滩羊羔羊瘤胃和小肠菌群多样性的比较研究

目的 研究滩羊羔羊瘤胃和小肠菌群多样性的差异。方法 选取健康的断奶滩羊公羔,屠宰后采集瘤胃、十二指肠、空肠和回肠内容物,利用16S rDNA高通量测序技术分析瘤胃和小肠菌群结构及多样性。结果 十二指肠样品Chao1指数高于瘤胃、空肠和回肠,瘤胃样品Shannon指数和Simpson指数均高于其他部位,但差异不显著（P>0.05）。瘤胃液中拟杆菌门（Bacteroidetes）、螺旋菌门（Spirochaetae）、丝状杆菌门（Fibrobacteres）的相对丰度显著（P<0.05）高于十二指肠、空肠和回肠,十二指肠中广古菌门（Euryarchaeota）的相对丰度与瘤胃相比有升高的趋势（0.05<P<0.10）,空肠和回肠中厚壁菌门（Firmicutes）的相对丰度显著（P<0.05）高于十二指肠。瘤胃中的未鉴定菌属（Unidentified）、理研菌属_RC9（Rikenellaceae_RC9_gut_group）、密螺旋体属_2（Treponema_2）、瘤胃球菌属_NK4A214（Ruminococcaceae_NK4A214_group）、普雷沃菌属_UCG-001（Prevotellaceae_UCG-001）相对丰度显著（P<0.05）高于十二指肠、空肠和回肠,空肠中Family_ⅩⅢ_AD3011_group相对丰度显著（P<0.05）高于其他部位。结论 滩羊羔羊瘤胃中细菌的多样性更丰富,瘤胃微生物与十二指肠、空肠和回肠的细菌区系显著不同。     

Rumen microbial changes in calves fed on alpha amylase diet

Fifty-six calves in the period of a milk nutrition, randomly assigned to two groups (control and alpha amylase fed) were used to monitor the changes in the rumen microbial populations and total volatile fatty acids (VFA) concentrations associated with feeding Amylosubtilin G10X (0.6 g.day-1). Statistically significant increase was observed for amylolytic counts in experimental calves on 20th or 30th days. The counts of rumen cellulolytic bacteria and selenomonas tended to have increase in the value, but were not significantly different from those in the control valves. The results show that alpha amylase fed calves has a relative higher rumen microbial population.     

饲粮纤维水平对育肥羔羊瘤胃微生物组成及多样性的影响

试验旨在研究提高饲粮纤维水平对育肥羔羊瘤胃微生物组成及多样性的影响。试验选择健康无病、体重相近的2月龄杜湖杂交公羔,按照体重随机分为2组,每组38只,分别饲喂不同的饲粮,对照组（LW_CK）饲粮为精料+苜蓿,处理组（LW_EG）为精料+苜蓿+全株玉米青贮+花生秧;试验期150 d,其中预试期20 d,正试期130 d,试验结束后,每组随机选择5只羔羊屠宰取瘤胃内容物,通过16S rDNA高通量测序技术分析其微生物多样性及结构变化。结果表明,处理组羔羊瘤胃中细菌多样性显著高于对照组（P<0.05）。在门水平上,处理组羔羊瘤胃中优势菌门为拟杆菌门（Bacteroidetes）和厚壁菌门（Firmicutes）,对照组优势菌门为拟杆菌门和变形菌门（Proteobacteria）。提高纤维水平后显著增加了育肥羔羊瘤胃中厚壁菌门的相对丰度（P<0.05）;显著降低了变形菌门的相对丰度（P<0.05）。在属水平上,对照组与处理组羔羊瘤胃菌群共鉴定出195个属,处理组优势菌属为普雷沃氏菌属（Prevotella_1）和解琥珀酸菌属（Succiniclasticum）,对照组优势菌属为琥珀酸弧菌属（Succinivibrionaceae）和普雷沃氏菌属（Prevotella_7）。提高饲粮纤维水平,增加了育肥羔羊瘤胃中解琥珀酸弧菌属、密螺旋体属（Treponema_2）、疣微菌属（Ruminococcaceae）、月形单胞菌属（Selenomonas_1）的相对丰度,显著降低了琥珀酸弧菌属相对丰度（P<0.05）。综上所述,适量提高饲粮中纤维水平对育肥羔羊瘤胃微生物菌群结构有一定影响,能够促进部分纤维降解菌的增殖,有利于育肥羔羊的瘤胃发酵和养分消化利用。     

