The objective of this work was to isolate a microorganism, able to produce high lactate dehydrogenase (LDH) activity, for use as a microbial feed additive. The LDH is an important enzyme for lactate conversion in the rumen, thereby possibly overcoming lactic acidosis owing to sudden increases of cereal in the diets of ruminants. In the present study, various bacterial strains were screened from a variety of environments. Among the isolated microorganisms, strain FFy 111-1 isolated from a Korean traditional fermented vegetable food called Kimchi showed the highest enzyme activity, along with retaining strong enzyme activity even in rumen fluid in vitro. Based on morphological and biochemical characteristics as well as compositions of cellular fatty acids plus API analyses, this strain was identified as Lactobacillus sp. The optimum temperature and pH for growth were found to be 30$^{\circ}C$ and pH 6.5, respectively. A maximum cell growth of 2.2 at $A_{650}$ together with LDH activity of 2.08 U per mL was achieved after 24 h of incubation. Initial characterization of FFy 111-1 suggested that it could be a potential candidate for use as a direct-fed microbial in the ruminant animals.
An experiment was conducted to study the effect of ionophore enriched cold processed mineral block supplemented with urea molasses on microbial growth and rumen fermentation. Twelve adult male crossbred cattle were divided into four groups on body weight basis. Animals were given wheat straw as a basal diet. The animals of group I and II were supplemented with concentrate mixture and animals of group III and IV were supplemented with cold processed urea molasses mineral block (UMMB). Thirty mg monensin/day/animal were supplemented to the animals of group II and 35 ppm monensin were incorporated in the UMMB supplemented to the animals of group IV. Dry matter (DM) intake did not differ significantly among groups. Mean rumen pH was higher in UMMB fed animals. Total volatile fatty acids (TVFA) concentration (mmole/L strained rumen liquor (SRL) in group III (113.19) was significantly (p<0.05) higher than those of group I (105.83) and II (108.74) but similar to group IV (109.34). TVFA production (mole/day) was similar in all the groups. The molar proportion of acetate was significantly (p<0.01) higher in the group I (59.56) than those of group II (51.73) and IV (55.91) but similar to group III (57.12). The molar proportion of propionate was significantly (p<0.01) higher in the monensin treated groups i.e. group II (38.38) and IV (36.26) than those of group I (27.78) and III (33.06). Butyrate molar percent was significantly (p<0.01) higher in group I (12.65) than those of group II (10.19), group III (9.83) and IV (7.84). The reduction of acetate and butyrate was due to UMMB and monensin resulted in lower A:P ratio. Average bacterial pool and bacterial production rate did not differ significantly among groups. Total N concentration (mg/100 ml SRL) was significantly (p<0.01) higher in the group I (55.30) and III (57.70) as compared to the group II (47.97) and IV (47.59). Ammonia-N concentration (mg/100 ml SRL) of group III (34.99) was significantly (p<0.01) higher than that of the group I (25.76) which was again significantly (p<0.01) higher than that of the group II (20.79) and IV (19.83) indicating slower release of ammonia due to monensin in diet. Total bacterial, cellulolytic, proteolytic bacterial and fungal count at 4 h post feeding did not differ significantly (p<0.05) among treatment groups. However, methanogenic bacterial count was significantly (p<0.01) higher in the group I (11.80) compared to the group II (8.43) which was significantly (p<0.01) higher than that of the group III (4.70) and IV (2.90). Average protozoal population was affected by both treatments. Thus feeding of UMMB and monensin in diet affected the rumen fermentation pattern towards propionate production, slower release of ammonia and reduction in methanogenic bacteria in the rumen.
Piao, Min Yu;Kim, Hyun-J.;Seo, J.K.;Park, T.S.;Yoon, J.S.;Kim, K.H.;Ha, Jong-K.
