Effects of the essential oil cyclodextrin complexes on ruminal methane production in vitro

The effects of including essential oil cyclodextrin (CD) complexes on ruminal methane and hydrogen production, volatile fatty acid (VFA) and protozoa were studied in vitro. The essential oil CD complexes used in the present study were both αCD and βCD with cineol, eucalyptus, menthol, peppermint, thyme and wasabi. Diluted rumen fluid (60 mL) was incubated anaerobically at 38°C for 6 h with the essential oil CD complex (1–40 mg as oil). Eucalyptus‐αCD reduced methane production by 40% with 10 mg as oil along with decreasing the protozoal number and increasing total VFA and proportion of propionic acid. Both wasabi‐αCD and wasabi‐βCD reduced methane production by 85% and 97%, respectively, with 10 mg as oil, though they increased hydrogen production notably. The protozoal number was unchanged by wasabi‐βCD and decreased to 50% by wasabi‐αCD. Propionic acid was increased by both wasabi‐αCD and wasabi‐βCD. The other essential oil‐CD showed no significant effect on reducing methane production. The combined use of 20 mM of fumaric acid with wasabi‐αCD (1 mg as oil) caused about a 50% depression of methane production compared to the control without increasing hydrogen production. Fumaric acid was more effective than malic acid in reducing methane production. The addition of eucalyptus‐αCD, wasabi‐αCD or wasabi‐βCD together with organic acid to rumen fluid turned out to be effective for reducing methane production with no increase in hydrogen production.     

体外法研究硝酸钠调控水牛瘤胃甲烷生成对脂肪酸生物氢化途径的影响

本试验旨在研究硝酸钠调控水牛瘤胃甲烷生成对脂肪酸生物氢化途径的影响。选取3头体重约为（650±50）kg、安装永久性瘤胃瘘管的水牛作为瘤胃液供体动物,通过体外批次培养,设计发酵底物的精粗比为40：60,试验设4个组,每组5个重复,每组各添加0.25 mg/mL的α-亚麻酸,硝酸钠添加水平分别为0（对照）、1、2、3 mg/mL。分别培养3、6、9、12、24 h时测定产气量和甲烷产量,在培养24 h结束后测定体外发酵参数和脂肪酸含量。结果表明：①添加硝酸钠显著降低了瘤胃培养液24 h的总产气量、甲烷（CH₄）含量和甲烷/总产气量的比例（P<0.05）,添加1、2和3 mg/mL硝酸钠后瘤胃液甲烷含量分别降低了89.62%、91.20%、91.75%。②添加硝酸钠组瘤胃培养液的pH和氨态氮（NH₃-N）含量显著高于对照组（P<0.05）,添加1 mg/mL硝酸钠组微生物蛋白（MCP）含量也显著高于对照组（P<0.05）,其他组别差异不显著（P>0.05）;添加硝酸钠组瘤胃液乙酸浓度差异不显著（P>0.05）,但丙酸、丁酸、异丁酸、戊酸和异戊酸浓度显著低于对照组（P<0.05）,乙酸/丙酸显著高于对照组（P<0.05）,添加3 mg/mL硝酸钠组瘤胃液总挥发性脂肪酸（TVFA）含量显著低于对照组（P<0.05）。③添加1 mg/mL硝酸钠组瘤胃液C18：2 cis-9,trans-11、C18：2 trans-10,cis-12、C20：1、不饱和脂肪酸（UFA）含量及UFA/SFA显著高于其他组（P<0.05）;添加硝酸钠组瘤胃液C18：2n6c、C18：1n9t、C20：5n3（EPA）和C22：6n3（DHA）的含量均高于对照组,且1 mg/mL硝酸钠组含量最高,但差异不显著（P>0.05）;添加1 mg/mL硝酸钠组瘤胃液C18：3n3、C18：2n6c和C18：1n9c含量均高于对照组,差异不显著（P>0.05）。由此可见,体外添加硝酸钠显著降低了瘤胃液总产气量和甲烷含量,pH和NH₃-N含量显著升高,TVFA含量降低,通过显著减少丙酸含量而升高乙酸/丙酸;添加1 mg/mL硝酸钠可显著提高瘤胃液共轭亚油酸（CLA）和UFA含量,且在抑制甲烷产生的同时能够降低不饱和脂肪酸的生物氢化程度。     

Genome‐wide association study for fatty acid composition in Japanese Black cattle