不同蛋白源日粮添加乳酸链球菌素对育肥湖羊瘤胃发酵及微生物菌群结构的影响

本试验旨在探究不同蛋白源日粮条件下添加乳酸链球菌素（nisin）对育肥湖羊瘤胃发酵及微生物菌群结构的影响。试验采用2×2因子设计,两因子分别为蛋白源[豆粕（SBM）、干玉米酒精糟及其可溶物（DDGS）]和nisin （添加水平为0或30.5 mg·kg^-1 DM）,两两组合,配制4种等氮等能日粮。选取32只体重（23±2） kg的断奶湖羊公羔,按照随机区组设计,根据体重分为2个区组（低体重组,16只;高体重组,16只）,每个区组的湖羊随机分配到4个组并分别饲喂对应日粮,单栏饲喂。试验预饲期1周,正式期9周,试验期结束时每组从高、低体重区组中各随机选取3只湖羊（共24只）进行屠宰,采集瘤胃内容物,提取微生物基因组DNA,利用Illumina MiSeq测序和Real-time qPCR方法分析瘤胃微生物菌群结构。研究结果表明,除Ruminococcaceae NK4A214 group和Unclassified Bacteroidetes相对丰度外,日粮蛋白源与nisin对其他所有测定指标（瘤胃发酵参数、瘤胃菌群数量、瘤胃细菌多样性及相对丰度等）均不存在交互作用（P>0.05）。日粮添加nisin对所有测定指标均无显著影响（P>0.05）。与饲喂豆粕湖羊相比,饲喂DDGS湖羊瘤胃乙酸、氨态氮浓度及总支链脂肪酸（BCVFA）浓度显著降低（P<0.05）。qPCR结果显示,饲喂不同蛋白源日粮湖羊瘤胃总菌、真菌及甲烷菌数量均无显著差异（P≥0.053）,但相对于饲喂豆粕湖羊而言,饲喂DDGS湖羊瘤胃内原虫和嗜氨梭菌（C.aminophilum）数量显著降低（P<0.05）。Illumina-MiSeq测序结果表明,饲喂DDGS湖羊瘤胃内细菌Chao1指数和ACE指数显著升高（P<0.05）。不同日粮处理组在门水平上的优势菌门均为Bacteroidetes及Firmicutes;在属水平各处理组的主要优势菌属为Prevotella 1、Christensenellaceae R-7 group和Ruminococcaceae NK4A214 group。饲喂不同蛋白源湖羊瘤胃细菌在门水平上并未产生显著影响（P≥0.14）;在属水平上,饲喂DDGS湖羊瘤胃Pseudobutyrivibrio和Roseburia丰度均显著高于饲喂豆粕湖羊（P<0.05）,而Butyrivibrio 2以及Ruminococcaceae UCG-005的相对丰度显著低于饲喂豆粕湖羊（P<0.05）。综上所述,使用DDGS作为日粮蛋白源改变了湖羊瘤胃发酵参数及微生物菌群结构,原虫和C.aminophilum数量的降低可能是导致瘤胃氨态氮浓度显著降低的主要原因;日粮添加30.5 mg·kg^-1 DM的nisin对育肥期湖羊瘤胃发酵参数及微生物菌群结构均无显著影响。     

ORIGINAL ARTICLE: Description of development of rumen ecosystem by PCR assay in milk‐fed,weaned and finished lambs in an intensive fattening system

