孢杆菌活菌粉的制备[0029] 1菌粉的制备和菌种的鉴定 [0028] Preparation Example flowers identified domain viable Bacillus powder [0029] Preparation 1 and the strain sinensis

[0030]采取含有花域芽孢杆菌的样品(海参肠道内容物、海水),然后将样品置于灭菌瓶内,迅速从中取部分样品加入18mL灭菌的稀释液中,充分混匀,在无菌操作台内,进行10-1、 10_2、10_3、10_4,10_5,10—6,10_7 梯度稀释,取l〇-5,l〇-6,10-7 三个稀释梯度,8(TC 水浴13 分钟, 涂布于花域芽孢杆菌选择性单菌落分离固体培养基上,置于厌氧罐内,厌氧30°C下培养48 小时,选择长势良好的单菌落分别接种到花域芽孢杆菌液体扩增培养基中,厌氧30°C下扩增培养4S小时。将所得培养液离心(12000rpm)分离出菌体后,将菌体冷冻真空干燥,调制出干燥菌粉,活菌数均为IX 109CFU/g以上,然后根据《伯杰细菌鉴定手册》、相关细菌鉴定文献和分离菌株进行16S rRNA基因测序后用BLAST与GenBank和RDP数据库中的基因序列进行相似性分析,经鉴定为花域芽孢杆菌DHH28保藏编号CGMCC [0030] Samples were taken (sea cucumber intestinal contents, seawater) containing Bacillus flower field, the samples were then placed sterilized bottle, part of the sample taken from the rapid dilution was added 18mL sterilized, thoroughly mixed, the sterile console, for 10-1, serial dilutions 10_2,10_3,10_4,10_5,10-6,10_7 taken l〇-5, three dilution steps l〇-6,10-7, 8 (TC-water bath 13 minutes, separating the flower field applied selectively Bacillus single colony on solid medium, placed in an anaerobic tank, an anaerobic culture for 48 hours at 30 ° C, good growing selected colonies was inoculated into a single domain flower Bacillus amplification liquid medium, expansion culturing under anaerobic 4S h 30 ° C. after the resultant culture broth was centrifuged (12000 rpm) separating the cells, the cells were freeze-dried in vacuo, to prepare a dried bacterial powder, average viable cell count of IX 109CFU / g or more, and in accordance with "Manual bacterial identification Berger," bacterial relevant similarity BLAST analysis in GenBank and RDP gene databases after separation and identification documents strains 16S rRNA gene sequencing, it was identified as flowers domain Bacillus DHH28 accession number CGMCC 8561,花域芽孢杆菌DHH06保藏编号CGMCC8566。 8561, spent domain Bacillus DHH06 accession number CGMCC8566.

[0031] 2毒性试验 [0031] Toxicity Test 2

[0032] 2_1动物及分组取30只SPF级别小鼠,6〜8周龄,体重15〜19g,随机分入花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组和未给药组,每组10只。 [0032] Animals and 2_1 SPF level packet takes 30 mice, 6 ~ 8 weeks old, weighing 15~19g, randomized domains Bacillus CGMCC8561 group flower, flower fields and non-Bacillus group CGMCC8566 administered groups 10.

[0033] 2.2制备菌液将上述不同的花域芽孢杆菌菌粉分别用纯化水调制为含菌数均为1 X 109CFU/mL 的菌液。 [0033] 2.2 Preparation of the above-described different flowers bacilli Bacillus sinensis domains are modulated with purified water and bacteria are the number 1 X 109CFU / mL bacteria solution.

[0034] 2.3方法各花域芽孢杆菌组和未给药组均给予相同的基础饲料,且饲养条件均一致,各花域芽孢杆菌组每天灌服花域芽孢杆菌菌液0 • 5mL,未给药组每天灌服纯化水0 • 5mL, 饲喂6个月,观察体重及毒性反应。 [0034] 2.3 Method of each flower fields and non-Bacillus group administered group were given the same basic feed and the feeding conditions were the same and each domain flower bacillus took daily gavage domain Bacillus bacteria 0 • 5mL, not given drug group fed purified water per day 0 • 5mL, fed for 6 months, body weight, and toxicity was observed.

