• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.739-747
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• 2018

The low solubility and high-cost recovery of Propionibacterium acnes polyunsaturated fatty acid isomerase (PAI) are key problems in the bioproduction of high value-added conjugated linoleic acid (CLA). To improve the solubility of recombinant PAI, six chaperone proteins were coexpressed with PAI. Introduction of GroELS proteins dramatically improved the PAI solubility from 29% to 97%, with increased activity by 57.8%. Combined expression of DnaKJ-GrpE and GroELS proteins increased the activity by 11.9%. In contrast, coexpression of DnaKJ-GrpE proteins significantly reduced the activity by 57.4%. Plasmids pTf16 harboring the tig gene and pG-Tf2 containing the tig and groEL-groES genes had no visible impact on PAI expression. The lytic protein E was then introduced into the recombinant Escherichia coli to develop a cell autolysis system. A 35% activity of total intracellular PAI was released from the cytoplasm by suspending the lysed cells in distilled water. The PAI recovery was further improved to 81% by optimizing the release conditions. The lipase from Rhizopus oryzae was also expressed in E. coli, with an extracellular activity of 110.9 U/ml. By using the free PAI and lipase as catalysts, a joint system was established for producing CLA from sunflower oil. Under the optimized conditions, the maximum titer of t-10,c-12-CLA reached 9.4 g/l. This work provides an effective and low-cost strategy to improve the solubility and recovery of the recombinant intracellular PAI for further large-scale production of CLA.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.11
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• pp.1655-1661
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• 2005

The present studies were designed to provide new information on fatty acid profiles of various muscles and adipose tissues of fattening horses in comparison with beef cattle and pigs. In the first study, the lipids were extracted respectively from subcutaneous, intermuscular adipose tissues, longissimus dorsi and biceps femoris muscles of fattening Breton horses (n = 8) with an average body weight of 1,124 kg. In the second study, the lipids were extracted from subcutaneous, intermuscular adipose tissues and longissimus dorsi muscle of fattening horses (n = 13), Japanese Black beef cattle (n = 5), Holstein steers (n = 5) and fattening pigs (n = 5). The fatty acids in the lipid samples were determined by gas chromatography after methylation by a combined base/acid methylation method. It was found that the lipids from horse subcutaneous and intermuscular adipose tissues contained more (p<0.05) polyunsaturated fatty acids (PUFA) which were mainly composed of linoleic acid (C18:2) and linolenic acid (C18:3) than those in the muscles. The weight percent of conjugated linoleic acids (CLA cis 9, trans 11) in lipids from biceps femoris muscle was 0.22%, which was higher (p<0.05) than that from the other depots. The horse lipids were higher (p<0.05) in PUFA but lower (p<0.05) in SFA and MUFA in comparison with those of the cattle and pigs. The percentage of C18:2 or C18:3 fatty acid in the horse lipids were respectively 2-8 fold or 5-18 fold higher (p<0.05) than those of the cattle and pigs. The percentages of CLA (cis 9, trans 11) in the horse lipids (0.14-0.16%) were very close to those of the pigs (0.18-0.19%) but much lower (p<0.05) than those of the Japanese Black beef cattle (0.55-0.94%) and Holstein steers (0.46-0.71%). The results indicated that the fatty acid profiles of lipids from different muscle and adipose tissues of fattening horses differed significantly. In comparison with that of the beef cattle and pigs, the horse lipids contained more C18:2 and C18:3 but less CLA.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.10
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• pp.1364-1373
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• 2012