This study examined the reticulo‐rumen characteristics of the microbial community and its fermentative characteristics in milk‐fed, at weaning and finished lambs in a conventional fattening system. Five lambs were assigned to each of three groups: milk‐fed lambs slaughtered at 30 days (T30), weaned lambs slaughtered at 45 days (T45) and ‘finished lambs’ slaughtered at 90 days (T90). At slaughter, rumen size, fermentation parameters (pH, volatile fatty acids and microbial enzyme activity) and protozoal counts were recorded. Quantitative PCR was used to quantify the genes encoding 16S and 18S ribosomal DNA of the rumen bacterial and protozoal populations, respectively, and the sequential colonization of the rumen by cellulolytic (Ruminococcus albus, Ruminococcus flavefaciens) and amylolytic (Prevotella ruminicola, Streptococcus bovis) bacteria, and protozoa (Entodinium sp.). Denaturing gradient gel electrophoresis was used to study the development of rumen microbiota biodiversity. Intake of solid food before weaning caused a significant increase in rumen weight (p < 0.0001) and bacterial DNA (p < 0.05) and volatile fatty acid analysis concentration (p < 0.01), whereas pH declined. In milk‐fed lambs, cellulolytic bacteria were evident after 30 days. Thereafter, in the 45‐day and 90‐day groups, the proportions of R. flavefaciens decreased and R. albus increased. Amylolytic bacteria were present in milk‐fed lambs; the proportion of P. ruminicola increased in fattening lambs and S. bovis was the least abundant species. Protozoal concentrations were irregular; milk‐fed lambs had a significant number of protozoa species from Entodinium and subfamily Isotrichiidae, but they disappeared at weaning. Lamb rumen were refaunated in some individuals at 90 days (Entodinium and subfamily Diplodiniinae spp.), although individual concentrations were variable.     

Observations on the alimentary tract of gnotobiotic lambs.

Seventeen gnotobiotic lambs were reared up to 21 weeks of age on cows' milk followed by sterile solid diets similar to diets fed to conventional lambs. Seven were inoculated with limited defined populations of rumen bacteria, seven were left uninoculated and three were dosed with rumen contents from conventional sheep ('conventionalised'). Seven naturally-born lambs were reared for purposes of comparison. As with other species of gnotobiotic animals, both the inoculated and the uninoculated gnotobiotic lambs had small, poorly developed lymph nodes, soft colon contents and thin intestinal walls. Unlike other species the caeca of gnotobiotic lambs were of normal size. The overall size of the reticulo-rumen including contents relative to body weight was similar in gnotobiotic and conventional lambs. However, macroscopically, the musculature of the rumen seemed to be poorly developed and histological studies showed hypoplasia of the muscle tissue of both the rumen and reticulum. Rumination was noted only infrequently in gnotobiotic lambs. The epithelium of the rumen and reticulum of the uninoculated gnotobiotic lambs was similar to that of neonatal lambs, but there was normal development of papillae in gnotobiotic lambs inoculated with limited defined populations of rumen bacteria and in conventionalised lambs. Degenerative changes were observed histologically in some of the organs of gnotobiotic lambs which were consistent with nutritional deficiencies.     

Effects of colostrum feeding and dexamethasone treatment on mRNA levels of insulin-like growth factors (IGF)-I and -II, IGF binding proteins-2 and -3, and on receptors for growth hormone, IGF-I, IGF-II, and insulin in the gastrointestinal tract of neonatal calves

The somatotropic axis regulates growth of the gastrointestinal tract (GIT). In addition, colostrum feeding and glucocorticoids affect maturation of the GIT around birth in mammals. We have measured mRNA levels of members of the somatotropic axis to test the hypothesis that colostrum intake and dexamethasone treatment affect respective gene expression in the GIT. Calves were fed either colostrum or an isoenergetic milk-based formula, and in each feeding group, half of the calves were treated with dexamethasone (DEXA; 30 microg/kg body weight per day). Individual parameters of the somatotropic axis differed (P < 0.05) among different GIT sections and formula feeding increased (P < 0.05) mRNA levels of individual parameters at various sites of the GIT. Effects of DEXA on the somatotropic axis in the GIT partly depended on different feeding. In colostrum-fed calves, DEXA decreased (P < 0.05) mRNA levels of IGF-I (esophagus, fundus, duodenum, and ileum), IGF-II (fundus), IGFBP-2 (fundus), IGFBP-3 (fundus), IGF1R (esophagus, ileum, and colon), IGF2R (fundus), GHR (fundus), and InsR (esophagus, fundus), but in formula-fed calves DEXA increased mRNA levels of IGF-I (esophagus, rumen, jejunum, and colon). Furthermore, DEXA increased (P < 0.05) mRNA levels of IGF-II (pylorus), IGFBP-3 (duodenum), IGF2R (pylorus), and GHR (ileum), but decreased mRNA levels of IGFBP-2 (ileum), and IGF1R (fundus). Whereas formula feeding had stimulating effects, effects of DEXA treatment on the gene expression of parameters of the somatotropic axis varied among GIT sites and partly depended on feeding.     

