不同氮源對(duì)豬腸道微生物多樣性及主要產(chǎn)蛋白酶菌株的影響
發(fā)布時(shí)間：2018-10-31 19:21
【摘要】：為了探究低蛋白日糧中,不同蛋白源對(duì)豬腸道微生物的影響,本研究以棉籽粕、魚粉和玉米胚芽粕等作為主要蛋白來(lái)源,考察了低蛋白日糧對(duì)仔豬和育肥豬腸道微生物多樣性和主要產(chǎn)蛋白酶菌株的影響。本研究共包括三個(gè)試驗(yàn)。試驗(yàn)一考察低蛋白日糧不同氮源對(duì)斷奶仔豬小腸微生物的影響。將48頭斷奶仔豬(14.62±1.46 kg),隨機(jī)分為4組,每組3個(gè)重復(fù),每個(gè)重復(fù)4頭仔豬,仔豬采用單籠飼養(yǎng)。試驗(yàn)處理為對(duì)照組(SBM組)、棉籽粕(CSM)、豆粕-棉籽粕(SC)和魚粉(FM)組,其中CSM組和SC組日糧中棉籽粕分別替代對(duì)照組豆粕提供粗蛋白的100%和50%。日糧以凈能體系配制,補(bǔ)充平衡各組必需氨基酸。飼養(yǎng)試驗(yàn)結(jié)束時(shí),每個(gè)重復(fù)隨機(jī)選取1頭仔豬進(jìn)行屠宰、取樣利用Illumina Miseq測(cè)序技術(shù)測(cè)定十二指腸、空腸前段、中段、后段和回腸微生物多樣性。研究結(jié)果表明,不同蛋白源組仔豬小腸微生物多樣性和主要微生物組成差異均較大。其中,Firmicutes、Proteobacteria、Bacteroidetes和Acidobacteria在各組豐度均較高,添加棉籽粕兩組中有益菌Lactobacillus和Clostridium豐度較其他組更高,而FM組中Escherichia/Shigella豐度較高。由此得出,不同蛋白源對(duì)仔豬小腸微生物多樣性有較大影響,棉籽粕替代豆粕日糧提高了仔豬小腸有益菌比例。試驗(yàn)二研究了低蛋白日糧不同氮源對(duì)育肥豬生長(zhǎng)性能和腸道微生物的影響。選取36頭杜長(zhǎng)大育肥閹公豬(58.65±3.71 kg)隨機(jī)分在4組中,每個(gè)處理3個(gè)重復(fù),每個(gè)重復(fù)3頭豬。處理組為對(duì)照組(SBM15)組和三個(gè)低蛋白(11%CP)日糧組,其中豆粕(SBM11)、棉籽粕和玉米胚芽粕50%等氮替代豆粕(SCCM)、棉籽粕和玉米胚芽粕100%等氮替代豆粕(CCM)分別作為三種低蛋白日糧中主要蛋白源。日糧以凈能體系配制,補(bǔ)充平衡各組必需氨基酸。經(jīng)過(guò)28 d飼喂后,取回腸內(nèi)容物和糞便作為腸道微生物分析樣本,通過(guò)Illumina Miseq測(cè)序技術(shù)測(cè)定樣本微生物多樣性。結(jié)果顯示,SBM11與CCM組生長(zhǎng)性能顯著低于SBM15組(P0.05),但SCCM組與SBM15組生長(zhǎng)性能沒(méi)有顯著差異(P0.05)。對(duì)16SrRNA V3-V4區(qū)進(jìn)行擴(kuò)增測(cè)序后發(fā)現(xiàn),SCCM組日糧顯著增加了育肥豬腸道微生物多樣性,并提高了有益菌(如Lactobacillus和Bifidobacterium)豐度。除此之外,Ruminococcus、Prevotellaceae和Bacteroides相對(duì)豐度在低蛋白日糧組中也有顯著提升。因此,豆粕、棉籽粕和玉米胚芽粕作為育肥豬日糧主要蛋白源即使在低蛋白的情況下,可以調(diào)節(jié)腸道微生物,維持育肥豬的正常生長(zhǎng)。試驗(yàn)三從健康仔豬小腸內(nèi)篩選一株產(chǎn)蛋白酶菌2L6,經(jīng)形態(tài)、生理生化和分子生物學(xué)鑒定確定該菌株為蠟樣芽胞桿菌(Bacillus cereus)。分別在豆粕、棉籽粕、豆粕-棉籽粕和魚粉發(fā)酵培養(yǎng)基中對(duì)其進(jìn)行發(fā)酵培養(yǎng),并在不同時(shí)間檢測(cè)蛋白酶活性和底物蛋白消化率。結(jié)果表明,菌株2L6在不同發(fā)酵培養(yǎng)基中蛋白酶活性及底物蛋白消化率有較大差異。其中,豆粕-棉籽粕混合發(fā)酵培養(yǎng)基中蛋白酶活性在相對(duì)較短時(shí)間內(nèi)即達(dá)到最大值21.31±0.09U/m L,而底物蛋白消化率也在36 h即達(dá)到最大值64.42%(P0.01)。因此,豆粕-棉籽粕混合蛋白源可較大提高仔豬小腸主要蛋白酶菌(Bacillus cereus strain 2L6)產(chǎn)蛋白酶活力及其對(duì)蛋白質(zhì)利用能力。綜上所述,低蛋白日糧中以棉籽粕、玉米胚芽粕等雜粕作為主要蛋白來(lái)源,可以改善仔豬和育肥豬的腸道微生物組成,增高有益菌比例,提高腸道中主要產(chǎn)蛋白酶菌株活性,可作為低蛋白日糧中豆粕的合理替代選擇。
[Abstract]:In order to explore the effect of different protein sources on the intestinal microorganisms in pigs, the effects of the diets on the microbial diversity and the major protease-producing strains of piglets and fattening pigs were studied by using cottonseed meal, fish meal and corn germ meal as the main source of protein. This study consisted of three trials. Effects of different nitrogen sources on small intestinal microorganisms in weanling piglets. Forty-eight weanling piglets (14.62. 1.46 kg) were randomly divided into 4 groups, 3 replicates in each group, 4 piglets were repeated, and the piglets were fed with single cage. The test was treated as control group (SBM group), cottonseed meal (CSM), soybean meal-cottonseed meal (SC) and fish meal (FM) group, among which, the cottonseed meal in the CSM group and SC group was 100% and 50% of the crude protein in the control group respectively. Daily grain is formulated in net energy system to supplement essential amino acids in each group. At the end of the breeding trial, 1 piglets were randomly selected for slaughter and sampling was performed using the Illumina Miseq sequencing technique to determine the diversity of the duodenum, the anterior segment, the middle segment, the posterior segment, and the ileum. The results showed that the diversity of small intestine and the main microbial composition of piglets in different protein source groups were different. Among them, Firmicutes, Proteobacteria, Bacteroidites and Acidobacter were higher in each group, and the abundance of Lactobacillus and Clostridia was higher in the two groups of cottonseed meal, while the abundance of Escherrez/ Shigella in FM group was higher. The results showed that different protein sources had a great influence on the microbial diversity of small intestine of piglets. The effects of different nitrogen sources on the growth and intestinal microorganisms of fattening pigs were studied. Thirty-six (58. 65/ 3. 