Effect of oral nitroethane and 2-nitropropanol administration on methane-producing activity and volatile fatty acid production in the ovine rumen

Strategies are sought to reduce economic and environmental costs associated with ruminant methane emissions. The effect of oral nitroethane or 2-nitropropanol administration on ruminal methane-producing activity and volatile fatty acid production was evaluated in mature ewes. Daily administration of 24 and 72 mg nitroethane/kg body weight reduced (P<0.05) methane-producing activity by as much as 45% and 69% respectively, when compared to control animals given no nitroethane. A daily dose of 120 mg 2-nitropropanol/kg body weight was needed to reduce (P<0.05) methane-producing activity by 37% from that of untreated control animals. Reductions in methane-producing activity may have been diminished by the last day (day 5) of treatment, presumably due to ruminal adaptation. Oral administration of nitroethane or 2-nitropropanol had little or no effect on accumulations or molar proportions of volatile fatty acids in ruminal contents collected from the sheep. These results demonstrate that nitroethane was superior to 2-nitropropanol as a methane inhibitor and that both nitrocompounds reduced ruminal methanogenesis in vivo without redirecting the flow of reductant generated during fermentation to propionate and butyrate.     

Effect of select nitrocompounds on ruminal fermentation; an initial look at their potential to reduce economic and environmental costs associated with ruminal methanogenesis

Methane production by ruminal microbes during the digestion of feedstuffs is an inefficient process resulting in losses of 2-12% of the gross energy consumed by ruminants. Presently, we report the effect of three inhibitors on ruminal methane production in vitro. Mixed populations of ruminal microbes collected from cannulated cows maintained on an alfalfa hay:corn diet (50:50) were incubated at 39 degrees C for 24 h under a 100% carbon dioxide gas phase in closed tubes with 72 mM added sodium formate. Cultures were supplemented with 12 mM 2-nitropropanol, nitroethane or nitroethanol (experiment 1) or with 2, 12 or 24 mM nitroethane or a combination of 12 mM nitroethane and 4 mM nitroethanol (experiment 2). Control cultures containing no added nitrocompound were incubated simultaneously with treated incubations. Methane concentrations were reduced (P<0.05) from those measured in control incubations (27.6 +/- 2.1 and 17.7 +/- 0.8 micromol/ml; mean +/- SD for experiments 1 and 2, respectively) by at least 57% and as much as 94% in the nitrocompound supplemented incubations. By comparison, the widely fed methane inhibitor, monensin, typically reduces ruminal methane production by about 33%. Concentrations of volatile fatty acids and ammonia that accumulated in the nitrocompound supplemented incubations were not markedly affected compared to those produced by control cultures despite the reductions in methane produced. Hydrogen accumulated only slightly in cultures supplemented with the nitrocompounds. These results demonstrate that 2-nitropropanol, nitroethane and nitroethanol inhibit ruminal methane production. Further research is warranted to determine the mechanisms responsible for this inhibition and to see if these inhibitors can be used in practical application to reduce economic and environmental costs associated with ruminal methanogenesis.     

Effects of Dried Distillers’ Grain on Fecal Prevalence and Growth of Escherichia coli O157 in Batch Culture Fermentations from Cattle

Distillers’ grains (DG), a by-product of ethanol production, are fed to cattle. Associations between Escherichia coli O157 prevalence and feeding of DG were investigated in feedlot cattle (n = 379) given one of three diets: steam-flaked corn (SFC) and 15% corn silage with 0 or 25% dried distillers’ grains (DDG) or SFC with 5% corn silage and 25% DDG. Ten fecal samples were collected from each pen weekly for 12 weeks to isolate E. coli O157. Cattle fed 25% DDG with 5 or 15% silage had a higher (P = 0.01) prevalence of E. coli O157 than cattle fed a diet without DDG. Batch culture ruminal or fecal microbial fermentations were conducted to evaluate the effect of DDG on E. coli O157 growth. The first study utilized microbial inocula from steers fed SFC or dry-rolled corn with 0 or 25% DDG and included their diet as the substrate. Ruminal microbial fermentations from steers fed DDG had higher E. coli O157 contents than ruminal microbial fermentations from steers fed no DDG (P < 0.05) when no substrate was included. Fecal fermentations showed no DDG effect on E. coli O157 growth. In the second study with DDG as a substrate, ruminal fermentations with 0.5 g DDG had higher (P < 0.01) E. coli O157 concentrations at 24 h than ruminal fermentations with 0, 1, or 2 g DDG. In fecal fermentations, 2 g DDG resulted in a higher concentration (P < 0.05) at 24 h than 0, 0.5, or 1 g DDG. The results indicate that there is a positive association between DDG and E. coli O157 in cattle, and the findings should have important ramifications for food safety.     

