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1.
The cell number of Selenomonas ruminantium (S. ruminantium) that reduces nitrate and nitrite in the rumen was usually 8–10% of the total number of S. ruminantium (an order of 106/mL). The percentage was not affected by the roughage/concentrate ratio or nitrate content of the diet in 2 weeks. However, feeding a high‐nitrate diet for 12 weeks increased the percentage. The percentage of lactate‐using S. ruminantium, such as the ssp. lactilytica, was less than 1% of the total number of S. ruminantium. No S. ruminantium was found that used formate as an electron donor for nitrate and nitrite reduction. Lactate and H2 appeared to be important for nitrate and nitrite reduction by S. ruminantium. Nitrate reduction by S. ruminantium was enhanced by the coexistence of amylolytic bacteria in a medium containing starch, and as a result, nitrite accumulation increased. Coexistence of cellulolytic bacteria facilitated the growth of S. ruminantium in a medium containing cellulose, and consequently increased nitrite reduction. In order to suppress nitrite accumulation in the rumen, it may be important to enhance fiber digestion.  相似文献   

2.
This study investigated the effects of dietary nitrate addition on ruminal fermentation characteristics and microbial populations in goats. The involvement of Selenomonas ruminantium in nitrate and nitrite reduction in the rumen was also examined. As the result of nitrate feeding, the total concentration of ruminal volatile fatty acids decreased, whereas the acetate : propionate ratio and the concentrations of ammonia and lactate increased. Populations of methanogens, protozoa and fungi, as estimated by real‐time PCR, were greatly decreased as a result of nitrate inclusion in the diet. There was modest or little impact of nitrate on the populations of prevailing species or genus of bacteria in the rumen, whereas Streptococcus bovis and S. ruminantium significantly increased. Both the activities of nitrate reductase (NaR) and nitrite reductase (NiR) per total mass of ruminal bacteria were increased by nitrate feeding. Quantification of the genes encoding NaR and NiR by real‐time PCR with primers specific for S. ruminantium showed that these genes were increased by feeding nitrate, suggesting that the growth of nitrate‐ and nitrite‐reducing S. ruminantium is stimulated by nitrate addition. Thus, S. ruminantium is likely to play a major role in nitrate and nitrite reduction in the rumen.  相似文献   

3.
The effect of ethanol on nitrate and nitrite reduction was examined by conducting in vitro experiments with mixed ruminal microbes. The addition of ethanol to cultures of mixed ruminal microbes stimulated nitrate reduction, and, to a greater extent, nitrite reduction, which resulted in a decrease in nitrite accumulation. However, known nitrate‐reducing ruminal bacteria, such as Selenomonas ruminantium, Veillonella parvula and Wolinella succinogenes, were unable to utilize ethanol directly as an electron donor for nitrate reduction. No nitrate‐reducing bacterium capable of utilizing ethanol was found in the rumen of goats. However, when mixed ethanol‐utilizing, hydrogen gas (H2)‐producing bacteria (Ruminococcus albus and Ruminococcus flavefaciens) were added to the culture of the mixed nitrate‐reducing bacteria described above, nitrate and nitrite reduction was observed. These results suggest that the nitrate‐reducing bacteria utilized the H2 that was produced from ethanol oxidation by the ethanol‐utilizing bacteria as an electron donor. It is conceivable that the stimulation of nitrate and nitrite reduction by ethanol, observed in the culture of mixed ruminal microbes, was a result of electron transfer from ethanol to nitrate, and nitrite through H2, that is, ‘interspecies hydrogen transfer’ from ethanol‐metabolizing bacteria to nitrate‐reducing bacteria. Thus, the addition of ethanol to high‐nitrate diets may be effective for preventing nitrate poisoning. Furthermore, methane production was reduced to less than one‐third by the addition of mixed nitrate‐reducing bacteria to the co‐culture of mixed methanogens with mixed ethanol‐utilizing bacteria incubated in a medium containing ethanol and nitrate. Therefore, the addition of ethanol and nitrate may decrease methanogenesis without suppressing overall fermentation in the rumen.  相似文献   

