CN115960775A

Movatterモバイル変換

Info

Publication number: CN115960775A
Application number: CN202211531666.0A
Authority: CN
Inventors: 张文龙; 朱坤鹏; 王鸣泽; 李建昊; 张乃生
Original assignee: Jilin University
Current assignee: Jilin University
Priority date: 2023-02-28
Filing date: 2023-02-28
Publication date: 2023-04-14
Anticipated expiration: 2043-02-28
Also published as: CN115960775B

Abstract

The invention provides a lactobacillus salivarius for improving the production performance of goslings and resisting gout, which is preserved in the common microorganism management center of China Committee for culture of microorganisms at 9-16 th 2022, and the preservation address is No. 3 of Xilu No.1 of Beijing City, china, and is named as lactobacillus salivarius (A/A: (A/B))Lactobacillussalivarius) r12, with the preservation number of CGMCC NO.25732; the invention can increase the production performance of goslings and antagonize hyperuricemia caused by high-protein daily rationAnd liver and kidney injury, and has the potential of developing new antibacterial drugs. The feed can increase the weight gain rate of the gosling in 5 days, reduce the feed conversion ratio of the gosling, protect the liver function and kidney function of the gosling, has no harmful influence, can obviously improve the growth performance of the gosling, can be prepared into the feed for feeding the gosling, has the potential of improving the economic benefit of farmers, and has great application prospect.

Description

Translated fromChinese

一株增加雏鹅的生产性能并抗痛风的唾液乳杆菌A strain of Lactobacillus salivarius that increases performance and resists gout in goslings

技术领域technical field

本发明涉及一株新的唾液乳杆菌，进一步提供了一株能够增加雏鹅生产性能并抵抗痛风的唾液乳杆菌，属于微生物技术领域。The invention relates to a new strain of Lactobacillus salivarius, and further provides a strain of Lactobacillus salivarius capable of increasing the production performance of goslings and resisting gout, belonging to the technical field of microorganisms.

背景技术Background technique

唾液乳杆菌属于厚壁菌门，芽孢杆菌纲，乳杆菌目，乳杆菌科，乳杆菌属，是一种过氧化氢酶阴性、革兰氏阳性、非运动性、不产芽孢、专性异型发酵细菌。研究发现，唾液乳杆菌对胃肠道环境具有较好的耐受能力，对人工胃液、肠液和胆盐都有一定的耐受性。这些特性使得唾液乳杆菌具有成为一种在动物饲粮中添加且抗逆性强、能被有效利用的益生菌的潜力。Lactobacillus salivarius belongs to Firmicutes, Bacillus class, Lactobacillus order, Lactobacillus family, Lactobacillus genus, is a catalase-negative, Gram-positive, non-motile, non-spore-forming, obligate isoform fermenting bacteria. Studies have found that Lactobacillus salivarius has a good tolerance to the gastrointestinal environment, and has a certain tolerance to artificial gastric juice, intestinal juice and bile salts. These characteristics make Lactobacillus salivarius have the potential to become a probiotic that can be added to animal diets with strong stress resistance and can be effectively utilized.

唾液乳杆菌广泛存在于人和动物肠道中，具有很强的肠黏附能力，具有抑制胃肠道病原菌繁殖、调节动物免疫机能、降低动物应激、减少动物发病率、调节肠道菌群、预防或减轻腹泻、提高饲料利用效率、促进动物生长、降低料重比等重要作用。基于以上优点，唾液乳杆菌是具有较高开发价值的微生态制剂。Lactobacillus salivarius widely exists in human and animal intestines, has strong intestinal adhesion ability, can inhibit the reproduction of gastrointestinal pathogenic bacteria, regulate animal immune function, reduce animal stress, reduce animal morbidity, regulate intestinal flora, prevent Or reduce diarrhea, improve feed utilization efficiency, promote animal growth, reduce feed-to-weight ratio and other important functions. Based on the above advantages, Lactobacillus salivarius is a microecological preparation with high development value.

发明内容Contents of the invention

本发明公开了一株能够增加雏鹅生产性能并抵抗痛风的唾液乳杆菌，是从健康雏鹅盲肠粪便中分离获得的一个新菌种，能够增加雏鹅生产性能并降低尿酸，具有开发新发酵饲料的潜力。The invention discloses a strain of Lactobacillus salivarius that can increase the production performance of goslings and resist gout. It is a new bacterial strain isolated from the cecum feces of healthy goslings. It can increase the production performance of goslings and reduce uric acid. It has the ability to develop new fermentation feed potential.