[0035] 2.4结果 [0035] 2.4 Results

[0036]各组小鼠均未出现异常情况,未发生振颤、痉挛、运动失调、姿态异常,无眼球突出,排尿正常,皮肤、呼吸正常,无死亡情况,且各花域芽孢杆菌组的体重增加显著高于未给药组(P〈0.05),表明花域芽孢杆菌菌种有促进生长发育作用,说明效果良好,未见中毒反应。 [0036] Groups of mice no abnormal situation, does not occur tremor, spasticity, movement disorders, abnormal posture, no exophthalmos, normal urination, skin, breathing normally, no death, and each group spent domain Bacillus weight gain was significantly higher than non-administered group (P <0.05), show that the domains take Bacillus species have growth promoting effect, indicating good results, and no toxic reactions.

[0037] 3制备成颗粒剂等剂型 [0037] 3 was prepared into a dosage form granules

[0038]根据上述步骤和方法分离鉴定后,经实验检验无毒性,就可将花域芽孢杆菌菌种制成菌粉,然后按照需要添加相关辅料制成各种剂型,优选据花域芽孢杆菌菌粉的活菌数, 按比例添加粘合剂、沸石粉等制成颗粒剂,使活菌数不低于IX 109CFU/g,然后装袋。 [0038] The steps after the separation and identification of the above-described methods, non-toxic checked experimentally, can be made of flower species Bacillus domain sinensis, and related accessories needed to add into various dosage forms, preferably Bacillus domain according to spend the number of viable bacteria powder proportionally adding a binder, the zeolite powder into granules, the number of viable cells of not less than IX 109CFU / g, and bagging.

[0039]应用效果实施例说明:本发明以花域芽孢杆菌DHH28保藏编号CGMCC8561,花域芽孢杆菌DHH06保藏编号CGMCC8566为代表说明花域芽孢杆菌在水产养殖中的应用效果。 Embodiment [0039] Application Example illustrates the effect: In the present invention, the Bacillus flower field DHH28 Accession No. CGMCC8561, flower fields Bacillus DHH06 Accession No. CGMCC8566 flower described as a representative application field effect Bacillus aquaculture.

[0040]应用效果实施例1:花域芽孢杆菌在海参养殖中的应用 Flower in the application domain of Bacillus culture of cucumber: Example 1 [0040] Effect embodiment Applications

[0041] 1材料与方法 [0041] Materials and methods

[0042] 1 • 1材料 [0042] 1 • 1 material

[0043] 1.1.1试验产品 [0043] 1.1.1 Test Product

[0044]花域芽孢杆菌活菌粉剂,活菌数均为1 X l〇9CFU/g以上。 [0044] Bacillus subtilis viable domain flower powder, 1 X number of viable cells are l〇9CFU / g or more.

[0045] 1.1.2试验海参 [0045] 1.1.2 Test cucumber

[0046] 将海参分为3组,每组50头。 [0046] The sea cucumber divided into 3 groups, 50 each.

[0047] 1.2 方法 [0047] 1.2 Methods

[0048] 1.2.1试验时间为30天。 [0048] 1.2.1 Test for 30 days.

[0049] 1 •2.2该试验采取海参常规饲料喂养,试验组每天添加花域芽孢杆菌活菌颗粒剂, 保证水体浓度为1 X l〇7CFU/L以上,对照组不加试验产品。 [0049] 1 • 2.2 The test diet taken cucumber routine, added each day for the test group viable Bacillus domain granules, ensure that the water concentration of 1 X l〇7CFU / L or more, the control group without test product.

[0050] 2试验结果 [0050] 2 Test Results

[0051] 2.1对消化酶的影响 [0051] 2.1 pairs of influence digestive enzymes

[0052] 2.1.1对蛋白酶的影响 Effects [0052] 2.1.1 to proteases

[0053]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将蛋白酶的活性提高31 • 10%、31.31%,差别具有统计学意义(p<〇.〇5),但试验组之间差异无统计学意义(P>〇. 05)。 [0053] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, to improve the activity of the protease 31 • 10%, 31.31%, the difference was statistically significant (p <〇.〇5 ), but not statistically significant (P> square 05) The difference between the test group.

[0054] 2.1.2对淀粉酶的影响 Effects [0054] 2.1.2 amylase

[0055]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将淀粉酶的活性提高25.02%、25.11%,差别具有统计学意义(P<〇.〇5),但试验组之间差异无统计学意义(P>〇.〇5)。 [0055] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, 25.02%, respectively, to increase the activity of amylase, 25.11%, the difference was statistically significant (P <〇.〇5) , but the difference between the experimental group was not statistically significant (P> 〇.〇5).

