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Sample records for rumen microbial protein

  1. Maximizing efficiency of rumen microbial protein production

    PubMed Central

    Hackmann, Timothy J.; Firkins, Jeffrey L.

    2015-01-01

    Rumen microbes produce cellular protein inefficiently partly because they do not direct all ATP toward growth. They direct some ATP toward maintenance functions, as long-recognized, but they also direct ATP toward reserve carbohydrate synthesis and energy spilling (futile cycles that dissipate heat). Rumen microbes expend ATP by vacillating between (1) accumulation of reserve carbohydrate after feeding (during carbohydrate excess) and (2) mobilization of that carbohydrate thereafter (during carbohydrate limitation). Protozoa account for most accumulation of reserve carbohydrate, and in competition experiments, protozoa accumulated nearly 35-fold more reserve carbohydrate than bacteria. Some pure cultures of bacteria spill energy, but only recently have mixed rumen communities been recognized as capable of the same. When these communities were dosed glucose in vitro, energy spilling could account for nearly 40% of heat production. We suspect that cycling of glycogen (a major reserve carbohydrate) is a major mechanism of spilling; such cycling has already been observed in single-species cultures of protozoa and bacteria. Interconversions of short-chain fatty acids (SCFA) may also expend ATP and depress efficiency of microbial protein production. These interconversions may involve extensive cycling of intermediates, such as cycling of acetate during butyrate production in certain butyrivibrios. We speculate this cycling may expend ATP directly or indirectly. By further quantifying the impact of reserve carbohydrate accumulation, energy spilling, and SCFA interconversions on growth efficiency, we can improve prediction of microbial protein production and guide efforts to improve efficiency of microbial protein production in the rumen. PMID:26029197

  2. Effects of synchronicity of carbohydrate and protein degradation on rumen fermentation characteristics and microbial protein synthesis.

    PubMed

    Seo, J K; Kim, M H; Yang, J Y; Kim, H J; Lee, C H; Kim, K H; Ha, Jong K

    2013-03-01

    A series of in vitro studies were carried out to determine i) the effects of enzyme and formaldehyde treatment on the degradation characteristics of carbohydrate and protein sources and on the synchronicity of these processes, and ii) the effects of synchronizing carbohydrate and protein supply on rumen fermentation and microbial protein synthesis (MPS) in in vitro experiments. Untreated corn (C) and enzyme-treated corn (EC) were combined with soy bean meal with (ES) and without (S) enzyme treatment or formaldehyde treatment (FS). Six experimental feeds (CS, CES, CFS, ECS, ECES and ECFS) with different synchrony indices were prepared. Highly synchronous diets had the greatest dry matter (DM) digestibility when untreated corn was used. However, the degree of synchronicity did not influence DM digestibility when EC was mixed with various soybean meals. At time points of 12 h and 24 h of incubation, EC-containing diets showed lower ammonia-N concentrations than those of C-containing diets, irrespective of the degree of synchronicity, indicating that more efficient utilization of ammonia-N for MPS was achieved by ruminal microorganisms when EC was offered as a carbohydrate source. Within C-containing treatments, the purine base concentration increased as the diets were more synchronized. This effect was not observed when EC was offered. There were significant effects on VFA concentration of both C and S treatments and their interactions. Similar to purine concentrations, total VFA production and individual VFA concentration in the groups containing EC as an energy source was higher than those of other groups (CS, CES and CFS). The results of the present study suggested that the availability of energy or the protein source are the most limiting factors for rumen fermentation and MPS, rather than the degree of synchronicity.

  3. Effects of grain source, grain processing, and protein degradability on rumen kinetics and microbial protein synthesis in Boer kids.

    PubMed

    Brassard, M-E; Chouinard, P Y; Berthiaume, R; Tremblay, G F; Gervais, R; Martineau, R; Cinq-Mars, D

    2015-11-01

    Microbial protein synthesis in the rumen would be optimized when dietary carbohydrates and proteins have synchronized rates and extent of degradation. The aim of this study was to evaluate the effect of varying ruminal degradation rate of energy and nitrogen sources on intake, nitrogen balance, microbial protein yield, and kinetics of nutrients in the rumen of growing kids. Eight Boer goats (38.2 ± 3.0 kg) were used. The treatments were arranged in a split-plot Latin square design with grain sources (barley or corn) forming the main plots (squares). Grain processing methods and levels of protein degradability formed the subplots in a 2 × 2 factorial arrangement for a total of 8 dietary treatments. The grain processing method was rolling for barley and cracking for corn. Levels of protein degradability were obtained by feeding untreated soybean meal (SBM) or heat-treated soybean meal (HSBM). Each experimental period lasted 21 d, consisting of a 10-d adaptation period, a 7-d digestibility determination period, and a 4-d rumen evacuation and sampling period. Kids fed with corn had higher purine derivatives (PD) excretion when coupled with SBM compared with HSBM and the opposite occurred with barley-fed kids ( ≤ 0.01). Unprocessed grain offered with SBM led to higher PD excretion than with HSBM whereas protein degradability had no effect when processed grain was fed ( ≤ 0.03). Results of the current experiment with high-concentrate diets showed that microbial N synthesis could be maximized in goat kids by combining slowly fermented grains (corn or unprocessed grains) with a highly degradable protein supplement (SBM). With barley, a more rapidly fermented grain, a greater microbial N synthesis was observed when supplementing a low-degradable protein (HSBM).

  4. Rumen degradable protein supply affects microbial efficiency in continuous culture and growth in steers.

    PubMed

    Brooks, M A; Harvey, R M; Johnson, N F; Kerley, M S

    2012-12-01

    We hypothesized that microbial efficiency and output from fermentation in the rumen would be optimized when peptide supply was balanced with peptide requirement of ruminal microflora. This study was conducted to measure response of varying rumen degradable peptide (RDPep) supply on ruminal fermentation characteristics and steer growth. A continuous culture experiment was conducted with diets formulated to achieve a predicted RDPep balance (RDPep supplied above RDPep required) of -0.30 to 1.45% CP with rumen degradable N (RDN) balance (RDN supplied above RDN required) above dietary ammonia-N requirement of microbes. Two additional treatments had RDPep balances of -0.30 and 0.78% CP with insufficient ammonia-N supply to meet microbial requirements. Single-flow fermenters (N = 24; n = 6) were inoculated with rumen fluid and maintained anaerobically at 39°C with a 0.06 h(-1) dilution rate. Inadequate RDN decreased OM digestion and microbial N flow, and increased rumen undegradable N (P < 0.01). Microbial efficiency decreased in RDN-deficient diets and was greatest when RDPep balance did not excessively exceed microbial requirement of RDPep predicted (P < 0.01). A growth study was conducted with 49 yearling, crossbred, Angus steers (initial BW 370 ± 34 kg). Animals were assigned to 1 of 4 treatment groups by BW and further divided into 3 pens with 4 steers per pen to achieve similar initial pen weights. Treatments consisted of 4 isonitrogenous diets balanced for RDN but varying in predicted RDPep balance (0.55%, -0.02%, -0.25%, and -0.65% CP). Animals were maintained on treatment for 70 d with individual BW taken on d 0, 1, 21, 42, 70, and 71. Final BW decreased linearly with decreasing RDPep (P = 0.05). Average daily gain and G:F displayed a quadratic effect with greater ADG and G:F at greater and lesser RDPep levels (P = 0.02). We concluded that balancing RDPep supply to predicted requirement improved fermentation efficiency and microbial output, which in turn

  5. Rumen microorganisms, methane production, and microbial protein synthesis affected by mangosteen peel powder supplement in lactating dairy cows.

    PubMed

    Polyorach, Sineenart; Wanapat, Metha; Cherdthong, Anusorn; Kang, Sungchhang

    2016-03-01

    Four crossbred dairy cows (50 % Holstein-Friesian × 50 % Thai native), 404 ± 50.0 kg of body weight (4 years old) and 90 ± 5 day in milk with daily milk production of 9 ± 2.0 kg/day, were randomly assigned according to a 4 × 4 Latin square design to study the effect of mangosteen (Garcinia mangostana) peel powder (MSP) supplementation on rumen microorganisms, methane production, and microbial protein synthesis fed concentrate containing yeast fermented cassava chip protein (YEFECAP). The treatments were different levels of MSP supplementation at 0, 100, 200, and 300 g/head/day. Rice straw was used as a roughage source fed ad libitum, and concentrate containing YEFECAP at 200 g/kg concentrate was offered corresponding to concentrate-to-milk-yield ratio at 1:2. A quantitative real-time PCR approach was used to determine the population densities of ruminal microorganisms. The results revealed that supplementation of MSP did not affect on Fibrobactor succinogenes, Ruminococcus flavefaciens, and Ruminococcus albus (P > 0.05). However, total bacteria was linearly increased (P < 0.01) while methanogens and protozoal population were linearly decreased (P < 0.01) with increasing level of MSP supplementation. Increasing level of MSP supplement could decrease rumen methane production from 27.5 to 23.7 mmol/100 ml(3). Furthermore, cows that received MSP at 300 g/head/day had the highest microbial crude protein and efficiency of rumen microbial N synthesis (416.8 g/day and 16.2 g/kg organic matter truly digested in the rumen (OMDR), respectively). In conclusion, supplementation of MSP at 300 g/head/day with YEFECAP as a protein source in the concentrate mixture revealed an enhancement of rumen fermentation and methane reduction in lactating dairy cows.

  6. Effect of diet and absence of protozoa on the rumen microbial community and on the representativeness of bacterial fractions used in the determination of microbial protein synthesis.

    PubMed

    Belanche, A; de la Fuente, G; Pinloche, E; Newbold, C J; Balcells, J

    2012-11-01

    Accurate estimates of microbial synthesis in the rumen are vital to optimize ruminant nutrition. Liquid- (LAB) and solid-associated bacterial fractions (SAB) harvested from the rumen are generally considered as microbial references when microbial yield is calculated; however, factors that determine their composition are not completely understood. The aim of this study was to evaluate the effect of diet and absence or presence of rumen protozoa on the rumen microbial community. It was hypothesized that these treatments could modify the composition and representativeness of LAB and SAB. Twenty twin lambs (Ovis aries) were used; one-half of the twins were kept protozoa-free, and each respective twin sibling was faunated. At 6 mo of age, 5 animals from each group were randomly allocated to the experimental diets consisting of either alfalfa hay as the sole diet, or 50:50 mixed with ground barley grain. After 15 d of adaptation to the diet, animals were euthanized, rumen and abomasum contents were sampled, and LAB and SAB isolated. The presence of protozoa buffered the effect of diet on the rumen bacterial population. Faunated animals fed alfalfa hay had a greater abundance of F. succinogenes, anaerobic fungi and methanogens, as well as an enhanced rumen bacterial diversity. Cellulolytic bacteria were more abundant in SAB, whereas the abomasal abundance of most of the microorganisms studied was closer to those values observed in LAB. Rumen and abomasal samples showed similar bacterial DNA concentrations, but the fungal and protozoal DNA concentration in the abomasum was only 69% and 13% of that observed in the rumen, respectively, suggesting fungal and protozoal sequestration in the rumen or possible preferential degradation of fungal and protozoal DNA in the abomasum, or both. In conclusion, absence of protozoa and type of diet extensively modified the chemical composition of LAB and SAB as a consequence of changes in the microbial composition of these fractions.

  7. Improving the quality of rice straw by urea and calcium hydroxide on rumen ecology, microbial protein synthesis in beef cattle.

    PubMed

    Polyorach, S; Wanapat, M

    2015-06-01

    Four rumen-fistulated beef cattle were randomly assigned to four treatments according to a 4 × 4 Latin square design to study the influence of urea and calcium hydroxide [Ca(OH)2 ] treatment of rice straw to improve the nutritive value of rice straw. Four dietary treatments were as follows: untreated rice straw, 50 g/kg urea-treated rice straw, 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw and 30 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw. All animals were kept in individual pens and fed with concentrate at 0.5 g/kg of BW (DM), rice straw was fed ad libitum. The experiment was conducted for four periods, and each period lasted for 21 days. During the first 14 days, DM feed intake measurements were made while during the last 7 days, all cattle were moved to metabolism crates for total faeces and urine collections. The results revealed that 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw improved the nutritive value of rice straw, in terms of dry matter intake, digestibility, ruminal volatile fatty acids, population of bacteria and fungi, nitrogen retention and microbial protein synthesis. Based on this study, it could be concluded that using urea plus calcium hydroxide was one alternative method to improve the nutritive value of rice straw, rumen ecology and fermentation and thus a reduction of treatment cost.

  8. Increasing Alfalfa Rumen Bypass Protein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Alfalfa has one of the highest crude protein contents among forage crops, but is is rapidly and extensively degraded by rumen microorganisms. To examine differential protein digestion, three distinct varieties of alfalfa, grown from single plants, were subjected to fermentation in the rumen of a ca...

  9. Effect of dietary energy source and level on nutrient digestibility, rumen microbial protein synthesis, and milk performance in lactating dairy cows.

    PubMed

    Zhou, X Q; Zhang, Y D; Zhao, M; Zhang, T; Zhu, D; Bu, D P; Wang, J Q

    2015-10-01

    This study was conducted to examine the effects of dietary energy source and level on intake, digestion, rumen microbial protein synthesis, and milk production in lactating dairy cows, using corn stover as a forage source. Eight multiparous Holstein cows, 4 of which were fitted with rumen cannulas, were evaluated in a replicated 4 × 4 Latin square design, with each period lasting 21 d. The cows were randomly assigned into 4 treatment groups: low-energy (LE) ground corn (GC), LE steam-flaked corn (SFC), high-energy (HE) GC, and HE SFC. Changes to ruminal energy degradation rates were induced by feeding the cows diets of either finely ground corn or SFC as components of diets with the same total energy level. Milk yield, milk protein content and yield, and milk lactose yield all increased in response to higher levels of dietary energy, whereas contents of milk fat and lactose were unaffected. Cows fed HE diets had a higher crude microbial protein yield and total-tract apparent digestibility than those receiving LE diets. Milk yield, milk protein yield, and microbial protein yield were also higher when SFC replaced GC as the main energy source for lactating cows fed LE diets. These results suggest that an increased dietary energy level and ruminal degradation rate are beneficial to milk protein production, which we suggest is due to increased yields of microbial proteins, when cows are fed corn stover as a dietary forage source.

  10. Rumen microbial ecology in mule deer.

    PubMed

    Pearson, H A

    1969-06-01

    Mule deer rumen microbial populations from animals in the natural habitat in Utah and from captive deer fed various rations were studied. The microorganisms were characterized on the basis of morphology and Gram reaction. Rumen samples contained 13 identifiable types of bacteria and one genus of ciliate protozoa (Entodinium). Highest rumen bacterial populations were produced on rations containing barley. No differences in proportions of ruminal bacteria in the various morphological groups could be detected when animals were fed either natural browse plants or alfalfa hay. The total numbers of bacteria were similar for animals feeding on controlled diets of browse or hay and those in the natural habitat. Numbers of some bacterial types were directly related to ciliate protozoal numbers, whereas others were inversely related. Highest rumen ciliate protozoal populations were observed on rations containing barley. No differences in protozoal populations were noted between diets containing only browse or hay. Seasonal variations were noted in ciliate protozoal numbers from deer feeding in the natural habitat. The total number of ciliate protozoa decreased in the fall and winter and remained low until spring. There were indications that salt in the deer diet favorably affected rumen ciliate protozoa. Rather than revealing direct deer management applications, this study serves to stimulate and illuminate new approaches to research in range and wildlife nutrition.

  11. Invited review: Essential oils as modifiers of rumen microbial fermentation.

    PubMed

    Calsamiglia, S; Busquet, M; Cardozo, P W; Castillejos, L; Ferret, A

    2007-06-01

    Microorganisms in the rumen degrade nutrients to produce volatile fatty acids and synthesize microbial protein as an energy and protein supply for the ruminant, respectively. However, this fermentation process has energy (losses of methane) and protein (losses of ammonia N) inefficiencies that may limit production performance and contribute to the release of pollutants to the environment. Antibiotic ionophores have been very successful in reducing these energy and protein losses in the rumen, but the use of antibiotics in animal feeds is facing reduced social acceptance, and their use has been banned in the European Union since January 2006. For this reason, scientists have become interested in evaluating other alternatives to control specific microbial populations to modulate rumen fermentation. Essential oils can interact with microbial cell membranes and inhibit the growth of some gram-positive and gram-negative bacteria. As a result of such inhibition, the addition of some plant extracts to the rumen results in an inhibition of deamination and methanogenesis, resulting in lower ammonia N, methane, and acetate, and in higher propionate and butyrate concentrations. Results have indicated that garlic oil, cinnamaldehyde (the main active component of cinnamon oil), eugenol (the main active component of the clove bud), capsaicin (the active component of hot peppers), and anise oil, among others, may increase propionate production, reduce acetate or methane production, and modify proteolysis, peptidolysis, or deamination in the rumen. However, the effects of some of these essential oils are pH and diet dependent, and their use may be beneficial only under specific conditions and production systems. For example, capsaicin appears to have small effects in high-forage diets, whereas the changes observed in high-concentrate diets (increases in dry matter intake and total VFA, and reduction in the acetateto-propionate ratio and ammonia N concentration) may be beneficial

  12. Invited review: Essential oils as modifiers of rumen microbial fermentation.

    PubMed

    Calsamiglia, S; Busquet, M; Cardozo, P W; Castillejos, L; Ferret, A

    2007-06-01

    Microorganisms in the rumen degrade nutrients to produce volatile fatty acids and synthesize microbial protein as an energy and protein supply for the ruminant, respectively. However, this fermentation process has energy (losses of methane) and protein (losses of ammonia N) inefficiencies that may limit production performance and contribute to the release of pollutants to the environment. Antibiotic ionophores have been very successful in reducing these energy and protein losses in the rumen, but the use of antibiotics in animal feeds is facing reduced social acceptance, and their use has been banned in the European Union since January 2006. For this reason, scientists have become interested in evaluating other alternatives to control specific microbial populations to modulate rumen fermentation. Essential oils can interact with microbial cell membranes and inhibit the growth of some gram-positive and gram-negative bacteria. As a result of such inhibition, the addition of some plant extracts to the rumen results in an inhibition of deamination and methanogenesis, resulting in lower ammonia N, methane, and acetate, and in higher propionate and butyrate concentrations. Results have indicated that garlic oil, cinnamaldehyde (the main active component of cinnamon oil), eugenol (the main active component of the clove bud), capsaicin (the active component of hot peppers), and anise oil, among others, may increase propionate production, reduce acetate or methane production, and modify proteolysis, peptidolysis, or deamination in the rumen. However, the effects of some of these essential oils are pH and diet dependent, and their use may be beneficial only under specific conditions and production systems. For example, capsaicin appears to have small effects in high-forage diets, whereas the changes observed in high-concentrate diets (increases in dry matter intake and total VFA, and reduction in the acetateto-propionate ratio and ammonia N concentration) may be beneficial

  13. Effect of Grape Pomace Powder, Mangosteen Peel Powder and Monensin on Nutrient Digestibility, Rumen Fermentation, Nitrogen Balance and Microbial Protein Synthesis in Dairy Steers.

    PubMed

    Foiklang, S; Wanapat, M; Norrapoke, T

    2016-10-01

    This study was designed to investigate the effect of grape pomace powder (GPP), mangosteen peel powder (MPP) and monensin on feed intake, nutrients digestibility, microorganisms, rumen fermentation characteristic, microbial protein synthesis and nitrogen balance in dairy steers. Four, rumen fistulated dairy steers with initial body weight (BW) of 220±15 kg were randomly assigned according to a 4×4 Latin square design to receive four treatments. The treatments were as follows: T1 = control, T2 = supplementation with monensin at 33 mg/kg diet, T3 = supplementation with GPP at 2% of dry matter intake, and T4 = supplementation with MPP at 30 g/kg diet. The steers were offered the concentrate diet at 0.2% BW and 3% urea treated rice straw (UTRS) was fed ad libitum. It was found that GPP supplemented group had higher UTRS intake and nutrient digestibility in terms of neutral detergent fiber and acid detergent fiber than those in control group (p<0.05). Ammonia nitrogen (NH3-N) and blood urea-nitrogen concentration were higher in monensin, GPP and MPP supplemented groups (p<0.05). Total volatile fatty acids and propionate in the GPP group were higher than those in the control group (p<0.05) while acetate concentration, and acetate to propionate ratio were decreased (p<0.01) when steers were supplemented with GPP, monensin, and MPP, respectively. Moreover, protozoal populations in GPP, MPP, and monensin supplementation were significantly lower than those in the control group (p<0.05), while cellulolytic bacterial population was significantly higher in the control group (p<0.05). Nitrogen retention, microbial crude protein and efficiency of microbial nitrogen synthesis were found significantly higher in steers that received GPP (p<0.05). Based on this study it could be concluded that the GPP has potential as an alternative feed supplement in concentrate diets which can result in improved rumen fermentation efficiency, digestibility and microbial protein synthesis in

  14. Effect of Grape Pomace Powder, Mangosteen Peel Powder and Monensin on Nutrient Digestibility, Rumen Fermentation, Nitrogen Balance and Microbial Protein Synthesis in Dairy Steers

    PubMed Central

    Foiklang, S.; Wanapat, M.; Norrapoke, T.

    2016-01-01

    This study was designed to investigate the effect of grape pomace powder (GPP), mangosteen peel powder (MPP) and monensin on feed intake, nutrients digestibility, microorganisms, rumen fermentation characteristic, microbial protein synthesis and nitrogen balance in dairy steers. Four, rumen fistulated dairy steers with initial body weight (BW) of 220±15 kg were randomly assigned according to a 4×4 Latin square design to receive four treatments. The treatments were as follows: T1 = control, T2 = supplementation with monensin at 33 mg/kg diet, T3 = supplementation with GPP at 2% of dry matter intake, and T4 = supplementation with MPP at 30 g/kg diet. The steers were offered the concentrate diet at 0.2% BW and 3% urea treated rice straw (UTRS) was fed ad libitum. It was found that GPP supplemented group had higher UTRS intake and nutrient digestibility in terms of neutral detergent fiber and acid detergent fiber than those in control group (p<0.05). Ammonia nitrogen (NH3-N) and blood urea-nitrogen concentration were higher in monensin, GPP and MPP supplemented groups (p<0.05). Total volatile fatty acids and propionate in the GPP group were higher than those in the control group (p<0.05) while acetate concentration, and acetate to propionate ratio were decreased (p<0.01) when steers were supplemented with GPP, monensin, and MPP, respectively. Moreover, protozoal populations in GPP, MPP, and monensin supplementation were significantly lower than those in the control group (p<0.05), while cellulolytic bacterial population was significantly higher in the control group (p<0.05). Nitrogen retention, microbial crude protein and efficiency of microbial nitrogen synthesis were found significantly higher in steers that received GPP (p<0.05). Based on this study it could be concluded that the GPP has potential as an alternative feed supplement in concentrate diets which can result in improved rumen fermentation efficiency, digestibility and microbial protein synthesis in

  15. Rumen microbial communities influence metabolic phenotypes in lambs

    PubMed Central

    Morgavi, Diego P.; Rathahao-Paris, Estelle; Popova, Milka; Boccard, Julien; Nielsen, Kristian F.; Boudra, Hamid

    2015-01-01

    The rumen microbiota is an essential part of ruminants shaping their nutrition and health. Despite its importance, it is not fully understood how various groups of rumen microbes affect host-microbe relationships and functions. The aim of the study was to simultaneously explore the rumen microbiota and the metabolic phenotype of lambs for identifying host-microbe associations and potential biomarkers of digestive functions. Twin lambs, separated in two groups after birth were exposed to practices (isolation and gavage with rumen fluid with protozoa or protozoa-depleted) that differentially restricted the acquisition of microbes. Rumen microbiota, fermentation parameters, digestibility and growth were monitored for up to 31 weeks of age. Microbiota assembled in isolation from other ruminants lacked protozoa and had low bacterial and archaeal diversity whereas digestibility was not affected. Exposure to adult sheep microbiota increased bacterial and archaeal diversity independently of protozoa presence. For archaea, Methanomassiliicoccales displaced Methanosphaera. Notwithstanding, protozoa induced differences in functional traits such as digestibility and significantly shaped bacterial community structure, notably Ruminococcaceae and Lachnospiraceae lower up to 6 folds, Prevotellaceae lower by ~40%, and Clostridiaceae and Veillonellaceae higher up to 10 folds compared to microbiota without protozoa. An orthogonal partial least squares-discriminant analysis of urinary metabolome matched differences in microbiota structure. Discriminant metabolites were mainly involved in amino acids and protein metabolic pathways while a negative interaction was observed between methylotrophic methanogens Methanomassiliicoccales and trimethylamine N-oxide. These results stress the influence of gut microbes on animal phenotype and show the potential of metabolomics for monitoring rumen microbial functions. PMID:26528248

  16. Rumen microbial communities influence metabolic phenotypes in lambs.

    PubMed

    Morgavi, Diego P; Rathahao-Paris, Estelle; Popova, Milka; Boccard, Julien; Nielsen, Kristian F; Boudra, Hamid

    2015-01-01

    The rumen microbiota is an essential part of ruminants shaping their nutrition and health. Despite its importance, it is not fully understood how various groups of rumen microbes affect host-microbe relationships and functions. The aim of the study was to simultaneously explore the rumen microbiota and the metabolic phenotype of lambs for identifying host-microbe associations and potential biomarkers of digestive functions. Twin lambs, separated in two groups after birth were exposed to practices (isolation and gavage with rumen fluid with protozoa or protozoa-depleted) that differentially restricted the acquisition of microbes. Rumen microbiota, fermentation parameters, digestibility and growth were monitored for up to 31 weeks of age. Microbiota assembled in isolation from other ruminants lacked protozoa and had low bacterial and archaeal diversity whereas digestibility was not affected. Exposure to adult sheep microbiota increased bacterial and archaeal diversity independently of protozoa presence. For archaea, Methanomassiliicoccales displaced Methanosphaera. Notwithstanding, protozoa induced differences in functional traits such as digestibility and significantly shaped bacterial community structure, notably Ruminococcaceae and Lachnospiraceae lower up to 6 folds, Prevotellaceae lower by ~40%, and Clostridiaceae and Veillonellaceae higher up to 10 folds compared to microbiota without protozoa. An orthogonal partial least squares-discriminant analysis of urinary metabolome matched differences in microbiota structure. Discriminant metabolites were mainly involved in amino acids and protein metabolic pathways while a negative interaction was observed between methylotrophic methanogens Methanomassiliicoccales and trimethylamine N-oxide. These results stress the influence of gut microbes on animal phenotype and show the potential of metabolomics for monitoring rumen microbial functions. PMID:26528248

  17. Improving rumen ecology and microbial population by dried rumen digesta in beef cattle.

    PubMed

    Cherdthong, Anusorn; Wanapat, Metha; Saenkamsorn, Anuthida; Supapong, Chanadol; Anantasook, Nirawan; Gunun, Pongsatorn

    2015-06-01

    Four Thai native beef cattle with initial body weight (BW) of 91.8 ± 4.75 kg were randomly assigned according to a 4 × 4 Latin square design to receive four concentrates replacement levels of soybean meal (SBM) by dried rumen digesta (DRD) at 0, 33, 67, and 100 % on dry matter (DM) basis. All cattle were fed rice straw ad libitum while additional concentrate was fed at 0.5 % BW daily. The experiment was conducted for four periods of 21 days. Rumen fluid was analyzed for predominant cellulolytic bacterial population by using real-time PCR technique. Increasing levels of DRD did not alter total feed intake, ruminal pH and temperature, and plasma urea nitrogen (P > 0.05). Protozoa and fungal population were not differed by DRD supplementation while population of bacteria at 4 h post feeding was increased when SBM was replaced with DRD at 66 and 100 % DM. Population of total bacteria and R. flavefaciens at 4 h post feeding were significantly highest with inclusion of 100 % of DRD in the ration. The experimental diets has no effect on excretion and absorption of purine derivatives (P > 0.05), while microbial crude protein and efficiency of microbial N synthesis were significantly increased with DRD inclusion in the diet and highest with 100 % DRD replacement (P > 0.05). Replacement of SBM by DRD at 100 % DM improved the rumen ecology and microbial population in beef cattle fed on rice straw.

  18. The importance of the form of nitrogen on microbial protein synthesis in the rumen of cattle receiving grass silage and continuous intrarumen infusions of sucrose.

    PubMed

    Rooke, J A; Armstrong, D G

    1989-01-01

    1. In a 4 x 4 Latin square design experiment, four cattle were given grass silage in two meals per d to satisfy maintenance energy requirements. In addition, sucrose (170 g/kg silage dry matter (DM] was infused intraruminally at a constant rate with no nitrogen supplementation; with the infusion intraruminally of either casein (23 g/kg silage DM) or urea (8 g/kg silage DM); or with soya-bean meal (64 g/kg silage DM) fed in two equal portions. 2. Samples of duodenal digesta representative of a 24 h period were obtained using chromium-EDTA and ytterbium acetate for flow estimation and 35S as a marker of microbial N entering the small intestine. Samples of rumen fluid were also taken for estimation of rumen pH and concentrations of ammonia-N and volatile fatty acids. Estimates of apparent organic matter (OM) and N digestibility and of the rates of silage DM and N disappearance from porous synthetic-fibre bags incubated in the rumen were also made. 3. The N supplements had no significant effects on rumen pH, concentrations of volatile fatty acids, their molar proportions or the disappearance of DM or N from porous synthetic-fibre bags. N supplementation increased rumen ammonia-N concentrations (urea, P less than 0.05; casein, soya-bean meal, not significant). 4. N supplementation had no significant effects on the digestion of OM, acid-detergent fibre or soluble carbohydrate. 5. Infusion of casein increased the quantities of total non-ammonia-N (not significant) and microbial N (P less than 0.05) entering the small intestine daily and the efficiency of rumen microbial N synthesis (not significant).(ABSTRACT TRUNCATED AT 250 WORDS)

  19. Rumen microbial (meta)genomics and its application to ruminant production.

    PubMed

    Morgavi, D P; Kelly, W J; Janssen, P H; Attwood, G T

    2013-03-01

    Meat and milk produced by ruminants are important agricultural products and are major sources of protein for humans. Ruminant production is of considerable economic value and underpins food security in many regions of the world. However, the sector faces major challenges because of diminishing natural resources and ensuing increases in production costs, and also because of the increased awareness of the environmental impact of farming ruminants. The digestion of feed and the production of enteric methane are key functions that could be manipulated by having a thorough understanding of the rumen microbiome. Advances in DNA sequencing technologies and bioinformatics are transforming our understanding of complex microbial ecosystems, including the gastrointestinal tract of mammals. The application of these techniques to the rumen ecosystem has allowed the study of the microbial diversity under different dietary and production conditions. Furthermore, the sequencing of genomes from several cultured rumen bacterial and archaeal species is providing detailed information about their physiology. More recently, metagenomics, mainly aimed at understanding the enzymatic machinery involved in the degradation of plant structural polysaccharides, is starting to produce new insights by allowing access to the total community and sidestepping the limitations imposed by cultivation. These advances highlight the promise of these approaches for characterising the rumen microbial community structure and linking this with the functions of the rumen microbiota. Initial results using high-throughput culture-independent technologies have also shown that the rumen microbiome is far more complex and diverse than the human caecum. Therefore, cataloguing its genes will require a considerable sequencing and bioinformatic effort. Nevertheless, the construction of a rumen microbial gene catalogue through metagenomics and genomic sequencing of key populations is an attainable goal. A rumen

  20. Rumen microbial (meta)genomics and its application to ruminant production.

    PubMed

    Morgavi, D P; Kelly, W J; Janssen, P H; Attwood, G T

    2013-03-01

    Meat and milk produced by ruminants are important agricultural products and are major sources of protein for humans. Ruminant production is of considerable economic value and underpins food security in many regions of the world. However, the sector faces major challenges because of diminishing natural resources and ensuing increases in production costs, and also because of the increased awareness of the environmental impact of farming ruminants. The digestion of feed and the production of enteric methane are key functions that could be manipulated by having a thorough understanding of the rumen microbiome. Advances in DNA sequencing technologies and bioinformatics are transforming our understanding of complex microbial ecosystems, including the gastrointestinal tract of mammals. The application of these techniques to the rumen ecosystem has allowed the study of the microbial diversity under different dietary and production conditions. Furthermore, the sequencing of genomes from several cultured rumen bacterial and archaeal species is providing detailed information about their physiology. More recently, metagenomics, mainly aimed at understanding the enzymatic machinery involved in the degradation of plant structural polysaccharides, is starting to produce new insights by allowing access to the total community and sidestepping the limitations imposed by cultivation. These advances highlight the promise of these approaches for characterising the rumen microbial community structure and linking this with the functions of the rumen microbiota. Initial results using high-throughput culture-independent technologies have also shown that the rumen microbiome is far more complex and diverse than the human caecum. Therefore, cataloguing its genes will require a considerable sequencing and bioinformatic effort. Nevertheless, the construction of a rumen microbial gene catalogue through metagenomics and genomic sequencing of key populations is an attainable goal. A rumen

  1. Microbial numbers, rumen fermentation, and nitrogen utilization of steers fed wet or dried brewers' grains.

    PubMed

    Rogers, J A; Conrad, H R; Dehority, B A; Grubb, J A

    1986-03-01

    Holstein steers were fed corn silage supplemented with either wet or dried brewers' grains to determine effects of heat drying commercial brewers' grains. Four rumen-fistulated steers were fed a 12.5% crude protein diet in a single reversal design experiment. Brewers' grains supplied 45% of the protein of the diet. Bacterial numbers, concentration of ciliated protozoa, and ammonia concentration in the rumen were higher, and rumen pH was lower, for steers fed wet brewers' grains. Concentrations of rumen volatile fatty acids were similar for both diets. Ruminal digestibility of dry matter decreased when wet versus dried brewers' grains were fed (56.9 versus 39.3%). The rate of dry matter passage from the rumen was faster with wet brewers' grains. In Experiment 2, 12 steers were in a 2 X 2 factorial design. Diets contained wet or dried brewers' grains supplemented at 22 or 40% of the diet dry matter (12.5 and 14.5% crude protein). Nitrogen retention was increased in steers fed the higher crude protein diet. Apparent digestible nitrogen, acid detergent fiber nitrogen, and nitrogen retention were higher with wet versus dried brewers' grains. Plasma essential and nonessential amino acids were also higher in steers fed wet brewers' grains. Alteration in microbial numbers, fermentation measurements, and nitrogen utilization were associated with more soluble nitrogen with wet (13.4%) versus dried (3.3%) brewers' grains.

  2. Effects of quebracho tannin extract (Schinopsis balansae Engl.) and activated charcoal on nitrogen balance, rumen microbial protein synthesis and faecal composition of growing Boer goats.

    PubMed

    Al-Kindi, Amal; Dickhoefer, Uta; Schlecht, Eva; Sundrum, Albert; Schiborra, Anne

    2016-08-01

    Under irrigated arid conditions, organic fertiliser rich in slowly decomposable nitrogen (N) and carbon (C) is needed for soil fertility maintenance. Feeding ruminants with condensed tannins will lower ruminal protein degradation, reduce urinary N excretion and might increase the faecal fraction of slowly decomposable N. Supplementation with activated charcoal (AC) might enrich manure with slowly degrading C. Therefore, we investigated the effects of feeding quebracho tannin extract (QTE) and AC on the N balance of goats, the efficiency of microbial protein synthesis in the rumen (EMPS) and the composition of faeces. The feeding trial comprised three periods; in each period, 12 male Boer goats (28 ± 3.9 kg live weight) were assigned to six treatments: a Control diet (per kg diet 500 g grass hay and 500 g concentrate) and to further five treatments the Control diet was supplemented with QTE (20 g and 40 g/kg; diets QTE2 and QTE4, respectively), with AC (15 g and 30 g/kg, diets AC1.5 and AC3.0, respectively) and a mixture of QTE (20 g/kg) plus AC (15 g/kg) (diet QTEAC). In addition to the N balance, EMPS was calculated from daily excretions of purine derivatives, and the composition of faecal N was determined. There was no effect of QTE and AC supplementation on the intake of organic matter (OM), N and fibre, but apparent total tract digestibility of OM was reduced (p = 0.035). Feeding QTE induced a shift in N excretion from urine to faeces (p ≤ 0.001) without altering N retention. Total N excretion tended to decrease with QTE treatments (p = 0.053), but EMPS was not different between treatments. Faecal C excretion was higher in QTE and AC treatments (p = 0.001) compared with the Control, while the composition of faecal N differed only in concentration of undigested dietary N (p = 0.001). The results demonstrate that QTE can be included into diets of goats up to 40 g/kg, without affecting N utilisation, but simultaneously increasing the

  3. Effects of quebracho tannin extract (Schinopsis balansae Engl.) and activated charcoal on nitrogen balance, rumen microbial protein synthesis and faecal composition of growing Boer goats.

    PubMed

    Al-Kindi, Amal; Dickhoefer, Uta; Schlecht, Eva; Sundrum, Albert; Schiborra, Anne

    2016-08-01

    Under irrigated arid conditions, organic fertiliser rich in slowly decomposable nitrogen (N) and carbon (C) is needed for soil fertility maintenance. Feeding ruminants with condensed tannins will lower ruminal protein degradation, reduce urinary N excretion and might increase the faecal fraction of slowly decomposable N. Supplementation with activated charcoal (AC) might enrich manure with slowly degrading C. Therefore, we investigated the effects of feeding quebracho tannin extract (QTE) and AC on the N balance of goats, the efficiency of microbial protein synthesis in the rumen (EMPS) and the composition of faeces. The feeding trial comprised three periods; in each period, 12 male Boer goats (28 ± 3.9 kg live weight) were assigned to six treatments: a Control diet (per kg diet 500 g grass hay and 500 g concentrate) and to further five treatments the Control diet was supplemented with QTE (20 g and 40 g/kg; diets QTE2 and QTE4, respectively), with AC (15 g and 30 g/kg, diets AC1.5 and AC3.0, respectively) and a mixture of QTE (20 g/kg) plus AC (15 g/kg) (diet QTEAC). In addition to the N balance, EMPS was calculated from daily excretions of purine derivatives, and the composition of faecal N was determined. There was no effect of QTE and AC supplementation on the intake of organic matter (OM), N and fibre, but apparent total tract digestibility of OM was reduced (p = 0.035). Feeding QTE induced a shift in N excretion from urine to faeces (p ≤ 0.001) without altering N retention. Total N excretion tended to decrease with QTE treatments (p = 0.053), but EMPS was not different between treatments. Faecal C excretion was higher in QTE and AC treatments (p = 0.001) compared with the Control, while the composition of faecal N differed only in concentration of undigested dietary N (p = 0.001). The results demonstrate that QTE can be included into diets of goats up to 40 g/kg, without affecting N utilisation, but simultaneously increasing the

  4. Microbial and chemical composition of liquid-associated bacteria in goats' rumen and fermenters.

    PubMed

    Abecia, L; Soto, E C; Ramos-Morales, E; Molina-Alcaide, E

    2014-10-01

    This study was undertaken to investigate the relationship between chemical composition and microbial profile of rumen liquid-associated bacteria (LAB) in vivo (Murciano-Granadina goats) and in a rumen simulation system (single-flow continuous-culture fermenters). To achieve this aim, analyses of purine bases along with some molecular techniques (quantitative PCR to assess abundance and DGGE to identify biodiversity and bacterial profile) were carried out. A control diet (AHC) based on alfalfa hay (AH) and concentrate (C) in a 1:1 ratio and two experimental diets (AHCBI and AHCBII), in which concentrate was partially replaced with multinutrient blocks, were used. Diets AHCBI and AHCBII included multinutrient blocks differing in the relative amount of two-stage olive cake and the source of protein (sunflower meal vs. fava beans). We aimed to investigate the effect of these blocks on rumen microbiota to evaluate their potential as safe substitutes of cereal-based concentrates. Similar patterns of response to diet were found for chemical composition, microbial abundances and diversity in LAB isolated from goat's rumen and fermenters. Whereas bacterial density (log10 gene copies/g FM: 11.6 and 9.4 for bacteria and methanogens, respectively, in rumen) and diversity indexes (Shannon index: 3.6) were not affected by diet, DGGE analyses showed that bacterial community profile was affected. The cluster analysis suggested differences in bacterial profile between LAB pellets isolated from the rumen of goat and fermenters. A relationship between chemical composition and bacterial community composition in LAB pellets seems to exist. Changes in the former were reflected in the bacterial community profile. Further research is needed to clarify the relationship between chemical and microbial composition of ruminal bacterial pellets with diets of different quality.

  5. Microbial and chemical composition of liquid-associated bacteria in goats' rumen and fermenters.

    PubMed

    Abecia, L; Soto, E C; Ramos-Morales, E; Molina-Alcaide, E

    2014-10-01

    This study was undertaken to investigate the relationship between chemical composition and microbial profile of rumen liquid-associated bacteria (LAB) in vivo (Murciano-Granadina goats) and in a rumen simulation system (single-flow continuous-culture fermenters). To achieve this aim, analyses of purine bases along with some molecular techniques (quantitative PCR to assess abundance and DGGE to identify biodiversity and bacterial profile) were carried out. A control diet (AHC) based on alfalfa hay (AH) and concentrate (C) in a 1:1 ratio and two experimental diets (AHCBI and AHCBII), in which concentrate was partially replaced with multinutrient blocks, were used. Diets AHCBI and AHCBII included multinutrient blocks differing in the relative amount of two-stage olive cake and the source of protein (sunflower meal vs. fava beans). We aimed to investigate the effect of these blocks on rumen microbiota to evaluate their potential as safe substitutes of cereal-based concentrates. Similar patterns of response to diet were found for chemical composition, microbial abundances and diversity in LAB isolated from goat's rumen and fermenters. Whereas bacterial density (log10 gene copies/g FM: 11.6 and 9.4 for bacteria and methanogens, respectively, in rumen) and diversity indexes (Shannon index: 3.6) were not affected by diet, DGGE analyses showed that bacterial community profile was affected. The cluster analysis suggested differences in bacterial profile between LAB pellets isolated from the rumen of goat and fermenters. A relationship between chemical composition and bacterial community composition in LAB pellets seems to exist. Changes in the former were reflected in the bacterial community profile. Further research is needed to clarify the relationship between chemical and microbial composition of ruminal bacterial pellets with diets of different quality. PMID:24460876

  6. Shifts in the rumen microbiota due to the type of carbohydrate and level of protein ingested by dairy cattle are associated with changes in rumen fermentation.

    PubMed

    Belanche, Alejandro; Doreau, Michel; Edwards, Joan E; Moorby, Jon M; Pinloche, Eric; Newbold, Charles J

    2012-09-01

    Balancing energy and nitrogen in the rumen is a key to both profitability and environmental sustainability. Four dairy cows were used in a Latin square experimental design to investigate the effect of severe nitrogen underfeeding (110 vs. 80% of requirements) and the type of carbohydrate consumed [neutral detergent fiber rich (FIB) vs. starch rich (STA)] on the rumen ecosystem. These dietary treatments modified both rumen fermentation and microbial populations. Compared with STA diets, consumption of FIB diets increased bacterial and fungal diversity in the rumen and also increased the concentrations of cellulolytic microorganisms, including protozoa (+38%), anaerobic fungi (+59%), and methanogens (+27%). This microbial adaptation to fiber utilization led to similar digestibility values for the 2 carbohydrate sources and was accompanied by a shift in the rumen fermentation patterns; when the FIB diets were consumed, the cows had greater ruminal pH, ammonia concentrations, and molar proportions of acetate and propionate compared with when they consumed the STA diets. Certain rumen microorganisms were sensitive to a shortage of nitrogen; rumen concentrations of ammonia were 49% lower when the low-protein (LP) diets were consumed as were total bacteria (-13%), anaerobic fungi (-28%), methanogens (-27%), protozoa (-19%), cellulolytic bacteria, and microbial diversity compared with when the high-protein (HP) diets were consumed. As a result, the digestibility of the LP diets was less than that of the HP diets. These findings demonstrated that the rumen microbial ecosystem is directly linked to the rumen fermentation pattern and, to some extent, to the efficiency of diet utilization by dairy cattle.

  7. Evaluation of DNA extraction methods of rumen microbial populations.

    PubMed

    Villegas-Rivera, Gabriela; Vargas-Cabrera, Yevani; González-Silva, Napoleón; Aguilera-García, Florentino; Gutiérrez-Vázquez, Ernestina; Bravo-Patiño, Alejandro; Cajero-Juárez, Marcos; Baizabal-Aguirre, Víctor Manuel; Valdez-Alarcón, Juan José

    2013-02-01

    The dynamism of microbial populations in the rumen has been studied with molecular methods that analyze single nucleotide polymorphisms of ribosomal RNA gene fragments (rDNA). Therefore DNA of good quality is needed for this kind of analysis. In this work we report the evaluation of four DNA extraction protocols (mechanical lysis or chemical lysis with CTAB, ethylxanthogenate or DNAzol(®)) from ruminal fluid. The suitability of two of these protocols (mechanical lysis and DNAzol(®)) was tested on single-strand conformation polymorphism (SSCP) of rDNA of rumen microbial populations. DNAzol(®) was a simple method that rendered good integrity, yield and purity. With this method, subtle changes in protozoan populations were detected in young bulls fed with slightly different formulations of a supplement of multinutritional blocks of molasses and urea. Sequences related to Eudiplodinium maggi and a non-cultured Entodiniomorphid similar to Entodinium caudatum, were related to major fluctuating populations in an SSCP assay.

  8. Board-invited review: Rumen microbiology: Leading the way in microbial ecology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Robert Hungate, considered the father of rumen microbiology, was the first to initiate a systematic exploration of the microbial ecosystem of the rumen, but he was not alone. The techniques he developed to isolate and identify cellulose-digesting bacteria from the rumen have had a major impact not ...

  9. Immunization against Rumen Methanogenesis by Vaccination with a New Recombinant Protein.

    PubMed

    Zhang, Litai; Huang, Xiaofeng; Xue, Bai; Peng, Quanhui; Wang, Zhisheng; Yan, Tianhai; Wang, Lizhi

    2015-01-01

    Vaccination through recombinant proteins against rumen methanogenesis provides a mitigation approach to reduce enteric methane (CH4) emissions in ruminants. The objective of present study was to evaluate the in vivo efficacy of a new vaccine candidate protein (EhaF) on methanogenesis and microbial population in the rumen of goats. We amplified the gene mru 1407 encoding protein EhaF using fresh rumen fluid samples of mature goats and successfully expressed recombinant protein (EhaF) in Escherichia coli Rosetta. This product was evaluated using 12 mature goats with half for control and other half injected with 400ug/goat the purified recombinant protein in day 1 and two subsequent booster immunizations in day 35 and 49. All measurements were undertaken from 63 to 68 days after the initial vaccination, with CH4 emissions determined using respiration calorimeter chambers. The results showed that the vaccination caused intensive immune responses in serum and saliva, although it had no significant effect on total enteric CH4 emissions and methanogen population in the rumen, when compared with the control goats. However, the vaccination altered the composition of rumen bacteria, especially the abundance of main phylum Firmicutes and genus Prevotella. The results indicate that protein EhaF might not be an effective vaccine to reduce enteric CH4 emissions but our vaccine have potential to influence the rumen ecosystem of goats. PMID:26445479

  10. Immunization against Rumen Methanogenesis by Vaccination with a New Recombinant Protein

    PubMed Central

    Zhang, Litai; Huang, Xiaofeng; Xue, Bai; Peng, Quanhui; Wang, Zhisheng; Yan, Tianhai; Wang, Lizhi

    2015-01-01

    Vaccination through recombinant proteins against rumen methanogenesis provides a mitigation approach to reduce enteric methane (CH4) emissions in ruminants. The objective of present study was to evaluate the in vivo efficacy of a new vaccine candidate protein (EhaF) on methanogenesis and microbial population in the rumen of goats. We amplified the gene mru 1407 encoding protein EhaF using fresh rumen fluid samples of mature goats and successfully expressed recombinant protein (EhaF) in Escherichia coli Rosetta. This product was evaluated using 12 mature goats with half for control and other half injected with 400ug/goat the purified recombinant protein in day 1 and two subsequent booster immunizations in day 35 and 49. All measurements were undertaken from 63 to 68 days after the initial vaccination, with CH4 emissions determined using respiration calorimeter chambers. The results showed that the vaccination caused intensive immune responses in serum and saliva, although it had no significant effect on total enteric CH4 emissions and methanogen population in the rumen, when compared with the control goats. However, the vaccination altered the composition of rumen bacteria, especially the abundance of main phylum Firmicutes and genus Prevotella. The results indicate that protein EhaF might not be an effective vaccine to reduce enteric CH4 emissions but our vaccine have potential to influence the rumen ecosystem of goats. PMID:26445479

  11. Immunization against Rumen Methanogenesis by Vaccination with a New Recombinant Protein.

    PubMed

    Zhang, Litai; Huang, Xiaofeng; Xue, Bai; Peng, Quanhui; Wang, Zhisheng; Yan, Tianhai; Wang, Lizhi

    2015-01-01

    Vaccination through recombinant proteins against rumen methanogenesis provides a mitigation approach to reduce enteric methane (CH4) emissions in ruminants. The objective of present study was to evaluate the in vivo efficacy of a new vaccine candidate protein (EhaF) on methanogenesis and microbial population in the rumen of goats. We amplified the gene mru 1407 encoding protein EhaF using fresh rumen fluid samples of mature goats and successfully expressed recombinant protein (EhaF) in Escherichia coli Rosetta. This product was evaluated using 12 mature goats with half for control and other half injected with 400ug/goat the purified recombinant protein in day 1 and two subsequent booster immunizations in day 35 and 49. All measurements were undertaken from 63 to 68 days after the initial vaccination, with CH4 emissions determined using respiration calorimeter chambers. The results showed that the vaccination caused intensive immune responses in serum and saliva, although it had no significant effect on total enteric CH4 emissions and methanogen population in the rumen, when compared with the control goats. However, the vaccination altered the composition of rumen bacteria, especially the abundance of main phylum Firmicutes and genus Prevotella. The results indicate that protein EhaF might not be an effective vaccine to reduce enteric CH4 emissions but our vaccine have potential to influence the rumen ecosystem of goats.

  12. Plant extracts affect in vitro rumen microbial fermentation.

    PubMed

    Busquet, M; Calsamiglia, S; Ferret, A; Kamel, C

    2006-02-01

    Different doses of 12 plant extracts and 6 secondary plant metabolites were incubated for 24 h in diluted ruminal fluid with a 50:50 forage:concentrate diet. Treatments were: control (no additive), plant extracts (anise oil, cade oil, capsicum oil, cinnamon oil, clove bud oil, dill oil, fenugreek, garlic oil, ginger oil, oregano oil, tea tree oil, and yucca), and secondary plant metabolites (anethol, benzyl salicylate, carvacrol, carvone, cinnamaldehyde, and eugenol). Each treatment was supplied at 3, 30, 300, and 3,000 mg/L of culture fluid. At 3,000 mg/L, most treatments decreased total volatile fatty acid concentration, but cade oil, capsicum oil, dill oil, fenugreek, ginger oil, and yucca had no effect. Different doses of anethol, anise oil, carvone, and tea tree oil decreased the proportion of acetate and propionate, which suggests that these compounds may not be nutritionally beneficial to dairy cattle. Garlic oil (300 and 3,000 mg/L) and benzyl salicylate (300 and 3,000 mg/L) reduced acetate and increased propionate and butyrate proportions, suggesting that methane production was inhibited. At 3,000 mg/L, capsicum oil, carvacrol, carvone, cinnamaldehyde, cinnamon oil, clove bud oil, eugenol, fenugreek, and oregano oil resulted in a 30 to 50% reduction in ammonia N concentration. Careful selection and combination of these extracts may allow the manipulation of rumen microbial fermentation.