4.
In this study, we examined the potential of Megasphaera elsdenii as a probiotic agent in combination with sodium gluconate. We also examined the combination of M. elsdenii with probiotic lactic acid bacteria (LAB), Lactobacillus acidophilus, to enhance butyrate production in the large intestine using a pig cecal in vitro model under the presence of sodium gluconate. Compared to the uninoculated control culture (with only diluted cecal digesta), the addition of M. elsdenii and LAB to the culture stimulated significantly the production of n‐butyrate (60% increase) and n‐valerate (50% increase) after 24 h incubation. n‐Butyrate is regarded as the most important short‐chain fatty acid in the large intestine because it stimulates the proliferation of epithelial cells, mucus release and water and mineral absorption. Therefore, lactate‐utilizing butyrate producers such as M. elsdenii play a significant role in supporting the health‐beneficial effects of lactogenic prebiotics and probiotics LAB. The potential of M. elsdenii as a probiotic was also suggested.  相似文献   

5.
Methyl‐mannosylerythritol lipid (MEL), a new sugar esterified lipid synthesized by Pseudozyma aphidis, was assessed for its functionality in modulating rumen fermentation and microbiota toward more propionate and less methane production. A pure culture study using rumen representatives showed that MEL selectively inhibited the growth of most Gram‐positive bacteria including Streptococcus bovis, ruminococci, and Fibrobacter succinogenes, but not Gram‐negative bacteria such as Megasphaera elsdenii, Succinivibrio dextrinosolvens, and Selenomonas ruminantium. A batch culture study revealed that MEL significantly decreased methane production in a dose‐dependent manner with accumulation of hydrogen, while propionate production was enhanced. A continuous culture (Rusitec) study confirmed all of these changes. A feeding study revealed that sheep fed a MEL diet showed an increased proportion of propionate, while proportions of acetate and butyrate were decreased without affecting total VFA level. These changes disappeared after cessation of MEL feeding. Based on these results, dietary application of MEL can favorably modify rumen fermentation in terms of the efficiency of dietary energy utilization.  相似文献   

6.
The regulation of lactic acid production, the regulation of lactate fermentation and the role of lactate as intermediate in the rumen metabolism was studied.The pH had a pronounced effect on all three processes and therefore buffer capacity of the rumen contents is also described.Starch gave much less rise to lactic acidosis than soluble sugars, as glucose and fructose. Most bacteria grow faster and therefore produce more lactic acid when amino acids and/or soluble proteins are present in the diet.Activity of LDH (lactate dehydrogenase) of mixed rumen microorganisms is regulated by the NADH/NAD (H) balance and the ATP concentration. About 60% of the LDH in mixed rumen microorganisms is fructose-1, 6-diphosphate independent. Megasphaera elsdenii ferments 60 to 80% of the lactate fermented in the rumen of dairy cattle.Lactate accumulates only when the glycolytic flux (hexose units fermented per unit time per microorganism) is high. During adaptation, the glycolytic flux is increased and lactate may accumulate. After adaptation to a certain diet, the number of microorganisms is changed and the glycolytic flux again is normal and lactate is only a minor intermediate in rumen metabolism.  相似文献   

7.
Two consecutive experiments were carried out to determine efficacy of Megasphaera elsdenii inoculation in alleviation of subacute ruminal acidosis (SARA). In the first experiment, SARA was induced by feeding corn‐ and wheat‐based diets (20%, 40%, 60% and 80% of TMR, DM basis) in six ruminally cannulated heifers. Continuous pH was obtained using data loggers embedded in rumen. In corn (80%)‐ and wheat (60%)‐based diets ruminal pH ranged from 5.2 to 5.6 for 7.77 and 5.93 hr. In the second experiment (5 day), M. elsdenii (200 ml; 2.4 x 1010 cfu/ml) was inoculated during the first two days. During the SARA induction period, M. elsdenii and S. bovis in rumen liquor were more abundant in wheat‐based feeding (7.97 and 8.77) than in corn‐based feeding (7.06 and 7.95 per ml, log basis; p < 0.0001 for both). M. elsdenii inoculation increased total volatile fatty acids (VFA) concentration when corn‐based diet was fed, whereas it decreased total VFA concentration when wheat‐based diet was fed (p < 0.004). There was a decrease in the propionic acid proportion (24.04%–19.08%; p < 0.002), whereas no alteration in lactate and ammonia concentrations was observed. M. elsdenii inoculation increased protozoa count (from 5.39 to 5.55 per ml, log basis; p < 0.009) and decreased S. bovis count (from 9.18 to 7.95 per ml, log basis; p < 0.0001). The results suggest that M. elsdenii inoculation may help prevent SARA depending on dietary grain through altering rumen flora as reflected by a decrease in S. bovis count and an increase in protozoa count.  相似文献   