Effect of protein supplementation on ruminal parameters and microbial community fingerprint of Nellore steers fed tropical forages

In tropical regions, protein supplementation is a common practice in dairy and beef farming. However, the effect of highly degradable protein in ruminal fermentation and microbial community composition has not yet been investigated in a systematic manner. In this work, we aimed to investigate the impact of casein supplementation on volatile fatty acids (VFA) production, specific activity of deamination (SAD), ammonia concentration and bacterial and archaeal community composition. The experimental design was a 4×4 Latin square balanced for residual effects, with four animals (average initial weight of 280±10 kg) and four experimental periods, each with duration of 29 days. The diet comprised Tifton 85 (Cynodon sp.) hay with an average CP content of 9.8%, on a dry matter basis. Animals received basal forage (control) or infusions of pure casein (230 g) administered direct into the rumen, abomasum or divided (50 : 50 ratio) in the rumen/abomasum. There was no differences (P>0.05) in ruminal pH and microbial protein concentration between supplemented v. non-supplemented animals. However, in steers receiving ruminal infusion of casein the SAD and ruminal ammonia concentration increased 33% and 76%, respectively, compared with the control. The total concentration of VFA increased (P<0.05) in steers receiving rumen infusion of casein. SAD and the microbial protein concentration did not vary significantly among treatments during the feeding cycle, but mean SAD values were greater in steers supplemented in the rumen and rumen/abomasum. Ruminal ammonia concentration was positively correlated with SAD in animals receiving ruminal infusion of casein. Polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE) analysis revealed low similarity between treatments, animals and time of sample collection. Richness analysis and determination of the Shannon–Wiener index indicated no differences (P>0.05) in species richness and diversity of γ-proteobacteria, firmicutes and archaea between non-supplemented Nellore steers and steers receiving casein supplementation in the rumen. However, species richness and the Shannon–Wiener index were lower (P<0.05) for the phylum bacteroidetes in steers supplemented with casein in the rumen compared with non-supplemented animals. Venn diagrams indicated that the number of unique bands varied considerably among individual animals and was usually higher in number for non-supplemented steers compared with supplemented animals. These results add new knowledge about the effects of ruminal and postruminal protein supplementation on metabolic activities of rumen microbes and the composition of bacterial and archaeal communities in the rumen of steers.     

Effects of dried distillers' grain on fecal prevalence and growth of Escherichia coli O157 in batch culture fermentations from cattle

Distillers' grains (DG), a by-product of ethanol production, are fed to cattle. Associations between Escherichia coli O157 prevalence and feeding of DG were investigated in feedlot cattle (n = 379) given one of three diets: steam-flaked corn (SFC) and 15% corn silage with 0 or 25% dried distillers' grains (DDG) or SFC with 5% corn silage and 25% DDG. Ten fecal samples were collected from each pen weekly for 12 weeks to isolate E. coli O157. Cattle fed 25% DDG with 5 or 15% silage had a higher (P = 0.01) prevalence of E. coli O157 than cattle fed a diet without DDG. Batch culture ruminal or fecal microbial fermentations were conducted to evaluate the effect of DDG on E. coli O157 growth. The first study utilized microbial inocula from steers fed SFC or dry-rolled corn with 0 or 25% DDG and included their diet as the substrate. Ruminal microbial fermentations from steers fed DDG had higher E. coli O157 contents than ruminal microbial fermentations from steers fed no DDG (P < 0.05) when no substrate was included. Fecal fermentations showed no DDG effect on E. coli O157 growth. In the second study with DDG as a substrate, ruminal fermentations with 0.5 g DDG had higher (P < 0.01) E. coli O157 concentrations at 24 h than ruminal fermentations with 0, 1, or 2 g DDG. In fecal fermentations, 2 g DDG resulted in a higher concentration (P < 0.05) at 24 h than 0, 0.5, or 1 g DDG. The results indicate that there is a positive association between DDG and E. coli O157 in cattle, and the findings should have important ramifications for food safety.     