  13. Board-invited review: Rumen microbiology: leading the way in microbial ecology.

    PubMed

    Krause, D O; Nagaraja, T G; Wright, A D G; Callaway, T R

    2013-01-01

    Robert Hungate, considered the father of rumen microbiology, was the first to initiate a systematic exploration of the microbial ecosystem of the rumen, but he was not alone. The techniques he developed to isolate and identify cellulose-digesting bacteria from the rumen have had a major impact not only in delineating the complex ecosystem of the rumen but also in clinical microbiology and in the exploration of a number of other anaerobic ecosystems, including the human hindgut. Rumen microbiology has pioneered our understanding of much of microbial ecology and has broadened our knowledge of ecology in general, as well as improved the ability to feed ruminants more efficiently. The discovery of anaerobic fungi as a component of the ruminal flora disproved the central dogma in microbiology that all fungi are aerobic organisms. Further novel interactions between bacterial species such as nutrient cross feeding and interspecies H2 transfer were first described in ruminal microorganisms. The complexity and diversity present in the rumen make it an ideal testing ground for microbial theories (e.g., the effects of nutrient limitation and excess) and techniques (such as 16S rRNA), which have rewarded the investigators that have used this easily accessed ecosystem to understand larger truths. Our understanding of characteristics of the ruminal microbial population has opened new avenues of microbial ecology, such as the existence of hyperammonia-producing bacteria and how they can be used to improve N efficiency in ruminants. In this review, we examine some of the contributions to science that were first made in the rumen, which have not been recognized in a broader sense.

  14. Methane Inhibition Alters the Microbial Community, Hydrogen Flow, and Fermentation Response in the Rumen of Cattle.

    PubMed

    Martinez-Fernandez, Gonzalo; Denman, Stuart E; Yang, Chunlei; Cheung, Jane; Mitsumori, Makoto; McSweeney, Christopher S

    2016-01-01

    Management of metabolic hydrogen ([H]) in the rumen has been identified as an important consideration when reducing ruminant CH4 emissions. However, little is known about hydrogen flux and microbial rumen population responses to CH4 inhibition when animals are fed with slowly degradable diets. The effects of the anti-methanogenic compound, chloroform, on rumen fermentation, microbial ecology, and H2/CH4 production were investigated in vivo. Eight rumen fistulated Brahman steers were fed a roughage hay diet (Rhode grass hay) or roughage hay:concentrate diet (60:40) with increasing levels (low, mid, and high) of chloroform in a cyclodextrin matrix. The increasing levels of chloroform resulted in an increase in H2 expelled as CH4 production decreased with no effect on dry matter intakes. The amount of expelled H2 per mole of decreased methane, was lower for the hay diet suggesting a more efficient redirection of hydrogen into other microbial products compared with hay:concentrate diet. A shift in rumen fermentation toward propionate and branched-chain fatty acids was observed for both diets. Animals fed with the hay:concentrate diet had both higher formate concentration and H2 expelled than those fed only roughage hay. Metabolomic analyses revealed an increase in the concentration of amino acids, organic, and nucleic acids in the fluid phase for both diets when methanogenesis was inhibited. These changes in the rumen metabolism were accompanied by a shift in the microbiota with an increase in Bacteroidetes:Firmicutes ratio and a decrease in Archaea and Synergistetes for both diets. Within the Bacteroidetes family, some OTUs assigned to Prevotella were promoted under chloroform treatment. These bacteria may be partly responsible for the increase in amino acids and propionate in the rumen. No significant changes were observed for abundance of fibrolytic bacteria, protozoa, and fungi, which suggests that fiber degradation was not impaired. The observed 30% decrease in

  15. Methane Inhibition Alters the Microbial Community, Hydrogen Flow, and Fermentation Response in the Rumen of Cattle.

    PubMed

    Martinez-Fernandez, Gonzalo; Denman, Stuart E; Yang, Chunlei; Cheung, Jane; Mitsumori, Makoto; McSweeney, Christopher S

    2016-01-01

    Management of metabolic hydrogen ([H]) in the rumen has been identified as an important consideration when reducing ruminant CH4 emissions. However, little is known about hydrogen flux and microbial rumen population responses to CH4 inhibition when animals are fed with slowly degradable diets. The effects of the anti-methanogenic compound, chloroform, on rumen fermentation, microbial ecology, and H2/CH4 production were investigated in vivo. Eight rumen fistulated Brahman steers were fed a roughage hay diet (Rhode grass hay) or roughage hay:concentrate diet (60:40) with increasing levels (low, mid, and high) of chloroform in a cyclodextrin matrix. The increasing levels of chloroform resulted in an increase in H2 expelled as CH4 production decreased with no effect on dry matter intakes. The amount of expelled H2 per mole of decreased methane, was lower for the hay diet suggesting a more efficient redirection of hydrogen into other microbial products compared with hay:concentrate diet. A shift in rumen fermentation toward propionate and branched-chain fatty acids was observed for both diets. Animals fed with the hay:concentrate diet had both higher formate concentration and H2 expelled than those fed only roughage hay. Metabolomic analyses revealed an increase in the concentration of amino acids, organic, and nucleic acids in the fluid phase for both diets when methanogenesis was inhibited. These changes in the rumen metabolism were accompanied by a shift in the microbiota with an increase in Bacteroidetes:Firmicutes ratio and a decrease in Archaea and Synergistetes for both diets. Within the Bacteroidetes family, some OTUs assigned to Prevotella were promoted under chloroform treatment. These bacteria may be partly responsible for the increase in amino acids and propionate in the rumen. No significant changes were observed for abundance of fibrolytic bacteria, protozoa, and fungi, which suggests that fiber degradation was not impaired. The observed 30% decrease in

  16. Methane Inhibition Alters the Microbial Community, Hydrogen Flow, and Fermentation Response in the Rumen of Cattle

    PubMed Central

    Martinez-Fernandez, Gonzalo; Denman, Stuart E.; Yang, Chunlei; Cheung, Jane; Mitsumori, Makoto; McSweeney, Christopher S.

    2016-01-01

    Management of metabolic hydrogen ([H]) in the rumen has been identified as an important consideration when reducing ruminant CH4 emissions. However, little is known about hydrogen flux and microbial rumen population responses to CH4 inhibition when animals are fed with slowly degradable diets. The effects of the anti-methanogenic compound, chloroform, on rumen fermentation, microbial ecology, and H2/CH4 production were investigated in vivo. Eight rumen fistulated Brahman steers were fed a roughage hay diet (Rhode grass hay) or roughage hay:concentrate diet (60:40) with increasing levels (low, mid, and high) of chloroform in a cyclodextrin matrix. The increasing levels of chloroform resulted in an increase in H2 expelled as CH4 production decreased with no effect on dry matter intakes. The amount of expelled H2 per mole of decreased methane, was lower for the hay diet suggesting a more efficient redirection of hydrogen into other microbial products compared with hay:concentrate diet. A shift in rumen fermentation toward propionate and branched-chain fatty acids was observed for both diets. Animals fed with the hay:concentrate diet had both higher formate concentration and H2 expelled than those fed only roughage hay. Metabolomic analyses revealed an increase in the concentration of amino acids, organic, and nucleic acids in the fluid phase for both diets when methanogenesis was inhibited. These changes in the rumen metabolism were accompanied by a shift in the microbiota with an increase in Bacteroidetes:Firmicutes ratio and a decrease in Archaea and Synergistetes for both diets. Within the Bacteroidetes family, some OTUs assigned to Prevotella were promoted under chloroform treatment. These bacteria may be partly responsible for the increase in amino acids and propionate in the rumen. No significant changes were observed for abundance of fibrolytic bacteria, protozoa, and fungi, which suggests that fiber degradation was not impaired. The observed 30% decrease in

  17. Low protein silage associated with rumen impaction in suckler cows.

    PubMed

    2016-04-23

    Rumen impaction associated with low protein diets in a suckler cowCampylobacteriosis in suckler cowsPlant toxicity in ewesListerial encephalitis in ewes ITALIC! Chorioptes bovis-associated infertility in ramsThese are among matters discussed in the disease surveillance report for January 2016 from SAC Consulting: Veterinary Services (SAC C VS).

  18. Effect of Various Essential Oils Isolated from Douglas Fir Needles upon Sheep and Deer Rumen Microbial Activity

    PubMed Central

    Oh, Hi Kon; Sakai, T.; Jones, M. B.; Longhurst, W. M.

    1967-01-01

    The effects of essential oils isolated from Douglas fir needles on sheep and deer rumen microbial activity were tested by use of an anaerobic manometric technique. Rumen microorganisms were obtained from a sheep which had been fed mainly on alfalfa hay and dried range grass. One deer used in this study had access to Douglas fir trees the year around, whereas the other deer had no access to Douglas fir. All of the monoterpene hydrocarbons isolated from Douglas fir needles—α-pinene, β-pinene, limonene, myrcene, camphene, Δ3-carene, and terpinolene—promoted only slightly or had no effect on deer rumen microbial activity, whereas all of them promoted activity in sheep rumen microbes, except Δ3-carene and terpinolene, which inhibited activity. Of the oxygenated monoterpenes, all monoterpene alcohols—α-terpineol, terpinen-4-ol, linalool, citronellol, and fenchyl alcohol—strongly inhibited the rumen microbial activity of both sheep and deer. Monoterpene esters (bornyl acetate) produced mild inhibition for both sheep and deer microbes, and citronellyl acetate inhibited rumen microbial activity in sheep, whereas it promoted activity in both deer. Monoterpene aldehyde (citronellal) inhibited the activity of rumen microbes from both sheep and deer having no access to Douglas fir from the Hopland Field Station, whereas they produced no effect upon the deer having access to Douglas fir from the Masonite forest. Rumen microbial activity for sheep and deer was promoted slightly with aliphatic ester (ethyl-n-caproate). There was a marked difference between sheep and deer rumen microbes as affected by addition of the various essential oils. The monoterpene hydrocarbons promoted activity more on sheep rumen microbes than on deer, and the monoterpene alcohols inhibited sheep rumen microbial activity more than that of deer. Furthermore, the deer rumen microbes from Hopland Field Station were affected more than the deer from Masonite forest. Images Fig. 1 PMID:6049303

  19. Rumen transfaunation.

    PubMed

    DePeters, E J; George, L W

    2014-12-01

    The aim of this invited mini-review is to summarize the rumen transfaunation literature. Rumen transfaunation using the cud from a healthy donor animal to treat a sick recipient animal was practiced long before our understanding of rumen microorganisms. Around the mid-1900 s, scientists began to explore the benefits of rumen transfaunation and the associated microbial populations. Rumen transfaunation has been used clinically to treat indigestion and to enhance the return of normal rumen function following surgical correction of a left-displaced abomasum. Rumen transfaunation was also used to introduce unique rumen microorganisms into animals that were exposed to toxic compounds in plants. Rumen liquor contains chemical constituents that likely contribute to the beneficial effects of re-establishing a normal reticulo-rumen anaerobic fermentation. Recommendations for collecting rumen fluid, storage and volumes transferred are discussed. Rumen transfaunation is a common practice to treat indigestion on dairy and livestock operations. The support of a healthy microbial community in the digestive tract is also used for humans. Fecal microbiota transplantation has been used to treat digestive disorders in humans. Rumen transfaunation, although not widely studied with respect to mode of action, is an effective, practical, and easy method to treat simple indigestion of ruminants.

  20. Effect of DNA extraction methods and sampling techniques on the apparent structure of cow and sheep rumen microbial communities.

    PubMed

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J; Waghorn, Garry C; Janssen, Peter H

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  1. Effect of DNA Extraction Methods and Sampling Techniques on the Apparent Structure of Cow and Sheep Rumen Microbial Communities

    PubMed Central

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J.; Waghorn, Garry C.; Janssen, Peter H.

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  2. The microbial composition and metabolic potential of the ovine rumen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The rumen is efficient at biotransforming nitroaromatic explosive compounds, such as TNT, RDX, and HMX, which have been used widely in US military munitions. These compounds are present in > 4,000 military items, from large bombs to very small igniters. However, their manufacturing processes have g...

  3. Assessment of the effects of cinnamon leaf oil on rumen microbial fermentation using two continuous culture systems.

    PubMed

    Fraser, G R; Chaves, A V; Wang, Y; McAllister, T A; Beauchemin, K A; Benchaar, C

    2007-05-01

    Two continuous culture (CC) systems, the rumen simulation technique (Rusitec) and a dual-flow (DF) fermenter, were used to evaluate effects of the essential oil from cinnamon leaf (CIN) on rumen microbial fermentation. Incubations (d 1 through 8 for adaptation and d 9 through 16 for sampling) were conducted concurrently in the 2 systems, with CIN added at 0 (control) and 500 mg/L of rumen fluid culture. Eight Rusitec (920 mL; dilution rate = 2.9%/h) and 6 DF (1,300 mL; dilution rate = 6.3%/h) fermenters were randomly assigned to treatment. Inoculum was prepared from 4 ruminally cannulated lactating Holstein cows fed a total mixed ration consisting of 51% forage and 49% concentrate (dry matter basis). Ruminal pH, total volatile fatty acid (VFA) concentration, and diet digestibility were reduced by CIN addition in the Rusitec but were not affected by CIN administration in the DF. The addition of CIN in the Rusitec decreased apparent N disappearance, NH3-N concentration, and molar proportions of branched-chain VFA. In contrast, in the DF no effect of CIN was observed on apparent N degradation, NH3-N concentration, and molar proportion of branched-chain VFA. In the Rusitec, the molar proportion of acetate was similar between treatments on d 9 and 13, but was lower from d 10 to 12 and higher on d 14 to 16 with CIN than with control (interaction of treatment x sampling day). The molar proportion of acetate remained unaffected by CIN addition in the DF. In both CC systems, the molar proportion of propionate was decreased whereas that of butyrate was increased by CIN addition. In the DF, CIN decreased microbial N flow and efficiency of microbial protein synthesis. Protozoa numbers were lower with CIN than with control in both CC fermenters. In the Rusitec, CIN increased 15N enrichment in total bacterial fractions, but no effect was observed on the production of microbial N. This study showed that CIN exhibited antimicrobial activity in both CC systems, but the effects were

  4. Microbial degradation of usnic acid in the reindeer rumen.

    PubMed

    Sundset, Monica A; Barboza, Perry S; Green, Thomas K; Folkow, Lars P; Blix, Arnoldus Schytte; Mathiesen, Svein D

    2010-03-01

    Reindeer (Rangifer tarandus) eat and utilize lichens as an important source of energy and nutrients in winter. Lichens synthesize and accumulate a wide variety of phenolic secondary compounds, such as usnic acid, as a defense against herbivores and to protect against damage by UV-light in solar radiation. We have examined where and to what extent these phenolic compounds are degraded in the digestive tract of the reindeer, with particular focus on usnic acid. Three male reindeer were given ad libitum access to a control diet containing no usnic acid for three weeks and then fed lichens ad libitum (primarily Cladonia stellaris) containing 9.1 mg/g DM usnic acid for 4 weeks. Usnic acid intake in reindeer on the lichen diet was 91-117 mg/kg BM/day. In spite of this, no trace of usnic acid or conjugates of usnic acid was found either in fresh rumen fluid, urine, or feces. This suggests that usnic acid is rapidly degraded by rumen microbes, and that it consequently is not absorbed by the animal. This apparent ability to detoxify lichen phenolic compounds may gain increased importance with future enhanced UV-B radiation expected to cause increased protective usnic acid/phenol production in lichens.

  5. Microbial degradation of usnic acid in the reindeer rumen

    NASA Astrophysics Data System (ADS)

    Sundset, Monica A.; Barboza, Perry S.; Green, Thomas K.; Folkow, Lars P.; Blix, Arnoldus Schytte; Mathiesen, Svein D.

    2010-03-01

    Reindeer ( Rangifer tarandus) eat and utilize lichens as an important source of energy and nutrients in winter. Lichens synthesize and accumulate a wide variety of phenolic secondary compounds, such as usnic acid, as a defense against herbivores and to protect against damage by UV-light in solar radiation. We have examined where and to what extent these phenolic compounds are degraded in the digestive tract of the reindeer, with particular focus on usnic acid. Three male reindeer were given ad libitum access to a control diet containing no usnic acid for three weeks and then fed lichens ad libitum (primarily Cladonia stellaris) containing 9.1 mg/g DM usnic acid for 4 weeks. Usnic acid intake in reindeer on the lichen diet was 91-117 mg/kg BM/day. In spite of this, no trace of usnic acid or conjugates of usnic acid was found either in fresh rumen fluid, urine, or feces. This suggests that usnic acid is rapidly degraded by rumen microbes, and that it consequently is not absorbed by the animal. This apparent ability to detoxify lichen phenolic compounds may gain increased importance with future enhanced UV-B radiation expected to cause increased protective usnic acid/phenol production in lichens.

  6. Buccal swabbing as a noninvasive method to determine bacterial, archaeal, and eukaryotic microbial community structures in the rumen.

    PubMed

    Kittelmann, Sandra; Kirk, Michelle R; Jonker, Arjan; McCulloch, Alan; Janssen, Peter H

    2015-11-01

    Analysis of rumen microbial community structure based on small-subunit rRNA marker genes in metagenomic DNA samples provides important insights into the dominant taxa present in the rumen and allows assessment of community differences between individuals or in response to treatments applied to ruminants. However, natural animal-to-animal variation in rumen microbial community composition can limit the power of a study considerably, especially when only subtle differences are expected between treatment groups. Thus, trials with large numbers of animals may be necessary to overcome this variation. Because ruminants pass large amounts of rumen material to their oral cavities when they chew their cud, oral samples may contain good representations of the rumen microbiota and be useful in lieu of rumen samples to study rumen microbial communities. We compared bacterial, archaeal, and eukaryotic community structures in DNAs extracted from buccal swabs to those in DNAs from samples collected directly from the rumen by use of a stomach tube for sheep on four different diets. After bioinformatic depletion of potential oral taxa from libraries of samples collected via buccal swabs, bacterial communities showed significant clustering by diet (R = 0.37; analysis of similarity [ANOSIM]) rather than by sampling method (R = 0.07). Archaeal, ciliate protozoal, and anaerobic fungal communities also showed significant clustering by diet rather than by sampling method, even without adjustment for potentially orally associated microorganisms. These findings indicate that buccal swabs may in future allow quick and noninvasive sampling for analysis of rumen microbial communities in large numbers of ruminants.

  7. Buccal swabbing as a noninvasive method to determine bacterial, archaeal, and eukaryotic microbial community structures in the rumen.

    PubMed

    Kittelmann, Sandra; Kirk, Michelle R; Jonker, Arjan; McCulloch, Alan; Janssen, Peter H

    2015-11-01

    Analysis of rumen microbial community structure based on small-subunit rRNA marker genes in metagenomic DNA samples provides important insights into the dominant taxa present in the rumen and allows assessment of community differences between individuals or in response to treatments applied to ruminants. However, natural animal-to-animal variation in rumen microbial community composition can limit the power of a study considerably, especially when only subtle differences are expected between treatment groups. Thus, trials with large numbers of animals may be necessary to overcome this variation. Because ruminants pass large amounts of rumen material to their oral cavities when they chew their cud, oral samples may contain good representations of the rumen microbiota and be useful in lieu of rumen samples to study rumen microbial communities. We compared bacterial, archaeal, and eukaryotic community structures in DNAs extracted from buccal swabs to those in DNAs from samples collected directly from the rumen by use of a stomach tube for sheep on four different diets. After bioinformatic depletion of potential oral taxa from libraries of samples collected via buccal swabs, bacterial communities showed significant clustering by diet (R = 0.37; analysis of similarity [ANOSIM]) rather than by sampling method (R = 0.07). Archaeal, ciliate protozoal, and anaerobic fungal communities also showed significant clustering by diet rather than by sampling method, even without adjustment for potentially orally associated microorganisms. These findings indicate that buccal swabs may in future allow quick and noninvasive sampling for analysis of rumen microbial communities in large numbers of ruminants. PMID:26276109

  8. Buccal Swabbing as a Noninvasive Method To Determine Bacterial, Archaeal, and Eukaryotic Microbial Community Structures in the Rumen

    PubMed Central

    Kirk, Michelle R.; Jonker, Arjan; McCulloch, Alan

    2015-01-01

    Analysis of rumen microbial community structure based on small-subunit rRNA marker genes in metagenomic DNA samples provides important insights into the dominant taxa present in the rumen and allows assessment of community differences between individuals or in response to treatments applied to ruminants. However, natural animal-to-animal variation in rumen microbial community composition can limit the power of a study considerably, especially when only subtle differences are expected between treatment groups. Thus, trials with large numbers of animals may be necessary to overcome this variation. Because ruminants pass large amounts of rumen material to their oral cavities when they chew their cud, oral samples may contain good representations of the rumen microbiota and be useful in lieu of rumen samples to study rumen microbial communities. We compared bacterial, archaeal, and eukaryotic community structures in DNAs extracted from buccal swabs to those in DNAs from samples collected directly from the rumen by use of a stomach tube for sheep on four different diets. After bioinformatic depletion of potential oral taxa from libraries of samples collected via buccal swabs, bacterial communities showed significant clustering by diet (R = 0.37; analysis of similarity [ANOSIM]) rather than by sampling method (R = 0.07). Archaeal, ciliate protozoal, and anaerobic fungal communities also showed significant clustering by diet rather than by sampling method, even without adjustment for potentially orally associated microorganisms. These findings indicate that buccal swabs may in future allow quick and noninvasive sampling for analysis of rumen microbial communities in large numbers of ruminants. PMID:26276109

  9. Effect of fumarate reducing bacteria on in vitro rumen fermentation, methane mitigation and microbial diversity.

    PubMed

    Mamuad, Lovelia; Kim, Seon Ho; Jeong, Chang Dae; Choi, Yeon Jae; Jeon, Che Ok; Lee, Sang-Suk

    2014-02-01

    The metabolic pathways involved in hydrogen (H(2)) production, utilization and the activity of methanogens are the important factors that should be considered in controlling methane (CH(4)) emissions by ruminants. H(2) as one of the major substrate for CH(4) production is therefore should be controlled. One of the strategies on reducing CH(4) is through the use of hydrogenotrophic microorganisms such as fumarate reducing bacteria. This study determined the effect of fumarate reducing bacteria, Mitsuokella jalaludinii, supplementation on in vitro rumen fermentation, CH(4) production, diversity and quantity. M. jalaludinii significantly reduced CH(4) at 48 and 72 h of incubation and significantly increased succinate at 24 h. Although not significantly different, propionate was found to be highest in treatment containing M. jalaludinii at 12 and 48 h of incubation. These results suggest that supplementation of fumarate reducing bacteria to ruminal fermentation reduces CH(4) production and quantity, increases succinate and changes the rumen microbial diversity.

  10. Comparison of rumen microbial inhibition resulting from various essential oils isolated from relatively unpalatable plant species.

    PubMed

    Oh, H K; Jones, M B; Longhurst, W M

    1968-01-01

    Essential oils were isolated from eight plant species which were relatively unpalatable to sheep and deer. The inhibitory potency of these essential oils upon sheep and deer rumen microorganisms was compared, in terms of total gas and volatile fatty acid (VFA) production, by use of an anaerobic manometric technique. Inhibitory effects of oils from the eight plant species may be placed in four groups: (i) essential oils from vinegar weed (Trichostema lanceoletum) and California bay (Umbellularia californica) inhibited rumen microbial activity most; (ii) lesser inhibition was exhibited by rosemary (Rosmarinus officinalis) and California mugwort (Artemisia douglasiana) oils, followed by (iii) blue-gum eucalyptus (Eucalyptus globulus) and sagebrush (Artemisia tridentata) oils; and (iv) oils from Douglas fir (Psuedotsuga menziesii) and Jerusalem oak (chenopodium botrys) resulted in the least inhibition, when 0.3 ml of each oil was used. A highly significant correlation coefficient (r = 0.98(**)) between total gas and VFA production indicated the validity of either method to measure the activity of rumen microorganisms. Our results are discussed in relation to the hypothesis that the selectivity and voluntary consumption of ruminants are related to the characteristic odor and antibacterial action of essential oils isolated from relatively unpalatable plant species. PMID:5636470

  11. Effect of different levels of phosphorus on rumen microbial fermentation and synthesis determined using a continuous culture technique.

    PubMed

    Komisarczuk, S; Merry, R J; McAllan, A B

    1987-03-01

    A continuous culture technique was used to study the phosphorus requirements of rumen micro-organisms. Solutions of artificial saliva containing 120, 80, 40 and 0 mg inorganic phosphorus (Pi)/l were infused into the reaction vessels previously inoculated with rumen contents, resulting in Pi concentrations in the vessel contents of 48, 28, 4 and less than 1 mg/l respectively. Various fermentative and synthetic characteristics were examined. In the vessel contents, concentrations of protozoa (about 0.9 X 10(5)/ml) were not significantly affected by Pi concentration. Total volatile fatty acids (VFA) produced averaged about 6.83 mmol/h with Pi levels of 48 and 28 mg/l. Reduction in Pi concentrations to 4 and less than 1 mg/l resulted in significant reductions in total VFA to approximately 6.25 and 3.75 mmol/h respectively, accompanied by a rise in pH from 6.5 to 7.3. Ammonia-nitrogen values, which averaged about 131 mg/l at the higher Pi concentrations, also increased with the lowest level of Pi to about 240 mg/l. ATP concentrations averaged about 14 mumol/l at the highest Pi concentration and fell progressively with each reduction in Pi concentration to a final value of 2.5 mumol/l with the Pi level less than 1 mg/l. At Pi concentrations of 48 and 28 mg/l, the digestibilities of xylose, arabinose and cellulose-glucose were maintained at about 0.90, 0.62 and 0.70 g/g input respectively. At lower Pi concentrations these digestibilities fell significantly and corresponding values at Pi less than 1 mg/l were 0.73, 0.41 and 0.31 respectively. Starch digestion was unaffected by Pi concentration and remained at about 0.90 g/g input. The amount of microbial-N synthesized averaged 0.48 g/d and was maintained with Pi concentrations down to 4 mg/l. There was, however, a significant reduction to 0.26 g/d with Pi concentrations of less than 1 mg/l. The efficiency of microbial protein synthesis was variable but averaged approximately 25 g N/kg total carbohydrate fermented. It was

  12. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community

    PubMed Central

    Tapio, Ilma; Shingfield, Kevin J.; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R.; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J.; Wallace, R. John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint. PMID:26986467

  13. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community.

    PubMed

    Tapio, Ilma; Shingfield, Kevin J; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J; Wallace, R John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint. PMID:26986467

  14. Microbial biodiversity of the liquid fraction of rumen content from lactating cows.

    PubMed

    Sandri, M; Manfrin, C; Pallavicini, A; Stefanon, B

    2014-04-01

    Host and dietary interactions with the rumen microbiome can affect the efficacy of supplements, and their effect on the composition of the bacterial population is still unknown. A 16S rRNA metagenomic approach and Next-Generation Sequencing (NGS) technology were used to investigate the bacterial microbiome composition in the liquid fraction of the rumen content collected via stomach tubing. To investigate biodiversity, samples were taken from three groups of four lactating dairy cows given a supplement of either 50 g of potato protein (Ctrl group), or 50 g of lyophilized Saccharomyces cerevisiae (LY group) or 50 g of dried S. cerevisiae (DY group) in a potato protein support. Rumen samples were collected after 15 days of dietary treatments and milk production was similar between the three groups. Taxonomic distribution analysis revealed a prevalence of the Firmicutes phylum in all cows (79.76%) and a significantly (P<0.05) higher presence of the genus Bacillus in the DY group. Volatile fatty-acid concentration was not significantly different between groups, possibly because of relatively high inter-animal variability or limited effect of the treatments or both, and the correlation analysis with bacterial taxa showed significant associations, in particular between many Firmicutes genera and butyrate. Limited differences were observed between dietary treatments, but the lack of microbiome data before yeast administration does not allow to draw firm conclusions on the effect of dietary treatments. PMID:24524278

  15. Metagenomic analysis of the rumen microbial community following inhibition of methane formation by a halogenated methane analog.

    PubMed

    Denman, Stuart E; Martinez Fernandez, Gonzalo; Shinkai, Takumi; Mitsumori, Makoto; McSweeney, Christopher S

    2015-01-01

    Japanese goats fed a diet of 50% Timothy grass and 50% concentrate with increasing levels of the anti-methanogenic compound, bromochloromethane (BCM) were investigated with respect to the microbial population and functional shifts in the rumen. Microbial ecology methods identified species that exhibited positive and negative responses to the increasing levels of BCM. The methane-inhibited rumen appeared to adapt to the higher H2 levels by shifting fermentation to propionate which was mediated by an increase in the population of H2-consuming Prevotella and Selenomonas spp. Metagenomic analysis of propionate production pathways was dominated by genomic content from these species. Reductive acetogenic marker gene libraries and metagenomics analysis indicate that reductive acetogenic species do not play a major role in the BCM treated rumen.

  16. Metagenomic analysis of the rumen microbial community following inhibition of methane formation by a halogenated methane analog

    PubMed Central

    Denman, Stuart E.; Martinez Fernandez, Gonzalo; Shinkai, Takumi; Mitsumori, Makoto; McSweeney, Christopher S.

    2015-01-01

    Japanese goats fed a diet of 50% Timothy grass and 50% concentrate with increasing levels of the anti-methanogenic compound, bromochloromethane (BCM) were investigated with respect to the microbial population and functional shifts in the rumen. Microbial ecology methods identified species that exhibited positive and negative responses to the increasing levels of BCM. The methane-inhibited rumen appeared to adapt to the higher H2 levels by shifting fermentation to propionate which was mediated by an increase in the population of H2-consuming Prevotella and Selenomonas spp. Metagenomic analysis of propionate production pathways was dominated by genomic content from these species. Reductive acetogenic marker gene libraries and metagenomics analysis indicate that reductive acetogenic species do not play a major role in the BCM treated rumen. PMID:26528253

  17. Metabolism of soluble rapeseed meal (Brassica rapa L.) protein during incubations with buffered bovine rumen contents in vitro.

    PubMed

    Stefański, T; Ahvenjärvi, S; Huhtanen, P; Shingfield, K J

    2013-01-01

    Accurate quantitative information on the fate of dietary protein in the rumen is central to modern metabolizable protein systems developed to improve the efficiency of nitrogen utilization in ruminants. An in vitro method was developed to estimate the rate of soluble rapeseed meal (Brassica rapa L.) protein (SRMP) degradation. Unlabeled and (15)N-labeled solvent-extracted rapeseed meal were incubated alone or as an equal mixture (125 mg of N/L) with buffered rumen contents and a mixture of carbohydrates formulated to provide a constant source of fermentable energy during the course of all incubations. Incubations were made over 0.33, 0.67, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 6.0, 8.0, and 10.0 h. Enrichment of (14)N and (15)N isotopes in total N of ammonia (AN), soluble nonammonia (SNAN), and insoluble (ISN) fractions liberated during incubations with test proteins was determined. A model with 4 pools that accounted for both intracellular and extracellular N transformations was used to estimate the rate of SRMP degradation. Parameter values used in the model were adjusted based on the size of A(14)N, A(15)N, SNA(14)N, SNA(15)N, IS(14)N, and IS(15)N pools, measured at different time points during incubations with buffered rumen fluid. The mean rate of N degradation for SRMP was estimated at 0.126 (SD 0.0499) h(-1). No substantive difference in the rate of protein degradation or microbial protein synthesis was observed during incubations of labeled and unlabeled substrates with rumen fluid. In conclusion, combined use of data from incubations of unlabeled and (15)N-labeled rapeseed protein with buffered rumen inoculum provided sufficient information to allow for estimation of parameter values in a complex dynamic model of soluble protein degradation. Results indicate the potential of the technique to evaluate the degradability of SNAN of other dietary protein sources and implicate ruminal escape of soluble rapeseed protein as an important source of amino acids in ruminants.

  18. Microbial and Carbohydrate Active Enzyme profile of buffalo rumen metagenome and their alteration in response to variation in the diet.

    PubMed

    Patel, Dishita D; Patel, Amrutlal K; Parmar, Nidhi R; Shah, Tejas M; Patel, Jethabhai B; Pandya, Paresh R; Joshi, Chaitanya G

    2014-07-15

    Rumen microbiome represents rich source of enzymes degrading complex plant polysaccharides. We describe here analysis of Carbohydrate Active Enzymes (CAZymes) from 3.5 gigabase sequences of metagenomic data from rumen samples of Mehsani buffaloes fed on different proportions of green or dry roughages to concentrate ration. A total of 2597 contigs encoding putative CAZymes were identified by CAZyme Analysis Toolkit (CAT). The phylogenetic analysis of these contigs by MG-RAST revealed predominance of Bacteroidetes, followed by Firmicutes, Proteobacteria, and Actinobacteria phyla. Moreover, a higher abundance of oligosaccharide degrading and debranching enzymes in buffalo rumen metagenome and that of cellulases and hemicellulases in termite hindgut was observed when we compared glycoside hydrolase (GH) profile of buffalo rumen metagenome with cow rumen, termite hindgut and chicken caecum metagenome. Further, comparison of microbial profile of green or dry roughage fed animals showed significantly higher abundance (p-value<0.05) of various polysaccharide degrading bacterial genera like Fibrobacter, Prevotella, Bacteroides, Clostridium and Ruminococcus in green roughage fed animals. In addition, we found a significantly higher abundance (p-value<0.05) of enzymes associated with pectin digestion such as pectin lyase (PL) 1, PL10 and GH28 in green roughage fed animals. Our study outlines CAZyme profile of buffalo rumen metagenome and provides a scope to study the role of abundant enzyme families (oligosaccharide degrading and debranching enzymes) in digestion of coarse feed. PMID:24797613

  19. Manipulation of soluble and rumen-undegradable protein in diets fed to postpubertal dairy heifers.

    PubMed

    Zanton, G I; Gabler, M T; Heinrichs, A J

    2007-02-01

    Eight postpubertal Holstein heifers (455 +/- 4.0), fit with rumen cannulas, were used in 2 experiments to investigate the effects of altering dietary protein type on nutrient digestibility, rumen fermentation, and nitrogen utilization. Heifers were fed diets containing low or high levels of soluble (SP) and low or high levels of rumen-undegradable protein (RUP) in a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. The treatment rations in experiment 1 were formulated with corn silage composing the majority of the forage fraction, whereas in experiment 2, grass hay composed the highest proportion of ration DM. Blood and rumen samples were collected over 2 d and total fecal and urine collections were conducted for 4 d. Dry matter, organic matter, and neutral detergent fiber digestibility were not different in either experiment 1 or 2. Increasing the proportion of dietary crude protein that was SP increased mean daily rumen ammonia concentrations in each experiment, although no other rumen parameter differed. Excretion of urinary nitrogen in experiment 1 was highest for diets with low SP and low RUP and with high SP and high RUP, which resulted in these rations being the least efficient in retention of apparently digested nitrogen. The proportion of consumed or absorbed nitrogen retained in experiment 2 was not significantly different between treatments. Responses to alterations in crude protein degradability are observable in postpubertal heifers; however, the level of response may depend on the diet in which protein degradability is altered. PMID:17235175

  20. Effect of direct-fed microbials on performance, diet digestibility, and rumen characteristics of Holstein dairy cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to determine the effect of feeding bacterial direct fed microbial (DFM) products containing Lactobacillus acidophilus (LA) and Propionibacteria freudenreichii (PF) on the performance, nutrient digestibility, and rumen fermentation of Holstein dairy cows in mid-lactati...

  1. Responses in digestion, rumen fermentation and microbial populations to inhibition of methane formation by a halogenated methane analogue.

    PubMed

    Mitsumori, Makoto; Shinkai, Takumi; Takenaka, Akio; Enishi, Osamu; Higuchi, Koji; Kobayashi, Yosuke; Nonaka, Itoko; Asanuma, Narito; Denman, Stuart E; McSweeney, Christopher S

    2012-08-01

    The effects of the anti-methanogenic compound, bromochloromethane (BCM), on rumen microbial fermentation and ecology were examined in vivo. Japanese goats were fed a diet of 50 % Timothy grass and 50 % concentrate and then sequentially adapted to low, mid and high doses of BCM. The goats were placed into the respiration chambers for analysis of rumen microbial function and methane and H2 production. The levels of methane production were reduced by 5, 71 and 91 %, and H2 production was estimated at 545, 2941 and 3496 mmol/head per d, in response to low, mid and high doses of BCM, respectively, with no effect on maintenance feed intake and digestibility. Real-time PCR quantification of microbial groups showed a significant decrease relative to controls in abundance of methanogens and rumen fungi, whereas there were increases in Prevotella spp. and Fibrobacter succinogenes, a decrease in Ruminococcus albus and R. flavefaciens was unchanged. The numbers of protozoa were also unaffected. Denaturing gradient gel electrophoresis and quantitative PCR analysis revealed that several Prevotella spp. were the bacteria that increased most in response to BCM treatment. It is concluded that the methane-inhibited rumen adapts to high hydrogen levels by shifting fermentation to propionate via Prevotella spp., but the majority of metabolic hydrogen is expelled as H2 gas.

  2. Rumen Degradability and Post-ruminal Digestion of Dry Matter, Nitrogen and Amino Acids of Three Protein Supplements

    PubMed Central

    Gao, Wei; Chen, Aodong; Zhang, Bowen; Kong, Ping; Liu, Chenli; Zhao, Jie

    2015-01-01

    This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID) of dry matter, crude protein (CP), and amino acids (AA) of cottonseed meal (CSM), sunflower seed meal (SFSM) and distillers dried grains with solubles (DDGS) by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA) varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP) content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen micro-organisms, especially for Trp, Arg, His, and Lys (p<0.01). The SID of individual RUP-AA was different within specific RUP origin (p<0.01). Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance. PMID:25656208

  3. Rumen microbial abundance and fermentation profile during severe subacute ruminal acidosis and its modulation by plant derived alkaloids in vitro.

    PubMed

    Mickdam, Elsayed; Khiaosa-Ard, Ratchaneewan; Metzler-Zebeli, Barbara U; Klevenhusen, Fenja; Chizzola, Remigius; Zebeli, Qendrim

    2016-06-01

    Rumen microbiota have important metabolic functions for the host animal. This study aimed at characterizing changes in rumen microbial abundances and fermentation profiles using a severe subacute ruminal acidosis (SARA) in vitro model, and to evaluate a potential modulatory role of plant derived alkaloids (PDA), containing quaternary benzophenanthridine and protopine alkaloids, of which sanguinarine and chelerythrine were the major bioactive compounds. Induction of severe SARA strongly affected the rumen microbial composition and fermentation variables without suppressing the abundance of total bacteria. Protozoa and fungi were more sensitive to the low ruminal pH condition than bacteria. Induction of severe SARA clearly depressed degradation of fiber (P < 0.001), which came along with a decreased relative abundance of fibrolytic Ruminococcus albus and Fibrobacter succinogenes (P < 0.001). Under severe SARA conditions, the genus Prevotella, Lactobacillus group, Megasphaera elsdenii, and Entodinium spp. (P < 0.001) were more abundant, whereas Ruminobacter amylophilus was less abundant. SARA largely suppressed methane formation (-70%, P < 0.001), although total methanogenic 16S rRNA gene abundance was not affected. According to principal component analysis, Methanobrevibacter spp. correlated to methane concentration. Addition of PDA modulated ruminal fermentation under normal conditions such as enhanced (P < 0.05) concentration of total SCFA, propionate and valerate, and increased (P < 0.05) degradation of crude protein compared with the unsupplemented control diet. Our results indicate strong shifts in the microbial community during severe SARA compared to normal conditions. Supplementation of PDA positively modulates ruminal fermentation under normal ruminal pH conditions. PMID:26868619

  4. Effect of phenotypic residual feed intake and dietary forage content on the rumen microbial community of beef cattle.

    PubMed

    Carberry, Ciara A; Kenny, David A; Han, Sukkyan; McCabe, Matthew S; Waters, Sinead M

    2012-07-01

    Feed-efficient animals have lower production costs and reduced environmental impact. Given that rumen microbial fermentation plays a pivotal role in host nutrition, the premise that rumen microbiota may contribute to host feed efficiency is gaining momentum. Since diet is a major factor in determining rumen community structure and fermentation patterns, we investigated the effect of divergence in phenotypic residual feed intake (RFI) on ruminal community structure of beef cattle across two contrasting diets. PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR) were performed to profile the rumen bacterial population and to quantify the ruminal populations of Entodinium spp., protozoa, Fibrobacter succinogenes, Ruminococcus flavefaciens, Ruminococcus albus, Prevotella brevis, the genus Prevotella, and fungi in 14 low (efficient)- and 14 high (inefficient)-RFI animals offered a low-energy, high-forage diet, followed by a high-energy, low-forage diet. Canonical correspondence and Spearman correlation analyses were used to investigate associations between physiological variables and rumen microbial structure and specific microbial populations, respectively. The effect of RFI on bacterial profiles was influenced by diet, with the association between RFI group and PCR-DGGE profiles stronger for the higher forage diet. qPCR showed that Prevotella abundance was higher (P < 0.0001) in inefficient animals. A higher (P < 0.0001) abundance of Entodinium and Prevotella spp. and a lower (P < 0.0001) abundance of Fibrobacter succinogenes were observed when animals were offered the low-forage diet. Thus, differences in the ruminal microflora may contribute to host feed efficiency, although this effect may also be modulated by the diet offered.

  5. Microbial population in the rumen of swamp buffalo (Bubalus bubalis) as influenced by coconut oil and mangosteen peel supplementation.

    PubMed

    Pilajun, R; Wanapat, M

    2013-06-01

    Four, rumen fistulated swamp buffalo bulls were used to study microbial populations in the rumen when supplemented with coconut oil and mangosteen peel. Animals were randomly assigned to a 4 × 4 Latin square design. Four treatments were un-supplemented (Control), supplementation with coconut oil at 50 g/kg (CO5), supplementation with mangosteen peel at 30 g/kg (MP3) and supplementation with CO5 and MP3 (COM), of total DM intake. Animals received concentrate at 10 g/kg of BW, and rice straw was given ad libitum. Abundance of total bacteria was increased by CO5 supplementation, whereas populations of protozoa and Fibrobacter succinogenes were reduced by CO5 and COM supplementation. Dietary supplementation did not affect methanogen, Ruminococcus flavefaciens or Ruminococcus albus abundances. Dietary treatments changed denaturing gradient gel electrophoresis (DGGE) band patterns of methanogens and protozoa when compared with the control group, especially when supplemented with MP3. Supplementation of COM resulted in the greatest difference in pattern of DGGE bands for total bacteria compared with the control. Coconut oil and mangosteen peel supplementation resulted in changing of rumen microbial abundances and communities; however, combination of them could be more benefit to improve rumen fermentation of swamp buffalo fed on rice straw.

  6. Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range.

    PubMed

    Henderson, Gemma; Cox, Faith; Ganesh, Siva; Jonker, Arjan; Young, Wayne; Janssen, Peter H

    2015-01-01

    Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific.

  7. Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range

    PubMed Central

    Henderson, Gemma; Cox, Faith; Ganesh, Siva; Jonker, Arjan; Young, Wayne; Abecia, Leticia; Angarita, Erika; Aravena, Paula; Nora Arenas, Graciela; Ariza, Claudia; Attwood, Graeme T.; Mauricio Avila, Jose; Avila-Stagno, Jorge; Bannink, André; Barahona, Rolando; Batistotti, Mariano; Bertelsen, Mads F.; Brown-Kav, Aya; Carvajal, Andres M.; Cersosimo, Laura; Vieira Chaves, Alexandre; Church, John; Clipson, Nicholas; Cobos-Peralta, Mario A.; Cookson, Adrian L.; Cravero, Silvio; Cristobal Carballo, Omar; Crosley, Katie; Cruz, Gustavo; Cerón Cucchi, María; de la Barra, Rodrigo; De Menezes, Alexandre B.; Detmann, Edenio; Dieho, Kasper; Dijkstra, Jan; dos Reis, William L. S.; Dugan, Mike E. R.; Hadi Ebrahimi, Seyed; Eythórsdóttir, Emma; Nde Fon, Fabian; Fraga, Martín; Franco, Francisco; Friedeman, Chris; Fukuma, Naoki; Gagić, Dragana; Gangnat, Isabelle; Javier Grilli, Diego; Guan, Le Luo; Heidarian Miri, Vahideh; Hernandez-Sanabria, Emma; Gomez, Alma Ximena Ibarra; Isah, Olubukola A.; Ishaq, Suzanne; Jami, Elie; Jelincic, Juan; Kantanen, Juha; Kelly, William J.; Kim, Seon-Ho; Klieve, Athol; Kobayashi, Yasuo; Koike, Satoshi; Kopecny, Jan; Nygaard Kristensen, Torsten; Julie Krizsan, Sophie; LaChance, Hannah; Lachman, Medora; Lamberson, William R.; Lambie, Suzanne; Lassen, Jan; Leahy, Sinead C.; Lee, Sang-Suk; Leiber, Florian; Lewis, Eva; Lin, Bo; Lira, Raúl; Lund, Peter; Macipe, Edgar; Mamuad, Lovelia L.; Cuquetto Mantovani, Hilário; Marcoppido, Gisela Ariana; Márquez, Cristian; Martin, Cécile; Martinez, Gonzalo; Eugenia Martinez, Maria; Lucía Mayorga, Olga; McAllister, Tim A.; McSweeney, Chris; Mestre, Lorena; Minnee, Elena; Mitsumori, Makoto; Mizrahi, Itzhak; Molina, Isabel; Muenger, Andreas; Munoz, Camila; Murovec, Bostjan; Newbold, John; Nsereko, Victor; O’Donovan, Michael; Okunade, Sunday; O’Neill, Brendan; Ospina, Sonia; Ouwerkerk, Diane; Parra, Diana; Pereira, Luiz Gustavo Ribeiro; Pinares-Patino, Cesar; Pope, Phil B.; Poulsen, Morten; Rodehutscord, Markus; Rodriguez, Tatiana; Saito, Kunihiko; Sales, Francisco; Sauer, Catherine; Shingfield, Kevin; Shoji, Noriaki; Simunek, Jiri; Stojanović-Radić, Zorica; Stres, Blaz; Sun, Xuezhao; Swartz, Jeffery; Liang Tan, Zhi; Tapio, Ilma; Taxis, Tasia M.; Tomkins, Nigel; Ungerfeld, Emilio; Valizadeh, Reza; van Adrichem, Peter; Van Hamme, Jonathan; Van Hoven, Woulter; Waghorn, Garry; John Wallace, R.; Wang, Min; Waters, Sinéad M.; Keogh, Kate; Witzig, Maren; Wright, Andre-Denis G.; Yamano, Hidehisa; Yan, Tianhai; Yanez-Ruiz, David R.; Yeoman, Carl J.; Zambrano, Ricardo; Zeitz, Johanna; Zhou, Mi; Wei Zhou, Hua; Xia Zou, Cai; Zunino, Pablo; Janssen, Peter H.

    2015-01-01

    Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific. PMID:26449758

  8. Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range.

    PubMed

    Henderson, Gemma; Cox, Faith; Ganesh, Siva; Jonker, Arjan; Young, Wayne; Janssen, Peter H

    2015-01-01

    Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific. PMID:26449758

  9. Simultaneous Amplicon Sequencing to Explore Co-Occurrence Patterns of Bacterial, Archaeal and Eukaryotic Microorganisms in Rumen Microbial Communities

    PubMed Central

    Kittelmann, Sandra; Seedorf, Henning; Walters, William A.; Clemente, Jose C.; Knight, Rob; Gordon, Jeffrey I.; Janssen, Peter H.

    2013-01-01

    Ruminants rely on a complex rumen microbial community to convert dietary plant material to energy-yielding products. Here we developed a method to simultaneously analyze the community's bacterial and archaeal 16S rRNA genes, ciliate 18S rRNA genes and anaerobic fungal internal transcribed spacer 1 genes using 12 DNA samples derived from 11 different rumen samples from three host species (Ovis aries, Bos taurus, Cervus elephas) and multiplex 454 Titanium pyrosequencing. We show that the mixing ratio of the group-specific DNA templates before emulsion PCR is crucial to compensate for differences in amplicon length. This method, in contrast to using a non-specific universal primer pair, avoids sequencing non-targeted DNA, such as plant- or endophyte-derived rRNA genes, and allows increased or decreased levels of community structure resolution for each microbial group as needed. Communities analyzed with different primers always grouped by sample origin rather than by the primers used. However, primer choice had a greater impact on apparent archaeal community structure than on bacterial community structure, and biases for certain methanogen groups were detected. Co-occurrence analysis of microbial taxa from all three domains of life suggested strong within- and between-domain correlations between different groups of microorganisms within the rumen. The approach used to simultaneously characterize bacterial, archaeal and eukaryotic components of a microbiota should be applicable to other communities occupying diverse habitats. PMID:23408926

  10. Simultaneous amplicon sequencing to explore co-occurrence patterns of bacterial, archaeal and eukaryotic microorganisms in rumen microbial communities.

    PubMed

    Kittelmann, Sandra; Seedorf, Henning; Walters, William A; Clemente, Jose C; Knight, Rob; Gordon, Jeffrey I; Janssen, Peter H

    2013-01-01

    Ruminants rely on a complex rumen microbial community to convert dietary plant material to energy-yielding products. Here we developed a method to simultaneously analyze the community's bacterial and archaeal 16S rRNA genes, ciliate 18S rRNA genes and anaerobic fungal internal transcribed spacer 1 genes using 12 DNA samples derived from 11 different rumen samples from three host species (Ovis aries, Bos taurus, Cervus elephas) and multiplex 454 Titanium pyrosequencing. We show that the mixing ratio of the group-specific DNA templates before emulsion PCR is crucial to compensate for differences in amplicon length. This method, in contrast to using a non-specific universal primer pair, avoids sequencing non-targeted DNA, such as plant- or endophyte-derived rRNA genes, and allows increased or decreased levels of community structure resolution for each microbial group as needed. Communities analyzed with different primers always grouped by sample origin rather than by the primers used. However, primer choice had a greater impact on apparent archaeal community structure than on bacterial community structure, and biases for certain methanogen groups were detected. Co-occurrence analysis of microbial taxa from all three domains of life suggested strong within- and between-domain correlations between different groups of microorganisms within the rumen. The approach used to simultaneously characterize bacterial, archaeal and eukaryotic components of a microbiota should be applicable to other communities occupying diverse habitats.

  11. Description of the structural diversity of rumen microbial communities in vitro using single-strand conformation polymorphism profiles.

    PubMed

    Boguhn, Jeannette; Strobel, Egbert; Witzig, Maren; Tebbe, Christoph C; Rodehutscord, Markus

    2008-12-01

    Changes of the rumen microbial community structure, as it can be established with a rumen simulation technique (RUSITEC) were studied using PCR and single-strand conformation polymorphism (SSCP) of small subunit rDNA genes (SSU rDNA). Four total mixed rations were incubated and two ammonia levels in the artificial saliva were applied. Three replicated vessels were used for each treatment. Mixed microbial fractions were isolated by stepwise centrifugation from the liquid fraction (reference microbes, RM) and from the solids of the feed residues (solid-associated microbes, SAM). PCR-primers targeting archaea, fibrobacter, clostridia, and bacteria, respectively, were applied to represent the individual taxonomic groups by SSCP profiles. These SSCP profiles were converted into a binary matrix and distances among treatments were visualised by non-metric multidimensional scaling. Between replicates belonging to one treatment only small differences were found, indicating a high reproducibility of the RUSITEC and the chosen SSCP method. The ammonia concentration seems to be affecting the SSCP profiles. Great differences occurred between RM and SAM, especially for profiles targeting bacteria and clostridia. Differences in the profiles of RM were also found between mixed rations that contained the same feedstuffs in different ratios and between rations with similar nutrient content but based on different feedstuffs. In conclusion, the PCR-SSCP-based technique in conjunction with non-metric multidimensional scaling was sufficiently sensitive to detect and compare changes in composition of rumen microbial community structure in vitro as affected by diet and other environmental factors.

  12. The Role of Ciliate Protozoa in the Rumen

    PubMed Central

    Newbold, Charles J.; de la Fuente, Gabriel; Belanche, Alejandro; Ramos-Morales, Eva; McEwan, Neil R.