8.
To investigate the rumen bacterial interaction between cellulolytic Ruminococcus flavefaciens and non‐cellulolytic Selenomonas ruminantium, fiber digestibility and fermentation products were determined in defined cultures consisting of these two species. Avicel, orchardgrass hay, rice straw and alfalfa hay were used as substrates for 72 h incubation to monitor digestibility, volatile fatty acids, succinate, lactate and bacterial number. In monoculture, R. flavefaciens digested the fiber sources at 21–32%, while S. ruminantium strains did not. When R. flavefaciens was cocultured with one of three different strains (GA192, S137 and S150) of S. ruminantium, fiber digestion exceeded the value recorded by R. flavefaciens alone. In particular, cocultures with S. ruminantium S137 showed significantly higher digestibility for all the fiber sources than R. flavefaciens alone (P < 0.05). Propionate production and growth of S. ruminantium was notable in all cocultures but not in monocultures. Succinate was accumulated in monoculture of R. flavefaciens, while the accumulation was not observed in cocultures. These results indicate that R. flavefaciens provides fiber hydrolysis products to S. ruminantium as growth substrates. In addition, S. ruminantium could activate R. flavefaciens by rapidly consuming the products. Such cross‐feeding between cellulolytic and non‐cellulolytic bacteria could enhance fiber digestion, although the extent of the enhancement may depend on strain combinations.  相似文献   

9.
An in vitro study was conducted to quantitatively investigate the metabolism of pipecolic acid (Pip), a neuromodulator, by mixed rumen bacteria (B), mixed rumen protozoa (P), a combination of B and P (BP), species‐enriched rumen protozoal suspension (Polyplastron sp., Diploplastron sp., entodinia and Entodinium caudatum) and pure cultures of several isolates of rumen bacteria (Prevetolla bryantii, Prevetolla albensis, Streptococcus bovis, Veillonella parvula, Megasphaera elsdenii and Ruminococcus albus). Only P produced Pip from L‐lysine (1.0 mmol/L L‐Lys) at a rate of 83.5 ± 1.6 µmol/L/h and even in BP, Pip was produced from L‐Lys by P and increased at a rate of 31.2 ± 3.8 µmol/L/h. Pip production by P was highest when the substrate (L‐Lys) concentration was 6 mmol/L and then the rate was 580 ± 36 µmol/L/h. Pipecolic acid production by P suspension enriched with different species of protozoa showed that Polyplastron sp. had the highest Pip production rate of 0.907 ± 0.092 µmol/L/mg protozoal protein per h, and Diploplastron sp. had the lowest rate of 0.55 ± 0.13 µmol/L/mg protozoal protein per h. The addition of D‐Lys (1.0 mmol/L) as a substrate to the P suspension revealed that P were also able to produce Pip from D‐Lys, though at a lower rate (1/3) compared with L‐Lys (1.0 mmol/L), suggesting the presence of epimerases in P. It was confirmed that B were unable to produce Pip from L‐ or D‐Lys. Only B degraded Pip (1.0 mmol/L) after a lag phase at a rate of 56.0 ± 1.5 µmol/L/h. The B suspension was able to degrade D‐Lys, though the products were not identified. Pip degradation by pure culture of some species of rumen bacteria showed that P. bryantii and R. albus had the highest rate followed by P. albensis, S. bovis and M. elsdenii with a low rate of Pip degradation. Veillonella parvula showed no ability to degrade Pip. The results suggest that a fairly large proportion of rumen‐produced Pip is likely to be absorbed by the host animal before degradation by rumen bacteria.  相似文献   