本发明提供了一株能够增加雏鹅生产性能并抵抗痛风的唾液乳杆菌，保藏于中国微生物菌种保藏委员会普通微生物管理中心，保藏地址为中国北京市北辰西路1号院3号，保藏日期2022年9月16日，保藏编号为CGMCC NO.25732，命名为：唾液乳杆菌（Lactobacillus salivarius）r12。The invention provides a strain of Lactobacillus salivarius capable of increasing the production performance of goslings and resisting gout, which is preserved in the General Microorganism Management Center of China Committee for the Preservation of Microorganisms, the preservation address is No. 3, No. 1 Yard, Beichen West Road, Beijing, China, and the preservation date is On September 16, 2022, the deposit number was CGMCC NO.25732, named:Lactobacillus salivarius r12.

本发明所述的唾液乳杆菌（Lactobacillus salivarius）r12在制备雏鹅饲料添加剂中的用途，活菌数不低于1×10⁹CFU/mL或1×10⁹CFU/g。The use ofLactobacillus salivarius r12 of the present invention in the preparation of gosling feed additives, the number of live bacteria is not less than 1×10⁹ CFU/mL or 1×10⁹ CFU/g.

本发明所述的唾液乳杆菌（Lactobacillus salivarius）r12在拮抗雏鹅用高蛋白饲料引起的痛风中的应用。The application ofthe Lactobacillus salivarius r12 described in the present invention in antagonizing the gout caused by high-protein feed for goslings.

本发明所述的唾液乳杆菌（Lactobacillus salivarius）r12在制备降低雏鹅血尿酸制剂中的应用。The application ofthe Lactobacillus salivarius r12 described in the present invention in the preparation of a preparation for reducing blood uric acid in goslings.

本发明所述的一株能够增加雏鹅生产性能并抵抗雏鹅痛风的唾液乳杆菌的筛选及培养方法，具体步骤为：A method for screening and cultivating Lactobacillus salivarius that can increase the production performance of goslings and resist goose goose in the present invention, the specific steps are:

乳杆菌选择性培养基（LBS琼脂）：蛋白胨10.0g/L，牛肉浸粉10.0g/L，酵母浸粉2.0g/L，磷酸氢二钾2.0g/L，乙酸钠5.0g/L，枸橼酸三铵2.0g/L，硫酸镁0.2g/L，硫酸锰0.05g/L，吐温801.0g/L，葡萄糖20.0g/L，碳酸钙5.0g/L，琼脂25.0g/L，pH值6.0 - 6.5；采集健康雏鹅盲肠粪便样本进行菌株的分离培养：纯化得到本发明唾液乳杆菌Lactobacillus reuteri，革兰氏染色阳性；将唾液乳杆菌划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下再静置6h，得到菌液；在菌液中加入总体积20-30%的甘油放置于-80℃保存。Lactobacillus selective medium (LBS agar): peptone 10.0g/L, beef extract powder 10.0g/L, yeast extract powder 2.0g/L, dipotassium hydrogen phosphate 2.0g/L, sodium acetate 5.0g/L, citrate Triammonium citrate 2.0g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.05g/L, Tween 801.0g/L, glucose 20.0g/L, calcium carbonate 5.0g/L, agar 25.0g/L, pH The value is 6.0 - 6.5; collect healthy gosling cecal feces samples for isolation and culture of strains:Lactobacillus salivarius reuteri of the present invention is purified, and Gram staining is positive; Lactobacillus salivarius is streaked on the MRS solid medium at 37°C Cultivate at low temperature for 48 hours to obtain a single colony; pick a single colony and inoculate it in the MRS liquid medium, and let it stand for 6 hours at 37°C to obtain a bacterial liquid; add 20-30% of the total volume of glycerol to the bacterial liquid and place it at -80 Store at ℃.

将本发明所述筛选的菌株测序得到的序列在NCBI中进行核酸序列比对，结果显示菌株为唾液乳杆菌，命名为唾液乳杆菌Lactobacillus salivarius r12。The sequences obtained by sequencing the screened strains of the present invention were compared in NCBI, and the result showed that the strain was Lactobacillus salivarius, namedLactobacillus salivarius r12.

