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Ruminal synthesis, biohydrogenation, and digestibility of fatty acids by dairy cows.

Ruminal synthesis and biohydrogenation of fatty acids in dairy cows were determined by sampling duodenal digesta through T-cannulas. Fatty acid digestibility in the total tract also was measured. Five diets (concentrate:alfalfa hay:alfalfa haylage:corn silage, 2:1:1:1, DM) in a 5 x 5 Latin square contained either no added fat; 3 or 6% added calcium soap; or 3 or 6% animal-vegetable blend fat. Seventy percent of dietary fatty acids were recovered at the duodenum, and 106 g/d were synthesized in the rumen regardless of diets. Fatty acids synthesized in greatest amounts were odd or branched chains, whereas more than 90% of the fatty acids shorter than 14 carbons disappeared. Fatty acids in calcium soap were biohydrogenated 57% and in animal-vegetable blend 87%. Fatty acids in calcium soap were more digestible (80.0 vs. 75.7%) than those in the blended fat due to greater unsaturation in the small intestine. Ruminal microorganisms selectively synthesized fatty acids.

Animal Feed↗

Effect of nigericin, monensin, and tetronasin on biohydrogenation in continuous flow-through ruminal fermenters.

Four ionophores differing in cation selectivity were compared for their effect on microbial fermentation and biohydrogenation by ruminal bacteria in continuous culture. Monensin and nigericin are monovalent antiporters with selective binding affinities for Na+ and K+, respectively. Tetronasin is a divalent antiporter that binds preferentially with Ca2+ or Mg2+. Valinomycin is a monovalent uniporter and does not exchange K+ for H+. Steady-state concentrations of 2 micrograms/ml of monensin, nigericin, tetronasin, or valinomycin were maintained by constant infusion into fermenters. Molar percentages of acetate were lower, and those of propionate were higher, in the presence of monensin, nigericin, and tetronasin; all three ionophores also decreased CH4 production. Concentrations of valinomycin as high as 8 micrograms/ml had no effect on volatile fatty acids or CH4 production. Monensin, nigericin, and tetronasin inhibited the rate of biohydrogenation of linoleic acid. Continuous infusion of C18:2n-6 at a steady-state concentration of 314 micrograms/ml into fermenters receiving monensin, nigericin, or tetronasin resulted in lower amounts of stearic acid and higher amounts of oleic acid. Ionophores increased total C18:2 conjugated acids mainly because of an increase in the cis-9, trans-11-C18:2 isomer. If reflected in milk fat, ionophore-induced changes in ruminal lipids could enhance the nutritional qualities of milk.

Animals↗

Potential for biohydrogen and methane production from olive pulp.

The present study investigates the potential for thermophilic biohydrogen and methane production from olive pulp, which is the semi-solid residue coming from the two-phase processing of olives. It focussed on: a) production of methane from the raw olive pulp, b) anaerobic bio-production of hydrogen from the olive pulp, and c) subsequent anaerobic treatment of the hydrogen-effluent with the simultaneous production of methane. Both continuous and batch experiments were performed. The hydrogen potential of the olive pulp amounted to 1.6 mmole H2 per g TS. The methane potential of the raw olive pulp and hydrogen-effluent was as high as 19 mmole CH4 per g TS. This suggests that olive pulp is an ideal substrate for methane production and it shows that biohydrogen production can be very efficiently coupled with a subsequent step for methane production.

Anaerobiosis↗

Plasma fatty acids in sheep fed hydroxyethylsoyamide, a fatty acyl amide that resists biohydrogenation.