Four crossbred (75% Holstein Friesian) lactating dairy cows, with an average live weight of $418{\pm}5$ kg and $36{\pm}10$ d in milk were randomly assigned according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design to evaluate the effects of cassava hay (CH) and rice bran oil (RBO) on feed intake, nutrient digestibility, ruminal fermentation, milk yield, and milk composition. Factor A was non-supplementation or supplementation with CH in the concentrate. Factor B was supplementation with RBO at 0% or 4% in the concentrate mixture. The four dietary treatments were (T1) control (Concentrate with non-CH plus 0% RBO; C), (T2) Concentrate with CH plus 0% RBO (CH), (T3) Concentrate with non-CH plus 4% RBO (RBO), and (T4) Concentrate with CH plus 4% RBO (CHRBO). The cows were offered concentrate, at a ratio of concentrate to milk production of 1:2, and urea-lime treated rice straw was fed ad libitum. Urea-lime treated rice straw involved 2.5 g urea and 2.5 g $Ca(OH)_2$ (purchased as hydrated lime) in 100 ml water, the relevant volume of solution was sprayed onto a 100 g air-dry (91% DM) straw, and then covering the stack with a plastic sheet for a minimum of 10 d before feeding directly to animals. The CH based concentrate resulted in significantly higher roughage intake and total DM intake expressed as a percentage of BW (p<0.05). Ruminal pH, $NH_3$-N, BUN and total VFA did not differ among treatments, while RBO supplementation increased propionate, but decreased acetate concentration (p<0.05). Furthermore, the population of total ruminal bacteria was significantly lower on the RBO diet (p<0.05). In contrast, the total ruminal bacteria and cellulolytic bacteria on the CH diet were higher than on the other treatments. Supplementation with CH increased (p<0.05) F. succinogens and R. flavefaciens populations, whereas the populations of B. fibrisolvens and M. elsdenii were increased on the RBO diet. In addition, supplementation with CH and RBO had no effect on milk production and composition in dairy cows, while fatty acid composition of milk was influenced by RBO supplementation, and resulted in significantly lower (p<0.05) concentrations of both short-chain and medium-chain FA, and increased (p<0.05) the proportion of long-chain FA in milk fat, as well as significantly increased cis-9, trans-11 CLA and total CLA. In conclusion, RBO or CH exhibited specific effects on DMI, rumen fermentation, microbial population, milk yield and composition in lactating dairy cows, which were not interactions between CH and RBO in the diets. Feeding lactating dairy cows with RBO could improve fatty acid in milk fat by increasing cis-9, trans-11 CLA.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.7
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• pp.951-959
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• 2014

The objective of this study was to determine the effects of linseed oil or whole linseed supplementation on performance and milk fatty acid composition of lactating dairy cows. Thirty six Holstein Friesian crossbred lactating dairy cows were blocked by milking days first and then stratified random balanced for milk yields and body weight into three groups of 12 cows each. The treatments consisted of basal ration (53:47; forage:concentrate ratio, on a dry matter [DM] basis, respectively) supplemented with 300 g/d of palm oil as a positive control diet (PO), or supplemented with 300 g/d of linseed oil (LSO), or supplemented with 688 g/d of top-dressed whole linseed (WLS). All cows were received ad libitum grass silage and individually fed according to the treatments. The experiment lasted for 10 weeks including the first 2 weeks as the adjustment period, followed by 8 weeks of measurement period. The results showed that LSO and WLS supplementation had no effects on total dry matter intake, milk yield, milk composition, and live weight change; however, the animals fed WLS had higher crude protein (CP) intake than those fed PO and LSO (p<0.05). To compare with the control diet, dairy cow's diets supplemented with LSO and WLS significantly increased milk concentrations of cis-9,trans-11-conjugated linoleic acid (CLA) (p<0.05) and n-3 fatty acids (FA) (p<0.01), particularly, cis-9,12,15-C18:3, C20:5n-3 and C22:6n-3. Supplementing LSO and WLS induced a reduction of medium chain FA, especially, C12:0-C16:0 FA (p<0.05) while increasing the concentration of milk unsaturated fatty acids (UFA) (p<0.05). Milk FA proportions of n-3 FA remarkably increased whereas the ratio of n-6 to n-3 decreased in the cows supplemented with WLS as compared with those fed the control diet and LSO (p<0.01). In conclusion, supplementing dairy cows' diet based on grass silage with WLS had no effect on milk yield and milk composition; however, trans-9-C18:1, cis-9,trans-11-CLA, n-3 FA and UFA were increased while saturated FA were decreased by WLS supplementation. Therefore, it is recommended that the addition 300 g/d of oil from whole linseed should be used to lactating dairy cows' diets.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.2
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• pp.234-239
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• 2012

The objective of this study was to evaluate the effects of forage level and oil supplement on selected strains of rumen bacteria believed to be involved in biohydrogenation (BH). A continuous culture system consisting of four fermenters was used in a $4{\times}4$ Latin square design with a factorial arrangement of treatments, with four 10 d consecutive periods. Treatment diets were: i) high forage diet (70:30 forage to concentrate (dry matter basis); HFC), ii) high forage plus oil supplement (HFO), iii) low forage diet (30:70 forage to concentrate; LFC), and iv) low forage plus oil supplement (LFO). The oil supplement was a blend of fish oil and soybean oil added at 1 and 2 g/100 g dry matter, respectively. Treatment diets were fed for 10 days and samples were collected from each fermenter on the last day of each period 3 h post morning feeding. The concentrations of vaccenic acid (t11C18:1; VA) and c9t11 conjugated linoleic acid (CLA) were greater with the high forage diet while the concentrations of t10 C18:1 and t10c12 CLA were greater with the low forage diet and addition of oil supplement increased their concentrations at both forage levels. The DNA abundance of Anaerovibrio lipolytica, and Butyrivibrio fibrisolvens vaccenic acid subgroup (Butyrivibrio VA) were lower with the low forage diets but not affected by oil supplement. The DNA abundance of Butyrivibrio fibrisolvens stearic acid producer subgroup (Butyrivibrio SA) was not affected by forage level or oil supplement. In conclusion, oil supplement had no effects on the tested rumen bacteria and forage level affected Anaerovibrio lipolytica and Butyrivibrio VA.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.2
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• pp.204-213
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• 2008