所述唾液乳杆菌Lactobacillus salivarius r12的菌体特征：在培养基上形成形状钝圆、边缘整的乳白色菌落。The thalline characteristics ofthe Lactobacillus salivarius r12: milky white colonies with obtuse round shape and neat edges are formed on the culture medium.

所述唾液乳杆菌Lactobacillus salivarius r12的生长特征：该菌株为革兰式阳性兼性厌氧菌，在温度37℃生长最佳。The growth characteristics ofLactobacillus salivarius r12: the bacterial strain is a Gram-positive facultative anaerobic bacterium, and grows best at a temperature of 37°C.

本发明所述的唾液乳杆菌在制备增加雏鹅体重的饲料中的用途，饲料中唾液乳杆菌活菌数不低于1×10⁹CFU/mL或1×10⁹CFU/g。The use of the Lactobacillus salivarius described in the present invention in the preparation of feed for increasing the weight of goslings, the number of viable Lactobacillus salivarius in the feed is not less than 1×10⁹ CFU/mL or 1×10⁹ CFU/g.

本发明的积极效果在于：The positive effects of the present invention are:

筛选出了一株新的菌株品种----唾液乳杆菌r12，能够降低雏鹅血尿酸，能够增加雏鹅的平均体重，而对雏鹅的肝功、肾功无有害影响，能够显著提高雏鹅的生长性能，可以配制成饲料用于饲养雏鹅，有着提升养殖户经济效益的潜力，具有巨大的应用前景。A new strain of strain Lactobacillus salivarius r12 was screened out, which can reduce blood uric acid of goslings and increase the average body weight of goslings, but has no harmful effect on the liver function and kidney function of goslings, and can significantly improve The growth performance of goslings can be formulated into feed for raising goslings, which has the potential to improve the economic benefits of farmers and has great application prospects.

附图说明Description of drawings

图1为本发明两组雏鹅的平均5日增重对比图和料肉比；Fig. 1 is the average 5 day weight gain contrast figure and feed-to-meat ratio of two groups of goslings of the present invention;

图2本发明为两组雏鹅的血尿酸；肝功：血清谷丙转氨酶、血清谷草转氨酶；肾功：血清尿素氮、血清肌酐；肝脏、肾脏组织切片的对比图；Fig. 2 present invention is the blood uric acid of two groups of goslings; Liver function: serum alanine aminotransferase, serum aspartate aminotransferase; Kidney function: serum urea nitrogen, serum creatinine; Comparison diagram of liver and kidney tissue sections;

图3为本发明两组雏鹅的肝脏黄嘌呤氧化酶，腺苷酸脱氨酶的转录水平；肾脏ATP结合超家族G成员2和葡萄糖转运体9的转录水平对比图。Fig. 3 is a comparison chart of transcription levels of liver xanthine oxidase and adenylate deaminase of two groups of goslings of the present invention; transcription levels of kidney ATP-binding superfamily G member 2 and glucose transporter 9.

具体实施方式Detailed ways

下面由特定的具体实施例进一步说明本发明，所描述的实施例是本发明的一部分实施例，而不是全部实施例，但也并不因此将本发明限制在所述的实施例范围之中。下述实施例中未注明具体条件的实验方法，按照常规方法进行，或按照商品说明书进行。The present invention is further illustrated by specific specific embodiments below, and the described embodiments are part of the embodiments of the present invention, rather than all embodiments, but the present invention is not limited to the scope of the described embodiments. For the experimental methods that do not specify specific conditions in the following examples, carry out according to conventional methods, or carry out according to the product instructions.

实施例1Example 1

MRS肉汤培养基：蛋白胨10.0g/L，牛肉粉8.0g/L，酵母粉4.0g/L，葡萄糖20.0g/L，磷酸氢二钾2.0g/L，柠檬酸氢二铵2.0g/L，乙酸钠5.0g/L，硫酸镁0.2g/L，硫酸锰0.04g/L，吐温801.0g/L，pH值5.7±0.2；MRS broth medium: peptone 10.0g/L, beef powder 8.0g/L, yeast powder 4.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 2.0g/L, diammonium hydrogen citrate 2.0g/L , sodium acetate 5.0g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.04g/L, Tween 801.0g/L, pH value 5.7±0.2;