Hydroxyethylsoyamide (HESA) was made from soybean oil (SBO) and ethanolamine to determine the effectiveness of this fatty acyl amide to escape ruminal biohydrogenation and increase unsaturated fatty acids in plasma of sheep. The disappearance rate of 18:2n-6 from in vitro cultures was reduced 61% when the substrates contained added HESA compared to added linoleic acid. The decline in acetate to propionate ratio in the cultures also was less for HESA than for linoleic acid, which indicates lower inhibition of fermentation by HESA. Four sheep were housed in metabolism stalls for the collection of feces and urine and fed four diets in a Latin square design with 17-d periods. The diets contained no added fat (control) or were supplemented with 2.5% SBO, 5% butylsoyamide (BuSA), or 5% HESA. Plasma 18:2n-6 was not changed by feeding SBO. However, compared to the control diet, 18:2n-6 increased 19 and 35% in plasma fatty acids, and 74 and 113% in plasma triglycerides when sheep were fed BuSA and HESA, respectively. Digestibility of HESA was greater than for BuSA (99.5 vs. 48.4%, respectively). No amide was detected in plasma of sheep fed either BuSA or HESA. This study further supports that unsaturated oils can be converted to fatty acyl amides as a way to avoid ruminal biohydrogenation and elevate unsaturated fatty acids in body tissues of ruminants. Also, digestibility of the amide and plasma unsaturated fatty acids was enhanced more when the amide was synthesized from ethanolamine than when synthesized from butylamine.

Animals↗

Monitoring of microbial community structure and succession in the biohydrogen production reactor by denaturing gradient gel electrophoresis (DGGE).

To study the structure of microbial communities in the biological hydrogen production reactor and determine the ecological function of hydrogen producing bacteria, anaerobic sludge was obtained from the continuous stirred tank reactor (CSTR) in different periods of time, and the diversity and dynamics of microbial communities were investigated by denaturing gradient gel electrophoresis (DGGE). The results of DGGE demonstrated that an obvious shift of microbial population happened from the beginning of star-up to the 28th day, and the ethanol type fermentation was established. After 28 days the structure of microbial community became stable, and the climax community was formed. Comparative analysis of 16S rDNA sequences from reamplifying and sequencing the prominent bands indicated that the dominant population belonged to low G+C Gram-positive bacteria (Clostridium sp. and Ethanologenbacterium sp.), beta-proteobacteria (Acidovorax sp.), gamma-proteobacteria (Kluyvera sp.), Bacteroides (uncultured bacterium SJA-168), and Spirochaetes (uncultured eubacterium E1-K13), respectively. The hydrogen production rate increased obviously with the increase of Ethanologenbacterium sp., Clostridium sp. and uncultured Spirochaetes after 21 days, meanwhile the succession of ethanol type fermentation was formed. Throughout the succession the microbial diversity increased however it decreased after 21 days. Some types of Clostridium sp. Acidovorax sp., Kluyvera sp., and Bacteroides were dominant populations during all periods of time. These special populations were essential for the construction of climax community. Hydrogen production efficiency was dependent on both hydrogen producing bacteria and other populations. It implied that the co-metabolism of microbial community played a great role of biohydrogen production in the reactors.

Bacteroides↗

Molecular detection of the clostridia in an anaerobic biohydrogen fermentation system by hydrogenase mRNA-targeted reverse transcription-PCR.

Molecular biological approaches were developed to monitor the potential biohydrogen-producing clostridia in an anaerobic semisolid fermentation system that used brewery yeast waste as the fermentation substrate. The denaturing gradient gel electrophoresis with 16S rDNA gene-targeted polymerase chain reaction (PCR) analysis was employed to confirm the existence of clostridia in the system. Remarkably, reproducible nucleotide sequences of clostridia were obtained from different hydrogen production stages by using hydrogenase gene-targeted reverse transcription (RT)-PCR. These RNA-based information suggested that the predominant hydrogen-producing strains possess either a specific Clostridium pasteurianum-like or a specific Clostridium saccharobutylicum-like hydrogenase sequence. Comparison of the hydrogenase gene-targeted sequence profiles between PCR and RT-PCR revealed that the specific C. pasteurianum-like hydrogenase harboring bacterial strains were dominant in both mRNA and bacterial population level. On the other hand, the specific C. saccharobutylicum-like hydrogenase harboring strains expressed high level of hydrogenase mRNA but may not be dominant in population. Furthermore, quantitative real-time RT-PCR analysis showed the expression pattern of the clostridial hydrogenase mRNA and may serve as an activity index for the system.

