基于体外模拟肠道微生态系统的乳双歧杆菌MN-Gup益生元筛选及其发酵特性研究

体外肠道微生态模拟是一种研究食源性成分对肠道菌群组成和代谢产物组成的手段。本研究建立了成人的体外肠道菌群生态模拟体系,研究筛选低聚果糖、低聚半乳糖、菊粉、低聚木糖和低聚甘露糖与乳双歧杆菌MN-Gup协同增效调节肠道有益菌的作用。结果表明,添加1.5%比例低聚木糖组能够显著提升菌群中双歧杆菌的比例,而同样添加1.5%的低聚半乳糖组对乳杆菌提升更为明显,各益生元组对阿克曼菌的比例并无显著影响,菊粉组可以显著提升厚壁菌门/拟杆菌门的比值。以上结果表明,乳双歧杆菌的不同合生元组合对于肠道菌群的调控作用有明显差异,在体外肠道模拟体系下含有低聚木糖的合生元组合更有助于提高菌群中双歧杆菌的比例。     

菊粉对植物乳杆菌和保加利亚乳杆菌生长的影响

试验初步研究菊粉对植物乳杆菌和保加利亚乳杆菌生长的影响。在MRS 培养基中添加1%、2%、3%和4%的菊粉,测定2株菌生长的OD600nm位和pH。结果表 明,当菊粉添加量为1%时,促进植物乳杆菌和保加利亚乳杆菌的生长,植物乳杆菌 生长OD600nm最高为2.799,pH从5.98降到3.59;保加利亚乳杆菌生长OD600nm最高 为3.051,pH从6.55降到4.74。     

菊粉对植物乳杆菌和保加利亚乳杆菌生长的影晌

试验初步研究菊粉对植物乳杆菌和保加利亚乳杆菌生长的影响.在MRS培养基中添加1%、2%、3%和4%的菊粉,测定2株菌生长的OD600 nm位和pH.结果表明,当菊粉添加量为1%时,促进植物乳杆菌和保加利亚乳杆菌的生长,植物乳杆菌生长OD600nm最高为2.799,pH从5.98降到3.59;保加利亚乳杆菌生长OD600nm最高为3.051,pH从6.55降到4.74.     

哺乳母猪日粮中添加低聚果糖或菊粉对生产性能及血清抗体浓度的影响研究

为探讨添加0.2%低聚果糖或菊粉对哺乳母猪生产性能及血清抗体浓度的影响,选用大长母猪20头,随机分为4组,每组5头。预产期前7d开始进入产房进行预饲,分别饲喂基础日粮、基础日粮+0.2%FOS1、基础日粮+0.2%FOS2、基础日粮+0.2%菊粉,试验至产后26 d仔猪断奶时止。结果表明,0.2%FOS1可明显降低仔猪的断奶体重(P0.05),而0.2%FOS2或菊粉则对母猪生产性能无显著影响;FOS1、FOS2或菊粉能显著提高哺乳母猪及仔猪的血清IgG抗体浓度(P0.05)和提高哺乳母猪血清sIgA抗体浓度(P0.05);饲料中添加0.2%低聚果糖或菊粉能增强哺乳母猪及其仔猪的免疫功能。     

由耐热木聚糖酶从硬木和谷物木聚糖中生成低聚木糖作为短乳杆菌和青春双歧杆菌的碳源

为了比较农业生产型林业中的木聚糖,使用木聚糖RmXyn10A的两种变体,只有源自海洋红嗜热盐菌的全长酶和催化模块,对硬木(桦木)木聚糖和谷物(黑麦)阿拉伯木聚糖进行水解。用含有水解产物的低聚木糖(XOS)作为碳源,培养4种精选的细菌菌种,结果显示了短乳杆菌DSMZ 1264和青春双歧杆菌ATCC 15703出现选择性生长。这两种菌株被证实可利用低聚木糖组分(DP 2-5);而来自黑麦阿拉伯木聚糖水解产物的假定阿拉伯低聚木糖仅能被青春双歧杆菌利用。大肠杆菌不能生长,但能在单糖阿拉伯糖和木糖上生长。研究还表明,小片球菌(Pediococcus parvulus)2.6株既不能利用木糖,也不能利用低聚木糖进行生长。总之,RmXyn10A或其催化模块证实可用于硬木木聚糖和谷物阿拉伯木聚糖的高温水解,产生XOS,因此能在功能性食品中充当益生元。     

