Rhodobacter sphaeroides mutants which accumulate 5-aminolevulinic acid under aerobic and dark conditions

The photosynthetic bacterium Rhodobacter sphaeroides accumulates 5-aminolevulinic acid (ALA), which is a precursor in tetrapyrrole biosynthesis, under light illumination and upon addition of levulinic acid as an inhibitor of ALA dehydratase. To generate an industrial strain which produces ALA in the absence of light, we sequentially mutated R. sphaeroides CR-286 using N-methyl-N'-nitro-N-nitrosoguanidine (NTG). The mutant strains were screened by cultivating in the absence of light and assayed for ALA by the Ehrlich reaction in a 96-well microtiter plate. The mutant strain CR-386, derived from R. sphaeroides CR-286, was selected as a mutant that exhibited significant ALA accumulation. While CR-286 required light illumination for ALA production, CR-386 was able to accumulate 1.5 mM ALA in the presence of 50 mM glucose, 60 mM glycine, 15 mM levulinic acid and 1.0% (w/v) yeast extract under conditions of agitation in the absence of light. The mutant strain CR-450, derived from strain CR-386, was selected further as a mutant that exhibited significant ALA accumulation but no accumulation of aminoacetone, analogue of ALA. CR-450 accumulated 3.8 mM ALA under the same conditions. In the presence of 50 mM glucose, 60 mM glycine, 5 mM levulinic acid and 1.0% (w/v) yeast extract, the mutant strain CR-520, derived from strain CR-450, and strain CR-606, derived from strain CR-520, accumulated 8.1 mM and 11.2 mM ALA, respectively. In batch fermentation, the strain CR-606 accumulated 20 mM ALA over 18 h after the addition of glycine, levulinic acid, glucose and yeast extract.     

Impact of trace element addition on degradation efficiency of volatile fatty acids, oleic acid and phenyl acetate and on microbial populations in a biogas digester

The effect of trace element addition on anaerobic digestion of food industry- and household waste was studied using two semi-continuous lab-scale reactors, one (R30+) was supplied with Fe, Co and Ni, while the other (R30) acted as a control. Tracer analysis illustrated that methane production from acetate proceeded through syntrophic acetate oxidation (SAO) in both digesters. The effect of the trace elements was also evaluated in batch assays to determine the capacity of the microorganisms of the two digesters to degrade acetate, phenyl acetate, oleic acid or propionate, butyrate and valerate provided as a cocktail. The trace elements addition improved the performance of the process giving higher methane yields during start-up and early operation and lower levels of mainly acetate and propionate in the R30+ reactor. The batch assay showed that material from R30+ gave effects on methane production from all substrates tested. Phenyl acetate was observed to inhibit methane formation in the R30 but not in the R30+ assay. A real-time PCR analysis targeting methanogens on the order level as well as three SAO bacteria showed an increase in Methanosarcinales in the R30+ reactor over time, even though SAO continuously was the dominating pathway for methane production. Possibly, this increase explains the low VFA-levels and higher degradation rates observed in the R30+ batch incubations. These results show that the added trace elements affected the ability of the microflora to degrade VFAs as well as oleic acid and phenyl acetate in a community, where acetate utilization is dominated by SAO.     

Evaluation of predictions of volatile fatty acid production rates by the Molly cow model

Predicting ruminal volatile fatty acid (VFA) production is important, as VFA are an energy source to the animal, affect nutrient partitioning, and dictate methane production. The VFA production submodel in the Molly cow model was evaluated using data from 8 publications that reported VFA production rates for cattle. Evaluations were conducted with ruminal water balance predictions enabled and the ruminal VFA stoichiometry coefficients set to “mixed” for all diets, or “mixed” when forage represented between 20 and 80% of the diet, “concentrate” when <20% forage, or “forage” when >80% forage. Prediction errors were relatively insensitive to changes in VFA coefficients by diet type. Root mean square prediction errors (RMSPE) were 63, 63, and 49% for acetate, propionate, and butyrate production rates, respectively. A large proportion of the error was slope bias for acetate and butyrate, and a modest proportion for propionate. Because interconversions between acetate and propionate represent approximately 15% of the variation in net production rates, lack of such consideration in the model may contribute to the substantial model prediction errors. The potential of using thermodynamic equations to predict interconversions was assessed using observed ruminal pH and VFA concentrations from 2 studies and assuming constant hydrogen pressure and concentrations of CO₂, H₂O, adenosine diphosphate, ATP, and inorganic P. Rate constants for conversion of acetate to propionate and propionate to acetate were derived independently from the control treatments and used to predict the fluxes for the other treatment. The observed changes in VFA concentrations and pH explained the observed changes in conversion of acetate to propionate, but overpredicted the change in the propionate to acetate flux in one study. When applied to the other study, the equations predicted the increase in propionate to acetate flux, but failed to predict the observed reduction in acetate to propionate flux. The inability to predict responses accurately may be due to a lack of data for controlling factors other than pH and VFA concentrations.     