Amino Acid Sequence↗

Metabolically engineered Rhodobacter sphaeroides RV strains for improved biohydrogen photoproduction combined with disposal of food wastes.

Three differently metabolically engineered strains, 2 single PHA- and Hup- mutants and one double PHA-/Hup- mutant, of the purple nonsulfur photosynthetic bacterium Rhodobacter sphaeroides RV, were constructed to improve a light-driven biohydrogen production process combined with the disposal of solid food wastes. These phenotypes were designed to abolish, singly or in combination, the competition of H2 photoproduction with polyhydroxyalkanoate (PHA) accumulation by inactivating PHA synthase activity, and with H2 recycling by abolishing the uptake hydrogenase enzyme. The performance of these mutants was compared with that of the wild-type strain in laboratory tests carried out in continuously fed photobioreactors using as substrates both synthetic media containing lactic acid and media from the acidogenic fermentation of actual fruit and vegetable wastes, containing mainly lactic acid, smaller amounts of acetic acia, and traces of higher volatile acids. With the lactic acid-based synthetic medium, the single Hup- and the double PHA-/Hup- mutants, but not the single PHA- mutant, exhibited increased rates of H2 photoproduction, about one third higher than that of the wild-type strain. With the food-waste-derived growth medium, only the single Hup- mutant showed higher rates of H2 production, but all 3 mutants sustained a longer-term H2 photoproduction phase than the wild-type strain, with the double mutant exhibiting overall the largest amount of H2 evolved. This work demonstrates the feasibility of single and multiple gene engineering of microorganisms to redirect their metabolism for improving H2 photoproduction using actual waste-derived substrates.

Acyltransferases↗

Partial inhibition of biohydrogenation of linoleic acid can increase the conjugated linoleic acid production of Butyrivibrio fibrisolvens A38.

Butyrivibrio fibrisolvens A38, one of the most active rumen bacteria in conjugated linoleic acid (CLA) production, was characterized in vitro. Previous findings that some inhibitory levels of substrate for biohydrogenation (BH) by B. fibrisolvens A38 resulted in more CLA accumulation led to a prediction that partial inhibition of BH could increase ruminal CLA production. The inhibitory conditions for bacterial growth were less effective on the isomerization step than on the following reduction step. Linoleic acid (LA) was inhibitory not only to cell growth but also to LA hydrogenation, and this effect was greater at high concentrations. The reduction step, converting CLA to hydrogenated products (trans-C18:1 and C18:0), was significantly inhibited, and more CLA accumulated during aerobic incubation when LA was added along with a glycolytic inhibitor, iodoacetate (IAA), to cells that were pre-adapted to LA (1 g/OD at 600 nm/L, P < 0.05). Monensin was more inhibitory than IAA to cell growth but less effective for CLA accumulation. Rumen fluid in the culture medium appeared to activate BH even in an aerobic condition, resulting in a lower CLA level than the control group (P < 0.05). Because the isomerization and reduction steps are coupled reactions in BH of most hydrogenating bacteria including B. fibrisolvens A38 cells, both positive and negative modulations of the reduction steps could be key determinants for CLA accumulation in the rumen.

Animals↗

The biohydrogenation of alpha-linolenic acid and oleic acid by rumen micro-organisms.

