DNA of some anaerobic rumen fungi: G + C content determination

The nuclear DNAs from five species of anaerobic rumen fungi have been isolated and purified by means of two extraction methods (with and without 8 M urea). Their G + C contents have been characterized by the thermal denaturation procedure of Marmur and Doty. As has already been shown in Neocallimastix frontalis, the results obtained by the two techniques demonstrated a very low G + C content (less than 20%) and the constant presence of satellite DNA.     

The effects of concentrate energy source on feed intake and rumen fermentation parameters of dairy cows offered a range of grass silages

The effects of concentrate energy source on feed intake and rumen fermentation parameters of lactating dairy cattle, offered one of three grass silages differing in fermentation and intake characteristics, were evaluated in a partially balanced changeover design experiment involving four rumen fistulated dairy cows. Three silages were harvested using different management practices prior to and at ensiling. Silages A and C and silage B were harvested from primary or secondary regrowths either untreated or treated with a bacterial inoculant. For silages A, B and C, dry matter (DM) concentrations were 334, 197 and 183 g/kg (S.E. 3.1), pH values 4.00, 4.79 and 4.80 (S.E. 0.042) and ammonia nitrogen (N) concentrations were 123, 319 and 295 g/kg total N (S.E. 20.0), respectively. Two concentrates were formulated to contain similar crude protein, effective rumen degradable protein, digestible undegradable protein and metabolisable energy concentrations but using different carbohydrate sources to achieve a wide range of starch concentrations. For the low and high starch concentrates starch concentrations were 17 and 304 g/kg DM and acid detergent fibre concentrations were 170 and 80 g/kg DM, respectively. The silages were offered ad libitum, supplemented with 10 kg fresh concentrate daily. For silages A, B and C, DM intakes were 10.9, 7.2 and 8.6 kg/day and concentrate energy sources did not alter (P>0.05) intake. Increasing the level of starch in the concentrate decreased the molar concentration of acetate (P<0.05) and tended to increase the molar concentration of propionate (P<0.1). Silage type altered the molar concentration of acetate (P<0.01) and the acetate:propionate ratio (P<0.05). There were no silagetype×concentrate interactions (P>0.05) on silage intake or rumen fermentation parameters. It is concluded that when concentrate and silage form equal proportions of the diet, the composition of the silage has an over-riding influence on rumen fermentation parameters. Furthermore, the changes in milk fat concentration, observed in a concurrent production study, due to changes in silage and concentrate types can be accounted for by changes in the ratio of lipogenic to glucogenic precursors in the rumen fluid.     

Anaeromyces mucronatus nov. gen., nov. sp. A new strictly anaerobic rumen fungus with polycentric thallus

A new species of strictly anaerobic fungus was isolated from the cow rumen. It is characterized by a polycentric thallus, a polynuclear rhizomycelium, mucronate zoosporangia and uniflagellated zoospores. It is also singular in that the sporocysts do not react to the specific lectins of L-fucose, N-acetyl-D-galactosamine and diacetyl chitobiose. These characteristics justify the creation of a new genus.     

In vitro fibrolytic potential of anaerobic rumen fungi from ruminants and non-ruminant herbivores

In the present study, anaerobic fungi were isolated from different ruminants and non-ruminants; i.e., cattle, buffalo, sheep, goats, wild bluebulls, elephants, deer, and zebras; and were identified as Anaeromyces, Orpinomyces, Caecomyces, Piromyces, and Neocallimastix sp., based on their morphological characteristics. These isolates possessed significant in vitro hydrolytic enzyme activities; however, an isolate of Caecomyces sp. from elephant was found to exhibit maximum activity, i.e., filter paper cellulase (Fpase; 21.4 mIU/ml), carboxymethyl cellulose (CMCase; 15.1 mIU/ml), cellobiase (37.4 mIU/ml), and xylanase (26.0 mIU/ml). Besides, this isolate also showed the significantly highest ability to digest plant cell-wall contents in vitro. The in vitro dry matter digestibility increased from 45.1 to 48.9% after 48 h of incubation, and the plant cell-wall contents, in terms of neutral detergent fiber and acid detergent fiber, decreased from 64.2 to 61.3% and from 31.3 to 29.6%, respectively. These results indicate that such fibrolytic ruminal fungal strains are prevalent in wild herbivores such as elephants, as well as in other ruminants and non-ruminants, and could be exploited as microbial feed additives for improved nutrition and productivity in domesticated ruminants.     