MRS固体培养基：蛋白胨10.0g/L，牛肉粉8.0g/L，酵母粉4.0g/L，葡萄糖20.0g/L，磷酸氢二钾2.0g/L，柠檬酸氢二铵2.0g/L，乙酸钠5.0g/L，硫酸镁0.2g/L，硫酸锰0.04g/L，琼脂14.0g/L，吐温801.0g/L，pH值6.5±0.2；MRS solid medium: peptone 10.0g/L, beef powder 8.0g/L, yeast powder 4.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 2.0g/L, diammonium hydrogen citrate 2.0g/L, Sodium acetate 5.0g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.04g/L, agar 14.0g/L, Tween 801.0g/L, pH 6.5±0.2;

本发明唾液乳杆菌r12菌悬液的制备方法如下：The preparation method of Lactobacillus salivarius r12 bacterial suspension of the present invention is as follows:

将唾液乳杆菌划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下再静置6h，得到菌液；在菌液中加入总体积20-30%的甘油放置于-80℃保存；Streak Lactobacillus salivarius on the MRS solid medium, culture at 37°C for 48 hours to obtain a single colony; pick a single colony and inoculate it in the MRS liquid medium, and then stand it at 37°C for 6 hours to obtain a bacterial solution; Add 20-30% of the total volume of glycerol to the bacterial solution and store at -80°C;

唾液乳杆菌r12的分离Isolation of Lactobacillus salivarius r12

1、样品采集1. Sample collection

采集辽阳市某养鹅场健康雏鹅盲肠粪便样本，样本置于装有总体积20-30%甘油的离心管中，保存于装有冰袋的保温盒中，带回实验室后迅速置于-80℃冰箱待分离筛选。The cecal feces samples of healthy goslings were collected from a goose farm in Liaoyang City. The samples were placed in a centrifuge tube with a total volume of 20-30% glycerol, stored in an insulated box with ice packs, and brought back to the laboratory. Immediately placed in- 80 ℃ refrigerator to be separated and screened.

2、菌株的分离培养2. Isolation and cultivation of bacterial strains

（1）稀释涂布：取0.5g左右的保存于20-30%甘油的内容物在无菌环境下加入装有4.5mL生理盐水的10mL离心管中，得到10^-1稀释液，重复上述稀释步骤，依次得到10^-2、10^-3、10^-4、10^-5、10^-6、10^-7、10^-8稀释液；(1) Dilution and coating: Take about 0.5g of the content stored in 20-30% glycerol and add it into a 10mL centrifuge tube containing 4.5mL of normal saline under a sterile environment to obtain a 10^-1 dilution, repeat the above dilution Step, sequentially obtain 10^-2 , 10^-3 , 10^-4 , 10^-5 , 10^-6 , 10^-7 , 10^-8 dilutions;

（2）划线培养：用接种环沾取步骤(1)中10^-1、10^-2、10^-3、10^-4四个梯度稀释液于乳杆菌选择性培养基上划线，至37℃条件下培养48h，得到划线平板；(2) Streak culture: Use an inoculation loop to dip four gradient dilutions of 10^-1 , 10^-2 , 10^-3 , and 10^-4 in step (1) and streak on the Lactobacillus selective medium until 37 Cultivate at ℃ for 48 hours to obtain a streaked plate;

（3）涂布培养：分别吸取50μL步骤（1）中10^-5、10^-6、10^-7、10^-8四个梯度稀释液于乳杆菌选择性培养基上，用涂布棒涂布均匀，至37℃条件下培养48h，得到稀释涂布平板；(3) Coating culture: pipette 50 μL of four gradient dilutions of 10^-5 , 10^-6 , 10^-7 , and 10^-8 in step (1) respectively on the Lactobacillus selective medium, and spread with a coating rod Evenly, culture at 37°C for 48 hours to obtain a diluted coating plate;

（4）菌液培养：挑取步骤（2）中划线平板、（3）中稀释涂布平板的单菌落分别接种于MRS液体培养基中，至37℃温箱中静止6h，待菌落数为1×10⁹CFU/mL后加入总体积20%-30%的甘油放置于-80℃保存；(4) Bacterial liquid culture: Pick single colonies from the streaked plate in step (2) and the diluted coating plate in (3) and inoculate them in MRS liquid medium respectively, and let them rest in a 37°C incubator for 6 hours. After reaching 1×10⁹ CFU/mL, add glycerol with a total volume of 20%-30% and store at -80°C;