可利用表乳糖的仔猪肠道乳杆菌筛选及体外抑菌活性评价

本研究通过采用高通量测序、平板筛选、16S rDNA序列分析、特异性碳源培养和牛津杯法,从哺乳期健康仔猪的粪便中筛选分离和鉴定乳杆菌,并进行其利用表乳糖生长特性以及抑制仔猪腹泻相关致病菌活性的试验研究,探索表乳糖定向调控仔猪肠道乳杆菌增殖的可能性。结果表明:从7~10日龄哺乳期健康仔猪的粪便样品中分离出9株乳杆菌,其中唾液乳杆菌、卷曲乳杆菌、淀粉乳杆菌具有较强利用表乳糖的能力,在以2%表乳糖为唯一碳源的MRS培养基中显现出良好的生长态势,而且其培养上清液对大肠杆菌(猪源)、猪霍乱沙门氏菌、单核细胞增生李斯特氏菌、宋氏志贺氏菌以及金黄色葡萄球菌具有较好的抑菌活性。     

副猪嗜血杆菌病病原菌的分离鉴定

更有效地控制和防治副猪嗜血杆菌病,本试验从山西省晋中市疑似猪嗜血杆菌病感染的一个大型规模化猪场采集病料,选择改良巧克力琼脂培养基,从9头有关节炎、胸膜炎、腹膜炎等症状的疑似副猪嗜血杆菌病的临床病例中分离到5株疑似副猪嗜血杆菌菌株。生长特性观察发现分离株为多形性、革兰氏阴性小杆菌,生长缓慢(24～48h)。菌落呈光滑露珠样,不溶血。葡萄糖、树胶醛糖、乳糖、甘露醇、蜜三糖发酵和脲酶实验呈阴性,过氧化氢酶实验阳性,该结果符合副猪嗜血杆菌的生化特性,表明该分离菌株为副猪嗜血杆菌。     

葡萄糖及乳糖对两歧双歧杆菌生长的影响

本文研究了2种碳源——葡萄糖和乳糖对2株两歧双歧杆菌BB01及BB03生长的影响。结果表明：最适碳源因菌种而异,两歧双歧杆菌BB01的最适碳源为葡萄糖,最适添加量为1%,OD600可达1.181;两歧双歧杆菌BB03的最适碳源为乳糖,最适添加量为2%,OD600可达1.78。     

青海部分牧区牦牛酸奶中乳杆菌的分离与鉴定

利用MRS培养基,对青海部分牧区收集的15份牦牛酸奶,进行了乳杆菌的分离,对分离得到的9株疑似乳杆菌,对其中的3株进行生物学鉴定。结果:分离株均为兼性厌氧菌,在MRS培养基上,菌落较大、透明、灰白色,镜检为杆状细菌,革兰氏染色阳性,过氧化氢酶试验为阴性,乳糖发酵阳性,牛乳发酵试验阳性,符合乳杆菌的特性。     

豫陕两省14株鸡杆菌的gyrB、16S rRNA和rpoB基因序列比较分析

分析河南、陕西分离的14株鸡杆菌(Gallibacterium)之间的进化关系,及gyrB基因序列比较在该菌进化分析中的作用。PCR扩增鸡杆菌分离株的gyrB、16SrRNA和rpoB3个看家基因,PCR产物纯化后直接测序。将鸡杆菌分离株、国外参考株的3个看家基因序列进行比较分析,用Phylip 3.67软件构建进化树。结果表明,14株鸡杆菌与鸭源鸡杆菌(Gallibacterium anatis)模式株间的相似性为96.3%～98.0%(gyrB)、97.7%～99.6%(16SrRNA)和97.7%～99.0%(rpoB);14株鸡杆菌与鸡杆菌复合群1(Gallibacterium genomosp.1)参考株间的相似性为88.8%～89.9%(gyrB)、96.2%～97.5%(16SrRNA)和92.6%～93.6%(rpoB)。基于3个看家基因序列的进化分析,均显示14株鸡杆菌和鸭源鸡杆菌模式株形成单独的一个群。14株鸡杆菌分离株均属于鸭源鸡杆菌种;在3个看家基因位点,鸡杆菌河南株与陕西株之间、鸡杆菌输卵管炎病鸡分离株与健康鸡分离株之间均无明显遗传上的差异;gyrB基因序列分析可用于鸡杆菌分离株的种类鉴定,且对14株鸡杆菌与复合群1参考株的区别能力优于另外2个看家基因。     

Comparison of fermentation of selected fructooligosaccharides and other fiber substrates by canine colonic microflora