利用基本培养基初步筛选牛瘤胃中纤维素降解菌

目的初步筛选牛瘤胃中纤维素降解菌。方法分别采用基本培养基（牛肉膏蛋白胨培养基、马丁培养基）,利用好氧、兼性和厌氧3种不同的培养方法进行初选,初步分离牛瘤胃中的细菌与真菌,再通过复选培养基（加入微晶纤维素）,筛选降解纤维素的菌种。结果筛选分离出降解纤维素的1株厌氧细菌和1株厌氧真菌。结论此实验方法简单易行,能够有效地从牛瘤胃中筛选出生长良好的纤维素降解菌。     

Comparison of protein fermentation characteristics in rumen fluid determined with the gas production technique and the nylon bag technique

In this study, a modified version of the gas production technique was used to determine protein fermentation characteristics in rumen fluid of 19 feedstuffs. Performing the incubations in a N-free environment, and with an excess of rapidly fermentable carbohydrates, made N the limiting factor to microbial growth, and so gas production profiles reflected the availability of N from the feed samples. Results showed that fermentation of protein in rumen fluid can be determined with this modified gas production technique, and that there were distinct differences in protein fermentation between the feed samples. Availability of protein for fermentation was highest in wheat, potato pieces and lupin, and lowest in Rumiraap, a formaldehyde treated rapeseed meal, palm kernel expeller and brewery grains. The protein degradation characteristics of the 19 feed ingredients were also determined with the in situ nylon bag technique. With the obtained results, the amount of rumen escape protein (REP) was calculated for each feedstuff. The results showed that the rate of degradation ranged from 0.010/h for Rumiraap to 0.151/h for wheat. The amount of REP ranged from 197 g/kg CP for lupin to 840 g/kg CP for Rumiraap. Comparing the gas production results with the results obtained with the nylon bag technique showed that there was a good relationship between the gas production after 12–25 h of incubation and the calculated amount of REP (r² = 0.83–0.85). The results show that the adapted gas production technique, being depleted of N and using an excess of rapidly fermentable carbohydrates, is suitable to recognize differences in N availability between feed samples and can be used as an alternative to the nylon bag technique and other in vitro techniques.     

Effects of isobutyrate on rumen fermentation, lactation performance and plasma characteristics in dairy cows

The objective of this study was to evaluate effects of isobutyrate supplementation on rumen fermentation, lactation performance and plasma characteristics of dairy cows. Twenty multiparous second filial generation (F2) cows of a cross between Chinese Jinnan Yellow and Holstein cows at 148 ± 4.5 days in milk and 22.3 ± 0.81 kg milk production were used in a replicated 4 × 4 Latin square experiment. The treatments were: control (without isobutyrate), low (LIB), medium (MIB) and high (HIB) isobutyrate supplementation of 20, 40 and 60 g per cow per day, respectively. Experimental periods were 30 days with 15 d of adaptation and 15 d of data collection. Dry matter (DM) intake was not affected by increasing isobutyrate supplementation, but milk yields were highest for the 40 g/d isobutyrate supplementation level, where proportion of milk fat, true protein and lactose were minimized. Ruminal pH (6.38–6.24) and ammonia N (13.8–11.1 mg/100 ml) were linearly (P<0.01) decreased, whereas total VFA concentration (124–131 mM) increased at a decreasing rate with increasing isobutyrate supplementation. The ratio of acetate to propionate increased linearly (P<0.01) from 2.77 to 4.43 as isobutyrate supplementation increased due to the increase in acetate production and decrease in propionate production. Digestibilities of OM in the total tract increased linearly (P<0.01) as isobutyrate supplementation increased, digestibilities of DM and EE were highest for the 40 g/d isobutyrate supplementation level, digestibilities of CP, aNDF and ADF increased at a decreasing rate with increasing isobutyrate supplementation. Plasma concentrations of glucose and growth hormone linearly (P<0.03) increased, whereas concentrations of non-esterified fatty acids linearly (P<0.01) decreased. Results indicate that supplementation of this diet with isobutyrate changed the rumen fermentation pattern towards acetate production, improved digestion and modified plasma concentrations of glucose and growth hormone. This suggests that isobutyrate stimulated digestive microorganisms or enzymes in a dose-dependent manner with the optimum isobutyrate dose at about 40 g per cow per day in terms of improved digestion.     

