Interactions between rumen amylolytic and lactate-utilizing bacteria in growth on starch

The growth and metabolism of the rumen amylolytic bacteria Streptococcus bovis, Butyrivibrio fibrisolvens and Bacteroides ruminicola , growing in pure cultures and co-cultures with the rumen lactilytic bacteria Megasphera elsdenii and Veillonella alcalescens were followed. The interaction of amylolytic bacteria with V. alcalescens represents a simple food chain. The interaction with M. elsdenii is more complex, since there is a simultaneous competition for products of the starch degradation.     

Interactions between rumen amylolytic and lactate-utilizing bacteria in growth on starch

The growth and metabolism of the rumen amylolytic bacteria Streptococcus bovis, Butyrivibrio fibrisolvens and Bacteroides ruminicola, growing in pure cultures and co-cultures with the rumen lactilytic bacteria Megasphaera elsdenii and Veillonella alcalescens were followed. The interaction of amylolytic bacteria with V. alcalescens represents a simple food chain. The interaction with M. elsdenii is more complex, since there is a simultaneous competition for products of the starch degradation.     

Effect of pH on the efficiency of growth by pure cultures of rumen bacteria in continuous culture.

A total of 10 strains of rumen bacteria, Selenomonas ruminantium HD4, Megasphaera elsdenii B159, Butyrivibrio fibrisolvens A38, Streptococcus bovis JB1, Lactobacillus vitulinus GA1, Bacteroides ruminicola B14, B. ruminicola GA33, Ruminococcus albus 7, Ruminococcus flavefaciens C94, and Bacteroides succinogenes S85, were grown in energy-limiteH of the medium reservoir was lowered approximately 0.3 pH units, and the energy source concentration remaining in the culture vessel, optical density, cell mass, and pH were determined. A low pH appeared to have a detrimental effect on cell yields. Large variations were seen among strains in both the magnitude of yield depressions at lower pH values and in the pH at which the culture washed out. Lactate analysis indicated ta are discussed in relation to the effect of pH on the efficiency of protein synthesis in the rumen and rumen microbial ecology.     

Comparative growth rates of various rumen bacteria in clarified rumen fluid from cows and sheep fed different diets.

Pure cultures of strains of different species of rumen bacteria were grown in filter-sterilized rumen fluid supplemented with glucose, bicarbonate, and reducing agent (cysteine and sulfide). Growth rates were determined in a series of experiments. Strains of species most abundant in the rumen grew more rapidly than strains of less abundant bacteria. Ammonia, amino acids, and peptides increased growth rates to some extent, but the greatest stimulatory effect for less abundant bacteria was provided by other factors, present in yeast extract. Factors released from lysates of mixed rumen microbes stimulated growth, but their rate of release was slow. It was concluded that, besides energy and nitrogen sources, growth factors of an as-yet-undetermined nature probably play an important role in determining the predominance of different bacterial species in the rumen.     

Comparative growth rates of various rumen bacteria in clarified rumen fluid from cows and sheep fed different diets.

Pure cultures of strains of different species of rumen bacteria were grown in filter-sterilized rumen fluid supplemented with glucose, bicarbonate, and reducing agent (cysteine and sulfide). Growth rates were determined in a series of experiments. Strains of species most abundant in the rumen grew more rapidly than strains of less abundant bacteria. Ammonia, amino acids, and peptides increased growth rates to some extent, but the greatest stimulatory effect for less abundant bacteria was provided by other factors, present in yeast extract. Factors released from lysates of mixed rumen microbes stimulated growth, but their rate of release was slow. It was concluded that, besides energy and nitrogen sources, growth factors of an as-yet-undetermined nature probably play an important role in determining the predominance of different bacterial species in the rumen.     

