Effects of Rubus coreanus byproducts on intestinal microbiota and the immune modulation.

OBJECTIVE: Although the efficacy of Rubus coreanus (RC) byproducts as a feed additive has been recognized, its effects on intestinal microorganisms and the immune system are still unknown. METHODS: Six-week-old male rats were treated with 0.5% RC (T1), 1.0% RC (T2), and 1.5% RC (T3) for 4 weeks. RESULTS: We found that treatment with RC byproducts significantly increased the daily gain of body weight and feed intake. Treg-cell differentiation was enhanced in the mesenteric lymph nodes and spleen from the rats fed with RC byproducts. Illumina sequencing showed that bacteria in the phylum Firmicutes decreased and while those in the phylum Bacteroidetes increased in RC-treated groups. Particularly, the pathogenic microorganisms in the family Peptococcaceae decreased, and the non-pathogenic families Lachnospiraceae and S24-7 increased. Quantitative polymerase chain reaction analysis showed that the RC byproducts increased the lactic acid bacteria Bifidobacterium spp., Oscillospira spp., Leuconostoc citreum, and Weissella cibaria in a concentration-dependent manner. CONCLUSION: RC byproducts may be effective in immunomodulation by affecting intestinal microorganisms.

Cloning and Characterization of an Endoglucanase Gene from Actinomyces sp. Korean Native Goat 40.

A gene from Actinomyces sp. Korean native goat (KNG) 40 that encodes an endo-ß-1,4-glucanase, EG1, was cloned and expressed in Escherichia coli (E. coli) DH5α. Recombinant plasmid DNA from a positive clone with a 3.2 kb insert hydrolyzing carboxyl methyl-cellulose (CMC) was designated as pDS3. The entire nucleotide sequence was determined, and an open-reading frame (ORF) was deduced. The ORF encodes a polypeptide of 684 amino acids. The recombinant EG1 produced in E. coli DH5α harboring pDS3 was purified in one step using affinity chromatography on crystalline cellulose and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/zymogram analysis of the purified enzyme revealed two protein bands of 57.1 and 54.1 kDa. The amino terminal sequences of these two bands matched those of the deduced ones, starting from residue 166 and 208, respectively. Putative signal sequences, a Shine-Dalgarno-type ribosomal binding site, and promoter sequences related to the consensus sequences were deduced. EG1 has a typical tripartite structure of cellulase, a catalytic domain, a serine-rich linker region, and a cellulose-binding domain. The optimal temperature for the activity of the purified enzyme was 55°C, but it retained over 90% of maximum activity in a broad temperature range (40°C to 60°C). The optimal pH for the enzyme activity was 6.0. Kinetic parameters, Km and Vmax of rEG1 were 0.39% CMC and 143 U/mg, respectively.

Physiological properties of Scomber japonicus meat hydrolysate prepared by subcritical water hydrolysis.

The health-beneficial biological activities, including antioxidant and tyrosinase inhibitory activities, of Scomber japonicus muscle protein hydrolysates prepared by subcritical water hydrolysis were investigated. After 5 min of subcritical hydrolysis at 140 degrees C, 59.76% of S. japonicus muscle protein was hydrolyzed, the highest degree of hydrolysis in all the groups were tested. According to the response surface methodology results, as the reaction temperature and reaction time became lower and shorter, the yield became higher. The highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity (90.63%) occurred in hydrolysates treated at 140 degrees C for 5 min, and the highest tyrosinase inhibitory activity (65.54%) was identified in hydrolysates treated at 200 degreesC for 15 min. Changes in the molecular weight distribution of S. japonicus muscle proteins after subcritical water hydrolysis were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Subcritical water hydrolysis is a suitable technique for obtaining S.japonicus muscle protein hydrolysates with useful biological activities, within a short time (5-15 min).

Effect of Soybean Meal and Soluble Starch on Biogenic Amine Production and Microbial Diversity Using In vitro Rumen Fermentation.

This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in NH3-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest NH3-N concentration among all groups after 24 h of incubation. Increasing methane (CH4) concentrations were observed with time, and CH4 concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, NH3-N, CH4, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

Inhibitory effects of seaweed extracts on the growth of the vaginal bacterium Gardnerella vaginalis.