    2015-01-01

    First described in 1843, Rumen protozoa with their striking appearance were assumed to be important for the welfare of their host. However, despite contributing up to 50% of the bio-mass in the rumen, the role of protozoa in rumen microbial ecosystem remains unclear. Phylogenetic analysis of 18S rDNA libraries generated from the rumen of cattle, sheep, and goats has revealed an unexpected diversity of ciliated protozoa although variation in gene copy number between species makes it difficult to obtain absolute quantification. Despite repeated attempts it has proven impossible to maintain rumen protozoa in axenic culture. Thus it has been difficult to establish conclusively a role of ciliate protozoa in rumen fiber degradation. The development of techniques to clone and express ciliate genes in λ phage, together with bioinformatic indices to confirm the ciliate origin of the genes has allowed the isolation and characterization of fibrolytic genes from rumen protozoa. Elimination of the ciliate protozoa increases microbial protein supply by up to 30% and reduces methane production by up to 11%. Our recent findings suggest that holotrich protozoa play a disproportionate role in supporting methanogenesis whilst the small Entodinium are responsible for much of the bacterial protein turnover. As yet no method to control protozoa in the rumen that is safe and practically applicable has been developed, however a range of plant extract capable of controlling if not completely eliminating rumen protozoa have been described. PMID:26635774

  13. The Role of Ciliate Protozoa in the Rumen.

    PubMed

    Newbold, Charles J; de la Fuente, Gabriel; Belanche, Alejandro; Ramos-Morales, Eva; McEwan, Neil R

    2015-01-01

    First described in 1843, Rumen protozoa with their striking appearance were assumed to be important for the welfare of their host. However, despite contributing up to 50% of the bio-mass in the rumen, the role of protozoa in rumen microbial ecosystem remains unclear. Phylogenetic analysis of 18S rDNA libraries generated from the rumen of cattle, sheep, and goats has revealed an unexpected diversity of ciliated protozoa although variation in gene copy number between species makes it difficult to obtain absolute quantification. Despite repeated attempts it has proven impossible to maintain rumen protozoa in axenic culture. Thus it has been difficult to establish conclusively a role of ciliate protozoa in rumen fiber degradation. The development of techniques to clone and express ciliate genes in λ phage, together with bioinformatic indices to confirm the ciliate origin of the genes has allowed the isolation and characterization of fibrolytic genes from rumen protozoa. Elimination of the ciliate protozoa increases microbial protein supply by up to 30% and reduces methane production by up to 11%. Our recent findings suggest that holotrich protozoa play a disproportionate role in supporting methanogenesis whilst the small Entodinium are responsible for much of the bacterial protein turnover. As yet no method to control protozoa in the rumen that is safe and practically applicable has been developed, however a range of plant extract capable of controlling if not completely eliminating rumen protozoa have been described.

  14. The Role of Ciliate Protozoa in the Rumen.

    PubMed

    Newbold, Charles J; de la Fuente, Gabriel; Belanche, Alejandro; Ramos-Morales, Eva; McEwan, Neil R

    2015-01-01

    First described in 1843, Rumen protozoa with their striking appearance were assumed to be important for the welfare of their host. However, despite contributing up to 50% of the bio-mass in the rumen, the role of protozoa in rumen microbial ecosystem remains unclear. Phylogenetic analysis of 18S rDNA libraries generated from the rumen of cattle, sheep, and goats has revealed an unexpected diversity of ciliated protozoa although variation in gene copy number between species makes it difficult to obtain absolute quantification. Despite repeated attempts it has proven impossible to maintain rumen protozoa in axenic culture. Thus it has been difficult to establish conclusively a role of ciliate protozoa in rumen fiber degradation. The development of techniques to clone and express ciliate genes in λ phage, together with bioinformatic indices to confirm the ciliate origin of the genes has allowed the isolation and characterization of fibrolytic genes from rumen protozoa. Elimination of the ciliate protozoa increases microbial protein supply by up to 30% and reduces methane production by up to 11%. Our recent findings suggest that holotrich protozoa play a disproportionate role in supporting methanogenesis whilst the small Entodinium are responsible for much of the bacterial protein turnover. As yet no method to control protozoa in the rumen that is safe and practically applicable has been developed, however a range of plant extract capable of controlling if not completely eliminating rumen protozoa have been described. PMID:26635774

  15. Progress in the development of vaccines against rumen methanogens.

    PubMed

    Wedlock, D N; Janssen, P H; Leahy, S C; Shu, D; Buddle, B M

    2013-06-01

    Vaccination against rumen methanogens offers a practical approach to reduce methane emissions in livestock, particularly ruminants grazing on pasture. Although successful vaccination strategies have been reported for reducing the activity of the rumen-dwelling organism Streptococcus bovis in sheep and S. bovis and Lactobacillus spp. in cattle, earlier approaches using vaccines based on whole methanogen cells to reduce methane production in sheep have produced less promising results. An anti-methanogen vaccine will need to have broad specificity against methanogens commonly found in the rumen and induce antibody in saliva resulting in delivery of sufficiently high levels of antibodies to the rumen to reduce methanogen activity. Our approach has focussed on identifying surface and membrane-associated proteins that are conserved across a range of rumen methanogens. The identification of potential vaccine antigens has been assisted by recent advances in the knowledge of rumen methanogen genomes. Methanogen surface proteins have been shown to be immunogenic in ruminants and vaccination of sheep with these proteins induced specific antibody responses in saliva and rumen contents. Current studies are directed towards identifying key candidate antigens and investigating the level and types of salivary antibodies produced in sheep and cattle vaccinated with methanogen proteins, stability of antibodies in the rumen and their impact on rumen microbial populations. In addition, there is a need to identify adjuvants that stimulate high levels of salivary antibody and are suitable for formulating with protein antigens to produce a low-cost and effective vaccine.

  16. Effects of minerals on feed degradation and protein synthesis by rumen micro-organisms in a dual effluent fermenter.

    PubMed

    Broudiscou, L P; Papon, Y; Broudiscou, A F

    1999-01-01

    In dual outflow continuous fermenters on a 75:25 hay/barley diet, feed degradation and protein synthesis by mixed rumen microbes were tested in relation to the concentrations of HPO4(2-), HCO3- and Cl- and Na+/K+ ratio in artificial saliva, by applying a 16-run Franquart design, and by fitting second-order polynomial models. The HPO4(2-), HCO3-, Cl- concentrations and Na+/K+ ratio ranged from 0.1 to 4 g.L-1, from 0.5 to 7 g.L-1, from 0.1 to 0.5 g.L-1 and from 0.5 to 15 g.g-1, respectively. Buffer salts, particularly HPO4(2-), were the major factors while Cl- concentration had negligible effects on microbial metabolism. Maximal neutral detergent fibre, acid detergent fibre and organic matter degradabilities occurred at intermediate values of HPO4(2-) and HCO3- concentrations. The outflow of microbial protein and the efficiency of microbial protein synthesis, which varied from 26.2 to 37.1 g.N.kg-1 of organic matter truly degraded, reached minima at the centre of the experimental domain. PMID:10327453

  17. Effects of minerals on feed degradation and protein synthesis by rumen micro-organisms in a dual effluent fermenter.

    PubMed

    Broudiscou, L P; Papon, Y; Broudiscou, A F

    1999-01-01

    In dual outflow continuous fermenters on a 75:25 hay/barley diet, feed degradation and protein synthesis by mixed rumen microbes were tested in relation to the concentrations of HPO4(2-), HCO3- and Cl- and Na+/K+ ratio in artificial saliva, by applying a 16-run Franquart design, and by fitting second-order polynomial models. The HPO4(2-), HCO3-, Cl- concentrations and Na+/K+ ratio ranged from 0.1 to 4 g.L-1, from 0.5 to 7 g.L-1, from 0.1 to 0.5 g.L-1 and from 0.5 to 15 g.g-1, respectively. Buffer salts, particularly HPO4(2-), were the major factors while Cl- concentration had negligible effects on microbial metabolism. Maximal neutral detergent fibre, acid detergent fibre and organic matter degradabilities occurred at intermediate values of HPO4(2-) and HCO3- concentrations. The outflow of microbial protein and the efficiency of microbial protein synthesis, which varied from 26.2 to 37.1 g.N.kg-1 of organic matter truly degraded, reached minima at the centre of the experimental domain.

  18. Analysis of rumen microbial populations in lactating dairy cattle fed diets varying in carbohydrate profiles and Saccharomyces cerevisiae fermentation product.

    PubMed

    Mullins, C R; Mamedova, L K; Carpenter, A J; Ying, Y; Allen, M S; Yoon, I; Bradford, B J

    2013-09-01

    The rumen microbial ecosystem is a critical factor that links diets to bovine physiology and productivity; however, information about dietary effects on microbial populations has generally been limited to small numbers of samples and qualitative assessment. To assess whether consistent shifts in microbial populations occur in response to common dietary manipulations in dairy cattle, samples of rumen contents were collected from 2 studies for analysis by quantitative real-time PCR (qPCR). In one study, lactating Holstein cows (n=8) were fed diets in which a nonforage fiber source replaced an increasing proportion of forages and concentrates in a 4×4 Latin square design, and samples of ruminal digesta were collected at 9-h intervals over 3 d at the end of each period. In the second study, lactating Holstein cows (n=15) were fed diets with or without the inclusion of a Saccharomyces cerevisiae fermentation product (SCFP) in a crossover design. In this study, rumen liquid and solid samples were collected during total rumen evacuations before and after feeding in a 42-h period. In total, 146 samples of ruminal digesta were used for microbial DNA isolation and analysis by qPCR. Validated primer sets were used to quantify total bacterial and anaerobic fungal populations as well as 12 well-studied bacterial taxa. The relative abundance of the target populations was similar to those previously reported. No significant treatment effects were observed for any target population. A significant interaction of treatment and dry matter intake was observed, however, for the abundance of Eubacterium ruminantium. Increasing dry matter intake was associated with a quadratic decrease in E. ruminantium populations in control animals but with a quadratic increase in E.ruminantium populations in cows fed SCFP. Analysis of sample time effects revealed that Fibrobacter succinogenes and fungal populations were more abundant postfeeding, whereas Ruminococcus albus tended to be more abundant

  19. Molecular comparative assessment of the microbial ecosystem in rumen and faeces of goats fed alfalfa hay alone or combined with oats.

    PubMed

    Mohammadzadeh, Hamid; Yáñez-Ruiz, David R; Martínez-Fernandez, Gonzalo; Abecia, Leticia

    2014-10-01

    The objective of this work was to compare the biomass and community structure of bacteria, protozoa and archaea communities in samples of rumen and faeces of goats and to what extent the diet (alfalfa hay with or without supplemented oats) offered to them exert an influence. Four cannulated adult goats fistulated in the rumen were used in a cross over design experiment in two experimental periods of 26 days, consisting in 14 days of adaptation, 7 days of sampling rumen contents and 5 days of digestibility measurement. Bacterial, protozoa and archaeal biomass and the communities' structure was assessed by real time PCR (qPCR) and denaturing gradient gel electrophoresis (DGGE), respectively. The numbers of archaea and bacteria in both rumen and faeces were higher and lower, respectively, in animals fed AH diet (P < 0.005). Contrary, protozoal numbers were not affected by the diet but were lower (P < 0.001) in faeces than in rumen. The analysis of the community structure revealed a consistently different population in structure in rumen and faeces for the three studied microbial groups and that supplementing alfalfa hay with oats led to a decrease in the similarity between sites in the rumen and faeces: similarity indexes for bacteria (57 and 27%), archaea (26 and 9%) and protozoa (62 and 22%) in animals fed AH and AHO diets, respectively.

  20. Molecular comparative assessment of the microbial ecosystem in rumen and faeces of goats fed alfalfa hay alone or combined with oats.

    PubMed

    Mohammadzadeh, Hamid; Yáñez-Ruiz, David R; Martínez-Fernandez, Gonzalo; Abecia, Leticia

    2014-10-01

    The objective of this work was to compare the biomass and community structure of bacteria, protozoa and archaea communities in samples of rumen and faeces of goats and to what extent the diet (alfalfa hay with or without supplemented oats) offered to them exert an influence. Four cannulated adult goats fistulated in the rumen were used in a cross over design experiment in two experimental periods of 26 days, consisting in 14 days of adaptation, 7 days of sampling rumen contents and 5 days of digestibility measurement. Bacterial, protozoa and archaeal biomass and the communities' structure was assessed by real time PCR (qPCR) and denaturing gradient gel electrophoresis (DGGE), respectively. The numbers of archaea and bacteria in both rumen and faeces were higher and lower, respectively, in animals fed AH diet (P < 0.005). Contrary, protozoal numbers were not affected by the diet but were lower (P < 0.001) in faeces than in rumen. The analysis of the community structure revealed a consistently different population in structure in rumen and faeces for the three studied microbial groups and that supplementing alfalfa hay with oats led to a decrease in the similarity between sites in the rumen and faeces: similarity indexes for bacteria (57 and 27%), archaea (26 and 9%) and protozoa (62 and 22%) in animals fed AH and AHO diets, respectively. PMID:24333680

  1. Mining of luxS genes from rumen microbial consortia by metagenomic and metatranscriptomic approaches.

    PubMed

    Ghali, Ines; Shinkai, Takumi; Mitsumori, Makoto

    2016-05-01

    Although rumen bacterial communities vary depending on many factors such as diet, age and physiological conditions, a core microbiota exists within the rumen. In many natural environments, some bacteria use a quorum-sensing (QS) system to regulate their physiological activities. However, very limited information is available about QS systems in rumen. To investigate the autoinducer 2 (AI-2)-mediated QS system in rumen, we detected genes (luxS) encoding the AI-2 synthase (LuxS), from three datasets embedded in metagenomics RAST server (MG-RAST) and from a metatranscriptome dataset. We collected 135 luxS genes from the metagenomic datasets, which were presumed to originate from Bacteroidetes, Firmicutes, Fusobacteria and Actinobacteria, and 34 luxS genes from the metatranscriptome dataset, which probably originated from Bacteroidetes, Firmicutes and Spirochaetes. Because the essential amino acids for LuxS activity were conserved in the LuxS homologues predicted from luxS gene sequences from both datasets, the LuxS homologues probably function in the rumen. Since the largest number of sequences of luxS genes were collected from the genera Prevotella, Ruminococcus and Eubacterium, which include many fibrolytic bacteria and constituent members of biofilm on feed particles, an AI-2-mediated QS system is likely involved in biofilm formation and fibrolytic activity in the rumen. PMID:26277986

  2. Distribution and Genetic Diversity of Bacteriocin Gene Clusters in Rumen Microbial Genomes

    PubMed Central

    Azevedo, Analice C.; Bento, Cláudia B. P.; Ruiz, Jeronimo C.; Queiroz, Marisa V.

    2015-01-01

    Some species of ruminal bacteria are known to produce antimicrobial peptides, but the screening procedures have mostly been based on in vitro assays using standardized methods. Recent sequencing efforts have made available the genome sequences of hundreds of ruminal microorganisms. In this work, we performed genome mining of the complete and partial genome sequences of 224 ruminal bacteria and 5 ruminal archaea to determine the distribution and diversity of bacteriocin gene clusters. A total of 46 bacteriocin gene clusters were identified in 33 strains of ruminal bacteria. Twenty gene clusters were related to lanthipeptide biosynthesis, while 11 gene clusters were associated with sactipeptide production, 7 gene clusters were associated with class II bacteriocin production, and 8 gene clusters were associated with class III bacteriocin production. The frequency of strains whose genomes encode putative antimicrobial peptide precursors was 14.4%. Clusters related to the production of sactipeptides were identified for the first time among ruminal bacteria. BLAST analysis indicated that the majority of the gene clusters (88%) encoding putative lanthipeptides contained all the essential genes required for lanthipeptide biosynthesis. Most strains of Streptococcus (66.6%) harbored complete lanthipeptide gene clusters, in addition to an open reading frame encoding a putative class II bacteriocin. Albusin B-like proteins were found in 100% of the Ruminococcus albus strains screened in this study. The in silico analysis provided evidence of novel biosynthetic gene clusters in bacterial species not previously related to bacteriocin production, suggesting that the rumen microbiota represents an underexplored source of antimicrobial peptides. PMID:26253660

  3. Distribution and Genetic Diversity of Bacteriocin Gene Clusters in Rumen Microbial Genomes.

    PubMed

    Azevedo, Analice C; Bento, Cláudia B P; Ruiz, Jeronimo C; Queiroz, Marisa V; Mantovani, Hilário C

    2015-10-01

    Some species of ruminal bacteria are known to produce antimicrobial peptides, but the screening procedures have mostly been based on in vitro assays using standardized methods. Recent sequencing efforts have made available the genome sequences of hundreds of ruminal microorganisms. In this work, we performed genome mining of the complete and partial genome sequences of 224 ruminal bacteria and 5 ruminal archaea to determine the distribution and diversity of bacteriocin gene clusters. A total of 46 bacteriocin gene clusters were identified in 33 strains of ruminal bacteria. Twenty gene clusters were related to lanthipeptide biosynthesis, while 11 gene clusters were associated with sactipeptide production, 7 gene clusters were associated with class II bacteriocin production, and 8 gene clusters were associated with class III bacteriocin production. The frequency of strains whose genomes encode putative antimicrobial peptide precursors was 14.4%. Clusters related to the production of sactipeptides were identified for the first time among ruminal bacteria. BLAST analysis indicated that the majority of the gene clusters (88%) encoding putative lanthipeptides contained all the essential genes required for lanthipeptide biosynthesis. Most strains of Streptococcus (66.6%) harbored complete lanthipeptide gene clusters, in addition to an open reading frame encoding a putative class II bacteriocin. Albusin B-like proteins were found in 100% of the Ruminococcus albus strains screened in this study. The in silico analysis provided evidence of novel biosynthetic gene clusters in bacterial species not previously related to bacteriocin production, suggesting that the rumen microbiota represents an underexplored source of antimicrobial peptides.

  4. Distribution and Genetic Diversity of Bacteriocin Gene Clusters in Rumen Microbial Genomes.

    PubMed

    Azevedo, Analice C; Bento, Cláudia B P; Ruiz, Jeronimo C; Queiroz, Marisa V; Mantovani, Hilário C

    2015-10-01

    Some species of ruminal bacteria are known to produce antimicrobial peptides, but the screening procedures have mostly been based on in vitro assays using standardized methods. Recent sequencing efforts have made available the genome sequences of hundreds of ruminal microorganisms. In this work, we performed genome mining of the complete and partial genome sequences of 224 ruminal bacteria and 5 ruminal archaea to determine the distribution and diversity of bacteriocin gene clusters. A total of 46 bacteriocin gene clusters were identified in 33 strains of ruminal bacteria. Twenty gene clusters were related to lanthipeptide biosynthesis, while 11 gene clusters were associated with sactipeptide production, 7 gene clusters were associated with class II bacteriocin production, and 8 gene clusters were associated with class III bacteriocin production. The frequency of strains whose genomes encode putative antimicrobial peptide precursors was 14.4%. Clusters related to the production of sactipeptides were identified for the first time among ruminal bacteria. BLAST analysis indicated that the majority of the gene clusters (88%) encoding putative lanthipeptides contained all the essential genes required for lanthipeptide biosynthesis. Most strains of Streptococcus (66.6%) harbored complete lanthipeptide gene clusters, in addition to an open reading frame encoding a putative class II bacteriocin. Albusin B-like proteins were found in 100% of the Ruminococcus albus strains screened in this study. The in silico analysis provided evidence of novel biosynthetic gene clusters in bacterial species not previously related to bacteriocin production, suggesting that the rumen microbiota represents an underexplored source of antimicrobial peptides. PMID:26253660

  5. Thermodynamic Driving Force of Hydrogen on Rumen Microbial Metabolism: A Theoretical Investigation

    PubMed Central

    van Lingen, Henk J.; Plugge, Caroline M.; Fadel, James G.; Kebreab, Ermias; Bannink, André; Dijkstra, Jan

    2016-01-01

    Hydrogen is a key product of rumen fermentation and has been suggested to thermodynamically control the production of the various volatile fatty acids (VFA). Previous studies, however, have not accounted for the fact that only thermodynamic near-equilibrium conditions control the magnitude of reaction rate. Furthermore, the role of NAD, which is affected by hydrogen partial pressure (PH2), has often not been considered. The aim of this study was to quantify the control of PH2 on reaction rates of specific fermentation pathways, methanogenesis and NADH oxidation in rumen microbes. The control of PH2 was quantified using the thermodynamic potential factor (FT), which is a dimensionless factor that corrects a predicted kinetic reaction rate for the thermodynamic control exerted. Unity FT was calculated for all glucose fermentation pathways considered, indicating no inhibition of PH2 on the production of a specific type of VFA (e.g., acetate, propionate and butyrate) in the rumen. For NADH oxidation without ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity to zero for different NAD+ to NADH ratios and pH of 6.2 and 7.0, which indicates thermodynamic control of PH2. For NADH oxidation with ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity at pH of 7.0 only. For the acetate to propionate conversion, FT increased from 0.65 to unity with increasing PH2, which indicates thermodynamic control. For propionate to acetate and butyrate to acetate conversions, FT decreased to zero below the rumen range of PH2, indicating full thermodynamic suppression. For methanogenesis by archaea without cytochromes, FT differed from unity only below the rumen range of PH2, indicating no thermodynamic control. This theoretical investigation shows that thermodynamic control of PH2 on individual VFA produced and associated yield of hydrogen and methane cannot be explained without considering NADH

  6. Changes in Rumen Microbial Community Composition during Adaption to an In Vitro System and the Impact of Different Forages.

    PubMed

    Lengowski, Melanie B; Zuber, Karin H R; Witzig, Maren; Möhring, Jens; Boguhn, Jeannette; Rodehutscord, Markus

    2016-01-01

    This study examined ruminal microbial community composition alterations during initial adaption to and following incubation in a rumen simulation system (Rusitec) using grass or corn silage as substrates. Samples were collected from fermenter liquids at 0, 2, 4, 12, 24, and 48 h and from feed residues at 0, 24, and 48 h after initiation of incubation (period 1) and on day 13 (period 2). Microbial DNA was extracted and real-time qPCR was used to quantify differences in the abundance of protozoa, methanogens, total bacteria, Fibrobacter succinogenes, Ruminococcus albus, Ruminobacter amylophilus, Prevotella bryantii, Selenomonas ruminantium, and Clostridium aminophilum. We found that forage source and sampling time significantly influenced the ruminal microbial community. The gene copy numbers of most microbial species (except C. aminophilum) decreased in period 1; however, adaption continued through period 2 for several species. The addition of fresh substrate in period 2 led to increasing copy numbers of all microbial species during the first 2-4 h in the fermenter liquid except protozoa, which showed a postprandial decrease. Corn silage enhanced the growth of R. amylophilus and F. succinogenes, and grass silage enhanced R. albus, P. bryantii, and C. aminophilum. No effect of forage source was detected on total bacteria, protozoa, S. ruminantium, or methanogens or on total gas production, although grass silage enhanced methane production. This study showed that the Rusitec provides a stable system after an adaption phase that should last longer than 48 h, and that the forage source influenced several microbial species. PMID:26928330

  7. Changes in Rumen Microbial Community Composition during Adaption to an In Vitro System and the Impact of Different Forages

    PubMed Central

    Lengowski, Melanie B.; Zuber, Karin H. R.; Witzig, Maren; Möhring, Jens; Boguhn, Jeannette; Rodehutscord, Markus

    2016-01-01

    This study examined ruminal microbial community composition alterations during initial adaption to and following incubation in a rumen simulation system (Rusitec) using grass or corn silage as substrates. Samples were collected from fermenter liquids at 0, 2, 4, 12, 24, and 48 h and from feed residues at 0, 24, and 48 h after initiation of incubation (period 1) and on day 13 (period 2). Microbial DNA was extracted and real-time qPCR was used to quantify differences in the abundance of protozoa, methanogens, total bacteria, Fibrobacter succinogenes, Ruminococcus albus, Ruminobacter amylophilus, Prevotella bryantii, Selenomonas ruminantium, and Clostridium aminophilum. We found that forage source and sampling time significantly influenced the ruminal microbial community. The gene copy numbers of most microbial species (except C. aminophilum) decreased in period 1; however, adaption continued through period 2 for several species. The addition of fresh substrate in period 2 led to increasing copy numbers of all microbial species during the first 2–4 h in the fermenter liquid except protozoa, which showed a postprandial decrease. Corn silage enhanced the growth of R. amylophilus and F. succinogenes, and grass silage enhanced R. albus, P. bryantii, and C. aminophilum. No effect of forage source was detected on total bacteria, protozoa, S. ruminantium, or methanogens or on total gas production, although grass silage enhanced methane production. This study showed that the Rusitec provides a stable system after an adaption phase that should last longer than 48 h, and that the forage source influenced several microbial species. PMID:26928330

  8. Microbial Protein-tyrosine Kinases*

    PubMed Central

    Chao, Joseph D.; Wong, Dennis; Av-Gay, Yossef

    2014-01-01

    Microbial ester kinases identified in the past 3 decades came as a surprise, as protein phosphorylation on Ser, Thr, and Tyr amino acids was thought to be unique to eukaryotes. Current analysis of available microbial genomes reveals that “eukaryote-like” protein kinases are prevalent in prokaryotes and can converge in the same signaling pathway with the classical microbial “two-component” systems. Most microbial tyrosine kinases lack the “eukaryotic” Hanks domain signature and are designated tyrosine kinases based upon their biochemical activity. These include the tyrosine kinases termed bacterial tyrosine kinases (BY-kinases), which are responsible for the majority of known bacterial tyrosine phosphorylation events. Although termed generally as bacterial tyrosine kinases, BY-kinases can be considered as one family belonging to the superfamily of prokaryotic protein-tyrosine kinases in bacteria. Other members of this superfamily include atypical “odd” tyrosine kinases with diverse mechanisms of protein phosphorylation and the “eukaryote-like” Hanks-type tyrosine kinases. Here, we discuss the distribution, phylogeny, and function of the various prokaryotic protein-tyrosine kinases, focusing on the recently discovered Mycobacterium tuberculosis PtkA and its relationship with other members of this diverse family of proteins. PMID:24554699

  9. Effects of Phytoecdysteroids (PEDS) Extracted from Cyanotis arachnoidea on Rumen Fermentation, Enzyme Activity and Microbial Efficiency in a Continuous-Culture System

    PubMed Central

    Li, Deyong; Zhang, Yawei; Cui, Zhenliang; He, Liwen; Chen, Wanbao; Meng, Qingxiang; Ren, Liping

    2016-01-01

    The objective of this study was to evaluate the effects of supplementation of phytoecdysteroids (PEDS) extracted from Cyanotis arachnoidea on rumen fermentation, enzymes activity and microbial efficiency in a dual flow continuous-culture system. A single-factor experimental design was used with twelve fermenters in 4 groups with 3 replicates each. Fermenters were incubated for a total of 7 days that included first 4 days for adaptation and last 3 days for sampling. PEDS was added at levels of zero (as control), 5, 10, and 15 mg/g of the substrate (DM). The results showed that increasing supplementation levels of PEDS resulted in incremental digestibility of dry matter (DMD) (quadratic, P = 0.001) and organic matter (OMD) (quadratic, P = 0.031), but unchanged digestibility of neutral detergent fiber (NDFD), crude protein (CPD) and acid detergent acid (ADFD). As supplementation levels of PEDS increased, there were decreased response in the concentration of ammonia nitrogen (NH3-N) (linear, P = 0.015) and increased response in molar proportions of butyrate (linear, P = 0.004), but unchanged response in total volatile fatty acid (TVFA) and the molar proportion of acetate and propionate, respectively. Increasing PEDS supplementation levels decreased the ratio of acetate to propionate (linear, P = 0.038), suggesting an alteration of rumen fermentation pattern occurring due to PEDS supplementation in the diet. Supplementation of PEDS significantly increased activities of glutamate dehydrogenase (quadratic, P = 0.001), alanine dehydrogenase (quadratic, P = 0.004), glutamate synthase (linear, P = 0.038), glutamine synthetase (quadratic, P = 0.011), respectively. There were no discernible differences in the activity of carboxymethyl cellulose (CMCase), xylanase and protease regardless of the treatments. The daily production of microbial nitrogen (linear, P = 0.002) and microbial efficiency (MOEEF) (linear, P = 0.001) increased linearly as supplementation levels of PEDS

  10. Effects of Phytoecdysteroids (PEDS) Extracted from Cyanotis arachnoidea on Rumen Fermentation, Enzyme Activity and Microbial Efficiency in a Continuous-Culture System.

    PubMed

    Li, Deyong; Zhang, Yawei; Cui, Zhenliang; He, Liwen; Chen, Wanbao; Meng, Qingxiang; Ren, Liping

    2016-01-01

    The objective of this study was to evaluate the effects of supplementation of phytoecdysteroids (PEDS) extracted from Cyanotis arachnoidea on rumen fermentation, enzymes activity and microbial efficiency in a dual flow continuous-culture system. A single-factor experimental design was used with twelve fermenters in 4 groups with 3 replicates each. Fermenters were incubated for a total of 7 days that included first 4 days for adaptation and last 3 days for sampling. PEDS was added at levels of zero (as control), 5, 10, and 15 mg/g of the substrate (DM). The results showed that increasing supplementation levels of PEDS resulted in incremental digestibility of dry matter (DMD) (quadratic, P = 0.001) and organic matter (OMD) (quadratic, P = 0.031), but unchanged digestibility of neutral detergent fiber (NDFD), crude protein (CPD) and acid detergent acid (ADFD). As supplementation levels of PEDS increased, there were decreased response in the concentration of ammonia nitrogen (NH3-N) (linear, P = 0.015) and increased response in molar proportions of butyrate (linear, P = 0.004), but unchanged response in total volatile fatty acid (TVFA) and the molar proportion of acetate and propionate, respectively. Increasing PEDS supplementation levels decreased the ratio of acetate to propionate (linear, P = 0.038), suggesting an alteration of rumen fermentation pattern occurring due to PEDS supplementation in the diet. Supplementation of PEDS significantly increased activities of glutamate dehydrogenase (quadratic, P = 0.001), alanine dehydrogenase (quadratic, P = 0.004), glutamate synthase (linear, P = 0.038), glutamine synthetase (quadratic, P = 0.011), respectively. There were no discernible differences in the activity of carboxymethyl cellulose (CMCase), xylanase and protease regardless of the treatments. The daily production of microbial nitrogen (linear, P = 0.002) and microbial efficiency (MOEEF) (linear, P = 0.001) increased linearly as supplementation levels of PEDS

  11. Effects of Phytoecdysteroids (PEDS) Extracted from Cyanotis arachnoidea on Rumen Fermentation, Enzyme Activity and Microbial Efficiency in a Continuous-Culture System.

    PubMed

    Li, Deyong; Zhang, Yawei; Cui, Zhenliang; He, Liwen; Chen, Wanbao; Meng, Qingxiang; Ren, Liping

    2016-01-01

    The objective of this study was to evaluate the effects of supplementation of phytoecdysteroids (PEDS) extracted from Cyanotis arachnoidea on rumen fermentation, enzymes activity and microbial efficiency in a dual flow continuous-culture system. A single-factor experimental design was used with twelve fermenters in 4 groups with 3 replicates each. Fermenters were incubated for a total of 7 days that included first 4 days for adaptation and last 3 days for sampling. PEDS was added at levels of zero (as control), 5, 10, and 15 mg/g of the substrate (DM). The results showed that increasing supplementation levels of PEDS resulted in incremental digestibility of dry matter (DMD) (quadratic, P = 0.001) and organic matter (OMD) (quadratic, P = 0.031), but unchanged digestibility of neutral detergent fiber (NDFD), crude protein (CPD) and acid detergent acid (ADFD). As supplementation levels of PEDS increased, there were decreased response in the concentration of ammonia nitrogen (NH3-N) (linear, P = 0.015) and increased response in molar proportions of butyrate (linear, P = 0.004), but unchanged response in total volatile fatty acid (TVFA) and the molar proportion of acetate and propionate, respectively. Increasing PEDS supplementation levels decreased the ratio of acetate to propionate (linear, P = 0.038), suggesting an alteration of rumen fermentation pattern occurring due to PEDS supplementation in the diet. Supplementation of PEDS significantly increased activities of glutamate dehydrogenase (quadratic, P = 0.001), alanine dehydrogenase (quadratic, P = 0.004), glutamate synthase (linear, P = 0.038), glutamine synthetase (quadratic, P = 0.011), respectively. There were no discernible differences in the activity of carboxymethyl cellulose (CMCase), xylanase and protease regardless of the treatments. The daily production of microbial nitrogen (linear, P = 0.002) and microbial efficiency (MOEEF) (linear, P = 0.001) increased linearly as supplementation levels of PEDS

  12. A novel feruloyl esterase from rumen microbial metagenome: Gene cloning and enzyme characterization in the release of mono- and diferulic acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A feruloyl esterase (FAE) gene was isolated from a rumen microbial metagenome, cloned into E. coli, and expressed in active form. The enzyme (RuFae4) was classified as a Type D feruloyl esterase based on its action on synthetic substrates and ability to release diferulates. The RuFae4 alone releas...

  13. Rumen fermentation and microbial population in lactating dairy cows receiving diets containing oilseeds rich in C-18 fatty acids.

    PubMed

    Ivan, M; Petit, H V; Chiquette, J; Wright, A-D G

    2013-04-14

    Sixteen Holstein rumen-cannulated primiparous milking dairy cows were fed a control diet (CN) based on maize silage and soyabean meal during a 4-week period before the start of a 21-d experiment with oilseeds containing high concentration of linoleic acid (Linola™) or linolenic acid (NuLin™). Thereafter, four cows received ad libitum one of each of four dietary treatments comprising of CN, Linola (LN), NuLin (NL) and LN/NL (50/50 % combination). Each LN, NL and LN/NL treatment contained 6 % oil of DM. Rumen digesta samples were collected on days 6, 11, 16 and 21 and milk samples on days 13, 15 and 17. There were no effects (P>0.05) of the oilseeds on pH and concentrations of NH3-N and total volatile fatty acids, while the acetate:propionate ratio was decreased (P< 0.05). The oilseeds also decreased (P< 0.05) protozoa and increased (P< 0.1) total cellulolytic bacteria in rumen fluid, especially when containing high dietary linoleic acid (P< 0.05). The milk protein concentration was increased (P< 0.1) by the dietary linoleic acid, which produced most beneficial results. It was concluded that supplements of linoleic acid in diets of ruminants might contribute to better digestion of dietary fibre and increased quality of milk.

  14. A comparison of rumen microbial profiles in dairy cows as retrieved by 454 Roche and Ion Torrent (PGM) sequencing platforms

    PubMed Central

    Indugu, Nagaraju; Bittinger, Kyle; Kumar, Sanjay; Vecchiarelli, Bonnie

    2016-01-01

    Next generation sequencing (NGS) technology is a widely accepted tool used by microbial ecologists to explore complex microbial communities in different ecosystems. As new NGS platforms continue to become available, it becomes imperative to compare data obtained from different platforms and analyze their effect on microbial community structure. In the present study, we compared sequencing data from both the 454 and Ion Torrent (PGM) platforms on the same DNA samples obtained from the rumen of dairy cows during their transition period. Despite the substantial difference in the number of reads, error rate and length of reads among both platforms, we identified similar community composition between the two data sets. Procrustes analysis revealed similar correlations (M2 = 0.319; P = 0.001) in the microbial community composition between the two platforms. Both platforms revealed the abundance of the same bacterial phyla which were Bacteroidetes and Firmicutes; however, PGM recovered an additional four phyla. Comparisons made at the genus level by each platforms revealed differences in only a few genera such as Prevotella, Ruminococcus, Succiniclasticum and Treponema (p < 0.05; chi square test). Collectively, we conclude that the output generated from PGM and 454 yielded concurrent results, provided stringent bioinformatics pipelines are employed. PMID:26870608

  15. A comparison of rumen microbial profiles in dairy cows as retrieved by 454 Roche and Ion Torrent (PGM) sequencing platforms.

    PubMed

    Indugu, Nagaraju; Bittinger, Kyle; Kumar, Sanjay; Vecchiarelli, Bonnie; Pitta, Dipti

    2016-01-01

    Next generation sequencing (NGS) technology is a widely accepted tool used by microbial ecologists to explore complex microbial communities in different ecosystems. As new NGS platforms continue to become available, it becomes imperative to compare data obtained from different platforms and analyze their effect on microbial community structure. In the present study, we compared sequencing data from both the 454 and Ion Torrent (PGM) platforms on the same DNA samples obtained from the rumen of dairy cows during their transition period. Despite the substantial difference in the number of reads, error rate and length of reads among both platforms, we identified similar community composition between the two data sets. Procrustes analysis revealed similar correlations (M (2) = 0.319; P = 0.001) in the microbial community composition between the two platforms. Both platforms revealed the abundance of the same bacterial phyla which were Bacteroidetes and Firmicutes; however, PGM recovered an additional four phyla. Comparisons made at the genus level by each platforms revealed differences in only a few genera such as Prevotella, Ruminococcus, Succiniclasticum and Treponema (p < 0.05; chi square test). Collectively, we conclude that the output generated from PGM and 454 yielded concurrent results, provided stringent bioinformatics pipelines are employed. PMID:26870608

  16. Effects of rare earth element lanthanum on rumen methane and volatile fatty acid production and microbial flora in vitro.

    PubMed

    Zhang, T T; Zhao, G Y; Zheng, W S; Niu, W J; Wei, C; Lin, S X

    2015-06-01

    The objectives of the trial were to study the effects of rare earth element (REE) lanthanum (La) on the in vitro rumen methane (CH4 ) and volatile fatty acid (VFA) production and the microbial flora of feeds. Four feed mixtures with different levels of neutral detergent fibre (NDF), that is 20.0% (I), 31.0% (II), 41.9% (III) and 52.7% (IV), were formulated as substrates. Five levels of LaCl3 , that is 0, 0.4, 0.6, 0.8 and 1.0 mmol/kg dry matter (DM), were added to the feed mixtures, respectively, as experimental treatments in a two-factor 5 × 4 randomized design. The in vitro incubation lasted for 24 h. The results showed that supplementing LaCl3 increased the total gas (p < 0.001) production and tended to increase the total VFA production (p = 0.072) and decreased the CH4 production (p = 0.001) and the ratios of acetate/propionate (p = 0.019) and CH4 /total VFA (p < 0.001). Interactions between LaCl3 and NDF were significant in total gas production (p = 0.030) and tended to be significant in CH4 production (p = 0.071). Supplementing LaCl3 at the level of 0.8 mmol/g DM decreased the relative abundance of methanogens and protozoa in the total bacterial 16S rDNA analysed using the real-time PCR (p < 0.0001), increased F. succinogenes (p = 0.0003) and decreased R. flavefaciens (p < 0.0001) whereas did not affect R. albus and anaerobic fungi (p > 0.05). It was concluded that LaCl3 decreased the CH4 production without negatively affecting feed digestion through manipulating rumen microbial flora when feed mixtures with different levels of NDF were used as substrates.

  17. Characterization of the rumen microbiota of pre-ruminant calves using metagenomic tools.

    PubMed

    Li, Robert W; Connor, Erin E; Li, Congjun; Baldwin Vi, Ransom L; Sparks, Michael E

    2012-01-01

    The temporal sequence of microbial establishment in the rumen of the neonatal ruminant has important ecological and pathophysiological implications. In this study, we characterized the rumen microbiota of pre-ruminant calves fed milk replacer using two approaches, pyrosequencing of hypervariable V3-V5 regions of the 16S rRNA gene and whole-genome shotgun approach. Fifteen bacterial phyla were identified in the microbiota of pre-ruminant calves. Bacteroidetes was the predominant phylum in the rumen microbiota of 42-day-old calves, representing 74.8% of the 16S sequences, followed by Firmicutes (12.0%), Proteobacteria (10.4%), Verrucomicrobia (1.2%) and Synergistetes (1.1%). However, the phylum-level composition of 14-day-old calves was distinctly different. A total of 170 bacterial genera were identified while the core microbiome of pre-ruminant calves included 45 genera. Rumen development seemingly had a significant impact on microbial diversity. The dazzling functional diversity of the rumen microbiota was reflected by identification of 8298 Pfam and 3670 COG protein families. The rumen microbiota of pre-ruminant calves displayed a considerable compositional heterogeneity during early development. This is evidenced by a profound difference in rumen microbial composition between the two age groups. However, all functional classes between the two age groups had a remarkably similar assignment, suggesting that rumen microbial communities of pre-ruminant calves maintained a stable function and metabolic potentials while their phylogenetic composition fluctuated greatly. The presence of all major types of rumen microorganisms suggests that the rumen of pre-ruminant calves may not be rudimentary. Our results provide insight into rumen microbiota dynamics and will facilitate efforts in formulating optimal early-weaning strategies.

  18. Characterization of the rumen microbiota of pre-ruminant calves using metagenomic tools.

    PubMed

    Li, Robert W; Connor, Erin E; Li, Congjun; Baldwin Vi, Ransom L; Sparks, Michael E

    2012-01-01

    The temporal sequence of microbial establishment in the rumen of the neonatal ruminant has important ecological and pathophysiological implications. In this study, we characterized the rumen microbiota of pre-ruminant calves fed milk replacer using two approaches, pyrosequencing of hypervariable V3-V5 regions of the 16S rRNA gene and whole-genome shotgun approach. Fifteen bacterial phyla were identified in the microbiota of pre-ruminant calves. Bacteroidetes was the predominant phylum in the rumen microbiota of 42-day-old calves, representing 74.8% of the 16S sequences, followed by Firmicutes (12.0%), Proteobacteria (10.4%), Verrucomicrobia (1.2%) and Synergistetes (1.1%). However, the phylum-level composition of 14-day-old calves was distinctly different. A total of 170 bacterial genera were identified while the core microbiome of pre-ruminant calves included 45 genera. Rumen development seemingly had a significant impact on microbial diversity. The dazzling functional diversity of the rumen microbiota was reflected by identification of 8298 Pfam and 3670 COG protein families. The rumen microbiota of pre-ruminant calves displayed a considerable compositional heterogeneity during early development. This is evidenced by a profound difference in rumen microbial composition between the two age groups. However, all functional classes between the two age groups had a remarkably similar assignment, suggesting that rumen microbial communities of pre-ruminant calves maintained a stable function and metabolic potentials while their phylogenetic composition fluctuated greatly. The presence of all major types of rumen microorganisms suggests that the rumen of pre-ruminant calves may not be rudimentary. Our results provide insight into rumen microbiota dynamics and will facilitate efforts in formulating optimal early-weaning strategies. PMID:21906219

  19. Bovine Host Genetic Variation Influences Rumen Microbial Methane Production with Best Selection Criterion for Low Methane Emitting and Efficiently Feed Converting Hosts Based on Metagenomic Gene Abundance.

    PubMed

    Roehe, Rainer; Dewhurst, Richard J; Duthie, Carol-Anne; Rooke, John A; McKain, Nest; Ross, Dave W; Hyslop, Jimmy J; Waterhouse, Anthony; Freeman, Tom C; Watson, Mick; Wallace, R John

    2016-02-01

    Methane produced by methanogenic archaea in ruminants contributes significantly to anthropogenic greenhouse gas emissions. The host genetic link controlling microbial methane production is unknown and appropriate genetic selection strategies are not developed. We used sire progeny group differences to estimate the host genetic influence on rumen microbial methane production in a factorial experiment consisting of crossbred breed types and diets. Rumen metagenomic profiling was undertaken to investigate links between microbial genes and methane emissions or feed conversion efficiency. Sire progeny groups differed significantly in their methane emissions measured in respiration chambers. Ranking of the sire progeny groups based on methane emissions or relative archaeal abundance was consistent overall and within diet, suggesting that archaeal abundance in ruminal digesta is under host genetic control and can be used to genetically select animals without measuring methane directly. In the metagenomic analysis of rumen contents, we identified 3970 microbial genes of which 20 and 49 genes were significantly associated with methane emissions and feed conversion efficiency respectively. These explained 81% and 86% of the respective variation and were clustered in distinct functional gene networks. Methanogenesis genes (e.g. mcrA and fmdB) were associated with methane emissions, whilst host-microbiome cross talk genes (e.g. TSTA3 and FucI) were associated with feed conversion efficiency. These results strengthen the idea that the host animal controls its own microbiota to a significant extent and open up the implementation of effective breeding strategies using rumen microbial gene abundance as a predictor for difficult-to-measure traits on a large number of hosts. Generally, the results provide a proof of principle to use the relative abundance of microbial genes in the gastrointestinal tract of different species to predict their influence on traits e.g. human metabolism

  20. Bovine Host Genetic Variation Influences Rumen Microbial Methane Production with Best Selection Criterion for Low Methane Emitting and Efficiently Feed Converting Hosts Based on Metagenomic Gene Abundance

    PubMed Central

    Roehe, Rainer; Dewhurst, Richard J.; Duthie, Carol-Anne; Rooke, John A.; McKain, Nest; Ross, Dave W.; Hyslop, Jimmy J.; Waterhouse, Anthony; Freeman, Tom C.

    2016-01-01

    Methane produced by methanogenic archaea in ruminants contributes significantly to anthropogenic greenhouse gas emissions. The host genetic link controlling microbial methane production is unknown and appropriate genetic selection strategies are not developed. We used sire progeny group differences to estimate the host genetic influence on rumen microbial methane production in a factorial experiment consisting of crossbred breed types and diets. Rumen metagenomic profiling was undertaken to investigate links between microbial genes and methane emissions or feed conversion efficiency. Sire progeny groups differed significantly in their methane emissions measured in respiration chambers. Ranking of the sire progeny groups based on methane emissions or relative archaeal abundance was consistent overall and within diet, suggesting that archaeal abundance in ruminal digesta is under host genetic control and can be used to genetically select animals without measuring methane directly. In the metagenomic analysis of rumen contents, we identified 3970 microbial genes of which 20 and 49 genes were significantly associated with methane emissions and feed conversion efficiency respectively. These explained 81% and 86% of the respective variation and were clustered in distinct functional gene networks. Methanogenesis genes (e.g. mcrA and fmdB) were associated with methane emissions, whilst host-microbiome cross talk genes (e.g. TSTA3 and FucI) were associated with feed conversion efficiency. These results strengthen the idea that the host animal controls its own microbiota to a significant extent and open up the implementation of effective breeding strategies using rumen microbial gene abundance as a predictor for difficult-to-measure traits on a large number of hosts. Generally, the results provide a proof of principle to use the relative abundance of microbial genes in the gastrointestinal tract of different species to predict their influence on traits e.g. human metabolism

  1. Bovine Host Genetic Variation Influences Rumen Microbial Methane Production with Best Selection Criterion for Low Methane Emitting and Efficiently Feed Converting Hosts Based on Metagenomic Gene Abundance.

    PubMed

    Roehe, Rainer; Dewhurst, Richard J; Duthie, Carol-Anne; Rooke, John A; McKain, Nest; Ross, Dave W; Hyslop, Jimmy J; Waterhouse, Anthony; Freeman, Tom C; Watson, Mick; Wallace, R John

    2016-02-01

    Methane produced by methanogenic archaea in ruminants contributes significantly to anthropogenic greenhouse gas emissions. The host genetic link controlling microbial methane production is unknown and appropriate genetic selection strategies are not developed. We used sire progeny group differences to estimate the host genetic influence on rumen microbial methane production in a factorial experiment consisting of crossbred breed types and diets. Rumen metagenomic profiling was undertaken to investigate links between microbial genes and methane emissions or feed conversion efficiency. Sire progeny groups differed significantly in their methane emissions measured in respiration chambers. Ranking of the sire progeny groups based on methane emissions or relative archaeal abundance was consistent overall and within diet, suggesting that archaeal abundance in ruminal digesta is under host genetic control and can be used to genetically select animals without measuring methane directly. In the metagenomic analysis of rumen contents, we identified 3970 microbial genes of which 20 and 49 genes were significantly associated with methane emissions and feed conversion efficiency respectively. These explained 81% and 86% of the respective variation and were clustered in distinct functional gene networks. Methanogenesis genes (e.g. mcrA and fmdB) were associated with methane emissions, whilst host-microbiome cross talk genes (e.g. TSTA3 and FucI) were associated with feed conversion efficiency. These results strengthen the idea that the host animal controls its own microbiota to a significant extent and open up the implementation of effective breeding strategies using rumen microbial gene abundance as a predictor for difficult-to-measure traits on a large number of hosts. Generally, the results provide a proof of principle to use the relative abundance of microbial genes in the gastrointestinal tract of different species to predict their influence on traits e.g. human metabolism

  2. Rumen Degradability and Small Intestinal Digestibility of the Amino Acids in Four Protein Supplements

    PubMed Central

    Wang, Y.; Jin, L.; Wen, Q. N.; Kopparapu, N. K.; Liu, J.; Liu, X. L.; Zhang, Y. G.

    2016-01-01

    The supplementation of livestock feed with animal protein is a present cause for public concern, and plant protein shortages have become increasingly prominent in China. This conflict may be resolved by fully utilizing currently available sources of plant protein. We estimated the rumen degradability and the small intestinal digestibility of the amino acids (AA) in rapeseed meal (RSM), soybean meal (SBM), sunflower seed meal (SFM) and sesame meal (SSM) using the mobile nylon bag method to determine the absorbable AA content of these protein supplements as a guide towards dietary formulations for the dairy industry. Overall, this study aimed to utilize protein supplements effectively to guide dietary formulations to increase milk yield and save plant protein resources. To this end, we studied four cows with a permanent rumen fistula and duodenal T-shape fistula in a 4×4 Latin square experimental design. The results showed that the total small intestine absorbable amino acids and small intestine absorbable essential amino acids were higher in the SBM (26.34% and 13.11% dry matter [DM], respectively) than in the SFM (13.97% and 6.89% DM, respectively). The small intestine absorbable Lys contents of the SFM, SSM, RSM and SBM were 0.86%, 0.88%, 1.43%, and 2.12% (DM basis), respectively, and the absorbable Met contents of these meals were 0.28%, 1.03%, 0.52%, and 0.47% (DM basis), respectively. Among the examined food sources, the milk protein score of the SBM (0.181) was highest followed by those of the RSM (0.136), SSM (0.108) and SFM (0.106). The absorbable amino acid contents of the protein supplements accurately reflected protein availability, which is an important indicator of the balance of feed formulation. Therefore, a database detailing the absorbable AA should be established. PMID:26732449

  3. Rumen Degradability and Small Intestinal Digestibility of the Amino Acids in Four Protein Supplements.

    PubMed

    Wang, Y; Jin, L; Wen, Q N; Kopparapu, N K; Liu, J; Liu, X L; Zhang, Y G

    2016-02-01

    The supplementation of livestock feed with animal protein is a present cause for public concern, and plant protein shortages have become increasingly prominent in China. This conflict may be resolved by fully utilizing currently available sources of plant protein. We estimated the rumen degradability and the small intestinal digestibility of the amino acids (AA) in rapeseed meal (RSM), soybean meal (SBM), sunflower seed meal (SFM) and sesame meal (SSM) using the mobile nylon bag method to determine the absorbable AA content of these protein supplements as a guide towards dietary formulations for the dairy industry. Overall, this study aimed to utilize protein supplements effectively to guide dietary formulations to increase milk yield and save plant protein resources. To this end, we studied four cows with a permanent rumen fistula and duodenal T-shape fistula in a 4×4 Latin square experimental design. The results showed that the total small intestine absorbable amino acids and small intestine absorbable essential amino acids were higher in the SBM (26.34% and 13.11% dry matter [DM], respectively) than in the SFM (13.97% and 6.89% DM, respectively). The small intestine absorbable Lys contents of the SFM, SSM, RSM and SBM were 0.86%, 0.88%, 1.43%, and 2.12% (DM basis), respectively, and the absorbable Met contents of these meals were 0.28%, 1.03%, 0.52%, and 0.47% (DM basis), respectively. Among the examined food sources, the milk protein score of the SBM (0.181) was highest followed by those of the RSM (0.136), SSM (0.108) and SFM (0.106). The absorbable amino acid contents of the protein supplements accurately reflected protein availability, which is an important indicator of the balance of feed formulation. Therefore, a database detailing the absorbable AA should be established.

  4. Vaccination of cattle with a methanogen protein produces specific antibodies in the saliva which are stable in the rumen.