Effect of fatty acids on growth of conjugated-linoleic-acids-producing bacteria in rumen

Microorganisms with high activity of linoleic acid delta12-cis,delta11-trans-isomerase were isolated from the digestive tract of ruminants and characterized. The isolate with the highest isomerase activity was identified as Pseudobutyrivibrio ruminis. The susceptibility of this strain to 3 fatty acids added to the grow medium was determined. A significant inhibition of bacterial growth (during a 3-d period) by linoleic acid (0.1 %) and oleic acid (5 ppm) was observed; no inhibition was found in the presence of stearic acid.     

Effect of titanium (III) citrate as reducing agent on growth of rumen bacteria.

We compared the growth of 10 strains of rumen bacteria in an anaerobic medium reduced with cysteine hydrochloride, dithiothreitol, or titanium (III) citrate. The redox potential of medium reduced with cysteine hydrochloride was -167.8 mV; with dithiothreitol it was -175.8 mV; and with titanium(III) citrate it was -302.4 mV at a concentration of 5 X 10(-4) M titanium and -403.9 mV at 2 X 10(-3) M titanium. Maximum growth of the strains was generally lower with dithiothreitol or titanium(III) citrate than with cysteine hydrochloride, although growth was greater than in medium lacking an added reducing agent. Strains for which cysteine was required or markedly stimulatory grew only poorly with titanium(III) citrate. No strain grew in medium with sodium citrate as the energy source. Titanium(III) citrate could be used to reduce anaerobic media for some rumen bacteria if the exclusion of a sulfur-containing reducing agent is required.     

Stoichiometry of glucose and starch splitting by strains of amylolytic bacteria from the rumen and anaerobic digester

The stoichiometry of glucose and starch splitting by the amylolytic bacteria Streptococcus bovis, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Eubacterium ruminantium and Clostridium sp. was followed. There were many differences in the ratios of metabolites and in growth yields, as well as in the cell composition, between the growth on glucose and starch. The bacteria employ different nutritional strategies with respect to both energy sources.     

Stoichiometry of glucose and starch splitting by strains of amylolytic bacteria from the rumen and anaerobic digester

The stoichiometry of glucose and starch splitting by the amylolytic bacteria Streptococcus bovis, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Eubacterium ruminantium and Clostridium sp. was followed. There were many differences in the ratios of metabolites and in growth yields, as well as in the cell composition, between the growth on glucose and starch. The bacteria employ different nutritional strategies with respect to both energy sources.     

The effect of pH and 1,4-dithiothreitol on the adhesion of rumen bacteria

In experiments with ram rumen fluid the pH optimum for adhesion of rumen bacteria to cellulose and starch was found to be near 6. Addition of 1,4-dithiothreitol increased the adhesion logarithmically.     

Effect of whole-plant corn silage treated with lignocellulose-degrading bacteria on growth performance,rumen fermentation,and rumen microflora in sheep

Lignification of cellulose limits the effective utilisation of fibre in plant cell wall. Lignocellulose-degrading bacteria secrete enzymes that decompose lignin and have the potential to improve fibre digestibility. Therefore, this study aimed to investigate the effect of whole-plant corn silage inoculated with lignocellulose-degrading bacteria on the growth performance, rumen fermentation, and rumen microbiome in sheep. Twelve 2-month-old male hybrid sheep (Dorper ♂ × small-tailed Han ♀) were randomly assigned into two dietary groups (n = 6): (1) untreated whole-plant corn silage (WPCS) and (2) WPCS inoculated with bacterial inoculant (WPCSB). Whole-plant corn silage inoculated with bacterial inoculant had higher in situ NDF digestibility than WPCS. Sheep in the WPCSB group had significantly higher average daily gain, DM intake, and feed conversion rate than those in the WPCS group (P < 0.05). Furthermore, higher volatile fatty acid concentrations were detected in WPCSB rumen samples, leading to lower ruminal pH (P < 0.05). The WPCSB group showed higher abundance of Bacteroidetes and lower abundance of Firmicutes in the rumen microbiome than the WPCS group (P < 0.05). Multiple differential genera were identified, with Prevotella being the most dominant genus and more abundant in WPCSB samples. Moreover, the enriched functional attributes, including those associated with glycolysis/gluconeogenesis and citrate cycle, were more actively expressed in the WPCSB samples than in the WPCS samples. Additionally, certain glucoside hydrolases that hydrolyse the side chains of hemicelluloses and pectins were also actively expressed in the WPCSB microbiome. These findings suggested that WPCSB increased NDF digestibility in three ways: (1) by increasing the relative abundance of the most abundant genera, (2) by recruiting more functional features involved in glycolysis/gluconeogenesis and citrate cycle pathways, and (3) by increasing the relative abundance and/or expression activity of the glucoside hydrolases involved in hemicellulose and pectin metabolism. Our findings provide novel insights into the microbial mechanisms underlying improvement in the growth performance of sheep/ruminants. However, the biological mechanisms cannot be fully elucidated using only metagenomics tools; therefore, a combined multi-omics approach will be used in subsequent studies.     