Trace mineral source influences digestion,ruminal fermentation,and ruminal copper,zinc, and manganese distribution in steers fed a diet suitable for lactating dairy cows

High solubility of certain trace minerals (TM) in the rumen can alter nutrient digestibility and fermentation. The objectives of the present studies were to determine the effects of TM source on 1) nutrient digestibility and ruminal fermentation, 2) concentrations of soluble Cu, Zn, and Mn in the rumen following a pulse dose of TM, and 3) Cu, Zn, and Mn binding strength on ruminal digesta using dialysis against a chelating agent in steers fed a diet formulated to meet the requirements of a high producing dairy cow. Twelve Angus steers fitted with ruminal cannulae were adapted to a diet balanced with nutrient concentrations similar to a diet for a high producing lactating dairy cow for 21 d. Steers were then randomly assigned to dietary treatments consisting of 10 mg Cu, 40 mg Mn, and 60 mg Zn/kg DM from either sulfate (STM), hydroxychloride (HTM) or complexed trace minerals (CTM). The experimental design did not include a negative control (no supplemental Cu, Mn, or Zn) because the basal diet did not meet the National Research Council requirement for Cu and Zn. Copper, Mn, and Zn are also generally supplemented to lactating dairy cow diets at concentrations approximating those supplied in the present study. Following a 14-d adaptation period, total fecal output was collected for 5-d. Following the fecal collection period, rumen fluid was collected for Volatile fatty acid (VFA) parameters. On the following day, the same diet was provided for 14 d, without supplemental Cu, Zn, and Mn. This period served as a wash-out period. A pulse dose of 100, 400, and 600 mg of Cu, Zn, Mn, respectively, from either STM, HTM, or CTM, was administered via ruminal cannulae to the steers on day 15. Over a 24-h period ruminal samples were obtained every 2-h. Following centrifugation, the supernatant was analyzed for Cu, Mn, and Zn. Ruminal solid digesta samples from times 0, 12, and 24 h after bolus dosing were exposed to dialysis against Tris-EDTA. Digestibility of NDF and ADF were lesser in STM vs. HTM and vs. CTM supplemented steers. Steers receiving HTM and CTM had greater total VFA concentrations than STM, and molar proportions of individual VFA were not affected by treatment. Ruminal soluble Cu and Zn concentrations were greater post dosing in STM and CTM supplemented steers at 2, 4, and 6 h for Cu and 4, 6, 8, 10 and 12 h for Zn when compared to HTM supplemented steers. The release of Cu and Zn from ruminal solid digesta following dialysis against Tris-EDTA at 12 and 24 h postdosing was greater for steers receiving HTM compared to those receiving STM or CTM. Results indicate trace mineral source impacts: 1) how tightly bound Cu and Zn are to ruminal solid digesta; 2) fiber digestion; 3) and ruminal total VFA concentrations.     

Effects of feed withdrawal duration on animal behaviour,rumen microbiota and blood chemistry in feedlot cattle: implications for rumen acidosis

Feed withdrawal (FW) is a frequent issue in open outdoor feedlot systems, where unexpected circumstances can limit the animals’ access to food. The relationship among fasting period, animal behaviour during feed reintroduction (FR) and acidosis occurrence has not been completely elucidated. Twenty steers fitted with rumen catheters were fed a high-concentrate diet (concentrate : forage ratio 85 : 15) and were challenged by a protocol of FW followed by FR. The animals were randomly assigned to one of the four treatments: FW for 12 h (T12), 24 h (T24), 36 h (T36) or no FW (control group) followed by FR. The steers’ behaviour, ruminal chemistry, structure of the ruminal microbial community, blood enzymes and metabolites and ruminal acidosis status were assessed. Animal behaviour was affected by the FW–FR challenge ( P < 0.05). Steers from the T12, T24 and T36 treatments showed a higher ingestion rate and a lower frequency of rumination. Although all animals were suspected to have sub-acute ruminal acidosis (SARA) prior to treatment, a severe case of transient SARA arose after FR in the T12, T24 and T36 groups. The ruminal pH remained below the threshold adopted for SARA diagnosis ( pH value = 5.6) for more than three consecutive hours (24, 7 and 19 h in the T12, T24 and T36 treatments, respectively). The FW–FR challenge did not induce clinical acute ruminal acidosis even though steers from the T36 treatment presented ruminal pH values that were consistent with this metabolic disorder (pH threshold for acute acidosis = 5.2). Total mixed ration reintroduction after the withdrawal period reactivated ruminal fermentation as reflected by changes in the fermentation end-products. Ruminal lactic acid accumulation in steers from the T24 and T36 treatments probably led to the reduction of pH in these groups. Both the FW and the FR phases may have altered the structure of the ruminal microbiota community. Whereas fibrolytic bacterial groups decreased relative abundance in the restricted animals, both lactic acid producer and utiliser bacterial groups increased ( P < 0.05). The results demonstrated a synchronisation between Streptococcus (lactate producer) and Megasphaera (lactate utiliser), as the relative abundance of both groups increased, suggesting that bacterial resilience may be central for preventing the onset of metabolic disturbances such as ruminal acidosis. A long-FW period (36 h) produced rumen pH reductions well below and lactic acid concentration increased well above the accepted thresholds for acute acidosis without any perceptible clinical signs.     