OBJECTIVE: To compare fermentation characteristics of fructooligosaccharides (FOS) and other fiber substrates that are commonly found in canine diets. SAMPLE POPULATION: Fecal samples from 3 adult dogs. PROCEDURE: The ability of fiber substrates to be used in microbial fermentation reactions was assessed by use of an in vitro fermentation system. Dogs were fed a commercially available food, and feces were collected for use as the microbial inoculum. Substrates used were beet pulp, cellulose, soy fiber, mannanoligosaccharides (MOS), FOS, and 4 inulin products (inulin 1, 2, 3, and 4). Each substrate was incubated anaerobically with fecal inoculum and growth media for 6, 12, and 24 hours, and production of short-chain fatty acids (SCFA) was measured. RESULTS: Total production of SCFA was higher for fermentation of the 4 inulin products and FOS, whereas fermentation of beet pulp, MOS, and soy fiber resulted in moderate concentrations of SCFA. Fermentation of cellulose produced the lowest concentrations of total SCFA without detection of butyrate or lactate. Butyrate production was greatest for fermentation of the 4 inulin products and FOS. Total lactate production was greatest for FOS and inulin 4. As expected, production of SCFA increased for all substrates as fermentation time increased. CONCLUSIONS AND CLINICAL RELEVANCE: Canine fecal microflora ferment FOS-containing substrates in a similar manner, with little fermentation of cellulose-based carbohydrates. Furthermore, results of an in vitro fermentation system indicate that fiber type affects the metabolic activity of microorganisms, thus influencing the amount and nature of the end products of fermentation.     

外源寡糖对奶山羊粪便微生物区系的影响

本试验旨在研究不同外源寡糖对奶山羊粪便微生物区系的影响.试验采用不完全拉丁方设计,选用平均体重为(32.80±2.45)kg、安装有永久性瘤胃瘘管的崂山奶山羊6只,分为6组,试验组羊分别饲喂在基础饲粮中添加1％的甘露寡糖(MOS组)、半乳甘露寡糖(GMOS组)、寡木糖(XOS组)、低聚异麦芽糖(IMO组)和果寡糖(FOS组)的试验饲粮,对照组(CT组)羊饲喂基础饲粮.共进行4期试验,每期35 d,其中预试期14d,正试期21 d.于正试期第1、7、14、21天晨饲前采集新鲜粪便,测定粪便中乳酸杆菌、双歧杆菌、大肠杆菌和沙门氏菌的数量.结果表明:与CT组相比,试验组乳酸杆菌数量均有所增加,其中FOS组第7天开始达到极显著水平(P＜0.01);试验组双歧杆菌数量增加,其中FOS组各采样时间均达到极显著水平(P＜0.01);试验组大肠杆菌数量有所降低,其中MOS组和FOS组第7天达到极显著水平(P＜0.01).本试验条件下,不同外源寡糖对增加奶山羊粪便乳酸杆菌、双歧杆菌数量,降低大肠杆菌数量的作用效果不同,其中果寡糖效果较明显.     

In Vitro Gas Production from Batch Cultures of Stomach and Hindgut Digesta of Horses Adapted to a Prebiotic Dose of Fructooligosaccharides and Inulin

Fructooligosaccharides (FOS) and inulin may modulate hindgut fermentation. It was tested if digesta batch cultures taken from horses adapted to FOS and inulin show different fermentation compared with such taken from nonsupplemented horses. Six horses received 0.15 g FOS and inulin/kg body weight/d via Jerusalem artichoke meal (JAM) upon a hay-based diet; six horses received corncob meal without grains (CMG) as placebo. The horses were euthanized after 20 days. Digesta samples were taken from stomach, cecum, ventral colon ascendens (VCA), and colon transversum (CT). Digesta batch cultures were incubated 48 hours to measure in vitro gas production as well as pre- and post-incubation pH and oxidation-reduction potential (ORP). A distinct fermentation of the surplus of fructans present in the inoculum was found with JAM-adapted batch cultures. Gas production was accelerated in inoculated gastric contents of horses adapted to JAM compared with CMG adapted ones (7.8 vs. 16.4 hours to achieve half of the 48 hours gas quantity, respectively; P > .05). Although buffered, pH decreased during fermentation. Postincubation pH was lower with JAM than CMG-adapted batch cultures (P > .05). Preinoculation ORP was lower with stomach batch cultures adapted to CMG than with such adapted to JAM. The ORP increased twofold from pre- to post-incubation with the latter. Asymptotic maximal gas production decreased gradually using cecum, VCA, or CT digesta. Parts of FOS and inulin of digesta are fermented in the stomach, which reduce possible effects on hindgut fermentation. Elevated fermentation may considerably impact stomach health.     