Effect of alfalfa fiber substrate on culture counts of rumen bacteria.

A medium has been developed using alfalfa fiber as the sole substrate. It gave high culture counts (3 X 10(9) to 8 X 10(9)/ml) of rumen bacteria. When this medium was combined with the medium 98-5 of Bryant and Robinson, modified to contain 33% rumen fluid instead of 40% clarified rumen fluid, a higher count was obtained than with either medium alone.     

Some effects of uncouplers and inhibitors on growth and electron transport in rumen bacteria.

Uncouplers and inhibitors of electron transport affected growth and electron transport of rumen bacteria in various ways. Selenomonas ruminantium was not affected by inhibitor and uncoupler concentrations which affected growth and electron transport of Bacteroides ruminicola, B. succinogenes, and Butyrivibrio fibrisolvens. Inhibitors, when active, led to accumulation of reduced electron carriers before the site of action, but differences were found among organisms in the site of action of these inhibitors. Uncouplers reduced the glucose molar growth yields (Ygluc) of B. ruminicola, B. succinogenes, and B. fibrisolvens compared with those obtained without uncouplers. The extent of Ygluc reduction accompanying inhibitor exposure reflected electron transport chain structure. S. ruminantium appeared to obtain its adenosine 5'-triphosphate from substrate-level processes only. The other organisms studied appeared to obtain adenosine 5'-triphosphate both from substrate-level processes and from electron transport but differed in the amount of adenosine 5'-triphosphate obtained from glucose catabolism and in the proportions of adenosine 5'-triphosphate obtained from substrate-level reactions and electron transport.     

Competitive ability of amylolytic bacteria in activated sludge.

Shifts were induced into the microbial community of activated sludge by the pulse addition of soluble starch. The subsequent changes of amylolytic and proteolytic microbial populations were recorded. Four amylolytic strains were isolated and characterized with regard to carrying capacity, specific surface and growth kinetics. The competitive ability of these strains was studied by means of two-member competition experiments. These experiments were analysed according to the Lotka-Volterra model and the de Wit method. The different results obtained suggest that the dominance of the amylolytic Pseudomonas sp. (code 01) is based on a combined occurrence of high amylolytic activity, large relative cell surface, high maximum specific growth rate and reduced sensitivity towards associated proteolytic populations.     

Some effects of ethyl alcohol on the growth of rumen bacteria

The effects of ethyl alcohol on the growth rate and yield of representative isolates of obligately anaerobic rumen bacteria were studied. Yield was monitored as turbidity at 600 nm until stationary phase was obtained and quantitated as protein per milliter. Rate was determined as the fastest rate of turbidity doubling. With very few exceptions, within the concentration range allowing growth, no effects of alcohol concentration were observed on rate, but substantial effects were found on yield. Growth of most of the organisms studied was completely inhibited at an alcohol concentration between 3% and 7% vol/vol, but the growth ofBacteroides amylophilus andRuminococcus flavefaciens was drastically reduced or eliminated at a 1% alcohol concentration. It is likely that major physiological and biochemical differences exist among anaerobic rumen bacteria, and taxonomically similar organisms, as a function of alcohol exposure.