Of 44 species of seaweed screened for potential anti-Gardnerella vaginalis activity, 27 (61.4%) showed antimicrobial activity by the agar disk-diffusion method. Among them, the strongest activities against the pathogen were exhibited by Chlorophyta, with Ulva pertusa producing an 11.3-mm zone of inhibition at 5 mg disk⁻¹. The MIC values of U. pertusa extracts against both G. vaginalis KCTC 5096 and KCTC 5097, the main cause of vaginosis, were 312 µg ml⁻¹, while the MIC values against both Candida albicans KCTC 7270 and KCTC 7965, the main cause of candidiasis, were 2.5 mg ml⁻¹. Against Lactobacillus gasseri KCTC 3173 and Lactobacillus jensenii KCTC 5194, members of the normal vaginal microflora, no inhibitory effect was seen even at 10 mg ml⁻¹. To identify the primary active compounds, a U. pertusa powder was successively fractionated according to polarity, and the main active agents against G. vaginalis KCTC 5096 were determined to be nitrogenous compounds (156 µg ml⁻¹ of the MIC value). According to these results, it was suggested that extracts of the seaweed U. pertusa are valuable for the development of natural therapeutic agents for treating women with bacterial vaginosis.

Seasonal variation of antibacterial activities in the green alga Ulva pertusa Kjellman.

The present study was performed to screen out the extracts of algae and assess the seasonal variation in antimicrobial activity of Ulva pertusa against Gardnerella vaginalis. Seasonal variation in antibacterial activity was observed, with the extracts showing no activity during summer and autumn, and showing antibacterial activity from early winter (December) to middle spring (April). The maximum value of antimicrobial activity (6.5 mm inhibition zone at 5 mg disk(-1)) of U. pertusa against G. vaginalis was observed in April. Otherwise, for both chlorophyll a and b, the highest content (2.87 mg g(-1) and 1.37 mg g(-1)) was observed in March 2009. These results may reflect variation in cellular chemical compositions such as secondary metabolite(s) rather than chlorophyll and biological activities according to season.

Efficacious rat model displays non-toxic effect with Korean beechwood creosote: a possible antibiotic substitute.

Wood creosote, an herbal anti-diarrheal and a mixture of major volatile compounds, was tested for its non-toxicological effects, using a rat model, with the objective to use the creosote as an antibiotic substitute. A total of 30 Sprague-Dawley rats were studied to form five groups with 6 rats each. Korea beechwood creosote was supplemented into three test groups with 0.03 g/kg, 0.07 g/kg and 0.1 g/kg body weight/day without antibiotic support, along with a positive control of Apramycin sulphate (at 0.5% of the daily feed) and a negative control. Korean beechwood creosote supplementation showed no negative effect on the body weight gain in comparison to the negative and the positive control groups and the feed conversion ratio was also comparable with that of the control groups. The clinical pathology parameters studied were also under the umbrella of normal range, including liver specific enzymes, blood glucose, total protein, blood urea nitrogen (BUN), which indicated no toxic effect of creosote at the given doses. The non-hepatotoxic effect was also confirmed using hepatic damage specific molecular markers like Tim-p1, Tim-p2 and Tgf-ß1. The results suggested that Korean beechwood may be used as antibiotic substitute in weanling pigs feed without any toxic effect on the body. Although the antimicrobial properties of creosote were not absolutely similar to those of apramycin sulphate, they were comparable.

Swine liver methylomes of Berkshire, Duroc and Landrace breeds by MeDIPS.

Using a methyl-DNA immunoprecipitation technique in combination with next-generation deep sequencing, we conducted comprehensive DNA methylation profiling of liver genomes from three pig breeds: Berkshire, Duroc and Landrace. The profiles revealed that the distribution patterns of methylation signals along the genome are conserved among the three pig breeds. Specifically, many signals in coding genes were found in introns, and most signals in the repetitive elements were identified in non-long terminal repeat (LTR) retrotransposons such as long and short interspersed repetitive elements, implying a significant association with alternative splicing and expression of retrotransposable elements respectively. Differentially methylated regions among the three pig breeds were identified in the non-LTR retrotransposons, suggesting that they may lead to differential retrotransposable element activity. Altogether, this study provides advanced swine methylome data and valuable resources for understanding the function of DNA methylation in the evolutionary divergence of different pig breeds.