71 kg) pigs were randomly divided into 4 groups, 3 replicates per process and 3 pigs per replicate. The treatment group was a control group (SBM15) group and three diets (11% CP) diet group, in which 50% of soybean meal (SBM15), cottonseed meal and corn germ meal were replaced by nitrogen instead of soybean meal (SCCM). Cotton seed meal and corn germ meal (100%) were used as the main protein sources in three diets. Daily grain is formulated in net energy system to supplement essential amino acids in each group. After 28 days feeding, ileal contents and feces were taken as intestinal microbial analysis samples, and the microbial diversity of samples was determined by Illumina Miseq sequencing technique. The results showed that the growth performance of SBM15 and CCM was significantly lower than that of SBM15 group (P0.05), but there was no significant difference between SCCM and SBM15 group (P0.05). After the amplification and sequencing of the 16SrRNA V3-V4 region, it was found that the SCCM group day grain significantly increased the microbial diversity of the fattening pigs and increased the abundance of bacteria such as Lactobacillus and Bifidobacterium. In addition, the relative abundance of Rhomoccu, Prevoteellaceae and Bactereoides was also significantly improved in the diet group. Therefore, the soybean meal, the cottonseed meal and the corn germ meal can be used as the main protein source of the daily grain of the fattening pig, the intestinal microorganism can be adjusted, and the normal growth of the fattening pig can be maintained. A protease-producing strain 2L6 was screened from the small intestine of healthy piglets, and the strain was determined to be Bacillus cereus by morphological, biochemical and molecular biological identification. fermentation culture was carried out in soybean meal, cottonseed meal, soybean meal-cottonseed meal and fish meal fermentation medium respectively, and protease activity and substrate protein digestibility were detected at different times. The results showed that the protease activity and the substrate protein digestibility of strain 2L6 were significantly different in different fermentation medium. Among them, the protease activity in the mixed fermentation medium of soybean meal-cottonseed meal reached a maximum value of 21. 31 bb0. 09U/ m L in a relatively short time, while the digestibility of the substrate protein reached 64. 42% (P0.01) at 36h. Therefore, the mixed protein source of soybean meal-cottonseed meal can greatly improve the protease activity and the ability of protein utilization in the small intestine of piglets. in conclusion, cottonseed meal, corn germ meal and other miscellaneous meal are used as main protein source in daily grain, so that intestinal micro-organism composition of piglets and fattening pigs can be improved, that proportion of pigs can be increased, and the activity of major protease-producing strains in the intestinal tract can be improved, The method can be used as a reasonable alternative to soybean meal in the food.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S828.5
本文編號(hào)：2303296
[Abstract]:In order to explore the effect of different protein sources on the intestinal microorganisms in pigs, the effects of the diets on the microbial diversity and the major protease-producing strains of piglets and fattening pigs were studied by using cottonseed meal, fish meal and corn germ meal as the main source of protein. This study consisted of three trials. Effects of different nitrogen sources on small intestinal microorganisms in weanling piglets. Forty-eight weanling piglets (14.62. 1.46 kg) were randomly divided into 4 groups, 3 replicates in each group, 4 piglets were repeated, and the piglets were fed with single cage. The test was treated as control group (SBM group), cottonseed meal (CSM), soybean meal-cottonseed meal (SC) and fish meal (FM) group, among which, the cottonseed meal in the CSM group and SC group was 100% and 50% of the crude protein in the control group respectively. Daily grain is formulated in net energy system to supplement essential amino acids in each group. At the end of the breeding trial, 1 piglets were randomly selected for slaughter and sampling was performed using the Illumina Miseq sequencing technique to determine