10.
The fibrolytic rumen bacterium Ruminococcus flavefaciensOS14 was isolated from swamp buffalo and its phylogenetic, ecological and digestive properties were partially characterized. Isolates from rumen contents of four swamp buffalo were screened for fibrolytic bacteria; one of the 40 isolates showed a distinctive feature of solubilizing cellulose powder in liquid culture and was identified as R. flavefaciens based on its 16S ribosomal DNA sequence. This isolate, OS14, was employed for detection and digestion studies, for which a quantitative PCR assay was developed and defined cultures were tested with representative forages in Thailand. OS14 was phylogenetically distant from other isolated and uncultured R. flavefaciens and showed limited distribution among Thai ruminants but was absent in Japanese cattle. OS14 digested rice straw and other tropical forage to a greater extent than the type strain C94 of R. flavefaciens. OS14 produced more lactate than C94, and digested para grass to produce propionate more extensively in co‐culture with lactate‐utilizing Selenomonas ruminantium S137 than a co‐culture of C94 with S137. These results indicate that phylogenetically distinct OS14 could digest Thai local forage more efficiently than the type strain, possibly forming a symbiotic cross‐feeding relationship with lactate‐utilizing bacteria. This strain might be useful for future animal and other industrial applications.  相似文献   

11.
Weaning causes atrophy of intestinal mucosa and a drop of IgA protection in piglets which increases vulnerability to pathogenic infections. Probiotic lactobacilli may support recovery from such weaning stresses. Butyrate‐produce bacteria may support the growth of colonic mucosa. Megasphaera elsdenii, a lactate‐utilizing butyrate producer, may help butyrate production particularly when combined with lactobacilli. Weaned piglets (Experiment 1: 20 days old, Experiment 2: 28 days old) were orally dosed once a day with either (L) 1010 (cell/dose) L. plantarum Lq80, or (LM) 1010 (cell/dose) Lq80 with 109 (cell/dose) M. elsdenii iNP‐001. Lq80 was contained in capsules resistant to gastric digestion. M. elsdenii was contained in capsules resistant to gastric and intestinal digestion. An untreated control (C) was also prepared. After 2 weeks of administration, L. plantarum enhanced the recovery from the villous atrophy in both experiments. The rectal and colonic IgA tended to be higher in L and LM than in C in Experiment 1. Colonic butyrate was higher in LM than in the others in Experiment 1. The thickness of the colonic mucosa was greater in LM than in the others in Experiment 1. In early weaned piglets, the effects of L. plantarum and M. elsdenii were clear.  相似文献   

12.
Thirty calves were randomly assigned to two treatments and fed until weaning [42 days (d) of age]. Treatments were a control group (n = 15), which did not receive Megasphaera elsdenii (Me0) and a M. elsdenii group, which received a 50‐ml oral dose of M. elsdenii NCIMB 41125 (108 CFU/ml) at day 14 day of age (Me14). Calves were given colostrum for the first 3 day followed by limited whole milk feeding. A commercial calf starter was offered ad libitum starting at day 4 until the end of the study. Fresh water was available throughout the study. Feed intake and growth were measured. Blood samples were collected via jugular venipuncture to determine β‐hydroxybutyrate (BHBA) concentrations. Fourteen male calves (seven per group) were euthanised on day 42 and digestive tracts harvested. Reticulo‐rumen weight was determined and rumen tissue samples collected from the cranial and caudal sacs of the ventral and dorsal portions of the rumen for measurements of papillae length, papillae width and rumen wall thickness. Dosing with M. elsdenii NCIMB 41125 improved starter dry matter intake (DMI), weaning body weight (BW) and tended to improve average daily gain. Calves in Me14 group had greater plasma BHBA concentration than Me0‐calves during the last 3 weeks of the trial and had at day 42 greater reticulo‐rumen weight, papillae width and papillae density compared to Me0. No differences in rumen wall thickness or papillae length were observed between the two groups. Total volatile fatty acids, acetate and propionate production did not differ between treatments, but butyrate production was greater in Me14 than Me0. Dosing M. elsdenii NCIMB 41125 showed benefit for calves with improved feed intake and rumen development suggesting increased epithelium metabolism and improved absorption of digestive end products.  相似文献   