饲料来源对育肥湖羊生产性能、养分消化及瘤胃微生物组成的影响

为评价日粮粗饲料来源对育肥湖羊生产性能、养分消化及瘤胃微生物组成的影响。选择60日龄健康湖羊公羔(22.9±1.2) kg 120只,根据体重相近原则随机分成4组,每组5个重复,每个重复6只羊,分别以玉米秸秆(CS)、玉米芯(CC)、葵花籽壳(SH)和油菜秸秆(RS)作为粗饲料来源,添加比例均为20%,4组精料配比一致,预试期7 d,正试期70 d。在饲喂的49~55 d,每组选择5只羊采用全收粪法进行消化代谢试验,测定干物质(DM)、有机物质(OM)、中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)和氮(N)的表观消化率。试验结束后,每组选择15只羊进行屠宰,采集瘤胃内容物,提取瘤胃微生物DNA,对瘤胃细菌进行绝对定量。结果显示:1)CS组湖羊干物质采食量显著高于其他3组(P<0.05),且末体重和平均日增重显著高于RS组(P<0.05),其他组间差异不显著(P>0.05)。2)CS和CC组湖羊DM和ADF消化率显著高于RS组(P<0.05),且其OM和NDF消化率显著高于SH组和RS组(P<0.05);SH组湖羊N消化率显著高于RS组,其ADF消化率与其他3组湖羊差异均不显著(P>0.05)。3)CS组和CC组湖羊瘤胃中白色瘤胃球菌和普雷沃氏菌数量均显著高于RS组(P<0.05),而SH组湖羊瘤胃中两种细菌的数量与其他3组湖羊均无显著差异(P>0.05),CS组和CC组湖羊瘤胃中黄色瘤胃球菌和产琥珀酸丝状杆菌数量均显著高于SH组和RS组(P<0.05),4组湖羊瘤胃中反刍兽新月单胞菌数量差异不显著(P>0.05)。结果表明:相较于葵花籽壳组日粮和油菜秸秆组日粮,玉米秸秆组和玉米芯组日粮不仅可以确保育肥湖羊的生产性能,并且有利于纤维分解菌的生存,可促进纤维素在瘤胃中的降解。     

日粮粗蛋白质水平对育肥湖羊瘤胃微生物组成和发酵参数的影响

为研究日粮粗蛋白质水平(crude protein,CP)对育肥湖羊瘤胃微生物、pH及VFA含量的影响,选取54只4月龄[(32.25±2.50)kg]湖羊公羔,采用完全随机分组设计,将试验羊分为3个处理组,每个处理组18个重复,每个重复1只羊,3个处理组日粮CP水平分别为11.5%、14.0%、16.5%,试验期共70 d,包括7 d预试期,63 d正试期。试验期结束后,每组选择10只试验羊进行屠宰,测定屠宰性能并采集瘤胃内容物,用于瘤胃微生物DNA提取。结果表明:1)不同CP处理组对宰前活重、胴体重、屠宰率、瘤胃总重、瘤胃组织重及尾脂重无显著影响(P>0.05)。2)11.5%CP组瘤胃液氨态氮显著低于14.0%CP及16.5%CP组(P<0.001),但14.0%CP组与16.5%CP组无显著差异(P>0.05)。瘤胃液pH随日粮蛋白水平的增加有增加的趋势(P<0.1)。3)总挥发性脂肪酸(total volatile fatty acids,TVFA)浓度不受日粮中CP水平的影响(P>0.05),其中乙酸、丙酸、异丁酸、丁酸、戊酸比例均不受CP水平的影响(P>0.05),但异戊酸的比例随日粮中CP水平的增加有增加趋势(P=0.098)。4)在不同CP处理组中,瘤胃内溶纤维丁酸弧菌含量最高,但各处理组间无显著差异(P>0.05),其中白色瘤胃球菌、产琥珀酸丝状杆菌、普雷沃氏菌的含量均不受CP水平的影响(P>0.05),而黄色瘤胃球菌11.5%CP组显著低于14.0%CP组(P<0.05),16.5%CP组与11.5%CP组及14.0%CP组无显著差异。综上所述,日粮不同CP水平不影响试验羊屠宰性能,但能够影响瘤胃液pH及氨态氮含量,随CP水平的增加,异戊酸的比例有增加的趋势,14.0%CP组黄色瘤胃球菌数最多。     