[0056] 2.2对增强免疫的影响 [0056] 2.2 pairs of enhancing immune

[0057] 2.2.1对超氧化物歧化酶的影响 [0057] 2.2.1 The influence of the superoxide dismutase

[0058]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将超氧化物歧化酶的活性提高11. ie倍、11.61倍,差别具有统计学意义,但试验组之间差异无统计学意义(P>〇. 05)。 [0058] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, superoxide dismutase fold enhanced activity 11. ie, 11.61 times, the difference was statistically significant, but the test no significant difference (P> square 05) between the groups.

[0059] 2.2.2对溶菌酶的影响 Influence [0059] 2.2.2 lysozyme

[0060]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将溶菌酶的活性提高19.03倍、I9.10倍,差别具有统计学意义(p<〇.〇5),但试验组之间差异无统计学意义(P>〇.05)。 [0060] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively lysozyme fold increase in activity 19.03, I9.10 times, the difference was statistically significant (p <〇.〇 5), but no statistical significance (P> 〇.05) difference between test group.

[0061] 2.3对日增重的影响 Effect [0061] The average daily gain of 2.3 pairs

[0062] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将日增重提高31 %、32 %,差别具有统计学意义(P<0 • 05),但试验组之间差异无统计学意义(P>0.05)。 [0062] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, daily gain respectively 31%, 32%, the difference was statistically significant (P <0 • 05), but no significant difference (P> 0.05) between the test group.

[0063] 2.4死亡率 [0063] Mortality 2.4

[0064] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8S66组死亡率均为0,低于对照组的3.15% (P<〇.05),但试验组之间差异无统计学意义(P>〇.05)。 [0064] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8S66 mortality were 0, 3.15% lower than the control group (P <〇.05), but no significant difference between the test group ( P> 〇.05).

[0065] 2.5对养殖水质的影响 Impact of [0065] 2.5 pairs of breeding water quality

[0066] 2.5.1 对pH 的影响 Effects [0066] 2.5.1 on pH

[0067]花域芽孢杆菌0&amp;11〇:8561组、花域芽孢杆菌061^〇:8566组的口唯在6.9〜7.6之间变化,对照组pH值在6 • 2〜7 • 8之间变化,试验组相比对照组变动幅度小,差异有统计学意义(P <0.05),但试验组之间差异无统计学意义(P>〇〇5)。 [0067] Bacillus flower field 0 & amp; 11〇: 8561 group, flower fields 061 ^ Bacillus ○: between 6 • 2~7 • 8 port group 8566 the only change between the control group the pH 6.9~7.6 changes in the test group compared to the control group in small changes, was statistically significant (P <0.05), but no significant difference (P test group> 〇〇5). 2.5.2对亚硝酸盐、氨态氮的影响[0068]各试验组亚硝酸盐、氨态氮低于对照组38% (P<〇.05),但试验组之间差异无统计学意义(P>0.05)。 2.5.2 Effect of nitrite, ammoniacal nitrogen [0068] Each test group of nitrites, ammonia nitrogen than the control group 38% (P <〇.05), but no significant difference between the test group (P> 0.05).

[0069] 3结论 [0069] CONCLUSION

[0070]花域芽孢杆菌能够提高海参日增重、消化酶活性、超氧化物歧化酶活性、溶菌酶活性,并降低死亡率和改善水质,且试验组之间差异没有统计学意义,说明花域芽孢杆菌具有促进海参的生长,提高海参免疫力,防治海参疾病,提高饲料转化率或净化养殖水质的作用[0071]应用效果实施例2:花域芽孢杆菌在虾养殖中的应用[0072] 1材料与方法 [0070] Bacillus flower field can be improved daily gain cucumber, digestive enzyme activity, superoxide dismutase activity, lysozyme activity, and to reduce mortality and improve water quality, and there is no significant difference between the test group, indicating flowers Example Bacillus domain has the effect of promoting the growth of sea cucumber, sea cucumber improve immunity, prevention of cucumber disease and improve feed conversion or purifying breeding water quality [0071] application effect 2: flower application domain Bacillus shrimp in [0072] mATERIALS aND mETHODS

[0073] 1 • 1材料 [0073] 1 • 1 material

[0074] 1.1.1试验产品 [0074] 1.1.1 Test Product

[0075]花域芽孢杆菌活菌粉剂,活菌数均为1 X l〇9CFU/g以上。 [0075] Bacillus subtilis viable domain flower powder, 1 X number of viable cells are l〇9CFU / g or more.

[0076] 1.1.2试验虾 [0076] 1.1.2 Test shrimp

[0077] 将虾分为3组,每组90尾,每组设3个重复。 [0077] The shrimp divided into 3 groups, 90 per group, 3 replications each.