    PubMed

    Subharat, Supatsak; Shu, Dairu; Zheng, Tao; Buddle, Bryce M; Janssen, Peter H; Luo, Dongwen; Wedlock, D Neil

    2015-04-15

    Methane is produced in the rumen of cattle by a group of archaea (single-celled organisms forming a domain distinct from bacteria and eucarya) called methanogens. Vaccination against methanogens has the potential to reduce methane emissions by inducing antibodies in saliva which are transferred to the rumen and diminish the ability of methanogens to produce methane. Since it is likely that an effective vaccination strategy will need to produce high levels of methanogen-specific antibody in the saliva; the choice of adjuvant, route of vaccination and stability of saliva-derived antibody in the rumen all need to be considered. In this study, stability of IgA and IgG in rumen fluid was determined using an in vitro assay. IgA levels in cattle saliva were reduced by only 40% after 8h exposure to rumen contents while IgG levels were reduced by 80%. These results indicated that antibody is relatively stable in the bovine rumen. A trial was conducted in cattle to investigate induction of immune responses to a methanogen protein, recombinant glycosyl transferase protein (rGT2) from Methanobrevibacter ruminantium M1. Groups of cattle (n=6) were vaccinated subcutaneously with rGT2, formulated with Montanide ISA61 with or without the TLR4 agonist, monophosphoryl lipid A (MPL). A control group (n=6) was not vaccinated. Strong antigen-specific IgG and moderate IgA responses were measured in the serum and saliva of the vaccinated animals and antibody was also detected in the rumen.

  5. Milk fatty acid composition, rumen microbial population, and animal performances in response to diets rich in linoleic acid supplemented with chestnut or quebracho tannins in dairy ewes.

    PubMed

    Buccioni, A; Pauselli, M; Viti, C; Minieri, S; Pallara, G; Roscini, V; Rapaccini, S; Marinucci, M Trabalza; Lupi, P; Conte, G; Mele, M

    2015-02-01

    The aim of the study was to evaluate milk fatty acid (FA) profile, animal performance, and rumen microbial population in response to diets containing soybean oil supplemented or not with chestnut and quebracho tannins in dairy ewes. Eighteen Comisana ewes at 122±6 d in milking were allotted into 3 experimental groups. Diets were characterized by chopped grass hay administered ad libitum and by 800 g/head and day of 3 experimental concentrates containing 84.5 g of soybean oil/kg of dry matter (DM) and 52.8 g/kg of DM of bentonite (control diet), chestnut tannin extract (CHT diet), or quebracho tannin extract (QUE diet). The trial lasted 4 wk. Milk yield was recorded daily, and milk composition and blood parameters were analyzed weekly. At the end of the experiment, samples of rumen fluid were collected to analyze pH, volatile fatty acid profile, and the relative proportions of Butyrivibrio fibrisolvens and Butyrivibrio proteoclasticus in the rumen microbial population. Hepatic functionality, milk yield, and gross composition were not affected by tannin extracts, whereas milk FA composition was characterized by significant changes in the concentration of linoleic acid (CHT +2.77% and QUE +9.23%), vaccenic acid (CHT +7.07% and QUE +13.88%), rumenic acid (CHT -1.88% and QUE +24.24%), stearic acid (CHT + 8.71% and QUE -11.45%), and saturated fatty acids (CHT -0.47% and QUE -3.38%). These differences were probably due to the ability of condensed versus hydrolyzable tannins to interfere with rumen microbial metabolism, as indirectly confirmed by changes in the relative proportions of B. fibrisolvens and B. proteoclasticus populations and by changes in the molar proportions of volatile fatty acids. The effect of the CHT diet on the milk FA profile and microbial species considered in this trial was intermediate between that of QUE and the control diet, suggesting a differential effect of condensed and hydrolyzable tannins on rumen microbes. Compared with control animals

  6. Effect of protein supplementation on ruminal parameters and microbial community fingerprint of Nellore steers fed tropical forages.

    PubMed

    Bento, C B P; Azevedo, A C; Gomes, D I; Batista, E D; Rufino, L M A; Detmann, E; Mantovani, H C

    2016-01-01

    In tropical regions, protein supplementation is a common practice in dairy and beef farming. However, the effect of highly degradable protein in ruminal fermentation and microbial community composition has not yet been investigated in a systematic manner. In this work, we aimed to investigate the impact of casein supplementation on volatile fatty acids (VFA) production, specific activity of deamination (SAD), ammonia concentration and bacterial and archaeal community composition. The experimental design was a 4×4 Latin square balanced for residual effects, with four animals (average initial weight of 280±10 kg) and four experimental periods, each with duration of 29 days. The diet comprised Tifton 85 (Cynodon sp.) hay with an average CP content of 9.8%, on a dry matter basis. Animals received basal forage (control) or infusions of pure casein (230 g) administered direct into the rumen, abomasum or divided (50 : 50 ratio) in the rumen/abomasum. There was no differences (P>0.05) in ruminal pH and microbial protein concentration between supplemented v. non-supplemented animals. However, in steers receiving ruminal infusion of casein the SAD and ruminal ammonia concentration increased 33% and 76%, respectively, compared with the control. The total concentration of VFA increased (P0.05) in species richness and diversity of γ-proteobacteria, firmicutes and archaea between non-supplemented Nellore steers and steers receiving casein supplementation in the rumen. However, species richness and the Shannon-Wiener index were lower (P<0.05) for the phylum bacteroidetes in steers supplemented with casein in the rumen compared with non-supplemented animals. Venn diagrams indicated that the number of unique bands varied considerably among individual animals and was usually higher in number for non-supplemented steers compared with supplemented animals. These results add new knowledge about the effects of ruminal and postruminal protein supplementation on metabolic activities of

  7. From rumen to industry.

    PubMed

    Sauer, Michael; Marx, Hans; Mattanovich, Diethard

    2012-09-10

    The rumen is one of the most complicated and most fascinating microbial ecosystems in nature. A wide variety of microbial species, including bacteria, fungi and protozoa act together to bioconvert (ligno)cellulosic plant material into compounds, which can be taken up and metabolized by the ruminant. Thus, the rumen perfectly resembles a solution to a current industrial problem: the biorefinery, which aims at the bioconversion of lignocellulosic material into fuels and chemicals. We suggest to intensify the studies of the ruminal microbial ecosystem from an industrial microbiologists point of view in order to make use of this rich source of organisms and enzymes.

  8. Effect of Saccharomyces cerevisiae on alfalfa nutrient degradation characteristics and rumen microbial populations of steers fed diets with different concentrate-to-forage ratios

    PubMed Central

    2014-01-01

    Live yeast (Saccharomyces cerevisiae) constitutes an effective additive for animal production; its probiotic effect may be related to the concentrate-to-forage ratio (CTFR). The objective of this study was to assess the effects of S. cerevisiae (SC) on fiber degradation and rumen microbial populations in steers fed diets with different levels of dietary concentrate. Ten Simmental × Local crossbred steers (450 ± 50 kg BW) were assigned to a control group or an SC group. Both groups were fed the same basal diet but the SC group received SC supplementation (8 × 109 cfu/h/d through the ruminal fistula) following a two-period crossover design. Each period consisted of four phases, each of which lasted 17 d: 10 d for dietary adaptation, 6 d for degradation study, and 1 d for rumen sample collection. From the 1st to the 4th phase, steers were fed in a stepwise fashion with increasing CTFRs, i.e., 30:70, 50:50, 70:30, and 90:10. The kinetics of dry matter and fiber degradation of alfalfa pellets were evaluated; the rumen microbial populations were detected using real-time PCR. The results revealed no significant (P > 0.05) interactions between dietary CTFR and SC for most parameters. Dietary CTFR had a significant effect (P < 0.01) on degradation characteristics of alfalfa pellets and the copies of rumen microorganism; the increasing concentrate level resulted in linear, quadratic or cubic variation trend for these parameters. SC supplementation significantly (P < 0.05) affected dry matter (DM) and neutral detergent fiber (NDF) degradation rates (cDM, cNDF) and NDF effective degradability (EDNDF). Compared with the control group, there was an increasing trend of rumen fungi and protozoa in SC group (P < 0.1); copies of total bacteria in SC group were significantly higher (P < 0.05). Additionally, percentage of Ruminobacter amylophilus was significantly lower (P < 0.05) but percentage of Selenomonas ruminantium was significantly

  9. Increasing linseed supply in dairy cow diets based on hay or corn silage: Effect on enteric methane emission, rumen microbial fermentation, and digestion.

    PubMed

    Martin, C; Ferlay, A; Mosoni, P; Rochette, Y; Chilliard, Y; Doreau, M

    2016-05-01

    We investigated the effects of increasing extruded linseed supply in diets based on hay (H; experiment 1) or corn silage (CS; experiment 2) on enteric methane (CH4) emission, rumen microbial and fermentation parameters, and rumen and total-tract digestibility. In each experiment, 4 lactating Holstein cows fitted with cannulas at the rumen and proximal duodenum were used in a 4×4 Latin square design (28-d periods). Cows were fed ad libitum a diet [50:50 and 60:40 forage:concentrate on a dry matter (DM) basis for experiments 1 and 2, respectively] without supplementation (H0, CS0) or supplemented with extruded linseed at 5% (H5, CS5), 10% (H10, CS10), and 15% (H15, CS15) of dietary DM (i.e., 1.8, 3.6 and 5.4% total fatty acids added, respectively). All measurements were carried out during the last 8 d of each period. Linseed supply linearly decreased daily CH4 emission in cows fed H diets (from 486 to 289g/d for H0 to H15, on average) and CS diets (from 354 to 207g/d for CS0 to CS15, on average). The average decrease in CH4 per kilogram of DM intake was, respectively, -7, -15, and -38% for H5, H10, H15 compared with the H0 diet, and -4, -8, and -34% for CS5, CS10, and CS15 compared with the CS0 diet. The same dose-response effect was observed on CH4 emission in percent of gross energy intake, per kilogram of nutrient digested, and per kilogram of 4% fat- and 3.3% protein-corrected milk (FPCM) in both experiments. Changes in the composition of rumen volatile fatty acids in response to increasing linseed supply resulted in a moderate or marked linear decrease in acetate:propionate ratio for H or CS diets, respectively. The depressive effect of linseed on total protozoa concentration was linear for H diets (-15 to -40%, on average, for H5 to H15 compared with H0) and quadratic for CS diets (-17 to -83%, on average, for CS5 to CS15 compared with CS0). Concentration of methanogens was similar among H or CS diets. The energetic benefits from the decreased CH4 emission

  10. Precision-feeding dairy heifers a high rumen-degradable protein diet with different proportions of dietary fiber and forage-to-concentrate ratios.

    PubMed

    Lascano, G J; Koch, L E; Heinrichs, A J

    2016-09-01

    The objective of this experiment was to determine the effects of feeding a high-rumen-degradable protein (RDP) diet when dietary fiber content is manipulated within differing forage-to-concentrate ratio (F:C) on nutrient utilization of precision-fed dairy heifers. Six cannulated Holstein heifers (486.98±15.07kg of body weight) were randomly assigned to 2 F:C, low- (45% forage; LF) and high-forage (90% forage; HF) diets and to a fiber proportion sequence [33% grass hay and wheat straw (HS), 67% corn silage (CS; low fiber); 50% HS, 50% CS (medium fiber); and 67% HS, 33% CS (high fiber)] within forage proportion administered according to a split-plot, 3×3 Latin square design (16-d periods). Heifers fed LF had greater apparent total-tract organic matter digestibility coefficients (dC), neutral detergent fiber, and cellulose than those fed LC diets. Substituting CS with HS resulted in a linear reduction in dry matter, organic matter, and cellulose dC. Nitrogen dC was not different between F:C or with increasing proportions of HS in diets, but N retention tended to decrease linearly as HS was increased in the diets. Predicted microbial protein flow to the duodenum decreased linearly with HS addition and protozoa numbers HS interacted linearly, exhibiting a decrease as HS increased for LF, whereas no effects were observed for HF. Blood urea N increased linearly as HS was incorporated. The LF-fed heifers had a greater ruminal volatile fatty acids concentration. We noted a tendency for a greater dry matter, and a significantly higher liquid fraction turnover rate for HF diets. There was a linear numerical increase in the liquid and solid fraction turnover rate as fiber was added to the diets. Rumen fermentation parameters and fractional passages (solid and liquid) rates support the reduction in dC, N retention, and microbial protein synthesis observed as more dietary fiber is added to the rations of dairy heifers precision-fed a constant proportion of rumen

  11. Effects of variation of dietary sulfur on movement of sulfur in sheep rumen

    SciTech Connect

    Kandylis, K.; Bray, A.C.

    1987-01-01

    Effects of variations in dietary sulfur on rumen sulfur dynamics were studied under steady state conditions. In the first experimental period, three sheep were given 33.3 g of a pelleted diet hourly containing 1.59 g sulfur/kg (low) and in the second period the sulfur content was increased to 3.21 g/kg (high) by the addition of sodium sulfate. The daily sulfur intake was 1.158 g on the low sulfur diet and .545 g of this passed from the rumen in protein, .614 g was calculated to be absorbed from the rumen as sulfide, and .052 g was estimated to be recycled to the rumen. For sheep with daily intakes of 2.317 g sulfur, 1.212 g passed from the rumen in protein, 1.078 g was absorbed from the rumen, and .093 g was estimated to be recycled. It was estimated that 127 and 165 g microbial protein were synthesized/kg organic matter truly digested in the rumen for low and high sulfur diets, respectively. A simple model using simultaneous equations was proposed to describe rumen sulfur metabolism.

  12. Direct electricity recovery from Canna indica by an air-cathode microbial fuel cell inoculated with rumen microorganisms.

    PubMed

    Zang, Guo-Long; Sheng, Guo-Ping; Tong, Zhong-Hua; Liu, Xian-Wei; Teng, Shao-Xiang; Li, Wen-Wei; Yu, Han-Qing

    2010-04-01

    Aquatic plants are widely used for phytoremediation, and effective disposal methods should be pursued for their utilization and to avoid further environmental pollution problems. This study demonstrated that, using an air-cathode microbial fuel cell (MFC) inoculated with rumen microorganisms, electricity could be directly produced with a maximum power density of 0.405 W/m(3) from Canna indica (canna), a lignocellulosic aquatic plant rich in cellulose, hemicellulose, and lignin, without pretreatment. The mechanisms of the Canna indica degradation in the MFC were elucidated through analyzing the changes of canna structure and intermediates, that is, soluble sugars and volatile fatty acids (VFAs), in the electricity generation process. The results showed that lignin was partially removed and more cellulose became exposed on the sample surface during the electricity generation in the MFC. The electron transfer in this MFC was mainly completed through electron shuttling via self-produced mediators. This work presents an attempt to understand how complex substrates like aquatic plants are decomposed in an MFC during electricity generation. It might, hopefully, provide a promising way to utilize lignocellulosic biomass for energy generation.

  13. Establishment of cellulolytic bacteria and development of fermentative activities in the rumen of gnotobiotically-reared lambs receiving the microbial additive Saccharomyces cerevisiae CNCM I-1077.

    PubMed

    Chaucheyras-Durand, F; Fonty, G

    2001-01-01

    We studied the effects of a yeast additive used in ruminant nutrition on the establishment of cellulolytic bacteria, on plant cell wall degradation and on digestive functions in the rumen of gnotobiotically-reared lambs. Cellulolytic bacteria inoculated to the lambs tended to become established earlier in the presence of Saccharomyces cerevisiae CNCM I-1077 (SC). In addition, their population was maintained at a higher level, when the physico-chemical conditions of the biotope were altered. In these lambs, specific activities of fibrolytic enzymes were greater, and in sacco degradation of wheat straw tended to increase. In the presence of SC there was a decrease in ruminal ammonia concentration and a higher volatile fatty acid (VFA) concentration when lambs were 20 to 50 days old. These data suggest that this yeast strain may stimulate the development of cellulolytic microflora and enhance microbial activity in the rumen of young ruminants. Such activity could be beneficial in preventing microbial imbalance and a reduction of rumen function efficiency in the case of nutritional transitions. Further studies with conventional animals will soon be performed in order to verify these dings.

  14. Rumen microbial and fermentation characteristics are affected differently by bacterial probiotic supplementation during induced lactic and subacute acidosis in sheep

    PubMed Central

    2012-01-01

    Background Ruminal disbiosis induced by feeding is the cause of ruminal acidosis, a digestive disorder prevalent in high-producing ruminants. Because probiotic microorganisms can modulate the gastrointestinal microbiota, propionibacteria- and lactobacilli-based probiotics were tested for their effectiveness in preventing different forms of acidosis. Results Lactic acidosis, butyric and propionic subacute ruminal acidosis (SARA) were induced by feed chalenges in three groups of four wethers intraruminally dosed with wheat, corn or beet pulp. In each group, wethers were either not supplemented (C) or supplemented with Propionibacterium P63 alone (P) or combined with L. plantarum (Lp + P) or L. rhamnosus (Lr + P). Compared with C, all the probiotics stimulated lactobacilli proliferation, which reached up to 25% of total bacteria during wheat-induced lactic acidosis. This induced a large increase in lactate concentration, which decreased ruminal pH. During the corn-induced butyric SARA, Lp + P decreased Prevotella spp. proportion with a concomitant decrease in microbial amylase activity and total volatile fatty acids concentration, and an increase in xylanase activity and pH. Relative to the beet pulp-induced propionic SARA, P and Lr + P improved ruminal pH without affecting the microbial or fermentation characteristics. Regardless of acidosis type, denaturing gradient gel electrophoresis revealed that probiotic supplementations modified the bacterial community structure. Conclusion This work showed that the effectiveness of the bacterial probiotics tested depended on the acidosis type. Although these probiotics were ineffective in lactic acidosis because of a deeply disturbed rumen microbiota, some of the probiotics tested may be useful to minimize the occurrence of butyric and propionic SARA in sheep. However, their modes of action need to be further investigated. PMID:22812531

  15. Effect of niacin supplementation on rumen fermentation characteristics and nutrient flow at the duodenum in lactating dairy cows fed a diet with a negative rumen nitrogen balance.

    PubMed

    Aschemann, Martina; Lebzien, Peter; Hüther, Liane; Südekum, Karl-Heinz; Dänicke, Sven

    2012-08-01

    The aim of the present experiment was to ascertain if a daily niacin supplementation of 6 g/cow to lactating dairy cow diets can compensate for the decrease in rumen microbial fermentation due to a negative rumen nitrogen balance (RNB). A total of nine ruminally and duodenally fistulated lactating multiparous German Holstein cows was used. The diets consisted of 10 kg dry matter (DM) maize silage and 7 kg DM concentrate and differed as follows: (i) Diet RNB- (n = 6) with energy and utilisable crude protein (CP) at the duodenum (uCP) according to the average requirement of the animals, but with a negative RNB (-0.41 g N/MJ metabolisable energy [ME]); (ii) Diet RNB0 (n = 7) with energy, uCP, and RNB (0.08 g N/MJ ME) according to the average requirement of the animals; and (iii) Diet NA (nicotinic acid; n = 5), which was the same diet as RNB-, but supplemented with 6 g niacin/d. The negative RNB affected the rumen fermentation pattern and reduced ammonia content in rumen fluid and the daily duodenal flows of microbial CP (MP) and uCP. Niacin supplementation increased the apparent ruminal digestibility of neutral detergent fibre. The efficiency of microbial protein synthesis per unit of rumen degradable CP was higher, whereby the amount of MP reaching the duodenum was unaffected by niacin supplementation. The number of protozoa in rumen fluid was higher in NA treatment. The results indicated a more efficient use of rumen degradable N due to changes in the microbial population in the rumen when niacin was supplemented to diets deficient in RNB for lactating dairy cows. PMID:22924176

  16. Effect of niacin supplementation on rumen fermentation characteristics and nutrient flow at the duodenum in lactating dairy cows fed a diet with a negative rumen nitrogen balance.

    PubMed

    Aschemann, Martina; Lebzien, Peter; Hüther, Liane; Südekum, Karl-Heinz; Dänicke, Sven

    2012-08-01

    The aim of the present experiment was to ascertain if a daily niacin supplementation of 6 g/cow to lactating dairy cow diets can compensate for the decrease in rumen microbial fermentation due to a negative rumen nitrogen balance (RNB). A total of nine ruminally and duodenally fistulated lactating multiparous German Holstein cows was used. The diets consisted of 10 kg dry matter (DM) maize silage and 7 kg DM concentrate and differed as follows: (i) Diet RNB- (n = 6) with energy and utilisable crude protein (CP) at the duodenum (uCP) according to the average requirement of the animals, but with a negative RNB (-0.41 g N/MJ metabolisable energy [ME]); (ii) Diet RNB0 (n = 7) with energy, uCP, and RNB (0.08 g N/MJ ME) according to the average requirement of the animals; and (iii) Diet NA (nicotinic acid; n = 5), which was the same diet as RNB-, but supplemented with 6 g niacin/d. The negative RNB affected the rumen fermentation pattern and reduced ammonia content in rumen fluid and the daily duodenal flows of microbial CP (MP) and uCP. Niacin supplementation increased the apparent ruminal digestibility of neutral detergent fibre. The efficiency of microbial protein synthesis per unit of rumen degradable CP was higher, whereby the amount of MP reaching the duodenum was unaffected by niacin supplementation. The number of protozoa in rumen fluid was higher in NA treatment. The results indicated a more efficient use of rumen degradable N due to changes in the microbial population in the rumen when niacin was supplemented to diets deficient in RNB for lactating dairy cows.

  17. Rumen dry matter and crude protein degradability of extracted or untreated oilseeds and Leucaena leucocephala leaves.

    PubMed

    Gralak, M A; Kamalu, T; von Keyserlingk, M A; Kulasek, G W

    1997-01-01

    A study was undertaken to determine the rumen DM and CP degradability characteristics of soyabean, canola seed, peanut, palm kernel and Leucaena leucocephala leaves. The oilseeds were either treated with n-hexane to extract the fat or left untreated. Nylon bags were incubated in each of four rumen cannulated sheep for 0, 2, 4, 6, 12, 24 and 48 h. Animals were fed on a diet consisting of meadow hay (ad libitum) and 150 g of concentrate twice daily. Fat extraction caused a decrease (P < or = 0.05) in DM disappearance of soyabean at 0, 2, 4, 6 and 12 h and of peanuts at all incubation times. CP disappearance from peanuts was reduced (P < or = 0.05) as a result of fat extraction at 0, 2, 4, 6 and 12 h. Fat extraction of canola seed increased CP disappearance at 0, 2, 4, 6 and 24 h (P < or = 0.05). However, in the case of defatted canola seed, an increase in DM disappearance (P < or = 0.05) was observed in the first 4 incubation times and a decrease (P < or = 0.05) in the later times. Fat extraction increased (P < or = 0.05) DM disappearance of palm kernel at 0 and 48 h, but reduced it at 4, 6 and 24 h. CP disappearance of palm kernel was improved by treatment (P < or = 0.05) at 0, 4, 24 and 48 h and decreased at 12 h. In the case of palm kernel the largest differences in DM and CP disappearance occurred between the 24 and 48 h incubation times. Degradability characteristics for DM and CP of full-fat soyabean, canola seed and peanut were comparable to those of the full fat samples. Effective DM degradability of soyabean, canola seed and peanuts was 72.2 and 71.9; 74.1 and 66.8; and 85.9 and 70.8 for full fat and extracted feeds, respectively. Effective CP degradability was similar in all oilseeds with the exception of the extracted canola seed. Therefore, the incorporation of full-fat soyabean, canola seed and peanut into ruminant rations can be considered as a means of increasing the energy balance. Both palm kernel DM and CP degradabilities were characterized by slow

  18. Comparative Survey of Rumen Microbial Communities and Metabolites across One Caprine and Three Bovine Groups, Using Bar-Coded Pyrosequencing and 1H Nuclear Magnetic Resonance Spectroscopy

    PubMed Central

    Lee, Hyo Jung; Jung, Ji Young; Oh, Young Kyoon; Lee, Sang-Suk; Madsen, Eugene L.

    2012-01-01

    Pyrosequencing of 16S rRNA genes (targeting Bacteria and Archaea) and 1H nuclear magnetic resonance were applied to investigate the rumen microbiota and metabolites of Hanwoo steers in the growth stage (HGS), Hanwoo steers in the late fattening stage (HFS), Holstein-Friesian dairy cattle (HDC), and Korean native goats (KNG) in the late fattening stage. This was a two-part investigation. We began by comparing metabolites and microbiota of Hanwoo steers at two stages of husbandry. Statistical comparisons of metabolites and microbial communities showed no significant differences between HFS and HGS (differing by a dietary shift at 24 months and age [67 months versus 12 months]). We then augmented the study by extending the investigation to HDC and KNG. Overall, pyrosequencing of 16S rRNA genes showed that the rumens had highly diverse microbial communities containing many previously undescribed microorganisms. Bioinformatic analysis revealed that the bacterial sequences were predominantly affiliated with four phyla—Bacteroidetes, Firmicutes, Fibrobacteres, and Proteobacteria—in all ruminants. However, interestingly, the bacterial reads belonging to Fibrobacteres were present at a very low abundance (<0.1%) in KNG. Archaeal community analysis showed that almost all of these reads fell into a clade related to, but distinct from, known cultivated methanogens. Statistical analyses showed that the microbial communities and metabolites of KNG were clearly distinct from those of other ruminants. In addition, bacterial communities and metabolite profiles of HGS and HDC, fed similar diets, were distinctive. Our data indicate that bovine host breeds override diet as the key factor that determines bacterial community and metabolite profiles in the rumen. PMID:22706048

  19. Comparative survey of rumen microbial communities and metabolites across one caprine and three bovine groups, using bar-coded pyrosequencing and ¹H nuclear magnetic resonance spectroscopy.

    PubMed

    Lee, Hyo Jung; Jung, Ji Young; Oh, Young Kyoon; Lee, Sang-Suk; Madsen, Eugene L; Jeon, Che Ok

    2012-09-01

    Pyrosequencing of 16S rRNA genes (targeting Bacteria and Archaea) and (1)H nuclear magnetic resonance were applied to investigate the rumen microbiota and metabolites of Hanwoo steers in the growth stage (HGS), Hanwoo steers in the late fattening stage (HFS), Holstein-Friesian dairy cattle (HDC), and Korean native goats (KNG) in the late fattening stage. This was a two-part investigation. We began by comparing metabolites and microbiota of Hanwoo steers at two stages of husbandry. Statistical comparisons of metabolites and microbial communities showed no significant differences between HFS and HGS (differing by a dietary shift at 24 months and age [67 months versus 12 months]). We then augmented the study by extending the investigation to HDC and KNG. Overall, pyrosequencing of 16S rRNA genes showed that the rumens had highly diverse microbial communities containing many previously undescribed microorganisms. Bioinformatic analysis revealed that the bacterial sequences were predominantly affiliated with four phyla-Bacteroidetes, Firmicutes, Fibrobacteres, and Proteobacteria-in all ruminants. However, interestingly, the bacterial reads belonging to Fibrobacteres were present at a very low abundance (<0.1%) in KNG. Archaeal community analysis showed that almost all of these reads fell into a clade related to, but distinct from, known cultivated methanogens. Statistical analyses showed that the microbial communities and metabolites of KNG were clearly distinct from those of other ruminants. In addition, bacterial communities and metabolite profiles of HGS and HDC, fed similar diets, were distinctive. Our data indicate that bovine host breeds override diet as the key factor that determines bacterial community and metabolite profiles in the rumen.

  20. Effects of Protein Level and Mangosteen Peel Pellets (Mago-pel) in Concentrate Diets on Rumen Fermentation and Milk Production in Lactating Dairy Crossbreds

    PubMed Central

    Norrapoke, T.; Wanapat, M.; Wanapat, S.

    2012-01-01

    Four, lactating dairy crossbreds (50%×50% Holstein Friesian×Native Zebu cattle) were randomly assigned according to a 2×2 factorial arrangement (two protein levels and two levels of mangosteen peel pellets (Mago-pel)) in a 4×4 Latin square design to receive four dietary treatments. All cows received concentrate at a proportion of 1 kg concentrate per 2 kg of milk yield, and urea-treated 5% rice straw (UTRS) was given ad libitum. It was found that total dry matter intakes, nutrient digestibility, ruminal pH and NH3-N concentrations were not affected (p>0.05) by treatments. Concentrations of ruminal pH and NH3-N were not affected by dietary treatments although the concentration of BUN varied significantly (p<0.05) between protein levels (p<0.05). The populations of rumen bacteria and fungal zoospores did not differ among treatments (p>0.05); however, the population of protozoa was decreased (p<0.05) when cows received Mago-pel supplementation. The composition of the population of bacteria, identified by real-time PCR technique, including total bacteria, methanogens, Fibrobacter succinogenes and Ruminococcus albus was similar (p>0.05) among dietary treatments (p>0.05); however, copy numbers of Ruminococcus flavefaciens was increased when protein level increased (p<0.05). Microbial protein synthesis, in terms of both quantity and efficiency, was enriched by Mago-pel supplementation. Milk yield was greatest in cows fed UTRS based diets with concentrate containing protein at 16% CP with Mago-pel, but were lowest without Mago-pel (p<0.05). In addition, protein level and supplementation of Mago-pel did not affect (p>0.05) milk composition except solids-not-fat which was higher in cows fed the diet with 19% CP. Therefore, feeding a concentrate containing 16% CP together with 300 g/hd/d Mago-pel supplementation results in changes in rumen fermentation and microbial population and improvements in milk production in lactating dairy crossbreds fed on UTRS. PMID:25049652

  1. Tuning microbial hosts for membrane protein production

    PubMed Central

    2009-01-01

    The last four years have brought exciting progress in membrane protein research. Finally those many efforts that have been put into expression of eukaryotic membrane proteins are coming to fruition and enable to solve an ever-growing number of high resolution structures. In the past, many skilful optimization steps were required to achieve sufficient expression of functional membrane proteins. Optimization was performed individually for every membrane protein, but provided insight about commonly encountered bottlenecks and, more importantly, general guidelines how to alleviate cellular limitations during microbial membrane protein expression. Lately, system-wide analyses are emerging as powerful means to decipher cellular bottlenecks during heterologous protein production and their use in microbial membrane protein expression has grown in popularity during the past months. This review covers the most prominent solutions and pitfalls in expression of eukaryotic membrane proteins using microbial hosts (prokaryotes, yeasts), highlights skilful applications of our basic understanding to improve membrane protein production. Omics technologies provide new concepts to engineer microbial hosts for membrane protein production. PMID:20040113

  2. Removal of digesta components from the rumen of steers determined by sieving techniques and fluid, particulate and microbial markers.

    PubMed

    Dixon, R M; Milligan, L P

    1985-03-01

    When 103Ru-labelled Tris (1,10-phenanthroline) ruthenium II chloride (103Ru-P) particulate marker in aqueous solution was added to the rumen of four steers given 5.5 kg grass hay/d at two-hourly intervals, the distribution of 103Ru-P marker among rumen particles of various sizes was the same at 4 h, 3 d and 7 d after administration, the concentration of 103Ru-P/g dry matter (DM) was inversely related to particle size and 0.30 of the 103Ru-P was associated with the DM of particles too large to be moved from the rumen at a meaningful rate. Thus, fractional outflow rate (FOR) of 103Ru-P would reflect, but was not a direct measure of, the FOR of the small particle pool in the rumen. When rumen digesta were labelled with 103Ru-P, placed in nylon cloth bags and incubated in vitro with unlabelled digesta, 59% of the 103Ru-P disappeared from the nylon bag in 24 h, and 74% in 48 h. Similar results were obtained when large particles (retained by a 3.2 mm mesh screen during wet sieving) from rumen digesta were subjected to this procedure. In a further experiment, the steers were given the hay in either the long or ground form and drinking water to which 10 g sodium chloride/l were, or were not, added. The FOR of 51CrEDTA in centrifuged rumen fluid was increased (P less than 0.05) from 1.78 to 2.10/d by grinding of the hay diet, but was not influenced by the intake of an additional 257 g NaCl/d. The FOR values of 103Ru-P in mixed rumen digesta and organic 35S in micro-organisms were linearly correlated (P less than 0.05) and were not affected (P greater than 0.05) by grinding and salt treatments. On average, the FOR of organic 35S in micro-organisms was 0.41 of that of 51CrEDTA in centrifuged rumen fluid and 0.85 of that of 103Ru-P in rumen digesta respectively. Grinding of the hay did not (P greater than 0.05) change the proportion of rumen DM (0.476-0.515) or faecal DM (0.107-0.153) retained by the 3.2 mm mesh and larger screens. FOR from the rumen of a given size group of

  3. The rumen plasmidome

    PubMed Central

    Mizrahi, Itzhak

    2012-01-01

    Plasmids are episomally replicating genetic elements which carry backbone genes that are important for their replication and maintenance within their host, and accessory genes that might confer an advantage to their host in its ecological niche. As such, they are often perceived as a powerful evolutionary force, which horizontally introduces new traits into bacterial cells and genomes. In our recent publication “Insight into the rumen plasmidome” we characterized the metagenomic plasmid population of the bovine rumen microbial ecological niche. The rumen is the first compartment of the digestive tract of ruminants; it functions as a pre-gastric anaerobic fermentation chamber, where plant fibers are degraded and converted into chemical compounds which are subsequently absorbed and digested by the animal. PMID:23061023

  4. Supplementation of cattle fed tropical grasses with microalgae increases microbial protein production and average daily gain.

    PubMed

    Costa, D F A; Quigley, S P; Isherwood, P; McLennan, S R; Poppi, D P

    2016-05-01

    A series of 3 experiments were conducted to evaluate the use of microalgae as supplements for ruminants consuming low-CP tropical grasses. In Exp. 1, the chemical composition and in vitro protein degradability of 9 algae species and 4 protein supplements were determined. In Exp. 2, rumen function and microbial protein (MCP) production were determined in steers fed speargrass hay alone or supplemented with , , , or cottonseed meal (CSM). In Exp. 3, DMI and ADG were determined in steers fed speargrass hay alone or supplemented with increasing amounts of NPN (urea combined with ammonia sulfate), CSM, or . In Exp. 1, the CP content of and (675 and 580 g/kg DM) was highest among the algae species and higher than the other protein supplements evaluated, and sp. had the highest crude lipid (CL) content (198 g/kg DM). In Exp. 2, supplementation increased speargrass hay intake, the efficiency of MCP production, the fractional outflow rate of digesta from the rumen, the concentration of NHN, and the molar proportion of branched-chain fatty acids in the rumen fluid of steers above all other treatments. acceptance by steers was low and this resulted in no significant difference to unsupplemented steers for all parameters measured for this algae supplement. In Exp. 3, ADG linearly increased with increasing supplementary N intake from both and NPN, with no difference between the 2 supplements. In contrast, ADG quadratically increased with increasing supplementary N intake from CSM. It was concluded that and may potentially be used as protein sources for cattle grazing low-CP pastures. PMID:27285702

  5. Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail

    PubMed Central

    2012-01-01

    Background A complete saccharification of plant polymers is the critical step in the efficient production of bio-alcohols. Beta-glucosidases acting in the degradation of intermediate gluco-oligosaccharides produced by cellulases limit the yield of the final product. Results In the present work, we have identified and then successfully cloned, expressed, purified and characterised 4 highly active beta-glucosidases from fibre-adherent microbial community from the cow rumen. The enzymes were most active at temperatures 45–55°C and pH 4.0-7.0 and exhibited high affinity and activity towards synthetic substrates such as p-nitrophenyl-beta-D-glucopyranoside (pNPbetaG) and pNP-beta-cellobiose, as well as to natural cello-oligosaccharides ranging from cellobiose to cellopentaose. The apparent capability of the most active beta-glucosidase, herein named LAB25g2, was tested for its ability to improve, at low dosage (31.25 units g-1 dry biomass, using pNPbetaG as substrate), the hydrolysis of pre-treated corn stover (dry matter content of 20%; 350 g glucan kg-1 dry biomass) in combination with a beta-glucosidase-deficient commercial Trichoderma reseei cellulase cocktail (5 units g-1 dry biomass in the basis of pNPbetaG). LAB25g2 increased the final hydrolysis yield by a factor of 20% (44.5 ± 1.7% vs. 34.5 ± 1.5% in control conditions) after 96–120 h as compared to control reactions in its absence or in the presence of other commercial beta-glucosidase preparations. The high stability (half-life higher than 5 days at 50°C and pH 5.2) and 2–38000 fold higher (as compared with reported beta-glucosidases) activity towards cello-oligosaccharides may account for its performance in supplementation assays. Conclusions The results suggest that beta-glucosidases from yet uncultured bacteria from animal digestomes may be of a potential interest for biotechnological processes related to the effective bio-ethanol production in combination with low dosage of commercial cellulases

  6. Dynamics of protozoa in the rumen of sheep.

    PubMed

    Leng, R A

    1982-09-01

    as 32% of the total microbial protein synthesis in the rumen but that 65% of the protozoa die and are degraded in the rumen.

  7. Rumen degradability characteristics of normal maize stover and silage, and quality protein maize silage-based diets offered to cows.

    PubMed

    Tamir, Berhan; Gebrehawariat, Ephrem; Tegegne, Azage; Kortu, Mohammed Y

    2012-10-01

    Rumen degradability characteristics of dry matter (DM), organic matter (OM) and crude protein (CP) of normal maize (NM) stover (T1)-, NM silage (T2)- and quality protein maize (QPM) silage (T3)-based diets were studied using three rumen-fistulated Boran × Friesian non-lactating cows (371 ± 32.00 kg) in 3 × 3 Latin Square Design. Cows were supplemented with a similar concentrate mix. In sacco degradability of DM and OM indicated that the (a) values of DM (128) and OM (114) for NM stover were lower (P < 0.001) than that for NM silage (268 and 253) and for QPM silage (323 and 303), respectively. The (a) value for CP was lower (P < 0.05) for QPM silage (286) than for NM stover (404) and NM silage (326). The (b) values of DM in NM stover (597) and NM silage (535) were higher (P < 0.05) than in QPM silage (499). The (b) value of CP in NM stover (372) was lower (P < 0.05) than in NM silage (655) and in QPM silage (608). Rate of degradation of OM in NM stover and NM silage, each with 0.03, was faster (P < 0.01) than in QPM silage (0.02). Moreover, QPM silage had higher potentially degradable fraction for DM (821) (P < 0.05) and OM (840) (P < 0.01) than DM (725) and OM (712) in NM stover. The mean rumen ammonia concentration (209 mg/l) of QPM silage was higher (P < 0.05) than that of NM stover (179 mg/l) and NM silage (170 mg/l). The average rumen pH (6.1) in cows fed QPM silage was lowest (P < 0.05) compared to pH (6.3) in cows fed either NM stover or silage. The concentration of total volatile fatty acids (116 mmol/l) in the rumen of cows incubated with QPM silage was higher (P < 0.001) than in those incubated with NM stover (113 mmol/l) and NM silage (110 mmol/l). It was concluded that QPM silage-based diet was superior in DM and OM degradability, and had higher ammonia and VFA concentration than NM stover-based diet. No differences have been observed in all parameters measured between QPM and NM silages. PMID:22366928

  8. Rumen Metagenomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The rumen microbiome plays a critical role in normal physiology and nutrition of ruminants. Alterations in the rumen microbiome have important physiological and pathological implications. The advent of next-generation sequencing technologies and rapid development of computational tools and reference...

  9. Supplementing barley or rapeseed meal to dairy cows fed grass-red clover silage: I. Rumen degradability and microbial flow.

    PubMed

    Ahvenjärvi, S; Vanhatalo, A; Huhtanen, P

    2002-08-01

    The present study was conducted to measure the flow of microbial and nonmicrobial N fractions entering the omasal canal of lactating dairy cows fed grass-red clover silage supplemented with barley and rapeseed meal. Four ruminally cannulated Finnish Ayrshire dairy cows were fed, in a 4 x 4 Latin square design, grass-red clover silage alone or supplemented with (on DM basis) 5.1 kg/d of barley, 1.9 kg/d of rape-seed meal or 5.1 kg/d of barley and 1.9 kg/d rapeseed meal. Nonammonia N flow entering the omasal canal was fractionated into microbial and nonmicrobial N using 15N. Microbial N was fractionated into N associated with liquid-associated bacteria, particle-associated bacteria, and protozoa. Supplementation of diets with barley increased microbial N flow entering the omasal canal (P < 0.01) but had no effect on nonmicrobial N flow. Increased microbial N flow was attributed to liquid-associated bacteria and protozoa. Barley had no effect on apparent ruminal N degradability, but increased true ruminal N degradability (P < 0.01). Barley had no effect on urinary N excretion, but increased daily N retention (P = 0.03). Furthermore, barley supplementation decreased ruminal (P = 0.02) and total tract (P < 0.01) NDF digestibility. Supplementation of diets with rapeseed meal increased apparent ruminal N degradability (P < 0.01) and nonmicrobial N flow entering the omasal canal (P < 0.01), but had no effect on true ruminal N degradability. Despite higher N excretion in urine, rapeseed meal improved daily N retention (P < 0.01). Milk yield was increased (P < 0.01) by barley and rapeseed meal supplements, with the responses being additive. Responses attained with barley were primarily due to increased energy supply for ruminal microbes and improvements in energy and protein supply for the animal. However, provision of readily digestible carbohydrates in barley did not improve microbial capture of ruminal ammonia. Benefits associated with rapeseed meal supplementation were

  10. Effects of Adaptation of In vitro Rumen Culture to Garlic Oil, Nitrate, and Saponin and Their Combinations on Methanogenesis, Fermentation, and Abundances and Diversity of Microbial Populations

    PubMed Central

    Patra, Amlan K.; Yu, Zhongtang

    2015-01-01

    This study investigated the effects of garlic oil (0.25 g/L), nitrate (5 mM), and quillaja saponin (0.6 g/L), alone and in binary or ternary combinations, on methanogenesis, rumen fermentation, and abundances of select microbial populations using in vitro rumen cultures. Potential adaptation to these compounds was also examined by repeated transfers of the cultures on alternate days until day 18. All treatments except saponin alone significantly decreased methanogenesis. Ternary combinations of garlic oil, nitrate, and saponin additively/synergistically suppressed methane production by 65% at day 2 and by 40% at day 18. Feed digestion was not adversely affected by any of the treatments at day 2, but was decreased by the combinations (binary and ternary) of garlic oil with the other inhibitors at days 10 and 18. Saponin, alone or in combinations, and garlic oil alone lowered ammonia concentration at day 2, while nitrate increased ammonia concentration at days 10 and 18. Total volatile fatty acid concentration was decreased by garlic oil alone or garlic oil-saponin combination. Molar proportions of acetate and propionate were affected to different extents by the different treatments. The abundances of methanogens were similar among treatments at day 2; however, garlic oil and its combination with saponin and/or nitrate at day 10 and all treatments except saponin at day 18 significantly decreased the abundances of methanogens. All the inhibitors, either alone or in combinations, did not adversely affect the abundances of total bacteria or Ruminococcus flavefaciens. However, at day 18 the abundances of Fibrobacter succinogenes and Ruminococcus albus were lowered in the presence of garlic oil and saponin, respectively. The results suggest that garlic oil-nitrate-saponin combination (at the doses used in this study) can effectively decreases methanogenesis in the rumen, but its efficacy may decrease while inhibition to feed digestion can increase over time. PMID:26733975

  11. Changes of Microbial Population in the Rumen of Dairy Steers as Influenced by Plant Containing Tannins and Saponins and Roughage to Concentrate Ratio

    PubMed Central

    Anantasook, N.; Wanapat, M.; Cherdthong, A.; Gunun, P.

    2013-01-01

    The objective of this study was to investigate microbial population in the rumen of dairy steers as influenced by supplementing with dietary condensed tannins and saponins and different roughage to concentrate ratios. Four, rumen fistulated dairy steers (Bos indicus) were used in a 2×2 factorial arrangement in a 4×4 Latin square design. The main factors were two roughage to concentrate ratios (R:C, 60:40 and 40:60) and two supplementations of rain tree pod meal (RPM) (0 and 60 g/kg of total DM intake). Chopped 30 g/kg urea treated rice straw was used as a roughage source. All animals received feed according to respective R:C ratios at 25 g/kg body weight. The RPM contained crude tannins and saponins at 84 and 143 g/kg of DM, respectively. It was found that ruminal pH decreased while ruminal temperature increased by a higher concentrate ratio (R:C 40:60) (p<0.05). In contrast, total bacterial, Ruminococus albus and viable proteolytic bacteria were not affected by dietary supplementation. Numbers of fungi, cellulolytic bacteria, Fibrobactor succinogenes and Ruminococus flavefaciens were higher while amylolytic bacteria was lower when steers were fed at 400 g/kg of concentrate. The population of Fibrobactor succinogenes, was found to be higher with RPM supplementation. In addition, the use of real-time PCR technique indicated that the population of protozoa and methanogens were decreased (p<0.05) with supplementation of RPM and with an increasing concentrate ratio. Supplementation of RPM and feeding different concentrate ratios resulted in changing the rumen microbes especially, when the animals were fed at 600 g/kg of concentrate and supplemented with RPM which significantly reduced the protozoa and methanogens population. PMID:25049745

  12. Effects of replacing dietary starch with neutral detergent-soluble fibre on ruminal fermentation, microbial synthesis and populations of ruminal cellulolytic bacteria using the rumen simulation technique (RUSITEC).

    PubMed

    Zhao, X H; Liu, C J; Liu, Y; Li, C Y; Yao, J H

    2013-12-01

    A rumen simulation technique (RUSITEC) apparatus with eight 800 ml fermenters was used to investigate the effects of replacing dietary starch with neutral detergent-soluble fibre (NDSF) by inclusion of sugar beet pulp in diets on ruminal fermentation, microbial synthesis and populations of ruminal cellulolytic bacteria. Experimental diets contained 12.7, 16.4, 20.1 or 23.8% NDSF substituted for starch on a dry matter basis. The experiment was conducted over two independent 15-day incubation periods with the last 8 days used for data collection. There was a tendency that 16.4% NDSF in the diet increased the apparent disappearance of organic matter (OM) and neutral detergent fibre (NDF). Increasing dietary NDSF level increased carboxymethylcellulase and xylanase activity in the solid fraction and apparent disappearance of acid detergent fibre (ADF) but reduced the 16S rDNA copy numbers of Ruminococcus albus in both liquid and solid fractions and R. flavefaciens in the solid fraction. The apparent disappearance of dietary nitrogen (N) was reduced by 29.6% with increased dietary NDSF. Substituting NDSF for starch appeared to increase the ratios of acetate/propionate and methane/volatile fatty acids (VFA) (mol/mol). Replacing dietary starch with NDSF reduced the daily production of ammonia-N and increased the growth of the solid-associated microbial pellets (SAM). Total microbial N flow and efficiency of microbial synthesis (EMS), expressed as g microbial N/kg OM fermented, tended to increase with increased dietary NDSF, but the numerical increase did not continue as dietary NDSF exceeded 20.1% of diet DM. Results suggested that substituting NDSF for starch up to 16.4% of diet DM increased digestion of nutrients (except for N) and microbial synthesis, and further increases (from 16.4% to 23.8%) in dietary NDSF did not repress microbial synthesis but did significantly reduce digestion of dietary N.

  13. Effect of the Ratio of Non-fibrous Carbohydrates to Neutral Detergent Fiber and Protein Structure on Intake, Digestibility, Rumen Fermentation, and Nitrogen Metabolism in Lambs

    PubMed Central

    Ma, T.; Tu, Y.; Zhang, N. F.; Deng, K. D.; Diao, Q. Y.

    2015-01-01

    This study aimed to investigate the effect of the ratio of non-fibrous carbohydrates to neutral detergent fibre (NFC/NDF) and undegraded dietary protein (UDP) on rumen fermentation and nitrogen metabolism in lambs. Four Dorper×thin-tailed Han crossbred lambs, averaging 62.3±1.9 kg of body weight and 10 mo of age, were randomly assigned to four dietary treatments of combinations of two levels of NFC/NDF (1.0 and 1.7) and two levels of UDP (35% and 50% of crude protein [CP]). Duodenal nutrient flows were measured with dual markers of Yb and Co, and microbial N (MN) synthesis was estimated using 15N. High UDP decreased organic matter (OM) intake (p = 0.002) and CP intake (p = 0.005). Ruminal pH (p<0.001), ammonia nitrogen (NH3-N; p = 0.008), and total volatile fatty acids (p<0.001) were affected by dietary NFC/NDF. The ruminal concentration of NH3-N was also affected by UDP (p<0.001). The duodenal flow of total MN (p = 0.007) was greater for lambs fed the high NFC/NDF diet. The amount of metabolisable N increased with increasing dietary NFC:NDF (p = 0.02) or UDP (p = 0.04). In conclusion, the diets with high NFC/NDF (1.7) and UDP (50% of CP) improved metabolisable N supply to lambs. PMID:26323398

  14. The large bowel--a supplementary rumen?

    PubMed

    Argenzio, R A; Stevens, C E

    1984-01-01

    The rumen and the mammalian large intestine are similar in many respects. Microbial protein appears to be synthesized and degraded in the digesta of both organs in a comparable manner. The VFA end-products of carbohydrate fermentation are produced in similar concentrations. Digesta pH is maintained with buffer added by the saliva or ileal fluid, HCO3 released into the lumen and rapid absorption of the organic acids. VFA are absorbed at equivalent rates by rumen epithelium and large intestinal mucosa. Over-production of VFA produces similar adverse effects. There is a considerable amount of species variation in the relative length and volume as well as the extent of sacculation of the large intestine. The caecum is the primary site for retention of digesta and microbial fermentation in the large intestine of rabbits, rodents and a few other species. However, the proximal colon is the major site of retention and fermentation in most mammals. Absorptions of Na and VFA appear to account for absorption of most of the water removed during passage of digesta through the large intestine. A relatively slow rate of Na absorption and release of HCO3 appears to provide the fluid and buffering capacity needed for efficient microbial digestion in the rumen and in the large intestine of some species. A more rapid absorption of Na by the large intestine of other species would aid in the conservation of Na and water. The many similarities between the large intestine and the rumen suggest that further comparison can provide additional information on both the function and diseases of these two organs. The rumen has proved to be accessible to a variety of procedures useful for the study of microbial digestive processes and its epithelium has provided a non-glandular tissue for studies of inorganic ion transport as well as the transport and metabolism of VFA. Comparative studies of the large intestine also can provide a better understanding of the functions and malfunctions of the

  15. Biohydrogenation of dietary n-3 PUFA and stability of ingested vitamin E in the rumen, and their effects on microbial activity in sheep.

    PubMed

    Chikunya, S; Demirel, G; Enser, M; Wood, J D; Wilkinson, R G; Sinclair, L A

    2004-04-01

    The present study investigated the susceptibility of dietary n-3 PUFA to ruminal biohydrogenation, the stability of ingested vitamin E in the rumen and the subsequent uptake of PUFA and vitamin E into plasma. Six cannulated sheep were assigned to six diets over five 33 d periods, in an incomplete 6x5 Latin square. The diets, based on dried grass, were formulated to supply 50 g fatty acids/kg DM using three lipid sources: Megalac (calcium soap of palm fatty acid distillate; Volac Ltd, Royston, Herts., UK), linseed (formaldehyde-treated; Trouw Nutrition, Northwich, Ches., UK) and linseed-fish oil (formaldehyde-treated linseed+fish oil). The diets were supplemented with 100 or 500 mg alpha-tocopheryl acetate/kg DM. Fat source or level of vitamin E in the diet did not alter microbial activity in the rumen. Biohydrogenation of linoleic acid (18 : 3n-6; 85-90 %), linolenic acid (18 : 3n-3; 88-93 %), docosahexaenoic acid (22 : 6n-3; 91 %) and EPA (20 : 5n-3; 92 %) was extensive. Feeding formaldehyde-treated linseed elevated concentrations of 18 : 3n-3 in plasma, whilst 22 : 6n-3 and 20 : 5n-3 were only increased by feeding the linseed-fish oil blend. Duodenal recovery of ingested vitamin E was high (range 0.79-0.92 mg/mg fed). High dietary vitamin E was associated with increased plasma alpha-tocopherol (2.57 v. 1.46 microg/ml for 500 and 100 mg alpha-tocopheryl acetate/kg DM respectively), although all concentrations were low. Plasma vitamin E levels, however, tended to decrease as the type and quantity of PUFA in the diet increased. The present study illustrates that nutritionally beneficial PUFA in both fish and linseed oils are highly susceptible to biohydrogenation in the rumen. Although alpha-tocopheryl acetate resisted degradation in the rumen, plasma vitamin E status remained deficient to borderline, suggesting either that uptake may have been impaired or metabolism post-absorption increased.

