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 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.

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.

Comparative Study on Calcium ion Sources in the Removal of Oxalate in Wastewater by Fluidized Bed Process

Roselle Y. Mamuad,Ming-Chun Lu,Alvin R. Caparanga 대한환경공학회 2015 대한환경공학회 학술발표논문집 Vol.2015 No.10

Effects of using different roughages in the total mixed ration inoculated with or without coculture of Lactobacillus acidophilus and Bacillus subtilis on in vitro rumen fermentation and microbial population

MIGUEL MICHELLE,MAMUAD LOVELIA,소니라모스,Ku Min Jung,정창대,김선호,조용일,이상석 아세아·태평양축산학회 2021 Animal Bioscience Vol.34 No.4

Objective: This study aimed to determine the effects of different roughages in total mixed ration (TMR) inoculated with or without coculture of Lactobacillus acidophilus (L. acidophilus) and Bacillus subtilis (B. subtilis) on in vitro rumen fermentation and microbial population. Methods: Three TMRs formulations composed of different forages were used and each TMR was grouped into two treatments: non-fermented TMR and fermented TMR (F-TMR) (inoculated with coculture of L. acidophilus and B. subtilis). After fermentation, the fermentation, chemical and microbial profile of the TMRs were determined. The treatments were used for in vitro rumen fermentation to determine total gas production, pH, ammonianitrogen (NH3-N), and volatile fatty acids (VFA). Microbial populations were determined by quantitative real-time polymerase chain reaction (PCR). All data were analyzed as a 3×2 factorial arrangement design using the MIXED procedure of Statistical Analysis Systems. Results: Changes in the fermentation (pH, lactate, acetate, propionate, and NH3-N) and chemical composition (moisture, crude protein, crude fiber, and ash) were observed. For in vitro rumen fermentation, lower rumen pH, higher acetate, propionate, and total VFA content were observed in the F-TMR group after 24 h incubation (p<0.05). F-TMR group had higher acetate concentration compared with the non-fermented group. Total VFA was highest (p<0.05) in F-TMR containing combined forage of domestic and imported source (F-CF) and F-TMR containing Italian ryegrass silage and corn silage (F-IRS-CS) than that of TMR diet containing oat, timothy, and alfalfa hay. The microbial population was not affected by the different TMR diets. Conclusion: The use of Italian ryegrass silage and corn silage, as well as the inoculation of coculture of L. acidophilus and B. subtilis, in the TMR caused changes in the pH, lactate and acetate concentrations, and chemical composition of experimental diets. In addition, F-TMR composed with Italian ryegrass silage and corn silage altered ruminal pH and VFA concentrations during in vitro rumen fermentation experiment. Objective: This study aimed to determine the effects of different roughages in total mixed ration (TMR) inoculated with or without coculture of <i>Lactobacillus acidophilus</i> (<i>L. acidophilus</i>) and <i>Bacillus subtilis</i> (<i>B. subtilis</i>) on <i>in vitro</i> rumen fermentation and microbial population.Methods: Three TMRs formulations composed of different forages were used and each TMR was grouped into two treatments: non-fermented TMR and fermented TMR (F-TMR) (inoculated with coculture of <i>L. acidophilus</i> and <i>B. subtilis</i>). After fermentation, the fermentation, chemical and microbial profile of the TMRs were determined. The treatments were used for <i>in vitro</i> rumen fermentation to determine total gas production, pH, ammonianitrogen (NH₃-N), and volatile fatty acids (VFA). Microbial populations were determined by quantitative real-time polymerase chain reaction (PCR). All data were analyzed as a 3×2 factorial arrangement design using the MIXED procedure of Statistical Analysis Systems.Results: Changes in the fermentation (pH, lactate, acetate, propionate, and NH₃-N) and chemical composition (moisture, crude protein, crude fiber, and ash) were observed. For <i>in vitro</i> rumen fermentation, lower rumen pH, higher acetate, propionate, and total VFA content were observed in the F-TMR group after 24 h incubation (p<0.05). F-TMR group had higher acetate concentration compared with the non-fermented group. Total VFA was highest (p<0.05) in F-TMR containing combined forage of domestic and imported source (F-CF) and F-TMR containing Italian ryegrass silage and corn silage (F-IRS-CS) than that of TMR diet containing oat, timothy, and alfalfa hay. The microbial population was not affected by the different TMR diets.Conclusion: The use of Italian ryegrass silage and corn silage, as well as the inoculation of coculture of <i>L. acidophilus</i> and <i>B. subtilis</i>, in the TMR caused changes in the pH, lactate and acetate concentrations, and chemical composition of experimental diets. In addition, F-TMR composed with Italian ryegrass silage and corn silage altered ruminal pH and VFA concentrations during <i>in vitro</i> rumen fermentation experiment.

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.

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

Jeong, Chang-Dae,Mamuad, Lovelia L.,Kim, Seon-Ho,Choi, Yeon Jae,Soriano, Alvin P.,Cho, Kwang Keun,Jeon, Che-Ok,Lee, Sung Sil,Lee, Sang-Suk Asian Australasian Association of Animal Productio 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 $NH_3$-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 $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ 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, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

The Effects of Total Mixed Ration Feeding with High Roughage Content on Growth Performance, Carcass Characteristics, and Meat Quality of Hanwoo Steers

Ku, Min Jung,Mamuad, Lovelia,Nam, Ki Chang,Cho, Yong Il,Kim, Seon Ho,Choi, Young Sun,Lee, Sang Suk Korean Society for Food Science of Animal Resource 2021 한국축산식품학회지 Vol.41 No.1

This study investigated the dietary effect of total mixed ration (TMR) based on high roughage content on the growth performance, carcass characteristics, and meat quality of Hanwoo steers. Twenty-four Hanwoo steers (average body weight, 195.3±4.7 kg; age, 8.5 mon) were randomly allocated to three experimental groups according to forage and concentrate ratio (DM basis): 25:75 (control), 50:50 (T50), and 70:30 (T70). Productivity in the fattening period and final body weight were significantly higher in the control. Average daily gain and feed conversion ratio were the same among treatments. Serum parameters, cholesterol, blood urea nitrogen, and total protein were higher in the control. Carcass weight was comparable in the control and T50 but feeding more roughage was significantly correlated with a higher intramuscular fat. Shear strength and drip loss were higher while n-6/n-3 was lower in T70 compared to the other groups. However, meat color was not significantly different among treatments. In terms of free amino acid contents, glutamic acid and glycine were higher in the control than T50 and T70. Overall, feeding Hanwoo steers with high forage content TMR had the lowest n-6/n-3 ratio of fatty acid content but highest intramuscular fat, shear strength, and drip loss. High forage content TMR is the best feed for Hanwoo steers that gives more benefits for human health and consumption but also provides the best meat grade and quality, which is important in the beef market in Korea.

Effects of reductive acetogenic bacteria and lauric acid on in vivo ruminal fermentation, microbial populations, and methane mitigation in Hanwoo steers in South Korea

Kim, Seon-Ho,Mamuad, Lovelia L,Choi, Yeon-Jae,Sung, Haguyn G,Cho, Kwang-Keun,Lee, Sang Suk Oxford University Press 2018 Journal of animal science Vol.96 No.11