Influence of exogenous enzymes on nutrient digestibility, extent of ruminal fermentation as well as milk production and composition in dairy cows

This experiment studied effects of a mixture of exogenous enzymes (ZADO^®) from anaerobic bacteria on ruminal fermentation, feed intake, digestibility, as well as milk production and composition in cows fed total mixed rations (TMRs; 0.7 corn silage and 0.3 of a concentrate mixture). Twenty lactating multiparous Brown Swiss cows (500 ± 12.4 kg live weight) were randomly assigned into two experimental groups of 10 immediately after calving and fed a TMR with or without (CTRL) addition of 40 g/cow/d of enzymes for 12 weeks. Addition of enzymes increased (P<0.05) rumen microbial N synthesis. Intake of dry matter (DM) and organic matter (OM) was positively influenced (P<0.05) by supplementation, and digestibility of all nutrients was higher (P<0.05) in the total tract of supplemented cows, although the magnitude of the improvement varied among nutrients, with the highest improvement in aNDFom and ADFom (418–584 and 401–532 g/kg respectively; P<0.05) than the other nutrients. Supplementation of enzymes also increased (P<0.05) rumen ammonia N and total short chain fatty acid (SCFA) concentrations, and individual SCFA proportions were also altered with an increase in acetate (61.0–64.8 mol/100 mol; P=0.05) before feeding, and acetate and propionate increased 3 h post-feeding (60.0–64.0 and 18.3–20.8 mol/100 mol respectively; P<0.05). Milk and milk protein production was higher (12.8–15.7 and 0.45–0.57 kg/d respectively; P<0.05) for cows fed the ZADO^® supplemented diet. This exogenous enzyme product, supplemented daily to the TMR of cows in early lactation, increased milk production due to positive effects on nutrient intake and digestibility, extent of ruminal fermentation and microbial protein synthesis.     

广西水牛瘤胃中的细菌多样性

[目的]了解广西水牛瘤胃中细菌的组成及其可能的降解纤维素细菌的主要类群。[方法]提取水牛瘤胃内容物和高效降解滤纸的水牛瘤胃内容物的富集培养物的宏基因组DNA,以宏基因组DNA为模板,扩增16S rRNA基因序列,构建该两种样品的细菌的16S rRNA基因文库。通过对16S rRNA基因序列的分析,了解这两种样品的细菌群体种类及数量。 [结果] 水牛瘤胃内容物与其富集培养物中均主要含有LGCGPB (low G+C Gram-Positive Bacteria)、CFB (Cytophaga-Flexibacter-Bacteroides)两大类菌群和少数的螺旋体菌（Spirochaetes）,且LGCGPB所占的比例都是最高的,LGCGPB在水牛瘤胃内容物细菌中的比例为56.66%,而在富集培养物细菌中的比例升高为73.33%。在水牛瘤胃内容物中,丝状杆菌（Fibrobacteres）占3.33%,但在富集培养物中未被检测到。而在富集培养物中占13.33%的变形杆菌（Proteobacteria）,在水牛瘤胃内容物中未被检测到。本研究还发现了分类地位尚未明确的一菌群（R46）。[结论]细菌类群LGCGPB、Proteobacteria可能在水牛瘤胃中的纤维素降解过程中起重要作用。此外,水牛瘤胃中的细菌组成和牦牛、牛、羊瘤胃中的细菌组成较相似但比例有所不同。