1. alpha-[U-(14)C]Linolenic acid was incubated with the rumen contents of sheep and the metabolic products were characterized by thin-layer chromatography, gas-liquid chromatography and absorption spectroscopy in the ultraviolet and infrared. 2. A tentative scheme for the biohydrogenation route to stearic acid is presented. The main pathway is through diconjugated cis-cis-cis-octadecatrienoic acid, non-conjugated trans-cis (cis-trans)-octadecadienoic acid and trans-octadecenoic acid, but other pathways are apparent. 3. Washed rumen micro-organisms possessed only a limited capacity to hydrogenate alpha-linolenic acid and oleic acid but the rate was greatly stimulated by a factor(s) present in the supernatant rumen liquor. 4. Pure cultures of Clostridium perfringens, Streptococcus faecalis, Escherichia coli and a coliform organism isolated from sheep faeces possessed negligible ability to hydrogenate unsaturated fatty acids compared with a mixed population of rumen micro-organisms. Butyrivibrio fibrisolvens slowly converted linoleic acid into octadecenoic acid.

Animals↗

Effects of treatment of whole fat soybeans or soy flour with formaldehyde to protect the polyunsaturated fatty acids from biohydrogenation in the rumen.

Full-fat, ground soy flour (GSF) was treated with 37% formaldehyde (HCHO) and evaluated by in vitro and in vivo criteria to determine the protection afforded linoleic acid against ruminal biohydrogenation when the materials described above were fed as a protein supplement to rations for growing lambs. The supplements compared were soybean meal (SBM), uked for 2 hours. Organoleptic evaluations were conducted to determine if any flavor differences in meat from lambs fed these supplements could be detected. Excellent protection of linoleic acid, the major polyunsaturated fatty acid in soybeans, was noted both in vitro and in vivo. Rump, shoulder, kidney knob and omental fat depots of lambs fed the HCHO treated GSF ration had significantly more linoleic acid than lambs fed untreated GSF while lambs fed untreated GSF had significantly, more linoleic acid in their fat depots than lambs fed SBM. Linoleic acid content of intramuscular (loin) fat from lambs fed HCHO treated GSF was not significantly different from lambs fed untreated GSF, but lambs fed untreated GSF had significantly more loin linoleic acid than lambs fed SBM. No significant differences were noted in daily feed intake, feed efficiency or average daily gain for lambs fed growing-finishing rations containing any of the products tested as the protein supplement. A taste panel could not detect any differences in flavor of ground loin among any of the treatments.

Adipose Tissue↗

Increased expression of a molecular chaperone GroEL in response to unsaturated fatty acids by the biohydrogenating ruminal bacterium, Butyrivibrio fibrisolvens.

Butyrivibrio fibrisolvens is the most active bacterial species in the biohydrogenation of polyunsaturated fatty acids (PUFA) in the rumen. It needs to remove the unsaturated bonds in order to detoxify the PUFA to enable the growth of the bacterium. Here, we investigated the response of cell membrane-associated proteins in B. fibrisolvens to growth in the presence of PUFA. Numerous changes were observed in the cell membrane-associated proteome. One of the main modifications occurring when the 18:2 fatty acids, linoleic acid and conjugated linoleic acid, were added, was an increased expression of the molecular chaperone GroEL.

Amino Acid Sequence↗

Biohydrogen production in continuous-flow reactor using mixed microbial culture.

The goal of the proposed project was to develop an anaerobic fermentation process that converts negative-value organic wastes into hydrogen-rich gas in a continuous-flow reactor under different operating conditions, such as hydraulic retention time (HRT), heat treatment, pH, and substrates. A series of batch tests were also conducted in parallel to the continuous study to evaluate the hydrogen conversion efficiency of two different organic substrates, namely sucrose and starch. A heat shock (at 90 degrees C for 15 minutes) was applied to the sludge in an external heating chamber known as a sludge activation chamber, as a method to impose a selection pressure to eliminate non-spore-forming, hydrogen-consuming bacteria and to activate spore germination. The experimental results showed that the heat activation of biomass enhanced hydrogen production by selecting for hydrogen-producing, spore-forming bacteria. The batch feeding at a shorter HRT of 20 hours (or higher organic loading rate) favored hydrogen production, whereas, at a longer HRT of 30 hours, methane was detected in the gas phase. The major organic acids of hydrogen fermentation were acetate, butyrate, and propionate. Up to 23.1% of influent chemical oxygen demand was consumed in biomass synthesis. Batch tests showed that the hydrogen-production potential of starch was lower than sucrose, and better conversion efficiency from starch was obtained at a lower pH of 4.5. However, addition of sucrose to starch improved the overall hydrogen-production potential and hydrogen-production rate. This study showed that sustainable biohydrogen production from carbohydrate-rich substrates is possible through heat activation of settled sludge.