不同粗精比日粮对绵羊小肠中瘤胃微生物蛋白质利用的影响

用4只装有瘤胃瘘管、十二肠瘘管和回肠瘘管的去势(公母各半)萨福克羊体重平均为(45.5±5.2)kg,在不同粗精比日粮条件下 ,研究瘤胃细菌、纤毛虫体蛋白质在小肠内的利用率。结果表明 :到达十二指肠的瘤胃细菌和纤毛虫的量 ,随日粮中精饲料水平的提高而显著减少(P<0.05)。与此相反 ,过瘤胃饲料蛋白质的量则显著增加(P<0.05)。小肠内的细菌利用率显著受日粮变化的影响 ,而纤毛虫利用率则不易受日粮影响且显著地高于细菌的利用率(P<0.05)。小肠内被吸收的细菌和纤毛虫对微生物氮的贡献率在全试验组中平均为50 %。     

Localization of bacterial antigens in calves inoculated orally with ruminal Bacteroides succinogenes and Selenomonas ruminantium

Localization of bacteria and bacterial materials was investigated in calves inoculated orally with live organisms of both Bacteroides succinogenes and Selenomonas ruminantium by a immunohistological method using rabbit antiserum against the outer membrane of those organisms and by a scanning electron microscope. The intact organisms of both inoculated bacterial species were detected on the rumen wall and in the lamina propria of the forestomach, and S. ruminantium also in the lymph nodes associated with the rumen. The bacterial materials were observed inside of macrophage-like cells in the lamina propria of the forestomach and of lymphoid cells in the lymph nodes. Number of the cells in the mesenteric lymph nodes was smaller than that of the forestomach associated lymph nodes. The results suggest that the orally inoculated bacteria may act as antigenical stimulant in the mucosa of the forestomach and associated lymph nodes of calves.     

Effects of protozoa on methane production in rumen and hindgut of calves around time of weaning.

Effects of the presence or absence of ciliate protozoa on methanogenesis in the rumen and hindgut were investigated in young calves during a 7-week period. Ten Holstein calves, aged 7 days, were divided in two groups (n = 5) and fed an increasing amount of a commercial milk replacer and small amounts of a calves starter. One group was inoculated with ciliate fauna on two occasions, week 5 and 6, while the second remained ciliate-free. The absence of protozoa in the rumen decreased rumen empty weight (-23%, P < 0.01), and rumen pool size of N (-36%, P < 0.01) and crude fat (-37%, P < 0.05). Rumen bacteria of non-faunated calves contained a higher proportion of total amino acid-N per 16 g N (+3%, P < 0.01) and D-alanine-N per 16 g N (+13%, P < 0.05) compared to faunated calves. Further results contain a reference for a higher bacterial mass in the ciliate-free rumen with an increased number of bacteria adherent to rumen mucosa. The CH4 production in the rumen increased exponentially with the increase in protozoa population size (R2 = 0.68). In presence of 46 x 10(4) protozoa per ml rumen fluid, the in vitro CH4 production of rumen fluid per mol total VFA was about 34% higher in faunated than in non-faunated calves (P < 0.001). Hydrogen (2H) recovery of rumen fermentation was positively correlated (R2 = 0.55) to the CH4 production rate. Methanogens were attached on rumen mucosa. Methanogenesis, induced by rumen mucosa attached bacteria, was stimulated by ruminal protozoa. In the absence of protozoa in the rumen, the acetate-propionate ratio and butyrate proportion of VFA were reduced. In vivo, in the absence of protozoa not only the whole animal CH4 production (-30%, P < 0.05) but also the digestibility of carbohydrates (-4%, P < 0.05) was reduced. Thereby no difference was observed in the intake of ME per kg DM between the groups. In conclusion, the methanogenesis in the rumen, but not in hindgut, is associated with the development of the ruminal protozoa population. The level of methanogenesis (mol/mol VFA) in the hindgut amounts to 20% of the ruminal methanogenesis.     