  16. Comparison of three 15N methods to correct for microbial contamination when assessing in situ protein degradability of fresh forages.

    PubMed

    Kamoun, M; Ammar, H; Théwis, A; Beckers, Y; France, J; López, S

    2014-11-01

    The use of stable (15)N as a marker to determine microbial contamination in nylon bag incubation residues to estimate protein degradability was investigated. Three methods using (15)N were compared: (15)N-labeled forage (dilution method, LF), (15)N enrichment of rumen solids-associated bacteria (SAB), and (15)N enrichment of rumen liquid-associated bacteria (LAB). Herbage from forages differing in protein and fiber contents (early-cut Italian ryegrass, late-cut Italian ryegrass, and red clover) were freeze-dried and ground and then incubated in situ in the rumen of 3 steers for 3, 6, 12, 24, and 48 h using the nylon bag technique. The (15)N-labeled forages were obtained by fertilizing the plots where herbage was grown with (15)NH4 (15)NO3. Unlabeled forages (obtained from plots fertilized with NH4NO3) were incubated at the same time that ((15)NH4)2SO4 was continuously infused into the rumen of the steers, and then pellets of labeled SAB and LAB were isolated by differential centrifugation of samples of ruminal contents. The proportion of bacterial N in the incubation residues increased from 0.09 and 0.45 g bacterial N/g total N at 3 h of incubation to 0.37 and 0.85 g bacterial N/g total N at 48 h of incubation for early-cut and late-cut ryegrass, respectively. There were differences (P < 0.001) between uncorrected N degradability values and those corrected for microbial contamination with all of the methods. Apparent N degradability of the low-N, high-fiber forage (late-cut ryegrass) was 0.51, whereas the corrected values were 0.85, 0.84, and 0.77 for the LF, SAB, and LAB methods, respectively. With early-cut ryegrass and red clover, the differences between uncorrected and corrected values ranged between 6% and 13%, with small differences among the labeling methods. Generally, methods using labeled forage or labeled SAB and LAB provided similar corrected degradability values. The accuracy in estimating the extent of degradation of protein in the rumen from in situ

  17. Microbial thiamin metabolism in the rumen simulating fermenter (RUSITEC): the effect of acidogenic conditions, a high sulfur level and added thiamin.

    PubMed

    Alves de Oliveira, L; Jean-Blain, C; Komisarczuk-Bony, S; Durix, A; Durier, C

    1997-10-01

    The effects of acidogenic conditions, a high S level and the addition of thiamin on the rumen microbial metabolism of thiamin were investigated in vitro in a semi-continuous fermenter (RUSITEC), using a factorial design. Acidogenic conditions were obtained by simultaneously increasing the starch: cellulose ratio and the amount of solid substrate fed, and by decreasing the buffering capacity of the liquid phase of the fermenter. S in the form of sulfate was supplied at two levels, one corresponding to a control amount of S (2 g/kg dietary DM), the second to an excess (5 g/kg DM) which is sufficient to trigger cerebrocortical necrosis (CCN) when used in vivo. Acidogenic conditions decreased the pH of the fermenters, CH4 production and cellulose digestibility, increased the short-chain fatty acid production, but had no effect on thiamin production. The high S level enhanced the production of sulfide considerably, had no effect ont he microbial metabolism of energy and N, and decreased thiamin production (326 v. 266 nmol/d). The added thiamin was rapidly converted into phosphorylated compounds which largely decreased the apparent synthesis of this vitamin by the rumen microflora. The total thiamin flow was increased by added thiamin. In no case was thiaminase activity in the fermenter liquid phase significantly modified. The high level of S induced only a limited decrease of total thiamin flow. Consequently, it is unlikely that the investigated factors could be considered to be high risk factors for the thiamin-dependent CCN. PMID:9389886

  18. Comparative Metabolite Fingerprinting of the Rumen System during Colonisation of Three Forage Grass (Lolium perenne L.) Varieties

    PubMed Central

    Kingston-Smith, Alison H.; Davies, Teri E.; Rees Stevens, Pauline; Mur, Luis A. J.

    2013-01-01

    The rumen microbiota enable ruminants to degrade complex ligno-cellulosic compounds to produce high quality protein for human consumption. However, enteric fermentation by domestic ruminants generates negative by-products: greenhouse gases (methane) and environmental nitrogen pollution. The current lack of cultured isolates representative of the totality of rumen microbial species creates an information gap about the in vivo function of the rumen microbiota and limits our ability to apply predictive biology for improvement of feed for ruminants. In this work we took a whole ecosystem approach to understanding how the metabolism of the microbial population responds to introduction of its substrate. Fourier Transform Infra Red (FTIR) spectroscopy-based metabolite fingerprinting was used to discriminate differences in the plant-microbial interactome of the rumen when using three forage grass varieties (Lolium perenne L. cv AberDart, AberMagic and Premium) as substrates for microbial colonisation and fermentation. Specific examination of spectral regions associated with fatty acids, amides, sugars and alkanes indicated that although the three forages were apparently similar by traditional nutritional analysis, patterns of metabolite flux within the plant-microbial interactome were distinct and plant genotype dependent. Thus, the utilisation pattern of forage nutrients by the rumen microbiota can be influenced by subtleties determined by forage genotypes. These data suggest that our interactomic approach represents an important means to improve forages and ultimately the livestock environment. PMID:24312434

  19. Microbial production of spider silk proteins.

    PubMed

    Fahnestock, S R; Yao, Z; Bedzyk, L A

    2000-08-01

    The remarkable properties of spider dragline silk and related protein polymers will find many applications if the materials can be produced economically. We have demonstrated the production of high molecular weight spider dragline silk analog proteins encoded by synthetic genes in several microbial systems, including Escherichia coli and Pichia pastoris. In E. coli, proteins of up to 1000 amino acids in length could be produced efficiently, but the yield and homogeneity of higher molecular weight silk proteins were found to be limited by truncated synthesis, probably as a result of ribosome termination errors. No such phenomenon was observed in the yeast P. pastoris, where higher molecular weight silk proteins could be produced without heterogeneity due to truncated synthesis. Spider dragline silk analog proteins could be secreted by P. pastoris when fused to both the signal sequence and N-terminal pro-sequence of the Saccharomyces cerevisiae alpha-mating factor gene. PMID:11763501

  20. Measurement of the Intestinal Digestibility of Rumen Undegraded Protein Using Different Methods and Correlation Analysis

    PubMed Central

    Wang, Y.; Zhang, Y. G.; Liu, Xiaolan; Kopparapu, N. K.; Xin, Hangshu; Liu, J.; Guo, Jianhua

    2015-01-01

    Four methods were adopted, including the mobile nylon bag (MNB) method, modified three-step in vitro (MTS) method, original three-step in vitro (OTS) method, and acid detergent insoluble nitrogen (ADIN) estimating method, to evaluate the intestinal digestibility of rumen undegradable protein (DRUP) of 10 types of concentrates and 7 types of roughages. After correlation analysis to determine the DRUP values using the MNB, MTS, OTS, and ADIN methods, the study aimed to find out appropriate methods to replace the MNB method due to its disadvantages such as high price, long time period, and use of a duodenal T-fistula. Three dairy cows with a permanent ruminal fistula and duodenal T-fistula were used in a single-factor experimental design. The results showed that the determined DRUP values using the MNB method for soybean meal, cottonseed meal, rapeseed meal, sunflower meal, corn germ meal, corn, rice bran, barley, wheat bran, corn fiber feed, Alfalfa (Zhao dong), Alfalfa (Long mu 801), Alfalfa (Long mu 803), grass (North), Grass (Inner Mongolia), corn silage and corn straw were 98.13%, 87.37%, 88.47%, 82.60%, 75.40%, 93.23%, 69.27%, 91.27%, 72.37%, 79.03%, 66.72%, 68.64%, 73.57%, 50.47%, 51.52%, 54.05%, and 43.84%, respectively. The coefficient of determination (R2 = 0.964) of the results between the MTS method and the MNB method was higher than that (R2 = 0.942) between the OTS method and the MNB method. The coefficient of determination of the DRUP values of the concentrates among the in vitro method (including the MTS and OTS methods) and the MNB method was higher than that of the roughage. There was a weak correlation between the determined DRUP values in concentrates obtained from the ADIN method and those from the MNB method, and there was a significant correlation (p<0.01) between the determined DRUP values of the roughage obtained from the MNB method and those obtained from ADIN method. The DRUP values were significantly correlated with the nutritional

  1. Insights into the bovine rumen plasmidome.

    PubMed

    Brown Kav, Aya; Sasson, Goor; Jami, Elie; Doron-Faigenboim, Adi; Benhar, Itai; Mizrahi, Itzhak

    2012-04-01

    Plasmids are self-replicating genetic elements capable of mobilization between different hosts. Plasmids often serve as mediators of lateral gene transfer, a process considered to be a strong and sculpting evolutionary force in microbial environments. Our aim was to characterize the overall plasmid population in the environment of the bovine rumen, which houses a complex and dense microbiota that holds enormous significance for humans. We developed a procedure for the isolation of total rumen plasmid DNA, termed rumen plasmidome, and subjected it to deep sequencing using the Illumina paired-end protocol and analysis using public and custom-made bioinformatics tools. A large number of plasmidome contigs aligned with plasmids of rumen bacteria isolated from different locations and at various time points, suggesting that not only the bacterial taxa, but also their plasmids, are defined by the ecological niche. The bacterial phylum distribution of the plasmidome was different from that of the rumen bacterial taxa. Nevertheless, both shared a dominance of the phyla Firmicutes, Bacteroidetes, and Proteobacteria. Evidently, the rumen plasmidome is of a highly mosaic nature that can cross phyla. Interestingly, when we compared the functional profile of the rumen plasmidome to two plasmid databases and two recently published rumen metagenomes, it became apparent that the rumen plasmidome codes for functions, which are enriched in the rumen ecological niche and could confer advantages to their hosts, suggesting that the functional profiles of mobile genetic elements are associated with their environment, as has been previously implied for viruses. PMID:22431592

  2. Metagenomic analysis of Surti buffalo (Bubalus bubalis) rumen: a preliminary study.

    PubMed

    Singh, Krishna M; Ahir, Viral B; Tripathi, Ajai K; Ramani, Umed V; Sajnani, Manisha; Koringa, Prakash G; Jakhesara, Subhash; Pandya, Paresh R; Rank, Dharamsi N; Murty, Duggirala S; Kothari, Ramesh K; Joshi, Chaitanya G

    2012-04-01

    The complex microbiome of the rumen functions as an effective system for the conversion of plant cell wall biomass to microbial proteins, short chain fatty acids and gases. In this study, metagenomic approaches were used to study the microbial populations and metabolic potential of the microbial community. DNA was extracted from Surti Buffalo rumen samples (four treatments diet) and sequenced separately using a 454 GS FLX Titanium system. We used comparative metagenomics to examine metabolic potential and phylogenetic composition from pyrosequence data generated in four samples, considering phylogenetic composition and metabolic potentials in the rumen may remarkably be different with respect to nutrient utilization. Assignment of metagenomic sequences to SEED categories of the Metagenome Rapid Annotation using Subsystem Technology (MG-RAST) server revealed a genetic profile characteristic of fermentation of carbohydrates in a high roughage diet. The distribution of phylotypes and environmental gene tags (EGTs) detected within each rumen sample were dominated by Bacteroidetes/Chlorobi, Firmicutes and Proteobacteria in all the samples. The results of this study could help to determine the role of rumen microbes and their enzymes in plant polysaccharide breakdown is fundamental to understanding digestion and maximising productivity in ruminant animals. PMID:21947953

  3. Metagenomic insights into the RDX-degrading potential of the ovine rumen microbiome.

    PubMed

    Li, Robert W; Giarrizzo, Juan Gabriel; Wu, Sitao; Li, Weizhong; Duringer, Jennifer M; Craig, A Morrie

    2014-01-01

    The manufacturing processes of royal demolition explosive (RDX), or hexahydro-1,3,5-trinitro-1,3,5-triazine, have resulted in serious water contamination. As a potential carcinogen, RDX can cause a broad range of harmful effects to humans and animals. The ovine rumen is capable of rapid degradation of nitroaromatic compounds, including RDX. While ruminal RDX-degrading bacteria have been identified, the genes and pathways responsible for RDX degradation in the rumen have yet to be characterized. In this study, we characterized the metabolic potential of the ovine rumen using metagenomic approaches. Sequences homologous to at least five RDX-degrading genes cloned from environmental samples (diaA, xenA, xenB, xplA, and xplB) were present in the ovine rumen microbiome. Among them, diaA was the most abundant, likely reflective of the predominance of the genus Clostridium in the ovine rumen. At least ten genera known to harbor RDX-degrading microorganisms were detectable. Metagenomic sequences were also annotated using public databases, such as Pfam, COG, and KEGG. Five of the six Pfam protein families known to be responsible for RDX degradation in environmental samples were identified in the ovine rumen. However, increased substrate availability did not appear to enhance the proliferation of RDX-degrading bacteria and alter the microbial composition of the ovine rumen. This implies that the RDX-degrading capacity of the ovine rumen microbiome is likely regulated at the transcription level. Our results provide metagenomic insights into the RDX-degrading potential of the ovine rumen, and they will facilitate the development of novel and economic bioremediation strategies.

  4. Metagenomic insights into the RDX-degrading potential of the ovine rumen microbiome.

    PubMed

    Li, Robert W; Giarrizzo, Juan Gabriel; Wu, Sitao; Li, Weizhong; Duringer, Jennifer M; Craig, A Morrie

    2014-01-01

    The manufacturing processes of royal demolition explosive (RDX), or hexahydro-1,3,5-trinitro-1,3,5-triazine, have resulted in serious water contamination. As a potential carcinogen, RDX can cause a broad range of harmful effects to humans and animals. The ovine rumen is capable of rapid degradation of nitroaromatic compounds, including RDX. While ruminal RDX-degrading bacteria have been identified, the genes and pathways responsible for RDX degradation in the rumen have yet to be characterized. In this study, we characterized the metabolic potential of the ovine rumen using metagenomic approaches. Sequences homologous to at least five RDX-degrading genes cloned from environmental samples (diaA, xenA, xenB, xplA, and xplB) were present in the ovine rumen microbiome. Among them, diaA was the most abundant, likely reflective of the predominance of the genus Clostridium in the ovine rumen. At least ten genera known to harbor RDX-degrading microorganisms were detectable. Metagenomic sequences were also annotated using public databases, such as Pfam, COG, and KEGG. Five of the six Pfam protein families known to be responsible for RDX degradation in environmental samples were identified in the ovine rumen. However, increased substrate availability did not appear to enhance the proliferation of RDX-degrading bacteria and alter the microbial composition of the ovine rumen. This implies that the RDX-degrading capacity of the ovine rumen microbiome is likely regulated at the transcription level. Our results provide metagenomic insights into the RDX-degrading potential of the ovine rumen, and they will facilitate the development of novel and economic bioremediation strategies. PMID:25383623

  5. Metagenomic Insights into the RDX-Degrading Potential of the Ovine Rumen Microbiome

    PubMed Central

    Li, Robert W.; Giarrizzo, Juan Gabriel; Wu, Sitao; Li, Weizhong; Duringer, Jennifer M.; Craig, A. Morrie

    2014-01-01

    The manufacturing processes of royal demolition explosive (RDX), or hexahydro-1,3,5-trinitro-1,3,5-triazine, have resulted in serious water contamination. As a potential carcinogen, RDX can cause a broad range of harmful effects to humans and animals. The ovine rumen is capable of rapid degradation of nitroaromatic compounds, including RDX. While ruminal RDX-degrading bacteria have been identified, the genes and pathways responsible for RDX degradation in the rumen have yet to be characterized. In this study, we characterized the metabolic potential of the ovine rumen using metagenomic approaches. Sequences homologous to at least five RDX-degrading genes cloned from environmental samples (diaA, xenA, xenB, xplA, and xplB) were present in the ovine rumen microbiome. Among them, diaA was the most abundant, likely reflective of the predominance of the genus Clostridium in the ovine rumen. At least ten genera known to harbor RDX-degrading microorganisms were detectable. Metagenomic sequences were also annotated using public databases, such as Pfam, COG, and KEGG. Five of the six Pfam protein families known to be responsible for RDX degradation in environmental samples were identified in the ovine rumen. However, increased substrate availability did not appear to enhance the proliferation of RDX-degrading bacteria and alter the microbial composition of the ovine rumen. This implies that the RDX-degrading capacity of the ovine rumen microbiome is likely regulated at the transcription level. Our results provide metagenomic insights into the RDX-degrading potential of the ovine rumen, and they will facilitate the development of novel and economic bioremediation strategies. PMID:25383623

  6. Manipulation of rumen ecology by dietary lemongrass (Cymbopogon citratus Stapf.) powder supplementation.

    PubMed

    Wanapat, M; Cherdthong, A; Pakdee, P; Wanapat, S

    2008-12-01

    This experiment was conducted to investigate the effect of lemongrass [Cymbopogon citratus (DC.) Stapf.] powder (LGP) on rumen ecology, rumen microorganisms, and digestibility of nutrients. Four ruminally fistulated crossbred (Brahman native) beef cattle were randomly assigned according to a 4 x 4 Latin square design. The dietary treatments were LGP supplementation at 0, 100, 200, and 300 g/d with urea-treated rice straw (5%) fed to allow ad libitum intake. Digestibilities of DM, ether extract, and NDF were significantly different among treatments and were greatest at 100 g/d of supplementation. However, digestibility of CP was decreased with LGP supplementation (P < 0.05), whereas ruminal NH(3)-N and plasma urea N were decreased with incremental additions of LGP (P < 0.05). Ruminal VFA concentrations were similar among supplementation concentrations (P > 0.05). Total viable bacteria, amylolytic bacteria, and cellulolytic bacteria were significantly different among treatments and were greatest at 100 g/d of supplementation (4.7 x 10(9), 1.7 x 10(7), and 2.0 x 10(9) cfu/mL, respectively). Protozoal populations were significantly decreased by LGP supplementation. In addition, efficiency of rumen microbial N synthesis based on OM truly digested in the rumen was enriched by LGP supplementation, especially at 100 g/d (34.2 g of N/kg of OM truly digested in the rumen). Based on this study, it could be concluded that supplementation of LGP at 100 g/d improved digestibilities of nutrients, rumen microbial population, and microbial protein synthesis efficiency, thus improving rumen ecology in beef cattle.

  7. Effect of donor animals and their diet on in vitro nutrient degradation and microbial protein synthesis using grass and corn silages.

    PubMed

    Boguhn, J; Zuber, T; Rodehutscord, M

    2013-06-01

    Two nonlactating cows and two wether sheep, all fitted with a permanent cannula into the rumen, were fed either hay plus concentrate, grass silage or corn silage to study the effect of the donor animal and its diet on in vitro fermentation and microbial protein synthesis. Rumen inoculum was obtained before the morning feeding. Grass silage or corn silage was incubated in a semi-continuous rumen simulation system for 14 days. Four replicated vessels were used per treatment. Degradation of crude nutrients and detergent fibre fractions as well as microbial protein synthesis and the production of volatile fatty acids were studied. Additionally, total gas and methane production was measured with a standard in vitro gas test. Gas production and methane concentration was higher when the inoculum used was from sheep than that from cows. The donor animal also affected the degradation of organic matter and ether extract as well as the amount of propionate and butyrate, and the acetate-to-propionate ratio. The effect of the diet fed to the donor animal on fermentation was much greater than the effect of the donor animal itself. Feeding hay plus concentrate resulted in higher gas production and degradation of acid detergent fibre, but in lower degradation of ether extract and reduced microbial protein synthesis. Additionally, the pattern of volatile fatty acids changed significantly when the diet of the donor animals was hay plus concentrate or one of the silages. These results show that in vitro fermentation and microbial protein synthesis is different when based on inoculum from either cattle or sheep. The diet fed to the donor animal is more important than the animal species and is probably mediated by an adjusted microbial activity. With regard to standardized feed evaluations, these results further support the need to harmonize in vitro approaches used in different laboratories.

  8. Changes in Microbial Diversity, Methanogenesis and Fermentation Characteristics in the Rumen in Response to Medicinal Plant Extracts

    PubMed Central

    Kim, Eun Tae; Moon, Yea Hwang; Min, Kwan-Sik; Kim, Chang-Hyun; Kim, Sam Churl; Ahn, Seung Kyu; Lee, Sung Sill

    2013-01-01

    This study evaluated the in vitro effect of medicinal plant extracts on ruminal methanogenesis, four different groups of methanogens and ruminal fermentation characteristics. A fistulated Holstein cow was used as a donor of rumen fluid. Licorice and mugwort extracts (Glycyrrhiza uralensis and Artemisia capillaris, 0.5% and 1% of total substrate DM, respectively), previously used as folk remedies, were added to an in vitro fermentation incubated with buffered-rumen fluid. Total gas production in Glycyrrhiza uralensis extract treatment was not significantly different between treatments (p<0.05) while total gas production in the Artemisia capillaris extract treatment was lower than that of the control. Artemisia capillaris extract and Glycyrrhiza uralensis extract reduced CH4 emission by 14% (p<0.05) and 8% (p<0.05), respectively. Ciliate-associated methanogens population decreased by 18% in the medicinal plant extracts treatments. Medicinal plant extracts also affected the order Methanobacteriales community. Methanobacteriales diversity decreased by 35% in the Glycyrrhiza uralensis extract treatment and 30% in the Artemisia capillaris extract treatment. The order Methanomicrobiales population decreased by 50% in the 0.5% of Glycyrrhiza uralensis extract treatment. These findings demonstrate that medicinal plant extracts have the potential to inhibit in vitro ruminal methanogenesis. PMID:25049911

  9. Brazilian propolis extract used as an additive to decrease methane emissions from the rumen microbial population in vitro.

    PubMed

    Santos, Nadine Woruby; Zeoula, Lucia Maria; Yoshimura, Emerson Henri; Machado, Erica; Macheboeuf, Didier; Cornu, Agnès

    2016-06-01

    Propolis is a product that is rich in phenolic compounds and can be utilized in animal nutrition as a dietary additive. In this study, the effects of a Brazilian green propolis extract on rumen fermentation and gas production were determined. The fate of propolis phenolic compounds in the rumen medium was also investigated. Fermentation was done in 24-h batches over three periods. Inoculates were obtained from cows fed on grassland hay and concentrate. Propolis extract in a hydroalcoholic solution was applied at increasing doses to the substrate (1 to 56 g/kg). The fermentation substrate consisted on a mixture of alfalfa hay, soybean meal, and wheat grain mixture in dry matter. After 24 h of fermentation, seven new compounds were observed in the medium in amounts that correlated to the propolis dose. The dose of propolis extract linearly decreased the pH of the medium and linearly increased propionate production, which reduced the acetate-to-propionate ratio and influenced the total production of short-chain fatty acids. Propolis also linearly reduced methane production and increased the carbon dioxide-to-methane ratio. Ammonia nitrogen levels and in vitro digestibility of organic matter were similar among the treatments. The combination of increased propionate production and decreased methane production suggests better energy utilization from the feed. PMID:27105625

  10. Pelleted beet pulp substituted for high-moisture corn: 3. Effects on ruminal fermentation, pH, and microbial protein efficiency in lactating dairy cows.

    PubMed

    Voelker, J A; Allen, M S

    2003-11-01

    The effects of increasing concentrations of dried, pelleted beet pulp substituted for high-moisture corn on ruminal fermentation, pH, and microbial efficiency were evaluated using eight ruminally and duodenally cannulated multiparous Holstein cows in a duplicated 4 x 4 Latin square design with 21-d periods. Cows were 79 +/- 17 (mean +/- SD) DIM at the beginning of the experiment. Experimental diets with 40% forage (corn silage and alfalfa silage) and 60% concentrate contained 0, 6.1, 12.1, or 24.3% beet pulp substituted for high-moisture corn on a DM basis. Diet concentrations of NDF and starch were 24.3 and 34.6% (0% beet pulp), 26.2 and 30.5% (6% beet pulp), 28.0, and 26.5% (12% beet pulp), and 31.6 and 18.4% (24% beet pulp), respectively. Substituting beet pulp for corn did not affect daily mean or minimum ruminal pH but tended to reduce pH range. Ruminal acetate:propionate responded in a positive exponential relationship to added beet pulp. Rate of valerate absorption from the rumen was not affected by treatment. Substituting beet pulp for corn up to 24% of diet DM did not affect efficiency of ruminal microbial protein production, expressed as microbial N flow to the duodenum as a percentage of OM truly digested in the rumen. Microbial efficiency was not correlated to mean pH or daily minimum pH. While microbial efficiency was not directly related to concentration of beet pulp fed, it was positively correlated with passage rate of particulate matter, as represented by starch and indigestible NDF, probably due to reduced turnover of microbial protein in the rumen.

  11. Effects of Synchronization of Carbohydrate and Protein Supply in Total Mixed Ration with Korean Rice Wine Residue on Ruminal Fermentation, Nitrogen Metabolism and Microbial Protein Synthesis in Holstein Steers

    PubMed Central

    Piao, Min Yu; Kim, Hyun J.; Seo, J. K.; Park, T. S.; Yoon, J. S.; Kim, K. H.; Ha, Jong K.

    2012-01-01

    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 3×3 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 NH3-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 NH3-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. PMID:25049518

  12. The use of a tannin crude extract from Cistus ladanifer L. to protect soya-bean protein from degradation in the rumen.

    PubMed

    Dentinho, M T P; Moreira, O C; Pereira, M S; Bessa, R J B

    2007-06-01

    Cistus ladanifer L. (CL) is a perennial shrub abundant in dry woods and dry land of Mediterranean zone, with high level of tannins. Tannins bind to protein, preventing its degradation in the digestive compartments. This tannin/protein complex may be advantageous when partially protecting good-quality feed protein from excessive rumen protein degradation. The objective of this trial was to use a CL phenol crude extract to prevent excessive rumen degradation of soya-bean meal protein. The phenolic compounds were extracted using an acetone/water solution (70:30, v/v). Soya-bean meal was then treated with this crude CL extract, containing 640 g of total phenols (TP) per kg of dry matter (DM), in order to obtain mixtures with 0, 12.5, 25, 50, 100 and 150 g of TP per kg DM. Three rumen-cannulated rams were used to assess in sacco rumen degradability of DM and nitrogen (N). The three-step in vitro procedure was used to determine intestinal digestibility. Increasing extract concentrations quadratically decreased the N-soluble fraction a (R2 = 0.96, P = 0.0001) and increased the non-soluble degradable fraction b (R2 = 0.92, P = 0.005). The rate of degradation c linearly decreased with CL extract doses (R2 = 0.44, P = 0.0065). For the effective rumen degradability of N, a linear reduction (R2 = 0.94, P < 0.0001) was observed. The in vitro intestinal digestibility of protein (ivID) quadratically decreased (R2 = 0.99, P < 0.0001) with TP inclusion and the rumen undegradable protein (RUP) showed a quadratic increase (R2 = 0.94, P = 0.0417). Total intestinal protein availability, computed from the RUP and ivID, linearly decreased with TP inclusion level (R2 = 0.45, P = 0.0033).

  13. Rumen Microbiology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ruminant animals such as cattle, sheep, and goats are able to digest low-quality fibrous feedstuffs because they maintain a mutually beneficial relationship with microorganisms resident in their forestomach, the rumen. Ruminal microorganisms are bacteria, protozoa, fungi, and viruses that live in a...

  14. Gene-centric metagenomics of the fiber-adherent bovine rumen microbiome reveals forage specific glycoside hydrolases

    PubMed Central

    Brulc, Jennifer M.; Antonopoulos, Dionysios A.; Berg Miller, Margret E.; Wilson, Melissa K.; Yannarell, Anthony C.; Dinsdale, Elizabeth A.; Edwards, Robert E.; Frank, Edward D.; Emerson, Joanne B.; Wacklin, Pirjo; Coutinho, Pedro M.; Henrissat, Bernard; Nelson, Karen E.; White, Bryan A.

    2009-01-01

    The complex microbiome of the rumen functions as an effective system for the conversion of plant cell wall biomass to microbial protein, short chain fatty acids, and gases. As such, it provides a unique genetic resource for plant cell wall degrading microbial enzymes that could be used in the production of biofuels. The rumen and gastrointestinal tract harbor a dense and complex microbiome. To gain a greater understanding of the ecology and metabolic potential of this microbiome, we used comparative metagenomics (phylotype analysis and SEED subsystems-based annotations) to examine randomly sampled pyrosequence data from 3 fiber-adherent microbiomes and 1 pooled liquid sample (a mixture of the liquid microbiome fractions from the same bovine rumens). Even though the 3 animals were fed the same diet, the community structure, predicted phylotype, and metabolic potentials in the rumen were markedly different with respect to nutrient utilization. A comparison of the glycoside hydrolase and cellulosome functional genes revealed that in the rumen microbiome, initial colonization of fiber appears to be by organisms possessing enzymes that attack the easily available side chains of complex plant polysaccharides and not the more recalcitrant main chains, especially cellulose. Furthermore, when compared with the termite hindgut microbiome, there are fundamental differences in the glycoside hydrolase content that appear to be diet driven for either the bovine rumen (forages and legumes) or the termite hindgut (wood). PMID:19181843

  15. Gene-centric metagenomics of the fiber-adherent bovine rumen microbiome reveals forage specific glycoside hydrolases.

    PubMed

    Brulc, Jennifer M; Antonopoulos, Dionysios A; Miller, Margret E Berg; Wilson, Melissa K; Yannarell, Anthony C; Dinsdale, Elizabeth A; Edwards, Robert E; Frank, Edward D; Emerson, Joanne B; Wacklin, Pirjo; Coutinho, Pedro M; Henrissat, Bernard; Nelson, Karen E; White, Bryan A

    2009-02-10

    The complex microbiome of the rumen functions as an effective system for the conversion of plant cell wall biomass to microbial protein, short chain fatty acids, and gases. As such, it provides a unique genetic resource for plant cell wall degrading microbial enzymes that could be used in the production of biofuels. The rumen and gastrointestinal tract harbor a dense and complex microbiome. To gain a greater understanding of the ecology and metabolic potential of this microbiome, we used comparative metagenomics (phylotype analysis and SEED subsystems-based annotations) to examine randomly sampled pyrosequence data from 3 fiber-adherent microbiomes and 1 pooled liquid sample (a mixture of the liquid microbiome fractions from the same bovine rumens). Even though the 3 animals were fed the same diet, the community structure, predicted phylotype, and metabolic potentials in the rumen were markedly different with respect to nutrient utilization. A comparison of the glycoside hydrolase and cellulosome functional genes revealed that in the rumen microbiome, initial colonization of fiber appears to be by organisms possessing enzymes that attack the easily available side chains of complex plant polysaccharides and not the more recalcitrant main chains, especially cellulose. Furthermore, when compared with the termite hindgut microbiome, there are fundamental differences in the glycoside hydrolase content that appear to be diet driven for either the bovine rumen (forages and legumes) or the termite hindgut (wood). PMID:19181843

  16. Gene-centric metagenomics of the fiber-adherent bovine rumen microbiome reveals forage specific glycoside hydrolases.

    PubMed

    Brulc, Jennifer M; Antonopoulos, Dionysios A; Miller, Margret E Berg; Wilson, Melissa K; Yannarell, Anthony C; Dinsdale, Elizabeth A; Edwards, Robert E; Frank, Edward D; Emerson, Joanne B; Wacklin, Pirjo; Coutinho, Pedro M; Henrissat, Bernard; Nelson, Karen E; White, Bryan A

    2009-02-10

    The complex microbiome of the rumen functions as an effective system for the conversion of plant cell wall biomass to microbial protein, short chain fatty acids, and gases. As such, it provides a unique genetic resource for plant cell wall degrading microbial enzymes that could be used in the production of biofuels. The rumen and gastrointestinal tract harbor a dense and complex microbiome. To gain a greater understanding of the ecology and metabolic potential of this microbiome, we used comparative metagenomics (phylotype analysis and SEED subsystems-based annotations) to examine randomly sampled pyrosequence data from 3 fiber-adherent microbiomes and 1 pooled liquid sample (a mixture of the liquid microbiome fractions from the same bovine rumens). Even though the 3 animals were fed the same diet, the community structure, predicted phylotype, and metabolic potentials in the rumen were markedly different with respect to nutrient utilization. A comparison of the glycoside hydrolase and cellulosome functional genes revealed that in the rumen microbiome, initial colonization of fiber appears to be by organisms possessing enzymes that attack the easily available side chains of complex plant polysaccharides and not the more recalcitrant main chains, especially cellulose. Furthermore, when compared with the termite hindgut microbiome, there are fundamental differences in the glycoside hydrolase content that appear to be diet driven for either the bovine rumen (forages and legumes) or the termite hindgut (wood).

  17. Comparison of utilization of pectins from various sources by pure cultures of pectinolytic rumen bacteria and mixed cultures of rumen microorganisms.

    PubMed

    Kasperowicz, A

    1994-01-01

    Utilization of citrus, lucerne, apple and sugar beet pulp pectins by pure strains of rumen bacteria, Prevotella ruminicola, Lachnospira multiparus and Butyrivibrio fibrisolvens was compared. Additionally, the utilization of pectins by mixed rumen microorganisms was evaluated. The comparison was based on the depletion of galacturonic acid from medium, content of cellular protein in the cultures and the amount of end products of pectin fermentation in cell-free culture fluids. It was found that citrus pectin was utilized best; utilization of lucerne, apple and sugar beet pectins was dependent on the species of bacteria. P. ruminicola and B. fibrisolvens utilized polygalacturonic acid from sugar beet pectins better than that from apple or lucerne pectin, while L. multiparus was capable of significantly better utilization of lucerne pectin than pectin from sugar beet or apple. The source of pectin was less important for mixed cultures of rumen microorganisms than for pure cultures of rumen bacteria. The amount of fermentation products in the culture fluids supported the conclusion that citrus pectin was utilized better than others. Microbial protein content in the cultures was found to be a less sensitive indicator of pectin utilization than the remaining examined parameters. P. ruminicola strains and mixed cultures of rumen microorganisms were shown to have the highest ability to utilize pectins, L. multiparus-moderate, while the B. fibrisolvens strains utilized pectin the least.

  18. Exploring the sheep rumen microbiome for carbohydrate-active enzymes.

    PubMed

    Lopes, Lucas Dantas; de Souza Lima, André Oliveira; Taketani, Rodrigo Gouvêa; Darias, Phillip; da Silva, Lília Raquel Fé; Romagnoli, Emiliana Manesco; Louvandini, Helder; Abdalla, Adibe Luiz; Mendes, Rodrigo

    2015-07-01

    The rumen is a complex ecosystem enriched for microorganisms able to degrade biomass during the animal's digestion process. The recovery of new enzymes from naturally evolved biomass-degrading microbial communities is a promising strategy to overcome the inefficient enzymatic plant destruction in industrial production of biofuels. In this context, this study aimed to describe the bacterial composition and functions in the sheep rumen microbiome, focusing on carbohydrate-active enzymes (CAE). Here, we used phylogenetic profiling analysis (inventory of 16S rRNA genes) combined with metagenomics to access the rumen microbiome of four sheep and explore its potential to identify fibrolytic enzymes. The bacterial community was dominated by Bacteroidetes and Firmicutes, followed by Proteobacteria. As observed for other ruminants, Prevotella was the dominant genus in the microbiome, comprising more than 30 % of the total bacterial community. Multivariate analysis of the phylogenetic profiling data and chemical parameters showed a positive correlation between the abundance of Prevotellaceae (Bacteroidetes phylum) and organic matter degradability. A negative correlation was observed between Succinivibrionaceae (Proteobacteria phylum) and methane production. An average of 2 % of the shotgun metagenomic reads was assigned to putative CAE when considering nine protein databases. In addition, assembled contigs allowed recognition of 67 putative partial CAE (NCBI-Refseq) representing 12 glycosyl hydrolase families (Pfam database). Overall, we identified a total of 28 lignocellulases, 22 amylases and 9 other putative CAE, showing the sheep rumen microbiome as a promising source of new fibrolytic enzymes.

  19. Exploring the sheep rumen microbiome for carbohydrate-active enzymes.

    PubMed

    Lopes, Lucas Dantas; de Souza Lima, André Oliveira; Taketani, Rodrigo Gouvêa; Darias, Phillip; da Silva, Lília Raquel Fé; Romagnoli, Emiliana Manesco; Louvandini, Helder; Abdalla, Adibe Luiz; Mendes, Rodrigo

    2015-07-01

    The rumen is a complex ecosystem enriched for microorganisms able to degrade biomass during the animal's digestion process. The recovery of new enzymes from naturally evolved biomass-degrading microbial communities is a promising strategy to overcome the inefficient enzymatic plant destruction in industrial production of biofuels. In this context, this study aimed to describe the bacterial composition and functions in the sheep rumen microbiome, focusing on carbohydrate-active enzymes (CAE). Here, we used phylogenetic profiling analysis (inventory of 16S rRNA genes) combined with metagenomics to access the rumen microbiome of four sheep and explore its potential to identify fibrolytic enzymes. The bacterial community was dominated by Bacteroidetes and Firmicutes, followed by Proteobacteria. As observed for other ruminants, Prevotella was the dominant genus in the microbiome, comprising more than 30 % of the total bacterial community. Multivariate analysis of the phylogenetic profiling data and chemical parameters showed a positive correlation between the abundance of Prevotellaceae (Bacteroidetes phylum) and organic matter degradability. A negative correlation was observed between Succinivibrionaceae (Proteobacteria phylum) and methane production. An average of 2 % of the shotgun metagenomic reads was assigned to putative CAE when considering nine protein databases. In addition, assembled contigs allowed recognition of 67 putative partial CAE (NCBI-Refseq) representing 12 glycosyl hydrolase families (Pfam database). Overall, we identified a total of 28 lignocellulases, 22 amylases and 9 other putative CAE, showing the sheep rumen microbiome as a promising source of new fibrolytic enzymes. PMID:25900454

  20. Persistence and functional impact of a microbial inoculant on native microbial community structure, nutrient digestion and fermentation characteristics in a rumen model.

    PubMed

    Ziemer, C J; Sharp, R; Stern, M D; Cotta, M A; Whitehead, T R; Stahl, D A

    2002-10-01

    Small sub-unit (SSU) rRNA-targeted oligonucleotide probes were used to monitor the persistence of a genetically engineered bacterium inoculated in model rumens. Eight dual flow continuous culture fermenters were operated with either standard artificial saliva buffer or buffer with chondroitin sulfate (0.5 g/l) added. After 168 h of operation, fermenters were inoculated with Bacteroides thetaiotaomicron BTX (BTX), at approximately 1% of total bacteria. B. thetaiotaomicron was quantified using a species-specific probe and shown to persist in fermenters 144 h after inoculation (relative abundance 0.48% and 1.42% of total SSU rRNA with standard and chondroitin sulfate buffers, respectively). No B. thetaiotaomicron SSU rRNA was detected in fermenter samples prior to inoculation with strain BTX. Relative abundances of Bacteria, Eucarya and Archaea were not affected by either inoculation or buffer type. Fiber digestion, in particular the hemicellulose fraction, increased after strain BTX addition. Chondroitin sulfate addition to the buffer increased bacterial nitrogen flow in fermenters, but did not alter fiber digestion. Neither inoculum nor buffer type altered total short chain fatty acid (VFA) concentrations but proportions of individual VFA differed. In model rumens, B. thetaiotaomicron BTX increased fiber digestion when added to mixed ruminal microbes, independent of chondroitin sulfate addition; but further study is needed to determine effects on other fiber-digesting bacteria.

  1. Reducing crude protein and rumen degradable protein with a constant concentration of rumen undegradable protein in the diet of dairy cows: Production performance, nutrient digestibility, nitrogen efficiency, and blood metabolites.

    PubMed

    Bahrami-Yekdangi, M; Ghorbani, G R; Khorvash, M; Khan, M A; Ghaffari, M H

    2016-02-01

    The goals of ruminant protein nutrition are to provide adequate amounts of RDP for optimal ruminal efficiency and to obtain the desired animal productivity with a minimum amount of dietary CP. The aim of the present study was to examine effects of decreasing dietary protein by decreasing RDP with the optimum concentration of RUP on production performance, nutrient digestibility, N retention, rumen fermentation parameters, and blood metabolites in high-producing Holstein cows in early lactation. Nine multiparous lactating cows (second parities, averaging 50 ± 12 d in milk and milk yield of 48 ± 5 kg/d) were used in a triplicate 3 × 3 Latin square design with 3 rations: 1) a total mixed ration (TMR) containing 16.4% CP (10.9% RDP based on DM), 2) a TMR containing 15.6% CP (10% RDP), and 3) a TMR containing 14.8% CP (9.3% RDP). The level of RUP was constant at 5.5% DM across the treatments. All diets were calculated to supply a postruminal lysine to methionine ratio of about 3:1. Dry matter intake, milk yield and composition, 4% fat-corrected milk, and energy-corrected milk were not significantly affected by decreasing dietary CP and RDP levels. Cows fed 16.4% CP diets had greater ( < 0.01) CP and RDP intakes, which resulted in a trend toward greater concentrations of plasma urea N compared with other treatments. Daily N intake linearly decreased ( < 0.01) with decreasing dietary CP and RDP levels, whereas the intake of RUP and fecal N excretion (g/d) did not change. Apparent digestibility of nutrients, ruminal pH, and NH-N concentration were not affected with decreasing dietary CP and RDP levels. Apparent N efficiency increased, and RDP N intake and predicted urine N output decreased with decreased concentration of dietary CP and RDP in the diets ( < 0.01). Blood metabolites were not affected by treatments. In conclusion, to improve the efficiency of N utilization by early-lactation dairy cows, 9.3% RDP in rations provides adequate protein to optimize milk

  2. Protein Secondary Structures (alpha-helix and beta-sheet) at a Cellular Levle and Protein Fractions in Relation to Rumen Degradation Behaviours of Protein: A New Approach

    SciTech Connect

    Yu,P.

    2007-01-01

    Studying the secondary structure of proteins leads to an understanding of the components that make up a whole protein, and such an understanding of the structure of the whole protein is often vital to understanding its digestive behaviour and nutritive value in animals. The main protein secondary structures are the {alpha}-helix and {beta}-sheet. The percentage of these two structures in protein secondary structures influences protein nutritive value, quality and digestive behaviour. A high percentage of {beta}-sheet structure may partly cause a low access to gastrointestinal digestive enzymes, which results in a low protein value. The objectives of the present study were to use advanced synchrotron-based Fourier transform IR (S-FTIR) microspectroscopy as a new approach to reveal the molecular chemistry of the protein secondary structures of feed tissues affected by heat-processing within intact tissue at a cellular level, and to quantify protein secondary structures using multicomponent peak modelling Gaussian and Lorentzian methods, in relation to protein digestive behaviours and nutritive value in the rumen, which was determined using the Cornell Net Carbohydrate Protein System. The synchrotron-based molecular chemistry research experiment was performed at the National Synchrotron Light Source at Brookhaven National Laboratory, US Department of Energy. The results showed that, with S-FTIR microspectroscopy, the molecular chemistry, ultrastructural chemical make-up and nutritive characteristics could be revealed at a high ultraspatial resolution ({approx}10 {mu}m). S-FTIR microspectroscopy revealed that the secondary structure of protein differed between raw and roasted golden flaxseeds in terms of the percentages and ratio of {alpha}-helixes and {beta}-sheets in the mid-IR range at the cellular level. By using multicomponent peak modelling, the results show that the roasting reduced (P <0.05) the percentage of {alpha}-helixes (from 47.1% to 36.1%: S

  3. Effects of calcium soaps and rumen undegradable protein on the milk production and composition of dairy ewes.

    PubMed

    Casals, R; Caja, G; Such, X; Torre, C; Calsamiglia, S

    1999-05-01

    Forty-eight Manchega dairy ewes were used during a complete lactation in a 2 x 2 factorial design to determine the effects of supplementing diets with fat (calcium soaps of palm oil fatty acids, CSFA) and rumen undegradable protein (RUP) on milk production and composition. Factors tested were amounts of CSFA (0 or 200 g/kg) and RUP (300 or 450 g/kg crude protein) in the concentrate. RUP was altered by adding a mixture of maize gluten meal and blood meal. Lactation was divided into one nursing period (period 1, weeks 1-4), and three milking periods (periods 2-4, weeks 5-8, 9-14 and 15-21). Concentrates were given at 0.8 kg/d during periods 1 and 2, and at 0.6 kg/d in periods 3 and 4. Ewes grazed rotationally in an Italian rye-grass pasture and received a daily supplement of 0.8 kg vetch-oat hay during period 1, and 0.3 kg lucerne hay during periods 2-4. For the whole lactation, supplemental fat markedly increased milk fat content (+23%) and yield (+16%), and decreased milk protein content (-9%). The positive effect of feeding CSFA on milk fat content was more evident at the beginning of lactation; however, its negative effect on milk protein was more pronounced in late lactation. Supplementary RUP had little effect, increasing milk protein content only in period 3, when the crude protein content of pasture was lower. Milk yield and lamb growth were not affected by dietary treatments. The results indicated that CSFA can be useful for increasing the milk fat content of dairy ewes at pasture, which may help farmers to produce milk reaching the minimum requirements of fat content for the cheese industry.

  4. Microbial protein production: maximizing protein production efficiency in Space habitats

    NASA Astrophysics Data System (ADS)

    Clauwaert, Peter; Alloul, Abbas; Muys, Maarten; Sui, Yixing; Boon, Nico; Luther, Amanda; Christiaens, Marlies E. R.; Ilgrande, Chiara; Lindeboom, Ralph E. F.; Rabaey, Korneel; Vlaeminck, Siegfried

    2016-07-01

    On top of the goal of a closed material cycle for Space habitats or deep Space missions with food production, extreme requirements apply to such Life Support Systems (LSS) in terms of mass, volume, crew time, energy consumption and controllability. Although relatively high water recovery efficiencies (~70-90%) can be achieved, all Space missions until now have relied on terrestrial food resupply and thus no nutrient recovery has been achieved so far. Researchers and Space agencies have typically been focussing on the cultivation of higher plants to produce food for crew members for future Space LSS. It can be assumed that the required surface area (50-500 m2 per crew member), plant evaporation rates (~200 kg per crew member per day), power consumption (~65 kW per crew member) and the degree of controllability of a higher plant compartment will have a great impact on the feasibility of realizing a future closed loop LSS in Space for the first time. As the food production density is so critical in a LSS, a combination of higher plant cultivation and microbial protein production might increase the chances of success of future Space LSS's since the production densities are significantly higher. Higher plants in Space LSS's would typically have an average specific protein production rate in the order of 0-4 kg protein m-3 year-1 (calculated from Do, Owens et al. (2016)), whereas bacterial biomass can be produced continuously at a rate up to ~1000 kg protein m-3 year-1. Several routes for microbial food production will be discussed in this presentation, ranging from aerobic heterotrophic production with for instance Candida ingens (Strayer, Finger et al. 1997), photoheterotrophic production with PNSB such as Rhodospirillum rubrum (Hendrickx, De Wever et al. 2006) and hydrogenotrophic production with HOB such as Cupriavidus necator (Matassa, Boon et al. 2015)) and photoautotrophic production of oxygen and microbial food (e.g. Arthrospira sp. (Hendrickx, De Wever et al

  5. Metagenomic Insights into the Fibrolytic Microbiome in Yak Rumen

    PubMed Central

    Song, Lei; Liu, Di; Liu, Li; Chen, Furong; Wang, Min; Li, Jiabao; Zeng, Xiaowei; Dong, Zhiyang; Hu, Songnian; Li, Lingyan; Xu, Jian; Huang, Li; Dong, Xiuzhu

    2012-01-01

    The rumen hosts one of the most efficient microbial systems for degrading plant cell walls, yet the predominant cellulolytic proteins and fibrolytic mechanism(s) remain elusive. Here we investigated the cellulolytic microbiome of the yak rumen by using a combination of metagenome-based and bacterial artificial chromosome (BAC)-based functional screening approaches. Totally 223 fibrolytic BAC clones were pyrosequenced and 10,070 ORFs were identified. Among them 150 were annotated as the glycoside hydrolase (GH) genes for fibrolytic proteins, and the majority (69%) of them were clustered or linked with genes encoding related functions. Among the 35 fibrolytic contigs of >10 Kb in length, 25 were derived from Bacteroidetes and four from Firmicutes. Coverage analysis indicated that the fibrolytic genes on most Bacteroidetes-contigs were abundantly represented in the metagenomic sequences, and they were frequently linked with genes encoding SusC/SusD-type outer-membrane proteins. GH5, GH9, and GH10 cellulase/hemicellulase genes were predominant, but no GH48 exocellulase gene was found. Most (85%) of the cellulase and hemicellulase proteins possessed a signal peptide; only a few carried carbohydrate-binding modules, and no cellulosomal domains were detected. These findings suggest that the SucC/SucD-involving mechanism, instead of one based on cellulosomes or the free-enzyme system, serves a major role in lignocellulose degradation in yak rumen. Genes encoding an endoglucanase of a novel GH5 subfamily occurred frequently in the metagenome, and the recombinant proteins encoded by the genes displayed moderate Avicelase in addition to endoglucanase activities, suggesting their important contribution to lignocellulose degradation in the exocellulase-scarce rumen. PMID:22808161

  6. Metagenomic insights into the fibrolytic microbiome in yak rumen.

    PubMed

    Dai, Xin; Zhu, Yaxin; Luo, Yingfeng; Song, Lei; Liu, Di; Liu, Li; Chen, Furong; Wang, Min; Li, Jiabao; Zeng, Xiaowei; Dong, Zhiyang; Hu, Songnian; Li, Lingyan; Xu, Jian; Huang, Li; Dong, Xiuzhu

    2012-01-01

    The rumen hosts one of the most efficient microbial systems for degrading plant cell walls, yet the predominant cellulolytic proteins and fibrolytic mechanism(s) remain elusive. Here we investigated the cellulolytic microbiome of the yak rumen by using a combination of metagenome-based and bacterial artificial chromosome (BAC)-based functional screening approaches. Totally 223 fibrolytic BAC clones were pyrosequenced and 10,070 ORFs were identified. Among them 150 were annotated as the glycoside hydrolase (GH) genes for fibrolytic proteins, and the majority (69%) of them were clustered or linked with genes encoding related functions. Among the 35 fibrolytic contigs of >10 Kb in length, 25 were derived from Bacteroidetes and four from Firmicutes. Coverage analysis indicated that the fibrolytic genes on most Bacteroidetes-contigs were abundantly represented in the metagenomic sequences, and they were frequently linked with genes encoding SusC/SusD-type outer-membrane proteins. GH5, GH9, and GH10 cellulase/hemicellulase genes were predominant, but no GH48 exocellulase gene was found. Most (85%) of the cellulase and hemicellulase proteins possessed a signal peptide; only a few carried carbohydrate-binding modules, and no cellulosomal domains were detected. These findings suggest that the SucC/SucD-involving mechanism, instead of one based on cellulosomes or the free-enzyme system, serves a major role in lignocellulose degradation in yak rumen. Genes encoding an endoglucanase of a novel GH5 subfamily occurred frequently in the metagenome, and the recombinant proteins encoded by the genes displayed moderate Avicelase in addition to endoglucanase activities, suggesting their important contribution to lignocellulose degradation in the exocellulase-scarce rumen. PMID:22808161

  7. Effect of progressive inoculation of fauna-free sheep with holotrich protozoa and total-fauna on rumen fermentation, microbial diversity and methane emissions.