Fatty acid composition is one of the important traits in beef. The aim of this study was to identify candidate genomic regions for fatty acid composition by genome‐wide association study with 50 K single nucleotide polymorphism (SNP) array in Japanese Black cattle. A total of 461 individuals and 40 657 SNPs were used in this study. We applied genome‐wide rapid association using mixed model and regression (GRAMMAR) and genomic control approaches to estimate the associations between genotypes and fatty acid composition. In addition, two SNPs in fatty acid synthase (FASN) (T1952A) and stearoyl‐CoA desaturase (SCD) (V293A) genes were also genotyped. Association analysis revealed that 30 significant SNPs for several fatty acids (C14:0, C14:1, C16:1 and C18:1) were located in the BTA19 FASN gene located within this region but the FASN mutation had no significant effect on any traits. We also detected one significant SNP for C18:1 on BTA23 and two SNPs for C16:0 on BTA25. The region around 17 Mb on BTA26 harbored two significant SNPs for C14:1 and SNP in SCD in this region showed the strongest association with C14:1. This study demonstrated novel candidate regions in BTA19, 23 and 25 for fatty acid composition.     

Effects of pH and fermentative substrate on ruminal metabolism of fatty acids during short‐term in vitro incubation

The ruminal biohydrogenation of c9,c12‐18:2 can be affected by the fibre/starch ratio of the diet and the ruminal pH. The objectives of this study were to examine independently in vitro the effects of fermentation substrate (hay vs. corn starch) and buffer pH (6 vs. 7) on the biohydrogenation of c9,c12‐18:2 carried out by grape seed oil, focusing on its t11 and t10 pathways, using 6‐h ruminal incubations. The experimental design was a 2 × 2 factorial arrangement. Fermentation substrate and pH affected the C18 fatty acid balance in incubated media, but few interactions were observed. Compared with starch, hay as the fermentation substrate favoured the production of 18:0 (×2.3), all trans‐18:1 isomers (×12.6) and CLA (×6.1), except c9,t11‐CLA, and the disappearance of unsaturated C18 fatty acids, but decreased the production of odd and branched chain fatty acids. Compared with pH 6 buffer, pH 7 buffer resulted in higher c9,c12‐18:2 disappearance and CLA production. For c9,t11‐CLA, an interaction was noticed between the two factors, leading to the highest production in cultures incubated on hay with the 7 pH buffer. Compared with starch, hay as fermentation substrate favoured the activity of t11 producers, which are fibrolytic bacteria, and the production of t10 isomers, possibly due to the presence of potential t10 producers in hay. Low pH resulted in a decreased t11 isomers production and in a slightly increased t10 isomers production, probably due to a modulation of enzymatic or bacterial activity.     

Characterization of bovine neutrophil β2‐adrenergic receptor function

LaBranche, T. P., Ehrich, M. F., Eyre, P. Characterization of bovine neutrophil β₂‐adrenergic receptor function. J. vet. Pharmacol. Therap. doi: 10.1111/j.1365‐2885.2009.01143.x. This study compares bovine leukocyte β‐adrenergic receptor densities to that of the rat, demonstrates for the first time a functional β₂‐adrenergic receptor signaling pathway in steer neutrophils, and investigates the effect of an inflammatory stimulus on that signaling pathway. The β₁‐/β₂‐adrenergic antagonist ^[3H]CGP‐12177 demonstrated that rat lymphocyte specific binding‐site density was highest, followed by steer and dairy cow lymphocytes, and lastly steer and dairy cow neutrophils. The β₂‐adrenergic agonist terbutaline stimulated steer neutrophil adenosine 3,5‐cyclic monophosphate (cAMP) production, an effect increased by inclusion of ≥1 × 10^?8 m phorbol 12‐myristate 13‐acetate (PMA), an activator of protein kinase C. Both terbutaline and the nonselective phosphodiesterase inhibitor 3‐isobutyl‐1‐methylxanthine (IBMX) independently decreased steer neutrophil superoxide anion production in a concentration‐dependent manner, with 1 × 10^?4 m IBMX enhancing both the potency and efficacy of the terbutaline effect (up to 74% reduction in superoxide anion production). Superoxide anion production was also reduced by the synthetic cAMP analog 8‐bromo‐cAMP, which increased the potency of the IBMX effect on superoxide anion production. Taken together, these data demonstrate the presence of a β₂‐adrenergic receptor signaling pathway in bovine neutrophils much like that described in other animal species, as well as the potential for an inflammatory stimulus to alter its function.     