3、菌种的鉴定3. Identification of strains

将步骤2（4）的菌液送吉林省库美生物科技有限公司基因测序；将测序的拼接序列上传到NCBI的BLAST(http://www.ncbi.nlm.nih.gov/BLAST)进行种属确认；比对结果发现为唾液乳杆菌Lactobacillus salivarius，其16S rDNA扩增序列SEQ ID NO.1所示：Send the bacterial solution in step 2 (4) to Jilin Province Kumei Biotechnology Co., Ltd. for gene sequencing; upload the sequenced spliced sequence to BLAST (http://www.ncbi.nlm.nih.gov/BLAST) of NCBI for seeding The genus was confirmed; the result of comparison was found to beLactobacillus salivarius , whose 16S rDNA amplification sequence is shown in SEQ ID NO.1:

4、菌种的保藏4. Preservation of strains

本发明所述的菌株已于2022年9月16日，保藏于中国微生物菌种保藏委员会普通微生物管理中心，保藏地址为中国北京市北辰西路1号院3号，命名为：唾液乳杆菌（Lactobacillus salivarius） r12，保藏编号为CGMCC NO.25732。The bacterial strain described in the present invention has been preserved in the General Microorganism Management Center of the China Microbial Strains Preservation Committee on September 16, 2022. The preservation address is No. 3, No. 1, Beichen West Road, Beijing, China, and it is named: Lactobacillus salivarius (Lactobacillus salivarius) r12, the deposit number is CGMCC NO.25732.

本发明所述的唾液乳杆菌r12具有下列性质：Lactobacillus salivarius r12 of the present invention has the following properties:

1）能够增加雏鹅的体重增长率和料肉比；1) It can increase the weight growth rate and feed-to-meat ratio of goslings;

2）拮抗高蛋白饲料对雏鹅的肝功、肾功的有害影响；2) Antagonize the harmful effects of high-protein feed on the liver function and kidney function of goslings;

3）提高雏鹅的生长性能，有着提升养殖户经济效益的潜力。3) Improving the growth performance of goslings has the potential to improve the economic benefits of farmers.

由上述实施例可以看出，本发明提供了一株乳杆菌新菌种，并对新菌种的基本特性、培养方法和应用进行了说明，但也可以看出，在本发明基础上，可以对之做出一些修改，因此，在不偏离本发明精神和原则的基础上所做的修改或改进，均属于本发明要求保护的范围。As can be seen from the foregoing examples, the present invention provides a new strain of Lactobacillus, and the basic characteristics, culture methods and applications of the new strain have been described, but it can also be seen that on the basis of the present invention, Some modifications are made to it, therefore, the modifications or improvements made on the basis of not departing from the spirit and principle of the present invention all belong to the protection scope of the present invention.

实验例1Experimental example 1

唾液乳杆菌r12对雏鹅生长性能的影响Effects of Lactobacillus salivarius r12 on growth performance of goslings

1、唾液乳杆菌r12菌液的制备1. Preparation of Lactobacillus salivarius r12 bacterial liquid

将实施例1获得的唾液乳杆菌r12划线于MRS固体培养基中，37℃条件下培养48h，得到单菌落；挑取MRS固体培养基上的单菌落接种于MRS液体培养基中，至37℃温箱中静置6h，得到菌液；在菌液中加入总体积20-30%的甘油放置于-80℃保存。实验之前取唾液乳杆菌r12悬浮液离心，倒掉上清液后，用两倍体积的无菌生理盐水洗涤重悬，得到唾液乳杆菌r12菌液(1×109CFU/mL)；Streak the Lactobacillus salivarius r12 obtained in Example 1 on the MRS solid medium, culture it at 37°C for 48 hours, and obtain a single colony; pick the single colony on the MRS solid medium and inoculate it in the MRS liquid medium, until 37°C ℃ incubator for 6 hours to obtain the bacterial liquid; add 20-30% of the total volume of glycerol to the bacterial liquid and store at -80°C. Before the experiment, the Lactobacillus salivarius r12 suspension was centrifuged, and after the supernatant was discarded, it was washed and resuspended with twice the volume of sterile saline to obtain the Lactobacillus salivarius r12 bacterial liquid (1×109 CFU/mL);