Influence of penicillin on microbial diversity of the cecal microbiota in broiler chickens.

Antibiotic growth promoters have been used for growth promotion of chickens in poultry industry since 1940. Recently, concerns have been raised to the use of antibiotic growth promoters in livestock due to development of antibiotic resistance in bacteria. The objective of our study was to investigate the effect of penicillin supplementation in the feed on cecal microbiota of broiler chickens. Two groups (n = 30) of chickens were fed corn-soybean meal diets with and without supplementation of penicillin at the concentration of 55 mg/kg (ANT vs. CON, respectively). At 18 d of age, the ANT group had significantly (P ≤ 0.05) higher mean BW than the CON group (668.6 vs. 570.0 g). Cecal samples of 5 randomly selected birds were pooled from each group and used for genomic DNA isolation and PCR amplification of 16S rRNA gene; 454 pyrosequencing of the amplicons resulted in 7,881 and 11,214 sequence reads for ANT and CON groups, respectively. Phylogenetic analysis indicated that penicillin supplementation in the ANT group resulted in an elevated proportion of phylum Firmicutes from 58.15 to 91.5% and a decreased proportion of phylum Bacteroidetes from 31.1 to 2.96% compared with the CON group. Recent studies conducted in humans, pigs, and mice have shown a similar shift in gut microbiota in obese individuals compared with the lean ones, indicating that this microbial shift could be responsible for the increase in energy harvest and BW. The results of this study suggest that the growth-promoting effect of penicillin supplementation in broilers may be mediated by a similar microbial process.

Effects of seaweed Laminaria japonica extracts on skin moisturizing activity in vivo.

Twelve species of edible seaweed from the coast of Korea were screened for skin moisturizing activity. We placed the lead of a Corneometer on an approximately 6-cm2 test area of the forearm and measured both untreated skin (control) and skin treated with test moisturizing creams either containing or not containing 5% water:propylene glycol (50:50) extracts of seaweeds. Over the 8-h observation period, the strongest activity of the Laminaria japonica extracts occurred at the 2-h period. For the 10% extract, hydration with the L. japonica extract increased by 14.44% compared with a placebo. Transepidermal water loss (TEWL) was also measured using a test cream with 10% L. japonica extract. For up to 8 h after applying the creams, TEWL was decreased to 4.01 g/cm2, which was approximately 20% of that seen with the control. We suggest that the L. japonica extract hydrates skin via the humectants and hydrocolloids that it contains. To confirm the safety of L. japonica extracts, we performed a patch test on human skin. The results suggested that at moderate doses humans can safely use the extracts. For commercial applications, we evaluated the physicochemical characteristics of the test cream products, including Hunter L, a, and b values; pH; refractive index; and coefficient of viscosity. L. japonica extract did not affect overall formulations of the test cream product in any of the tested aspects. These results suggest that L. japonica extract is a promising ingredient in moisturizing formulations.

The antibacterial activity of various saturated and unsaturated fatty acids against several oral pathogens.

The antibacterial activity of various saturated fatty acids (SFA) and unsaturated fatty acids (USFA) against different oral pathogens which are implicated in the cause of dental caries, stomatitis, gingivitis, and periodontitis was examined. The saturated fatty acids Pa, StA and ArA, and the unsaturated omega-7 fatty acids PLA and omega-9 fatty acids OA showed either none to low antimicrobial activity against all of the 12 oral pathogenic strains used in this study. In contrast, the omega-3 PUFAs, ALA, SDA, EPA and DHA, and the omega-6 PUFAs, LA, GLA, and AA showed considerable antimicrobial activity against 8, 7, 6 and 5 strains, and 6, 10 and 5 strains, respectively. In particular, the omega-3 and omega-6 PUFAs showed strong antimicrobial activity against Porphyromonas gingivalis KCTC 381, the cause of periodontitis, and against Aggregatibacter segnis KCTC 5968, Fusobacterium nucleatum subsp. Polymorphum KCTC 5172 and Prevotella intermedia KCTC 25611, all organisms implicated in the cause of gingivitis. To date, no bacterial resistance to free fatty acids has been encountered and no resistance phenotype has emerged. Therefore, these results suggest that PUFAs may be useful in the development of therapeutic agents for oral diseases, and in particular, in the development of agents that have minimal side effects and against which there is no bacterial resistance.