Reducing methane production by supplementation of Terminalia chebula RETZ. containing tannins and saponins

This study investigates the effects of Terminalia chebula Retz. meal supplementation on rumen fermentation and methane (CH₄) production by using an in vitro gas technique. The experimental design was a completely randomized design (CRD) and the dietary treatments were T. chebula supplementation at 0, 4, 8, 12, 16 and 20 mg with 0.5 g of roughage and concentrate ratio at 60:40. The results revealed that cumulative gas production (96 h of incubation) were higher (P < 0.01) with T. chebula supplementation at 12, 16 and 20 mg than other treatments. However, in vitro dry matter degradability (IVDMD) and in vitro organic matter digestibility (IVOMD) were not significantly different among treatments (P > 0.05). The NH₃‐N concentrations tended to quadratically increase with increasing levels of T. chebula in the diet. In addition, total volatile fatty acids (VFA) and propionate concentrations were increased (P < 0.01), while acetate concentration, acetate‐to‐propionate ratio, CH₄ production and protozoal populations were decreased (P < 0.01) when supplemented with T. chebula at 8, 12 and 16 mg, respectively. Based on this study, it could be concluded that supplementation of T. chebula at 12 mg could improve rumen fermentation by reducing CH₄ production and protozoa populations, thus improving in vitro gas production and VFA profiles.     

Effect of α-cyclodextrin-allyl isothiocyanate on ruminal microbial methane production in vitro

The objective of the present study was to investigate the effects of α‐cyclodextrin‐allyl isothiocyanate (CD‐AI) on ruminal microbial methane production and rumen fermentation of corn starch, soluble potato starch or hay plus concentrate (1.5:1) by mixed rumen microorganisms. Diluted rumen fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐AI (0, 0.4, 0.8, 1.6 and 3.2 g/L). The pH of the medium was unchanged by CD‐AI in all substrates. The molar proportion of acetate was decreased and propionate was increased with a corresponding decrease in acetate : propionate ratio (P < 0.05). Total volatile fatty acids and butyrate were increased (P < 0.05). Ammonia‐N was decreased (P < 0.05). Except with soluble potato starch, numbers of protozoa were unchanged after 6 h. As concentration of CD‐AI increased from 0 to 3.2 g/L, fermentation of corn starch, soluble potato starch and hay plus concentrate resulted in decreased (P < 0.05) methane production of 49–100% (6 h) and 14–100% (24 h); 39–100% (6 h) and 16–100% (24 h); and 45–100% (6 h) and 17–100% (24 h), respectively. When hay plus concentrate was used as substrate, methanogenic bacteria were decreased (P < 0.05) with 0.8 g/L of CD‐AI after 6 h. Excluding the lower dose level (0.4 g/L) of CD‐AI, digestibility of neutral detergent fiber of hay plus concentrate was decreased (P < 0.05) after 24 h. A suitable level of CD‐AI could therefore be used as a supplement to inhibit methane production and improve rumen fermentation without detrimental effects on fiber digestion.     

l‐carnitine and fat type in the maternal diet during gestation and lactation modify the fatty acid composition and expression of lipid metabolism‐related genes in piglets

This study examined the influence of adding different amounts of maternal dietary l ‐carnitine and two fat types on fatty acid (FA) composition and the expression of lipid metabolism‐related genes in piglets. The experiment was designed as a 2 × 2 factorial with two fat types (3.5% soyabean oil, SO, and 3.5% fish oil, FO) and two levels of l ‐carnitine (0 and 100 mg/kg) added to the sows' diets. A higher proportion of n‐3 polyunsaturated fatty acids (PUFA) and a lower ratio of n‐6/n‐3 PUFA in sow milk and piglet tissues were observed in the FO groups than in the SO groups. Adding l ‐carnitine increased the proportion of C16:1 in sow milk and decreased n‐3 PUFA in piglet subcutaneous fat. Hepatic peroxisome proliferator‐activated receptor α (PPAR‐α) was more abundantly expressed in piglets from the FO groups than from the SO groups (p < 0.05), whereas stearoyl‐CoA‐desaturase (SCD), sterol regulatory element binding protein‐1 (SREBP1) and ?6‐desaturase (D6D) genes were less expressed in the FO groups compared with piglets from the SO groups. The expression of fatty acid synthase (FAS) genes was decreased in the SO groups with l ‐carnitine compared to that of the other dietary treatments. No differences among dietary treatments were observed with regard to the expression of acetyl‐CoA carboxylase (ACC). In conclusion, FO and l ‐carnitine supplementation in sows affect FA composition and hepatic gene expression in piglets.     

