Recent insight and future techniques to enhance rumen fermentation in dairy goats

Lovelia L. Manuad,이성실,이상석 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.8

Recent development of novel techniques in systems biology have been used to improve and manipulate the rumen microbial ecosystem and gain a deeper understanding of its physiological and microbiological interactions and relationships. This provided a deeper insight and understanding of the relationship and interactions between the rumen microbiome and the host animal. New high-throughput techniques have revealed that the dominance of Proteobacteria in the neonatal gut might be derived from the maternal placenta through fetal swallowing of amniotic fluid in utero, which gradually decreases in the reticulum, omasum, and abomasum with increasing age after birth. Multi “omics” technologies have also enhanced rumen fermentation and production efficiency of dairy goats using dietary interventions through greater knowledge of the links between nutrition, metabolism, and the rumen microbiome and their effect in the environment. For example, supplementation of dietary lipid, such as linseed, affects rumen fermentation by favoring the accumulation of α-linolenic acid biohydrogenation with a high correlation to the relative abundance of Fibrobacteriaceae. This provides greater resolution of the interlinkages among nutritional strategies, rumen microbes, and metabolism of the host animal that can set the foundation for new advancements in ruminant nutrition using multi ‘omics’ technologies.

Characterization, Metabolites and Gas Formation of Fumarate Reducing Bacteria Isolated from Korean Native Goat (Capra hircus coreanae)

MAMUAD LOVELIA,김선호,이성실,조광근,전체옥,이상석 한국미생물학회 2012 The journal of microbiology Vol.50 No.6

Fumarate reducing bacteria, able to convert fumarate to succinate, are possible to use for the methane reduction in rumen because they can compete for H2 with methanogens. In this, we isolated fumarate reducing bacteria from a rumen of Korean native goat and characterized their molecular properties [fumarate reductase A gene (frdA)], fumarate reductase activities, and productions of volatile fatty acids and gas. Eight fumarate reducing bacteria belonging to Firmicutes were isolated from rumen fluid samples of slaughtered Korean black goats and characterized their phylogenetic positions based on 16S rRNA gene sequences. PCR based analyses showed that only one strain, closely related to Mitsuokella jalaludinii, harbored frdA. The growths of M. jalaludinii and Veillonella parvula strains were tested for different media. Interestingly, M. jalaludinii grew very well in the presence of hydrogen alone, while V. parvula grew well in response of fumarate and fumarate plus hydrogen. M. jalaludinii produced higher levels of lactate (P≤0.05) than did V. parvula. Additionally, M. jalaludinii produced acetate, but not butyrate, whereas V. parvula produced butyrate, not acetate. The fumarate reductase activities of M. jalaludinii and V. parvula were 16.8 ± 0.34 and 16.9 ± 1.21 mmol NADH oxidized/min/ mg of cellular N, respectively. In conclusion, this showed that M. jalaludinii can be used as an efficient methane reducing agent in rumen.

Effect of Fumarate Reducing Bacteria on In Vitro Rumen Fermentation, Methane Mitigation and Microbial Diversity

MAMUAD LOVELIA,김선호,정창대,최연재,전체옥,이상석 한국미생물학회 2014 The journal of microbiology Vol.52 No.2

The metabolic pathways involved in hydrogen (H2) production,utilization and the activity of methanogens are theimportant factors that should be considered in controllingmethane (CH4) emissions by ruminants. H2 as one of themajor substrate for CH4 production is therefore should becontrolled. One of the strategies on reducing CH4 is throughthe use of hydrogenotrophic microorganisms such as fumaratereducing bacteria. This study determined the effect offumarate reducing bacteria, Mitsuokella jalaludinii, supplementationon in vitro rumen fermentation, CH4 production,diversity and quantity. M. jalaludinii significantly reducedCH4 at 48 and 72 h of incubation and significantly increasedsuccinate at 24 h. Although not significantly different, propionatewas found to be highest in treatment containing M. jalaludinii at 12 and 48 h of incubation. These results suggestthat supplementation of fumarate reducing bacteria toruminal fermentation reduces CH4 production and quantity,increases succinate and changes the rumen microbialdiversity.

High level of estradiol in a Holstein-Friesian cow with sacroiliac luxation after calving

Mamuad, Lovelia L.,Seo, Bosung,Kang, Seogjin,Cho, Yong-Il The Korean Society of Veterinary Science 2020 大韓獸醫學會誌 Vol.60 No.4

This is the report of sacroiliac luxation in a Holstein-Friesian cow, describing the progression of the sacroiliac luxation from the onset of its development with changes in estradiol (E2) and serum chemistry. The high estrogen concentration was assumed to be a predisposing factor of the disease that relaxed the pelvic muscles and a secondary slippery finally caused the sacroiliac luxation. The E2 was present at levels four times higher in the cow with sacroiliac luxation (167 pg/mL) than in normal cows. Above normal levels of creatinine kinase, aspartate aminotransferase, and lactate dehydrogenase were also observed in cow with sacroiliac luxation.