[0078] 1.2 方、法 [0078] 1.2 side, France

[0079] 1.2.1试验时间为30天。 [0079] 1.2.1 Test for 30 days.

[0080] 1.2.2该试验采取虾常规饲料喂养,试验组每天添加花域芽孢杆菌活菌颗粒剂,保证水体浓度为1 X 107CFU/L以上,对照组不加试验产品。 [0080] 1.2.2 General The test diet taken shrimp, added each day for the test group viable Bacillus domain granules, ensure that the water concentration of 1 X 107CFU / L or more, the control group without test product.

[0081] 2试验结果 [0081] 2 Test Results

[0082] 2.1对促进生长的影响 [0082] 2.1 pairs of influence to promote the growth of

[0083]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将体重量提高18.11 %、17.89%,差别具有统计学意义(P<〇. 05),但试验组之间差异无统计学意义(P>0.05)。 [0083] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, 18.11%, respectively, to increase the weight, 17.89%, the difference was statistically significant (P <square. 05), but the test no significant difference (P> 0.05) between groups.

[0084] 2.2对增强免疫的影响 [0084] 2.2 pairs of enhancing immune

[0085] 2.2.1对超氧化物歧化酶的影响 [0085] 2.2.1 The influence of the superoxide dismutase

[0086] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将超氧化物歧化酶的活性提高10.65%、10.78%,差别具有统计学意义,但试验组之间差异无统计学意义(P>〇. 05)。 [0086] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, the activity of superoxide dismutase increase 10.65%, 10.78%, the difference was statistically significant, but the test group no significant difference between (P> square 05).

[0087] 2.2.2对溶菌酶的影响 Influence [0087] 2.2.2 lysozyme

[0088] 结果表明花域芽孢杆菌CGMCC8S61组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将溶菌酶的活性提高9.52%、9 • 48%,差别具有统计学意义(p<〇. 〇5),但试验组之间差异无统计学意义(P>〇.05)。 [0088] The results show that Bacillus CGMCC8S61 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, lysozyme activity increased 9.52%, 9 • 48%, the difference was statistically significant (p <billion. Billion 5), but no statistical significance (P> 〇.05) difference between test group.

[0089] 2.3死亡率 [0089] Mortality 2.3

[0090] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组死亡率分别为2 • 12 %、2 • 31 %,低于对照组的9 • 56% (P<0 • 05),但试验组之间差异无统计学意义(P> 0.05) 〇 [0090] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 mortality is 2 • 12%, 2 • 31%, respectively, 9 • 56% lower than the control group (P <0 • 05), but There was no significant (P> 0.05) between the test groups were square

[0091] 2.4对料重比的影响 [0091] Effect of feed weight ratio of 2.4 pairs

[0092]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组料重比分别为1 • 32、1 • 33,低于对照组的1.98 (P<0.05),但试验组之间差异无统计学意义(P>0.05) 〇[0093] 2.5对养殖水质的影响 [0092] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus feed conversion ratio were set CGMCC8566 1 • 32,1 • 33, lower than 1.98 in the control group (P <0.05), but no difference between experimental group statistically significant (P> 0.05) square Effects [0093] 2.5 pairs of Water quality

[0094] 2.5.1 对pH 的影响 Effects [0094] 2.5.1 on pH

[0095]试验组的pH值在6.6〜7 • 4之间变化,对照组pH值在6.1〜7.9之间变化,试验组相比对照组变动幅度小,差异有统计学意义(P<〇. 05),但试验组之间差异无统计学意义(P> 0 • 05) 〇 [0095] The pH of the test group between 6.6~7 • 4 change, change in pH in the control group, the test group compared to the control group less variation between 6.1~7.9, the difference was statistically significant (P <square. 05), but no statistically significant difference between the experimental group (P> 0 • 05) billion

[0096] 2.5.2对亚硝酸盐、氨态氮的影响 Effects [0096] 2.5.2 nitrite, ammoniacal nitrogen

[0097]各试验组亚硝酸盐、氨态氮低于对照组36% (P<0.05),但试验组之间差异无统计学意义(P>〇.〇5)。 [0097] Each test group nitrite, ammoniacal nitrogen is less than 36% in the control group (P <0.05), but no significant difference (P test group> 〇.〇5).