Asian-Australasian Journal of Animal Sciences
/
v.25
no.11
/
pp.1568-1574
/
2012
Three Holstein steers in the growing phase, each with a ruminal cannula, were used to test the hypothesis that the synchronization of the hourly rate of carbohydrate and nitrogen (N) released in the rumen would increase the amount of retained nitrogen for growth and thus improve the efficiency of microbial protein synthesis (EMPS). In Experiment 1, in situ degradability coefficients of carbohydrate and N in feeds including Korean rice wine residue (RWR) were determined. In Experiment 2, three total mixed ration (TMR) diets having different rates of carbohydrate and N release in the rumen were formulated using the in situ degradability of the feeds. All diets were made to contain similar contents of crude protein (CP) and neutral detergent fiber (NDF) but varied in their hourly pattern of nutrient release. The synchrony index of the three TMRs was 0.51 (LS), 0.77 (MS) and 0.95 (HS), respectively. The diets were fed at a restricted level (2% of the animal's body weight) in a $3{\times}3$ Latin-square design. Synchronizing the hourly supply of energy and N in the rumen did not significantly alter the digestibility of dry matter, organic matter, crude protein, NDF or acid detergent fiber (ADF) (p>0.05). The ruminal $NH_3$-N content of the LS group at three hours after feeding was significantly higher (p<0.05) than that of the other groups; however, the mean values of ruminal $NH_3$-N, pH and VFA concentration among the three groups were not significantly different (p>0.05). In addition, the purine derivative (PD) excretion in urine and microbial-N production (MN) among the three groups were not significantly different (p>0.05). In conclusion, synchronizing dietary energy and N supply to the rumen did not have a major effect on nutrient digestion or microbial protein synthesis (MPS) in Holstein steers.
Objective: This study investigated the effects of feeding anthocyanin-rich black cane treated with ferrous sulfate and molasses on animal performance, rumen fermentation, microbial composition, blood biochemical indices, and carcass characteristics in meat goats. Methods: Thirty-two Thai-native×Anglo-Nubian crossbred male goats (14.47±2.3 kg) were divided equally into two groups (n = 16) to investigate the effect of feeding diet containing 50% untreated anthocyanin-rich black cane silage (BS) vs diet containing anthocyaninrich black cane silage treated with 0.03% ferrous sulfate and 4% molasses (TBS) on average daily gain (ADG) and dry matter intake (DMI). At the end of 90 d feeding trial, the goats were slaughtered to determine blood biochemical indices, rumen fermentation, microbial composition, and carcass characteristics differences between the two dietary groups. Results: Goats fed the TBS diet had greater ADG and ADG to DMI ratio (p<0.05). TBS diet did not affect rumen fluid pH; however, goats in the TBS group had lower rumen ammonia N levels (p<0.05) and higher total volatile fatty acid concentrations (p<0.05). Goats in the TBS group had a higher (p<0.05) concentration of Ruminococcus albus but a lower (p<0.05) concentration of methanogenic bacteria. The TBS diet also resulted in lower (p<0.05) thiobarbituric acid-reactive substances concentration but higher (p<0.05) total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase concentrations in blood plasma, while having no effect on plasma protein, glucose, lipid, immunoglobin G, alanine transaminase, and aspartate aminotransferase. Meat from goats fed the TBS diet contained more intramuscular fat (p<0.05) and was more tender (p<0.05). Conclusion: In comparison to goats fed a diet containing 50% untreated anthocyanin-rich black cane silage, feeding a diet containing 50% anthocyanin-rich black cane silage treated with 0.03% ferrous sulfate and 4% molasses improved rumen fermentation and reduced oxidative stress, resulting in higher growth and more tender meat.
This experiment was conducted to investigate effects of non-ionic or zwitterionic (+/-) surfactants on digestibility of rice straw, and changes of growth of rumen mixed microbes, pH, and gas production during in vitro fermentation. Also, during in vitro ruminal fermentation, microbial attachment on rice straw was investigated using scanning electron microscopy (SEM). Tween 80 or SOLFA-850 for non-ionic surfactant (NIS), and 3-(Dodecyldimethylammonio) propanesulfanate (DDAP) for zwitterionic surfactant (ZIS) was supplemented by 0.05% and 0.1% in Dehority's artificial medium containing Holtein rumen fluid, respectively, and the substrate for fermentation was rice straw passed through 1 mm screen. The experiment was composed of 7 treatments (two levels of two NISs, two levels of a ZIS) including the control, and 6, 12, 24, 48 and 72 hr of fermentation time with 3 replications per treatment. Treatment of Tween 80 increased in vitro DM digestibilities during 48 hr and 72 h post fermentations compared to the other treatments, whereas treatment of DDAP as a ZIS resulted in decreased DM digestibility than that of the control from 24 hr post fermentation (P<0.05). Gas production in vitro was greater (P<0.05) with addition of NIS than the control or ZIS, and increased as fermentation time elapsed. Rumen mixed microbial growth was greatest with addition of Tween 80 as NIS, and lowest when DDAP as ZIS was supplemented to the fermentation tube (P<0.05). In SEM observation, rumen microbial population attached on rice straw particle was greater with addition of NIS, but was less with addition ZIS compared with the control. In conclusion we could not found any positive effects of ZIS surfactants on rumunal fermentation characteristics and rumen microbial growth rates.
