Effect of malate on in vitro (RUSITEC) rumen fermentation

1998 ◽  
Vol 22 ◽  
pp. 303-305 ◽  
Author(s):  
M. D. Carro ◽  
S. Lopez ◽  
C. Valdes ◽  
F. J. Ovejero
Keyword(s):  
Volatile Fatty Acids ◽  
Artificial Saliva ◽  
Rumen Fermentation ◽  
Disodium Salt ◽  
Antibiotic Resistant ◽  
Selenomonas Ruminantium ◽  
Alfalfa Hay ◽  
Micro Organisms ◽  
Electron Sink

In the last few years there has been an increasing concern regarding the use of antibiotics in ruminant feeding and the potential for selection of antibiotic-resistant pathogen micro-organisms. Some authors (Martin and Streeter, 1995; Callaway and Martin, 1996) suggested that organic acids (aspartate, fumarate, malate) potentially provide an alternative to currently used antimicrobial compounds. Several in vitro studies (Martin and Streeter, 1995; Callaway and Martin, 1996) showed that incorporation of DL-malate into fermentations with both Selenomonas ruminantium HD4 and with mixed ruminal micro-organisms resulted in changes in final pH, methane and volatile fatty acids (VFA) that are analogous to ionophore effects. Nisbet and Martin (1993) hypothesized that malate acted as an electron sink for hydrogen. However, the mechanism of action is not well known. Malate is a key intermediate in the succinate-propionate pathway and therefore could stimulate propionate production. The objective of this study was to study the effects of DL-malate and propionate on the in vitro rumen fermentation of a 50:50 foragexoncentrate diet.The study was carried out using the rumen simulation technique (RUSITEC) following the general incubation procedure described by Czerkawski and Breckenridge (1977). The complete unit consisted of eight vessels with an effective volume of 700 ml each. The vessels inocula (solid and liquid) were obtained from three ruminally fistulated ewes given a diet consisting of 700 g alfalfa hay and 300 g concentrate per kg dry matter (DM) (Table 1) and transferred to the RUSITEC system within 30 min of the 1st day of the experiment. The flow through the vessels was maintained by continuous infusion of artificial saliva at a rate of 533 ml/day. Each vessel received daily a nylon bag containing 7 g alfalfa hay, 7 g concentrate and 0·10 g vitaminsminerals mix. From the 1st day of incubation three vessels received daily 5·62 mmol DL-malate (disodium salt; Sigma-Aldrich Quimica, S.A., Spain), three vessels received 5·62 mmol of propionate (monosodium salt; Sigma-Aldrich Quimica, S.A., Spain) and two vessels received no addition (control). DL-malate and propionate were weighed into the nylon bags and carefully mixed with the food.

Effect of melamine onin vitrorumen microbial growth, methane production and fermentation of Chinese wild rye hay and maize meal in binary mixtures

2013 ◽  
Vol 152 (4) ◽  
pp. 686-696 ◽  
Author(s):  
H. J. YANG ◽  
H. ZHUANG ◽  
X. K. MENG ◽  
D. F. ZHANG ◽  
B. H. CAO
Keyword(s):  
Volatile Fatty Acids ◽  
Ammonia Nitrogen ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Hydrogen Gas ◽  
Batch Cultures ◽  
Branched Chain ◽  
Micro Organisms ◽  
High Forage