饲养水平对阿勒泰羊胃肠道发育、瘤胃发酵参数及瘤胃微生物区系的影响

为研究饲养水平对阿勒泰羊胃肠道发育、瘤胃发酵参数、消化酶活性及瘤胃微生物区系的影响,选取月龄相近（3~3.5月龄）、体重均一（19.16±0.54） kg、臀型一致、健康状况良好的阿勒泰母羔羊30只,随机分为3组,每组10只,自由采食60 d后,参照NRC（2007）饲养标准中维持能量（Em）需要,3个组分别按1.5Em（0.75 MJ·kg^-1 W^0.75）、1.0Em（0.50 MJ·kg^-1 W^0.75）和0.5Em（0.25 MJ·kg^-1 W^0.75）饲喂30 d。结果表明：1）随饲养水平的降低,羔羊全胃、瘤胃、小肠的重量逐渐降低,且1.5Em组全胃、瘤胃、小肠重量和瘤胃指数显著高于1.0Em和0.5Em组（P<0.05）,1.5Em和1.0Em组小肠长度和小肠指数显著高于0.5Em组（P<0.05）,但1.5Em和1.0Em组无显著差异（P>0.05）;2）随饲养水平的降低,绵羊瘤胃液pH先升高后降低,1.5Em组pH显著低于0.5Em和1.0Em组（P<0.05）,氨态氮（NH₃-N）含量逐渐降低但3组之间的差异不显著（P>0.05）,总挥发性脂肪酸（TVFA）以及乙酸、丙酸、丁酸、戊酸、异丁酸、异戊酸含量1.0Em组最低,且1.0Em组TVFA、乙酸、丙酸含量显著低于1.5Em和0.5Em组（P<0.05）,丁酸、戊酸和乙酸/丙酸1.0Em和0.5Em组显著低于1.5Em组（P<0.05）;3）随饲养水平的降低,瘤胃液消化酶活性降低,脂肪酶、蛋白酶活性1.5Em组显著高于1.0Em和0.5Em组（P<0.05）,1.0Em和0.5Em组之间差异不显著（P>0.05）,α-淀粉酶3组之间差异显著（P<0.05）;4）瘤胃内容物中真菌、原虫的相对含量1.5Em组显著高于1.0Em和0.5Em组（P<0.05）,但1.0Em和0.5Em组之间差异不显著（P>0.05）,产琥珀酸丝状杆菌、白色瘤胃球菌1.0Em组最低,显著低于1.5Em和0.5Em组（P<0.05）,黄色瘤胃球菌3组之间无显著变化（P>0.05）。由此可见,饲养水平对阿勒泰羊胃肠道发育、瘤胃发酵参数、消化酶活性及瘤胃微生物区系都具有显著影响。     

宏基因组测序分析野生林麝瘤胃、小肠和大肠微生物组成和抗生素抗性基因

本试验旨在研究野生林麝(Moschus berezovskii)瘤胃、小肠和大肠微生物组成和抗生素抗性基因.采集野生林麝消化道3个区段(瘤胃、小肠和大肠)的内容物进行宏基因组测序,并进行常规物种注释和抗生素抗性基因功能注释.结果表明:3个区段共有优势菌门为厚壁菌门(Firmicutes)和变形菌门(Proteobact...     

Glutamate dehydrogenase and glutamine synthetase activity in the digestive tract in lambs in relation to age]

Developmental dynamics was investigated in the activity of glutamate dehydrogenase (GDH, E.C. 1.4.1.2.-4) and glutamine synthetase (GS, E.C. 6.3.1.2) in different parts of the digestive tract of lambs, in dependence on the age from 10 to 90 days; the goal of these investigations was to elucidate in greater detail the role of the above enzymes in nitrogen metabolism. The activity of GDH, and of the coenzymes NADH and NADPH, was followed in the digesta because simple organisms (bacteria, fungi, plants) have two glutamate dehydrogenases: they differ from each other by coenzyme specificity, unlike GDH from animal sources which can utilize both NADH coenzyme and NADPH coenzyme (Fahien et al., 1965; Frieden, 1964). The following activities of GDH and GS were found out in trials with lambs at the age of 10, 20, 30, 40 and 90 days, as to the different parts of digestive tract: in the tissues of rumen, omasum, reticulum, spleen, duodenum, jejunum, ileum, int. caecum and colon the activity of GDH (NADH) varied from 0.031 to 0.305 nkat/mg dry matter, in the digesta from 0 to 2.92 nkat/mg dry matter. An investigation of GDH (NADH, NADPH) dynamics in the digesta of lambs showed the relatively high activity of GDH (NADH) in the digesta of colon at the age of 10 days and that of GDH (NADPH) in the digesta of int. caecum. The activity of GDH (NADH) was also found to be high in the digesta of int. caecum at the age of 20 days. In that period the activity of GDH (NADH, NADPH) in the digesta of rumen, omasum and reticulum was zero.(ABSTRACT TRUNCATED AT 250 WORDS)     

