Young Kyoon Oh

Comparative Survey of Rumen Microbial Communities and Metabolites across One Caprine and Three Bovine Groups, Using Bar-Coded Pyrosequencing and 1H Nuclear Magnetic Resonance Spectroscopy

ABSTRACT Pyrosequencing of 16S rRNA genes (targeting Bacteria and Archaea) and 1H nuclear magnetic resonance were applied to investigate the rumen microbiota and metabolites of Hanwoo steers in the growth stage (HGS), Hanwoo steers in the late fattening stage (HFS), Holstein-Friesian dairy cattle (HDC), and Korean native goats (KNG) in the late fattening stage. This was a two-part investigation. We began by comparing metabolites and microbiota of Hanwoo steers at two stages of husbandry. Statistical comparisons of metabolites and microbial communities showed no significant differences between HFS and HGS (differing by a dietary shift at 24 months and age [67 months versus 12 months]). We then augmented the study by extending the investigation to HDC and KNG. Overall, pyrosequencing of 16S rRNA genes showed that the rumens had highly diverse microbial communities containing many previously undescribed microorganisms. Bioinformatic analysis revealed that the bacterial sequences were predominantly affiliated with four phyla—Bacteroidetes, Firmicutes, Fibrobacteres, and Proteobacteria—in all ruminants. However, interestingly, the bacterial reads belonging to Fibrobacteres were present at a very low abundance (<0.1%) in KNG. Archaeal community analysis showed that almost all of these reads fell into a clade related to, but distinct from, known cultivated methanogens. Statistical analyses showed that the microbial communities and metabolites of KNG were clearly distinct from those of other ruminants. In addition, bacterial communities and metabolite profiles of HGS and HDC, fed similar diets, were distinctive. Our data indicate that bovine host breeds override diet as the key factor that determines bacterial community and metabolite profiles in the rumen.

Green synthesis of silver nanoparticles using Rheum palmatum root extract and their antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa

Abstract In the present study, silver nanoparticles (AgNPs) were synthesized by using aqueous root extracts of Rheum palmatum and characterized by various spectroscopic methods. The nanoparticles were found to be in hexagonal and spherical shapes. The average particle size was found to be 121 ± 2 nm with zeta potential values of −21.6 mv by dynamic light scattering (DLS) method. Gas chromatography–mass spectrometry (GC-MS) analysis of R. palmatum revealed 35 compounds. The synthesized AgNPs showed significant activity against Staphylococcus aureus and Pseudomonas aeruginosa with IC90 values of 15 μg/ml and IC50 values of 7.5 μg/ml, respectively. The protein leakage level was high and morphological changes occurred in bacteria treated with AgNPs.

Comparison of Methane Production in Korean Native Cattle(Hanwoo) Fed Different Grain Sources

Methane production during anaerobic fermentation in the rumen represents an energy loss to the host animal and induces emissions of greenhouse gases in the environment. Our study focused on comparison in methane production from growing Korean native steers fed different grain sources. Six Hanwoo steers (BW = 180.6 ± 3.1 kg) were fed, on a DM basis (TDN 2.80 kg), 40% timothy and 60% barley concentrate (Barley) or corn concentrate (Corn), respectively, based on the Korean Feeding Standards. Each period lasted 18 days including a 14-day adaptation and a 4-day measuring times. The steers were in the head hood chamber system (one cattle per chamber) during each measuring time to measure heat and methane production per day. Different grain sources did not affect digestibilities of dry matter, crude protein, crude fiber, crude fat, NDF, ADF and nitrogen-free extract. The mean methane concentrations per day were 202.0 and 177.1 ppm for Barley and Corn, respectively. Methane emission averaged 86.8 and 77.7 g/day for Barley and Corn, respectively. Methane emission factor by maintenance energy requirement for the growing steers fed barley based concentrate was higher than the steers fed corn based concentrate (Barley vs. Corn, 31.7 kg CH4 head 1 yr 1 vs. 28.4 kg CH4 head 1 yr 1 ). Thus, methane conversion rate was 0.065 (6.5%) and 0.055 (5.5%) for Barley and

Characteristics of Wet and Dried Distillers Grains on In vitro Ruminal Fermentation and Effects of Dietary Wet Distillers Grains on Performance of Hanwoo Steers

