Influence of kanamycin on the rumen bacteria and ammonia production in supplemented sheep rumen liquor in vitro

Bacteriological studies were made with in vitro sheep ruminal fluids supplemented with synthetic fodder containing different amounts of kanamycin (0.125, 2.50, and 60 mg/ml), during anaerobic incubation at 38–39°C for 6 h.Changes in the bacterial population occurred in the presence of large amounts (60 mg/ml) of kanamycin, and the B₁-type strain of Eubacterium became predominant.The marked increase in the concentration of ammonia in the supplemented rumen liquor, obtained after anaerobic incubation with high dosages of this antibiotic, is discussed.     

绵羊粪与瘤胃液作菌源对纤维体外消化的比较研究

本研究以绵羊粪和瘤胃液作菌源,测定了4类(豆科、禾本科、秸秆及青贮)、9种粗饲料及5个混合粗饲料组中的中性洗涤纤维(NDF)的体外消化率(IVNDFD),用两种菌源的可发酵纤维指数(FFI)对这些粗饲料的纤维品质进行了评定。按两种菌源的FFI的大小对所测定的粗饲料纤维品质进行的排序完全一致:青干草>玉米青贮>玉米秸>羊草>草地早熟禾>谷草>混粗C2>混粗C1>混粗D>苜蓿>沙打旺>苏丹草;并以两种菌源不能消化的NDF(unNDF)为预测因子,建立了预测绵羊干物质随意采食量(DMI)的模型,比较、分析发现两种模型等效。考虑到不需使用瘘管动物,建议在生产实践中使用以绵羊粪作菌源所不能消化的NDF(unNDF)为预测因子的DMI模型:DMI豆(g/d·kgw0。75)=43.43/unNDF(％DM);DMI禾(g/d·kgw0.75)=33.89/unNDF(％DM);DMI秸(g/d·kgw0.75)=23.84/unNDF(％DM):DMI贮(g/d·kgw0.75)=22.42/unNDF(％DM)。     

Extracorporeal perfusion of the sheep rumen]

We constructed a modified perfusion apparatus and elaborated a method of extracorporal perfusion of the rumen of sheep. As perfusates we used the bovine plasma diluted in a ratio of 1:1 of an isotonic sodium chloride (NaCl) solution and the whole autologous blood. Transaminases GOT and GPT, ammonia and pH were determined in the perfusate. The different perfusions were evaluated according to previously determined perfusion conditions and criteria. A subject for discussion is the question of suitability of the parameters under examination for judging the state of the perfused organ. The described method is suitable for the study of metabolical processes in the rumen wal.     

Factors that limit maintenance of recombinant rumen bacterium in sheep rumen

Factors limiting the maintenance of recombinant ruminal bacterium in the rumen were evaluated in vitro , using batch culture prepared from rumen fluid of sheep. Butyrivibrio fibrisolvens expressing a foreign xylanase gene ( B. fibrisolvens NO4) was used as a tested recombinant that was selectable on an erythromycin-containing agar medium. The recombinant tended to reduce its level slowly in the rumen fluid of sheep on a high hay diet, while its initial decrease was more apparent in the rumen fluid of sheep on a high concentrate diet. Incubation with cell-free ruminal fluid revealed a significant decrease of inoculated recombinant, suggesting the presence of antibacterial factors limiting maintenance of the recombinant. In particular, during the first 12 h of incubation this inhibition was more notable in culture prepared from rumen fluid of sheep given the high concentrate diet. Autoclaving the cell-free rumen fluid inactivated the inhibition. Numbers of the recombinant for inoculation did not influence the final level of survived recombinant, that is, the initial depression was larger as more recombinant was inoculated. Subculturing with xylan before inoculation and/or direct addition of xylan to the batch culture did not improve survival of the recombinant. From these results it is suggested that the level of survived recombinant is limited to 10^2–4/mL of in vitro batch culture with restricted energy supply and that initial depression of the recombinant is mainly caused by the heat-sensitive antibacterial factors not associating with microbial cells in the rumen.     