Mass transfer of water and volatile fatty acids in cocoa beans during drying

In order to elucidate the phenomena involved in the remnant acidity of cocoa beans dried artificially, the diffusivities of water, and volatile fatty acids (VFAs: acetic, propionic, butyric and iso-butyric acids) in cocoa beans during drying were evaluated. Experimental drying kinetics of the acids were conduced at 40–60 °C with and without shell. Samples were taken at different drying times for moisture and acids content evaluation. VFAs content was evaluated by GC in a methanolic extract, and moisture content by a vacuum oven. Mass diffusivity was estimated from the fitting of experimental kinetics to a theoretical model that takes into consideration the beans’ shape. Acetic, propionic and butyric acids diffusivities were significantly (p < 0.05) smaller than water diffusivities both with and without shell. VFAs diffusivities were between 1/6 and 1/22 diffusivities values for water. Iso-butyric acid diffusivity was not statistically significant but the value was smaller than for the other VFAs. The diffusivities of VFAs may explain the remnant acidity in artificially dried cocoa beans.     

日龄对盐水鹅挥发性风味成分和脂肪酸的影响

以不同日龄盐水鹅腿肉为原料,采用气相色谱-质谱联用仪(gas chromatography-mass spectrometer,GC-MS)对鹅腿样进行挥发性风味成分和脂肪酸检测。结果表明,随日龄增长,盐水鹅挥发性风味成分中醛类、酯类、酸类、酮类、苯类、醚类化合物相对含量上升,萜烯烃类和呋喃类化合物相对含量下降;不饱和脂肪酸、多不饱和脂肪酸、必需脂肪酸的相对含量和多不饱和脂肪酸/饱和脂肪酸值(Polyunsaturated fatty acids/Saturated fatty acids,P/S值)降低;日龄为150d盐水鹅挥发性风味成分和脂肪酸种类均最高。选择150d日龄扬州大白鹅能够同时保证盐水鹅的风味和营养。     

Volatile compounds of fresh and dry-cured loin as affected by dietary conjugated linoleic acid and monounsaturated fatty acids

Three levels (0%, 1% and 2%) of an enriched conjugated linoleic acid oil (CLA) were combined with two levels of monounsaturated fatty acids (MUFA) (low (19%) average and high (39%) average) for pig feeding. The profile of volatile compounds of fresh and dry-cured loin as affected by dietary CLA, MUFA and CLA × MUFA interaction was studied by headspace-solid phase microextraction coupled to gas chromatography–mass spectrometry. A total of 27 and 69 compounds were identified in fresh loin and dry-cured loin, respectively. Identified compounds were alcohols, aldehydes, aliphatic hydrocarbons, aromatic hydrocarbons, esters, furans, ketones, nitrogen compounds, sulphur compounds and terpenes. No qualitative differences in volatile compounds caused by the assayed treatments were found neither in fresh loin nor in dry-cured loin. Dietary CLA, MUFA and their interaction did not affect the level of most detected volatiles. Nevertheless, in fresh loin, the level of heptanal significantly increased due to dietary CLA (p = 0.001) and the area units (AU) for ethyl benzene and 1,3-dimethyl benzene increased at 1% CLA (p < 0.05). In the case of dry-cured loin, the AU of heptanal, nonanal and 2-nonenal contents increased due to dietary CLA (p = 0.016, 0.024 and 0.019, respectively).     