[0098] 3结论 [0098] CONCLUSION

[00"]花域芽孢杆菌能够提高好体重量、超氧化物歧化酶活性、溶菌酶活性,并降低死亡率、料重比和改善水质,且试验组之间差异没有统计学意义,说明花域芽孢杆菌具有促进虾的生长,提高虾免疫力,防治虾疾病,提高饲料转化率或净化养殖水质的作用。 [00 '] can be improved flower field Bacillus good body weight, superoxide dismutase activity, lysozyme activity, and to reduce mortality, feed conversion ratio and improve water quality, and there is no significant difference between the test group, indicating flowers field has the effect of promoting the growth of Bacillus shrimp, prawns enhance immunity, prevent shrimp disease and improve feed conversion or the water quality purifying breeding.

[0100] 应用效果实施例3:花域芽孢杆菌在鲍鱼养殖中的应用 Flower in the application domain of Bacillus abalone in: Example 3 [0100] Effect embodiment Applications

[0101] 1材料与方法 [0101] Materials and methods

[0102] 1.1材料 [0102] 1.1 Materials

[0103] 1.1.1试验产品 [0103] 1.1.1 Test Product

[0104] 花域芽孢杆菌活菌粉剂,活菌数均为1 X 109CFU/g以上。 [0104] Bacillus subtilis viable powders flower field, number of viable cells were 1 X 109CFU / g or more.

[0105] 1.1.2试验鲍鱼 [0105] 1.1.2 Test abalone

[0106] 将鲍鱼分为3组,每组90头,每组设3个重复。 [0106] abalone divided into 3 groups, 90 per group, 3 replications each.

[0107] 1.2 方、法 [0107] 1.2 side, France

[0108] 1.2.1试验时间为30天。 [0108] 1.2.1 Test for 30 days.

[0109] 1 • 2.2该试验采取鲍鱼常规饲料喂养,试验组每天添加花域芽孢杆菌活菌颗粒剂, 保证水体浓度为1 X 107CFU/L以上,对照组不加试验产品。 [0109] 1 • 2.2 abalone take the test routine diet, added each day for the test group viable Bacillus domain granules, ensure that the water concentration of 1 X 107CFU / L or more, the control group without test product.

[0110] 2试验结果 [0110] 2 Test Results

[0111] 2.1对促进生长的影响 [0111] 2.1 pairs of influence to promote the growth of

[0112]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将体重量提高13.32%、13.50%,差别具有统计学意义史〈〇.〇5),但试验组之间差异无统计学意义(P>〇.〇5)。 [0112] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, 13.32%, respectively, to increase the weight, 13.50%, the difference was statistically significance History <〇.〇5), but the test no significant difference (P> 〇.〇5) between groups.

[0113] 2.2对增强免疫的影响 [0113] 2.2 pairs of enhancing immune

[0114] 2.2.1对超氧化物歧化酶的影响 [0114] 2.2.1 The influence of the superoxide dismutase

[0115]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将超氧化物歧化酶的活性提高15 • 58 %、15 • 69 %,差别具有统计学意义,但试验组之间差异无统计学意义(P>〇. 05)。 [0115] The results show that the flower fields Bacillus CGMCC8561 group, flower fields Bacillus CGMCC8566 group compared with the control group, respectively superoxide dismutase activity increased 15 • 58%, 15 • 69%, the difference was statistically significant, but not statistically significant (P> square 05) The difference between the test group.

[0116] 2.2.2对溶菌酶的影响 Influence [0116] 2.2.2 lysozyme

[0117]结果表明花域芽孢杆菌CGMCCS561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将溶菌酶的活性提高12.23%、12.15%,差别具有统计学意义(P<0.05),但试验组之间差异无统计学意义(P>0.05)。 [0117] The results show that Bacillus CGMCCS561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, lysozyme activity increased 12.23 percent, 12.15 percent, the difference was statistically significant (P <0.05), but the test no significant difference (P> 0.05) between groups.

[0118] 2.3死亡率 [0118] Mortality 2.3

[0119] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组死亡率分别为2.32%、2 • 10 %,低于对照组的9.29% (P<0.05),但试验组之间差异无统计学意义(P> 0.05) 〇 [0119] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 mortality were 2.32%, 2 • 10%, less than 9.29% in the control group (P <0.05), but no difference between experimental group statistically significant (P> 0.05) billion

[0120] 2.4对饲料与体重比的影响 Effects [0120] 2.4 pairs of feed to body weight ratio

[0121] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组料重比分别为1 • 25、1.33,低于对照组的1.88 (P<0.05),但试验组之间差异无统计学意义(P>0.05)。 [0121] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus feed conversion ratio were set CGMCC8566 1 • 25,1.33, 1.88 lower than the control group (P <0.05), but no significant difference between the test group significance (P> 0.05).