Effects of nitro compounds and feedstuffs on in vitro methane production in chicken cecal contents and rumen fluid

Short-chain volatile fatty acids (VFA) and methane are the products from a wide variety of microorganisms living in the gastrointestinal tract. The objective of this study was to examine effects of feedstuff and select nitro compounds on VFA and methane production during in vitro incubation of laying hen cecal contents and rumen fluid from cattle and sheep. In the first experiment, one of the three nitro compound was added to incubations containing cecal contents from laying hens supplemented with either alfalfa (AF) or layer feed (LF). Both feed material influenced VFA production and acetic acid was the primary component. Incubations with nitro ethanol and 2-nitropropanol (NP) had significantly (P<0.05) higher propionate concentrations than incubations with added nitroethane (NE). The results further indicated that incubations containing LF produced significantly (P<0.05) more butyrate than incubations with added AF. Addition of NP and LF to incubations of avian cecal flora may promote Gram-positive, saccharolytic, VFA-producing bacteria, especially Clostridium spp. which is the predominant group in ceca. Similar to VFA production, both feed materials fostered methane production in the incubations although methane was lower (P<0.05) in incubations with added nitro compound, particularly NE. In experiments 3-8, NE was examined in incubations of bovine or ovine rumen fluid or cecal contents containing either AF or LF. Unlike cecal contents, LF significantly (P<0.05) supported in vitro methane production in incubations of both rumen fluids. The results show that NE impedes methane production, especially in incubations of chicken cecal contents.     

Moderation of ruminal fermentation by ciliated protozoa in cattle fed a high-grain diet.

The objective of this study was to assess the influence of ciliated protozoa on ruminal fermentation in cattle fed high-grain diets. Six ruminally cannulated steers fed a corn-based grain diet (85% concentrate plus 15% alfalfa hay) at 12-h intervals were assigned randomly to two groups, ciliate free and faunated, in a crossover design. Defaunation was by ruminal emptying, omasal flushing, and treatment with sodium sulfosuccinate. Two to 3 weeks after defaunation, the ruminal contents of all steers were sampled before the morning feeding (0 h) and at 1, 2, 4, 6, 8, and 12 h after feeding to measure pH, analyze fermentation products, and monitor counts of ciliated protozoa and lactic acid-producing and -fermenting bacterial groups. Total numbers of ciliated protozoa in the faunated steers averaged 4.3 x 10(5)/g, and the protozoa consisted of nine genera. Ciliate-free steers had lower (P less than 0.01) ruminal pHs (pH 5.97) than faunated cattle (pH 6.45); however, the treatment-time interaction was not significant. Ruminal lactate and ammonia concentrations were similar in both groups. The total volatile fatty acid concentration was higher (P less than 0.05) in the ciliate-free steers than in the faunated steers and exhibited a treatment-time interaction (P less than 0.05). The acetate-to-propionate ratio was higher (P less than 0.05) in the faunated group than in the ciliate-free group and showed a treatment-time interaction (P less than 0.05). Total anaerobic bacterial counts were about fourfold higher in the ciliate-free group than in the faunated group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Moderation of ruminal fermentation by ciliated protozoa in cattle fed a high-grain diet.

The objective of this study was to assess the influence of ciliated protozoa on ruminal fermentation in cattle fed high-grain diets. Six ruminally cannulated steers fed a corn-based grain diet (85% concentrate plus 15% alfalfa hay) at 12-h intervals were assigned randomly to two groups, ciliate free and faunated, in a crossover design. Defaunation was by ruminal emptying, omasal flushing, and treatment with sodium sulfosuccinate. Two to 3 weeks after defaunation, the ruminal contents of all steers were sampled before the morning feeding (0 h) and at 1, 2, 4, 6, 8, and 12 h after feeding to measure pH, analyze fermentation products, and monitor counts of ciliated protozoa and lactic acid-producing and -fermenting bacterial groups. Total numbers of ciliated protozoa in the faunated steers averaged 4.3 x 10(5)/g, and the protozoa consisted of nine genera. Ciliate-free steers had lower (P less than 0.01) ruminal pHs (pH 5.97) than faunated cattle (pH 6.45); however, the treatment-time interaction was not significant. Ruminal lactate and ammonia concentrations were similar in both groups. The total volatile fatty acid concentration was higher (P less than 0.05) in the ciliate-free steers than in the faunated steers and exhibited a treatment-time interaction (P less than 0.05). The acetate-to-propionate ratio was higher (P less than 0.05) in the faunated group than in the ciliate-free group and showed a treatment-time interaction (P less than 0.05). Total anaerobic bacterial counts were about fourfold higher in the ciliate-free group than in the faunated group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pathway of oxidation of pyruvic oxime by a heterotrophic nitrifier of the genus Alcaligenes: evidence against nitroethane as an intermediate