Effects of lactic acid bacteria fermented feed and three types of lactic acid bacteria (L. plantarum, L. acidophilus, B. animalis) on intestinal microbiota and T cell polarization (Th1, Th2, Th17, Treg) in the intestinal lymph nodes and spleens of rats.

OBJECTIVE: In this study, we investigated the effects of Rubus coreanus-derived lactic acid bacteria (LAB) fermented feed (RC-LAB fermented feed) and three types of LAB (Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium animalis) on the expression of transcription factors and cytokines in Th1, Th2, Th17, and Treg cells in the intestinal lymph nodes and spleens of rats. In addition, the effect on intestinal microbiota composition and body weight was investigated. METHODS: Five-week-old male rats were assigned to five treatments and eight replicates. The expression of transcription factors and cytokines of Th1, Th2, Th17, and Treg cells in the intestinal lymph nodes and spleens was analyzed using real-time reverse transcriptase polymerase chain reaction assays. Intestinal tract microbiota compositions were analyzed by next-generation sequencing and quantitative polymerase chain reaction assays. RESULTS: RC-LAB fermented feed and three types of LAB increased the expression of transcription factors and cytokines in Th1, Treg cells and Galectin-9, but decreased in Th2 and Th17 cells. In addition, the intestinal microbiota composition changed, the body weight and Firmicutes to Bacteroidetes (F/B) ratio decreased, and the relative abundance of LAB increased. CONCLUSION: LAB fermented feed and three types of LAB showed an immune modulation effect by inducing T cell polarization and increased LAB in the intestinal microbiota.

Hemato-biochemical and Cortisol Profile of Holstein Growing-calves Supplemented with Vitamin C during Summer Season.

Effect of vitamin C (VC) on biochemical, hematological and cortisol profile of growing Holstein calves during summer was investigated. Eighteen calves between 14 and 16 weeks of age were divided equally into two groups. One group received a diet supplemented with VC (20 g/d) for 60 days, while other non-supplemented diet fed group served as a control (CON). The temperature humidity index (THI) was recorded and computed during the experiment. From days 0 to 60, the THI exceeded 70. Blood samples were collected from the jugular vein of each calf at days 0, 15, 30, 45 and 60. Serum albumin and total protein decreased (p<0.05) in CON and VC calves with age. Serum glutamic-oxaloacetic transaminase concentrations were not affected by treatments. Serum creatinine, albumin and glutamic-pyruvic transaminase concentrations were higher in calves in the VC group than the CON group. While red blood cells, hemoglobin and hematocrit were lower (p<0.05) in VC calves, mean corpuscular volume, mean corpuscular hemoglobin, red blood cell distribution width and mean platelet volume were higher (p<0.05) in these VC supplemented calves. Leukocyte parameters including white blood cells and full term for lymphocytes were not affected by the treatments. Also, serum cortisol was not affected by treatments. At day 15, 30 and 45, the total VC in plasma was higher (p<0.05) in calves fed with VC. In conclusion, serum cortisols were not affected by plasma VC concentration, while some blood parameters were positively influenced in calves fed with VC.

Inhibition of oral pathogens and collagenase activity by seaweed extracts.