[0122] 2.5对养殖水质的影响 Impact of [0122] 2.5 pairs of breeding water quality

[0123] 2.5. IX寸pH的影响 Effects [0123] 2.5. IX inch of pH

[0124] 试验组的pH值在6.5〜7.1之间变化,对照组pH值在6.1〜7.8之间变化,试验组相比对照组变动幅度小,差异有统计学意义(P<0.05),但试验组之间差异无统计学意义(P> 0.05)。 pH value [0124] between the test group 6.5~7.1 change, control change in pH, changes in amplitude compared to the control group Test group between 6.1~7.8 small, it was statistically significant (P <0.05) differences, no significant difference (P> 0.05) between the test group.

[0125] 2.5.2对亚硝酸盐、氨态氮的影响 Effects [0125] 2.5.2 nitrite, ammoniacal nitrogen

[0126] 各试验组亚硝酸盐、氨态氮低于对照组32% (P<0.05),但试验组之间差异无统计学意义(P>〇.〇5)。 [0126] Each test group of nitrites, ammonia nitrogen than the control group 32% (P <0.05), but no significant difference (P test group> 〇.〇5).

[0127] 3 结论 [0127] CONCLUSION

[0128] 花域芽孢杆菌能够提高鲍鱼体重量、超氧化物歧化酶活性、溶菌酶活性,并降低死亡率、料重比和改善水质,且试验组之间差异没有统计学意义,说明花域芽孢杆菌具有促进鲍鱼的生长,提高鲍鱼免疫力,防治鲍鱼疾病,提高饲料转化率或净化养殖水质的作用。 [0128] Bacillus flower field can be improved abalone body weight, superoxide dismutase activity, lysozyme activity, and to reduce mortality, feed conversion ratio and improve water quality, and there is no significant difference between the test group, indicating flower field Bacillus abalone having growth promotion, improve abalone immune system, disease prevention abalone, improved feed conversion or purification of Water quality. [0129] 应用效果实施例4:花域芽孢杆菌在鲈鱼养殖中的应用[0] 1材料与方法 Example 4 [0129] Application of results: Bacillus application domain flower perch aquaculture [0] 1 Materials and methods

[0131] 1_1材料 [0131] Materials 1_1

[0132] 1.1.1试验产品 [0132] 1.1.1 Test Product

[0133]花域芽孢杆菌活菌粉剂,活菌数均为1 X l〇9CFU/g以上。 [0133] Bacillus subtilis viable domain flower powder, 1 X number of viable cells are l〇9CFU / g or more.

[0134] 1.1.2试验鲍鱼 [0134] 1.1.2 Test abalone

[0135] 将鲈鱼分为3组,每组90头,每组设3个重复。 [0135] The bass divided into 3 groups, 90 per group, 3 replications each.

[0136] 1_2方法 [0136] Method 1_2

[0137] 1.2.1试验时间为30天。 [0137] 1.2.1 Test for 30 days.

[0138] 1 • 2.2该试验采取鲈鱼常规饲料喂养,试验组每天添加花域芽孢杆菌活菌颗粒剂, 保证水体浓度为1 X l〇7CFU/L以上,对照组不加试验产品。 [0138] 1 • 2.2 The test diet taken by a conventional bass, added each day for the test group Bacillus subtilis viable domain granules, ensure that the water concentration of 1 X l〇7CFU / L or more, the control group without test product.

[0139] 2试验结果 [0139] 2 Test Results

[0140] 2.1对促进生长的影响 [0140] 2.1 pairs of influence to promote the growth of

[0141]结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将体重量提高3〇_58%、3〇_69%,差别具有统计学意义(p〈〇.〇5),但试验组之间差异无统计学意义(P>〇.〇5)。 [0141] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, to increase the weight% 3〇_58, 3〇_69%, the difference was statistically significant (p <square .〇5), but no statistical significance (P> 〇.〇5) difference between test group.

[0142] 2.2对增强免疫的影响 [0142] 2.2 pairs of enhancing immune

[0143] 2 • 2 • 1对超氧化物歧化酶的影响 [0143] 2 • 2 • 1 influence on superoxide dismutase

[0144] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将超氧化物歧化酶的活性提高13.51 %、13.67 %,差别具有统计学意义,但试验组之间差异无统计学意义(P>〇. 05)。 [0144] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, the activity of superoxide dismutase increase 13.51%, 13.67%, the difference was statistically significant, but the test group no significant difference between (P> square 05).