Assessing milk response to different combinations of branched-chain volatile fatty acids and valerate in Jersey cows

Some cellulolytic bacteria require 1 or more branched-chain volatile fatty acids (BCVFA) for the synthesis of branched-chain AA and branched-chain long-chain fatty acids because they are not able to uptake branched-chain AA or lack 1 or more enzymes to synthesize branched-chain AA de novo. Supplemental BCVFA and valerate were included previously as a feed additive that was later removed from the market; these older studies and more current studies have noted improvements in neutral detergent fiber digestibility and milk efficiency. However, most studies provided a single BCVFA or else isobutyrate (IB), 2-methylbutyrate (MB), isovalerate, and valerate altogether without exploring optimal combinations. Our objective was to determine a combination of isoacids that is optimal for milk production. Sixty (28 primiparous and 32 multiparous) lactating Jersey cows (106 ± 54 days in milk) were blocked and assigned randomly to either a control (CON) treatment without any isoacids, MB [12.3 mmol/kg dry matter (DM)], MB + IB (7.7 and 12.6 mmol/kg DM of MB and IB, respectively), or all 4 isoacids (6.2, 7.3, 4.2, and 5.1 mmol/kg DM of MB, IB, isovalerate, and valerate, respectively). Cattle were fed the CON treatment for a 2-wk period, then were assigned randomly within a block to treatments for 8 wk (n = 15). There was a trend for an interaction of supplement and parity for milk components. There were no differences in components for primiparous cows, whereas MB + IB tended to increase protein concentration by 0.04 and 0.08 percentage units in multiparous cows compared with the CON and MB treatments, respectively. Feeding MB + IB increased fat concentration by 0.23 to 0.31 percentage units compared with all other treatments in multiparous cows. Milk yield and dry matter intake (DMI) did not change with treatment. Treatment interacted with week for milk net energy for lactation/DMI; MB + IB tended to increase milk net energy of lactation/DMI by 0.10 Mcal/kg compared with MB and approached a trend for CON, mainly during the early weeks of the treatment period, whereas differences decreased during the last 2 wk of the treatment period. Cows fed MB had the highest 15:0 anteiso fatty acids in the total milk fatty acid profile, which was greater than that for CON or MB + IB cows, but not cows supplemented with isoacids. Cows fed MB alone had the numerically lowest milk net energy for lactation/DMI. The combination of MB + IB appeared optimal for increasing feed efficiency in our study and was not at the expense of average daily gain. Further research is needed for evaluating how potential changes in supplemental isoacid dosage should vary under differing dietary conditions.     

Effect of forage:concentrate ratio on fatty acid composition of rumen bacteria isolated from ruminal and duodenal digesta

Four dairy cows were used to examine the effect of the dietary forage:concentrate ratio [35:65, 50:50, 65:35, and 80:20 on a dry matter (DM) basis] on the fatty acid composition of rumen bacteria isolated from the liquid (LAB) and solid (SAB) phase of the rumen and duodenal digesta. Rumen contents were sampled 4 h after the morning feeding. Solid and liquid phases were separated from rumen contents and duodenal bacteria from a composite duodenal sample by differential centrifugation. Total fatty acid content in bacterial DM was 1.6 to 2.8 times higher in SAB compared with LAB, and increased with dietary concentrate. In combination with published reports, the data show that bacterial fatty acid content and composition is closely related to dietary fatty acids except for C18:2n-6 and C18:3n-3. A decrease in forage:concentrate ratio increased bacterial concentration of trans-10 C18:1, and this increase was 3.4 times higher in LAB compared with SAB. Analysis of odd- and branched-chain fatty acids showed large differences between SAB and LAB, which probably reflected a difference in species composition. The variation in odd- and branched-chain fatty acids between SAB and LAB was used to estimate their relative proportions in duodenal bacteria by means of linear programming, and showed an increased proportion of SAB from 64.7 to 74.8% with increasing forage:concentrate ratio. In addition, increasing the proportion of dietary forage was closely related to the proportion of anteiso C15:0 in total odd- and branched-chain fatty acids (r_pearson = −0.771). The bacterial concentration of iso C17:0 closely reflected the bacterial growth rate as shown by the relation with cytosine:N (r_pearson = −0.729). These strong relationships suggest that odd- and branched-chain fatty acids might be used as tool to evaluate nutrient supply to rumen bacteria.     