13.
The aim of this study was to examine the effects of the acid-tolerant engineered bacterial strain Megasphaera elsdenii H6F32 (M. elsdenii H6F32) on ruminal pH and the lactic acid concentrations in simulated rumen acidosis conditions in vitro. A mixed culture of ruminal bacteria, buffer, and primarily degradable substrates was inoculated with equal numbers of M. elsdenii H6 or M. elsdenii H6F32. The pH and lactic acid concentrations in the mixed culture were determined at 0, 2, 4, 6, 8, 10, 12, 14, 16, and 18 h of incubation. Acid-tolerant M. elsdenii H6F32 reduced the accumulation of lactic acid and increased the pH value. These results indicate that acid-tolerant M. elsdenii H6F32 could be a potential candidate for preventing rumen acidosis.  相似文献   

14.
15.
Microalgae might be considered as an alternative source of fat and/or protein for ruminant's diets. However, changes in populations of ruminal micro‐organisms associated with biohydrogenation process, methane and ammonia production in response to microalgae dietary supplementation have not been well characterized. Thus, 16 cross‐bred goats were divided into two groups. Each goat of both groups was fed individually with alfalfa hay and concentrates separately. The concentrates of the control group had no microalgae while those of the treated group were supplemented with 10 g lyophilized Chlorella vulgaris/kg concentrate (chlor). On the 30th experimental day, samples of rumen fluid were collected for microbial DNA extraction, fatty acid profile and enzyme activity analyses. The results showed that the chlor diet compared with the control increased significantly the populations of Methanosphaera stadtmanae, Methanobrevibacter ruminantium and Methanogens bacteria and protozoa in the rumen of goats. A significant reduction in the cellulase activity and in the abundance of Ruminococcus albus, and a significant increase in the protease activity and in the abundance of Clostridium sticklandii in the rumen liquid of goats fed with the chlor diet, compared with the control, were found. Chlorella vulgaris supplementation promoted the formation of trans C18:1, trans‐11 C18:1 and monounsaturated fatty acids (MUFA), while the proportions of C18:0 and long‐chain fatty acids (LCFA) reduced significantly in the rumen liquid of goats. This shift in ruminal biohydrogenation pathway was accompanied by a significant increase in Butyrivibrio fibrisolvens trans C18:1‐producing bacteria. In conclusion, the supplementation of diets with microalgae needs further investigation because it enhances the populations of methane‐producing bacteria and protozoa.  相似文献   

16.
Real‐time polymerase chain reaction (PCR) assays for 11 representative rumen bacterial species were validated. The sensitivity was tested by using the serially diluted target 16S rDNA from respective bacterial species. The recovery of the target DNA and the assay reproducibility were determined using DNA from rumen fluid spiked with different quantities of the target. Minimum detection levels for the target were 10–100 copies in pure culture. The recovery of the added target ranged from 82.4 to 116.6%. The intra‐ and inter‐assay variations of each assay were <9.4 and <12.6%, respectively. Therefore, the real‐time PCR assays evaluated in the present study are considered to be sufficiently reliable for monitoring all 11 bacterial species in the rumen. The assays were then applied to the monitoring of the bacterial species attached to ruminally incubated rice straw. Among the monitored fibrolytic species, Fibrobacter succinogenes was found to be the most dominant, accounting for 2.61% of total bacteria after 24 h incubation. Selenomonas ruminantium and Streptococcus bovis, non‐fibrolytics, were detected on the rice straw at 8.96% and 1.16% of total bacteria, respectively. Such high levels of non‐fibrolytics on the plant fiber suggest a synergistic relationship between fibrolytics and non‐fibrolytics.  相似文献   

17.
In Plasmodium berghei, sephadex G‐200 purified lactate dehydrogenase (LDH) fraction immunized mice did not exhibit protection when challenged with 1 × 106P. berghei‐parasitized erythrocytes. However, LDH immunized mice seroconverted and showed an antibody titre of 1:2048 by indirect haemagglutination (IHA) and 1:160 by indirect fluorescent antibody (IFA) assays. Fluorescence was distributed evenly on P. berghei‐parasitized red cells showing no specific location of parasite LDH. Anti‐LDH antibodies supplemented in 19 h in vitro culture of P. berghei exhibited 9.2% invasion inhibition into the fresh red cells.  相似文献   