[0145] 2.2.2对溶菌酶的影响 Influence [0145] 2.2.2 lysozyme

[0146] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组与对照组相比, 分别将溶菌酶的活性提高50.12%、50.32%,差别具有统计学意义(P<0.05),但试验组之间差异无统计学意义(P>〇. 05)。 [0146] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 group compared with the control group, respectively, lysozyme activity increased 50.12 percent, 50.32 percent, the difference was statistically significant (P <0.05), but the test no significant difference (P> square 05) between the groups.

[0147] 2.3死亡率 [0147] Mortality 2.3

[0148] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组死亡率分别为3.12%、3.10 %,低于对照组的10.12 % (P<0.05),但试验组之间差异无统计学意义(P> 0_05) 〇 [0148] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus CGMCC8566 mortality 3.12%, 3.10%, less than 10.12% in the control group (P <0.05), but no significant difference between the test group significance (P> 0_05) billion

[0149] 2.4对饲料与体重比的影响 Effects [0149] 2.4 pairs of feed to body weight ratio

[0150] 结果表明花域芽孢杆菌CGMCC8561组、花域芽孢杆菌CGMCC8566组料重比分别为0.96、0.97,低于对照组的1.56 (P<0.05),但试验组之间差异无统计学意义(P>0.05)。 [0150] The results show that Bacillus CGMCC8561 group flower field, flower fields Bacillus feed weight ratio CGMCC8566 groups were 0.96,0.97, less than 1.56 (P <0.05) in the control group, but no significant difference between the test group ( P> 0.05).

[0151] 2.5对养殖水质的影响 Impact of [0151] 2.5 pairs of breeding water quality

[0152] 2.5.1 对pH 的影响 Effects [0152] 2.5.1 on pH

[0153] 试验组的pH值在6.6〜7.5之间变化,对照组pH值在6.2〜7.8之间变化,试验组相比对照组变动幅度小,差异有统计学意义(P<〇.05),但试验组之间差异无统计学意义(P> 0_05)。 [0153] The pH changes in the test group and control group the pH value changes, the test group compared to the control group less variation between 6.2~7.8 6.6~7.5 between, the difference was statistically significant (P <〇.05) , but not statistically significant (P> 0_05) the difference between the test group.

[0154] 2.5.2对亚硝酸盐、氨态氮的影响 Effects [0154] 2.5.2 nitrite, ammoniacal nitrogen

[0155] 各试验组亚硝酸盐、氨态氮低于对照组35% (P〈0.05),但试验组之间差异无统计学意义(P>〇.〇5)。 [0155] Each test group nitrite, ammoniacal nitrogen is less than 35% in the control group (P <0.05), but no significant difference (P test group> 〇.〇5).

[0156] 3 结论 [0156] CONCLUSION

[0157] 花域芽孢杆菌能够提高鲈鱼体重量、超氧化物歧化酶活性、溶菌酶活性,并降低死亡率、料重比和改善水质,且试验组之间差异没有统计学意义,说明花域芽孢杆菌具有促进鲈鱼的生长,提高鲈鱼免疫力,防治鲈鱼疾病,提高饲料转化率或净化养殖水质的作用。 [0157] Bacillus flower field can be improved bass body weight, superoxide dismutase activity, lysozyme activity, and to reduce mortality, feed conversion ratio and improve water quality, and there is no significant difference between the test group, indicating flower field Bacillus having growth promoting bass, perch improve immunity, disease prevention bass, improved feed conversion or purification of Water quality. [0158] 应用效果实施例5:花域芽孢杆菌在净化海水中的应用 Application field flower Bacillus seawater purification: Example 5 [0158] Effect embodiment Applications

[0159] 取海水,分为三份,其中两份海水分别加入花域芽孢杆菌CGMCC8561、花域芽孢杆菌CGMCC8566,保证每天水体浓度为1 X l〇7CFU/L以上,另份海水为对照组,不加任何活菌, 放置于室温下7天,检测海水,各花域芽孢杆菌组亚硝酸盐、氨态氮低于对照组25% (P< 0.05),但各花域芽孢杆菌组之间差异无统计学意义(P>0.05)。 [0159] Seawater taken, divided into three, two of which were added to water flower domain Bacillus CGMCC8561, flower fields Bacillus CGMCC8566, every day to ensure a concentration of 1 X water l〇7CFU / L or more, the other parts of water as the control group, without any viable allowed to stand for 7 days at room temperature, water is detected, the domain of each flower bacillus nitrite, ammoniacal nitrogen is less than 25% in the control group (P <0.05), but between domains flower bacillus no significant difference (P> 0.05) differences. 提供了良好的养殖环境,预防疾病的发生。 It provides a good breeding environment, prevention of diseases.