Fariani, Armina;Warly, L.;Ichinohe, T.;Fujihara, T.;Harumoto, T.
Asian-Australasian Journal of Animal Sciences
/
v.9
no.3
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pp.247-254
/
1996
Three stages of growth of Italian ryegrass (pre-blooming, P-B; early-blooming, E-B; and late-blooming, L-B) were used to evaluate the effect of maturity on in vitro digestion of dry matter, fiber components and gas production. The rumen digestibility and gas production values were obtained by incubation of each sample in the rumen fluid of sheep for 12, 24, 36, 48 and 72 hr, respectively. The results showed that digestibility of dry matter (DM) significantly reduced (p < 0.05) as advancing maturity of the grass. Similarly, the digestibility of neutral detergent fiber (NDF) and acid detergent fiber (ADF) also significantly decreased (p < 0.05) with advancing maturity at all incubation times. However, the effect of maturity on digestibility of cellulose and hemicellulose was only detected when the samples were incubated more than 36 hr, where L-B was lower than P-B and E-B. Potential digestibility of nutrients, the maximum digestibility attainable in the rumen theoretically, was also higher at P-B than those of E-B and L-B. The amount of gas produced by microbial fermentation was closely related to the extent of DM digestion, and it was negatively correlated with advancing maturity of the grass.
In order to investigate the effects of supplemental ionic surfactants in in vitro ruminal fermentation, N-Lauroylsarcosine sodium salt(N-LSS) and sodium dodecyl sulfate(SDS) for negative(-) ionic surfactant, and hexadecylpyridinium chloride monohydrate(HPCM) and hexadecyltrimethyl ammonium bromide(HTAB) for positive (+) ionic surfactant were supplemented by 0.05% and 0.1% into the Dehority’s artificial medium containing rice straw(1mm) as a substrate. In vitro DM digestibility, the growth of rumen mixed microbes, pH, cumulative gas production and SEM(Scanning Electron Microscopy) observation of microbial attachment on rice straw particle were investigated through the experiment composing 9 treatments (two supplemental levels of two positive ionic(+) surfactant, two supplemental levels of two negative(-) ionic surfactant) including the control. The sample collection was at 6, 12, 24, 48 and 72 h post fermentation with 3 replications per treatments. DM digestibility in treatments supplemented (+) or (-) surfactants almost stopped afterward 12 h fermentation, in vitro DM digestibility at 72 h post fermentation in the ionic surfactants was at half level of that of the control(P<0.05). Accumulative gas production in in vitro was less(P<0.05) with addition of ionic surfactants compared to the control. The amount of rumen mixed microbes recovered from in vitro incubation fluid pleateaued at 12 h post fermentation for the positive (+) ionic surfactants, but steadily increased as fermentation time elapsed for the control. Rumen microbial growth rate was significantly(P<0.05) low in the negative(-) ionic surfactant compared to the control. pH of the incubation fluid was ranged from 6.02 to 7.20, and was the highest in the negative(-) ionic surfactants, and was the lowest in the control(P<0.05). In SEM observation, rumen microbial population attached on rice straw particle was less with addition of ionic surfactants than the control. In conclusion we could not found any positive effects of negative- and positive- charged surfactants on rumunal fermentation characteristics and rumen microbial growth rates.
