Evaluation of Different Feeds for Their Potential to Generate Methane and Change Methanogen Diversity

Optimization of the dietary formulation is the most effective way to reduce methane. Nineteen feed ingredients (brans, vegetable proteins, and grains) were evaluated for their potential to generate methane and change methanogen diversity using an in vitro ruminal fermentation technique. Feed formulations categorized into high, medium and low production based on methane production of each ingredient were then subjected to in vitro fermentation to determine the real methane production and their effects on digestibility. Methanogen diversity among low, medium and high-methane producing groups was analyzed by PCR-DGGE. The highest methane production was observed in Korean wheat bran, soybean and perilla meals, and wheat and maize of brans, vegetable protein and cereal groups, respectively. On the other hand, corn bran, cotton seed meal and barley led to the lowest production in the same groups. Nine bacteria and 18 methanogen 16s rDNA PCR-DGGE dominant bands were identified with 83% to 99% and 92% to 100% similarity, respectively. Overall, the results of this study showed that methane emissions from ruminants can be mitigated through proper selection of feed ingredients to be used in the formulation of diets

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

Effect of Phytogenic Feed Additives in Soybean Meal on Swine Fermentation for Odor Reduction and Bacterial Community Comparison

The effect of different phytogenic feed additives on reducing odorous compounds in swine was investigated using in vitro fermentation and analyzed their microbial communities. Soybean meal (1%) added with 0.1% different phytogenic feed additives (FA) were in vitro fermented using swine fecal slurries and anaerobically incubated for 12 and 24 h. The phytogenic FAs used were red ginseng barn powder (Panax ginseng C. A. Meyer, FA1), persimmon leaf powder (Diospyros virginiana L., FA2), ginkgo leaf powder (Ginkgo biloba L., FA3), and oregano lippia seed oil extract (Lippia graveolens Kunth, OL, FA4). Total gas production, pH, ammonia-nitrogen (NH3-N), hydrogen sulfide (H2S), nitrite-nitrogen (NO2−-N), nitrate-nitrogen (NO3−-N), sulfate (SO4−−), volatile fatty acids (VFA) and other metabolites concentration were determined. Microbial communities were also analyzed using 16S rRNA DGGE. Results showed that the pH values on all treatments increased as incubation time became longer except for FA4 where it decreased. Moreover, FA4 incubated for 12 and 24 h was not detected in NH3-N and H2S. Addition of FAs decreased (p<0.05) propionate production but increased (p<0.05) the total VFA production. Ten 16S rRNA DGGE bands were identified which ranged from 96 to 100% identity which were mostly isolated from the intestine. Similarity index showed three clearly different clusters: I (FA2 and FA3), II (Con and FA1), and III (FA4). Dominant bands which were identified closest to Eubacterium limosum (ATCC 8486T), Uncultured bacterium clone PF6641 and Streptococcus lutetiensis (CIP 106849T) were present only in the FA4 treatment group and were not found in other groups. FA4 had a different bacterial diversity compared to control and other treatments and thus explains having lowest odorous compounds. Addition of FA4 to an enriched protein feed source for growing swine may effectively reduce odorous compounds which are typically associated with swine production

Rumen fermentation and performance of Hanwoo steers fed total mixed ration with Korean rice wine residue

Abstract Background This study was conducted to evaluate the effects of adding Korean rice wine residue (RWR) in total mixed ration (TMR) on in vitro ruminal fermentation and growth performance of growing Hanwoo steers. Methods For in vitro fermentation, the experimental treatments were Control (Con: 0 % RWR + TMR), Treatment 1 (T1: 10 % RWR + TMR), and Treatment 2 (T2: 15 % RWR + TMR). The rumen fluid was collected from three Hanwoo steers and mixed with buffer solution, after which buffered rumen fluid was transferred into serum bottles containing 2 g dry matter (DM) of TMR added with or without RWR. The samples were then incubated for 0 h, 12 h, 24 h, or 48 h at 39 °C and 100 rpm. For the in vivo experiment, 27 Hanwoo steers (6 months old) with an average weight of 196 ± 8.66 kg were subjected to a 24-week feeding trial. The animals were randomly selected and equally distributed into three groups. After which the body weight, feed intake and blood characteristics of each group were investigated. Results The pH of the treatments decreased significantly relative to the control during the 12 h of incubation. Total gas production and ammonia nitrogen (NH3-N) was not affected by RWR addition. The total volatile fatty acid (VFA) was lower after 24 h of incubation but at other incubation times, the concentration was not affected by treatments. Feed cost was 8 % and 15 % lower in T1 and T2 compared to control. Blood alcohol was not detected and a significant increase in total weight gain and average daily gain were observed in Hanwoo steers fed with RWR. Conclusion Overall, the results of this study suggest that TMR amended with 15 % RWR can be used as an alternative feed resource for ruminants to reduce feed cost

Effect of on Rumen Fermentation Characteristics of Dried Brewers Grain, Methane Production and Bacterial Diversity