18.
The effects of non‐starch‐polysaccharide‐degrading enzymes, added to a maize silage‐ and grass silage‐based total mixed ration (TMR) at least 14 h before feeding, on the rumen bacterial population were investigated. Six non‐lactating Holstein Friesian cows were allocated to three treatment groups using a duplicate 3 × 3 Latin square design with three 31‐day periods (29 days of adaptation and 2 days of sampling). Treatments were control TMR [69% forage and 31% concentrates on a dry matter (DM) basis] or TMR with 13.8 or 27.7 ml/kg of feed DM of Roxazyme G2 liquid with activities (U/ml enzyme preparation) of xylanase 260 000, β‐glucanase 180 000 and cellulase 8000 (DSM Nutritional Products, Basel, Switzerland). The concentrations of 16S rDNA of Anaerovibrio lipolytica, Fibrobacter succinogenes, Prevotella ruminicola, Ruminococcus flavefaciens, Selenomonas ruminantium and Treponema bryantii, and their relative percentage of total bacteria in rumen samples obtained before feeding and 3 and 7 h after feeding and from two rumen fractions were determined using real‐time PCR. Sampling time had only little influence, but bacterial numbers and the composition of the population differed between the transition layer between rumen fluid and the fibre mat (fraction A) and the rumen fluid (fraction B) highlighting the importance to standardize sampling. The 16S rDNA copies of total bacteria and the six bacterial species as well as the population composition were mainly unaffected by the high levels of exogenous enzymes supplemented at all sampling times and in both rumen fractions. Occasionally, the percentages of the non‐fibrolytic species P. ruminicola and A. lipolytica changed in response to enzyme supplementation. Some increases in the potential degradability of the diet and decreases in lag time which occurred collaterally indicate that other factors than changes in numbers of non‐particle‐associated bacteria are mainly responsible for the effects of exogenous enzymes.  相似文献   

19.
Colonization patterns of representative rumen bacteria were compared between untreated rice straw (UTS) and sodium hydroxide‐treated rice straw (SHTS). UTS and SHTS were incubated in the rumen of sheep for 10 min, 1, 2, 6, 12, 24, 48 and 96 h using the nylon bag method. The population sizes of 13 representative bacterial species or groups were quantified by real‐time PCR. The total bacterial population size (abundance) was similar in both UTS and SHTS. Fibrobacter succinogenes showed a higher population size compared to other fibrolytic species and was detected at a higher level in SHTS (3.7%) than in UTS (2.6%). Ruminococcus albus and Ruminococcus flavefaciens were also detected at higher levels in SHTS (0.15% and 0.29%) than in UTS (0.03% and 0.18%). Population sizes of non‐fibrolytic species, such as Selenomonas ruminantium, Anaerovibrio lipolytica and Succinivibrio dextrinosolvens were higher in UTS than in SHTS. Coefficient of determination (r2) on population changes between bacterial species or groups were higher in UTS than in SHTS, suggesting the necessity of stronger bacterial interactions for UTS digestion. Therefore, not only colonization of fibrolytic species, but also synergistic interactions between different bacterial species may be key to the ruminal digestion of rice straw.  相似文献   

20.
The aim of our experiment was to study how synbiotics are able to deal with the problems of post‐weaning piglets. Lactobacillus plantarum – BiocenolTM LP96 (CCM 7512), Lactobacillus fermentum – BiocenolTM LF99 (CCM 7514) and flaxseed (rich in n‐3 polyunsaturated fatty acids) were administered to 36 conventional piglets from a problematic breed with confirmed presence of enterotoxigenic Escherichia coli and Coronavirus. The experimental piglets were supplied with probiotic cheeses and crushed flax‐seed in the period starting 10 days before weaning and lasting up to 14 days post‐weaning. Piglets in the control group were supplied only control cheese. The impact of such additives on the release of lactate dehydrogenase (LDH; spectroscopic and electrophoretic assay), alteration of immunity (index of metabolic activity), jejunum histology (light microscopy), and health of conventional piglets from a problematic breed (monitoring of hematology, consistency and moisture of feces and body temperature) were examined. We found significant decrease in LDH leakage in the blood serum and tissue extracts, indicating better cell membrane integrity in the individual organs of animals. Probiotics and flaxseed applied together seem to be a good source of nutrients to improve the immune status and the integrity of jejunum mucosa during infection. © 2015 Japanese Society of Animal Science  相似文献   

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