Fifty-seven species of common seaweed from the Coast of Korea were screened for antimicrobial (i.e. inhibition of Prevotella intermedia and Porphyromonas gingivalis growth) activity. As a source of bioactive compounds, seaweeds can produce many secondary metabolites with a variety of activities. Using the agar diffusion method, only 17 species (29.8%) showed inhibitory activity. Of these, methanol extracts of Enteromorpha linza, Sargassum sagamianum, and Ulva pertusa showed strong inhibitory effects against both P. intermedia and P. gingivalis. The MIC values of E. linza, S. sagamianum, and U. pertusa extracts against P. intermedia were 625, 78 and 625 microg ml(-1) and those against P. gingivalis were 312, 156 and 625 microg ml(-1), respectively. When these three species' extracts were separated into five fractions according to their polarity, the main active agents were determined to be phenolic compounds. We then compared the antimicrobial activities of these phenolic compounds against various periodontal pathogens using a MIC test. Phenolic compound containing extracts at concentrations of 10 to 100 microg ml(-1) showed a moderate to significant inhibitory effect on collagenase 1,2 and 3 activity.

Intestinal microbial composition changes induced by Lactobacillus plantarum GBL 16, 17 fermented feed and intestinal immune homeostasis regulation in pigs.

In this study, Rubus coreanus (R. coreanus) byproducts with high polyphenol content were fermented with R. coreanus-derived lactic acid bacteria (Lactobacillus plantarum GBL 16 and 17). Then the effect of R. coreanus-derived lactic acid bacteria fermented feed (RC-LAB fermented feed) with probiotics (Bacillus subtills, Aspergillus oryzae, Yeast) as a feed additive for pigs on the composition of intestinal microbes and the regulation of intestinal immune homeostasis was investigated. Seventy-two finishing Berkshire pigs were randomly allotted to four different treatment groups and 18 replicates. RC-LAB fermented feed with probiotics increased the genera Lactobacillus, Streptococcus, Mitsuokella, Prevotella, Bacteroides spp., Roseburia spp., and Faecalibacterium prausnitzii, which are beneficial bacteria of the digestive tract of pigs. Also, RC-LAB fermented feed with probiotics decreased the genera Clostridium, Terrisporobacter, Romboutsia, Kandleria, Megasphaera and Escherichia, which are harmful bacteria. In particular, the relative abundance of the genera Lactobacillus and Streptococcus increased by an average of 8.51% and 4.68% in the treatment groups and the classes Clostridia and genera Escherichia decreased by an average of 27.05% and 2.85% in the treatment groups. In mesenteric lymph nodes (MLN) and spleens, the mRNA expression of transcription factors and cytokines in Th1 and Treg cells increased and the mRNA expression of Th2 and Th17 transcription factors and cytokines decreased, indicating a regulatory effect on intestinal immune homeostasis. RC-LAB fermented feed regulates gut immune homeostasis by influencing the composition of beneficial and detrimental microorganisms in the gut and regulating the balance of Th1/Th2 and Th17/Treg cells.

Increasing buffering capacity enhances rumen fermentation characteristics and alters rumen microbiota composition of high-concentrate fed Hanwoo steers.

The buffering capacity of buffer agents and their effects on in vitro and in vivo rumen fermentation characteristics, and bacterial composition of a high-concentrate fed Hanwoo steers were investigated in this study. Treatments were comprised of CON (no buffer added), BC0.3% (low buffering capacity, 0.3% buffer), BC0.5% (medium buffering capacity, 0.5% buffer), and BC0.9% (high buffering capacity, 0.9% buffer). Four Hanwoo steers in a 4 × 4 Latin square design were used for the in vivo trial to assess the effect of treatments. Results on in vitro experiment showed that buffering capacity, pH, and ammonia-nitrogen concentration (NH3-N) were significantly higher in BC0.9% and BC0.5% than the other treatments after 24 h incubation. Individual and total volatile fatty acids (VFA) concentration of CON were lowest compared to treatment groups. Meanwhile, in vivo experiment revealed that Bacteroidetes were dominant for all treatments followed by Firmicutes and Proteobacteria. The abundances of Barnesiella intestinihominis, Treponema porcinum, and Vibrio marisflavi were relatively highest under BC0.9%, Ruminoccocus bromii and Succiniclasticum ruminis under BC0.5%, and Bacteroides massiliensis under BC0.3%. The normalized data of relative abundance of observed OTUs' representative families have grouped the CON with BC0.3% in the same cluster, whereas BC0.5% and BC0.9% were clustered separately which indicates the effect of varying buffering capacity of buffer agents. Principal coordinate analysis (PCoA) on unweighted UniFrac distances revealed close similarity of bacterial community structures within and between treatments and control, in which BC0.9% and BC0.3% groups showed dispersed community distribution. Overall, increasing the buffering capacity by supplementation of BC0.5% and and BC0.9% buffer agents enhanced rumen fermentation characteristics and altered the rumen bacterial community, which could help prevent ruminal acidosis during a high-concentrate diet.