Oxalate degradation by intestinal lactic acid bacteria in dogs and cats

This study evaluated the ability of the lactic acid bacteria (LAB) component of canine and feline feces to degrade oxalate in vitro. Oxalate degradation by individual canine-origin LAB was also evaluated. The effects of various prebiotics on in vitro oxalate degradation by selected oxalate-degrading canine LAB was also evaluated. Canine fecal samples reduced oxalate levels by 78 +/- 12.2% (mean +/- S.D.; range: 44-97%, median: 81%). Feline results were similar, with oxalate reduction of 69.7 +/- 16.7% (mean +/- S.D.; range: 40-96%, median: 73%). Thirty-seven lactic acid bacteria were isolated from canine fecal samples. Mean oxalate degradation was 17.7 +/- 16.6% (mean +/- S.D.; range: 0-65%, median: 13%). No oxalate degradation was detected for four (11%) isolates, and 10/37 (27%) degraded less than 10% of oxalate. The effects of lactitol, arabinogalactan, guar gum, gum Arabic, inulin, maltodextrin or a commercial fructooligosaccharide (FOS) product on in vitro oxalate degradation by five canine LAB isolates were highly variable, even within the same bacterial species. Overall, in vitro degradation was significantly greater with guar gum compared to arabinogalactan (P < 0.05), gum Arabic (P < 0.05), and lactitol (P < 0.01). This study suggests that manipulation of the LAB component of the canine and feline gastrointestinal microflora may decrease intestinal oxalate, and correspondingly intestinal oxalate absorption and renal excretion, thus potentially reducing oxalate urolithiasis.     

不同外源寡糖对奶山羊粪便双歧杆菌数量的影响

采用平板计数法研究不同外源寡糖对崂山奶山羊粪便双歧杆菌数量的影响。试验选用安装永久性瘤胃瘘管的健康崂山奶山羊6只,采用分期分组试验设计,在不同时期分别饲喂添加1%(有效含量计)甘露寡糖、半乳甘露寡糖、寡木糖、低聚异麦芽寡糖、果寡糖的日粮,并设立空白对照组,每期35 d。结果表明,与对照组相比,试验组粪便双歧杆菌数量有所增加,其中试验第1、7、14 d果寡糖组,第21 d半乳甘露寡糖组、寡木糖组、果寡糖组均达到极显著水平(P<0.01)。本试验条件下,果寡糖对提高奶山羊粪便中双歧杆菌数量效果较明显。     

不同寡糖对肉仔鸡肠道主要菌群和免疫器官指数的影响

为研究含糖量相同(0.015%)的不同种类寡糖对肉仔鸡肠道菌群和免疫机能的影响,本试验选用150只健康的1日龄艾维茵肉仔鸡(公母各半),随机分为5个处理,每个处理设3个重复,每个重复10只鸡。对照组饲喂基础日粮,试验组为分别在基础日粮添加果寡糖组(FOS)、甘露寡糖组(MOS)、低聚异麦芽糖组(GOS)、低聚木糖组(XOS),试验期为42d。试验结果表明:42日龄时,低聚木糖组对肠道菌群的改善效果最好;寡糖极显著提高了胸腺指数(P<0.01),且低聚木糖组极显著提高了法氏囊指数(P<0.01)。     

Detection of acid production from carbohydrates by Riemerella anatipestifer and related organisms using the buffered single substrate test

One hundred and twenty-one Riemerella anatipestifer field strains from wild birds, domesticated poultry and pigs were examined for their ability to produce acid from carbohydrates by using conventional biochemical and buffered single substrate (BSS) test methods. The type strains of the species R. anatipestifer and taxometrically related genera Chryseobacterium and Bergeyella were included in the study. In contrast to 10 indole-positive R. anatipestifer variant strains, only a few of the 111 typical indole-negative R. anatipestifer strains produced acid from dextrin (32%), glucose (17%), maltose (14%) and trehalose (5%) when the conventional test procedure was used. Using the BSS test all the field isolates and the type strain of R. anatipestifer produced acid from one or more carbohydrates, most of them from dextrin (96%), maltose (91%), glucose (87%), mannose (83%), less frequently from fructose (38%) and only in some cases from trehalose (19%). One hundred and six (87%) of the R. anatipestifer strains could be assigned to 8 biovars, based on the diversity of the carbohydrate acidification patterns. The remaining 16 R. anatipestifer isolates gave delayed reactions and displayed 13 different carbohydrate acidification profiles. The Chryseobacterium and Bergeyella type strains also produced acid from more carbohydrates when the BSS test was used. The BSS-carbohydrate acidification pattern of the Chryseobacterium indologenes strain was similar to that of R. anatipestifer biovar 3.     