Bacteria, Anaerobic↗

Influence of ruminal biohydrogenation on the feeding value of fat in finishing diets for feedlot cattle.

Four Holstein steers (212 kg) with cannulas in the rumen and proximal duodenum were used in a 4 x 4 Latin square experiment to study the influence of degree of ruminal biohydrogenation (BH) on the feeding value of supplemental fat. Treatments consisted of an 88% concentrate finishing diet supplemented with 1) 2% yellow grease (control); 2) 4% formaldehyde-protected fat (Rumentek), 2% yellow grease (LBH); 3) 2% Rumentek, 4% yellow grease (MBH); or 4) 6% yellow grease (HBH). Ruminal BH of HBH, MBH, and LBH diets was 74, 68, and 54%, respectively. High-fat supplementation decreased (7%, P < .05) intestinal digestibility of 18:0 but increased intestinal digestibility of 18:1 (3%, P < .10), 18:2 (14%, P < .01), and 18:3 (23%, P < .05). Increases in intestinal digestibility of 18:0 (quadratic effect, P < .05), 18:1 (linear effect, P < .01), 18:2 (linear effect, P < .01), 18:3 (linear effect, P < .05), and total fatty acids (linear effect, P < .05) were inversely related to BH. For every 1% increase in the proportion of 18:1 fat entering the small intestine, the digestibility of 18:0 increased 1%. High-fat supplementation depressed ruminal digestion of OM (11%, P < .05), NDF (16%, P < .05), starch (6%, P < .05), and feed N (12%, P < .01). Formaldehyde-protein protection of fat diminished its depressing effects on ruminal digestion of NDF (quadratic effect, P < .10) and enhanced ruminal escape of feed N (linear effect, P < .10). Postruminal digestion of OM was greater (4.6%, P < .10) for high-fat diets. High-fat diets decreased (P < .05) total tract digestion of OM (1.9%), NDF (7.4%), and starch (.5%). Postruminal and total tract digestibility of OM, NDF, N, and starch was not affected (P > .10) by BH. In a 125-d finishing trial, 100 yearling steers (362 kg) were used to evaluate treatment effects on growth performance. High-fat diets did not affect (P > .10) ADG but increased (P < .10) feed efficiency (9%, P < .10), dietary NEm (7.6%, P < .05), and dressing percentage (9%, P < .05). The magnitude of the increase in dressing percentage was inversely related (linear effect, P < .10) to BH. We conclude that decreasing ruminal BH will increase postruminal digestibility of fat, and hence the NE value of dietary fat. The synergistic effect of increasing the proportion of 18:1 on intestinal digestion of fat enables higher levels of fat supplementation. Protecting fat from BH minimizes the detrimental effects of supplemental fat on fiber digestion.

Animal Feed↗

Effects of forage and sunflower oil levels on ruminal biohydrogenation of fatty acids and conjugated linoleic acid formation in beef steers fed finishing diets.