Evaluation of volatile compounds and free fatty acids in set types yogurts made of ewes’, goats’ milk and their mixture using two different commercial starter cultures during refrigerated storage

Six different types of yogurt were manufactured from Damascus goat milk, Awassi ewe milk and a mixture of equal portions of the 2 species of milk using 2 types of commercial yogurt cultures (CH-1 and YF-3331). Yogurts were chemically analysed at 1, 7, 14 and 21 days of storage. Results showed that cultures significantly affected acetaldehyde (P < 0.05), acetone (P < 0.05) and diacetyl (P < 0.001) contents. Type of milk significantly influenced acetaldehyde (P < 0.05), diacetyl (P < 0.001), acetoin (P < 0.001) and ethanol (P < 0.05) levels. Significant variations occurred in acetaldehyde (P < 0.001) and acetoin (P < 0.05) contents during the storage. Short-chain free fatty acids were the highest in ewes’ milk yogurt made with culture YF-3331, and increased during storage, while the levels of medium-chain free fatty acids, except for decanoic acid, were unchanged and the amount of long-chain free fatty acids decreased during storage. Cultures used and types of milk had no effect on long-chain free fatty acids in yogurts.     

不同部位牛油脂肪酸组成和挥发性风味成分分析

以1 000 kg牛脂肪为每个样品量提炼牛油,对得到的肾周脂、腹脂和分割脂3种不同部位的牛油采用气相色谱进行脂肪酸组成分析,并使用气相色谱–嗅闻–质谱联用进行了挥发性风味成分检测。结果表明：3种牛油的饱和脂肪酸以棕榈酸、硬脂酸为主,不饱和脂肪酸以油酸为主。3种牛油的棕榈酸含量无显著差异（P>0.05）,腰油与肚油的硬脂酸、油酸含量无显著差异（P>0.05）,而分割油与腰油、肚油的硬脂酸、油酸含量都具有极显著性差异（P<0.01）。3种牛油中挥发性风味成分主要有醛、酸、醇、酮、酯和杂环6大类,其中醛类与酸类含量之和可占总量70%以上。不同部位牛油之间挥发性风味成分含量具有显著差异（P<0.05）。腰油中挥发性风味物质总量最高,肚油中醛类含量占比达到59.90%;分割油中挥发性风味成分种类最多,其中酯类、杂环类挥发性风味成分占比分别达到8.49%与7.61%。脂肪酸测定和挥发性风味成分分析揭示了不同部位牛油之间的特征与差异,可为食用牛油产品的合理开发利用和风味品质控制提供参考数据。     

Influence of dietary conjugated linoleic acid on the fatty acid composition and volatile compounds profile of heavy pig loin muscle

This study evaluated the effects of dietary conjugated linoleic acid (CLA) on fatty acid composition, chemical composition and volatile compounds profile of the longissimus dorsi muscle in Italian heavy pigs. The animals (97 kg) were randomly assigned to three diets varying in supplemental CLA (CON = 0 CLA, T1 = 2.5 g CLA kg⁻¹ feed and T2 = 5.0 g CLA kg⁻¹ feed) till the slaughtering at 172 kg. Samples of longissimus dorsi were analysed for chemical composition (moisture, protein and lipid content), fatty acid composition and volatile compounds. No significant differences were observed for proximate chemical composition. Dietary CLA showed limited effects on fatty acid composition of longissimus dorsi, with higher, but not significantly, amounts of saturated fatty acids in the treated groups than in the control group; both the cis‐9, trans‐11 and the trans‐10, cis‐12 isomers of CLA were increased in longissimus dorsi from pigs fed CLA. T1 and T2 pigs had a greater concentration of C16:0 and of C16:1 (P < 0.01) than CON. CLA diets tended to reduce C20:2 (P = 0.077) and C20:4 (P = 0.065) content in longissimus dorsi muscle. Diets containing higher amount of CLA were responsible for increased levels of volatile compounds in meat, but not at a significant level. Copyright © 2005 Society of Chemical Industry     