    PubMed

    Belanche, Alejandro; de la Fuente, Gabriel; Newbold, Charles J

    2015-03-01

    Rumen methanogenesis represents an energy waste for the ruminant and an important source of greenhouse gas; thus, integrated studies are needed to fully understand this process. Eight fauna-free sheep were used to investigate the effect of successive inoculation with holotrich protozoa then with total fauna on rumen methanogenesis. Holotrichs inoculation neither altered rumen fermentation rate nor diet digestibility, but increased concentrations of acetate (+15%), butyrate (+57%), anaerobic fungi (+0.82 log), methanogens (+0.41 log) and methanogenesis (+54%). Further inoculation with total fauna increased rumen concentrations of protozoa (+1.0 log), bacteria (+0.29 log), anaerobic fungi (+0.78 log), VFA (+8%), ammonia and fibre digestibility (+17%) without affecting levels of methanogens or methanogenesis. Rumen methanogens population was fairly stable in terms of structure and diversity, while the bacterial community was highly affected by the treatments. Inoculation with holotrich protozoa increased bacterial diversity. Further inoculation with total fauna lowered bacterial diversity but increased concentrations of certain propionate and lactate-producing bacteria, suggesting that alternative H2 sinks could be relevant. This experiment suggests that holotrich protozoa have a greater impact on rumen methanogenesis than entodiniomorphids. Thus, further research is warranted to understand the effect of holotrich protozoa on methane formation and evaluate their elimination from the rumen as a potential methane mitigation strategy.

  8. Characterization of the rumen microbiome of Indian Kankrej cattle (Bos indicus) adapted to different forage diet.

    PubMed

    Patel, Vilas; Patel, Amrutlal K; Parmar, Nidhi R; Patel, Anand B; Reddy, Bhaskar; Joshi, Chaitanya G

    2014-12-01

    Present study described rumen microbiome of Indian cattle (Kankrej breed) to better understand the microbial diversity and largely unknown functional capacity of the rumen microbiome under different dietary treatments. Kankrej cattle were gradually adapted to a high-forage diet (four animals with dry forage and four with green forage) containing 50 % (K1), 75 % (K2) to 100 % (K3) forage, and remaining concentrate diet, each for 6 weeks followed by analysis of rumen fiber adherent and fiber-free metagenomic community by shotgun sequencing using ion torrent PGM platform and EBI-metagenomics annotation pipeline. Taxonomic analysis indicated that rumen microbiome was dominated by Bacteroidetes followed by Firmicutes, Fibrobacter, Proteobacteria, and Tenericutes. Functional analysis based on gene ontology classified all reads in total 157 categories based on their functional role in biological, molecular, and cellular component with abundance of genes associated with hydrolase activity, membrane, transport, transferase, and different metabolism (such as carbohydrate and protein). Statistical analysis using STAMP revealed significant differences (P < 0.05) between solid and liquid fraction of rumen (in 65 categories), between all three treatments (in 56 categories), and between green and dry roughage (17 categories). Diet treatment also exerted significant difference in environmental gene tags (EGTs) involved in metabolic pathways for production of volatile fatty acids. EGTs for butyrate production were abundant in K2, whereas EGTs for propionate production was abundant during K1. Principal component analysis also demonstrated that diet proportion, fraction of rumen, and type of forage affected rumen microbiome at taxonomic as well as functional level. PMID:25359471

  9. Characterization of the rumen microbiome of Indian Kankrej cattle (Bos indicus) adapted to different forage diet.

    PubMed

    Patel, Vilas; Patel, Amrutlal K; Parmar, Nidhi R; Patel, Anand B; Reddy, Bhaskar; Joshi, Chaitanya G

    2014-12-01

    Present study described rumen microbiome of Indian cattle (Kankrej breed) to better understand the microbial diversity and largely unknown functional capacity of the rumen microbiome under different dietary treatments. Kankrej cattle were gradually adapted to a high-forage diet (four animals with dry forage and four with green forage) containing 50 % (K1), 75 % (K2) to 100 % (K3) forage, and remaining concentrate diet, each for 6 weeks followed by analysis of rumen fiber adherent and fiber-free metagenomic community by shotgun sequencing using ion torrent PGM platform and EBI-metagenomics annotation pipeline. Taxonomic analysis indicated that rumen microbiome was dominated by Bacteroidetes followed by Firmicutes, Fibrobacter, Proteobacteria, and Tenericutes. Functional analysis based on gene ontology classified all reads in total 157 categories based on their functional role in biological, molecular, and cellular component with abundance of genes associated with hydrolase activity, membrane, transport, transferase, and different metabolism (such as carbohydrate and protein). Statistical analysis using STAMP revealed significant differences (P < 0.05) between solid and liquid fraction of rumen (in 65 categories), between all three treatments (in 56 categories), and between green and dry roughage (17 categories). Diet treatment also exerted significant difference in environmental gene tags (EGTs) involved in metabolic pathways for production of volatile fatty acids. EGTs for butyrate production were abundant in K2, whereas EGTs for propionate production was abundant during K1. Principal component analysis also demonstrated that diet proportion, fraction of rumen, and type of forage affected rumen microbiome at taxonomic as well as functional level.

  10. Soybean meal substitution with a yeast-derived microbial protein source in dairy cow diets.

    PubMed

    Sabbia, J A; Kalscheur, K F; Garcia, A D; Gehman, A M; Tricarico, J M

    2012-10-01

    The objective of this study was to examine the effects substituting soybean meal with a yeast-derived microbial protein (YMP) on rumen and blood metabolites, dry matter intake, and milk production of high-producing dairy cows. Sixteen Holstein cows (12 multiparous and 4 primiparous), 93 ± 37 DIM (mean ± SD) at the beginning of the experiment, were used in a 4×4 Latin square design with four 28-d periods. Cows were blocked by parity and production, with 1 square consisting of 4 animals fitted with rumen cannulas. Basal diets, formulated for 16.1% crude protein and 1.56 Mcal/kg of net energy for lactation, contained 40% corn silage, 20% alfalfa hay, and 40% concentrate mix. During each period, cows were fed 1 of 4 treatment diets corresponding to YMP (DEMP; Alltech Inc., Nicholasville, KY) concentrations of 0, 1.14, 2.28, and 3.41% DM. Soybean meal (44% CP) was replaced by YMP to attain isonitrogenous and isoenergetic diets. Dietary treatments had no effect on pH and on most ruminal volatile fatty acid concentrations, with the exception of isovalerate, which decreased linearly with the addition of YMP. Rumen ammonia concentration decreased linearly, whereas free amino acids, total amino acid nitrogen, and soluble proteins weighing more than 10 kDa showed a cubic response on rumen N fractionation. A quadratic response was observed in oligopeptides that weighed between 3 and 10 kDa and peptides under 3kDa when expressed as percentages of total amino acids and total nitrogen. Although nonesterified fatty acid concentration in blood did not differ between treatments, β-hydroxybutyrate and plasma glucose increased linearly as YMP increased. Dry matter intake showed a cubic effect, where cows fed 1.14, and 3.41% YMP had the highest intake. Milk production was not affected by YMP, whereas a trend was observed for a quadratic increase for 4% fat-corrected milk and energy-corrected milk. Medium- and long-chain fatty acid concentrations in milk increased quadratically

  11. The Escherichia coli O157:H7 bovine rumen fluid proteome reflects adaptive bacterial responses

    PubMed Central

    2014-01-01

    Background To obtain insights into Escherichia coli O157:H7 (O157) survival mechanisms in the bovine rumen, we defined the growth characteristics and proteome of O157 cultured in rumen fluid (RF; pH 6.0-7.2 and low volatile fatty acid content) obtained from rumen-fistulated cattle fed low protein content “maintenance diet” under diverse in vitro conditions. Results Bottom-up proteomics (LC-MS/MS) of whole cell-lysates of O157 cultured under anaerobic conditions in filter-sterilized RF (fRF; devoid of normal ruminal microbiota) and nutrient-depleted and filtered RF (dRF) resulted in an anaerobic O157 fRF-and dRF-proteome comprising 35 proteins functionally associated with cell structure, motility, transport, metabolism and regulation, but interestingly, not with O157 virulence. Shotgun proteomics-based analysis using isobaric tags for relative and absolute quantitation used to further study differential protein expression in unfiltered RF (uRF; RF containing normal rumen microbial flora) complemented these results. Conclusions Our results indicate that in the rumen, the first anatomical compartment encountered by this human pathogen within the cattle gastrointestinal tract (GIT), O157 initiates a program of specific gene expression that enables it to adapt to the in vivo environment, and successfully transit to its colonization sites in the bovine GIT. Further experiments in vitro using uRF from animals fed different diets and with additional O157 strains, and in vivo using rumen-fistulated cattle will provide a comprehensive understanding of the adaptive mechanisms involved, and help direct evolution of novel modalities for blocking O157 infection of cattle. PMID:24559513

  12. [The influence of oral niacin doses during different dietary protein levels on indexes of rumen fermentation, blood parameters and fattening performance of young bulls].

    PubMed

    Flachowsky, G; Wolfram, D; Wilk, H; Schneider, M

    1993-01-01

    The influence of added niacin (0, 0.5 or 1 g per animal per day) and different crude protein contents (9.2 to 12.0% of DM) or the supply of various N-sources (urea, rape seed meal, soya bean meal, fish meal) were investigated on rumen fermentation, blood parameters, feed intake, weight gain and dry matter (DM) intake per weight gain in three individual feeding experiments with 156 growing bulls weighing between 175 and 300 kg per animal. Niacin supplementation did not significantly influence (P > 0.05) investigated parameters of rumen fermentation and blood. Ruminal propionate concentration increased insignificantly (from 18.9 to 19.5 moles per 100 moles on the average), inorganic P of blood serum somewhat decreased (from 2.98 to 2.82 mmol per 1) when niacin was added (P > 0.05). Protein level did not significantly influence rumen fermentation and blood parameters except an increased urea concentration in the blood of cattle fed with diets richer in protein. The DM intake of control bulls and niacin supplemented animals amounted to 6.35 and 6.46 kg per animal per day on average. Influence of niacin on DM intake varied in dependence on protein source. The daily weight gain increased from 1003 (control) to 1040 g per animal per day (+ niacin, P > 0.05). Niacin increased weight gain of bulls of urea (+ 43 g) and rape seed or soya bean meal added rations (+ 60 g per animal per day), but did not influence the weight gain in fish meal added rations. Increased weight gain resulted from ruminal and metabolic effects (about 2/3) as well as enhanced feed intake of bulls (about 1/3). Feed efficiency mostly improved. PMID:7487473

  13. Phage-bacteria relationships and CRISPR elements revealed by a metagenomic survey of the rumen microbiome.

    PubMed

    Berg Miller, Margret E; Yeoman, Carl J; Chia, Nicholas; Tringe, Susannah G; Angly, Florent E; Edwards, Robert A; Flint, Harry J; Lamed, Raphael; Bayer, Edward A; White, Bryan A

    2012-01-01

    Viruses are the most abundant biological entities on the planet and play an important role in balancing microbes within an ecosystem and facilitating horizontal gene transfer. Although bacteriophages are abundant in rumen environments, little is known about the types of viruses present or their interaction with the rumen microbiome. We undertook random pyrosequencing of virus-enriched metagenomes (viromes) isolated from bovine rumen fluid and analysed the resulting data using comparative metagenomics. A high level of diversity was observed with up to 28,000 different viral genotypes obtained from each environment. The majority (~78%) of sequences did not match any previously described virus. Prophages outnumbered lytic phages approximately 2:1 with the most abundant bacteriophage and prophage types being associated with members of the dominant rumen phyla (Firmicutes and Proteobacteria). Metabolic profiling based on SEED subsystems revealed an enrichment of sequences with putative functional roles in DNA and protein metabolism, but a surprisingly low proportion of sequences assigned to carbohydrate and amino acid metabolism. We expanded our analysis to include previously described metagenomic data and 14 reference genomes. Clustered regularly interspaced short palindromic repeats (CRISPR) were detected in most of the microbial genomes, suggesting previous interactions between viral and microbial communities.

  14. Phage-bacteria relationships and CRISPR elements revealed by a metagenomic survey of the rumen microbiome.

    PubMed

    Berg Miller, Margret E; Yeoman, Carl J; Chia, Nicholas; Tringe, Susannah G; Angly, Florent E; Edwards, Robert A; Flint, Harry J; Lamed, Raphael; Bayer, Edward A; White, Bryan A

    2012-01-01

    Viruses are the most abundant biological entities on the planet and play an important role in balancing microbes within an ecosystem and facilitating horizontal gene transfer. Although bacteriophages are abundant in rumen environments, little is known about the types of viruses present or their interaction with the rumen microbiome. We undertook random pyrosequencing of virus-enriched metagenomes (viromes) isolated from bovine rumen fluid and analysed the resulting data using comparative metagenomics. A high level of diversity was observed with up to 28,000 different viral genotypes obtained from each environment. The majority (~78%) of sequences did not match any previously described virus. Prophages outnumbered lytic phages approximately 2:1 with the most abundant bacteriophage and prophage types being associated with members of the dominant rumen phyla (Firmicutes and Proteobacteria). Metabolic profiling based on SEED subsystems revealed an enrichment of sequences with putative functional roles in DNA and protein metabolism, but a surprisingly low proportion of sequences assigned to carbohydrate and amino acid metabolism. We expanded our analysis to include previously described metagenomic data and 14 reference genomes. Clustered regularly interspaced short palindromic repeats (CRISPR) were detected in most of the microbial genomes, suggesting previous interactions between viral and microbial communities. PMID:22004549

  15. Effects of vanillin, quillaja saponin, and essential oils on in vitro fermentation and protein-degrading microorganisms of the rumen.

    PubMed

    Patra, Amlan K; Yu, Zhongtang

    2014-01-01

    This study investigated the effects of vanillin on methanogenesis and rumen fermentation, and the responses of ruminal protein-degrading bacteria to vanillin (at concentrations of 0, 0.76 and 1.52 g/L), essential oils (clove oil, 1 g/L; origanum oil, 0.50 g/L, and peppermint oil, 1 g/L), and quillaja saponin (at concentration of 0 and 6 g/L) in vitro. Methane production, degradabilities of feed substrate, and ammonia concentration decreased linearly with increasing doses of vanillin. Concentration of total volatile fatty acids also decreased, whereas proportion of butyrate tended to increase linearly with increasing doses of vanillin. Protozoa population decreased, but abundances of Ruminococcus flavefaciens, Prevotella bryantii, Butyrivibrio fibrisolvens, Prevotella ruminicola, Clostridium aminophilum, and Ruminobacter amylophilus increased with increasing doses of vanillin. Origanum and clove oils resulted in lower ammonia concentrations compared to control and peppermint oil. All the tested essential oils decreased abundances of protozoa, Selenomonas ruminantium, R. amylophilus, P. ruminicola and P. bryantii, with the largest decrease resulted from origanum oil followed by clove oil and peppermint oil. The abundances of Megasphaera elsdenii, C. aminophilum, and Clostridium sticklandii were deceased by origanum oil while that of B. fibrisolvens was lowered by both origanum and clove oils. Saponin decreased ammonia concentration and protozoal population, but increased the abundances of S. ruminantium, R. amylophilus, P. ruminicola, and P. bryantii, though the magnitude was small (less than one log unit). The results suggest that reduction of ammonia production by vanillin and saponin may not be caused by direct inhibition of major known proteolytic bacteria, and essential oils can have different inhibitory effects on different proteolytic bacteria, resulting in varying reduction in ammonia production.

  16. Application of rumen microorganisms for anaerobic bioconversion of lignocellulosic biomass.

    PubMed

    Yue, Zheng-Bo; Li, Wen-Wei; Yu, Han-Qing

    2013-01-01

    Rumen in the mammalian animals is a natural cellulose-degrading system and the microorganisms inside have been found to be able to effectively digest lignocellulosic biomass. Furthermore, methane or volatile fatty acids, which could be further converted to other biofuels, are the two major products in such a system. This paper offers an overview of recent development in the application of rumen microorganisms for lignocellulosic biomass conversion. Application of recent molecular tools in the analysis of rumen microbial community, progress in the development of artificial rumen reactors, the latest research results about characterizing rumen-dominated anaerobic digestion process and energy products are summarized. Also, the potential application of such a rumen-dominated process is discussed.

  17. The requirements for rumen-degradable protein per unit of fermentable organic matter differ between fibrous feed sources.

    PubMed

    Soliva, Carla R; Amelchanka, Sergej L; Kreuzer, Michael

    2015-01-01

    Ruminant feed evaluation systems use constant minimum requirements of rumen-degradable protein (RDP) and often relate this to apparently degradable organic matter (OM). However, studies with tropical forages indicate that RDP: apparently degraded OM might not be constant across high-fiber diets. This was tested with semi-continuous ruminal cultures (Rusitec) using dried contrasting low-protein fiber sources: brachiaria hay (high in fiber, medium lignified), apple pomace (medium in fiber, highly lignified), and sugar beet pulp (medium in fiber and lignification). Each feed was incubated at 14 g dry matter day(-1) with 0, 0.85, 1.7, 3.4, 6.8, 13.6, or 27.2 mg g(-1) urea. The amount of urea needed to reach a similar basal concentration of ammonia in the incubation fluid was tested for each feed in advance. Apparent fiber and OM degradability were determined after 48 h of incubation. Data was evaluated by regressions and analysis of variance. The response curve of incubation fluid ammonia to urea supplementation was similar in slope in all feeds. Plateaus in apparent OM degradability in relation to ammonia concentration were determined. The ammonia concentration where apparent OM and fiber degradability reached 95% of maximum was approached in the order of pomace < pulp < hay. With regard to fiber degradability, a plateau was reached at ≥ 80 g kg(-1) crude protein only with hay and pomace, whilst a linear relationship existed between RDP and OM degradation for pulp. In hay the ratio RDP: OM degraded was equal to 1.6 but was only 1.0 in the other feeds. There was no obvious lack of branched short-chain fatty acids at low RDP. Thus, the hypothesis was confirmed but the demand for RDP seems even higher in tropical forage compared to food industrial byproducts. The efficiency of urea to promote apparent OM and fiber degradation was also variable. Thus, it seems that minimum thresholds of either RDP or ruminal ammonia concentration may not be reflected appropriately by

  18. The requirements for rumen-degradable protein per unit of fermentable organic matter differ between fibrous feed sources

    PubMed Central

    Soliva, Carla R.; Amelchanka, Sergej L.; Kreuzer, Michael

    2015-01-01

    Ruminant feed evaluation systems use constant minimum requirements of rumen-degradable protein (RDP) and often relate this to apparently degradable organic matter (OM). However, studies with tropical forages indicate that RDP: apparently degraded OM might not be constant across high-fiber diets. This was tested with semi-continuous ruminal cultures (Rusitec) using dried contrasting low-protein fiber sources: brachiaria hay (high in fiber, medium lignified), apple pomace (medium in fiber, highly lignified), and sugar beet pulp (medium in fiber and lignification). Each feed was incubated at 14 g dry matter day−1 with 0, 0.85, 1.7, 3.4, 6.8, 13.6, or 27.2 mg g−1 urea. The amount of urea needed to reach a similar basal concentration of ammonia in the incubation fluid was tested for each feed in advance. Apparent fiber and OM degradability were determined after 48 h of incubation. Data was evaluated by regressions and analysis of variance. The response curve of incubation fluid ammonia to urea supplementation was similar in slope in all feeds. Plateaus in apparent OM degradability in relation to ammonia concentration were determined. The ammonia concentration where apparent OM and fiber degradability reached 95% of maximum was approached in the order of pomace < pulp < hay. With regard to fiber degradability, a plateau was reached at ≥ 80 g kg−1 crude protein only with hay and pomace, whilst a linear relationship existed between RDP and OM degradation for pulp. In hay the ratio RDP: OM degraded was equal to 1.6 but was only 1.0 in the other feeds. There was no obvious lack of branched short-chain fatty acids at low RDP. Thus, the hypothesis was confirmed but the demand for RDP seems even higher in tropical forage compared to food industrial byproducts. The efficiency of urea to promote apparent OM and fiber degradation was also variable. Thus, it seems that minimum thresholds of either RDP or ruminal ammonia concentration may not be reflected appropriately by

  19. Role of live yeasts in rumen ecosystem.

    PubMed

    Oeztuerk, Hakan; Sagmanligil, Vedat

    2009-07-01

    For many years, ruminant nutritionists and microbiologists have been interested in manipulating the microbial ecosystem of the rumen to improve production efficiency by domestic ruminants. Antibiotic ionophores have been used successfully for this purpose. However, the use of antibiotics in animal feeds has been banned in the European Union since January 2006 due to the risk of spreading antibiotic resistance. For this reason, scientists have become interested in evaluating other alternatives to control specific microbial populations to modulate rumen fermentation. Dietary supplements of live yeast preparations, based on Saccharomyces cerevisiae, have been reported to improve health and productivity of ruminants. In contrast to antimicrobial agents, live yeasts offer a natural alternative to manipulate animal performance. This review discusses the modes of action of live yeasts in rumen ecosystem and their subsequent effects on animal performance. PMID:19753793

  20. Development of feeding systems and strategies of supplementation to enhance rumen fermentation and ruminant production in the tropics.

    PubMed

    Wanapat, Metha; Kang, Sungchhang; Polyorach, Sineenart

    2013-01-01

    The availability of local feed resources in various seasons can contribute as essential sources of carbohydrate and protein which significantly impact rumen fermentation and the subsequent productivity of the ruminant. Recent developments, based on enriching protein in cassava chips, have yielded yeast fermented cassava chip protein (YEFECAP) providing up to 47.5% crude protein (CP), which can be used to replace soybean meal. The use of fodder trees has been developed through the process of pelleting; Leucaena leucocephala leaf pellets (LLP), mulberry leaf pellets (MUP) and mangosteen peel and/or garlic pellets, can be used as good sources of protein to supplement ruminant feeding. Apart from producing volatile fatty acids and microbial proteins, greenhouse gases such as methane are also produced in the rumen. Several methods have been used to reduce rumen methane. However, among many approaches, nutritional manipulation using feed formulation and feeding management, especially the use of plant extracts or plants containing secondary compounds (condensed tannins and saponins) and plant oils, has been reported. This approach could help todecrease rumen protozoa and methanogens and thus mitigate the production of methane. At present, more research concerning this burning issue - the role of livestock in global warming - warrants undertaking further research with regard to economic viability and practical feasibility. PMID:23981662

  1. Effect of protein level and urea in concentrate mixture on feed intake and rumen fermentation in swamp buffaloes fed rice straw-based diet.

    PubMed

    Kang, Sungchhang; Wanapat, Metha; Phesatcha, Kampanat; Norrapoke, Thitima

    2015-04-01

    Four rumen-fistulated Thai native swamp buffaloes were randomly assigned according to a 2 × 2 factorial arrangement in a 4 × 4 Latin square design to assess the effect of protein (CP) level and urea (U) source in concentrate diet on feed utilization and rumen ecology. The treatments were as follows: concentrate containing CP at 120 g/kg (soybean meal, SBM) (T1), 160 g/kg (SBM) (T2), 120 g/kg (U) (T3), and 160 g/kg (U) (T4), respectively. All buffaloes were fed concentrate at 10 g/kg of body weight, and rice straw was offered ad libitum. Feed intake and digestibilities of CP, neutral detergent fiber, and acid detergent fiber increased (P < 0.05) in treatments with higher level of CP especially with U source (P < 0.05). In contrast, CP level and source in concentrate did not affect on ruminal pH and temperature (P > 0.05), while concentration of ruminal ammonia (N), blood urea (U), volatile fatty acids profile, microorganism populations, and variable bacterial growth increased in buffaloes consumed concentrate containing CP at 160 g/kg (T2 and T4; P < 0.05). Fecal and urinary N excretions decreased in buffaloes consumed concentrate containing higher CP level especially with U source while purine derivatives increased which resulted in a higher N balance as compared to lower CP level and SBM source treatments (P < 0.05). In summary, higher CP level in concentrate improved feed intake, nutrient digestibility, purine derivatives, and rumen ecology, and U had shown better result than SBM. Concentrate mixtures containing 16 g/kg CP with U 40 g/kg could improved nutrients utilization with no adverse effects for swamp buffaloes fed on rice straw. PMID:25686554

  2. Determining the culturability of the rumen bacterial microbiome.

    PubMed

    Creevey, Christopher J; Kelly, William J; Henderson, Gemma; Leahy, Sinead C

    2014-09-01

    The goal of the Hungate1000 project is to generate a reference set of rumen microbial genome sequences. Toward this goal we have carried out a meta-analysis using information from culture collections, scientific literature, and the NCBI and RDP databases and linked this with a comparative study of several rumen 16S rRNA gene-based surveys. In this way we have attempted to capture a snapshot of rumen bacterial diversity to examine the culturable fraction of the rumen bacterial microbiome. Our analyses have revealed that for cultured rumen bacteria, there are many genera without a reference genome sequence. Our examination of culture-independent studies highlights that there are few novel but many uncultured taxa within the rumen bacterial microbiome. Taken together these results have allowed us to compile a list of cultured rumen isolates that are representative of abundant, novel and core bacterial species in the rumen. In addition, we have identified taxa, particularly within the phylum Bacteroidetes, where further cultivation efforts are clearly required. This information is being used to guide the isolation efforts and selection of bacteria from the rumen microbiota for sequencing through the Hungate1000. PMID:24986151

  3. Determining the culturability of the rumen bacterial microbiome

    PubMed Central

    Creevey, Christopher J; Kelly, William J; Henderson, Gemma; Leahy, Sinead C

    2014-01-01

    The goal of the Hungate1000 project is to generate a reference set of rumen microbial genome sequences. Toward this goal we have carried out a meta-analysis using information from culture collections, scientific literature, and the NCBI and RDP databases and linked this with a comparative study of several rumen 16S rRNA gene-based surveys. In this way we have attempted to capture a snapshot of rumen bacterial diversity to examine the culturable fraction of the rumen bacterial microbiome. Our analyses have revealed that for cultured rumen bacteria, there are many genera without a reference genome sequence. Our examination of culture-independent studies highlights that there are few novel but many uncultured taxa within the rumen bacterial microbiome. Taken together these results have allowed us to compile a list of cultured rumen isolates that are representative of abundant, novel and core bacterial species in the rumen. In addition, we have identified taxa, particularly within the phylum Bacteroidetes, where further cultivation efforts are clearly required. This information is being used to guide the isolation efforts and selection of bacteria from the rumen microbiota for sequencing through the Hungate1000. PMID:24986151

  4. Does Dietary Mitigation of Enteric Methane Production Affect Rumen Function and Animal Productivity in Dairy Cows?

    PubMed Central

    Veneman, Jolien B.; Muetzel, Stefan; Hart, Kenton J.; Faulkner, Catherine L.; Moorby, Jon M.; Perdok, Hink B.; Newbold, Charles J.

    2015-01-01

    It has been suggested that the rumen microbiome and rumen function might be disrupted if methane production in the rumen is decreased. Furthermore concerns have been voiced that geography and management might influence the underlying microbial population and hence the response of the rumen to mitigation strategies. Here we report the effect of the dietary additives: linseed oil and nitrate on methane emissions, rumen fermentation, and the rumen microbiome in two experiments from New Zealand (Dairy 1) and the UK (Dairy 2). Dairy 1 was a randomized block design with 18 multiparous lactating cows. Dairy 2 was a complete replicated 3 x 3 Latin Square using 6 rumen cannulated, lactating dairy cows. Treatments consisted of a control total mixed ration (TMR), supplementation with linseed oil (4% of feed DM) and supplementation with nitrate (2% of feed DM) in both experiments. Methane emissions were measured in open circuit respiration chambers and rumen samples were analyzed for rumen fermentation parameters and microbial population structure using qPCR and next generation sequencing (NGS). Supplementation with nitrate, but not linseed oil, decreased methane yield (g/kg DMI; P<0.02) and increased hydrogen (P<0.03) emissions in both experiments. Furthermore, the effect of nitrate on gaseous emissions was accompanied by an increased rumen acetate to propionate ratio and consistent changes in the rumen microbial populations including a decreased abundance of the main genus Prevotella and a decrease in archaeal mcrA (log10 copies/ g rumen DM content). These results demonstrate that methane emissions can be significantly decreased with nitrate supplementation with only minor, but consistent, effects on the rumen microbial population and its function, with no evidence that the response to dietary additives differed due to geography and different underlying microbial populations. PMID:26509835

  5. Does Dietary Mitigation of Enteric Methane Production Affect Rumen Function and Animal Productivity in Dairy Cows?

    PubMed

    Veneman, Jolien B; Muetzel, Stefan; Hart, Kenton J; Faulkner, Catherine L; Moorby, Jon M; Perdok, Hink B; Newbold, Charles J

    2015-01-01

    It has been suggested that the rumen microbiome and rumen function might be disrupted if methane production in the rumen is decreased. Furthermore concerns have been voiced that geography and management might influence the underlying microbial population and hence the response of the rumen to mitigation strategies. Here we report the effect of the dietary additives: linseed oil and nitrate on methane emissions, rumen fermentation, and the rumen microbiome in two experiments from New Zealand (Dairy 1) and the UK (Dairy 2). Dairy 1 was a randomized block design with 18 multiparous lactating cows. Dairy 2 was a complete replicated 3 x 3 Latin Square using 6 rumen cannulated, lactating dairy cows. Treatments consisted of a control total mixed ration (TMR), supplementation with linseed oil (4% of feed DM) and supplementation with nitrate (2% of feed DM) in both experiments. Methane emissions were measured in open circuit respiration chambers and rumen samples were analyzed for rumen fermentation parameters and microbial population structure using qPCR and next generation sequencing (NGS). Supplementation with nitrate, but not linseed oil, decreased methane yield (g/kg DMI; P<0.02) and increased hydrogen (P<0.03) emissions in both experiments. Furthermore, the effect of nitrate on gaseous emissions was accompanied by an increased rumen acetate to propionate ratio and consistent changes in the rumen microbial populations including a decreased abundance of the main genus Prevotella and a decrease in archaeal mcrA (log10 copies/g rumen DM content). These results demonstrate that methane emissions can be significantly decreased with nitrate supplementation with only minor, but consistent, effects on the rumen microbial population and its function, with no evidence that the response to dietary additives differed due to geography and different underlying microbial populations. PMID:26509835

  6. Does Dietary Mitigation of Enteric Methane Production Affect Rumen Function and Animal Productivity in Dairy Cows?

    PubMed

    Veneman, Jolien B; Muetzel, Stefan; Hart, Kenton J; Faulkner, Catherine L; Moorby, Jon M; Perdok, Hink B; Newbold, Charles J

    2015-01-01

    It has been suggested that the rumen microbiome and rumen function might be disrupted if methane production in the rumen is decreased. Furthermore concerns have been voiced that geography and management might influence the underlying microbial population and hence the response of the rumen to mitigation strategies. Here we report the effect of the dietary additives: linseed oil and nitrate on methane emissions, rumen fermentation, and the rumen microbiome in two experiments from New Zealand (Dairy 1) and the UK (Dairy 2). Dairy 1 was a randomized block design with 18 multiparous lactating cows. Dairy 2 was a complete replicated 3 x 3 Latin Square using 6 rumen cannulated, lactating dairy cows. Treatments consisted of a control total mixed ration (TMR), supplementation with linseed oil (4% of feed DM) and supplementation with nitrate (2% of feed DM) in both experiments. Methane emissions were measured in open circuit respiration chambers and rumen samples were analyzed for rumen fermentation parameters and microbial population structure using qPCR and next generation sequencing (NGS). Supplementation with nitrate, but not linseed oil, decreased methane yield (g/kg DMI; P<0.02) and increased hydrogen (P<0.03) emissions in both experiments. Furthermore, the effect of nitrate on gaseous emissions was accompanied by an increased rumen acetate to propionate ratio and consistent changes in the rumen microbial populations including a decreased abundance of the main genus Prevotella and a decrease in archaeal mcrA (log10 copies/g rumen DM content). These results demonstrate that methane emissions can be significantly decreased with nitrate supplementation with only minor, but consistent, effects on the rumen microbial population and its function, with no evidence that the response to dietary additives differed due to geography and different underlying microbial populations.

  7. Detect changes in protein structure of carinata meal during rumen fermentation in relation to basic chemical profile and comparison with canola meal using ATR-FT/IR molecular spectroscopy with chemometrics

    NASA Astrophysics Data System (ADS)

    Xin, Hangshu; Yu, Peiqiang

    2013-08-01

    As far as we know, no study has been carried out on whether protein structure changes in the feed during rumen fermentation from other research team. This study was conducted to characterize protein structure spectral changes in carinata meal during ruminal fermentation using Fourier transform infrared spectroscopy (FT/IR) technique with ATR. The objectives were to find out whether (1) protein internal structure (in terms of protein amide profile and protein secondary structure profile) changed after in situ ruminal fermentation at 0, 12, 24 and 48 h in carinata meal and conventional canola meal was used as a reference; (2) there was any correlation between protein spectral parameters and basic chemical profile in in situ rumen residue samples; and (3) the protein structural chemical make-up of carinata meal differed from canola meal during 48 h rumen incubation. The results showed that protein structure features in both carinata meal and canola meal were altered as incubation time increased (P < 0.0001) and linear and curvilinear relationships (P < 0.05) on amide II height and area, height and area ratio of amide I and II as well as height ratio of α-helix and β-sheet were observed within 48 h ruminal fermentation. And the amide I height and area as well as α-helix height and β-sheet height were in the highest level of IR absorbance at 0 h and then gradually declined linearly (P < 0.0001) by 30-38% after 48 h incubation. These results indicated that not only quantities decreased but also inherent structure changed in protein chemical make-up during ruminal fermentation. Meanwhile, strong correlations were found between protein spectral parameters and some basic nutrients profile such as CP (positively) and NDF (negatively). And both AHCA and PCA results showed that in situ rumen residues from carinata meal was not distinguished from those from canola meal, suggesting some relationship in structural make-up exhibited between them within protein region during 48

  8. An Integrated Multi-Omics Approach Reveals the Effects of Supplementing Grass or Grass Hay with Vitamin E on the Rumen Microbiome and Its Function

    PubMed Central

    Belanche, Alejandro; Kingston-Smith, Alison H.; Newbold, Charles J.

    2016-01-01

    Rumen function is generally suboptimal leading to losses in methane and nitrogen. Analysis of the rumen microbiome is thus important to understanding the underlying microbial activity under different feeding strategies. This study investigated the effect of forage conservation method and vitamin E supplementation on rumen function using a rumen simulation technique. Ryegrass (GRA) or ryegrass hay (HAY) was supplemented with 20% concentrate containing zero or 50 IU/d vitamin E, as α-tocopheryl acetate, according to a 2 × 2 factorial design. The forage conservation method did not substantially change the nutrient composition but had a profound impact on the structure and diversity of the rumen microbiome. HAY diets promoted a more complex bacterial community (+38 OTUs) dominated by Firmicutes. This bacterial adaptation, together with increased rumen protozoa levels and methanogen diversity, was associated with greater fiber disappearance (+12%) in HAY diets, but also with greater rumen true N degradability (+7%) than GRA diets. HAY diets also had a higher metabolic H recovery and methane production (+35%) suggesting more efficient inter-species H transfer between bacteria, protozoa and methanogens. Contrarily, GRA diets promoted more simplified methanogen and bacterial communities, which were dominated by Bacteroidetes and Lactobacillus, thus lactate formation may have acted as an alternative H sink in GRA diets. Moreover the structure of the bacterial community with GRA diets was highly correlated with N utilization, and GRA diets promoted greater bacterial growth and microbial protein synthesis (+16%), as well as a more efficient microbial protein synthesis (+22%). A dose-response experiment using batch cultures revealed that vitamin E supplementation increased rumen fermentation in terms of total VFA and gas production, with protozoal activity higher when supplying α-tocopheryl acetate vs. α-tocopherol. Moreover, α-tocopheryl acetate promoted a small increase

  9. An Integrated Multi-Omics Approach Reveals the Effects of Supplementing Grass or Grass Hay with Vitamin E on the Rumen Microbiome and Its Function.

    PubMed

    Belanche, Alejandro; Kingston-Smith, Alison H; Newbold, Charles J

    2016-01-01

    Rumen function is generally suboptimal leading to losses in methane and nitrogen. Analysis of the rumen microbiome is thus important to understanding the underlying microbial activity under different feeding strategies. This study investigated the effect of forage conservation method and vitamin E supplementation on rumen function using a rumen simulation technique. Ryegrass (GRA) or ryegrass hay (HAY) was supplemented with 20% concentrate containing zero or 50 IU/d vitamin E, as α-tocopheryl acetate, according to a 2 × 2 factorial design. The forage conservation method did not substantially change the nutrient composition but had a profound impact on the structure and diversity of the rumen microbiome. HAY diets promoted a more complex bacterial community (+38 OTUs) dominated by Firmicutes. This bacterial adaptation, together with increased rumen protozoa levels and methanogen diversity, was associated with greater fiber disappearance (+12%) in HAY diets, but also with greater rumen true N degradability (+7%) than GRA diets. HAY diets also had a higher metabolic H recovery and methane production (+35%) suggesting more efficient inter-species H transfer between bacteria, protozoa and methanogens. Contrarily, GRA diets promoted more simplified methanogen and bacterial communities, which were dominated by Bacteroidetes and Lactobacillus, thus lactate formation may have acted as an alternative H sink in GRA diets. Moreover the structure of the bacterial community with GRA diets was highly correlated with N utilization, and GRA diets promoted greater bacterial growth and microbial protein synthesis (+16%), as well as a more efficient microbial protein synthesis (+22%). A dose-response experiment using batch cultures revealed that vitamin E supplementation increased rumen fermentation in terms of total VFA and gas production, with protozoal activity higher when supplying α-tocopheryl acetate vs. α-tocopherol. Moreover, α-tocopheryl acetate promoted a small increase

  10. The giraffe (Giraffa camelopardalis) rumen microbiome.

    PubMed

    Roggenbuck, Michael; Sauer, Cathrine; Poulsen, Morten; Bertelsen, Mads F; Sørensen, Søren J

    2014-10-01

    Recent studies have shown that wild ruminants are sources of previously undescribed microorganisms, knowledge of which can improve our understanding of the complex microbial interactions in the foregut. Here, we investigated the microbial community of seven wild-caught giraffes (Giraffa camelopardalis), three of which were fed natural browse and four were fed Boskos pellets, leafy alfalfa hay, and cut savanna browse, by characterizing the 16S rRNA gene diversity using 454 FLX high-throughput sequencing. The microbial community composition varied according to diet, but differed little between the ruminal fluid and solid fraction. The giraffe rumen contained large levels of the phyla of Firmicutes and Bacteroidetes independent of diet, while Prevotella, Succinclasticium, and Methanobrevibacter accounted for the largest abundant taxonomic assigned genera. However, up to 21% of the generated sequences could not been assigned to any known bacterial phyla, and c. 70% not to genus, revealing that the giraffe rumen hosts a variety of previously undescribed bacteria.

  11. The giraffe (Giraffa camelopardalis) rumen microbiome.

    PubMed

    Roggenbuck, Michael; Sauer, Cathrine; Poulsen, Morten; Bertelsen, Mads F; Sørensen, Søren J

    2014-10-01

    Recent studies have shown that wild ruminants are sources of previously undescribed microorganisms, knowledge of which can improve our understanding of the complex microbial interactions in the foregut. Here, we investigated the microbial community of seven wild-caught giraffes (Giraffa camelopardalis), three of which were fed natural browse and four were fed Boskos pellets, leafy alfalfa hay, and cut savanna browse, by characterizing the 16S rRNA gene diversity using 454 FLX high-throughput sequencing. The microbial community composition varied according to diet, but differed little between the ruminal fluid and solid fraction. The giraffe rumen contained large levels of the phyla of Firmicutes and Bacteroidetes independent of diet, while Prevotella, Succinclasticium, and Methanobrevibacter accounted for the largest abundant taxonomic assigned genera. However, up to 21% of the generated sequences could not been assigned to any known bacterial phyla, and c. 70% not to genus, revealing that the giraffe rumen hosts a variety of previously undescribed bacteria. PMID:25087453

  12. [Effect of physical treatment of rapeseed expeller, wheat, corn and corn gluten feed on the degradability in the rumen and the enzymatic in vitro digestibility of nondegraded crude proteins].

    PubMed

    Sommer, A; Chrenková, M; Ceresnaková, Z; Peisker, M

    1994-01-01

    The influence of physical treatment-expansion and flaking-on crude proteins degradability in the rumen was studied in maize, maize-gluten feed, rape extracted meal and in the expanded one at 120 degrees C and 150 degrees C, rape cake, wheat and flaked wheat by in sacco method. The enzymatic digestibility of crude protein in the rumen undegraded residues of the above mentioned feeds was determined by an enzymatically in vitro method. The treatment of feed decreased significantly the original solubility and theoretical degradability of crude proteins, and the amount of undegraded crude proteins was increased. Positive influence on the amount of enzymatically digested crude protein was determined in rape expanded at 120 degrees C and 150 degrees C (60, 61 and/or 68%). Flaking of wheat had a similar effect. Enzymatic digestibility at undegraded rests where increased by 8-10% after the heat treatment and it remained almost unchanged in expanded maize-gluten feed. PMID:7717847

  13. Potential opportunities and problems for genetically altered rumen microorganisms.

    PubMed

    Russell, J B; Wilson, D B

    1988-02-01

    Rumen microbiologists are beginning to use genetic engineering techniques, and researchers should carefully consider both the potentials and limitations of using this technology to manipulate the rumen microbial ecosystem. Despite encouraging rhetoric, it is difficult to identify specific examples where genetic engineering would enhance ruminal performance. Many practical problems (lactic acidosis, deamination, etc.) might be better served by genetic engineering approaches that delete rather than add genes. The difficulty with this approach is that a highly selective means of preventing wild types from recolonizing the rumen would be needed. The addition of specific genes is confounded by 1) the fact that the rumen microorganisms are already adapted to the rumen, 2) the diversity of species inhabiting the rumen and 3) the complexity of interactions among these species. Aspects such as increased rates of cellulose digestion and changes in amino acid composition of the microflora are particularly sensitive to these biological constraints. Genetic engineering has, however, the potential to alleviate new limitations that humans have imposed on the rumen (detoxification, resistance to low pH, the digestion of novel feed materials, etc). A particular strategy of moving acid-resistant cellulose genes into noncellulytic, but acid-resistant, rumen bacteria is described.

  14. Rumen-protected methionine and lysine: effects on milk production and plasma amino acids of dairy cows with reference to metabolisable protein status.

    PubMed

    Awawdeh, Mofleh S

    2016-05-01

    Two experiments were conducted to study the effects of rumen-protected Met (RPM) alone or with rumen-protected Lys (RPL) on milk yield and plasma amino acids of dairy cows. In experiment 1, 24 multiparous Holstein cows (154 DIM) were assigned to one of 3 groups where each cow received 0 g/d of RPM and RPL (C), 30 g/d of RPM (M), or 30 g/d of RPM plus 25 g of RPL (ML). The study lasted for 8 weeks where milk yield and composition were determined weekly. Daily milk yield averaged 28·0, 27·8, and 29·7 kg/cow for the C, M, and ML groups, respectively. Dietary treatments had no effects (P ≥ 0·54) on milk contents of fat, lactose, solid non-fat or total solids. Milk protein content in the ML group was greater (P < 0·05) than the C and M groups. Plasma levels of all AA were not significantly (P ≥ 0·09) affected by supplemental RPL and/or RPM. In experiment 2, 30 multiparous Holstein cows (100 DIM) were assigned to one of 3 groups where each cow received 0 g/d of RPM and RPL (C), 50 g/d of RPM (M), or 50 g/d of RPM plus 25 g/d of RPL (ML). The study lasted for 5 weeks. Cows in the M (30·5 kg) and ML (31·4 kg) groups produced (P < 0·05) more milk than those of the C group (29·1 kg). Under conditions of this study, RPM plus RPL improved milk yield and protein contents of dairy cows and was better than supplying RPM alone. Response in milk yield to RPM and RPL was affected by the MP status of cows which deserves further investigation.

  15. Role and function of short chain fatty acids in rumen epithelial metabolism, development and importance of the rumen epithelium in understanding control of transcriptome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The epithelial lining of the rumen is uniquely placed to have impact on the nutrient metabolism of the animal. The symbiotic relationship with the microbial populations that inhabit the rumen, serves to provide a constant supply of nutrients from roughage that would otherwise be unusable. Metaboli...

  16. Rumen microbiome from steers differing in feed efficiency

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle rumen has a diverse microbial ecosystem that is essential for the host to digest plant material. Extremes in body weight (BW) gain in mice and humans have been associated with different intestinal microbial populations. The objective of this study was to characterize the microbiome of th...

  17. Influence of temperature and humidity on rumen pH and fatty acids in dairy cows.

    PubMed

    Gianesella, M; Piccione, G; Cannizzo, C; Casella, S; Morgante, M

    2012-11-01

    The aim of this study was to investigate the variations of rumen pH and fatty acids (acetic acid, propionic acid, iso-butyric acid, n-butyric acid, iso-valerianic acid, n-valerianic, caproic acid and total fatty acids) in 245 early lactating dairy cows under different temperature and humidity conditions. The animals were divided into six groups and rumen fluid was collected by rumenocentesis on 22 dairy cows in April (Group A), 33 in May (Group B), 43 in June (Group C), 48 in July (Group D), 36 in September (Group E) and 60 in October (Group F). One-way analysis of variance (ANOVA), followed by the Bonferroni's test, showed a significant effect of environmental variations on all studied parameters (P < 0.0001). Changes in studied parameters can be explained in relation to the microbial population and shift in the optima for rumen conditions associated with variations of environmental conditions. We can affirm that the microbial assemblages that underlie energy and protein supply to wild ruminant are evident especially in relation to temperature and humidity conditions.

  18. Symbiosis and Rumen Protozoa

    ERIC Educational Resources Information Center

    Dillon, Raymond D.

    1970-01-01

    Protozoa inhabiting the rumen of large grazing animals can be used to illustrate symbiotic animal associations. Gives a key to the ciliates most commonly found, several drawings, and a chart relating rumen fauna to the phylogenetic tree of the hosts. (EB)

  19. Effects of Supplementation of Eucalyptus (E. Camaldulensis) Leaf Meal on Feed Intake and Rumen Fermentation Efficiency in Swamp Buffaloes

    PubMed Central

    Thao, N. T.; Wanapat, M.; Kang, S.; Cherdthong, A.

    2015-01-01

    Four rumen fistulated swamp buffaloes were randomly assigned according to a 4×4 Latin square design to investigate the effects of Eucalyptus (E. Camaldulensis) leaf meal (ELM) supplementation as a rumen enhancer on feed intake and rumen fermentation characteristics. The dietary treatments were as follows: T1 = 0 g ELM/hd/d; T2 = 40 g ELM/hd/d; T3 = 80 g ELM/hd/d; T4 = 120 g ELM/hd/d, respectively. Experimental animals were kept in individual pens and concentrate was offered at 0.3% BW while rice straw was fed ad libitum. The results revealed that voluntary feed intake and digestion coefficients of nutrients were similar among treatments. Ruminal pH, temperature and blood urea nitrogen concentrations were not affected by ELM supplementation; however, ELM supplementation resulted in lower concentration of ruminal ammonia nitrogen. Total volatile fatty acids, propionate concentration increased with the increasing level of EML (p<0.05) while the proportion of acetate was decreased (p<0.05). Methane production was linearly decreased (p<0.05) with the increasing level of ELM supplementation. Protozoa count and proteolytic bacteria population were reduced (p<0.05) while fungal zoospores and total viable bacteria, amylolytic, cellulolytic bacteria were unchanged. In addition, nitrogen utilization and microbial protein synthesis tended to increase by the dietary treatments. Based on the present findings, it is suggested that ELM could modify the rumen fermentation and is potentially used as a rumen enhancer in methane mitigation and rumen fermentation efficiency. PMID:26104399

  20. Insights into Abundant Rumen Ureolytic Bacterial Community Using Rumen Simulation System.

    PubMed

    Jin, Di; Zhao, Shengguo; Wang, Pengpeng; Zheng, Nan; Bu, Dengpan; Beckers, Yves; Wang, Jiaqi

    2016-01-01

    Urea, a non-protein nitrogen for dairy cows, is rapidly hydrolyzed to ammonia by urease produced by ureolytic bacteria in the rumen, and the ammonia is used as nitrogen for rumen bacterial growth. However, there is limited knowledge with regard to the ureolytic bacteria community in the rumen. To explore the ruminal ureolytic bacterial community, urea, or acetohydroxamic acid (AHA, an inhibitor of urea hydrolysis) were supplemented into the rumen simulation systems. The bacterial 16S rRNA genes were sequenced by Miseq high-throughput sequencing and used to reveal the ureoltyic bacteria by comparing different treatments. The results revealed that urea supplementation significantly increased the ammonia concentration, and AHA addition inhibited urea hydrolysis. Urea supplementation significantly increased the richness of bacterial community and the proportion of ureC genes. The composition of bacterial community following urea or AHA supplementation showed no significant difference compared to the groups without supplementation. The abundance of Bacillus and unclassified Succinivibrionaceae increased significantly following urea supplementation. Pseudomonas, Haemophilus, Neisseria, Streptococcus, and Actinomyces exhibited a positive response to urea supplementation and a negative response to AHA addition. Results retrieved from the NCBI protein database and publications confirmed that the representative bacteria in these genera mentioned above had urease genes or urease activities. Therefore, the rumen ureolytic bacteria were abundant in the genera of Pseudomonas, Haemophilus, Neisseria, Streptococcus, Actinomyces, Bacillus, and unclassified Succinivibrionaceae. Insights into abundant rumen ureolytic bacteria provide the regulation targets to mitigate urea hydrolysis and increase efficiency of urea nitrogen utilization in ruminants. PMID:27446045

  1. Insights into Abundant Rumen Ureolytic Bacterial Community Using Rumen Simulation System

    PubMed Central

    Jin, Di; Zhao, Shengguo; Wang, Pengpeng; Zheng, Nan; Bu, Dengpan; Beckers, Yves; Wang, Jiaqi

    2016-01-01

    Urea, a non-protein nitrogen for dairy cows, is rapidly hydrolyzed to ammonia by urease produced by ureolytic bacteria in the rumen, and the ammonia is used as nitrogen for rumen bacterial growth. However, there is limited knowledge with regard to the ureolytic bacteria community in the rumen. To explore the ruminal ureolytic bacterial community, urea, or acetohydroxamic acid (AHA, an inhibitor of urea hydrolysis) were supplemented into the rumen simulation systems. The bacterial 16S rRNA genes were sequenced by Miseq high-throughput sequencing and used to reveal the ureoltyic bacteria by comparing different treatments. The results revealed that urea supplementation significantly increased the ammonia concentration, and AHA addition inhibited urea hydrolysis. Urea supplementation significantly increased the richness of bacterial community and the proportion of ureC genes. The composition of bacterial community following urea or AHA supplementation showed no significant difference compared to the groups without supplementation. The abundance of Bacillus and unclassified Succinivibrionaceae increased significantly following urea supplementation. Pseudomonas, Haemophilus, Neisseria, Streptococcus, and Actinomyces exhibited a positive response to urea supplementation and a negative response to AHA addition. Results retrieved from the NCBI protein database and publications confirmed that the representative bacteria in these genera mentioned above had urease genes or urease activities. Therefore, the rumen ureolytic bacteria were abundant in the genera of Pseudomonas, Haemophilus, Neisseria, Streptococcus, Actinomyces, Bacillus, and unclassified Succinivibrionaceae. Insights into abundant rumen ureolytic bacteria provide the regulation targets to mitigate urea hydrolysis and increase efficiency of urea nitrogen utilization in ruminants. PMID:27446045

  2. Parameters of fermentation and rumen microbiota of Nellore steers fed with different proportions of concentrate in fresh sugarcane containing diets.