The role of nitroethane as an intermediate in the oxidation of pyruvic oxime to nitrate by an Alcaligenes sp. was examined. Unlike pyruvic oxime, which serves as a sole source of C and N for the bacterium, nitroethane was incapable of supporting the growth of the microbe. Nitroethane was metabolized and diauxic growth did occur, however, if the nitroethane medium was amended with yeast extract. Alcaligenes sp. resting cells and cell-free extracts were prepared from nitroethane-yeast extract grown cultures and the maximum rate of nitrite synthesis when nitroethane was the substrate was 6.8 nmol min-1 mg cell protein-1, a 10-fold lower rate than that previously noted for pyruvic oxime oxidation. These cell-free extracts were unable to metabolize pyruvic oxime. Resting cells and cell-free extracts prepared from Alcaligenes sp. cells grown in a pyruvic oxime medium were, conversely, incapable of metabolizing nitroethane. Collectively, these results indicate that nitroethane is not an intermediate in the pathway of pyruvic oxime oxidation and that two separate enzyme systems exist in the Alcaligenes sp. for the metabolism of pyruvic oxime and nitroethane.     

Microbial assimilation of alkyl nitro compounds and formation of nitrite

66 representative strains of bacteria, yeasts and fungi were tested for their ability to grow in a semidefined medium containing 0.5% nitroethane as a nitrogen source. About half of them were found capable of growing in the medium. Hansenula beijerinckii, Candida utilis, and Penicillium chrysogenum were most active in assimilating nitroethane. 2-Nitropropane inhibited growth of most of the microorganisms tested in a medium containing 0.2% peptone and 0.2% glycerol. Hansenula mrakii was found to grow rapidly in the nitroethane-peptone medium after a lag phase. Nitrite was accumulated in the culture fluid after the phase of logarithmic multiplication, and increased with increase of the growth, followed by a decline after the maximum growth. The alkyl nitro compounds were oxidatively denitrified to form nitrite by the crude enzyme from Hansenula mrakii. Nitroethane was generally a poor substrate, but was the best inducer to produce the nitro compounds oxidizing enzyme. 2-Nitropropane and nitroethane were enzymatically oxidized to and acetone and acetaldehyde, respectively, which were isolated as 2,4-dinitrophenylhydrazones and identified. Nitrite formed was found to be reduced into ammonia by the intact cells and also the crude enzyme.     

Impact of adding nitrate or increasing the lipid content of two contrasting diets on blood methaemoglobin and performance of two breeds of finishing beef steers

Adding nitrate to the diet or increasing the concentration of dietary lipid are effective strategies for reducing enteric methane emissions. This study investigated their effect on health and performance of finishing beef cattle. The experiment was a two×two×three factorial design comprising two breeds (CHX, crossbred Charolais; LU, Luing); two basal diets consisting of (g/kg dry matter (DM), forage to concentrate ratios) 520 : 480 (Mixed) or 84 : 916 (Concentrate); and three treatments: (i) control with rapeseed meal as the main protein source replaced with either (ii) calcium nitrate (18 g nitrate/kg diet DM) or (iii) rapeseed cake (RSC, increasing acid hydrolysed ether extract from 25 to 48 g/kg diet DM). Steers (n=84) were allocated to each of the six basal diet×treatments in equal numbers of each breed with feed offered ad libitum. Blood methaemoglobin (MetHb) concentrations (marker for nitrate poisoning) were monitored throughout the study in steers receiving nitrate. After dietary adaptation over 28 days, individual animal intake, performance and feed efficiency were recorded for a test period of 56 days. Blood MetHb concentrations were low and similar up to 14 g nitrate/kg diet DM but increased when nitrate increased to 18 g nitrate/kg diet DM (P<0.001). An interaction between basal diet and day (P<0.001) indicated that MetHb% was consistently greater in Concentrate – than Mixed-fed steers at 18 g nitrate/kg diet DM. Maximum individual MetHb% was 15.4% (of total Hb), which is lower than considered clinically significant (30%). MetHb concentrations for individual steers remained consistent across time. Concentrate-fed steers were more efficient (lower residual feed intake (RFI) values) than Mixed-fed steers (P<0.01), with lower dry matter intake (DMI) (kg/day) (P<0.001) and similar average daily gain (ADG). CHX steers were more efficient (lower RFI; P<0.01) than LU steers with greater ADG (P<0.01), lower DMI (/kg BW; P<0.01) and lower fat depth (P<0.001). ADG, BW or DMI did not differ across dietary treatments (P>0.05). Neither basal diet nor treatment affected carcass quality (P>0.05), but CHX steers achieved a greater killing out proportion (P<0.001) than LU steers. Thus, adding nitrate to the diet or increasing the level of dietary lipid through the use of cold-pressed RSC, did not adversely affect health or performance of finishing beef steers when used within the diets studied.     