Study of the effect of presence or absence of protozoa on rumen fermentation and microbial protein contribution to the chyme

The aim of this study was to investigate the effect of presence or absence of protozoa on rumen fermentation and efficiency of microbial protein synthesis under different diets. Of 20 twin paired lambs, 1 lamb of each pair was isolated from the ewe within 24 h after birth and reared in a protozoa-free environment (n = 10), whereas their respective twin-siblings remained with the ewe (faunated, n = 10). When lambs reached 6 mo of age, 5 animals of each group were randomly allocated to 1 of 2 experimental diets consisting of either alfalfa hay as the sole diet, or 50:50 mixed with ground barley grain according to a 2 × 2 factorial arrangement of treatments. After 15 d of adaptation to the diet, the animals were euthanized and total rumen and abomasal contents were sampled to estimate rumen microbial synthesis using C(31) alkane as flow marker. Different ((15)N and purine bases) and a novel (recombinant DNA sequences) microbial markers, combined with several microbial reference extracts (rumen protozoa, liquid and solid associated bacteria) were evaluated. Absence of rumen protozoa modified the rumen fermentation pattern and decreased total tract OM and NDF digestibility in 2.0 and 5.1 percentage points, respectively. The effect of defaunation on microbial N flow was weak, however, and was dependent on the microbial marker and microbial reference extract considered. Faunated lambs fed with mixed diet showed the greatest rumen protozoal concentration and the least efficient microbial protein synthesis (29% less than the other treatments), whereas protozoa-free lambs fed with mixed diet presented the smallest ammonia concentration and 34% greater efficiency of N utilization than the other treatments. Although (15)N gave the most precise estimates of microbial synthesis, the use of recombinant DNA sequences represents an alternative that allows separate quantification of the bacteria and protozoa contributions. This marker showed that presence of protozoa decrease the bacterial-N flow through the abomasum by 33%, whereas the protozoa-N contribution to the microbial N flow increased from 1.9 to 14.1% when barley grain was added to the alfalfa hay. Absolute data related to intestinal flow must be treated with caution because the limitations of the sampling and maker system employed.     

热带假丝酵母菌与桑叶黄酮对犊牛营养物质代谢和瘤胃发酵的影响

本文旨在研究饲粮中添加热带假丝酵母菌(CT)与桑叶黄酮(MLF)对断奶前后犊牛能量与氮代谢、瘤胃微生物蛋白合成以及瘤胃发酵的影响。选取(20±2)日龄中国荷斯坦公犊牛48头,随机分为4组,对照组(CON)饲喂基础饲粮,断奶前为代乳品和开食料,断奶后为开食料;CT组在基础饲粮中添加CT;MLF组添加MLF;CM组添加CT和MLF。犊牛于56日龄断奶(代乳品),试验期60 d。分别于犊牛28、42、56和80日龄采集瘤胃液,35和63日龄进行消化代谢试验。结果表明:与CT组相比,CM组犊牛断奶前代谢能显著提高(P0.05),较CON组,断奶后CM组氮利用率显著提高(P0.05);断奶前MLF组较CON和CT组氮的生物学价值显著提高(P0.05)。与CON组相比,CT组显著提高了28~80日龄瘤胃液p H和56、80日龄瘤胃液微生物蛋白含量(P0.05),CT、MLF和CM组瘤胃液氨态氮浓度无显著变化(P0.05);与CON组相比,56日龄M LF组总挥发性脂肪酸浓度和丁酸含量显著提高(P0.05);42日龄乙酸/丙酸CT和MLF组较CON组有提高的趋势(P=0.090),各组间乙酸和戊酸含量无显著差异(P0.05)。结果提示,饲粮添加CT有助于改善断奶前犊牛瘤胃液p H和促进瘤胃微生物蛋白的合成;添加CT和MLF有助于提高断奶前犊牛饲粮代谢能和氮的生物学价值,提高断奶后犊牛总能代谢率和氮的利用率,降低粪能和总排出氮量,同时具有改善瘤胃发酵的作用,并且CT与MLF联合使用效果优于单一添加。     