Stability of ivermectin in rumen fluids

To determine whether ivermectin is metabolized in the rumen, in vitro studies were conducted with the tritium-labelled H₂B_1a component of ivermectin in rumen fluid from sheep and cattle. No detectable metabolism occurred over 24 h in in vitro incubations at 38°. The viability of the microbes in the rumen fluids was demonstrated by the conversion of 17% and 11% of [¹⁴C]cellulose to ¹⁴CO₂ in 24 h in the incubations with sheep and steer rumen fluids respectively. The results indicate that ivermectin is not metabolized in the rumen. Based on the lack of in vitro metabolism of ivermectin in rumen fluid, the similarity of in vitro liver microsomal metabolism with in vivo metabolism of the avermectins and the physicochemical properties of the avermectins, any disappearance of ivermectin in vitro from rumen fluid is probably a result of binding to solids or surfaces. Apparent discrimination by dung beetles, where observed, between control faeces and faeces from cattle or sheep treated with ivermectin or abamectin therefore must be attributable to chance, to factors unrelated to treatment or to factors such as changes in amino acid composition rather than the production of volatile metabolites of ivermectin.     

Disappearance of ethanol from isolated sheep rumen

The absorption of ethanol from the rumen was studied in three British Milk sheep equipped with a rumen cannula. After removal of the rumen content and washing the forestomachs several times the reticulo-omasal orifice was closed and through the cannula 20 or 60 ml ethanol and 2 ml Cr-EDTA were infused in physiological saline. The entire fluid volume was 3000 ml. At the start of the experiment (0 min) and subsequently in the 5th, 15th, 30th, 45th, 60th and 75th minutes samples were taken from the fluid present in the forestomachs. During the 75-min experiment the amount of ethanol gradually decreased in the rumen. The rate of disappearance varied according to concentration. The graph depicting the change of ruminal ethanol concentration shows a curve typical of passive transport. The equation describing the disappearance of ethanol was y = -0.0474x2 + 5.6544x + 10.869 after the administration of 20 ml ethanol, and y = -0.1377x2 + 19.541x - 24.606 after the infusion of 60 ml ethanol. It was established that ethanol was absorbed through the rumen wall by a passive transport process.     

Effect of defaunation and refaunation of the rumen on rumen fermentation and N-flow in the duodenum of sheep

In order to confirm earlier fragmentary results, the effect of defaunation and refaunation of the rumen on the fermentation pattern and flow of N-components in the proximal duodenum of two sheep was investigated. Defaunation had no effect on acetic acid as a proportion of the total volatile fatty acids in the rumen, while the proportions of propionic acid increased with a concomitant decrease in butyrate. Refaunation resulted in lower acetic acid and higher butyric acid proportions. The concentration of ammonia N in the rumen was clearly decreased after defaunation, already indicating an effect of the elimination of protozoa on nitrogen metabolism in the rumen. Defaunation also increased significantly the flow of total N, non ammonia N and individual and total amino acids in the proximal duodenum. Defaunation resulted in higher bacterial growth efficiency, significantly in one sheep, but the decrease after refaunation was statistically significant for both sheep. Determination of rumen digestibility of organic matter and acid detergent fibre (ADF) revealed lower values in the absence of the protozoa, while total digestibility was only influenced to a much lower extent. This indicated a shift of digestion from rumen to the lower digestive tract. Finally, earlier work is discussed in the light of the present findings.     

Effect of monensin on rumen ciliate protozoa in sheep

In a first experiment 4 rams consumed over a period of 70 days 0; 82; 208 and 345 mg of monensin daily, respectively. All doses of monensin caused a marked decrease in total numbers of rumen protozoa in samples taken after this period. More than 90% of total protozoal numbers belonged to the genus Entodinium. The sensitivity of the genus Entodinium to monensin was found to be lower than that of genera Dasytricha and Isotricha. In the second experiment 24 lambs were fed hay and concentrates in the ratio 60:40% (groups 1 and 2) and 40:60% (groups 3 and 4). Groups 2 and 4 received 40 mg monensin per animal daily during 16 weeks. The decrease in protozoal numbers due to monensin in samples taken after this period was significant (P less than 0.001) in lambs fed the concentrate diet (groups 3 and 4) and also in lambs from groups 1 and 2 fed the roughage diet (P less than 0.05). The statistical evaluation of the inhibition of total protozoal numbers by monensin (%) in lambs fed both diets has shown that the antiprotozoal effect of monensin was significantly more intensive with the concentrate diet (P less than 0.025) than with the roughage diet.     