Two experiments were conducted to evaluate the nutrient composition, in vitro dry matter disappearance (IVDMD) and organic matter disappearance (IVOMD) of three kinds of distillers grains (DG); i) wet distillers grains (WDG, KRW 25/kg), ii) dried distillers grains (DDG, KRW 280/kg), iii) dried distillers grains with solubles (DDGS, KRW 270/kg) produced from tapioca 70% and rice 30%, and to evaluate dietary effects of WDG on the performance of Hanwoo steers. In Exp. 1, twelve-WDG, four-DDG and one-DDGS were collected from seven ethanol plants. Average crude protein, crude fiber, neutral detergent fiber, and acid detergent fiber of WDG, DDG, and DDGS were: 32.6%, 17.8%, 57.5%, and 30.2% for WDG, 36.7%, 13.9%, 51.4%, and 30.5% for DDG, and 31.0%, 11.9%, 40.3%, and 21.2% for DDGS (DM basis), respectively. The DDGS had a higher quantity of water-soluble fraction than WDG and DDG and showed the highest IVDMD (p<0.05) in comparison to others during the whole experimental time. The IVDMD at 0 to 12 h incubation were higher (p<0.05) in DDG than WDG, but did not show significant differences from 24 to 72 h. The same tendency was observed in IVOMD, showing that DG made from tapioca and rice (7:3) can be used as a feed ingredient for ruminants. Considering the price, WDG is a more useful feed ingredient than DDG and DDGS. In Exp. 2, 36 Hanwoo steers of 21 months (495.1±91 kg) were randomly assigned to one of three dietary treatments for 85 days; i) Control (total mixed ration, TMR), ii) WDG 10% (TMR containing 10% of WDG, as fed basis), and iii) WDG 20% (TMR containing 20% of WDG, as fed basis). With respect to body weight and average daily gain, there were no differences between control and WDG treatments during the whole experimental period. Dry matter intake of control (9.34 kg), WDG 10% (9.21 kg) and 20% (8.86 kg) and feed conversion ratio of control (13.0), WDG 10% (13.2) and 20% (12.1) did not show differences between control and WDG treatments. Thus, the use of WDG up to 20% in TMR did not show any negative effect on the performance of Hanwoo steers.

Estimation of Availability of Whole Crop Barley and Rye Silage TMR in the Cattle

The present study was conducted to examine the fermentation characteristics and effective degradability (ED) in the rumen, and whole tract digestibility of whole crop silage based TMR in comparison with conventional separate feeding of concentrate and roughage. Three ruminally fistulated non-lactating Holstein cattle were used in a 3 × 3 Latin square design. The cattle were fed 8kg of whole crop barley silage based TMR (BS-TMR) or 8kg (DM basis) of whole crop rye silage based TMR (RS-TMR) twice (08:00 and 18:00) daily in an equal amount. The cattle were also fed concentrate (5.6kg) and rice straw (1.4kg) seperately (DM basis, Control) twice daily in an equal amount. The both silages were included in TMR at 20% level (as fed basis). pH in the rumen fluid was not influenced by the diets but was slightly higher from TMR than from control. No difference was found in ammonia-N concentration between diets. Total VFA concentration was relatively increased in the cattle fed RS-TMR to the other diets up to 6h post feeding. The proportion of acetate was increased in the TMR feeding at right before feeding (0 h, p<0.005) and 9 h (p<0.048) post feeding compared with control. Propionate proportion was increased (p<0.046) in both TMRs while butyrate proportion was increased (p<0.029) at 1h post feeding compared to other diets. Effective degradability (ED) of DM and CP of RS-TMR was relatively increased to other diets, and EDNDF of both TMRs was higher than that of control diet due to the increased parameters b (p<0.039) and c (p<0.006) in TMR treatments. Whole track digestibility of most components in the TMRs was slightly increased compared to that in control diet, and RS-TMR had a tendency to be increased whole track digestibility except for NDF compared to BS-TMR. Based on the results observed from the present study, nutrient availability of whole crop silage based TMR looked slightly better than conventional separate feeding of concentrate and rice straw, mainly due to the improved stabilization of fermentation in the rumen and increased NDF digestibility of whole crop silage in TMR. (Key words : Whole crop silage, TMR, Fermentation characteristics, Effective degradability, Whole tract digestibility)