SUMMARYThe effects of melamine on gas production (GP) kinetics, methane (CH4) production and fermentation of diets differing in forage content (low-forage (LF) diet: 200 g/kg and high-forage (HF) diet: 800 g/kg) by rumen micro-organismsin vitrowere studied using batch cultures. Rumen contents were collected from three Simmental×Luxi crossbred beef cattle. Melamine was added to the incubation bottles to achieve final concentration of 0 (control), 2, 6, 18, 54, 162 and 484 mg/kg of each diet. Cumulative GP was continuously measured in an automated gas recording instrument during 72 h of incubation, while fermentation gas end-products were collected to determine molar proportions of carbon dioxide (CO2), CH4and hydrogen gas (H2) in manually operated batch cultures. Differences in GP kinetics and fermentation gases were observed in response to the nature of the diets incubated. Although melamine addition did not affect GP kinetics and fermentation gas pattern compared to the control, the increase of melamine addition stimulated the yield of CH4by decreasing CO2, especially during the fermentation of the HF diet. The concentrations of ammonia nitrogen (N), amino acid N and microbial N in culture fluids were greater in the fermentation of LF- than HF diets, and these concentrations were increased by the increase of melamine addition after 72-h fermentation. The concentrations of total volatile fatty acids (VFA) were greater in HF than LF diets. The addition of melamine decreased total VFA concentrations and this response was greater in HF than LF diet fermentations. Melamine addition did not affect molar proportions of acetate, butyrate, propionate and valerate compared with the control; however, branched-chain VFA production, which was lower in the HF than the LF diet, was increased by the melamine addition, especially in the HF diet fermentation. The ratio of non-glucogenic to glucogenic acids was lower in the HF than the LF diet, but it was not affected by melamine addition. In brief, the greater reduction in the rate and extent of rumen fermentation found for the HF diet in comparison with the LF diet suggested that rumen fermentation rate and extentin vitrodepended mainly on the nature of the incubated substrate, and that they could be further inhibited by the increase of melamine addition.

scholarly journals Effects of pre-incubation in sheep and goat saliva on in vitro rumen digestion of tanniferous browse foliage

2013 ◽  
Vol 151 (6) ◽  
pp. 898-906
Author(s):  
H. AMMAR ◽  
R. BODAS ◽  
J. S. GONZÁLEZ ◽  
A. Z. M. SALEM ◽  
F. J. GIRÁLDEZ ◽  
...  
Keyword(s):  
Rumen Fluid ◽  
Artificial Saliva ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Parotid Saliva ◽  
Rosa Canina ◽  
Fermentation Kinetics ◽  
Alfalfa Hay ◽  
Parotid Duct

SUMMARYA two-stage in vitro procedure was used for assessing the activity of parotid saliva to enhance rumen digestion of tanniniferous browse foliage. The procedure consisted of pre-incubation in saliva for 4 h at 39 °C followed by incubation in diluted buffered rumen fluid. Using this procedure, a study was conducted to examine the effects of pre-incubation in sheep (SS), quebracho-supplemented sheep (qSS) and goat (GS) parotid saliva or in McDougall's artificial saliva (AS, used as control) on in vitro rumen fermentation kinetics (estimated using the gas production technique) of browse foliage from six shrub species (Cytisus scoparius, Genista florida, Rosa canina, Quercus pyrenaica, Cistus laurifolius and Erica australis) collected over two seasons (spring and autumn), thus varying the in vitro digestibility (from 0·597 to 0·903) and tannin contents (from 3 to 130 g tannic acid equivalent/kg dry matter (DM)). Saliva was collected from four sheep and four goats fed alfalfa hay, and from four sheep fed the same alfalfa hay but supplemented with quebracho (rich in condensed tannins) for 60 d, through a cannula inserted in the parotid duct, and rumen fluid was always from sheep fed alfalfa hay. The extent of degradation when browse foliage was pre-incubated in qSS was similar to that observed with control AS (0·449 v. 0·452, respectively), and 8% less than the value with pre-incubation in SS (0·490). In vitro fermentation kinetics (gas production parameters) of browse foliage were not significantly enhanced with pre-incubation in qSS compared with SS, whereas in vitro digestibility and extent of degradation in the rumen were significantly reduced with qSS compared with SS. After pre-incubation in sheep and goat saliva, the extent of browse foliage degradation was significantly increased by 4–8% compared with pre-incubation in the control AS. Fermentation efficiency of browse foliage was increased (P<0·05) with pre-incubation in GS compared with SS. Sheep or goat saliva may have some activity to affect in vitro rumen fermentation of the foliage samples incubated, enhancing extent of degradation of tannin-rich browse. However, a relationship between the magnitude of this effect and the tannin content of the browse foliage could not be established, suggesting that sheep and goat saliva may not be particularly important in neutralizing tannins.

scholarly journals Pilot Study of the Effects of Polyphenols from Chestnut Involucre on Methane Production, Volatile Fatty Acids, and Ammonia Concentration during In Vitro Rumen Fermentation

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 108
Author(s):  
Yichong Wang ◽  
Sijiong Yu ◽  
Yang Li ◽  
Shuang Zhang ◽  
Xiaolong Qi ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Acetic Acid ◽  
Propionic Acid ◽  
Acid Content ◽  
Volatile Fatty Acids ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Quadratic Response ◽  
In Vitro Rumen Fermentation