光强和温度对球等鞭金藻（Isochrysis sphaerica）生长及其脂肪酸的影响

本研究测定了光照强度为16～58μmol／（m^2·s）和15～30℃条件下球等鞭金藻（Isochrysissphaerica）的生长曲线以及脂肪酸组成和含量。结果表明,在设置的光照强度范围内,光照强度越强,微藻的生长速度越快。光强对各种脂肪酸的含量影响有差异,过低和过高都不利于藻体内多不饱和脂肪酸（PolyunsaturatedFattyAcids,PU—FAs）的积累。当光强为24μmol／（m^2·s）时二十二碳六烯酸（DoeosahexaenoicAcid,DHA,C22：6n～3）的积累量最高,占总脂肪酸的11．77％。球等鞭金藻的最适生长温度在20～25℃之间,低温条件有利于不饱和脂肪酸的积累。光强20μmol／（m^2·s）、温度20℃条件下总不饱和脂肪酸（TotalUnsaturatedFattyAcids,TUFAs）含量最高,达64．99％;同样的光强,温度15℃条件下DHA占总脂肪酸的12．19％。可见适宜的光强和温度是利用球等鞭金藻收获PUFAs的关键。     

Stability of fatty acid composition after thermal,high pressure,and microwave processing of cow milk as affected by polyunsaturated fatty acid concentration

Interest has been increasing to enhance the contents of healthy polyunsaturated fatty acid (PUFA) in milk. However, trans fatty acids and conjugated linoleic acid (CLA) can be altered after thermal processing and high pressures disrupt the milk fat globule membrane, exposing the lipid core and helping its oxidation. The objective of the present research was to study whether processing can alter the fatty acid composition of milk and if these changes are affected by PUFA concentration as previous studies suggest. Two cow milk batches (500 L each), one naturally enriched in PUFA, were processed to obtain pasteurized; high temperature, short time; UHT; high pressure; and microwave pasteurized samples. The detailed fatty acid composition was analyzed with special attention to trans fatty acids and CLA isomers. Results showed that after high temperature, short time processing, total CLA content increased in both milk batches, whereas sterilization resulted in a sigmatropic rearrangement of C18:2 cis-9,trans-11 to C18:2 trans-9,trans-11. The extent of these effects was greater in milks naturally enriched in PUFA.     

Oxidative changes of fresh loin from pig,caused by dietary conjugated linoleic acid and monounsaturated fatty acids,during refrigerated storage

Three levels (0%, 1% and 2%) of a conjugated linoleic acid enriched oil (CLA) were combined with two levels of monounsaturated fatty acids (MUFA) (19% and 39% average) for pig feeding. Changes in instrumental colour, lipid oxidation (thiobarbituric acid reactive substances, TBARS) and volatile profile of fresh loin chops, as affected by dietary CLA, MUFA and CLA × MUFA, were studied throughout 7 days of refrigerated storage. Lightness (L∗) evolution was conditioned by dietary CLA, whereas changes in redness (a∗) and yellowness (b∗) were unaffected by dietary supplements. Dietary CLA at 2% led to higher TBARS values of loin chops at day 7 of refrigerated storage (p < 0.05), while MUFA supplementation and CLA × MUFA interaction did not affect lipid oxidation. Dietary CLA, MUFA or CLA × MUFA did not affect most volatile compounds of loin chops after 7 days of storage.     

接种乳酸菌对腌干鱼总脂肪及游离脂肪酸的影响

在低盐腌制的鱼肉中接种复合乳酸菌,发酵及干燥制成腌干鱼。研究接种复合乳酸菌对腌干鱼总脂肪和游离脂肪酸的影响,结果表明:接种乳酸菌腌制的鱼肉(EG)总脂肪相对含量低于非接种组(CK),游离脂肪酸总量高于非接种组。EG组鱼的不同游离脂肪酸含量高低顺序为:多不饱和脂肪酸>单不饱和脂肪酸>饱和脂肪酸。CK组鱼的不同游离脂肪酸含量高低顺序为:多不饱和脂肪酸>饱和脂肪酸>单不饱和脂肪酸。其游离脂肪酸变化与非接种组比较,表现为饱和脂肪酸含量显著减少,不饱和脂肪酸含量显著增加。其饱和脂肪酸含量显著减少的有:棕榈油酸(C16∶0)和十八酸(C18∶0),不饱和脂肪酸显著增加的有:花生四烯酸(C20∶4)和二十碳五烯酸(C20∶5)。接种乳酸菌在一定程度上提高了腌干鱼肉的营养价值和食用价值。      