Effect of rhubarb (Rheum spp.) root on in vitro and in vivo ruminal methane production and a bacterial community analysis based on 16S rRNA sequence

The objective of this study was to evaluate the anti-methanogenic effect of rhubarb (Rheum spp.) on in vitro, in vivo, and bacterial community composition using Quantitative Insights into Microbial Ecology sequencing. Rhubarb root powder was tested at different concentrations (0, 0.33, 0.67, and 1.33 g/L) in vitro, and all incubations were carried out in triplicate two runs on separate days. Concentrations of 0.67 and 1.33 g/L rhubarb significantly (P < 0.05) reduced methane production and the acetate : propionate ratio compared with those of the Control, without adverse effects on total volatile fatty acids and total gas production. In the second in vivo trial, four Hanwoo (Korean native) steers (live bodyweight, 556 ± 46 kg) with a ruminal cannula were housed individually in metabolic stalls and fed a basal diet twice daily in equal amounts at 0900 hours and 2100 hours. The before rhubarb treatment (before treatment) duration was 24 days for all steers; 14 days were used for diet adaptation and 10 days were used for gas samples collected 1, 2, and 3 h after the morning feeding on Days 3, 5, 7, and 9. We used three syringe needles passed through the ruminal cannula stopper at different time points as a simple and rapid method to sample rumen gas. Thereafter, three mesh bags containing 30 g of sliced rhubarb root each were placed at different depths in the rumen of each steer for 14 days (after treatment), and gas samples were collected on Days 4, 7, 10, 12, and 13. The results showed a significant (P < 0.05) decrease in methane concentration from the rhubarb-treated steers and provide the evidence that this method would be useful for in vivo screening of anti-methanogenic feed additives or plant material. Furthermore, 16s RNA sequencing after treatment showed increases in the numbers of Prevotella, and Lactobacillus, but decreases in Methanobrevibacter. In conclusion, rhubarb had an anti-methanogenic effect in vitro and in vivo, and the increase in the number of Prevotella shifted ruminal fermentation towards propionate production.

Effects of Saponin Contained Plant Extracts on Ruminal Fermentation Characteristics and Methane Production

This study was conducted to evaluate the effects of saponin contained plant extracts on in vitro rumen fermentation characteristics and methane production. Ruminal fluid was collected from rumen cannulated Hanwoo steers fed rice straw and concentrate (5:5). Collected rumen fluids, corn starch and buffer including saponin contained plant extracts (ginseng, Ogapi, soapwort, tea plant and yucca; 0.5%/15 ml) were incubated at 39 for 24 h. All incubations were repeated five times. Rumen pH in all treatments was lower (p<0.05) compared with that of the control (no addition) during incubation time. The concentration of total VFA in all treatments was higher (p<0.05) than that of the control after 12h incubation. Compared with the control, the concentration of acetate and propionate in all treatments was lower and higher after 6h incubation, respectively. The concentration of NH3-N in all treatments was lower (p<0.05) than that of the control except for Ogapi or yucca extracts supplementation. The number of protozoa in all treatments was significantly (p<0.05) lower than that of the control except for soapwort extract supplementation. The total gas production and methane production in all treatments was higher (p<0.05) and lower (p<0.05) compared with the control, except for ogapi or soapwort extracts supplementation after 12h incubation, respectively. Therefore, reduction in methane production by saponins may could be results from decreased protozoal population without any negative in vitro fermentation . (Key words : Saponin, Methane, Ruminal Fermentation, Protozoa)

Effects of deoxynivalenol- and zearalenone-contaminated feed on the gene expression profiles in the kidneys of piglets.