Nutritional strategies can be employed to mitigate greenhouse emissions from ruminants. This article investigates the effects of polyphenols extracted from the involucres of Castanea mollissima Blume (PICB) on in vitro rumen fermentation. Three healthy Angus bulls (350 ± 50 kg), with permanent rumen fistula, were used as the donors of rumen fluids. A basic diet was supplemented with five doses of PICB (0%–0.5% dry matter (DM)), replicated thrice for each dose. Volatile fatty acids (VFAs), ammonia nitrogen concentration (NH3-N), and methane (CH4) yield were measured after 24 h of in vitro fermentation, and gas production was monitored for 96 h. The trial was carried out over three runs. The results showed that the addition of PICB significantly reduced NH3-N (p < 0.05) compared to control. The 0.1%–0.4% PICB significantly decreased acetic acid content (p < 0.05). Addition of 0.2% and 0.3% PICB significantly increased the propionic acid content (p < 0.05) and reduced the acetic acid/propionic acid ratio, CH4 content, and yield (p < 0.05). A highly significant quadratic response was shown, with increasing PICB levels for all the parameters abovementioned (p < 0.01). The increases in PICB concentration resulted in a highly significant linear and quadratic response by 96-h dynamic fermentation parameters (p < 0.01). Our results indicate that 0.2% PICB had the best effect on in-vitro rumen fermentation efficiency and reduced greenhouse gas production.

scholarly journals Design and development of a continuous culture system for studying rumen fermentation

1989 ◽  
Vol 61 (5) ◽  
pp. 463-473
Author(s):  
Harri Miettinen ◽  
Jouko Setälä
Keyword(s):  
Continuous Culture ◽  
Culture System ◽  
Artificial Saliva ◽  
Present Report ◽  
Ammonia Concentration ◽  
Rumen Fermentation ◽  
Liquid Volume ◽  
Desktop Computer ◽  
Continuous Culture System

The present report describes an in vitro continuous culture system to simulate rumen fermentation. The complete assembly consists of six culture vessels (liquid volume 700 ml) fed twice daily with finely ground feed. The artificial saliva enters the vessel continuously, and the effluent leaves it continuously through the overflow port. The intermittent stirring of the fermentor content and the pH regulation are automatically controlled by a desktop computer. Two replicate experiments with ten fermentors given a diet of silage (50 %) and barley (50 %) were made in order to evaluate the system. The results indicate that the system reaches steady-state conditions within three to five days, ammonia concentration being an exeption. It takes for the ammonia concentration approximately 11—14 days to stabilize. The plateau values for the total volatile fatty acid (VFA) concentrations, the molar proportions of individual VFAs, and the ammonia concentrations were found to be within the accepted range in the rumen of animals given similar diets or in other artificial rumen systems. There was a tenfold decrease in the numbers of protozoa in the fermentors during the first four days of incubation. However, the average plateau value for the protozoa numbers (2.5 x 104/ml) is in the same range as in the dual flow continuous culture systems. The efficiency of the microbial N production was higher than that usually observed in vivo or in vitro (45 vs. 30 g/kg organic matter digested). The results indicate that this continuous culture system provides a reasonable estimate of rumen fermentation.

scholarly journals Essential oils effect on rumen fermentation and biohydrogenation under in vitro conditions

2014 ◽  
Vol 59 (No. 10) ◽  
pp. 450-459 ◽  
Author(s):  
M. Gunal ◽  
A. Ishlak ◽  
A.A. AbuGhazaleh ◽  
W. Khattab
Keyword(s):  
Fatty Acids ◽  
Essential Oils ◽  
Dose Level ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Rumen Fermentation ◽  
Buffer Solution ◽  
Cinnamon Oil ◽  
Tea Tree