the diversity of the duodenum, the anterior segment, the middle segment, the posterior segment, and the ileum. The results showed that the diversity of small intestine and the main microbial composition of piglets in different protein source groups were different. Among them, Firmicutes, Proteobacteria, Bacteroidites and Acidobacter were higher in each group, and the abundance of Lactobacillus and Clostridia was higher in the two groups of cottonseed meal, while the abundance of Escherrez/ Shigella in FM group was higher. The results showed that different protein sources had a great influence on the microbial diversity of small intestine of piglets. The effects of different nitrogen sources on the growth and intestinal microorganisms of fattening pigs were studied. Thirty-six (58. 65/ 3. 71 kg) pigs were randomly divided into 4 groups, 3 replicates per process and 3 pigs per replicate. The treatment group was a control group (SBM15) group and three diets (11% CP) diet group, in which 50% of soybean meal (SBM15), cottonseed meal and corn germ meal were replaced by nitrogen instead of soybean meal (SCCM). Cotton seed meal and corn germ meal (100%) were used as the main protein sources in three diets. Daily grain is formulated in net energy system to supplement essential amino acids in each group. After 28 days feeding, ileal contents and feces were taken as intestinal microbial analysis samples, and the microbial diversity of samples was determined by Illumina Miseq sequencing technique. The results showed that the growth performance of SBM15 and CCM was significantly lower than that of SBM15 group (P0.05), but there was no significant difference between SCCM and SBM15 group (P0.05). After the amplification and sequencing of the 16SrRNA V3-V4 region, it was found that the SCCM group day grain significantly increased the microbial diversity of the fattening pigs and increased the abundance of bacteria such as Lactobacillus and Bifidobacterium. In addition, the relative abundance of Rhomoccu, Prevoteellaceae and Bactereoides was also significantly improved in the diet group. Therefore, the soybean meal, the cottonseed meal and the corn germ meal can be used as the main protein source of the daily grain of the fattening pig, the intestinal microorganism can be adjusted, and the normal growth of the fattening pig can be maintained. A protease-producing strain 2L6 was screened from the small intestine of healthy piglets, and the strain was determined to be Bacillus cereus by morphological, biochemical and molecular biological identification. fermentation culture was carried out in soybean meal, cottonseed meal, soybean meal-cottonseed meal and fish meal fermentation medium respectively, and protease activity and substrate protein digestibility were detected at different times. The results showed that the protease activity and the substrate protein digestibility of strain 2L6 were significantly different in different fermentation medium. Among them, the protease activity in the mixed fermentation medium of soybean meal-cottonseed meal reached a maximum value of 21. 31 bb0. 09U/ m L in a relatively short time, while the digestibility of the substrate protein reached 64. 42% (P0.01) at 36h. Therefore, the mixed protein source of soybean meal-cottonseed meal can greatly improve the protease activity and the ability of protein utilization in the small intestine of piglets. in conclusion, cottonseed meal, corn germ meal and other miscellaneous meal are used as main protein source in daily grain, so that intestinal micro-organism composition of piglets and fattening pigs can be improved, that proportion of pigs can be increased, and the activity of major protease-producing strains in the intestinal tract can be improved, The method can be used as a reasonable alternative to soybean meal in the food.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S828.5
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 房石山;P. Bikker;A. Dirkzwager;J. Fledderus;P. Trevisi;I. le Hueěrou-Luron;J.P.Lallès;A. Awati;;日糧蛋白質(zhì)和可發(fā)酵碳水化合物含量對(duì)剛斷奶仔豬生長(zhǎng)性能和腸道特性的影響[J];國(guó)外畜牧學(xué)(豬與禽);2007年04期
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