[0160] 本发明在实施过程中所使用的微生物菌种已于2013年12月06日在中国微生物菌种保藏管理委员会普通微生物中心(北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所邮编100101)保藏,共2个下述微生物菌种,但本发明所述的花域芽孢杆菌并不局限于这2微生物菌种。 [0160] The present invention Microorganisms in the implementation process was used in Chinese Culture Collection Committee General Microbiology Center (Chaoyang District No. 1 Beichen West Road, December 6, 2013 3 Beijing, Chinese Academy of Sciences Institute of microbial zip code 100101) deposited with a total of two strains of the following microorganisms, but it took Bacillus domain of the present invention is not limited to these two microbial strains.

[0161] (1)分类命名:花域芽抱杆菌Bacillus vallismortis,保存编号8561。 [0161] (1) Category name: Flower field bacillus spores Bacillus vallismortis, save the number 8561.

[0162] ⑵分类命名:花域芽抱杆菌Bacillus vallismortis,保存编号8566。 [0162] ⑵ named Category: Flower field bacillus spores Bacillus vallismortis, save the number 8566.

[0163] 上述2个微生物菌种经该微生物中心检测,检测结果均为存活。 [0163] Microorganisms of the two through the center of the microorganism detection, test results were alive.

Claims (9)

  1. 1. 花域芽孢杆菌在制备水产养殖制剂中的应用,其特征在于所述水产养殖制剂通过增强免疫防治疾病、由保藏编号为CGMCC No.8561的花域芽孢杆菌DHH28和/或保藏编号为CGMCC No.8566的花域芽孢杆菌DHH06制成,其中所述水产养殖为海参、虾、鲍鱼或鱼养殖。 1. Application of the preparation of Bacillus flower domain aquaculture formulation, wherein said formulation aquaculture by enhancing immune disease prevention, the accession number of CGMCC No.8561 flower Bacillus DHH28 domain and / or accession number of CGMCC flower domain of Bacillus No.8566 DHH06 formed, wherein the aquaculture sea cucumber, shrimp, abalone or fish farming.
  2. 2. 按权利要求1所述应用,其特征在于所述水产养殖制剂包括兽药、饲料添加剂。 2. The application according to claim 1, wherein said formulation including veterinary aquaculture, feed additives.
  3. 3. 按权利要求1所述应用,其特征在于花域芽孢杆菌指活的生物个体。 3. The application according to claim 1, characterized in that the flower Bacillus domain refers to a living individual organism.
  4. 4.按权利要求1所述应用,其特征在于所述水产养殖制剂为固体制剂,其包含的总活菌数不低于1 X 106CFU/g。 4. The application according to claim 1, wherein said formulation is a solid formulation of aquaculture, the total number of viable cells comprising not less than 1 X 106CFU / g.
  5. 5. 按权利要求1所述应用,其特征在于所述水产养殖制剂为固体制剂,其包含的总活菌数优选1 X l〇9CFU/g及以上。 5. The application according to claim 1, wherein said formulation is a solid formulation of aquaculture, the total number of viable X-1 preferably comprising l〇9CFU / g and above.
  6. 6. 按权利要求1所述应用,其特征在于所述水产养殖制剂为固体制剂,其包含的总活菌数优选1 X 10nCFU/g及以上。 6. The application according to claim 1, wherein said formulation is a solid formulation of aquaculture, the total number of viable cells which contain preferably 1 X 10nCFU / g and above.
  7. 7.按权利要求1所述应用,其特征在于所述水产养殖制剂为液体制剂,其包含的总活菌数不低于1 X 106CFU/mL。 7. The application according to claim 1, wherein said formulation is a liquid formulation aquaculture, the total number of viable cells comprising not less than 1 X 106CFU / mL.
  8. 8.按权利要求1所述应用,其特征在于所述水产养殖制剂为液体制剂,其包含的总活菌数优选1 X 109CFU/mL及以上。 8. The use according to claim 1, wherein said formulation is a liquid formulation aquaculture, the total number of viable cells which contain preferably 1 X 109CFU / mL and above.
  9. 9.按权利要求1所述应用,其特征在于所述水产养殖制剂为液体制剂,其包含的总活菌数优选1 X 10nCFU/mL及以上。 9. The use according to claim 1, wherein said formulation is a liquid formulation aquaculture, the total number of viable cells which contain preferably 1 X 10nCFU / mL and above.
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