Objective Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), common contaminants in the feed of farm animals, cause immune function impairment and organ inflammation. Consequently, the main objective of this study was to elucidate DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the kidneys of piglets. Methods Fifteen 6-week-old piglets were randomly assigned to three dietary treatments for 4 weeks: control diet, and diets contaminated with either 8 mg DON/kg feed or 0.8 mg ZEN/kg feed. Kidney samples were collected after treatment, and RNA-seq was used to investigate the effects on immune-related genes and gene networks. Results A total of 186 differentially expressed genes (DEGs) were screened (120 upregulated and 66 downregulated). Gene ontology analysis revealed that the immune response, and cellular and metabolic processes were significantly controlled by these DEGs. The inflammatory stimulation might be an effect of the following enriched Kyoto encyclopedia of genes and genomes pathway analysis found related to immune and disease responses: cytokine-cytokine receptor interaction, chemokine signaling pathway, toll-like receptor signaling pathway, systemic lupus erythematosus (SLE), tuberculosis, Epstein-Barr virus infection, and chemical carcinogenesis. The effects of DON and ZEN on genome-wide expression were assessed, and it was found that the DEGs associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9, CXCL10, chemokine [C-C motif] ligand 4), proliferation (insulin like growth factor binding protein 4, IgG heavy chain, receptor-type tyrosine-protein phosphatase C, cytochrome P450 1A1, ATP-binding cassette sub-family 8), and other immune response networks (lysozyme, complement component 4 binding protein alpha, oligoadenylate synthetase 2, signaling lymphocytic activation molecule-9, α-aminoadipic semialdehyde dehydrogenase, Ig lambda chain c region, pyruvate dehydrogenase kinase, isozyme 4, carboxylesterase 1), were suppressed by DON and ZEN. Conclusion In summary, our results indicate that high concentrations of DON and ZEN suppress the inflammatory response in kidneys, leading to potential effects on immune homeostasis.

Deoxynivalenol- and zearalenone-contaminated feeds alter gene expression profiles in the livers of piglets.

Objective The Fusarium mycotoxins of deoxynivalenol (DON) and zerolenone (ZEN) cause health hazards for both humans and farm animals. Therefore, the main intention of this study was to reveal DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the liver of piglets. Methods In the present study, 15 six-week-old piglets were randomly assigned to the following three different dietary treatments for 4 weeks: control diet, diet containing 8 mg DON/kg feed, and diet containing 0.8 mg ZEN/kg feed. After 4 weeks, liver samples were collected and sequenced using RNA-Seq to investigate the effects of the mycotoxins on genes and gene networks associated with the immune systems of the piglets. Results Our analysis identified a total of 249 differentially expressed genes (DEGs), which included 99 upregulated and 150 downregulated genes in both the DON and ZEN dietary treatment groups. After biological pathway analysis, the DEGs were determined to be significantly enriched in gene ontology terms associated with many biological pathways, including immune response and cellular and metabolic processes. Consistent with inflammatory stimulation due to the mycotoxin-contaminated diet, the following Kyoto encyclopedia of genes and genomes pathways, which were related to disease and immune responses, were found to be enriched in the DEGs: allograft rejection pathway, cell adhesion molecules, graft-versus-host disease, autoimmune thyroid disease (AITD), type I diabetes mellitus, human T-cell leukemia lymphoma virus infection, and viral carcinogenesis. Genome-wide expression analysis revealed that DON and ZEN treatments downregulated the expression of the majority of the DEGs that were associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9), proliferation (insulin-like growth factor 1, major facilitator superfamily domain containing 2A, insulin-like growth factor binding protein 2, lipase G, and salt inducible kinase 1), and other immune response networks (paired immunoglobulin-like type 2 receptor beta, Src-like-adaptor-1 [SLA1], SLA3, SLA5, SLA7, claudin 4, nicotinamide N-methyltransferase, thyrotropin-releasing hormone degrading enzyme, ubiquitin D, histone H2B type 1, and serum amyloid A). Conclusion In summary, our results demonstrated that high concentrations DON and ZEN disrupt immune-related processes in the liver.

Profile of Hanwoo Steer Carcass Characteristics, Meat Quality and Fatty Acid Composition after Feeding Italian Ryegrass Silage

The objective of this work was to evaluate the growth performance, feed intake, slaughter characteristics, meat quantity and quality characteristics of Hanwoo steers fed with Italian ryegrass (IRG) silage (TRT). IRG silage consisted 11.70% protein, 2.84% ether extract, 53.50% dry matter digestibility and 63.34% total digestible nutrients. The daily weight gain and feed conversion ratio of TRT were significantly (p<0.01) higher than that of control diet (CON; fed rice straw) in the whole periods. However, the slaughter weight, dressing percentage, quantity grade and quantity traits (marbling score, meat color, fat color, and quality grade) of either TRT or CON were similar. Meat fed TRT diet showed higher crude fat and lightness (L*) value and lower moisture content and pH value compared with the CON diet (p<0.05). Overall the carcass yield was 12.5% higher than CON diet.