Seasonal Influence on Rumen Microbiota, Rumen Fermentation, and Enteric Methane Emissions of Holstein and Jersey Steers under the Same Total Mixed Ration.

Seasonal effects on rumen microbiome and enteric methane (CH4) emissions are poorly documented. In this study, 6 Holstein and 6 Jersey steers were fed the same total mixed ration diet during winter, spring, and summer seasons under a 2 × 3 factorial arrangement for 30 days per season. The dry matter intake (DMI), rumen fermentation characteristics, enteric CH4 emissions and rumen microbiota were analyzed. Holstein had higher total DMI than Jersey steers regardless of season. However, Holstein steers had the lowest metabolic DMI during summer, while Jersey steers had the lowest total DMI during winter. Jersey steers had higher CH4 yields and intensities than Holstein steers regardless of season. The pH was decreased, while ammonia nitrogen concentration was increased in summer regardless of breed. Total volatile fatty acids concentration and propionate proportions were the highest in winter, while acetate and butyrate proportion were the highest in spring and in summer, respectively, regardless of breed. Moreover, Holstein steers produced a higher proportion of propionate, while Jersey steers produced a higher proportion of butyrate regardless of season. Metataxonomic analysis of rumen microbiota showed that operational taxonomic units and Chao 1 estimates were lower and highly unstable during summer, while winter had the lowest Shannon diversity. Beta diversity analysis suggested that the overall rumen microbiota was shifted according to seasonal changes in both breeds. In winter, the rumen microbiota was dominated by Carnobacterium jeotgali and Ruminococcus bromii, while in summer, Paludibacter propionicigenes was predominant. In Jersey steers, Capnocytophaga cynodegmi, Barnesiella viscericola and Flintibacter butyricus were predominant, whereas in Holstein steers, Succinivibrio dextrinosolvens and Gilliamella bombicola were predominant. Overall results suggest that seasonal changes alter rumen microbiota and fermentation characteristics of both breeds; however, CH4 emissions from steers were significantly influenced by breeds, not by seasons.

Galectin-9 Induced by Dietary Prebiotics Regulates Immunomodulation to Reduce Atopic Dermatitis Symptoms in 1-Chloro-2,4-Dinitrobenzene (DNCB)-Treated NC/Nga Mice.

Atopic dermatitis (AD) is a skin disorder that causes chronic itch. We investigated the inhibitory effects of a mixture of prebiotic short-chain galacto-oligosaccharides and long-chain fructooligosaccharides (scGOS/lcFOS), inulin, or ß-glucan on AD development in 1-chloro-2,4- dinitrobenzene (DNCB)-treated NC/Nga mice. Mice were randomly assigned to six groups: untreated mice, AD control, positive control (DNCB-treated NC/Nga mice fed a dietary supplement of Zyrtec), and DNCB-treated NC/Nga mice fed a dietary supplement of prebiotics such as scGOS/lcFOS (T1), inulin (T2), or ß-glucan (T3). The prebiotic treatment groups (T1, T2, and T3) showed suppression of AD symptoms, Th2 cell differentiation, and AD-like skin lesions induced by DNCB. In addition, prebiotic treatment also reduced the number of microorganisms such as Firmicutes, which is associated with AD symptoms, and increased the levels of Bacteroidetes and Ruminococcaceae, which are associated with alleviation of AD symptoms. Our findings demonstrate the inhibitory effects of prebiotics on AD development by improving the Th1/Th2 cytokine balance and beneficial symbiotic microorganisms in in vitro and in vivo models.