The effects of adding essential oils (EO) at different levels (125, 250, 500 mg/l) on rumen fermentation and biohydrogenation were examined in a rumen batch culture study. Treatments were: control without EO (CON), control with anise oil (ANO), cedar wood oil (CWO), cinnamon oil (CNO), eucalyptus oil (EUO), and tea tree oil (TEO). Essential oils, each dissolved in 1 ml of ethanol, were added to the culture flask containing 40 ml of buffer solution, 2 ml of reduction solution, 10 ml of rumen fluid, 25 mg of soybean oil, and 0.5 g of the diet. After 24 h of incubation in a water batch at 39&deg;C, three samples were collected from each flask and analyzed for ammonia-N, volatile fatty acids (VFA), and fatty acids (FA). Expect for CNO, the proportions of acetate, propionate, and acetate to propionate ratios were not affected (P &gt; 0.05) by EO addition. Addition of CWO, CNO, and TEO reduced total VFA concentrations (P &lt; 0.05) regardless of dose level. The ammonia-N concentration was greater in cultures incubated with EO regardless of dose level. Compared with the CON, the concentrations of C18:0 and trans C18:1 were reduced (P &lt; 0.05) with EO addition regardless of dose level. Compared with the CON, the concentration of linoleic acid was greater (P &lt; 0.05) when EO were added at 500&nbsp;mg/l. EO tested in this study had no effects on VFA profile but significantly reduced the formation of biohydrogenation products (C18:0 and trans C18:1).

scholarly journals Effects of Geraniol and Camphene on in Vitro Rumen Fermentation and Methane Production

2017 ◽  
Vol 48 (2) ◽  
pp. 63-69
Author(s):  
M. Joch ◽  
V. Kudrna ◽  
B. Hučko
Keyword(s):  
Methane Emission ◽  
Methane Production ◽  
Dry Matter ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Positive Control ◽  
Rumen Fermentation ◽  
Dry Matter Digestibility

AbstractThe objective of this study was to determine the effects of geraniol and camphene at three dosages (300, 600, and 900 mg l-1) on rumen microbial fermentation and methane emission in in vitro batch culture of rumen fluid supplied with a 60 : 40 forage : concentrate substrate (16.2% crude protein, 33.1% neutral detergent fibre). The ionophore antibiotic monensin (8 mg/l) was used as positive control. Compared to control, geraniol significantly (P < 0.05) reduced methane production with increasing doses, with reductions by 10.2, 66.9, and 97.9%. However, total volatile fatty acids (VFA) production and in vitro dry matter digestibility were also reduced (P < 0.05) by all doses of geraniol. Camphene demonstrated weak and unpromising effects on rumen fermentation. Camphene did not decrease (P > 0.05) methane production and slightly decreased (P < 0.05) VFA production. Due to the strong antimethanogenic effect of geraniol a careful selection of dose and combination with other antimethanogenic compounds may be effective in mitigating methane emission from ruminants. However, if a reduction in total VFA production and dry matter digestibility persisted in vivo, geraniol would have a negative effect on animal productivity.

In vitromicrobial growth and rumen fermentation of different substrates as affected by the addition of disodium malate

2005 ◽  
Vol 81 (1) ◽  
pp. 31-38 ◽  
Author(s):  
M. L. Tejido ◽  
M. J. Ranilla ◽  
R. García-Martínez ◽  
M. D. Carro
Keyword(s):  
Dry Matter ◽  
Microbial Growth ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Mean Values ◽  
Batch Cultures ◽  
Production Values ◽  
Micro Organisms ◽  
Microbial N

AbstractThe effects of two concentrations of disodium malate on thein vitrofermentation of three substrates differing in their forage: concentrate ratio (0·8: 0·2, 0·5: 0·5 and 0·2: 0·8; g/g dry matter; low-, medium- and high-concentrate substrates, respectively) by rumen micro-organisms were studied using batch cultures. Rumen contents were collected from four Merino sheep offered lucerne hay ad libitum and supplemented daily with 400 g concentrate. Disodium malate was added to the incubation bottles to achieve final concentrations of 0, 4 and 8 mmol/l malate and15N was used as a microbial marker. Gas production was measured at regular intervals from 0 to 120 h of incubation to study fermentation kinetics. When gas production values were corrected for gas released from added malate, no effects (P> 0·05) of malate were detected for any of the estimated gas production parameters. In 17-h incubations, the final pH and total volatile fatty acid (VFA) production were increased (P< 0·001) by the addition of malate, but no changes (P> 0·05) were detected in the final amounts of ammonia-N and lactate. When net VFA productions were corrected for the amount of VFA produced from malate fermentation itself, adding malate did not affect (P> 0·05) the production of acetate, propionate and total VFA. Malate reduced methane (CH4) production by proportionately 0·058, 0·013 and 0·054 for the low-, medium- and high-concentrate substrates, respectively. Adding malate to batch cultures increased (P< 0·01) rumen microbial growth (mean values of 16·6, 18·3 and 18·4 mg of microbial N for malate at 0, 4 and 8 mmol/l, respectively), but did not affect (P> 0·05) its efficiency of growth (55·5, 56·7 and 54·3 mg of microbial N per g of organic matter apparently fermented for malate at 0, 4 and 8 mmol/l, respectively). There were no interactions (P> 0·05) malate × substrate for any of the measured variables, and no differences (P> 0·05) in pH, CH4production and microbial growth were found between malate at 4 and 8 mmol/l. The results indicate that malate had a beneficial effect on in vitro rumen fermentation of substrates by increasing VFA production and microbial growth, and that only subtle differences in the effects of malate were observed between substrates. Most of the observed effects, however, seem to be due to fermentation of malate itself.