羔羊消化器官的早期生长发育和瘤胃内主要消化酶活性的变化

选用健康无病的波尔山羊30只,饲养在相同条件下,分别于20日龄和60日龄,各选取体重相近的3只羔羊屠宰,研究羔羊消化器官的生长发育和瘤胃内主要消化酶活性的变化规律。结果表明:两个日龄之间,消化道各部位的重量差异显著(P<0.05);复胃或瘤胃重量占体重的比例差异极显著(P<0.01),小肠或大肠的重量占体重的比例差异显著(P<0.05);瘤胃、十二指肠、盲肠中的pH60日龄显著(P<0.05)高于20日龄;回肠pH60日龄高于20日龄,但差异不显著(P>0.05);皱胃pH60日龄低于20日龄;瘤胃液中淀粉酶和木聚糖酶的活性增长迅速,60日龄显著(P<0.05)高于20日龄,而纤维素酶和果胶酶虽有一定程度的增长,但差异不显著(P>0.05)。     

DAP-decarboxylase activity and lysine production by rumen bacteria.

The last step of pathway of lysine biosynthesis by rumen bacteria was tested. The first measurements of DAP-decarboxylase activity and of lysine production by Megasphera elsdenii, Selenomonas ruminantium, Clostridium spp., Butyrivibrio fibrisolvens and Bacteroides succinogenes as well as the first attempts to increase the lysine production by ruminal streptococci by mutation are described. The highest values were measured in Selenomonas ruminantium (DAP-decarboxylase activity = 146 micrograms DAP.min-1.mg-1 protein and lysine production was 390 micrograms.mg-1 protein) and the lowest values were ascertained in Butyrivibrio fibrisolvens (DAP-decarboxylase activity = 27 micrograms DAP.min-1.mg-1 protein and lysine production was 32 micrograms.mg-1 protein). DAP-decarboxylase activity was increased by mutation especially in Streptococcus bovis, the lysine production in both of tested ruminal streptococci. The potential use of lysine-excreting mutants in calves in future is suggested.     

甘露寡糖添加方式对哺乳期犊牛瘤胃细菌菌群结构的影响

为利用16S rDNA技术研究甘露寡糖不同添加方式对哺乳期犊牛瘤胃细菌菌群结构影响,本研究选用出生日龄一致、体重接近、健康状况良好的荷斯坦公犊牛20头,随机分为4组。CR组为对照组,饮用乳及开食料中均不添加甘露寡糖;ORa组在饮用乳中添加5 g甘露寡糖,开食料中不添加甘露寡糖;ORb组在开食料中添加5 g甘露寡糖,饮用乳中不添加甘露寡糖;ORc组在饮用乳及开食料中各添加2.5 g甘露寡糖(混合添加)。应用16S rDNA测序技术,研究了甘露寡糖添加方式对犊牛瘤胃细菌构建的影响。结果表明:甘露寡糖添加方式会影响哺乳期犊牛瘤胃细菌总数,但对哺乳期犊牛瘤胃细菌菌群多样性未产生显著影响(P>0.05)。在门水平上,与对照组相比,ORb组颗粒料中添加甘露寡糖显著降低了厚壁菌门与放线菌门的丰度(P<0.05),极显著提高了变形菌门的丰度(P<0.01),优势菌门增加为3种,分别为拟杆菌门、厚壁菌门与变形菌门。在属水平上,甘露寡糖不同添加方式导致哺乳期犊牛瘤胃细菌菌群结发生了改变,其中甘露寡糖不同添加方式对小杆菌属和脱硫弧菌属在瘤胃内丰度均产生了影响,但对ORa组与ORc组犊牛瘤胃细菌影响不及ORb组;且ORb组犊牛瘤胃细菌功能性菌属普雷沃氏-7属、氨基酸球菌属、优杆菌属和琥珀酸弧菌等均发生显著变化。基于以上结果,得出如下结论:本研究条件下,甘露寡糖添加方式对哺乳期犊牛瘤胃细菌菌群多样性未产生显著影响(P>0.05),但开食料中寡糖添加方式对菌群结构产生部分影响,其中发挥瘤胃蛋白降解作用和淀粉降解作用的琥珀酸弧菌科-UCG-001属、利用乳酸的新月形单胞菌属丰度极显著提高(P<0.01),而利用乳糖的优杆菌属丰度显著降低,降解半纤维素的小杆菌属丰度极显著降低(P<0.01)。