Effect of cyclodextrin diallyl maleate on methane production, ruminal fermentation and microbes in vitro and in vivo

Effects of β‐cyclodextrin diallyl maleate (CD‐M) on methane production, ruminal fermentation and digestibility were studied both in vitro and in vivo. In in vitro study, diluted ruminal fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐M using hay plus concentrate (1.5:1) as a substrate. The CD‐M was added at different concentrations (0, 1.25, 2.5, 5.0 and 7.5 g/L). The pH of the medium and numbers of protozoa were not affected by the addition of CD‐M. Total volatile fatty acids were increased and ammonia‐N was decreased, molar proportion of acetate was decreased and propionate was increased (P < 0.05) by CD‐M. Methane was inhibited (P < 0.05) by 14–76%. The effect of CD‐M on methane production and ruminal fermentation was further investigated in vivo using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay and concentrate mixture (1.5:1) with or without CD‐M (2% of feed dry matter) as a supplement. Ruminal proportion of acetate tended to decrease and that of propionate was increased (P < 0.05) 2 h after CD‐M dosing. Total viable counts, cellulolytic, sulfate reducing, acetogenic bacteria and protozoa were unaffected while methanogenic bacteria were decreased (P < 0.05) by CD‐M. The plasma concentration of glucose was increased, whereas that of urea‐N was decreased (P < 0.05). Methane was inhibited (P < 0.05) from 36.4 to 30.1 L/kg dry matter intake by the addition of CD‐M. Apparent digestibilities of dry matter and neutral detergent fiber were not affected while that of crude protein was increased (P < 0.05) in the medicated steers. These data suggested that dietary supplementation of CD‐M decreased methane production and improved nutrient use.     

Survival of spray‐dried and free‐cells of potential probiotic Lactobacillus plantarum 564 in soft goat cheese

A high viability of probiotics in food product, with a living cells threshold of 10⁷/cfu/g (colony‐forming units/g) is a challenge to achieve in food production. Spray drying is an efficient and economic industrial method for probiotic bacterial preservation and its application in food products. In this study, the survival of free and spray‐dried cells of potential probiotic strain Lactobacillus plantarum 564 after production and during 8 weeks of storage of soft acid coagulated goat cheese was investigated, as well as compositional and sensory quality of cheese. Total bacterial count of spray‐dried Lb. plantarum 564 cells were maintained at the high level of 8.82 log/cfu/g in cheese after 8 weeks of storage, while free‐cell number decreased to 6.9 log/cfu/g. However, the chemical composition, pH values and sensory evaluation between control cheese (C1 sample made with commercial starter culture) and treated cheese samples (C2 and C3, made with the same starter, with the addition of free and spray‐dried Lb. plantarum 564 cells, respectively) did not significantly differ. High viability of potential probiotic bacteria and acceptable sensory properties indicate that spray‐dried Lb. plantarum 564 strain could be successfully used in the production of soft acid coagulated goat cheeses.     

Olive leaves (Olea europea L.) and α‐tocopheryl acetate as feed antioxidants for improving the oxidative stability of α‐linolenic acid‐enriched eggs