溶菌酶对瘤胃体外发酵、甲烷生成及微生物菌群结构的影响

【目的】通过体外静态模拟瘤胃发酵法研究溶菌酶对瘤胃发酵、甲烷生成及微生物菌群结构的影响。【方法】采用单因素多水平试验设计,溶菌酶添加水平分别为0(L-0,对照组)、0.1 mg/100 m L(L-0.1)、1 mg/100 m L(L-1)、10 mg/100 m L(L-10)和100 mg/100 m L(L-100),定时测定产气量和甲烷产量,培养24 h后,发酵液用于发酵参数和微生物菌群数量的q PCR测定,其中L-0、L-1和L-100三个组发酵液同时进行16S r RNA基因Illumina高通量测序。【结果】与对照组相比,低剂量溶菌酶添加(L-0.1组)不影响甲烷产量、氨氮浓度、干物质消失率、有机物消失率和总挥发性脂肪酸等瘤胃发酵参数(P0.05);随着剂量提高,L-1处理组甲烷产量、氨氮浓度显著降低(P0.05),丙酸浓度显著增加(P0.05),并且干物质消失率、有机物消失率和总挥发性脂肪酸不受影响(P0.05);而较高剂量组(L-10和L-100组)虽然甲烷产量显著降低,丙酸浓度显著增加(P0.05),但干物质消失率和有机物消失率也显著降低(P0.05)。q PCR结果显示高剂量组(L-100组)总菌、原虫、甲烷菌数量与对照组相比显著降低(P0.05),而L-0.1、L-1和L-10组总菌、真菌和原虫数量与对照组相比均无显著变化(P0.05)。高通量测序主成分分析(PCA)显示对照组与溶菌酶添加组间瘤胃细菌组成的明显区分,说明添加溶菌酶显著改变了瘤胃细菌菌群结构。溶菌酶通过增加月形单胞菌和琥珀酸弧菌等丙酸生成菌的相对丰度,使更多的氢被用于生成丙酸,导致甲烷产量降低;溶菌酶可抑制普雷沃氏菌和拟杆菌属等蛋白降解菌的生长,进而减少蛋白质过度降解,降低氨氮浓度。【结论】添加适宜浓度(1 mg/100 m L)的溶菌酶可通过调控瘤胃微生态改变瘤胃发酵模式,降低瘤胃甲烷和氨的生成,短期内并不影响饲料消化。     

儿童急性淋巴细胞白血病对预后的影响分析

目的:探讨儿童急性淋巴细胞白血病分型对预后的影响,为临床治疗提供依据.方法:回顾性分析2007年1月～2008年12月我院收治的急性淋巴细胞白血病患儿32例,比较不同分型的预后情况.结果:ALL-L1、ALL-L2与ALL-L3的首次诱导CR率(X2=1.087,P＞0.05)、完全CR率(X2=0.607,P＞0.05),差异无统计学意义;CR的治疗时间(t=6.001,P＜0.05)、3年生存率(X2=9.458,P＜0.05),差异有统计学意义.T-ALL、B-ALL的首次诱导CR率(X2=8.891,P＜0.05)、达到CR治疗时间(t=6.361,P＜0.05)、完全CR率(X2=11.892,P＜0.05),差异有统计学意义.两型的3年生存率(X2=1.536,P＞0.05),差异有统计学意义.B-ALL中各型别首次诱导CR率(X2=0.494,P＞0.05)和完全CR率(X2=0.405,P＞0.05),差异统计学意义.B-ALL中各型别达到CR的治疗时间(t=7.007,P＜0.05)和3年生存率(X2=6.609,P＜0.05),差异有统计学意义.结论:儿童急性白血病其预后与其分型有一定的相关性,因此临床治疗应结合患儿的分型进行个体化治疗.     