Six Hereford steers (295 kg) cannulated in the proximal duodenum were used to evaluate the effects of forage and sunflower oil level on ruminal biohydrogenation (BH) and conjugated linoleic acid (CLA) outflow. Steers were fed one of six treatment diets in a 3 x 2 factorial arrangement of treatments (grass hay level: 12, 24, or 36% of DM; and sunflower oil level: 2 or 4% of DM) in a 6 x 6 Latin square design. The remainder of the diet was made up of steam rolled corn and protein/mineral supplement. Duodenal samples were collected for 4 d following 10-d diet adaptation periods. Data were analyzed with animal, period, forage level, sunflower oil level, and two-way interaction between forage and sunflower oil level in the model. Dry matter intake showed a quadratic response (P < 0.04), with an increase in DMI as forage level increased from 12 to 24% followed by a decrease in DMI when 36% forage was fed. Flow of fatty acids at the duodenum was higher (P < 0.03) for 4 vs. 2% sunflower oil diets, and similar among forage levels. Apparent ruminal digestibility of NDF increased in a linear manner (P < 0.04) as dietary forage level increased. Ruminal BH of dietary unsaturated 18-C fatty acids, oleic acid, and linoleic acid increased linearly (P < 0.05) as dietary forage level increased. Linoleic acid BH tended (P < 0.07) to be greater for 4 than 2% sunflower oil level. Duodenal flow of pentadecyclic, stearic, linolenic, and arachidic acids increased linearly (P < 0.05) as dietary forage level increased from 12 to 36%. Duodenal flow of linoleic acid decreased in a linear manner (P < 0.03) with increasing dietary forage level. Flow of trans-10 octadecenoate decreased linearly (P < 0.03) as dietary forage level increased, whereas trans-11 vaccenic acid flow to the duodenum increased (P < 0.01) linearly with increased dietary forage. Dietary forage or sunflower oil levels did not alter the outflow of cis-9, trans-11 CLA. Flows of cis-11, trans-13, and cis-9, cis-11 CLA increased linearly (P < 0.05) with increased dietary forage. Flows of cis-11, cis-13, and trans-11, trans-13 CLA decreased linearly (P < 0.05) with increased dietary forage. Increasing dietary forage levels from 12 to 36% in beef cattle finishing diets increased BH of unsaturated 18-C fatty acid and outflow of trans-11 vaccenic acid to duodenum without altering cis-9, trans-11 CLA outflow.

Animal Feed↗

Biohydrogenation of unsaturated fatty acids in continuous culture fermenters during digestion of orchardgrass or red clover with three levels of ground corn supplementation.

Diet digestibility and outputs of biohydrogenation intermediates were assessed in a continuous culture of ruminal microorganisms. Orchardgrass or red clover harvested and frozen during spring or fall served as the primary substrates for fermentation. During 10-d incubations, fermenters were fed thawed forage (50 g of DM/d), forage (42 g/d) plus 8 g/d of corn, or forage (34 g/d) plus 16 g/d of corn. Effluents from the last 3 d of incubation were composited for analyses. Starch input increased from 5 to 27% of DM as corn input increased from 0 to 16 g/d. Corn input reduced (P < 0.01) pH, increased (P < 0.01) microbial DM yield, and increased (P = 0.01) digestibility of DM, NDF, CP, and nonstructural carbohydrates. Overall, apparent hydrogenation (percentage) of cis9-18:1, 18:2n-6, and 18:3n-3 was greater (P < 0.05) with orchardgrass than clover. Hydrogenation of cis9-18:1 and 18:2n-6 increased (P = 0.01), but hydrogenation of 18:3n-3 decreased (P = 0.01) linearly due to corn input, regardless of forage. As a result, output of trans11, cis15-18:2 also decreased (P = 0.01). Average output of cis9,trans11-18:2 was greater (P = 0.01) for clover (1.3 mg/d) compared with orchardgrass (0.6 mg/d), but corn input with either forage increased (P = 0.01) cis9,trans11-18:2 output by 205%. Output of trans11-18:1 was greater (P = 0.01) from orchardgrass compared with clover (174 vs. 90 mg/d), but corn increased (P = 0.01) trans11-18:1 output only from clover fermentations. Output of trans10-18:1 was greater (P = 0.01) in response to orchardgrass compared with clover (10 vs. 4 mg/d), but corn addition doubled the output regardless of forage type. Output of trans10,cis12-18:2, which did not differ due to forage type, increased (P = 0.01) twofold in response to corn. Cis9,cis11-18:2 was a primary conjugated isomer produced from forage fermentations, but its output decreased (P = 0.03) in response to corn input. When inputs of 18:2n-6 plus 18:3n-3 were less than 0.9% of total DM (clover), hydrogenation was low (87%). When 18:2n-6 plus 18:3n-3 inputs were from 1.2 to 1.5% of total DM (orchardgrass), hydrogenation averaged 96%. Despite greater hydrogenation, incremental additions of cis9-18:1 and 18:2n-6 from corn grain increased (P < 0.05) outputs of trans10-18:1, trans11-18:1, trans10,cis12-18:2, cis9,trans11-18:2, and trans,trans-18:2 in effluent. Results suggest that forage species alone or in combination with corn grain can alter hydrogenation and profiles of intermediates to varying degrees.