In order to clarify the inhibitory effects of orchardgrass (Dactylis glomerata L.) lipids on ruminal fermentation and digestion, two experiments were carried out in vitro. Experiment 1 was carried out using residues of grass hay from which the lipid fraction was removed by ether extraction. To ground grass samples were added 0, 1.5, 3.0, 4.5 and 6.0% lipids and incubated anaerobically at $39^{\circ}C$ for 24 h, with the mixtures of artificial saliva and rumen fluid. Increasing grass lipid levels remarkably reduced DM and NDF disappearances. Volatile fatty acid concentration was significantly reduced at 3.0, 4.5 and 6.0% lipid levels. Microbial nitrogen proportion to total nitrogen tended to decrease by the addition of the lipids. These results indicated that grass lipids have a marked inhibitory effect on ruminal fermentation and digestion, especially when to the substrate was added 3% or more grass lipids as ether extracts. Experiment 2 was conducted to study the relationship between changes in the free fatty acids and changes in the fermentation traits. Samples were incubated for 3, 6, 9, 12, 15, 18, 21 and 24 h as a sole substrate. The polyunsaturated fatty acids steadily decreased during incubation, whereas the saturated fatty acid ($C_{18:0}$) increased. It was suggested that the hydrogenation was extended during the initial stage of incubation. The unsaturated fatty acids ($C_{18:2}$, $C_{18:3}$) produced at the initial stage of incubation were negatively correlated with the amount of microbial N and DM disappearance, indicating that polyunsaturated fatty acids had the possibility to show an inhibiting effect on ruminal fermentation and digestion.
The objective of this study was to investigate antimicrobial activity, during the storage period, of animal feed and any effects on in vitro rumen digestion by supplementing different levels (5.55, 11.1, and 22.2 g/kg) of freeze dried citrus peel (FDCP) to the feed compared to untreated feed and feed treated with an antifungal agent (AA) at 0.05 g/kg. In a preservation test, feed supplemented with FDCP showed no deterioration over 21 days. Untreated feed and AA-treated feed, however, showed signs of deterioration after 16 days storage. Yellow colour and red colour, measured by spectro chromameter, decreased in the untreated and AA-treated feeds, but not in feed supplemented with FDCP. Aflatoxin was detected in untreated and AA-treated feeds at 16 days (8 ppb and 2 ppb) and 21 days (8 ppb and 4 ppb), but aflatoxin was not detected in the feed supplemented with FDCP. In a second experiment, fermentation by rumen microorganisms of FDCP (22.2 g/kg) and AA (0.05 g/kg) supplemented feeds was studied in vitro. Feeds were incubated with buffered rumen fluid for 3, 6, 9, 12, 24, and 48 h. Dry matter digestibility (DMD) and organic matter digestibility (OMD) were affected by treatment, but ammonia-N, total, and individual volatile fatty acids (VFA) were not adversely affected by treatment. In conclusion, the results indicated that FDCP might be useful for inhibiting microbial growth of animal feed during storage without disrupting rumen fermentation.
Lee, Shin Ja;Shin, Nyeon Hak;Jeong, Jin Suk;Kim, Eun Tae;Lee, Su Kyoung;Lee, Il Dong;Lee, Sung Sill
Asian-Australasian Journal of Animal Sciences
/
v.31
no.1
/
pp.71-79
/
2018
Objective: Gelidium amansii (Lamouroux) is a red alga belonging to the family Gelidaceae and is commonly found in the shallow coasts of many East Asian countries, including Korea, China, and Japan. G. amansii has traditionally been utilized as an edible alga, and has various biological activities. The objective of this study was to determine whether dietary supplementation of G. amansii could be useful for improving ruminal fermentation. Methods: As assessed by in vitro fermentation parameters such as pH, total gas, volatile fatty acid (VFA) production, gas profile (methane, carbon dioxide, hydrogen, and ammonia), and microbial growth rate was compared to a basal diet with timothy hay. Cannulated Holstein cows were used as rumen fluid donors and 15 mL rumen fluid: buffer (1:2) was incubated for up to 72 h with four treatments with three replicates. The treatments were: control (timothy only), basal diet with 1% G. amansii extract, basal diet with 3% G. amansii extract, and basal diet with 5% G. amansii extract. Results: Overall, the results of our study indicate that G. amansii supplementation is potentially useful for improving ruminant growth performance, via increased total gas and VFA production, but does come with some undesirable effects, such as increasing pH, ammonia concentration, and methane production. In particular, real-time polymerase chain reaction indicated that the methanogenic archaea and Fibrobacter succinogenes populations were significantly reduced, while the Ruminococcus flavefaciens populations were significantly increased at 24 h, when supplemented with G. amansii extracts as compared with controls. Conclusion: More research is required to elucidate what G. amansii supplementation can do to improve growth performance, and its effect on methane production in ruminants.
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