Effects of Exogenous Melatonin and Tryptophan on Fecal Shedding of <Emphasis Type="Italic">E. Coli</Emphasis> O157:H7 in Cattle

Fecal prevalence of Escherichia coli O157 in ruminants is highest in the summer decreasing to very low levels in the winter. We hypothesize that this seasonal variation is a result of physiological responses within the host animal to changing day-length. To determine the effects of melatonin (MEL) on fecal shedding of E. coli O157:H7 in cattle, eight crossbred beef steers identified as shedding E. coli O157:H7, were allotted to treatment: control or MEL (0.5 mg/kg body weight (BW); 1×) administered orally daily for 7 days. After a 5-day period of no treatment, a second MEL dose (5.0 mg/kg BW; 10×) was administered daily for 4 days. Fecal samples were collected daily for qualification of E. coli O157:H7. No differences (P > 0.10) were observed in the percentage of E. coli O157:H7 positive fecal samples in steers receiving the 1× MEL dose, however the 10× dose decreased (P = 0.05) the percentage of fecal samples E. coli O157:H7 positive. Serum MEL concentrations were higher in the 1×, but not 10×, treated animals compared to control animals. Although it is difficult to explain, this may be a result of decreasing day-length increasing serum melatonin concentrations that may have masked any treatment effect on serum melatonin. In a second similar experiment, a second group of cattle (heifers and steers) were administered tryptophan (TRP) over a 17-day experimental period (5 g/head/day for 10 days followed by 10 g/head/day for 7 days). Tryptophan had no effect (P > 0.20) on the percentage of fecal samples positive for E. coli O157. Serum TRP (P < 0.05), but not MEL (P > 0.20), concentrations were elevated in TRP-treated animals. The decrease in the number of positive fecal samples observed in the first experiment, may be related to gastrointestinal MEL, affected by the 10×, but not 1× MEL dose.     

Effect of different levels of concentrate on ruminal microorganisms and rumen fermentation in Nellore steers

The aim of this study was to investigate the effect of different dietary levels of concentrate on feed intake, digestibility, ruminal fermentation and microbial population in steers. Eight Nellore steers fitted with ruminal cannulas were used in a double 4 × 4 Latin square design experiment. The dietary treatments consist of four different proportions of concentrate to roughage: 30:70, 40:60, 60:40 and 80:20% in the dry matter, resulting in Diets 30, 40, 60 and 80, respectively. The roughage was corn silage, and the concentrate was composed of corn, soybean meal and urea. Apparent digestibility of organic matter and crude protein showed a linear association with concentrate proportion (p = 0.01), but the increased concentrate levels did not affect the digestibility of fibre. The lowest ruminal pH-values were observed in animals fed with Diet 80, remaining below pH 6.0 from 6 h after feeding, while in the other diets, the ruminal pH was below 6.0 not before 12 h after feeding. After feeding Diet 80, the ammonia concentration in the rumen was significantly the highest. Higher dietary concentrate levels resulted in a linear increase of propionic acid concentrations, a linear reduction of the ratio acetic acid to propionic acid (p < 0.01) and a linear increased synthesis of microbial nitrogen (p < 0.001). The predicted production of methane was lower in diets with greater amounts of concentrate (p = 0.032). The population of methanogens, R. flavefaciens and R. albus decreased with higher concentrate levels, while the population of S. ruminantium increased (p < 0.05). The results indicate that greater amounts of concentrate do not decrease ruminal pH-values as much as expected and inhibit some cellulolytic bacteria without impairing the dry matter intake and fibre digestibility in Nellore steers.     