Animal Feed↗

Effect of dietary vitamin E on rumen biohydrogenation pathways and milk fat depression in dairy cows fed high-fat diets.

Six lactating Holstein cows were assigned to a replicated Latin square design to test the effect of dietary vitamin E on milk fat depression and on the increased production of milk trans-10 C18:1 classically observed when feeding high doses of unsaturated fatty acids with low-fiber diets. Two diets (linseed diet and linseed diet + 12,000 IU of vitamin E/d) were compared during 2 periods of 21 d. The linseed diet presented a forage-to-concentrate ratio of 50:50 and contained extruded linseed (1.86 kg/d) and linseed oil (190 g/d). It was conceived to favor the "trans-11 to trans-10 shift" (low structural value and high level of unsaturated fatty acids). Milk yield and protein content were not affected by the diets. Milk of cows fed the linseed diet presented the typical symptoms of milk fat depression associated with a shift in biohydrogenation pathways: low fat content and high level of trans-10 C18:1. However, the high dose of dietary vitamin E provided significantly increased milk fat content (by 17.93%) and yield (by 15.56%) and decreased trans-10 C18:1 content (by 47.06%). In addition, it managed to significantly increase the daily yields of vaccenic (by 102.56%) and rumenic acids (by 56.67%). However, the sequence of administration of vitamin E influenced its effect, as vitamin E seemed to be more active in limiting the "trans-11 to trans-10 shift" when it was incorporated in the diet simultaneously with the fat. Once the shift had occurred, the subsequent addition of vitamin E was no longer able to completely counteract this process.

Animal Nutritional Physiological Phenomena↗

Biohydrogenation of unsaturated fatty acids. Hydrogenation by cell-free preparations of Butyrivibrio fibrisolvens.

Hydrogenation of cis-9,trans-11-octadecadienoic acid to yield trans-11-octadecenoic acid by cell-free preparations of Butyrivibrio fibrisolvens has been obtained under strictly anaerobic conditions. Reduced methyl viologen, NADH, and an endogenous electron donor each can serve as a reductant. Inhibition studies and gel filtration patterns reveal the presence of at least two hydrogenation systems, one of which is coupled through a flavin, possibly FMN. Although the enzymes comprising the biohydrogenation pathway, the fatty acid reductases and linoleic acid isomerase, are part of the bacterial membrane, they do not appear to be constituted as a multienzyme complex.

Cell-Free System↗

[Study on biohydrogen production by anaerobic biological fermentation of organic wasters].

Biological hydrogen production is a process of applying anaerobic biological fermentation method to produce hydrogen from organic wastes. Hydrogen production potential of different microbiological source and organic wastes, as well as the composition of fermentation product in liquid phase, were studied in the batch culture experiments. Magnifying experiment of biohydrogen production was carried out in 30 L improvement UASB reactor using anaerobic fermentation of artificial wastewater containing starch by activated sludge process. The experiment confirm that hydrogen and carbon dioxide percentage of biogas were 40%-51% and 49%-60% under the condition of hydrogen production process (pH was 4.5-5.5 sludge VSS was 10 g/L; starch-COD was 5000 mg/L), and no methane was observed. Hydrogen purity was more than 97% by an absorption of sodium hydrate solution, continuous hydrogen production was more than 120 days.

Bacteria, Anaerobic↗