Comparison of gene expression profiling between bovine subcutaneous and intramuscular adipose tissues by serial analysis of gene expression.

The deposition of intramuscular adipose fat is an important factor in the quality of beef, particularly in Korean cattle and Japanese cattle. A better understanding of the differential mechanisms regulating the deposition and release of fat between intramuscular and external adipose tissues, particularly subcutaneous adipose tissues, would prove vital in understanding the deposition of intramuscular fat. A SAGE (serial analysis of gene expression) method was used to identify functionally active genes in adipose tissues and to compare the global gene expression profile between the subcutaneous and the intramuscular adipose tissues in Korean cattle. We detected 11268 tags, representing 3761 transcripts in the SAGE library of the subcutaneous adipose tissues. We detected 10830 tags, representing 2918 transcripts in the intramuscular adipose tissues. Comparison of gene expression patterns between subcutaneous adipose tissues and intramuscular adipose tissues revealed that 82 genes were differentially expressed at >2-fold levels. Among them, expression levels of 50 genes were higher in subcutaneous adipose tissues compared with intramuscular tissues, whereas those of 32 genes were higher in intramuscular adipose tissues. The genes that displayed >2-fold higher expression in subcutaneous adipose tissues included secreted protein acidic and rich in cysteine, annexin II, thyroid hormone receptor interactor 1, ferritin heavy chain polypeptide 1, cofilin, diacylglycerol O-acyltransferase and thymosin beta 4. Those that showed >2-fold higher expression levels in intramuscular adipose tissues included troponin t3, heat shock 70 kDa protein 1 and NMT2 (N-myristoyltransferase 2). Expression patterns of NMT2 and troponin t3 genes were examined during differentiation of 3T3-LI preadipocytes. NMT2 gene was actively expressed at both the growing stage of preadipocytes and the initial stage of differentiation of 3T3-L1 adipocytes, but its expression was reduced at the active stage of adipocyte differentiation. Troponin t3 gene was not expressed at the growing stage of preadipocyte, but it was actively expressed during differentiation of adipocytes. This implies that troponin t3 may play a positive role in intramuscular fat deposition of beef. Our SAGE experiment provides a differential gene expression profile in subcutaneous and intramuscular adipose tissues of Korean cattle. This information will be useful in developing method(s) to produce high-marble beef.

Rumen fermentation and microbial community composition influenced by live Enterococcus faecium supplementation.

Supplementation of appropriate probiotics can improve the health and productivity of ruminants while mitigating environmental methane production. Hence, this study was conducted to determine the effects of Enterococcus faecium SROD on in vitro rumen fermentation, methane concentration, and microbial population structure. Ruminal samples were collected from ruminally cannulated Holstein-Friesian cattle, and 40:60 rice straw to concentrate ratio was used as substrate. Fresh culture of E. faecium SROD at different inclusion rates (0, 0.1%, 0.5%, and 1.0%) were investigated using in vitro rumen fermentation system. Addition of E. faecium SROD had a significant effect on total gas production with the greatest effect observed with 0.1% supplementation; however, there was no significant influence on pH. Supplementation of 0.1% E. faecium SROD resulted in the highest propionate (P = 0.005) but the lowest methane concentration (P = 0.001). In addition, acetate, butyrate, and total VFA concentrations in treatments were comparatively higher than control. Bioinformatics analysis revealed the predominance of the bacterial phyla Bacteroidetes and Firmicutes and the archaeal phylum Euryarchaeota. At the genus level, Prevotella (15-17%) and Methanobrevibacter (96%) dominated the bacterial and archaeal communities of the in vitro rumen fermenta, respectively. Supplementation of 0.1% E. faecium SROD resulted in the highest quantities of total bacteria and Ruminococcus flavefaciens, whereas 1.0% E. faecium SROD resulted in the highest contents of total fungi and Fibrobacter succinogenes. Overall, supplementation of 0.1% E. faecium SROD significantly increased the propionate and total volatile fatty acids concentrations but decreased the methane concentration while changing the microbial community abundance and composition.