The impact of divergent breed types and diets on methane emissions,rumen characteristics and performance of finishing beef cattle

This study was undertaken to further develop our understanding of the links between breed, diet and the rumen microbial community and determine their effect on production characteristics and methane (CH₄) emissions from beef cattle. The experiment was of a 2×2 factorial design, comprising two breeds (crossbred Charolais (CHX); purebred Luing (LU)) and two diets (concentrate-straw or silage-based). In total, 80 steers were used and balanced for sire within each breed, farm of origin and BW across diets. The diets (fed as total mixed rations) consisted of (g/kg dry matter (DM)) forage to concentrate ratios of either 500 : 500 (Mixed) or 79 : 921 (Concentrate). Steers were adapted to the diets over a 4-week period and performance and feed efficiency were then measured over a 56-day test period. Directly after the 56-day test, CH₄ and carbon dioxide (CO₂) emissions were measured (six steers/week) over a 13-week period. Compared with LU steers, CHX steers had greater average daily gain (ADG; P<0.05) and significantly (P<0.001) lower residual feed intake. Crossbred Charolais steers had superior conformation and fatness scores (P<0.001) than LU steers. Although steers consumed, on a DM basis, more Concentrate than Mixed diet (P<0.01), there were no differences between diets in either ADG or feed efficiency during the 56-day test. At slaughter, however, Concentrate-fed steers were heavier (P<0.05) and had greater carcass weights than Mixed-fed steers (P<0.001). Breed of steer did not influence CH₄ production, but it was substantially lower when the Concentrate rather than Mixed diet was fed (P<0.001). Rumen fluid from Concentrate-fed steers contained greater proportions of propionic acid (P<0.001) and lower proportions of acetic acid (P<0.001), fewer archaea (P<0.01) and protozoa (P=0.09), but more Clostridium Cluster XIVa (P<0.01) and Bacteroides plus Prevotella (P<0.001) than Mixed-fed steers. When the CH₄ to CO₂ molar ratio was considered as a proxy method for CH₄ production (g/kg DM intake), only weak relationships were found within diets. In conclusion, although feeding Concentrate and Mixed diets produced substantial differences in CH₄ emissions and rumen characteristics, differences in performance were influenced more markedly by breed.     

Heterotrophic nitrification by Pseudomonas aeruginosa

Summary Several soil bacteria and fungi produce nitrite when provided with acetaldoxime. Nitrite formation by one isolate, identified as a strain of Pseudomonas aeruginosa, is not directly linked to growth but rather proceeds mainly after the active growth period. The added oxime-nitrogen is converted completely to nitrite, and nitrate is not formed. Extracts of the bacterium generate nitrite, but not nitrate, more rapidly from nitroethane than from the added oxime. The enzyme system catalyzing the formation of nitrite in oxime solutions is soluble and inducible, whereas the enzyme catalyzing the release of equimolar quantities of nitrite and acetaldehyde from nitroethane is constitutive. The slow rate of nitrite production when the enzyme preparation is provided with acetaldoxime is not markedly increased by added cofactors. The soluble enzymes also generate nitrite when incubated with several aliphatic and alicyclic oximes and nitro compounds. Nitroethane is not formed from acetaldoxime. The possible mechanism of this nitrification reaction catalyzed by a heterotrophic microorganism is discussed.     

Effect of soybean oil availabilities on rumen biohydrogenation and duodenal flow of fatty acids in beef cattle fed a diet with crude glycerine

Soybean oil with different ruminal availability (whole soybeans (WS), soybean oil (SO) and calcium salts (CS)) was used to evaluate the fatty acid (FA) intake, rumen biohydrogenation (BH) and duodenal flow of FA in Nellore steers fed diets with crude glycerine (CG). Eight castrated Nellore steers were fitted with a ruminal and duodenal silicone cannula, and distributed in a double, simultaneous, Latin square 4 × 4 design with four diets and four experimental periods. Concentrates contained ground maize, urea, mineral salts, CG (100 g/kg DM) and soybean products with different availability of soybean oil: (1) no additional fat (CO), (2) WS, (3) SO or (4) CS. Fat supplementation was fixed to obtain 50 g ether extract/kg DM. Experimental treatments had no effect on DM intake, DM duodenal flow or ruminal turnover rate of C:16 FA. However, fat addition increased C:18 and turnover rates of total FA rumen (p < 0.05). CS resulted in lower C:18 turnover rates and lower ruminal BH of monounsaturated and unsaturated FA (UFA) than WS (p < 0.05). SO resulted in a greater duodenal flow of C18:0 (stearic acid), C18:1t-11 (vaccenic acid) and saturated FA than the WS and CS diets (p < 0.05). CS resulted in a higher duodenal flow of C18:3n-3 (linolenic acid) than WS (p < 0.05). The association of CG and calcium salts in Nellore steers was the best nutritional strategy to increase duodenal flow of healthier UFA, which may increase the deposition of these FA in meat. However, SO associated with CG association increased the duodenal flow of vaccenic acid, which is main precursor of endogenous synthesis of conjugated linoleic acids in tissues.