2、实验动物2. Experimental animals

1日龄雏鹅，饲养初期用暖灯使温度保持37℃，一周后保持室温即可，动物房内一直保持干净、通风；For 1-day-old goslings, use warm lamps to keep the temperature at 37°C at the beginning of breeding, and keep the room temperature after one week. Keep the animal room clean and ventilated;

3、实验分组3. Experimental grouping

将1日龄健康雏鹅40只，随机分为4组，每组10只，4组分别为：正常蛋白组，正常蛋白唾液乳杆菌r12干预组，高蛋白组和高蛋白唾液乳杆菌r12干预组；Forty 1-day-old healthy goslings were randomly divided into 4 groups, 10 in each group. The 4 groups were: normal protein group, normal protein Lactobacillus salivarius r12 intervention group, high protein group and high protein Lactobacillus salivarius r12 intervention Group;

4、实验内容4. Experimental content

（1）正常蛋白组：饲喂正常蛋白日粮；(1) Normal protein group: fed normal protein diet;

（2）正常蛋白唾液乳杆菌r12干预组：饲喂正常蛋白日粮，将唾液乳杆菌r12离心后倒掉上清，加入两倍体积的无菌生理盐水混匀饲喂给雏鹅；(2) Normal protein Lactobacillus salivarius r12 intervention group: fed a normal protein diet, centrifuged Lactobacillus salivarius r12, poured off the supernatant, added twice the volume of sterile saline, mixed and fed to the goslings;

（3）高蛋白组：饲喂高蛋白日粮；(3) High protein group: fed high protein diet;

（4）高蛋白唾液乳杆菌r12干预组：饲喂高蛋白日粮，将唾液乳杆菌r12离心后倒掉上清，加入两倍体积的无菌生理盐水混匀饲喂给雏鹅；(4) High-protein Lactobacillus salivarius r12 intervention group: fed a high-protein diet, centrifuged Lactobacillus salivarius r12, poured off the supernatant, added twice the volume of sterile saline, mixed and fed to the goslings;

（5）每天上午对4组雏鹅的体重和耗粮量进行称量并记录；(5) Weigh and record the body weight and grain consumption of the 4 groups of goslings every morning;

（6）实验周期为25天；(6) The experimental period is 25 days;

5、实验结果5. Experimental results

如图1所示，与正常蛋白组相比，唾液乳杆菌r12能够增加雏鹅的增重速度和料肉比。As shown in Figure 1, compared with the normal protein group, Lactobacillus salivarius r12 can increase the weight gain rate and feed-to-meat ratio of goslings.

实验例2Experimental example 2

唾液乳杆菌r12对雏鹅血尿酸和肝脏、肾脏的影响，具体步骤如下：The effect of Lactobacillus salivarius r12 on gosling blood uric acid, liver and kidney, the specific steps are as follows:

1、唾液乳杆菌r12菌液的制备同实验例1；1. The preparation of the Lactobacillus salivarius r12 bacterial liquid is the same as that of Experimental Example 1;

2、实验动物同实验例1；2. Experimental animals are the same as in Experimental Example 1;

3、实验分组同实验例1；3. The experimental grouping is the same as the experimental example 1;

4、实验内容4. Experimental content

（5）实验周期为25天；(5) The experimental period is 25 days;

（6）在第26天处死雏鹅并收集4组雏鹅的血、肝脏以及肾脏组织，将一部分肝脏、肾脏组织做成切片，并检测肝脏相关指标：血清尿酸（Serum UA）、血清谷丙转氨酶（SerumALT）、血清谷草转氨酶（Serum AST）；肾脏相关指标：血清尿素氮（Serum BUN）、血清肌酐（Serum CREA）；(6) The goslings were killed on the 26th day and the blood, liver and kidney tissues of four groups of goslings were collected, a part of the liver and kidney tissues were sliced, and liver-related indicators were detected: serum uric acid (Serum UA), serum glutamic acid Transaminase (SerumALT), serum aspartate aminotransferase (Serum AST); kidney-related indicators: serum urea nitrogen (Serum BUN), serum creatinine (Serum CREA);