The effects of Lactobacillus mucosae (L. mucosae), a potential direct fed microbial previously isolated from the rumen of Korean native goat, on the rumen fermentation profile of brewers grain were evaluated. Fermentation was conducted in serum bottles each containing 1% dry matter (DM) of the test substrate and either no L. mucosae (control), 1% 24 h broth culture of L. mucosae (T1), or 1% inoculation with the cell-free culture supernatant (T2). Each serum bottle was filled anaerobically with 100 mL of buffered rumen fluid and sealed prior to incubation for 0, 6, 12, 24, and 48 h from which fermentation parameters were monitored and the microbial diversity was evaluated. The results revealed that T1 had higher total gas production (65.00 mL) than the control (61.33 mL) and T2 (62.00 mL) (p<0.05) at 48 h. Consequently, T1 had significantly lower pH values (p<0.05) than the other groups at 48 h. Ammonia nitrogen (NH3-N), individual and total volatile fatty acids (VFA) concentration and acetate:propionate ratio were higher in T1 and T2 than the control, but T1 and T2 were comparable for these parameters. Total methane (CH4) production and carbon dioxide (CO2) were highest in T1. The percent DM and organic matter digestibilities were comparable between all groups at all times of incubation. The total bacterial population was significantly higher in T1 (p<0.05) at 24 h, but then decreased to levels comparable to the control and T2 at 48 h. The denaturing gradient gel electrophoresis profile of the total bacterial 16s rRNA showed higher similarity between T1 and T2 at 24 h and between the control and T1 at 48 h. Overall, these results suggest that addition of L. mucosae and cell-free supernatant during the in vitro fermentation of dried brewers grain increases the VFA production, but has no effect on digestibility. The addition of L. mucosae can also increase the total bacterial population, but has no significant effect on the total microbial diversity. However, inoculation of the bacterium may increase CH4 and CO2 in vitro

Bacterial Community Dynamics during Swine Fermentation Using Starch as a Substrate with Different Feed Additives for Odor Reduction

The experiment was conducted by in vitro fermentation and bacterial community analysis to investigate the reduction of odorous compounds in response to the use of feed additives (FA) during carbohydrate overload in growing pigs. Soluble starch at 1% (control) and various FA at 0.1% Ginseng meal (FA1); Persimmon leaf (FA2); Gingko nut (FA3) and Oregano lippia (FA4) were added to fecal slurry and incubated anaerobically for 12 and 24 h. In vitro parameters and microbial diversity of the dominant bacteria following fermentation were analyzed using Denaturing Gradient Gel Electrophoresis (DGGE), band cloning and sequencing of the V3 region. Results showed that total gas production increased with the advancement of incubation (p<0.05). pH values of FAs and control groups were decreased except the FA4 group which increased somewhat from 12 to 24 h (p<0.05). Ammonia nitrogen (NH3-N) and H2S gas concentrations were comparatively lower in both stages in FA4 treatment than in the other groups (p<0.05). Hence, NH3-N concentrations in liquid phases were increased (p<0.05) from 12 to 24 h, but the trend was lowest in FA4 than in the other groups at both stages. The total VFA production was comparatively lower and butyrate levels were moderate in FA4 group than in the the other groups during both stages (p<0.05). Indirect odor-reducing compounds such as NO2, NO3 and SO4 concentrations were higher in the FA4 and FA3 than in the other groups at 24 h (p<0.05). After fermentation, ten dominant bands appeared, six of which appeared in all samples and four in only the FA4 treated group. The total number of DGGE bands and diversity was higher in the FA4-group compared to other groups. Additionally, similarity indices were lowest (71%) in the FA4, which represented a different bacterial community compared with the other groups. These findings indicate that NH3-N, H2S and VFA production was minimal, and pH was also better in the FA4 group than in the other groups. Furthermore, the conversion of odor-reducing indirect compounds or their intermediates was higher in the FA4 group in compared to the other groups. FA4 group generated less odorous products and more indirect products by in vitro fermentation at 24 h, and their microbial pattern appeared to differ from that of the other groups. These findings suggest that this particular FA could change the microbial population, which may have a beneficial effect on odor reduction. It is recommended that the oregano lippia may be supplied to growing pigs as FA along with excess carbohydrate sources to reduce the production of odorous compounds

Effect of different concentrate diet levels on rumen fluid inoculum used for determination of in vitro rumen fermentation, methane concentration, and methanogen abundance and diversity