Four ruminally cannulated Holstein cows averaging 43 days in milk (DIM) were used in a $4{\times}4$ Latin square to determine the effect of feeding extruded soybean, ground canola seed and whole cottonseed on ruminal fermentation and milk fatty acid profile. One hundred and twenty lactating Holstein cows, 58 (${\pm}31$) DIM, were assigned to four treatments in a completely randomized block design to study the effects of the three types of oilseeds on production parameters and milk fatty acid profile. The four diets were a control diet (CON) and three diets in which 10% extruded soybean (ESB), 5% ground canola seed (GCS) and 10% whole cottonseed (WCS) were included, respectively. Diets consisted of concentrate mix, corn silage and Chinese wild rye and were balanced to similar concentrations of CP, NDF and ADF. Ruminal fermentation results showed that ruminal fermentation parameters, dry matter intake and milk yield were not significantly affected by treatments. However, compared with the control, feeding cows with the three oilseed diets reduced C14:0 and C16:0 and elevated C18:0 and C18:1 concentrations in milk, and feeding ESB increased C18:2 and cis9, trans11 conjugated linoleic acid (CLA). Production results showed that feeding ESB tended to increase actual milk yield (30.85 kg/d vs. 29.29 kg/d) and significantly decreased milk fat percentage (3.53% vs. 4.06%) compared with CON. Milk protein (3.41%) and solid non-fat (13.27%) from cows fed WCS were significantly higher than from cows fed CON (3.24% and 12.63%, respectively). Milk urea N concentrations from cows fed the ESB (164.12 mg/L) and GCS (169.91 mg/L) were higher than cows fed CON (132.31 mg/L). However, intake of DM, 4% fat corrected milk, energy corrected milk, milk fat and protein yields, milk lactose percentage and yield, somatic cell count and body condition score were not affected by different treatments. The proportion of medium-chain fatty acid with 14 to 16 C units in milk was greatly decreased in cows fed ESB, GCS and WCS. Feeding ESB increased the concentration in milk of C18:1, C18:2, C18:3 and cis9, trans11-CLA content by 16.67%, 37.36%, 95.24%, 72.22%, respectively, feeding GCS improved C18:0 and C18:1 by 17.41% and 33.28%, respectively, and feeding WCS increased C18:0 by 31.01% compared with feeding CON. Both ruminal fermentation and production trial results indicated that supplementation of extruded soybean, ground canola seed and whole cottonseed could elevate the desirable poly- and monounsaturated fatty acid and decrease the medium chain fatty acid and saturated fatty acid content of milk fat without negative effects on ruminal fermentation and lactation performance.

• Journal of Animal Science and Technology
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• v.57 no.12
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• pp.42.1-42.14
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• 2015

Background: Dietary fats can alter the deposition and distribution of body fats in ruminants. The deposition and distribution of body fat play a vital role in the quality of ruminant carcasses and are of great commercial value since they influence the profitability and consumer acceptability of ruminant meat. The current study examined the effects of dietary blend of 80 % canola oil and 20 % palm oil (BCPO) on carcass characteristics, meat yield and accretion of fatty acid (FA) in subcutaneous, omental, perirenal, and mesentery adipose depots and m. supraspinatus (SS) in goats. Methods: Twenty four Boer crossbred bucks (BW $20.54{\pm}0.47kg$) were randomly assigned to diets containing on DM basis 0, 4 and 8 % BCPO, fed for 100 d and harvested. Results: Diet had no effect (P > 0.05) on slaughter weight, dressing percentage, carcass and non-carcass components, meat yield, color, moisture and carotenoid contents and weight of adipose tissues in goats. The proportion of C18:1n-9 and cis-9 trans-11 CLA in the omental, perirenal and SS was higher (P < 0.05) in goats fed 4 and 8 % BCPO compared with the control goats. Dietary BCPO reduced (P < 0.05) the proportion of C14:0 in the omental, perirenal and mesentery depots, C18:0 in the perirenal depot, C16:0 in the SS and C16:1n-7 in the SS, omental and perirenal tissues. Dietary BCPO enhanced the proportion of C18:1 trans-11 Vaccenic and C18:3n-3 in SS and C20:5n-3 in SS and mesentery depot. No significant changes were found in the FA composition of subcutaneous depot. Conclusions: Results indicate that dietary BCPO can be utilized to alter the FA composition of adipose tissues without detrimental effects on carcass characteristics in goats. Nonetheless, dietary BCPO is not an effective repartitioning agent for body fats in goats.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.12
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• pp.1712-1720
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• 2012