Effect of dietary conjugated linoleic acid in combination with monounsaturated fatty acids on the composition and quality traits of cooked loin

Three levels (0%, 1% and 2%) of a conjugated linoleic acid oil (CLA) were combined with two levels of monounsaturated fatty acids (MUFA) (low – 19% average and high – 39% average) for pig feeding (n = 48, eight animals per treatment). The composition and quality traits (fat content, cooking losses, lipid oxidation, fatty acid profile, volatile profile and sensory analysis) of cooked loin, as affected by dietary CLA, MUFA, and CLA × MUFA interaction were studied. CLA and CLA × MUFA did not affect the intramuscular fat content, cooking losses, lipid oxidation, volatile profile and sensory traits of cooked meat. Therefore, CLA could be supplemented to the pig diet without detriment of the measured quality traits of cooked meat, and regardless of the MUFA level of pig diets. Dietary CLA increased the content of SFA and decreased the level of MUFA of meat, and led to a CLA enrichment, regardless the MUFA level of pig diets, but the content of CLA isomers of fresh meat decreased after the cooking process.     

武陵青花椒壳挥发油成分及脂肪酸分析

分别采用水蒸气蒸馏法和同时蒸馏萃取法提取武陵青花椒壳中的挥发油成分,提取率分别为6.4%和8.5%。再用气相色谱-质谱法（GC-MS）对挥发油成分进行分析,用峰面积归一化法确定各组分的相对含量。两种方法均鉴定出43种化合物,其中水蒸气蒸馏法以β-芳樟醇（66.91%）、反式-橙花叔醇（6.28%）、D-柠檬烯（5.24%）、4-萜烯醇（2.6%）为主,同时蒸馏萃取法以β-芳樟醇（64.32%）、D-柠檬烯（5.97%）、反式-橙花叔醇（4.17%）、反式-9-十八烯酸（3.09%）为主。同时对挥发油进行甲酯化,然后用GC-MS对挥发油中含有的脂肪酸进行了分析,共分离鉴定出6种脂肪酸。结果表明:两种方法提取的挥发油的组成及含量存在差异,挥发油中主要为不饱和脂肪酸。水蒸气蒸馏法对无极性和弱极性的萜烯和萜醇类化合物提取效果较好,而同时蒸馏萃取法适于各种极性化合物,对极性稍强的脂肪酸和酯类有较好的提取效果。      

Response of forage fiber degradation by ruminal microorganisms to branched-chain volatile fatty acids,amino acids,and dipeptides

This study evaluated the effect of branched-chain volatile fatty acids (VFA; isobutyric acid, isovaleric acid), amino acids (valine, leucine), and dipeptides (valine-valine, leucine-leucine) on neutral detergent fiber (NDF) degradation by rumen microorganisms in vitro. The CP (%) and in situ NDF degradation rate (%/h) for alfalfa, bermudagrass, and pangolagrass hays, and napiergrass silage were 17.2 and 7.5, 4.7 and 3.1, 8.3 and 5.3, and 9.6 and 3.4, respectively. In vitro NDF digestibility was the lowest for bermudagrass; alfalfa and napiergrass were the highest. When the incubation contained more ammonia initially, digestibilities increased, but relative differences among forages were unchanged. Adding branched-chain VFA (2 mM) to incubations increased digestibilities more than controls on 15 out of 16 occasions. The effectiveness varied with isoacids and forages used. Amino acid (2 mM) or dipeptide (1 mM) addition consistently increased digestibility over controls. Amino acids further increased digestibility over corresponding isoacids on 14 occasions. Improvement in digestibility over control by leucine appeared to be greater than that by valine. Digestibilities with dipeptides were always greater than those with isoacids, except for one case. Dipeptide addition further increased digestibility significantly over corresponding amino acids on only six occasions, while percent improvement in digestibility numerically by dipeptides occurred in 10 cases. Valine-valine seemed to exert different effect than leucine-leucine, depending on initial ammonia availability. The results indicate that dipeptides could be more effective than isoacids and amino acids in improving NDF digestion. Forages with high CP content or rapid NDF degradation rate appeared to respond to additives to smaller degrees.