5、实验结果5. Experimental results

如图2所示，高蛋白组与正常蛋白组和高蛋白组与高蛋白唾液乳杆菌r12干预组雏鹅的血清尿酸、血清谷丙转氨酶、血清谷草转氨酶、血清尿素氮、血清肌酐的结果差异显著；高蛋白组与正常蛋白组和高蛋白组与高蛋白唾液乳杆菌r12干预组雏鹅的肝脏、肾脏组织的切片没有差异显著，说明高蛋白饲料对肝肾功能有损伤而唾液乳杆菌r12可以拮抗这种损伤且对雏鹅的肝功、肾功无有害影响。As shown in Figure 2, the results of serum uric acid, serum alanine aminotransferase, serum aspartate aminotransferase, serum urea nitrogen, and serum creatinine were different between the high protein group and the normal protein group, and between the high protein group and the high protein Lactobacillus salivarius r12 intervention group Significantly; there was no significant difference between high protein group and normal protein group and high protein group and high protein L. It can antagonize this damage and has no harmful effect on the liver function and kidney function of goslings.

实验例3Experimental example 3

唾液乳杆菌r12对雏鹅肝脏、肾脏代谢能力影响，具体步骤如下：The effect of Lactobacillus salivarius r12 on the liver and kidney metabolism of goslings, the specific steps are as follows:

4、实验内容4. Experimental content

（5）实验周期为25天；(5) The experimental period is 25 days;

（6）在第26天处死雏鹅并收集4组雏鹅肝脏以及肾脏组织，将肝脏、肾脏组织在液氮中研磨提取RNA，反转录成cDNA，进行荧光定量pcr检测相关指标：腺苷脱氨酶（ADA）、黄嘌呤氧化酶（XOD）、ATP结合核超家族G成员2（ABCG2）、葡萄糖转运体9（GLUT9）；(6) Kill the goslings on the 26th day and collect the liver and kidney tissues of four groups of goslings, grind the liver and kidney tissues in liquid nitrogen to extract RNA, reverse transcribe into cDNA, and perform fluorescent quantitative PCR detection Related indicators: adenosine Deaminase (ADA), xanthine oxidase (XOD), ATP-binding nuclear superfamily G member 2 (ABCG2), glucose transporter 9 (GLUT9);

5、实验结果5. Experimental results

在肝脏中，各组关于嘌呤产生相关的酶转录水平变换较为混乱，但在肾脏中，高蛋白组与尿酸转运相关的蛋白转录显著降低，而高蛋白唾液乳杆菌r12干预组拮抗了这种变化，提升了相关转运蛋白的转录。In the liver, the changes in the transcript levels of enzymes related to purine production in each group were chaotic, but in the kidney, the transcription of proteins related to uric acid transport was significantly reduced in the high-protein group, and the high-protein Lactobacillus salivarius r12 intervention group antagonized this change , which increases the transcription of the associated transporter.

Claims

Translated fromChinese

1.一种唾液乳杆菌（Lactobacillus salivarius）r12，保藏于中国微生物菌种保藏委员会普通微生物管理中心，保藏地址为中国北京市北辰西路1号院3号，保藏日期2022年9月16日，保藏编号为CGMCC NO. 25732。1. ALactobacillus salivarius r12, preserved in the General Microorganism Management Center of the China Committee for the Collection of Microorganisms, the preservation address is No. 3, No. 1, Beichen West Road, Beijing, China, and the preservation date is September 16, 2022. The deposit number is CGMCC NO. 25732.

2.如权利要求1所述的唾液乳杆菌（Lactobacillus salivarius）r12在制备抗痛风雏鹅饲料添加剂中的用途，活菌数不低于1×10⁹CFU/mL或1×10⁹CFU/g。2. The use ofLactobacillus salivarius r12 according to claim 1 in the preparation of anti-gout goose feed additives, the number of live bacteria is not less than 1×10⁹ CFU/mL or 1×10⁹ CFU/g .

3.如权利要求1所述的唾液乳杆菌（Lactobacillus salivarius）r12在制备雏鹅用拮抗高蛋白饲料中的应用。3. The application ofLactobacillus salivarius r12 as claimed in claim 1 in the preparation of antagonistic high-protein feed for goslings.

4.如权利要求1所述的唾液乳杆菌（Lactobacillus salivarius）r12在制备降低雏鹅血尿酸制剂中的应用。4. The application ofLactobacillus salivarius r12 as claimed in claim 1 in the preparation of preparations for reducing blood uric acid in goslings.