    PubMed

    Ribeiro Júnior, Carlos S; Messana, Juliana D; Granja-Salcedo, Yury T; Canesin, Roberta C; Fiorentini, Giovani; San Vito, Elias; Furlan, Luiz R; Reis, Ricardo A; Berchielli, Telma T

    2016-10-01

    The objective of this study was to evaluate the effect of a fresh sugarcane-based diet and different roughage-to-concentrate ratios (70:30, 60:40, 40:60 and 20:80) on the rumen microbiota associated with rumen fermentation parameters and the intake and apparent digestibility of nutrients in Nellore steers. Eight rumen-cannulated Nellore steers (331 ± 8 kg BW) were distributed in a double 4 × 4 Latin square design balanced for the control of the residual effect. The ruminal pH decreased (p < 0.01) and the concentrations of N-NH3, isovaleric and valeric acids increased linearly (p < 0.05) with an increase dietary concentrate level. Furthermore, an increased concentrate proportion reduced the population of Fibrobacter succinogenes and Ruminococus flavefaciens (p < 0.01) and increased the population of Selenomonas ruminantium and Megasphaera elsdenii (p < 0.01). The protozoa count revealed a predominance of the genus Entodinium. The synthesis of microbial N [g/d] and the efficiency of microbial synthesis [g of microbial N/kg of organic matter apparently digested in the rumen] increased as the proportion of concentrate was increased (p < 0.05). Therefore, it can be concluded that an increasing proportion of concentrate in sugarcane-containing diets enhances the synthesis of microbial protein and does not alter the fibre digestibility, although the population of fibre fermenting bacteria was reduced. PMID:27415825

  3. Variation of in situ rumen degradation of crude protein and amino acids and in vitro digestibility of undegraded feed protein in rapeseed meals.

    PubMed

    Steingass, H; Kneer, G; Wischer, G; Rodehutscord, M

    2013-07-01

    In this study, 10 samples of rapeseed meal (RSM) from 10 different oil plants in Germany were examined. In situ rumen degradation of CP was determined by incubation over 1, 2, 4, 8, 16, 32 and 72 h in duplicate per time point using three rumen fistulated dry cows. Degradation kinetics were estimated by an exponential model and effective CP degradation was calculated. Degradation was corrected for small particle loss as the difference between washing loss and water-soluble fraction. Amino acid analysis was carried out in the samples and in the residues after 8 and 16 h of incubation in situ and degradation of individual amino acids was calculated for these incubation times. In vitro pepsin-pancreatin digestibility of CP (IPD) was determined in the samples as well as in the 8 and 16 h residues. Effective CP degradation for a rumen outflow rate of 8%/h (ED8) averaged 54.3% with a considerable variation among samples ranging from 44.3% to 62.7%. A multiple regression equation containing acid detergent insoluble N, total glucosinolates and petroleum ether extract as independent variables predicted ED8 with satisfying accuracy (R 2 = 0.74; RSD = 6.4%). Degradation of amino acids was different from that of CP for most amino acids studied, especially after 8 h of incubation. Compared with CP, degradation of essential amino acids was predominantly lower while degradation of non-essential amino acids was higher in most cases. However, for lysine and methionine no distinct difference with CP degradation was found. Degradation of individual amino acids was predicted from CP degradation with high accuracy using linear regression equations. Average IPD of RSM was 79.8 ± 2.6%. IPD was lower in the incubation residues and decreased with longer incubation time and increasing rumen degradation, respectively.

  4. Variation of in situ rumen degradation of crude protein and amino acids and in vitro digestibility of undegraded feed protein in rapeseed meals.

    PubMed

    Steingass, H; Kneer, G; Wischer, G; Rodehutscord, M

    2013-07-01

    In this study, 10 samples of rapeseed meal (RSM) from 10 different oil plants in Germany were examined. In situ rumen degradation of CP was determined by incubation over 1, 2, 4, 8, 16, 32 and 72 h in duplicate per time point using three rumen fistulated dry cows. Degradation kinetics were estimated by an exponential model and effective CP degradation was calculated. Degradation was corrected for small particle loss as the difference between washing loss and water-soluble fraction. Amino acid analysis was carried out in the samples and in the residues after 8 and 16 h of incubation in situ and degradation of individual amino acids was calculated for these incubation times. In vitro pepsin-pancreatin digestibility of CP (IPD) was determined in the samples as well as in the 8 and 16 h residues. Effective CP degradation for a rumen outflow rate of 8%/h (ED8) averaged 54.3% with a considerable variation among samples ranging from 44.3% to 62.7%. A multiple regression equation containing acid detergent insoluble N, total glucosinolates and petroleum ether extract as independent variables predicted ED8 with satisfying accuracy (R 2 = 0.74; RSD = 6.4%). Degradation of amino acids was different from that of CP for most amino acids studied, especially after 8 h of incubation. Compared with CP, degradation of essential amino acids was predominantly lower while degradation of non-essential amino acids was higher in most cases. However, for lysine and methionine no distinct difference with CP degradation was found. Degradation of individual amino acids was predicted from CP degradation with high accuracy using linear regression equations. Average IPD of RSM was 79.8 ± 2.6%. IPD was lower in the incubation residues and decreased with longer incubation time and increasing rumen degradation, respectively. PMID:23472609

  5. The rumen plasmidome: A genetic communication hub for the rumen microbiome.

    PubMed

    Mizrahi, Itzhak

    2012-05-01

    Plasmids are episomally replicating genetic elements which carry backbone genes that are important for their replication and maintenance within their host, and accessory genes that might confer an advantage to their host in its ecological niche. As such, they are often perceived as a powerful evolutionary force, which horizontally introduces new traits into bacterial cells and genomes. In our recent publication "Insight into the rumen plasmidome" we characterized the metagenomic plasmid population of the bovine rumen microbial ecological niche. The rumen is the first compartment of the digestive tract of ruminants; it functions as a pre-gastric anaerobic fermentation chamber, where plant fibers are degraded and converted into chemical compounds which are subsequently absorbed and digested by the animal. PMID:23061023

  6. Effects of different tannin-rich extracts and rapeseed tannin monomers on methane formation and microbial protein synthesis in vitro.

    PubMed

    Wischer, G; Boguhn, J; Steingaß, H; Schollenberger, M; Rodehutscord, M

    2013-11-01

    Tannins, polyphenolic compounds found in plants, are known to complex with proteins of feed and rumen bacteria. This group of substances has the potential to reduce methane production either with or without negative effects on digestibility and microbial yield. In the first step of this study, 10 tannin-rich extracts from chestnut, mimosa, myrabolan, quebracho, sumach, tara, valonea, oak, cocoa and grape seed, and four rapeseed tannin monomers (pelargonidin, catechin, cyanidin and sinapinic acid) were used in a series of in vitro trials using the Hohenheim gas test, with grass silage as substrate. The objective was to screen the potential of various tannin-rich extracts to reduce methane production without a significant effect on total gas production (GP). Supplementation with pelargonidin and cyanidin did not reduce methane production; however, catechin and sinapinic acid reduced methane production without altering GP. All tannin-rich extracts, except for tara extract, significantly reduced methane production by 8% to 28% without altering GP. On the basis of these results, five tannin-rich extracts were selected and further investigated in a second step using a Rusitec system. Each tannin-rich extract (1.5 g) was supplemented to grass silage (15 g). In this experiment, nutrient degradation, microbial protein synthesis and volatile fatty acid production were used as additional response criteria. Chestnut extract caused the greatest reduction in methane production followed by valonea, grape seed and sumach, whereas myrabolan extract did not reduce methane production. Whereas chestnut extract reduced acetate production by 19%, supplementation with grape seed or myrabolan extract increased acetate production. However, degradation of fibre fractions was reduced in all tannin treatments. Degradation of dry matter and organic matter was also reduced by tannin supplementation, and no differences were found between the tannin-rich extracts. CP degradation and ammonia

  7. An adhesin from hydrogen-utilizing rumen methanogen Methanobrevibacter ruminantium M1 binds a broad range of hydrogen-producing microorganisms.

    PubMed

    Ng, Filomena; Kittelmann, Sandra; Patchett, Mark L; Attwood, Graeme T; Janssen, Peter H; Rakonjac, Jasna; Gagic, Dragana

    2016-09-01

    Symbiotic associations are ubiquitous in the microbial world and have a major role in shaping the evolution of both partners. One of the most interesting mutualistic relationships exists between protozoa and methanogenic archaea in the fermentative forestomach (rumen) of ruminant animals. Methanogens reside within and on the surface of protozoa as symbionts, and interspecies hydrogen transfer is speculated to be the main driver for physical associations observed between the two groups. In silico analyses of several rumen methanogen genomes have previously shown that up to 5% of genes encode adhesin-like proteins, which may be central to rumen interspecies attachment. We hypothesized that adhesin-like proteins on methanogen cell surfaces facilitate attachment to protozoal hosts. Using phage display technology, we have identified a protein (Mru_1499) from Methanobrevibacter ruminantium M1 as an adhesin that binds to a broad range of rumen protozoa (including the genera Epidinium and Entodinium). This unique adhesin also binds the cell surface of the bacterium Butyrivibrio proteoclasticus, suggesting a broad adhesion spectrum for this protein.

  8. An adhesin from hydrogen-utilizing rumen methanogen Methanobrevibacter ruminantium M1 binds a broad range of hydrogen-producing microorganisms.

    PubMed

    Ng, Filomena; Kittelmann, Sandra; Patchett, Mark L; Attwood, Graeme T; Janssen, Peter H; Rakonjac, Jasna; Gagic, Dragana

    2016-09-01

    Symbiotic associations are ubiquitous in the microbial world and have a major role in shaping the evolution of both partners. One of the most interesting mutualistic relationships exists between protozoa and methanogenic archaea in the fermentative forestomach (rumen) of ruminant animals. Methanogens reside within and on the surface of protozoa as symbionts, and interspecies hydrogen transfer is speculated to be the main driver for physical associations observed between the two groups. In silico analyses of several rumen methanogen genomes have previously shown that up to 5% of genes encode adhesin-like proteins, which may be central to rumen interspecies attachment. We hypothesized that adhesin-like proteins on methanogen cell surfaces facilitate attachment to protozoal hosts. Using phage display technology, we have identified a protein (Mru_1499) from Methanobrevibacter ruminantium M1 as an adhesin that binds to a broad range of rumen protozoa (including the genera Epidinium and Entodinium). This unique adhesin also binds the cell surface of the bacterium Butyrivibrio proteoclasticus, suggesting a broad adhesion spectrum for this protein. PMID:26643468

  9. Effects of dietary crude protein and rumen-degradable protein concentrations on urea recycling, nitrogen balance, omasal nutrient flow, and milk production in dairy cows.

    PubMed

    Mutsvangwa, T; Davies, K L; McKinnon, J J; Christensen, D A

    2016-08-01

    The objective of this study was to determine how interactions between dietary crude protein (CP) and rumen-degradable protein (RDP) concentrations alter urea-nitrogen recycling, nitrogen (N) balance, omasal nutrient flow, and milk production in lactating Holstein cows. Eight multiparous Holstein cows (711±21kg of body weight; 91±17d in milk at the start of the experiment) were used in a replicated 4×4 Latin square design with a 2×2 factorial arrangement of dietary treatments and 29-d experimental periods. Four cows in one Latin square were fitted with ruminal cannulas to allow ruminal and omasal sampling. The dietary treatment factors were CP (14.9 vs. 17.5%; dry matter basis) and RDP (63 vs. 69% of CP) contents. Dietary RDP concentration was manipulated by including unprocessed or micronized canola meal. Diet adaptation (d 1-20) was followed by 8d (d 21-29) of sample and data collection. Continuous intrajugular infusions of [(15)N(15)N]-urea (220mg/d) were conducted for 4d (d 25-29) with concurrent total collections of urine and feces to estimate N balance and whole-body urea kinetics. Proportions of [(15)N(15)N]- and [(14)N(15)N]-urea in urinary urea, and (15)N enrichment in feces were used to calculate urea kinetics. For the low-CP diets, cows fed the high-RDP diet had a greater DM intake compared with those fed the low-RDP diet, but the opposite trend was observed for cows fed the high-CP diets. Dietary treatment had no effect on milk yield. Milk composition and milk component yields were largely unaffected by dietary treatment; however, on the low-CP diets, milk fat yield was greater for cows fed the low-RDP diet compared with those fed the high-RDP diet, but it was unaffected by RDP concentration on the high-CP diets. On the high-CP diets, milk urea nitrogen concentration was greater in cows fed the high-RDP diet compared with those fed the low-RDP diet, but it was unaffected by RDP concentration on the low-CP diets. Ruminal NH3-N concentration tended to

  10. Effects of dietary crude protein and rumen-degradable protein concentrations on urea recycling, nitrogen balance, omasal nutrient flow, and milk production in dairy cows.

    PubMed

    Mutsvangwa, T; Davies, K L; McKinnon, J J; Christensen, D A

    2016-08-01

    The objective of this study was to determine how interactions between dietary crude protein (CP) and rumen-degradable protein (RDP) concentrations alter urea-nitrogen recycling, nitrogen (N) balance, omasal nutrient flow, and milk production in lactating Holstein cows. Eight multiparous Holstein cows (711±21kg of body weight; 91±17d in milk at the start of the experiment) were used in a replicated 4×4 Latin square design with a 2×2 factorial arrangement of dietary treatments and 29-d experimental periods. Four cows in one Latin square were fitted with ruminal cannulas to allow ruminal and omasal sampling. The dietary treatment factors were CP (14.9 vs. 17.5%; dry matter basis) and RDP (63 vs. 69% of CP) contents. Dietary RDP concentration was manipulated by including unprocessed or micronized canola meal. Diet adaptation (d 1-20) was followed by 8d (d 21-29) of sample and data collection. Continuous intrajugular infusions of [(15)N(15)N]-urea (220mg/d) were conducted for 4d (d 25-29) with concurrent total collections of urine and feces to estimate N balance and whole-body urea kinetics. Proportions of [(15)N(15)N]- and [(14)N(15)N]-urea in urinary urea, and (15)N enrichment in feces were used to calculate urea kinetics. For the low-CP diets, cows fed the high-RDP diet had a greater DM intake compared with those fed the low-RDP diet, but the opposite trend was observed for cows fed the high-CP diets. Dietary treatment had no effect on milk yield. Milk composition and milk component yields were largely unaffected by dietary treatment; however, on the low-CP diets, milk fat yield was greater for cows fed the low-RDP diet compared with those fed the high-RDP diet, but it was unaffected by RDP concentration on the high-CP diets. On the high-CP diets, milk urea nitrogen concentration was greater in cows fed the high-RDP diet compared with those fed the low-RDP diet, but it was unaffected by RDP concentration on the low-CP diets. Ruminal NH3-N concentration tended to

  11. Rumen metabolism of swamp buffaloes fed rice straw supplemented with cassava hay and urea.

    PubMed

    Ampapon, Thiwakorn; Wanapat, Metha; Kang, Sungchhang

    2016-04-01

    The objectives of this experiment were to investigate effects of cassava hay (CH) and urea (U) supplementation on feed intake, digestibility, rumen fermentation, and microbial protein synthesis of swamp buffaloes fed on rice straw. Four rumen-fistulated swamp buffaloes, 365 ± 15.0 kg, were randomly assigned according to a 4 × 4 Latin square design to receive four dietary treatments: T1 = CH 400 g/head/day + U 0 g/head/day, T2 = CH + U 30 g/head/day, T3 = CH + U 60 g/head/day, and T4 = CH + U 90 g/head/day, respectively. Results revealed that feed intake was not affected while nutrient digestibilities were increased (P < 0.05) with increasing U level supplementation especially at 90 g/head/day. Ruminal pH and temperature were not altered by urea supplementation, whereas ammonia nitrogen (NH3-N) and blood urea nitrogen were increased with urea supplement (P < 0.05). In addition, total volatile fatty acid and butyric acid were similar among treatments, while propionic acid (C3) was increased by level of urea supplement (P < 0.05), but acetic acid (C2) and C2/C3 ratio were significantly decreased (P < 0.05). On the other hand, protozoal population and methane production were decreased by CH and urea supplement, while bacterial population particularly those of proteolytic, cellulolytic, and amylolytic bacteria and efficiency of microbial nitrogen synthesis were linearly increased (P < 0.05). Based on this experiment, it suggested that supplementation of urea and cassava hay for buffaloes fed rice straw improved rumen ecology and increased fermentation end products and microbial protein synthesis while reducing protozoal populations and methane production. Urea supplements of 60-90 g/head/day when fed with cassava hay are recommended for swamp buffaloes consuming rice straw.

  12. Perturbation dynamics of the rumen microbiota in response to exogenous butyrate.

    PubMed

    Li, Robert W; Wu, Sitao; Baldwin, Ransom L; Li, Weizhong; Li, Congjun

    2012-01-01

    The capacity of the rumen microbiota to produce volatile fatty acids (VFAs) has important implications in animal well-being and production. We investigated temporal changes of the rumen microbiota in response to butyrate infusion using pyrosequencing of the 16S rRNA gene. Twenty one phyla were identified in the rumen microbiota of dairy cows. The rumen microbiota harbored 54.5±6.1 genera (mean ± SD) and 127.3±4.4 operational taxonomic units (OTUs), respectively. However, the core microbiome comprised of 26 genera and 82 OTUs. Butyrate infusion altered molar percentages of 3 major VFAs. Butyrate perturbation had a profound impact on the rumen microbial composition. A 72 h-infusion led to a significant change in the numbers of sequence reads derived from 4 phyla, including 2 most abundant phyla, Bacteroidetes and Firmicutes. As many as 19 genera and 43 OTUs were significantly impacted by butyrate infusion. Elevated butyrate levels in the rumen seemingly had a stimulating effect on butyrate-producing bacteria populations. The resilience of the rumen microbial ecosystem was evident as the abundance of the microorganisms returned to their pre-disturbed status after infusion withdrawal. Our findings provide insight into perturbation dynamics of the rumen microbial ecosystem and should guide efforts in formulating optimal uses of probiotic bacteria treating human diseases.

  13. Steers grazing blue grama rangeland throughout the growing season. II. Site and extent of digestion and microbial protein synthesis.

    PubMed

    Funk, M A; Galyean, M L; Branine, M E

    1987-11-01

    Effects of advancing forage maturity and drought-induced summer dormancy on site and extent of digestion and microbial protein synthesis in beef steers grazing native blue grama rangeland were evaluated in four sampling periods. Five steers (avg initial wt 227 kg) fitted with ruminal, duodenal and ileal cannulae and three steers cannulated at the esophagus freely grazed a 12-ha study pasture. Sampling periods lasted 11 d and started June 2, which was during the early growing season (EGS); June 22, during early summer dormancy (ESD); July 21, during late summer dormancy (LSD); and August 25, 1985, during the late growing season (LGS). Dietary N content was lower (P less than .05) in ESD and LSD than in EGS and LGS. Neutral detergent fiber (NDF) content was lower (P less than .05) in EGS than in other sampling periods. Ruminal organic matter (OM) digestion was lower (P less than .05) in ESD than in EGS, probably because of increased dietary NDF and lower N content. Ruminal OM digestion was greater (P less than .05) in LSD and LGS than in ESD because of increased fiber digestion. Neutral detergent fiber and acid detergent fiber (ADF) digestion occurring in the rumen was greater (P less than .05) in LSD and LGS than in EGS and ESD. Organic matter digestion in the small intestine and OM, NDF and ADF digestion in the hindgut were similar for all sampling periods. Over 90% of the fiber digestion occurred ruminally.(ABSTRACT TRUNCATED AT 250 WORDS)

  14. Manipulation of Rumen Microbial Fermentation by Polyphenol Rich Solvent Fractions from Papaya Leaf to Reduce Green-House Gas Methane and Biohydrogenation of C18 PUFA.

    PubMed

    Jafari, Saeid; Meng, Goh Yong; Rajion, Mohamed Ali; Jahromi, Mohammad Faseleh; Ebrahimi, Mahdi

    2016-06-01

    Different solvents (hexane, chloroform, ethyl acetate, butanol, and water) were used to identify the effect of papaya leaf (PL) fractions (PLFs) on ruminal biohydrogenation (BH) and ruminal methanogenesis in an in vitro study. PLFs at a concentration of 0 (control, CON) and 15 mg/250 mg dry matter (DM) were mixed with 30 mL of buffered rumen fluid and were incubated for 24 h. Methane (CH4) production (mL/250 mg DM) was the highest (P < 0.05) for CON (7.65) and lowest for the chloroform fraction (5.41) compared to those of other PLFs at 24 h of incubation. Acetate to propionate ratio was the lowest for PLFs compared to that of CON. Supplementation of the diet with PLFs significantly (P < 0.05) decreased the rate of BH of C18:1n-9 (oleic acid; OA), C18:2n-6 (linoleic acid; LA), and C18:3n-3 (α-linolenic acid; LNA) compared to that of CON after 24 h of incubation. Real time PCR indicated that total protozoa and total methanogen population in PLFs decreased (P < 0.05) compared to those of CON.

  15. Manipulation of Rumen Microbial Fermentation by Polyphenol Rich Solvent Fractions from Papaya Leaf to Reduce Green-House Gas Methane and Biohydrogenation of C18 PUFA.

    PubMed

    Jafari, Saeid; Meng, Goh Yong; Rajion, Mohamed Ali; Jahromi, Mohammad Faseleh; Ebrahimi, Mahdi

    2016-06-01

    Different solvents (hexane, chloroform, ethyl acetate, butanol, and water) were used to identify the effect of papaya leaf (PL) fractions (PLFs) on ruminal biohydrogenation (BH) and ruminal methanogenesis in an in vitro study. PLFs at a concentration of 0 (control, CON) and 15 mg/250 mg dry matter (DM) were mixed with 30 mL of buffered rumen fluid and were incubated for 24 h. Methane (CH4) production (mL/250 mg DM) was the highest (P < 0.05) for CON (7.65) and lowest for the chloroform fraction (5.41) compared to those of other PLFs at 24 h of incubation. Acetate to propionate ratio was the lowest for PLFs compared to that of CON. Supplementation of the diet with PLFs significantly (P < 0.05) decreased the rate of BH of C18:1n-9 (oleic acid; OA), C18:2n-6 (linoleic acid; LA), and C18:3n-3 (α-linolenic acid; LNA) compared to that of CON after 24 h of incubation. Real time PCR indicated that total protozoa and total methanogen population in PLFs decreased (P < 0.05) compared to those of CON. PMID:27192629

  16. Short communication: effects of level of rumen-degradable protein and corn distillers grains in corn silage-based diets on milk production and ruminal fermentation in lactating dairy cows.

    PubMed

    Zanton, G I; Heinrichs, A J; Jones, C M

    2013-07-01

    Two of the potential obstacles precluding inclusion of higher levels of dry distillers grains with solubles (DDGS) in corn-based dairy cow diets are the low levels of rumen-degradable protein (RDP) and the fatty acid content and composition of DDGS. Therefore, the objective of this experiment was to evaluate the production and rumen responses to dietary alterations in the level of RDP and DDGS for dairy cows fed a high corn silage diet. The experimental design was a replicated 4×4 Latin square with 21-d periods: 14 d of adaptation and 7 d of sampling; 16 uncannulated cows and 4 ruminally cannulated cows were blocked and assigned randomly to treatment sequences. Rations were provided as total mixed rations and were formulated to be high or low in RDP, with or without DDGS replacing soybean-based concentrates: high RDP, no DDGS (HRDP0); low RDP, no DDGS (LRDP0); low RDP, 10% DDGS; and low RDP, 20% DDGS. Body weight (696kg) and dry matter intake (26.6 kg/d) were not affected by treatment. Rumen ammonia concentration was greater for HRDP0 than LRDP0, but was unaffected by level of DDGS inclusion. The mean and minimum rumen pH and time pH was <5.5 were not different between diets. Milk production tended to be lower for cows fed HRDP0 than LRDP0 and tended to be linearly reduced as DDGS inclusion increased. Milk protein yield tended to be greater for cows fed LRDP0 than HRDP0, but was unaffected by DDGS level. Milk fat production, concentration, and fat-corrected milk were linearly reduced by increasing levels of DDGS. Based on these results, feeding DDGS at 20% of diet dry matter is not recommended for diets based on high corn silage.

  17. Effect of dietary nitrate level on enteric methane production, hydrogen emission, rumen fermentation, and nutrient digestibility in dairy cows.

    PubMed

    Olijhoek, D W; Hellwing, A L F; Brask, M; Weisbjerg, M R; Højberg, O; Larsen, M K; Dijkstra, J; Erlandsen, E J; Lund, P

    2016-08-01

    Nitrate may lower methane production in ruminants by competing with methanogenesis for available hydrogen in the rumen. This study evaluated the effect of 4 levels of dietary nitrate addition on enteric methane production, hydrogen emission, feed intake, rumen fermentation, nutrient digestibility, microbial protein synthesis, and blood methemoglobin. In a 4×4 Latin square design 4 lactating Danish Holstein dairy cows fitted with rumen, duodenal, and ileal cannulas were assigned to 4 calcium ammonium nitrate addition levels: control, low, medium, and high [0, 5.3, 13.6, and 21.1g of nitrate/kg of dry matter (DM), respectively]. Diets were made isonitrogenous by replacing urea. Cows were fed ad libitum and, after a 6-d period of gradual introduction of nitrate, adapted to the corn-silage-based total mixed ration (forage:concentrate ratio 50:50 on DM basis) for 16d before sampling. Digesta content from duodenum, ileum, and feces, and rumen liquid were collected, after which methane production and hydrogen emissions were measured in respiration chambers. Methane production [L/kg of dry matter intake (DMI)] linearly decreased with increasing nitrate concentrations compared with the control, corresponding to a reduction of 6, 13, and 23% for the low, medium, and high diets, respectively. Methane production was lowered with apparent efficiencies (measured methane reduction relative to potential methane reduction) of 82.3, 71.9, and 79.4% for the low, medium, and high diets, respectively. Addition of nitrate increased hydrogen emissions (L/kg of DMI) quadratically by a factor of 2.5, 3.4, and 3.0 (as L/kg of DMI) for the low, medium, and high diets, respectively, compared with the control. Blood methemoglobin levels and nitrate concentrations in milk and urine increased with increasing nitrate intake, but did not constitute a threat for animal health and human food safety. Microbial crude protein synthesis and efficiency were unaffected. Total volatile fatty acid

  18. Effect of dietary nitrate level on enteric methane production, hydrogen emission, rumen fermentation, and nutrient digestibility in dairy cows.

    PubMed

    Olijhoek, D W; Hellwing, A L F; Brask, M; Weisbjerg, M R; Højberg, O; Larsen, M K; Dijkstra, J; Erlandsen, E J; Lund, P

    2016-08-01

    Nitrate may lower methane production in ruminants by competing with methanogenesis for available hydrogen in the rumen. This study evaluated the effect of 4 levels of dietary nitrate addition on enteric methane production, hydrogen emission, feed intake, rumen fermentation, nutrient digestibility, microbial protein synthesis, and blood methemoglobin. In a 4×4 Latin square design 4 lactating Danish Holstein dairy cows fitted with rumen, duodenal, and ileal cannulas were assigned to 4 calcium ammonium nitrate addition levels: control, low, medium, and high [0, 5.3, 13.6, and 21.1g of nitrate/kg of dry matter (DM), respectively]. Diets were made isonitrogenous by replacing urea. Cows were fed ad libitum and, after a 6-d period of gradual introduction of nitrate, adapted to the corn-silage-based total mixed ration (forage:concentrate ratio 50:50 on DM basis) for 16d before sampling. Digesta content from duodenum, ileum, and feces, and rumen liquid were collected, after which methane production and hydrogen emissions were measured in respiration chambers. Methane production [L/kg of dry matter intake (DMI)] linearly decreased with increasing nitrate concentrations compared with the control, corresponding to a reduction of 6, 13, and 23% for the low, medium, and high diets, respectively. Methane production was lowered with apparent efficiencies (measured methane reduction relative to potential methane reduction) of 82.3, 71.9, and 79.4% for the low, medium, and high diets, respectively. Addition of nitrate increased hydrogen emissions (L/kg of DMI) quadratically by a factor of 2.5, 3.4, and 3.0 (as L/kg of DMI) for the low, medium, and high diets, respectively, compared with the control. Blood methemoglobin levels and nitrate concentrations in milk and urine increased with increasing nitrate intake, but did not constitute a threat for animal health and human food safety. Microbial crude protein synthesis and efficiency were unaffected. Total volatile fatty acid

  19. Milk from forage as affected by rumen degradable protein and corn grinding when feeding corn- and alfalfa silage-based diets.

    PubMed

    Charbonneau, E; Chouinard, P Y; Allard, G; Lapierre, H; Pellerin, D

    2007-02-01

    To increase the production of milk from forage (MF), a previous experiment with alfalfa silage showed the importance of a complementary combination of concentrates and forages offered. When corn silage is fed with alfalfa, increasing the rumen degradable protein (RDP) content in the diet should allow a better utilization of forage energy. To evaluate this hypothesis, 8 multiparous Holstein cows in early lactation were used in a replicated 4 x 4 Latin square design with 3-wk periods. Diets were fed as total mixed rations and were formulated to provide similar levels of net energy for lactation and crude protein but differing in RDP. Corn and alfalfa silages were used. Treatments were: 1) cracked corn-based concentrate providing low RDP [level recommended by the NRC (2001); RDP = 11.1% of dry matter (DM)]; 2) cracked corn-based concentrate providing medium RDP (RDP = 12.8% of DM); 3) cracked corn-based concentrate providing high RDP (RDP = 14.5% of DM); and 4) ground corn-based concentrate providing high RDP (RDP = 13.6% of DM). The first 3 treatments, using cracked corn, were compared on the basis of their RDP level. For these treatments, MF, calculated on a protein basis, decreased and the average of MF calculated on an energy basis and MF calculated on a protein basis tended to decrease as RDP increased. There was no difference for MF calculated on an energy basis between treatments. Increasing dietary RDP levels decreased the milk yield (from 32.8 to 30.7 kg/d) and milk protein yield (from 1,094 to 1,005 g/d) but not the milk fat yield. The milk urea N concentration increased as RDP increased. This suggests that there is no advantage of feeding RDP above the NRC recommendations when diets are based on corn and alfalfa silage. At high RDP levels (treatments 3 and 4), ground corn supported higher DM intake and yields of milk and protein than did cracked corn. Milk from forage, calculated on a protein basis, was higher and milk urea N decreased with ground corn

  20. Rumen management during aphagia.

    PubMed

    Shakespeare, A S

    2008-09-01

    Ruminants that for any reason are unable to eat enough to survive can be supported via rumen fistulation. To successfully accomplish this task, an understanding of rumen physiology is necessary. Some adaptation and modification of the normal physiological processes will be necessary because the extended time normally required to ingest food will, for obvious practical reasons, be reduced to a few minutes repeated once to three times a day. The physiology of significance to aphagic or dysphagic animals is discussed and relevant examples of clinical cases are used to illustrate practical applications.

  1. MannDB: A microbial annotation database for protein characterization

    SciTech Connect

    Zhou, C; Lam, M; Smith, J; Zemla, A; Dyer, M; Kuczmarski, T; Vitalis, E; Slezak, T

    2006-05-19

    MannDB was created to meet a need for rapid, comprehensive automated protein sequence analyses to support selection of proteins suitable as targets for driving the development of reagents for pathogen or protein toxin detection. Because a large number of open-source tools were needed, it was necessary to produce a software system to scale the computations for whole-proteome analysis. Thus, we built a fully automated system for executing software tools and for storage, integration, and display of automated protein sequence analysis and annotation data. MannDB is a relational database that organizes data resulting from fully automated, high-throughput protein-sequence analyses using open-source tools. Types of analyses provided include predictions of cleavage, chemical properties, classification, features, functional assignment, post-translational modifications, motifs, antigenicity, and secondary structure. Proteomes (lists of hypothetical and known proteins) are downloaded and parsed from Genbank and then inserted into MannDB, and annotations from SwissProt are downloaded when identifiers are found in the Genbank entry or when identical sequences are identified. Currently 36 open-source tools are run against MannDB protein sequences either on local systems or by means of batch submission to external servers. In addition, BLAST against protein entries in MvirDB, our database of microbial virulence factors, is performed. A web client browser enables viewing of computational results and downloaded annotations, and a query tool enables structured and free-text search capabilities. When available, links to external databases, including MvirDB, are provided. MannDB contains whole-proteome analyses for at least one representative organism from each category of biological threat organism listed by APHIS, CDC, HHS, NIAID, USDA, USFDA, and WHO. MannDB comprises a large number of genomes and comprehensive protein sequence analyses representing organisms listed as high

  2. Control of rumen methanogenesis.

    PubMed

    Van Nevel, C J; Demeyer, D I

    1996-09-01

    During the last decades, considerable research on methane production in the rumen and its inhibition has been carried out. Initially, as methane production represents a significant loss of gross energy in the feed (2-15%), the ultimate goal of such intervention in rumen fermentation was an increase in feed efficiency. A second reason favouring research on methane inhibition is its role in the global warming phenomenon and in the destruction of the ozone layer. In this review, the authors describe briefly several interventions for reducing methane emission by ruminants. The objective can be reached by intervention at the dietary level by ration manipulation (composition, feeding level) or by the use of additives or supplements. Examples of additives are polyhalogenated compounds, ionophores and other antibiotics. Supplementation of the ration with lipids also lowered methanogenesis. More biotechnological interventions, e.g., defaunation, probiotics and introduction of reductive acetogenesis in the rumen, are also mentioned. It can be concluded that drastic inhibition of methane production is not unequivocally successful as a result of several factors, such as: instantaneous inhibition often followed by restoration of methanogenesis due to adaptation of the microbes or degradation of the additive, toxicity for the host animal, negative effects on overall digestion and productive performance. Therefore, methanogenesis and its inhibition cannot be considered as a separate part of rumen fermentation and its consequences on the animal should be taken into account.

  3. Rumen microbiome from steers differing in feed efficiency.

    PubMed

    Myer, Phillip R; Smith, Timothy P L; Wells, James E; Kuehn, Larry A; Freetly, Harvey C

    2015-01-01

    The cattle rumen has a diverse microbial ecosystem that is essential for the host to digest plant material. Extremes in body weight (BW) gain in mice and humans have been associated with different intestinal microbial populations. The objective of this study was to characterize the microbiome of the cattle rumen among steers differing in feed efficiency. Two contemporary groups of steers (n=148 and n=197) were fed a ration (dry matter basis) of 57.35% dry-rolled corn, 30% wet distillers grain with solubles, 8% alfalfa hay, 4.25% supplement, and 0.4% urea for 63 days. Individual feed intake (FI) and BW gain were determined. Within contemporary group, the four steers within each Cartesian quadrant were sampled (n=16/group) from the bivariate distribution of average daily BW gain and average daily FI. Bacterial 16S rRNA gene amplicons were sequenced from the harvested bovine rumen fluid samples using next-generation sequencing technology. No significant changes in diversity or richness were indicated, and UniFrac principal coordinate analysis did not show any separation of microbial communities within the rumen. However, the abundances of relative microbial populations and operational taxonomic units did reveal significant differences with reference to feed efficiency groups. Bacteroidetes and Firmicutes were the dominant phyla in all ruminal groups, with significant population shifts in relevant ruminal taxa, including phyla Firmicutes and Lentisphaerae, as well as genera Succiniclasticum, Lactobacillus, Ruminococcus, and Prevotella. This study suggests the involvement of the rumen microbiome as a component influencing the efficiency of weight gain at the 16S level, which can be utilized to better understand variations in microbial ecology as well as host factors that will improve feed efficiency. PMID:26030887

  4. Rumen Microbiome from Steers Differing in Feed Efficiency

    PubMed Central

    2015-01-01

    The cattle rumen has a diverse microbial ecosystem that is essential for the host to digest plant material. Extremes in body weight (BW) gain in mice and humans have been associated with different intestinal microbial populations. The objective of this study was to characterize the microbiome of the cattle rumen among steers differing in feed efficiency. Two contemporary groups of steers (n=148 and n=197) were fed a ration (dry matter basis) of 57.35% dry-rolled corn, 30% wet distillers grain with solubles, 8% alfalfa hay, 4.25% supplement, and 0.4% urea for 63 days. Individual feed intake (FI) and BW gain were determined. Within contemporary group, the four steers within each Cartesian quadrant were sampled (n=16/group) from the bivariate distribution of average daily BW gain and average daily FI. Bacterial 16S rRNA gene amplicons were sequenced from the harvested bovine rumen fluid samples using next-generation sequencing technology. No significant changes in diversity or richness were indicated, and UniFrac principal coordinate analysis did not show any separation of microbial communities within the rumen. However, the abundances of relative microbial populations and operational taxonomic units did reveal significant differences with reference to feed efficiency groups. Bacteroidetes and Firmicutes were the dominant phyla in all ruminal groups, with significant population shifts in relevant ruminal taxa, including phyla Firmicutes and Lentisphaerae, as well as genera Succiniclasticum, Lactobacillus, Ruminococcus, and Prevotella. This study suggests the involvement of the rumen microbiome as a component influencing the efficiency of weight gain at the 16S level, which can be utilized to better understand variations in microbial ecology as well as host factors that will improve feed efficiency. PMID:26030887

  5. Effect of camelina oil or live yeasts (Saccharomyces cerevisiae) on ruminal methane production, rumen fermentation, and milk fatty acid composition in lactating cows fed grass silage diets.

    PubMed

    Bayat, A R; Kairenius, P; Stefański, T; Leskinen, H; Comtet-Marre, S; Forano, E; Chaucheyras-Durand, F; Shingfield, K J

    2015-05-01

    The potential of dietary supplements of 2 live yeast strains (Saccharomyces cerevisiae) or camelina oil to lower ruminal methane (CH4) and carbon dioxide (CO2) production and the associated effects on animal performance, rumen fermentation, rumen microbial populations, nutrient metabolism, and milk fatty acid (FA) composition of cows fed grass silage-based diets were examined. Four Finnish Ayrshire cows (53±7 d in milk) fitted with rumen cannula were used in a 4×4 Latin square with four 42-d periods. Cows received a basal total mixed ration (control treatment) with a 50:50 forage-to-concentrate ratio [on a dry matter (DM) basis] containing grass silage, the same basal total mixed ration supplemented with 1 of 2 live yeasts, A or B, administered directly in the rumen at 10(10) cfu/d (treatments A and B), or supplements of 60g of camelina oil/kg of diet DM that replaced concentrate ingredients in the basal total mixed ration (treatment CO). Relative to the control, treatments A and B had no effects on DM intake, rumen fermentation, ruminal gas production, or apparent total-tract nutrient digestibility. In contrast, treatment CO lowered DM intake and ruminal CH4 and CO2 production, responses associated with numerical nonsignificant decreases in total-tract organic matter digestibility, but no alterations in rumen fermentation characteristics or changes in the total numbers of rumen bacteria, methanogens, protozoa, and fungi. Compared with the control, treatment CO decreased the yields of milk, milk fat, lactose, and protein. Relative to treatment B, treatment CO improved nitrogen utilization due to a lower crude protein intake. Treatment A had no influence on milk FA composition, whereas treatment B increased cis-9 10:1 and decreased 11-cyclohexyl 11:0 and 24:0 concentrations. Treatment CO decreased milk fat 8:0 to 16:0 and total saturated FA, and increased 18:0, 18:1, 18:2, conjugated linoleic acid, 18:3n-3, and trans FA concentrations. Decreases in ruminal CH4

  6. The bacterial community composition of the bovine rumen detected using pyrosequencing of 16S rRNA genes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The rumen as a complex microbial ecosystem plays a critical role in sustainable agriculture. Rumen microorganisms perform important biochemical conversions, including the fermentation of plant fiber to small molecules such as short-chain fatty acids for meat and dairy production. In this study, we s...

  7. Selenite Reduction by Anaerobic Microbial Aggregates: Microbial Community Structure, and Proteins Associated to the Produced Selenium Spheres.

    PubMed

    Gonzalez-Gil, Graciela; Lens, Piet N L; Saikaly, Pascal E

    2016-01-01

    Certain types of anaerobic granular sludge, which consists of microbial aggregates, can reduce selenium oxyanions. To envisage strategies for removing those oxyanions from wastewater and recovering the produced elemental selenium (Se(0)), insights into the microbial community structure and synthesis of Se(0) within these microbial aggregates are required. High-throughput sequencing showed that Veillonellaceae (c.a. 20%) and Pseudomonadaceae (c.a.10%) were the most abundant microbial phylotypes in selenite reducing microbial aggregates. The majority of the Pseudomonadaceae sequences were affiliated to the genus Pseudomonas. A distinct outer layer (∼200 μm) of selenium deposits indicated that bioreduction occurred in the outer zone of the microbial aggregates. In that outer layer, SEM analysis showed abundant intracellular and extracellular Se(0) (nano)spheres, with some cells having high numbers of intracellular Se(0) spheres. Electron tomography showed that microbial cells can harbor a single large intracellular sphere that stretches the cell body. The Se(0) spheres produced by the microorganisms were capped with organic material. X-ray photoelectron spectroscopy (XPS) analysis of extracted Se(0) spheres, combined with a mathematical approach to analyzing XPS spectra from biological origin, indicated that proteins and lipids were components of the capping material associated to the Se(0) spheres. The most abundant proteins associated to the spheres were identified by proteomic analysis. Most of the proteins or peptide sequences capping the Se(0) spheres were identified as periplasmic outer membrane porins and as the cytoplasmic elongation factor Tu protein, suggesting an intracellular formation of the Se(0) spheres. In view of these and previous findings, a schematic model for the synthesis of Se(0) spheres by the microorganisms inhabiting the granular sludge is proposed. PMID:27199909

  8. Selenite Reduction by Anaerobic Microbial Aggregates: Microbial Community Structure, and Proteins Associated to the Produced Selenium Spheres

    PubMed Central

    Gonzalez-Gil, Graciela; Lens, Piet N. L.; Saikaly, Pascal E.

    2016-01-01

    Certain types of anaerobic granular sludge, which consists of microbial aggregates, can reduce selenium oxyanions. To envisage strategies for removing those oxyanions from wastewater and recovering the produced elemental selenium (Se0), insights into the microbial community structure and synthesis of Se0 within these microbial aggregates are required. High-throughput sequencing showed that Veillonellaceae (c.a. 20%) and Pseudomonadaceae (c.a.10%) were the most abundant microbial phylotypes in selenite reducing microbial aggregates. The majority of the Pseudomonadaceae sequences were affiliated to the genus Pseudomonas. A distinct outer layer (∼200 μm) of selenium deposits indicated that bioreduction occurred in the outer zone of the microbial aggregates. In that outer layer, SEM analysis showed abundant intracellular and extracellular Se0 (nano)spheres, with some cells having high numbers of intracellular Se0 spheres. Electron tomography showed that microbial cells can harbor a single large intracellular sphere that stretches the cell body. The Se0 spheres produced by the microorganisms were capped with organic material. X-ray photoelectron spectroscopy (XPS) analysis of extracted Se0 spheres, combined with a mathematical approach to analyzing XPS spectra from biological origin, indicated that proteins and lipids were components of the capping material associated to the Se0 spheres. The most abundant proteins associated to the spheres were identified by proteomic analysis. Most of the proteins or peptide sequences capping the Se0 spheres were identified as periplasmic outer membrane porins and as the cytoplasmic elongation factor Tu protein, suggesting an intracellular formation of the Se0 spheres. In view of these and previous findings, a schematic model for the synthesis of Se0 spheres by the microorganisms inhabiting the granular sludge is proposed. PMID:27199909

  9. Selenite Reduction by Anaerobic Microbial Aggregates: Microbial Community Structure, and Proteins Associated to the Produced Selenium Spheres.

    PubMed

    Gonzalez-Gil, Graciela; Lens, Piet N L; Saikaly, Pascal E

    2016-01-01

    Certain types of anaerobic granular sludge, which consists of microbial aggregates, can reduce selenium oxyanions. To envisage strategies for removing those oxyanions from wastewater and recovering the produced elemental selenium (Se(0)), insights into the microbial community structure and synthesis of Se(0) within these microbial aggregates are required. High-throughput sequencing showed that Veillonellaceae (c.a. 20%) and Pseudomonadaceae (c.a.10%) were the most abundant microbial phylotypes in selenite reducing microbial aggregates. The majority of the Pseudomonadaceae sequences were affiliated to the genus Pseudomonas. A distinct outer layer (∼200 μm) of selenium deposits indicated that bioreduction occurred in the outer zone of the microbial aggregates. In that outer layer, SEM analysis showed abundant intracellular and extracellular Se(0) (nano)spheres, with some cells having high numbers of intracellular Se(0) spheres. Electron tomography showed that microbial cells can harbor a single large intracellular sphere that stretches the cell body. The Se(0) spheres produced by the microorganisms were capped with organic material. X-ray photoelectron spectroscopy (XPS) analysis of extracted Se(0) spheres, combined with a mathematical approach to analyzing XPS spectra from biological origin, indicated that proteins and lipids were components of the capping material associated to the Se(0) spheres. The most abundant proteins associated to the spheres were identified by proteomic analysis. Most of the proteins or peptide sequences capping the Se(0) spheres were identified as periplasmic outer membrane porins and as the cytoplasmic elongation factor Tu protein, suggesting an intracellular formation of the Se(0) spheres. In view of these and previous findings, a schematic model for the synthesis of Se(0) spheres by the microorganisms inhabiting the granular sludge is proposed.

  10. Protein folding and conformational stress in microbial cells producing recombinant proteins: a host comparative overview

    PubMed Central

    Gasser, Brigitte; Saloheimo, Markku; Rinas, Ursula; Dragosits, Martin; Rodríguez-Carmona, Escarlata; Baumann, Kristin; Giuliani, Maria; Parrilli, Ermenegilda; Branduardi, Paola; Lang, Christine; Porro, Danilo; Ferrer, Pau; Tutino, Maria Luisa; Mattanovich, Diethard; Villaverde, Antonio

    2008-01-01

    Different species of microorganisms including yeasts, filamentous fungi and bacteria have been used in the past 25 years for the controlled production of foreign proteins of scientific, pharmacological or industrial interest. A major obstacle for protein production processes and a limit to overall success has been the abundance of misfolded polypeptides, which fail to reach their native conformation. The presence of misfolded or folding-reluctant protein species causes considerable stress in host cells. The characterization of such adverse conditions and the elicited cell responses have permitted to better understand the physiology and molecular biology of conformational stress. Therefore, microbial cell factories for recombinant protein production are depicted here as a source of knowledge that has considerably helped to picture the extremely rich landscape of in vivo protein folding, and the main cellular players of this complex process are described for the most important cell factories used for biotechnological purposes. PMID:18394160

  11. Comparative analysis of Escherichia coli O157 growth and protein-expression, in vitro and in vivo, in rumen fluid of cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cattle are the primary reservoirs for Escherichia coli O157 (O157), a Shiga toxin-producing E. coli, with potential for serious extraintestinal sequelae in humans. In a recent study (Kudva IT et al. BMC Microbiol. 2014; 14:48), we reported that when cultured in rumen fluid from dairy cattle on the ...

  12. [The course of rumen fermentation during alkalosis in cows].

    PubMed

    Zawadzki, W; Hejłasz, Z; Nicpoń, J

    1991-01-01

    The aim of the study was the investigation of rumen fermentation during alkalosis in cows. The study comprised some parameters of rumen fermentation, such as: pH, ammonia and volatile fatty acids (VFA) levels, also relationship between VFA, numbers of population of protozoa and bacteria, total production of gases in vitro particularly CO2 and CH4, amounts of lactic and total protein in rumen fluid and non-glucogenic/glucogenic ratio (NGGR) in the VFA mixture. On the basis of obtained results the amounts of fermented hexose, cell yield, ATP produced and hydrogen utilization were calculated. During alkalosis there was observed significant fall of VFA production, especially acetic and butyric acids, also lower production of gases, particularly CH4--probably as a result of selective reduction of methanogenic strain bacteria. The levelling of value of rumen pattern of fermentation occurred after the beginning of lactation probably as a result of metabolism products excretion together with milk. PMID:1842617

  13. Effect of quebracho-chestnut tannin extracts at 2 dietary crude protein levels on performance, rumen fermentation, and nitrogen partitioning in dairy cows.

    PubMed

    Aguerre, M J; Capozzolo, M C; Lencioni, P; Cabral, C; Wattiaux, M A

    2016-06-01

    Our objective was to determine the effects of a tannin mixture extract on lactating cow performance, rumen fermentation, and N partitioning, and whether responses were affected by dietary crude protein (CP). The experiment was conducted as a split-plot with 24 Holstein cows (mean ± standard deviation; 669±55kg of body weight; 87±36 d in milk; 8 ruminally cannulated) randomly assigned to a diet of [dry matter (DM) basis] 15.3 or 16.6% CP (whole plot) and 0, 0.45, 0.90, or 1.80% of a tannin mixture in three 4×4 Latin squares within each level of CP (sub-plot). Tannin extract mixture was from quebracho and chestnut trees (2:1 ratio). Dietary CP level did not influence responses to tannin supplementation. A linear decrease in DM intake (25.5 to 23.4kg/d) was found, as well as a linear increase in milk/DM intake (1.62 to 1.75) and a trend for a linear decrease in fat-and-protein-corrected milk (38.4 to 37.1kg/d) with increasing levels of tannin supplementation. In addition, there was a negative linear effect for milk urea N (14.0 to 12.9mg/dL), milk protein yield (1.20 to 1.15kg), and concentration (2.87 to 2.83%). Furthermore, the change in milk protein concentration tended to be quadratic, and predicted maximum was 2.89% for a tannin mixture fed at 0.47% of dietary DM. Tannin supplementation reduced ruminal NH3-N (11.3 to 8.8mg/dL), total branched-chain volatile fatty acid concentration (2.97 to 2.47mol/100mol), DM, organic matter, CP, and neutral detergent fiber digestibility. Dietary tannin had no effect on intake N (587±63g/d), milk N (175±32g/d), or N utilization efficiency (29.7±4.4%). However, feeding tannin extracts linearly increased fecal N excretion (214 to 256g/d), but reduced urinary N (213 to 177g/d) and urinary urea N (141 to 116g/d) excretion. Decreasing dietary CP did not influence milk production, but increased N utilization efficiency (milk N/N intake; 0.27 to 0.33), and decreased milk urea N (15.4 to 11.8mg/dL), ruminal NH3-N (11.0 to 9.3mg

  14. Isolation and characterization of a non-specific endoglucanase from a metagenomic library of goat rumen.

    PubMed

    Cheng, Jianbo; Huang, Shuai; Jiang, Haiqin; Zhang, Yunhai; Li, Lvmu; Wang, Juhua; Fan, Caiyun

    2016-01-01

    A cellulase gene (cel28a) was isolated from a rumen microbial metagenome library of goat rumen microorganisms, cloned into E. coli, and expressed in active form. The gene has a length of 1596 bp obtained using a genome walking Kit and encodes a protein of 509 amino acids with a calculated MW of 55 kDa. The deduced amino acid sequence was homologous with cellulases belonging to the glycosyl hydrolase family 5 (GH5). The expressed protein showed activity toward carboxymethylcellulose (CMC) and xylan, suggesting non-specific endoglucanase activity. The optimal conditions for endoglucanase and xylanase activities were 50 °C and pH 5.0. The metal ions (Ca(2+), Fe(2+), Mn(2+) and Co(2+)) stimulated the cellulase activity of cel28a, while the other metal ions and chemicals (Ni(2+), Mg(2+), Zn(2+), Cu(2+), SDS and EDTA) inhibited the cellulase activity. Further examination of substrate preference showed a higher activity with CMC, oat spelt xylan and birchwood xylan than with filter paper and microcrystalline cellulose, again suggesting that the protein was an endoglucanase with xylanase activity.