Ninety‐six brown Lohmann laying hens were equally assigned into four groups with six replicates. Hens within the control group were fed a corn–soybean‐based diet supplemented with 4% linseed oil. Two other groups were given the same diet further supplemented with 5 or 10 g ground olive leaves/kg feed, while the diet of the fourth group was further supplemented with 200 mg α‐tocopheryl acetate/kg. Supplementing diets with olive leaves had no effect on egg production, feed intake and egg traits. Eggs collected 28 days after feeding the experimental diets were analysed for lipid hydroperoxides and malondialdehyde (MDA) content, fatty acid profile, α‐tocopherol concentrations and susceptibility to iron‐induced lipid oxidation. Olive leaves were also analysed for total and individual phenolics, and total flavonoids, whereas their antioxidant capacity was determined using both the DPPH (1,1‐diphenyl‐2‐picrylhydrazyl) and ABTS (2,2‐azinobis3‐ethylbenzothiazoline‐6‐sulphonic acid) radical scavenging activity assays. Results showed that neither α‐tocopheryl acetate nor olive leaves supplementation exerted (p > 0.05) any effect on the fatty acid composition of n‐3 eggs. Supplementing the diet with 5 g olive leaves/kg had no (p > 0.05) effect on the hydroperoxide levels of n‐3 eggs, while supplementing with 10 g olive leaves/kg or 200 mg α‐tocopheryl acetate/kg, the lipid hydroperoxide levels were reduced (p ≤ 0.05) compared to control. However, although hydroperoxides were reduced, MDA, a secondary lipid oxidation product, was not affected (p > 0.05). Iron‐induced lipid oxidation increased MDA values in eggs from all groups, the increase being higher (p ≤ 0.05) in the control group and the group supplemented with 5 g olive leaves/kg. The group supplemented with 10 g olive leaves/kg presented MDA values lower (p ≤ 0.05) than the control but higher (p ≤ 0.05) than the α‐tocopheryl acetate group, which presented MDA concentrations lower (p ≤ 0.05) than all other experimental diets at all incubation time points.     

Methane production from thermophilic co‐digestion of dairy manure and waste milk obtained from therapeutically treated cows

Methane production from co‐digestion of dairy manure and waste milk, milk from cows treated with antibiotics for mastitis, was tested in a 2 × 4 factorial design. Four different waste milk percentages (w/w): 0% (SM), 10% (SMWM10), 20% (SMWM20) and 30% (SMWM30), were tested with two slurry percentages (w/w): 50% (A) and 25% (B) and the rest being manure at 55°C for 12 days in batch digesters. The results analyzed using a Gompertz model showed SMWM10 produced the highest methane production potential (P_m)/g volatile solids added followed by SM in both A and B. This P_m of SMWM10 in A and B was statistically non‐significant (P > 0.05). More than 96% of cefazolin‐resistant bacteria and 100% of multi‐drug‐resistant bacteria reductions were observed in all the treatments. Inclusion of waste milk at 10% in single stage digester enhances the methane production from dairy manure and could offer added benefit of waste milk treatment and disposal.     

Effect of monensin and vitamin E on milk production and composition of lactating dairy cows

Feeding unsaturated oils to lactating dairy cows impair ruminal biohydrogenation (BH) of unsaturated fatty acids (USFA) and increase ruminal outflow of BH intermediates such as trans‐10, cis‐12 CLA that are considered to be potent inhibitors of milk fat synthesis. Supplementing lactating dairy cow’s rations containing plant origin oils with monensin and/or vitamin E may minimise the formation of trans‐10 isomers in the rumen, thereby preventing milk fat depression. Therefore, this study was conducted to evaluate the effects of monensin and vitamin E supplementation in the diets of lactating dairy cows containing whole cottonseed, as the main source of FA on feed intake, milk production and composition, milk fatty acid profile, efficiency of nitrogen (N) utilisation, efficiency of net energy (NE) utilisation and nutrients digestibilities. Four multiparous Holstein lactating dairy cows (86 ± 41 days in milk) were assigned to a balanced 4 × 4 Latin square design. Each experimental period lasted 21 days with a 14 days of treatment adaptation and a 7 days of data collection. The control diet was a total mixed ration (TMR) consisted of 430 g/kg forage and 570 g/kg of a concentrate mixture on dry matter (DM) basis. Cows were randomly assigned to one of the four dietary treatments including control diet (C), control diet supplemented with 150 mg of vitamin E/kg of DM (E), control diet supplemented with 24 mg of monensin/kg of DM (M) and control diet supplemented with 150 mg of vitamin E and 24 mg of monensin/kg of DM (EM). Dry matter intake (DMI) ranged from 19.1 to 19.5 kg/d and was similar among the dietary treatments. Dietary supplementation with vitamin E or monensin had no effect on milk production, milk fat, protein and lactose concentrations, efficiency of utilisation of nitrogen and net energy for lactation (NE_L). Digestibility of DM, organic matter (OM), crude protein (CP) and ether extract (EE) was not affected by the dietary treatments. Digestibility of neutral detergent fibre (NDF) was higher in cows fed with the M and EM diets in relation to those fed the C and E diets. The concentrations of C4:0, C6:0, C8:0, C10:0, C12:0, C14:0, C15:0, trans‐10‐16:1, cis‐9‐16:1, 17:0, 18:0, trans‐11‐18:1, cis‐9‐18:1, cis‐9, trans‐11 conjugated linoleic acid (CLA), trans‐10, cis‐12 CLA, and 18:3n‐3 FA in milk fat were not affected by the dietary supplementations. While feeding the M diet tended to decrease milk fat concentration of C16:0, the milk fat concentration of C18:2n‐6 FA tended to be increased. Dietary supplementation with vitamin E or monensin had no effect on milk fat concentrations of saturated, unsaturated, monounsaturated, polyunsaturated, short chain and long chain FA, but feeding the M diet numerically decreased milk fat concentration of medium chain fatty acids (MCFA). The results showed that vitamin E and/or monensin supplementations did not improve milk fat content and did not minimise the formation of trans‐10 FA isomers in the rumen when whole cottonseed was included in the diet as the main source of fatty acids.     