This study investigated the effects of different diet concentrate levels in animal diets as a source of rumen fluid inoculum for in vitro ruminal fermentation, CH4 concentration, and methanogen abundance and diversity. Three non-lactating mature cows assigned to a 3 × 3 Latin square design were fed with Italian rye grass and concentrate feed at 2 (low), 5 (medium), and 8 (high) kg per animal per day were used as rumen fluid donors. The experiment consisted of three 21-d periods that each consisted of 14 d adaptation followed by 7 d of continuous feeding prior to rumen fluid collection for in vitro fermentation evaluation. High concentrate proportion produced the highest total gas in all incubation periods (p < .05) while methane (CH4) concentration was highest in low concentrate proportions and lowest in medium concentrate proportions. Propionate concentration was highest in high concentrate proportions, whereas butyrate concentration was highest in medium concentrate proportions. High concentrate proportion at 0 and 12 h and medium concentrate proportions at 12 h produced archaeal denaturing gradient gel electrophoresis (DGGE) profiles that differed from those of low concentrate proportions. Medium concentrate proportion had the lowest methanogen DNA copies at 12 h of incubation (p < .05). Changes in diet influenced the rumen microbiome, CH4 concentration, and methanogen diversity and abundance in cattle. The rumen conditions of an animal, as along with the microbiome, change as the feed diet changes. As a result, the inoculum for in vitro rumen fermentation studies affects Volatile fatty acid (VFA) concentrations, CH4 production, and methanogen diversity and abundance

Use of Lysozyme as a Feed Additive on Rumen Fermentation and Methane Emission

This study was conducted to determine the effect of lysozyme addition on in vitro rumen fermentation and to identify the lysozyme inclusion rate for abating methane (CH4) production. An in vitro ruminal fermentation technique was done using a commercial concentrate to rice straw ratio of 8:2 as substrate. The following treatments were applied wherein lysozyme was added into 1 mg dry matter substrate at different levels of inclusion: Without lysozyme, 2,000, 4,000, and 8,000 U lysozyme. Results revealed that, lysozyme addition had a significant effect on pH after 24 h of incubation, with the highest pH (p<0.01) observed in 8,000 U lysozyme, followed by the 4,000 U, 2,000 U, and without lysozyme. The highest amounts of acetic acid, propionic acid (p<0.01) and total volatile fatty acid (TVFA) (p<0.05) were found in 8,000 U after 24 h of incubation. The CH4 concentration was the lowest in the 8,000 U and the highest in the without lysozyme addition after 24 h of incubation. There was no significant differences in general bacteria, methanogen, or protozoan DNA copy number. So far, addition of lysozyme increased the acetate, propionate, TVFA, and decreased CH4 concentration. These results suggest that lysozyme supplementation may improve in vitro rumen fermentation and reduce CH4 emission

Effect of γ-Aminobutyric Acid (GABA) Producing Bacteria on Rumen Fermentation, Biogenic Amine Production and Anti-oxidation Using Corn Meal as Substrate

The effects and significance of γ-amino butyric acid (GABA) producing bacteria (GPB) on in vitro rumen fermentation and reduction of biogenic amines (histamine, methylamine, ethylamine, and tyramine) using corn meal as a substrate were determined. Ruminal samples collected from ruminally fistulated Holstein cows served as inoculum and corn was used as substrate at 2% dry matter (DM). Different inclusion rates of GPB and GABA were evaluated. After incubation, addition of GPB had no significant effect on in vitro fermentation pH and total gas production, but significantly increased the ammonia nitrogen (NH3-N) concentration and reduced the total biogenic amines production (p<0.05). Furthermore, antioxidation activity was improved as indicated by the significantly higher concentration of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) among treated samples when compared to the control (p<0.05). Additionally, 0.2% GPB was established as the optimum inclusion level. Taken together, these results suggest the potential of utilizing GPB as feed additives to improve growth performance in ruminants by reducing biogenic amines and increasing anti-oxidation

Enhanced Ruminal Fermentation Parameters and Altered Rumen Bacterial Community Composition by Formulated Rumen Buffer Agents Fed to Dairy Cows with a High-Concentrate Diet

The effects of rumen buffer agents on ruminal fermentation parameters and bacterial community composition were determined using in vitro and in vivo experiments in three rumen-cannulated, high-concentrate fed Holstein Friesian dairy cows. Experiment 1 in vitro treatments included bentonite, calcium carbonate, calcium oxide, sodium bicarbonate, sodium sesquicarbonate, and processed coral, and unbuffered samples served as the control. Experiment 2 in vitro treatments were based on the formulation of various combinations of the buffer agents used in Experiment 1. Combinations were selected for the in vivo study based on their buffering ability. Calcium oxide, sodium bicarbonate, and sodium sesquicarbonate stabilized the ruminal pH and improved in vitro rumen fermentation. The combined buffer agents had a significant effect on pH, buffering capacity, total gas, and total volatile fatty acids. Firmicutes and Bacteroidetes were the dominant phyla in both treatments and the control. Ruminococcus and Prevotella were found to be the dominant genera. Ruminococcus bromii was predominant in the treatment group. Prevotella jejuni was more abundant in the control group compared to the treatment group, in which its abundance was very low. Ruminococcus flavefaciens and Intestinimonas butyriciproducens gradually increased in abundance as cows received treatment. Overall, a high-concentrate diet administered to cows induced adverse changes in ruminal pH; however, buffer supplementation enhanced ruminal fermentation characteristics and altered bacterial community, which could contribute to preventing ruminal acidosis