Effect of γ-aminobutyric acid producing bacteria on in vitro rumen fermentation, growth performance, and meat quality of Hanwoo steers

Mamuad Lovelia L.,김선호,Ku Min Jung,이상석 아세아·태평양축산학회 2020 Animal Bioscience Vol.33 No.7

Objective: The present study aimed to evaluate the effects of γ-aminobutyric acid (GABA)-producing bacteria (GPB) on in vitro rumen fermentation and on the growth performance and meat quality of Hanwoo steers. Methods: The effects of GPB (Lactobacillus brevis YM 3-30)-produced and commercially available GABA were investigated using in vitro rumen fermentation. Using soybean meal as a substrate, either GPB-produced or commercially available GABA were added to the in vitro rumen fermentation bottles, as follows: control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB; T3, 2 g/L autoclaved GPB; T4, 5 g/L autoclaved GPB; T5, 2 g/L GABA; and T6, 5 g/L GABA. In addition, 27 Hanwoo steers (602.06±10.13 kg) were subjected to a 129-day feeding trial, during which they were fed daily with a commercially available total mixed ration that was supplemented with different amounts of GPB-produced GABA (control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB). The degree of marbling was assessed using the nine-point beef marbling standard while endotoxin was analyzed using a Chromo-Limulus amebocyte lysate test. Results: In regard to in vitro rumen fermentation, the addition of GPB-produced GABA failed to significantly affect pH or total gas production but did increase the ammonia nitrogen (NH3-N) concentration (p<0.05) and reduce total biogenic amines (p<0.05). Animals fed the GPB-produced GABA diet exhibited significantly lower levels of blood endotoxins than control animals and yielded comparable average daily gain, feed conversion ratio, and beef marbling scores. Conclusion: The addition of GPB improved in vitro fermentation by reducing biogenic amine production and by increasing both antioxidant activity and NH3-N production. Moreover, it also reduced the blood endotoxin levels of Hanwoo steers.

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

정창대,Lovelia L. Mamuade,김선호,최연재,Alvin P. Soriano,조광근,전체옥,이성실,이상석 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.1

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.

Effect of Lactobacillus mucosae on In vitro Rumen Fermentation Characteristics of Dried Brewers Grain, Methane Production and Bacterial Diversity

Soriano, Alvin P.,Mamuad, Lovelia L.,Kim, Seon-Ho,Choi, Yeon Jae,Jeong, Chang Dae,Bae, Gui Seck,Chang, Moon Baek,Lee, Sang Suk Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.11

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 ($NH_3$-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 ($CH_4$) production and carbon dioxide ($CO_2$) 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 $CH_4$ and $CO_2$ in vitro.

막걸리주정박첨가 섬유질 배합사료가 반추위 발효 및육성단계 한우 거세우의 성장에 미치는 효과

정창대 ( Chang-dae Jeong ),( Lovelia L Mamuad ),고종열 ( Jong Youl Ko ),성하균 ( Ha Guyn Sung ),박근규 ( Keun Kyu Park ),이유경 ( Yoo Kyung Lee ),이상석 ( Sang-suk Lee ) 한국동물자원과학회(구 한국축산학회) 2016 축산기술과 산업 Vol.7 No.1

본 연구의 목적은 막걸리주정박을 섬유질배합사료에 첨가하였을 때 반추위 in vitro 발효성상과 한우 거세우의 성장에 미치는 효과를 조사하는데 있다. In vitro 반추위 발효성상 분석을 위해 대조구(무첨가막걸리주정박+섬유질배합사료), 처리구1 (10% 막걸리주정박+섬유질배합사료), 처리구2 (15% 막걸리주정박+섬유질배합사료)로 분류하였다. 반추위액 채취는 3마리의 한우 거세우에서 채취하였으며, 대조구와 처리구 TMR을 각각 2 g (DM)을 첨가한 후 각각 0, 12, 24 및 48시간에 시료를 채취하였다. 사양실험 연구를 위해 육성단계(6개월령) 한우 거세우 27두(196±8.66 kg)를 공시하여 3처리구로 분류하였으며, 24주 동안 체중, 사료섭취량 및 혈액특성을 조사하였다. In vitro 발효성상 결과, pH는 발효 12시간에 주정박첨가구가 대조구 보다 낮았으나, 총 가스 생성량과 암모니아 농도는 막걸리주정박 첨가에 따른 변화는 없었다. 총 휘발성 지방산은 24시간 발효 후 처리구가 대조구보다 높았으나, 다른 발효시간에서는 차이가 없었다. 사양실험결과에서 전체사료비는 10% 막걸리를 첨가시 8%의 사료비가 저감되었으며, 15% 첨가시 15%의 사료비를 줄일 수 있었다. 혈액분석 결과에서는 혈중알코올이 검출되지 않았으며, 총 증체량 및 일당증체량이 막걸리첨가구가 높았다. 본 연구의 전반적인 결과로 15% 막걸리주정박을 섬유질 배합사료에 첨가하였을 때 사료비 절감과 사료대체원료로써 사용이 가능할 것으로 보인다.

RESEARCH : Open Access ; Rumen fermentation and performance of Hanwoo steers fed total mixed ration with Korean rice wine residue

( Chang-dae Jeong ),( Lovelia L. Mamuad ),( Jong Youl Ko ),( Ha Guyn Sung ),( Keun Kyu Park ),( Yoo Kyung Lee ),( Sang-suk Lee ) 한국동물자원과학회(구 한국축산학회) 2016 한국축산학회지 Vol.58 No.1

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.

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

Kim, Seon-Ho,Mamuad, Lovelia L.,Jeong, Chang-Dae,Choi, Yeon-Jae,Lee, Sung Sill,Ko, Jong-Youl,Lee, Sang-Suk Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.12

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.