添加亚油酸条件下不同剂量硝酸钠对水牛瘤胃体外发酵脂肪酸组成及相关微生物数量的影响

为了探究添加亚油酸条件下不同剂量硝酸钠对水牛瘤胃体外发酵脂肪酸组成及相关微生物数量的影响,本试验选用3头装有永久性瘤胃瘘管的母水牛,以精粗比40∶60为底物进行瘤胃液体外发酵试验。试验组硝酸钠的浓度分别为1、2、3 mg·mL^-1,对照组不加硝酸钠,每组均添加0.25 mg·mL^-1的亚油酸,每组各5个重复。在体外培养3、6、9、12、24 h时测定产气量和甲烷产量,24 h结束培养,测定瘤胃体外发酵参数、脂肪酸含量及瘤胃微生物数量。结果表明：1）添加硝酸钠显著降低了总产气量和甲烷产量（P<0.05）;2）添加硝酸钠pH值和氨态氮（NH₃-N）含量显著升高（P<0.05）,异丁酸和异戊酸含量显著降低（P<0.05）,而对总挥发性脂肪酸（TVFA）含量无显著性影响（P>0.05）;3）添加1 mg·mL^-1硝酸钠,C18：2cis-9,trans-11、C18：2trans-10,cis-12含量和不饱和脂肪酸/饱和脂肪酸（UFA/SFA）显著高于其他组（P<0.05）,且C20：5n3 （EPA）含量显著高于添加3 mg·mL^-1硝酸钠组（P<0.05）,添加2 mg·mL^-1硝酸钠C22：6n3 （DHA）含量显著高于其他组（P<0.05）;4）添加硝酸钠组原虫数量显著低于对照组（P<0.05）,添加2、3 mg·mL^-1硝酸钠产甲烷菌数量显著低于添加1 mg·mL^-1硝酸钠组（P<0.05）,添加1 mg·mL^-1硝酸钠总细菌、真菌、溶纤维丁酸弧菌、蛋白分解丁酸弧菌、非典型丁酸弧菌、亨氏丁酸弧菌数量显著高于其他组（P<0.05）。由此可得,体外法添加1~3 mg·mL^-1硝酸钠和0.25 mg·mL^-1亚油酸在维持水牛TVFA含量不变的情况下显著降低了甲烷含量,且1 mg·mL^-1硝酸钠能促进亚油酸生成共轭亚油酸（CLA）,优化脂肪酸组成,并能增加大多数瘤胃微生物的数量。     

Trans10, cis12 conjugated linoleic acid increases triacylglycerol accumulation in goat mammary epithelial cells in vitro

Trans10, cis12 conjugated linoleic acid (t10c12‐CLA) is well‐established in decreasing milk fat content and causing milk fat depression (MFD) in dairy cattle and goats. However, the detailed mechanisms of its effect are not completely understood. Therefore, we used goat mammary epithelial cells (GMECs) to further study the molecular mechanisms whereby t10c12‐CLA regulates milk fat synthesis. The optimal concentration of t10c12‐CLA (100 μmol/L) for cell culture was determined through a cell vitality and morphology assay, and evaluation of abundance of apoptosis‐related proteins. Oil red O stain indicated that t10c12‐CLA increased concentration of cytoplasmic lipid droplets. Furthermore, t10c12‐CLA increased the intracellular triacylglycerol (TG) content (P < 0.05). Among 16 genes related to lipid metabolism that were measured by quantitative real‐time PCR, t10c12‐CLA down‐regulated (P < 0.05) genes involved in de novo fatty acid synthesis including FASN, ACACA and SCD1, and also down‐regulated the protein expression of FASN and SCD1 but up‐regulated (P < 0.05) the expression of CD36 and ADRP. Overall, the data indicate that a side effect of de novo fatty acid synthesis inhibition by t10c12‐CLA is the up‐regulation of fatty acid uptake and accumulation of lipid droplets in GMECs. The biologic reason for such an effect merits further study.     