A stringent test for the artificial rumen

1962 ◽  
Vol 13 (2) ◽  
pp. 343 ◽  
Author(s):  
FV Gray ◽  
RA Weller ◽  
AF Pilgrim ◽  
GB Jones
Keyword(s):  
Fatty Acids ◽  
Volatile Fatty Acids ◽  
Artificial Saliva ◽  
Two Systems ◽  
Stringent Test ◽  
Specific Activities

In each of three experiments the acetic, propionic, and butyric acids in the rumen of a sheep were labelled with 14C and a sample of the rumen contents was removed to an artificial rumen so that fermentations of the same substrates could be conducted simultaneously in vivo and in vitro. The in vitro fermentations were carried out in a specially designed artificial rumen of the "permeable" type in which the volume of the contents was kept constant while a continuous supply of artificial saliva was introduced. The relationships between the specific activities of the volatile fatty acids during a period of 2 to 3 hr indicated that the relative rates of formation of the acids were similar in the two systems, although in each experiment the fermentation appeared to proceed more slowly in vitro. It is considered that the procedure constitutes a stringent test for the correct functioning of an artificial rumen.

scholarly journals Influence of tannic acid application on alfalfa hay: in vitro rumen fermentation, serum metabolites and nitrogen balance in sheep

animal ◽  
2008 ◽  
Vol 2 (3) ◽  
pp. 381-390 ◽  
Author(s):  
G. Getachew ◽  
W. Pittroff ◽  
E.J. DePeters ◽  
D.H. Putnam ◽  
A. Dandekar ◽  
...  
Keyword(s):  
Tannic Acid ◽  
Nitrogen Balance ◽  
Rumen Fermentation ◽  
Serum Metabolites ◽  
Alfalfa Hay ◽  
In Vitro Rumen Fermentation

scholarly journals Excretion of purine derivatives by ruminants: recycling of allantoin into the rumen via saliva and its fate in the gut

1990 ◽  
Vol 63 (2) ◽  
pp. 197-205 ◽  
Author(s):  
X. B. Chen ◽  
F. D. DeB. Hovell ◽  
E. R. ØRskov
Keyword(s):  
Fatty Acids ◽  
Uric Acid ◽  
Urinary Excretion ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Purine Derivatives ◽  
Complete Destruction ◽  
Series Of Experiments ◽  
Micro Organisms

The saliva of sheep was shown to contain significant concentrations of uric acid (16 (sd) 4.5) μmol/l) and allantoin (120 (sd 16.4) μmol/l), sufficient to recycle purine derivatives equivalent to about 0.10 of the normal urinary excretion. When allantoin was incubated in vitro in rumen fluid, it was degraded at a rate sufficient to ensure complete destruction of recycled allantoin. In a series of experiments in which allantoin was infused into the rumen of sheep fed normally, or into the rumen or abomasum of sheep and the rumen of cattle completely nourished by intragastric infusion of volatile fatty acids and casein, no additional allantoin was recovered in the urine. These losses were probably due to the degradation of allantoin by micro-organisms associated with the digestive tract. It is concluded that all allantoin and uric acid recycled to the rumen via saliva will be similarly degraded. Therefore, the use of urinary excretion of purine derivatives as an estimator of the rumen microbial biomass available to ruminants will need to be corrected for such losses.

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