We investigated the effect of the amount of body condition loss in the dry period and early lactation in 42 high-yielding Holstein-Friesian cows on milk yield and the share of fatty acids in milk fat. Energy reserves were estimated based on the body condition scoring (BCS) and backfat thickness (BFT). Milk yield and milk composition were determined over 305-d lactation. From d 6 to 60 of lactation, the concentrations of 43 fatty acids in milk fat were determined by gas chromatography. Cows were categorized based on body condition loss from the beginning of the dry period to the lowest point of the BCS curve in early lactation into three groups: low condition loss group (L) ${\leq}0.5$ points (n = 14); moderate condition loss group (M) 0.75 to 1.0 points (n = 16) and high condition loss group (H) >1.0 points (n = 12). Cows whose body energy reserves were mobilized at 0.8 BCS and 11 mm BFT, produced 12,987 kg ECM over 305-d lactation, i.e. 1,429 kg ECM more than cows whose BCS and BFT decreased by 0.3 and 5 mm, respectively. In group H, milk yield reached 12,818 kg ECM at body fat reserve mobilization of 1.3 BCS and 17 mm BFT. High mobilization of body fat reserves led to a significant (approx. 5%) increase in the concentrations of monounsaturated fatty acids-MUFA (mostly $C_{18:1}$ cis-9, followed by $C_{18:1}$ trans-11), a significant decrease in the levels of fatty acids adversely affecting human health, and a drop in the content of linoleic acid, arachidonic acid and docosahexaenoic acid in milk fat. In successive weeks of lactation, an improved energy balance contributed to a decrease in the concentrations of unsaturated fatty acids (UFA) and an increase in the conjugated linoleic acid (CLA) content of milk fat.

• Journal of Animal Science and Technology
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• v.46 no.3
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• pp.373-386
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• 2004

This study was conducted to investigate effects of the brown seaweed residues supplementation on in vitro fermentation, and milk yield and milk composition of dairy cows. Therefore, two experiments consisting of an in vitro and an in vivo growth trial were used. In in vitro experiment, brown seaweed residues(BSR) was supplemented in basal diet with 0, 1, 2 and 4% respectively, and incubated for 3, 6, 9, 12, and 24 h. The pH value, ammonia-N and VFA were investigated. The pH value tended to increase with increasing BSR during the incubation. Particularly, pH was significantly higher in BSR treatments compared with control at 9 h(p < 0.05). While, ammonia-N concentration was not significantly different across treatments during the whole incubation. BSR supplementation did not affect total VFA production, but acetate was linearly increased in BSR treatments compared with control at 12 h(p < 0.05), and its concentration was highest(92.70 mM) in 4% BSR among treatments. The concentration of iso-butyrate tended to increase in BSR treatments in comparison to control during the incubation. In addition, the concentration of iso-valerate was higher in BSR treatments compared with control at 12 and 24 h. In growth trial, BSR was added(800 g/d/animaI) to diets of dairy cow. Dry matter intake was not affected by BSR supplementation, but daily milk yield(kg) significantly increased in BSR treatment compared with control(p < 0.05). However, milk composition(%) and milk yield(kg) were not significantly different between treatments. Milk fat(% and kg/d) tended to slightly decrease in BSR treatment compared with control(3.59% and 1.06 kg/d vs. 3.32% and 1.01 kg/d), The contents of C16:0 and C20:4 in milk significantly increased in BSR treatment compared with control reflecting from dietary fatty acid composition. The content of C18:0 in milk which is end product of biohydrogenation of CI8 unsaturated fatty acids in the rumen significantly increased in BSR treatment compared with control(p < 0.05). C18:2 content in milk tended to decrease, but tended to increase trans-II C18:l and CLA contents in milk in BSR treatment compared with control. In conclusion, it could be summarized that BSR may stabilize rumen pH, and it could improve milk yield and CIA content in milk with more than 4% of diet. Therefore, BSR could be beneficially used in dairy diets as a feed additive.