Genome‐wide association studies reveal additional related loci for fatty acid composition in a Duroc pig multigenerational population

The aim of the present study was to detect quantitative trait loci affecting fatty acid composition in back fat and intramuscular fat in a Duroc pig population comprising seventh‐generation pedigrees using genome‐wide association studies (GWAS). In total, 305 animals were genotyped using single nucleotide polymorphisms (SNPs) array and five selected SNPs from regions containing known candidate genes related to fatty acid synthesis or metabolism. In total, 24 genome‐wide significant SNP regions were detected in 12 traits, and 76 genome‐wide suggestive SNP regions were detected in 33 traits. The Sus scrofa chromosome (SSC) 7 at 10.3 Mb was significantly associated with C17:0 in intramuscular fat, while the SSC9 at 13.6 Mb was significantly associated with C14:0 in intramuscular fat. The SSC12 at 1.0 Mb was significantly associated with C14:0 in back fat and the SSC14 at 121.0 Mb was significantly associated with C18:0 in intramuscular fat. These regions not only replicated previously reported loci containing some candidate genes involved in fatty acid composition (fatty acid synthase and stearoyl‐CoA desaturase) but also included several additional related loci.     

Effects of calcium soap of rice bran oil fatty acids supplementation alone and with DL‐α‐tocopherol acetate in lamb diets on performance,digestibility, ruminal parameters and meat quality

Thirty‐six Malpura lambs (28 day old and 6.7 ± 0.25 kg BW) were distributed equally in three groups having six males and six female. They were ad libitum fed individually three different experimental diets containing calcium soap of fatty acids (CA‐FA) at 0 (T₁) and 40 (T₂ and T₃) g/kg concentrate up to six months of age. Animals in T₃ were supplemented additionally with 40 mg DL‐α‐tocopherol acetate/kg of concentrate. The roughage moiety included ad libitum dry Prosopis cineraria and fresh Azadirachata indica leaves. All the lambs were allowed to suckle from their dam up to weaning (90 day of age). Supplementation of Ca‐FA improved weight gain and feed conversion ratio during both pre‐ (28–90 days) and post‐weaning (91–180 days) phases; however, no effect of DL‐α‐tocopherol was observed. Metabolic parameters during post‐weaning phase revealed non‐significant effect on digestibility but improved nitrogen balance in the test groups. The effect on biochemical attributes did not show any significant alteration in ruminal parameters, blood biochemicals and urinary purine derivatives. Carcass traits revealed higher (p < 0.05) dressing yield and loin eye area with Ca‐FA supplementation. The value of thiobarbituric reactive substances for nuggets prepared from frozen carcasses revealed significant (p < 0.05) reduction in T₃ compared to the other dietary groups. Fatty acid profile of adipose tissue revealed higher (p < 0.001) 9‐octadecanoic, 9‐12‐octadecadienoic, polyunsaturated fatty acids (PUFA), higher ratio of PUFA/saturated fatty acids (SFA), ω‐6/ω‐3 and lower SFA in Ca‐FA‐supplemented groups. It is concluded that supplementation of 40 g/kg calcium soap prepared from industrial grade rice bran oil in lamb ration provided additional energy intake, improved N utilization, gain and feed conversion ratio besides improving dressing yield and meat quality with CLA enriched fatty acid profile. DL‐α‐tocopherol acetate when supplemented at 40 mg/kg feed reduced lipid oxidation of meat products thus improving its keeping quality.     

Effects of Bacillus subtilis B10 spores on viability and biological functions of murine macrophages

This study was conducted to investigate the effects of Bacillus subtilis B10 spores on the viability and biological functions of murine macrophage. RAW 264.7 cells were stimulated both with and without B. subtilis B10 spores for 12 h. Then cell viability was determined to evaluate the cytotoxic effect of B. subtilis B10 spores to the cells, and the activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH), the production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS), and the secretion of inflammatory cytokines were measured to analyze the functions of macrophages. The results showed that B. subtilis B10 spores were not harmful to RAW 264.7 cells and they also strongly enhanced the activities of ACP and LDH (P < 0.01), remarkably increased NO and iNOS production (P < 0.01), and significantly stimulated the secretion of pro‐inflammatory cytokines, including tumor necrosis factor‐alpha (TNF‐α), interferon‐gamma (IFN‐γ), interleukin‐1 beta (IL‐1β), IL‐6, IL‐8 and IL‐12 (P < 0.01) while they reduced anti‐inflammatory cytokine IL‐10 (P < 0.01). The outcomes suggest that B. subtilis B10 spores are not only safe for murine macrophages, but also can activate these cells and enhance their immune function. The above findings suggest that B. subtilis B10 spores are potentially probiotic.     