肉仔鸡盲肠微生物体外发酵非消化寡糖的比较研究

为了评价棉子糖、水苏糖、果寡糖、甘露寡糖等4种非消化寡糖的发酵特性,以采食无抗生素日粮的肉仔鸡盲肠微生物为菌源,通过体外发酵试验比较短链脂肪酸(SCFA)的产量及产生速率。结果表明:棉子糖发酵6 h后,SCFA产量即迅速增加,显著高于其他发酵底物(P<0.05),而且在各发酵底物产生SCFA中乙酸比例最大、丁酸比例最小(P<0.05);24 h,SCFA产量达到最大,丁酸比例在各发酵组中也增至最大。水苏糖和果寡糖的发酵速率紧随棉子糖之后,但6 h和12 h的SCFA产量仅为棉子糖的一半左右,24 h的SCFA总产量分别达到棉子糖的83.81%和75.84%。甘露寡糖的发酵速率最慢,6、12、24 h的SCFA总产量仅为棉子糖的23.81%、18.90%、40.94%。     

日粮添加水苏糖对肉仔鸡鼠伤寒沙门氏菌感染控制效果的研究

试验研究了添加不同梯度水苏糖对肉鸡鼠伤寒沙门氏菌攻毒的控制效果。试验选用1日龄体重相近(45.52±1.48)g的健康AA肉鸡公雏72只,随机分为3个处理,设6个重复,每个重复4只。在玉米-大豆浓缩蛋白日粮基础上分别添加0、0.4%和0.8%水平的水苏糖。结果表明:日粮添加水苏糖降低了肉鸡体重;肉鸡肠道沙门氏菌控制效果不显著,第19天时,0.4%水苏糖添加组的大肠杆菌数显著高于其他2组(P<0.05),乳酸菌各处理之间差异不显著;pH值与盲肠指数无差异(P>0.05)。以上研究表明,该梯度水苏糖不能控制肉鸡沙门氏菌感染,对肉鸡无保护作用。     

不同外源寡糖对崂山奶山羊瘤胃微生物区系的影响

本试验旨在研究不同外源寡糖对奶山羊瘤胃微生物区系的影响。试验选用 ６只平均体重为（３２．８０±２．４５）ｋｇ、装有永久性瘤胃瘘管的崂山奶山羊,采用分期分组试验设计。各组羊饲喂基础饲粮（对照组）以及分别在基础饲粮中添加 １％甘露寡糖（ＭＯＳ组）、半乳甘露寡糖（ＧＭＯＳ组）、寡木糖（ＸＯＳ组）、低聚异麦芽糖（ＩＭＯ组）、果寡糖（ＦＯＳ组）的试验饲粮。共进行４期试验,每期 ２３ｄ,其中预试期 １４ｄ,正试期 ９ｄ。于晨饲前（０ｈ）,晨饲后 ２、４、６、８ｈ以及晚饲前（晨饲后 １０ｈ）采集瘤胃液,测定瘤胃液中细菌总数以及纤维素分解菌、原虫和真菌数量。结果表明：与对照组相比,试验组瘤胃液细菌总数趋于增加,仅晨饲前 ＭＯＳ组显著提高（Ｐ＜０．０５）;试验组瘤胃液纤维素分解菌数量有所提高,其中 ＭＯＳ组（０ｈ）、ＧＭＯＳ组（０、２、４ｈ）、ＦＯＳ组（０、２、４ｈ）显著高于对照组（Ｐ＜０．０５）;试验组瘤胃液原虫数量有所提高,其中 ＭＯＳ组（２和 ４ｈ）、ＸＯＳ组（４ｈ）显著高于对照组（Ｐ＜０．０５）;试验组瘤胃液真菌数量有所提高,其中ＧＭＯＳ组和 ＦＯＳ组（０ｈ）极显著高于对照组（Ｐ＜０．０１）,ＭＯＳ组（２ｈ）、ＩＭＯ组（２ｈ）、ＦＯＳ组（２ｈ）、ＭＯＳ组（４ｈ）、ＦＯＳ组（６ｈ）显著高于对照组（Ｐ＜０．０５）。细菌总数、纤维素分解菌和原虫数量呈先下降后上升趋势,真菌数量呈先上升后下降趋势。本试验条件下,添加不同外源寡糖对瘤胃微生物影响有所差异,其中甘露寡糖、半乳甘露寡糖和果寡糖对瘤胃液纤维素分解菌、原虫和真菌的增殖作用较明显。