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...     

Serum alpha-tocopherol concentration in sheep after intramuscular injection of DL-alpha-tocopherol.

Forty-three crossbred wethers weighing 35 to 60 kg were used to investigate the effect of a single i.m. injection of DL-alpha-tocopherol (DL-alpha-ol). Animals were offered 1 kg/d of a basal diet containing 25 ppm of vitamin E. Lambs were randomly assigned to one of five DL-alpha-ol injection treatments as follows: 1) control (placebo, 0 IU), 2) 125 IU, 3) 250 IU, 4) 500 IU, or 5) 1,000 IU. Blood samples were taken via jugular venipuncture on d 1 before treatment administration and thereafter at designated intervals up to 360 h postinjection. The i.m. injections of DL-alpha-ol irrespective of dose increased serum alpha-tocopherol. Results showed a dose x time interaction (P less than .0001) across all treatments. Serum alpha-tocopherol increased rapidly to maximum concentration during the first 8 to 12 h for all non-zero treatments, followed by a rapid decline to pretreatment values. The mean serum alpha-tocopherol concentration at 0 h was .69 microgram/mL. Estimated peak serum alpha-tocopherol concentrations +/- SE were 6.68 +/- 1.04, 9.62 +/- 1.04, 21.66 +/- 2.37, and 50.75 +/- 7.05 micrograms/mL for Treatments 2 through 5, respectively. Results showed a quadratic dose effect (P less than .0003) on maximum response with apparently no effect on time taken to reach this peak. There was also a quadratic dose effect (P less than .0001) on the area under the concentration-time curve. The time taken for serum alpha-tocopherol to return to pretreatment levels increased with dose (56, 64, 67, and 74 h, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Histomorphometric analysis of the rumen of sheep during development.

Histomorphometric and scanning electron microscopic analyses were carried out on 74 embryos and fetuses and 20 sheep (early postnatal to adult age). Histodifferentiation of the rumen took place at 33 days of fetal life. Ruminal pillars were observed at 42 days, and at 61 days, ruminal papillae appeared as evaginations of the epithelial stratum basale. Neutral mucopolysaccharides first appeared in epithelial cells at 46 days of fetal life; thereafter, numbers decreased gradually and subsequently stabilized in postnatal life. Acid mucopolysaccharides, mucins, and mucoid compounds were not detected. Age and diet were recognized as factors that determine the structure of the ruminal mucosa. Growth curves and formulas were set out for each tissue layer.     

Motility of the rumen after feeding sheep pelleted rations]

In an experiment with wethers the effect of the feeding with pelleted feed rations and the partial replacement of coarse fodder by non-treated beech sawdust on the motorial activity of the rumen was observed. The rumen motility was measured through a rumen fistula by means of the balloon method with the help of a capacitator primary unit, an electric manometer and a recording instrument. Over a period of 24 weeks the animals consumed 1.3 kg dry matter per day. It consisted of 41.8% meadow hay, 25.3% barley, 15.4% sawdust, 15.0% molasses, 1.3% urea, 0.76% mixed minerals and 0.48% hexametaphosphate in the form of pellets (test group) or the traditional classical form (control group). The feeding of pellets diminished the frequency (P less than 0.001) and the intensity of rumen contractions before and 1, 3 and 5 hours after feeding. Maximal frequency values were registered one hour after the food intake. During this time the number of secondary contractions of the rumen increased; differences of the frequency were, however, not registered, which means that the different physical form of the diet had no influence on the motorial activity of the rumen and that the food intake as such is the decisive factor. The diminished rumen motility in further hours after feeding was effected by treating the feed (grinding and pelleting).