Medium‐chain glycerides affect gut morphology,immune‐ and goblet cells in post‐weaning piglets: In vitro fatty acid screening with Escherichia coli and in vivo consolidation with LPS challenge

The influence of medium‐chain glycerides on performance and gastrointestinal well‐being in weaning piglets was assessed. First, caproic (C6), caprylic (C8) and capric (C10) acid activity against Escherichia coli was screened in vitro. Pig flora of the whole small intestine was used as inoculum. Seven in vitro incubations were done in duplicate at pH = 3 and 5: C10 (15 mM), C8 (12 mM), C6 (15, 12, 10 mM), a non‐incubated‐negative control and incubated negative control. Culture suspensions were plated on E. coli‐selective agar. Controls showed bacterial growth. C6 and C8 showed no growth at both pH‐values, where C10 showed growth at pH = 5. Secondly, an in vivo study was done with 80 weaned piglets over 42 days, housed in pens of eight animals (five pens/treatment), fed a basal diet containing broken rice/soya bean meal/fish meal and supplemented with C6 and C8 in medium‐chain glyceride form (MCT6/8, 0.175%) or antibiotic growth promoter (AGP, 0.020%) (Kasetsart University, Thailand) serving as control. Feed intake, daily gain and feed‐to‐gain ratio did not differ between MCT6/8 and AGP. Per replicate, two random selected piglets were challenged intravenously with E. coli‐lipopolysaccharide (LPS) or saline solution (S) at Days 21 and 28. All challenged animals were sacrificed; blood and digestive tract samples (jejunum/ileum) were collected at Day 35. LPS challenge consistently reduced villus height and crypt depth for MCT6/8 and AGP. However, LPS‐challenged piglets supplemented with MCT6/8 restored villus height, where AGP did not. MCT6/8 piglets had higher serum IgA, more jejunal IgA‐positive plasma cells and goblet cells than AGP. At the ileal level, results were similar, though less pronounced. The present study offers new insight in the benefits of MCT6/8 over AGP in the post‐weaning period. There is in vitro anti‐microbial action of C6 and C8 on E. coli. In vivo, MCT6/8 also has protective effects in the small intestine that may result in growth promotion.     

Papaya (Carica papaya) leaf methanolic extract modulates in vitro rumen methanogenesis and rumen biohydrogenation

Papaya leaf methanolic extract (PLE) at concentrations of 0 (CON), 5 (LLE), 10 (MLE) and 15 (HLE) mg/250 mg dry matter (DM) with 30 mL buffered rumen fluid were incubated for 24 h to identify its effect on in vitro ruminal methanogenesis and ruminal biohydrogenation (BH). Total gas production was not affected (P > 0.05) by addition of PLE compared to the CON at 24 h of incubation. Methane (CH₄) production (mL/250 mg DM) decreased (P < 0.05) with increasing levels of PLE. Acetate to propionate ratio was lower (P <0.05) in MLE (2.02) and HLE (1.93) compared to the CON (2.28). Supplementation of the diet with PLE significantly (P <0.05) decreased the rate of BH of C18:1n‐9 (oleic acid; OA), C18:2n‐6 (linoleic acid; LA), C18:3n‐3 (linolenic acid; LNA) and C18 polyunsaturated fatty acids (PUFA) compared to CON after 24 h incubation, which resulted in higher concentrations of BH intermediates such as C18:1 t11 (vaccenic acid; VA), c9t11 conjugated LA (CLA) (rumenic acid; RA) and t10c12 CLA. Real‐time PCR analysis indicated that the total bacteria, total protozoa, Butyrivibrio fibrisolvens and methanogen population in HLE decreased (P <0.05) compared to CON, but the total bacteria and B. fibrisolvens population were higher (P < 0.05) in CON compared to the PLE treatment groups.