  15. Isolation and characterization of a non-specific endoglucanase from a metagenomic library of goat rumen.

    PubMed

    Cheng, Jianbo; Huang, Shuai; Jiang, Haiqin; Zhang, Yunhai; Li, Lvmu; Wang, Juhua; Fan, Caiyun

    2016-01-01

    A cellulase gene (cel28a) was isolated from a rumen microbial metagenome library of goat rumen microorganisms, cloned into E. coli, and expressed in active form. The gene has a length of 1596 bp obtained using a genome walking Kit and encodes a protein of 509 amino acids with a calculated MW of 55 kDa. The deduced amino acid sequence was homologous with cellulases belonging to the glycosyl hydrolase family 5 (GH5). The expressed protein showed activity toward carboxymethylcellulose (CMC) and xylan, suggesting non-specific endoglucanase activity. The optimal conditions for endoglucanase and xylanase activities were 50 °C and pH 5.0. The metal ions (Ca(2+), Fe(2+), Mn(2+) and Co(2+)) stimulated the cellulase activity of cel28a, while the other metal ions and chemicals (Ni(2+), Mg(2+), Zn(2+), Cu(2+), SDS and EDTA) inhibited the cellulase activity. Further examination of substrate preference showed a higher activity with CMC, oat spelt xylan and birchwood xylan than with filter paper and microcrystalline cellulose, again suggesting that the protein was an endoglucanase with xylanase activity. PMID:26712627

  16. Different forms of rumen dystonia in dairy cows.

    PubMed

    Sederevicius, A; Kantautaité, J

    1993-01-01

    Four naturally occurring main forms of rumen dystonia of alimentary origin were observed in cattle: acidotic conditions; microflora inactivity; acidosis, and alkalosis. The following disturbances in the metabolism of carbohydrates, protein and energy were observed: 1. Acidotic conditions in rumen--ruminal pH, total and protein nitrogen decreased, whereas the total amount of VFA and the reduction activity of bacteria increased. 2. Inactivity of rumen microflora--significant changes of pH of rumen fluid were not observed, but the total number and activity of infusoria decreased. Fermentation of glucose, digestibility of cellulose, and reduction activity of bacteria decreased, whereas the amount of non-protein nitrogen increased. 3. Acidosis--ruminal pH, reduction activity of bacteria, and total number of VFA decreased. The percentage ratio between VFA changed--acetic acid concentration decreased, the concentration of valeric and caproic acids increased. The amount of total and non-protein nitrogen increased. 4. Alkalosis--ruminal pH increased, reduction activity of bacteria, fermentation of glucose, and concentration of VFA decreased. The amount of total and non-protein nitrogen increased. This investigation of different forms of rumen dystonia of alimentary origin is believed to be useful for the development of more effective treating methods and measures.

  17. Metaproteomics Reveals Functional Shifts in Microbial and Human Proteins During Infant Gut Colonization Case

    SciTech Connect

    Young, Jacque C.; Pan, Chongle; Adams, Rachel M.; Brooks, Brandon; Banfield, Jillian F.; Morowitz, Michael J.; Robert L. Hettich

    2015-01-01

    The microbial colonization of the human gastrointestinal tract plays an important role in establishing health and homeostasis. However, the time-dependent functional signatures of microbial and human proteins during early colonization of the gut have yet to be determined. Thus, we employed shotgun proteomics to simultaneously monitor microbial and human proteins in fecal samples from a preterm infant during the first month of life. Microbial community complexity and functions increased over time, with compositional changes that were consistent with previous metagenomic and rRNA gene data indicating three distinct colonization phases. Overall microbial community functions were established relatively early in development and remained stable. Human proteins detected included those responsible for epithelial barrier function and antimicrobial activity. Some neutrophil-derived proteins increased in abundance early in the study period, suggesting activation of the innate immune system. Moreover, abundances of cytoskeletal and mucin proteins increased later in the time course, suggestive of subsequent adjustment to the increased microbial load. Our study provides the first snapshot of coordinated human and microbial protein expression in the infant gut during early development.

  18. Metaproteomics Reveals Functional Shifts in Microbial and Human Proteins During Infant Gut Colonization Case

    DOE PAGESBeta

    Young, Jacque C.; Pan, Chongle; Adams, Rachel M.; Brooks, Brandon; Banfield, Jillian F.; Morowitz, Michael J.; Robert L. Hettich

    2015-01-01

    The microbial colonization of the human gastrointestinal tract plays an important role in establishing health and homeostasis. However, the time-dependent functional signatures of microbial and human proteins during early colonization of the gut have yet to be determined. Thus, we employed shotgun proteomics to simultaneously monitor microbial and human proteins in fecal samples from a preterm infant during the first month of life. Microbial community complexity and functions increased over time, with compositional changes that were consistent with previous metagenomic and rRNA gene data indicating three distinct colonization phases. Overall microbial community functions were established relatively early in development andmore » remained stable. Human proteins detected included those responsible for epithelial barrier function and antimicrobial activity. Some neutrophil-derived proteins increased in abundance early in the study period, suggesting activation of the innate immune system. Moreover, abundances of cytoskeletal and mucin proteins increased later in the time course, suggestive of subsequent adjustment to the increased microbial load. Our study provides the first snapshot of coordinated human and microbial protein expression in the infant gut during early development.« less

  19. Screening for the effects of natural plant extracts at different pH on in vitro rumen microbial fermentation of a high-concentrate diet for beef cattle.

    PubMed

    Cardozo, P W; Calsamiglia, S; Ferret, A; Kamel, C

    2005-11-01

    Six natural plant extracts and three secondary plant metabolites were tested at five doses (0, 0.3, 3, 30, and 300 mg/L) and two different pH (7.0 and 5.5) in a duplicate 9 x 5 x 2 factorial arrangement of treatments to determine their effects on in vitro microbial fermentation using ruminal fluid from heifers fed a high-concentrate finishing diet. Treatments were extracts of garlic (GAR), cinnamon (CIN), yucca (YUC), anise (ANI), oregano (ORE), and capsicum (CAP) and pure cinnamaldehyde (CDH), anethole (ATL), and eugenol (EUG). Each treatment was tested in triplicate and in two periods. Fifty milliliters of a 1:1 ruminal fluid-to-buffer solution were introduced into polypropylene tubes supplied with 0.5 g of DM of a 10:90 forage:concentrate diet (15.4% CP, 16.0% NDF; DM basis) and incubated for 24 h at 39 degrees C. Samples were collected for ammonia N and VFA concentrations. The decrease in pH from 7.0 to 5.5 resulted in lower (P < 0.05) total VFA, ammonia N, branched-chain VFA concentration, acetate proportion, and acetate:propionate, and in a higher (P < 0.05) propionate proportion. The interaction between pH and doses was significant for all measurements, except for ATL and CDH for butyrate, ATL and EUG for acetate:propionate ratio, and ORE for ammonia N concentration. The high dose of all plant extracts decreased (P < 0.05) total VFA concentrations. When pH was 7.0, ATL, GAR, CAP, and CDH decreased (P < 0.05) total VFA concentration, and ANI, ORE, CIN, CAP, and CDH increased (P < 0.05) the acetate:propionate. The CIN, GAR, CAP, CDH, ORE, and YUC decreased (P < 0.05), and EUG, ANI, and ATL increased (P < 0.05) ammonia N concentration. The effects of plant extracts on the fermentation profile when pH was 7.0 were not favorable for beef production. In contrast, when pH was 5.5, total VFA concentration did not change (ATL, ANI, ORE, and CIN) or increased (P < 0.05) (EUG, GAR, CAP, CDH, and YUC), and the acetate:propionate (ORE, GAR, CAP, CDH, and YUC) decreased

  20. Effects of Rumen-Mate on lactational performance of Holsteins fed a high grain diet.

    PubMed

    Solorzano, L C; Armentano, L E; Emery, R S; Schricker, B R

    1989-07-01

    Three Latin-square trials were conducted to determine the effects of supplementing Rumen-Mate, a commercial buffer containing KCl, NaCl, and Mg and Na carbonates, on lactation performance of Holsteins. Cows were fed a basal ration of 40% corn silage and 60% concentrate in Trials 1 and 2, and 40% corn silage, 55% concentrate, and 5% alfalfa hay in Trial 3 (DM basis). In Trial 1, treatments were: basal diet, or basal diet supplemented with either 1% NaHCO3, or 1, 3, or 4.4% Rumen-Mate. Increasing dietary Rumen-Mate resulted in a linear increase in milk fat production and concentration with no difference between 1% Rumen-Mate and 1% bicarbonate. There was a significant linear decrease in milk protein concentration, but not production, with increasing concentrations of Rumen-Mate. In Trial 2 treatments were: basal diet, or basal diet supplemented with either .8% NaHCO3, 2.6% Rumen-Mate, .5% MgO, .8% NaHCO3 plus .5% MgO, or 1.8% Rumen-Mate plus .8% NaHCO3. Organic matter and CP intakes and milk protein yield and concentration were decreased by Rumen-Mate with a nonsignificant increase in milk fat concentration. Data from Trials 1 and 2 were combined with data from Trial 3, which compared basal diet, 1% bicarbonate, and 3% Rumen-Mate. The combined data showed a larger increase over basal diet in milk fat yield and concentration for 2.6 to 3% Rumen-Mate vs. .8 to 1% bicarbonate. Rumen-Mate did not decrease DM intake or protein yield relative to basal diet but did decrease protein yield 34 g/d compared with that of bicarbonate.

  1. Production of tyrosine and other aromatic compounds from phenylalanine by rumen microorganisms.

    PubMed

    Khan, R I; Onodera, R; Amin, M R; Mohammed, N

    1999-01-01

    Rumen contents from three fistulated Japanese native goats fed Lucerne hay cubes (Medicago sativa) and concentrate mixture were collected to prepare the suspensions of mixed rumen bacteria (B), mixed protozoa (P) and a combination of the two (BP). Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h with or without 1 mM of L-phenylalanine (Phe). Phe, tyrosine (Tyr) and other related compounds in both supernatant and microbial hydrolysates of the incubations were analyzed by HPLC. Tyr can be produced from Phe not only by rumen bacteria but also by rumen protozoa. The production of Tyr during 12 h incubation in B (183.6 mumol/g MN) was 4.3 times higher than that in P. One of the intermediate products between Phe and Tyr seems to be p-hydroxyphenylacetic acid. The rate of the net degradation of Phe incubation in B (76.0 mumol/g MN/h) was 2.4 times higher than in P. In the case of all rumen microorganisms, degraded Phe was mainly (> 53%) converted into phenylacetic acid. The production of benzoic acid was higher in P than in B suspensions. Small amount of phenylpyruvic acid was produced from Phe by both rumen bacteria and protozoa, but phenylpropionic acid and phenyllactic acid were produced only by rumen bacteria.

  2. [The effect of monensin on the fermentation of feed with different proportions of hay and concentrate in an artificial rumen (rumen simulation technic)].

    PubMed

    Jalc, D; Baran, M; Vendrák, T; Siroka, P

    1991-01-01

    An experiment was conducted with rumen pouch (RUSITEC--Rumen Simulation Technique). In four fermentation vessels (V), percent proportions of hay and barley were as follows: V1--40:60, V2 - 60:40, V3--80:20 and V4--100:0. Every day 5 mg of monensin dissolved in 1 ml 96% ethanol were added to each fermentation vessel. All diets were isonitrogenous, and after an addition of urea the crude protein (CP) content made 13% in each diet. The experiment lasted 12 days: so called steady state period took the first six days when the fermentation conditions were stabilized. Monensin reduced dry matter digestibility, production of total volatile fatty acids, acetic acid, n-butyric and isovaleric acids and acetate: propionate proportion, and it increased the production of propionic and n-valeric acids. The production of methane and CO2 decreased. The higher proportion of hay in diets decreased dry matter digestibility, digestibility of detergent fibre, total and individual volatile fatty acids, CO2, methane energy yield of volatile fatty acids (E), glucose utilization, production of adenosine triphosphate and production of fermented hexoses. The production, utilization and recovery of metabolic hydrogen also decreased. The effectiveness of microbial matter synthesis (YATP = 11.3) was highest during the fermentation of feed containing 60% hay and 40% barley. PMID:1926680

  3. Rumen modulatory effect of thyme, clove and peppermint oils in vitro using buffalo rumen liquor

    PubMed Central

    Roy, Debashis; Tomar, S. K.; Kumar, Vinod

    2015-01-01

    Aim: The present study was conducted to examine the rumen modulatory effect of thyme, clove and peppermint oils on rumen fermentation pattern in vitro using roughage based diet. Materials and Methods: Thyme, clove and peppermint oils were tested at concentration of 0, 30, 300 and 600 mg/l (ppm) of total culture fluid using in vitro gas production technique in wheat straw based diet (concentrate: Wheat straw 50:50). Different in vitro parameters e.g., total gas production, methane production, nutrient degradability, volatile fatty acid (VFA) production and ammonia nitrogen concentration were studied using buffalo rumen liquor. Results: Thyme oil at higher dose level (600 ppm) reduced (p<0.05) total gas production, feed degradability and ammonia nitrogen (NH3-N) concentration whereas total VFA concentration was significantly lower (p>0.05) in 300 and 600 ppm dose levels. 600 ppm dose level of clove oil reduced (p<0.05) total gas production, feed degradability, total VFA and acetate to propionate ratio. Methane production was significantly reduced (p<0.05) in 300 and 600 ppm dose levels of clove and peppermint oil. Conclusion: Right combination of these essential oils may prove to enhance performance of animals by reducing methane production and inhibiting protein degradation in rumen. PMID:27047073

  4. Effects of ethanol and heat treatments of soybean meal and infusion of sodium chloride into the rumen on ruminal degradation and escape of soluble and total soybean meal protein in steers.

    PubMed

    Lynch, G L; Berger, L L; Merchen, N R; Fahey, G C; Baker, E C

    1987-12-01

    Soybean meal (SBM) treated with 70% ethanol at 80 C (ET), nontreated SBM (NT) or a ureacasein-corn mix (UC) was fed to steers fitted with ruminal and duodenal cannulae to study ruminal N metabolism. Sodium chloride (NaCl) was ruminally infused at 0 or 500 g/d. Nitrogen supplements provided approximately 70% of total dietary N. Experimental design was a 6 X 6 Latin square with a 3 X 2 factorial arrangement of treatments. Total duodenal N flows and non-ammonia, non-bacterial-N (NANB-N) flows were higher (P less than .05) when steers were fed SBM treatments compared with UC, and higher (P less than .05) when steers were fed ET compared with NT. Percentage of SBM-N escaping ruminal degradation was greater (P less than .05) when steers were fed ET compared with NT, and greater (P less than .05) when NaCl was infused into the rumen. Duodenal flows of total, indispensible and dispensible amino acids were increased (P less than .05) when steers were fed SBM treatments compared with UC, and greater (P less than .05) when steers were fed ET compared with NT. No differences in soluble N flows at the omasum were observed due to treatment. Bacterial protein comprised the majority of the N leaving the rumen. Both ruminal NaCl infusion and ethanol and heat treatment of SBM increased ruminal SBM-N escape.

  5. Effect of the type of silage on milk yield, intake and rumen metabolism of dairy cows grazing swards with low herbage mass.

    PubMed

    Ruiz-Albarrán, Miguel; Balocchi, Oscar A; Noro, Mirela; Wittwer, Fernando; Pulido, Rubén G

    2016-07-01

    The aim of this study was to evaluate the effect of herbage allowance (HA) and type of silage supplemented (TS) on milk yield, dry matter intake (DMI) and metabolism of dairy cows in early lactation. Thirty-six Holstein-Friesian dairy cows were allocated to four treatments derived from an arrangement of two HA (LHA = 17 or HHA = 25 kg of DM/cow/day) and two TS (grass (GS) or maize (MS)). Herbage allowance had no effect on DMI or milk yield. Rumen pH and NH3 -N concentration were not affected by HA. The efficiency of microbial protein synthesis in the rumen (microbial protein (MP)) was affected by HA with 21.5 and 23.9 g microbial nitrogen per kg ruminal digestible organic matter for LHA and HHA, respectively (P < 0.05). Supplementation with MS showed higher values of milk yield by 2.4 kg/cow/day (P < 0.001), milk protein content by 0.10 % (P < 0.023) and herbage DMI by 2.2 kg/cow/day, and showed lower values for milk urea compared to GS (P < 0.001). The former results suggest that TS had a greater effect on milk yield, total feed intake and energy intake than increase in herbage allowance; however, increase in HA had greater effects on MP than TS.

  6. Effects of Eucalyptus Crude Oils Supplementation on Rumen Fermentation, Microorganism and Nutrient Digestibility in Swamp Buffaloes

    PubMed Central

    Thao, N. T.; Wanapat, M.; Cherdthong, A.; Kang, S.

    2014-01-01

    not affect nitrogen utilization. Both allantoin excretion and absorption and microbial nitrogen supply were increased by UTRS whereas efficiency of microbial protein synthesis was similar in all treatments. Findings of present study suggested that EuO could be used as a feed additive to modify the rumen fermentation in reducing methane production both in RS and UTRS. Feeding UTRS could improve feed intake and efficiency of rumen fermentation in swamp buffaloes. However, more research is warranted to determine the effect of EuO supplementation in production animals. PMID:25049925

  7. Manipulating rumen microbiome and fermentation through interventions during early life: a review.

    PubMed

    Yáñez-Ruiz, David R; Abecia, Leticia; Newbold, Charles J

    2015-01-01

    The nutritional manipulations of the rumen microbiome to enhance productivity and health are rather limited by the resilience of the ecosystem once established in the mature rumen. Based on recent studies, it has been suggested that the microbial colonization that occurs soon after birth opens a possibility of manipulation with potential to produce lasting effects into adult life. This paper presents the state-of-the-art in relation to early life nutritional interventions by addressing three areas: the development of the rumen as an organ in regards to the nutrition of the new-born, the main factors that determine the microbial population that first colonizes and establishes in the rumen, and the key immunity players that contribute to shaping the commensal microbiota in the early stage of life to understand host-microbiome specificity. The development of the rumen epithelium and muscularization are differently affected by the nature of the diet and special care should be taken with regards to transition from liquid (milk) to solid feed. The rumen is quickly colonized by all type of microorganisms straight after birth and the colonization pattern may be influenced by several factors such as presence/absence of adult animals, the first solid diet provided, and the inclusion of compounds that prevent/facilitate the establishment of some microorganisms or the direct inoculation of specific strains. The results presented show how early life events may be related to the microbial community structure and/or the rumen activity in the animals post-weaning. This would create differences in adaptive capacity due to different early life experiences and leads to the idea of microbial programming. However, many elements need to be further studied such as: the most sensitive window of time for interventions, the best means to test long term effectiveness, the role of key microbial groups and host-immune regulations.

  8. Manipulating rumen microbiome and fermentation through interventions during early life: a review.

    PubMed

    Yáñez-Ruiz, David R; Abecia, Leticia; Newbold, Charles J

    2015-01-01

    The nutritional manipulations of the rumen microbiome to enhance productivity and health are rather limited by the resilience of the ecosystem once established in the mature rumen. Based on recent studies, it has been suggested that the microbial colonization that occurs soon after birth opens a possibility of manipulation with potential to produce lasting effects into adult life. This paper presents the state-of-the-art in relation to early life nutritional interventions by addressing three areas: the development of the rumen as an organ in regards to the nutrition of the new-born, the main factors that determine the microbial population that first colonizes and establishes in the rumen, and the key immunity players that contribute to shaping the commensal microbiota in the early stage of life to understand host-microbiome specificity. The development of the rumen epithelium and muscularization are differently affected by the nature of the diet and special care should be taken with regards to transition from liquid (milk) to solid feed. The rumen is quickly colonized by all type of microorganisms straight after birth and the colonization pattern may be influenced by several factors such as presence/absence of adult animals, the first solid diet provided, and the inclusion of compounds that prevent/facilitate the establishment of some microorganisms or the direct inoculation of specific strains. The results presented show how early life events may be related to the microbial community structure and/or the rumen activity in the animals post-weaning. This would create differences in adaptive capacity due to different early life experiences and leads to the idea of microbial programming. However, many elements need to be further studied such as: the most sensitive window of time for interventions, the best means to test long term effectiveness, the role of key microbial groups and host-immune regulations. PMID:26528276

  9. Manipulating rumen microbiome and fermentation through interventions during early life: a review

    PubMed Central

    Yáñez-Ruiz, David R.; Abecia, Leticia; Newbold, Charles J.

    2015-01-01

    The nutritional manipulations of the rumen microbiome to enhance productivity and health are rather limited by the resilience of the ecosystem once established in the mature rumen. Based on recent studies, it has been suggested that the microbial colonization that occurs soon after birth opens a possibility of manipulation with potential to produce lasting effects into adult life. This paper presents the state-of-the-art in relation to early life nutritional interventions by addressing three areas: the development of the rumen as an organ in regards to the nutrition of the new-born, the main factors that determine the microbial population that first colonizes and establishes in the rumen, and the key immunity players that contribute to shaping the commensal microbiota in the early stage of life to understand host-microbiome specificity. The development of the rumen epithelium and muscularization are differently affected by the nature of the diet and special care should be taken with regards to transition from liquid (milk) to solid feed. The rumen is quickly colonized by all type of microorganisms straight after birth and the colonization pattern may be influenced by several factors such as presence/absence of adult animals, the first solid diet provided, and the inclusion of compounds that prevent/facilitate the establishment of some microorganisms or the direct inoculation of specific strains. The results presented show how early life events may be related to the microbial community structure and/or the rumen activity in the animals post-weaning. This would create differences in adaptive capacity due to different early life experiences and leads to the idea of microbial programming. However, many elements need to be further studied such as: the most sensitive window of time for interventions, the best means to test long term effectiveness, the role of key microbial groups and host-immune regulations. PMID:26528276

  10. The effect of rumen ciliates on chitinolytic activity, chitin content and the number of fungal zoospores in the rumen fluid of sheep.

    PubMed

    Miltko, Renata; Bełżecki, Grzegorz; Herman, Andrzej; Kowalik, Barbara; Skomiał, Jacek

    2016-12-01

    The objective of this study was to investigate the effect of selected protozoa on the degradation and concentration of chitin and the numbers of fungal zoospores in the rumen fluid of sheep. Three adult ewes were fed a hay-concentrate diet, defaunated, then monofaunated with Entodinium caudatum or Diploplastron affine alone and refaunated with natural rumen fauna. The average density of the protozoa population varied from 6.1 · 10(4) (D. affine) to 42.2 · 10(4) cells/ml rumen fluid (natural rumen fauna). The inoculation of protozoa in the rumen of defaunated sheep increased the total activity of chitinolytic enzymes from 2.9 to 3.6 μmol N-acetylglucosamine/g dry matter (DM) of rumen fluid per min, the chitin concentration from 6.3 to 7.2 mg/g DM of rumen fluid and the number of fungal zoospores from 8.1 to 10.9 · 10(5) cells/ml rumen fluid. All examined indices showed diurnal variations. Ciliate population density was highest immediately prior to feeding and lowest at 4 h thereafter. The opposite effects were observed for the numbers of fungal zoospores, the chitin concentration and chitinolytic activity. Furthermore, it was found that chitin from zoospores may account for up to 95% of total microbial chitin in the rumen fluid of sheep. In summary, the examined ciliate species showed the ability of chitin degradation as well as a positive influence on the development of the ruminal fungal population. PMID:27501267

  11. FERMENTATION CAPACITY AS A MEASURE OF NET GROWTH OF RUMEN MICROORGANISMS.

    PubMed

    EL-SHAZLY, K; HUNGATE, R E

    1965-01-01

    A simple technique for measuring the rate of fermentation of rumen microorganisms is described. It allows quick preparation and handling of the rumen sample immediately after collection. The average rate of fermentation of rumen samples collected from a lactating cow fed on alfalfa hay and concentrate in the ratio of 2:1 was very similar to the rate obtained by other methods. On the assumption that when substrate is in excess, the fermentation rate is proportional to the total microbial cells, the method was used to estimate the net growth of rumen microorganisms. The maximal fermentation rate of subsamples, taken at the beginning and after 1 hr of incubation of a sample, was measured. The results indicate a net average growth of 8% per hr, or 192% per day, in approximate agreement with rumen turnover time. The highest net growth does not necessarily coincide with the highest gas-production rate in the rumen, in part because the bicarbonate concentration in the rumen contents varies. In a cow fed on hay and concentrate, the net growth was lowest before feeding and immediately after feeding.

  12. High-throughput Methods Redefine the Rumen Microbiome and Its Relationship with Nutrition and Metabolism.

    PubMed

    McCann, Joshua C; Wickersham, Tryon A; Loor, Juan J

    2014-01-01

    Diversity in the forestomach microbiome is one of the key features of ruminant animals. The diverse microbial community adapts to a wide array of dietary feedstuffs and management strategies. Understanding rumen microbiome composition, adaptation, and function has global implications ranging from climatology to applied animal production. Classical knowledge of rumen microbiology was based on anaerobic, culture-dependent methods. Next-generation sequencing and other molecular techniques have uncovered novel features of the rumen microbiome. For instance, pyrosequencing of the 16S ribosomal RNA gene has revealed the taxonomic identity of bacteria and archaea to the genus level, and when complemented with barcoding adds multiple samples to a single run. Whole genome shotgun sequencing generates true metagenomic sequences to predict the functional capability of a microbiome, and can also be used to construct genomes of isolated organisms. Integration of high-throughput data describing the rumen microbiome with classic fermentation and animal performance parameters has produced meaningful advances and opened additional areas for study. In this review, we highlight recent studies of the rumen microbiome in the context of cattle production focusing on nutrition, rumen development, animal efficiency, and microbial function. PMID:24940050

  13. High-throughput Methods Redefine the Rumen Microbiome and Its Relationship with Nutrition and Metabolism

    PubMed Central

    McCann, Joshua C.; Wickersham, Tryon A.; Loor, Juan J.

    2014-01-01

    Diversity in the forestomach microbiome is one of the key features of ruminant animals. The diverse microbial community adapts to a wide array of dietary feedstuffs and management strategies. Understanding rumen microbiome composition, adaptation, and function has global implications ranging from climatology to applied animal production. Classical knowledge of rumen microbiology was based on anaerobic, culture-dependent methods. Next-generation sequencing and other molecular techniques have uncovered novel features of the rumen microbiome. For instance, pyrosequencing of the 16S ribosomal RNA gene has revealed the taxonomic identity of bacteria and archaea to the genus level, and when complemented with barcoding adds multiple samples to a single run. Whole genome shotgun sequencing generates true metagenomic sequences to predict the functional capability of a microbiome, and can also be used to construct genomes of isolated organisms. Integration of high-throughput data describing the rumen microbiome with classic fermentation and animal performance parameters has produced meaningful advances and opened additional areas for study. In this review, we highlight recent studies of the rumen microbiome in the context of cattle production focusing on nutrition, rumen development, animal efficiency, and microbial function. PMID:24940050

  14. Effects of increasing levels of grain supplementation on rumen environment and lactation performance of dairy cows grazing grass-legume pasture.

    PubMed

    Reis, R B; Combs, D K

    2000-12-01

    The impact of supplemental energy on nutrient utilization, fiber digestion, rumen fermentation, and lactation performance was evaluated in dairy cows grazing pastures composed of brome, orchardgrass, red clover, and alfalfa. Three amounts [0, 5, and 10 kg dry matter (DM)/d] of ground dry shelled corn-based concentrate were supplemented to nine rumen cannulated Holstein cows in a 3 x 3 Latin square replicated three times. Cows were on average 84+/-13 d in milk and producing 41.6+/-5.9 kg of milk/d at the beginning of the study. An increase in amounts of concentrate in the diets was associated with an increase in milk production, solids-corrected milk, and concentrations of milk protein and SNF. Milk fat percentage and milk urea nitrogen concentration decreased linearly with supplementation. Milk production and protein percentage were 21.8, 26.8, and 30.4 kg/d, and 2.85, 2.95, and 3.05% for the increasing levels of concentrate, respectively. Intake and digestibility of DM and organic matter (OM) increased as grain supplementation increased. Ruminal pH and total volatile fatty acid concentration (VFA) were not affected by supplementation or the amount of concentrate. Ruminal ammonia concentration was reduced by supplementation, presumably due to a decrease in N intake and greater use of ammonia-N for rumen microbial protein synthesis. Rumen fermentation varied throughout the day, with lower mean pH and higher VFA concentrations at night. Supplementation increased total OM intake, decreased forage OM intake, and increased the proportion of OM that was digested in the intestines. Total DM intake by grazing dairy cows can be increased using ground dry shelled corn-based concentrate without causing negative effects on forage digestion.

  15. Fluid absorption from the rumen during rehydration in sheep.

    PubMed

    Dahlborn, K; Holtenius, K

    1990-01-01

    Water movements across the rumen wall were studied during rehydration in four fistulated sheep. The animals were dehydrated for 48 h which increased total plasma protein, plasma osmolality, plasma Na+, arginine vasopressin and plasma renin activity. Two series of experiments were performed: Expt I with no food available during rehydration, and Expt II where the animals were fed hay. On the rehydration day, a fluid marker (cobalt-EDTA) was administered into the rumen. To avoid water outflow from the rumen a stopper was inserted into the reticulo-omasal orifice. When the animals were provided with water they immediately drank 9 l. The water offered contained the same marker concentration (Co2+) as in the rumen liquid. In Expt I, Co2+ concentration increased after drinking, and remained elevated until the stopper was removed. The highest value was obtained after 20 min, and this corresponded to at least a 11 water absorption in individual animals. In Expt II, the sheep immediately started to eat following drinking and the inflow of saliva caused a dilution of the marker. Plasma osmolality and Na+ concentration decreased in both experiments indicating that water absorption occurred in both experiments. Removal of the stopper did not cause any significant changes in the parameters measured. Vasopressin concentration fell immediately on the sight of water, and then continued to decrease. It is concluded that in the sheep, voluntary drinking is followed by an immediate and substantial absorption of water from the rumen.

  16. Effect of bromochloromethane on methane emission, rumen fermentation pattern, milk yield, and fatty acid profile in lactating dairy goats.

    PubMed

    Abecia, L; Toral, P G; Martín-García, A I; Martínez, G; Tomkins, N W; Molina-Alcaide, E; Newbold, C J; Yáñez-Ruiz, D R

    2012-04-01

    Several technologies have been tested to reduce enteric methanogenesis, but very few have been successfully used in practical conditions for livestock. Furthermore, the consequences of reduced rumen methane production on animal performance and milk quality are poorly understood. The aim of this work was to investigate the effect of feeding bromochloromethane (BCM), a halogenated aliphatic hydrocarbon with potential antimethanogenic activity, to dairy goats on rumen methane production, fermentation pattern, the abundance of major microbial groups, and on animal performance and milk composition. Eighteen goats were allocated to 2 experimental groups of 9 animals each: treated (BCM+) or not (BCM-) with 0.30 g of BCM/100 kg of body weight per day. The BCM was administered per os in 2 equal doses per day from parturition to 2 wk postweaning (10 wk). After weaning, methane emissions were recorded over 2 consecutive days (d 57 and 58 on treatment) in polycarbonate chambers. On d 59, individual rumen fluid samples were collected for volatile fatty acid (VFA) analysis and quantification of bacterial, protozoal, and archaeal numbers by real-time PCR. On d 69 and 70, daily milk production was recorded and samples were collected for determination of fat, protein, lactose, casein, and total solids concentration by infrared spectrophotometry, and fatty acid composition by gas chromatography. Treatment with BCM reduced methane production by 33% (21.6 vs. 14.4 L/kg of DMI) compared with nontreated animals, although it did not affect the abundance of rumen bacteria, protozoa, and total methanogenic archaea. The observed improvement in the efficiency of digestive processes was accompanied by a 36% increase in milk yield, probably due to the more propionic type of rumen fermentation and an increase in VFA production. The increase in milk yield was not accompanied by any changes in the concentrations or yields of fat, protein, or lactose. Despite the substantial decrease in methane

  17. Rumen bacterial diversity of 80 to 110-day-old goats using 16S rRNA sequencing.

    PubMed

    Han, Xufeng; Yang, Yuxin; Yan, Hailong; Wang, Xiaolong; Qu, Lei; Chen, Yulin

    2015-01-01

    The ability of rumen microorganisms to use fibrous plant matter plays an important role in ruminant animals; however, little information about rumen colonization by microbial populations after weaning has been reported. In this study, high-throughput sequencing was used to investigate the establishment of this microbial population in 80 to 110-day-old goats. Illumina sequencing of goat rumen samples yielded 101,356,610 nucleotides that were assembled into 256,868 reads with an average read length of 394 nucleotides. Taxonomic analysis of metagenomic reads indicated that the predominant phyla were distinct at different growth stages. The phyla Firmicutes and Synergistetes were predominant in samples taken from 80 to 100-day-old goats, but Bacteroidetes and Firmicutes became the most abundant phyla in samples from 110-day-old animals. There was a remarkable variation in the microbial populations with age; Firmicutes and Synergistetes decreased after weaning, but Bacteroidetes and Proteobacteria increased from 80 to 110 day of age. These findings suggested that colonization of the rumen by microorganisms is related to their function in the rumen digestive system. These results give a better understanding of the role of rumen microbes and the establishment of the microbial population, which help to maintain the host's health and improve animal performance.

  18. Rumen bacterial diversity of 80 to 110-day-old goats using 16S rRNA sequencing.

    PubMed

    Han, Xufeng; Yang, Yuxin; Yan, Hailong; Wang, Xiaolong; Qu, Lei; Chen, Yulin

    2015-01-01

    The ability of rumen microorganisms to use fibrous plant matter plays an important role in ruminant animals; however, little information about rumen colonization by microbial populations after weaning has been reported. In this study, high-throughput sequencing was used to investigate the establishment of this microbial population in 80 to 110-day-old goats. Illumina sequencing of goat rumen samples yielded 101,356,610 nucleotides that were assembled into 256,868 reads with an average read length of 394 nucleotides. Taxonomic analysis of metagenomic reads indicated that the predominant phyla were distinct at different growth stages. The phyla Firmicutes and Synergistetes were predominant in samples taken from 80 to 100-day-old goats, but Bacteroidetes and Firmicutes became the most abundant phyla in samples from 110-day-old animals. There was a remarkable variation in the microbial populations with age; Firmicutes and Synergistetes decreased after weaning, but Bacteroidetes and Proteobacteria increased from 80 to 110 day of age. These findings suggested that colonization of the rumen by microorganisms is related to their function in the rumen digestive system. These results give a better understanding of the role of rumen microbes and the establishment of the microbial population, which help to maintain the host's health and improve animal performance. PMID:25700157

  19. Development of microbial spoilage and lipid and protein oxidation in rabbit meat.

    PubMed

    Nakyinsige, K; Sazili, A Q; Aghwan, Z A; Zulkifli, I; Goh, Y M; Abu Bakar, F; Sarah, S A

    2015-10-01

    This experiment aimed to determine microbial spoilage and lipid and protein oxidation during aerobic refrigerated (4°C) storage of rabbit meat. Forty male New Zealand white rabbits were slaughtered according to the Halal slaughter procedure. The hind limbs were used for microbial analysis while the Longissimus lumborum m. was used for determination of lipid and protein oxidation. Bacterial counts generally increased with aging time and the limit for fresh meat (10(8)cfu/g) was reached at d 7 postmortem. Significant differences in malondialdehyde content were observed after 3d of storage. The thiol concentration significantly decreased with increase in aging time. The band intensities of myosin heavy chain and troponin T significantly reduced with increased refrigerated storage while actin remained relatively stable. This study thus proposes protein oxidation as a potential deteriorative change in refrigerated rabbit meat along with microbial spoilage and lipid oxidation. PMID:26115345

  20. Development of microbial spoilage and lipid and protein oxidation in rabbit meat.

    PubMed

    Nakyinsige, K; Sazili, A Q; Aghwan, Z A; Zulkifli, I; Goh, Y M; Abu Bakar, F; Sarah, S A

    2015-10-01

    This experiment aimed to determine microbial spoilage and lipid and protein oxidation during aerobic refrigerated (4°C) storage of rabbit meat. Forty male New Zealand white rabbits were slaughtered according to the Halal slaughter procedure. The hind limbs were used for microbial analysis while the Longissimus lumborum m. was used for determination of lipid and protein oxidation. Bacterial counts generally increased with aging time and the limit for fresh meat (10(8)cfu/g) was reached at d 7 postmortem. Significant differences in malondialdehyde content were observed after 3d of storage. The thiol concentration significantly decreased with increase in aging time. The band intensities of myosin heavy chain and troponin T significantly reduced with increased refrigerated storage while actin remained relatively stable. This study thus proposes protein oxidation as a potential deteriorative change in refrigerated rabbit meat along with microbial spoilage and lipid oxidation.

  1. A Metagenomics Approach to Evaluate the Impact of Dietary Supplementation with Ascophyllum nodosum or Laminaria digitata on Rumen Function in Rusitec Fermenters

    PubMed Central

    Belanche, Alejandro; Jones, Eleanor; Parveen, Ifat; Newbold, Charles J.

    2016-01-01

    There is an increasing need to identify alternative feeds for livestock that do not compete with foods for humans. Seaweed might provide such a resource, but there is limited information available on its value as an animal feed. Here we use a multi-omics approach to investigate the value of two brown seaweeds, Ascophyllum nodosum (ASC) and Laminaria digitata (LAM), as alternative feeds for ruminants. These seaweeds were supplemented at 5% inclusion rate into a control diet (CON) in a rumen simulation fermenter. The seaweeds had no substantial effect on rumen fermentation, feed degradability or methane emissions. Concentrations of total bacteria, anaerobic fungi, biodiversity indices and abundances of the main bacterial and methanogen genera were also unaffected. However, species-specific effects of brown seaweed on the rumen function were noted: ASC promoted a substantial decrease in N degradability (−24%) due to its high phlorotannins content. Canonical correspondence analysis of the bacterial community revealed that low N availability led to a change in the structure of the bacterial community. ASC also decreased the concentration of Escherichia coli O157:H7 post-inoculation. In contrast, LAM which has a much lower phlorotannin content did not cause detrimental effects on N degradability nor modified the structure of the bacterial community in comparison to CON. This adaptation of the microbial community to LAM diets led to a greater microbial ability to digest xylan (+70%) and carboxy-methyl-cellulose (+41%). These differences among brown seaweeds resulted in greater microbial protein synthesis (+15%) and non-ammonia N flow (+11%) in LAM than in ASC diets and thus should led to a greater amino acid supply to the intestine of the animal. In conclusion, it was demonstrated that incorporation of brown seaweed into the diet can be considered as a suitable nutritional strategy for ruminants; however, special care must be taken with those seaweeds with high

  2. A Metagenomics Approach to Evaluate the Impact of Dietary Supplementation with Ascophyllum nodosum or Laminaria digitata on Rumen Function in Rusitec Fermenters.

    PubMed

    Belanche, Alejandro; Jones, Eleanor; Parveen, Ifat; Newbold, Charles J

    2016-01-01

    There is an increasing need to identify alternative feeds for livestock that do not compete with foods for humans. Seaweed might provide such a resource, but there is limited information available on its value as an animal feed. Here we use a multi-omics approach to investigate the value of two brown seaweeds, Ascophyllum nodosum (ASC) and Laminaria digitata (LAM), as alternative feeds for ruminants. These seaweeds were supplemented at 5% inclusion rate into a control diet (CON) in a rumen simulation fermenter. The seaweeds had no substantial effect on rumen fermentation, feed degradability or methane emissions. Concentrations of total bacteria, anaerobic fungi, biodiversity indices and abundances of the main bacterial and methanogen genera were also unaffected. However, species-specific effects of brown seaweed on the rumen function were noted: ASC promoted a substantial decrease in N degradability (-24%) due to its high phlorotannins content. Canonical correspondence analysis of the bacterial community revealed that low N availability led to a change in the structure of the bacterial community. ASC also decreased the concentration of Escherichia coli O157:H7 post-inoculation. In contrast, LAM which has a much lower phlorotannin content did not cause detrimental effects on N degradability nor modified the structure of the bacterial community in comparison to CON. This adaptation of the microbial community to LAM diets led to a greater microbial ability to digest xylan (+70%) and carboxy-methyl-cellulose (+41%). These differences among brown seaweeds resulted in greater microbial protein synthesis (+15%) and non-ammonia N flow (+11%) in LAM than in ASC diets and thus should led to a greater amino acid supply to the intestine of the animal. In conclusion, it was demonstrated that incorporation of brown seaweed into the diet can be considered as a suitable nutritional strategy for ruminants; however, special care must be taken with those seaweeds with high

  3. Microbial protein: future sustainable food supply route with low environmental footprint.

    PubMed

    Matassa, Silvio; Boon, Nico; Pikaar, Ilje; Verstraete, Willy

    2016-09-01

    Microbial biotechnology has a long history of producing feeds and foods. The key feature of today's market economy is that protein production by conventional agriculture based food supply chains is becoming a major issue in terms of global environmental pollution such as diffuse nutrient and greenhouse gas emissions, land use and water footprint. Time has come to re-assess the current potentials of producing protein-rich feed or food additives in the form of algae, yeasts, fungi and plain bacterial cellular biomass, producible with a lower environmental footprint compared with other plant or animal-based alternatives. A major driver is the need to no longer disintegrate but rather upgrade a variety of low-value organic and inorganic side streams in our current non-cyclic economy. In this context, microbial bioconversions of such valuable matters to nutritive microbial cells and cell components are a powerful asset. The worldwide market of animal protein is of the order of several hundred million tons per year, that of plant protein several billion tons of protein per year; hence, the expansion of the production of microbial protein does not pose disruptive challenges towards the process of the latter. Besides protein as nutritive compounds, also other cellular components such as lipids (single cell oil), polyhydroxybuthyrate, exopolymeric saccharides, carotenoids, ectorines, (pro)vitamins and essential amino acids can be of value for the growing domain of novel nutrition. In order for microbial protein as feed or food to become a major and sustainable alternative, addressing the challenges of creating awareness and achieving public and broader regulatory acceptance are real and need to be addressed with care and expedience. PMID:27389856

  4. Microbial Protein-Protein Interactions (MiPPI) Data from the Genomics: GTL Center for Molecular and Cellular Systems (CMCS)

    DOE Data Explorer

    The Genomic Science Center for Molecular and Cellular Systems (CMCS), established in 2002, seeks to identify and characterize the complete set of protein complexes within a cell to provide a mechanistic basis for the understanding of biochemical functions. The CMCS is anchored at ORNL and PNNL. CMCS initially focused on the identification and characterization of protein complexes in two microbial systems,Rhodopseudomonas palustris (R. palustris) and Shewanella oneidensis (S. oneidensis). These two organisms have also been the focus of major DOE Genomic Science/Microbial Cell Program (MCP) projects. To develop an approach for identifying the diverse types of complexes present in microbial organisms, CMCS incorporates a number of molecular biology, microbiology, analytical and computational tools in an integrated pipeline.

  5. Interaction of unsaturated fat or coconut oil with monensin in lactating dairy cows fed 12 times daily. I. Protozoal abundance, nutrient digestibility, and microbial protein flow to the omasum.

    PubMed

    Reveneau, C; Karnati, S K R; Oelker, E R; Firkins, J L

    2012-04-01

    Monensin (tradename: Rumensin) should reduce the extent of amino acid deamination in the rumen, and supplemental fat should decrease protozoal abundance and intraruminal N recycling. Because animal-vegetable (AV) fat can be biohydrogenated in the rumen and decrease its effectiveness as an anti-protozoal agent, we included diets supplemented with coconut oil (CNO) to inhibit protozoa. In a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of treatments, 6 rumen-cannulated cows were fed diets without or with Rumensin (12 g/909 kg) and either no fat (control), 5% AV fat, or 5% CNO. The log10 concentrations (cells/mL) of total protozoa were not different between control (5.97) and AV fat (5.95) but were decreased by CNO (4.79; main effect of fat source). Entodinium and Dasytricha decreased as a proportion of total cells from feeding CNO, whereas Epidinium was unchanged in total abundance and thus increased proportionately. Total volatile fatty acid concentration was not affected by diet, but the acetate:propionate ratio decreased for CNO (1.85) versus control (2.95) or AV fat (2.58). Feeding CNO (23.8%) decreased ruminal neutral detergent fiber digestibility compared with control (31.1%) and AV fat (30.5%). The total-tract digestibility of NDF was lower for CNO (45.8%) versus control (57.0%) and AV fat (54.6%), with no difference in apparent organic matter digestibility (averaging 69.8%). The omasal flows of microbial N and non-ammonia N were lower for CNO versus control and AV fat, but efficiency of microbial protein synthesis was not affected. The dry matter intake was 4.5 kg/d lower with CNO, which decreased milk production by 3.1 kg/d. Main effect means of dry matter intake and milk yield tended to decrease by 0.7 and 1.2 kg/d, respectively, when Rumensin was added. Both percentage and production of milk fat decreased for CNO (main effect of fat source). An interaction was observed such that AV decreased milk fat yield more when combined with Rumensin

  6. Determination of Microbial Growth by Protein Assay in an Air-Cathode Single Chamber Microbial Fuel Cell.

    PubMed

    Li, Na; Kakarla, Ramesh; Moon, Jung Mi; Min, Booki

    2015-07-01

    Microbial fuel cells (MFCs) have gathered attention as a novel bioenergy technology to simultaneously treat wastewater with less sludge production than the conventional activated sludge system. In two different operations of the MFC and aerobic process, microbial growth was determined by the protein assay method and their biomass yields using real wastewater were compared. The biomass yield on the anode electrode of the MFC was 0.02 g-COD-cell/g- COD-substrate and the anolyte planktonic biomass was 0.14 g-COD-cell/g-COD-substrate. An MFC without anode electrode resulted in the biomass yield of 0.07 ± 0.03 g-COD-cell/g-COD-substrate, suggesting that oxygen diffusion from the cathode possibly supported the microbial growth. In a comparative test, the biomass yield under aerobic environment was 0.46 ± 0.07 g-COD-cell/g-COD-substrate, which was about 3 times higher than the total biomass value in the MFC operation.

  7. Comparative evaluation of rumen metagenome community using qPCR and MG-RAST.

    PubMed

    Nathani, Neelam M; Patel, Amrutlal K; Dhamannapatil, Prakash S; Kothari, Ramesh K; Singh, Krishna M; Joshi, Chaitanya G

    2013-01-01

    Microbial profiling of metagenome communities have been studied extensively using MG-RAST and other related metagenome annotation databases. Although, database based taxonomic profiling provides snapshots of the metagenome architecture, their reliability needs to be validated through more accurate methods. Here, we performed qPCR based absolute quantitation of selected rumen microbes in the liquid and solid fraction of the rumen fluid of river buffalo adapted to varying proportion of concentrate to green or dry roughages and compared with the MG-RAST based annotation of the metagenomes sequences of 16S r-DNA amplicons and high throughput shotgun sequencing. Animals were adapted to roughage-to-concentrate ratio in the proportion of 50:50, 75:25 and 100:00, respectively for six weeks. At the end of each treatment, rumen fluid was collected at 3 h post feeding. qPCR revealed that the relative abundance of Prevotella bryantii was higher, followed by the two cellulolytic bacteria Fibrobacter succinogens and Ruminococcus flavefaciens that accounted up to 1.33% and 0.78% of the total rumen bacteria, respectively. While, Selenomonas ruminantium and archaea Methanomicrobiales were lower in microbial population in the rumen of buffalo. There was no statistically significant difference between the enumerations shown by qPCR and analysis of the shotgun sequencing data by MG-RAST except for Prevotella. These results indicate the variations in abundance of different microbial species in buffalo rumen under varied feeding regimes as well as in different fractions of rumen liquor, i.e. solid and the liquid. The results also present the reliability of shotgun sequencing to describe metagenome and analysis/annotation by MG-RAST. PMID:24025701

  8. Rumen Bacterial Community Composition in Holstein and Jersey Cows Is Different under Same Dietary Condition and Is Not Affected by Sampling Method

    PubMed Central

    Paz, Henry A.; Anderson, Christopher L.; Muller, Makala J.; Kononoff, Paul J.; Fernando, Samodha C.

    2016-01-01

    The rumen microbial community in dairy cows plays a critical role in efficient milk production. However, there is a lack of data comparing the composition of the rumen bacterial community of the main dairy breeds. This study utilizes 16S rRNA gene sequencing to describe the rumen bacterial community composition in Holstein and Jersey cows fed the same diet by sampling the rumen microbiota via the rumen cannula (Holstein cows) or esophageal tubing (both Holstein and Jersey cows). After collection of the rumen sample via esophageal tubing, particles attached to the strainer were added to the sample to ensure representative sampling of both the liquid and solid fraction of the rumen contents. Alpha diversity metrics, Chao1 and observed OTUs estimates, displayed higher (P = 0.02) bacterial richness in Holstein compared to Jersey cows and no difference (P > 0.70) in bacterial community richness due to sampling method. The principal coordinate analysis displayed distinct clustering of bacterial communities by breed suggesting that Holstein and Jersey cows harbor different rumen bacterial communities. Family level classification of most abundant (>1%) differential OTUs displayed that OTUs from the bacterial families Lachnospiraceae and p-2534-18B5 to be predominant in Holstein cows compared to Jersey cows. Additionally, OTUs belonging to family Prevotellaceae were differentially abundant in the two breeds. Overall, the results from this study suggest that the bacterial community between Holstein and Jersey cows differ and that esophageal tubing with collection of feed particles associated with the strainer provides a representative rumen sample similar to a sample collected via the rumen cannula. Thus, in future studies esophageal tubing with addition of retained particles can be used to collect rumen samples reducing the cost of cannulation and increasing the number of animals used in microbiome investigations, thus increasing the statistical power of rumen microbial

  9. Rumen Bacterial Community Composition in Holstein and Jersey Cows Is Different under Same Dietary Condition and Is Not Affected by Sampling Method.

    PubMed

    Paz, Henry A; Anderson, Christopher L; Muller, Makala J; Kononoff, Paul J; Fernando, Samodha C

    2016-01-01

    The rumen microbial community in dairy cows plays a critical role in efficient milk production. However, there is a lack of data comparing the composition of the rumen bacterial community of the main dairy breeds. This study utilizes 16S rRNA gene sequencing to describe the rumen bacterial community composition in Holstein and Jersey cows fed the same diet by sampling the rumen microbiota via the rumen cannula (Holstein cows) or esophageal tubing (both Holstein and Jersey cows). After collection of the rumen sample via esophageal tubing, particles attached to the strainer were added to the sample to ensure representative sampling of both the liquid and solid fraction of the rumen contents. Alpha diversity metrics, Chao1 and observed OTUs estimates, displayed higher (P = 0.02) bacterial richness in Holstein compared to Jersey cows and no difference (P > 0.70) in bacterial community richness due to sampling method. The principal coordinate analysis displayed distinct clustering of bacterial communities by breed suggesting that Holstein and Jersey cows harbor different rumen bacterial communities. Family level classification of most abundant (>1%) differential OTUs displayed that OTUs from the bacterial families Lachnospiraceae and p-2534-18B5 to be predominant in Holstein cows compared to Jersey cows. Additionally, OTUs belonging to family Prevotellaceae were differentially abundant in the two breeds. Overall, the results from this study suggest that the bacterial community between Holstein and Jersey cows differ and that esophageal tubing with collection of feed particles associated with the strainer provides a representative rumen sample similar to a sample collected via the rumen cannula. Thus, in future studies esophageal tubing with addition of retained particles can be used to collect rumen samples reducing the cost of cannulation and increasing the number of animals used in microbiome investigations, thus increasing the statistical power of rumen microbial

  10. Rumen Bacterial Community Composition in Holstein and Jersey Cows Is Different under Same Dietary Condition and Is Not Affected by Sampling Method.

    PubMed

    Paz, Henry A; Anderson, Christopher L; Muller, Makala J; Kononoff, Paul J; Fernando, Samodha C

    2016-01-01

    The rumen microbial community in dairy cows plays a critical role in efficient milk production. However, there is a lack of data comparing the composition of the rumen bacterial community of the main dairy breeds. This study utilizes 16S rRNA gene sequencing to describe the rumen bacterial community composition in Holstein and Jersey cows fed the same diet by sampling the rumen microbiota via the rumen cannula (Holstein cows) or esophageal tubing (both Holstein and Jersey cows). After collection of the rumen sample via esophageal tubing, particles attached to the strainer were added to the sample to ensure representative sampling of both the liquid and solid fraction of the rumen contents. Alpha diversity metrics, Chao1 and observed OTUs estimates, displayed higher (P = 0.02) bacterial richness in Holstein compared to Jersey cows and no difference (P > 0.70) in bacterial community richness due to sampling method. The principal coordinate analysis displayed distinct clustering of bacterial communities by breed suggesting that Holstein and Jersey cows harbor different rumen bacterial communities. Family level classification of most abundant (>1%) differential OTUs displayed that OTUs from the bacterial families Lachnospiraceae and p-2534-18B5 to be predominant in Holstein cows compared to Jersey cows. Additionally, OTUs belonging to family Prevotellaceae were differentially abundant in the two breeds. Overall, the results from this study suggest that the bacterial community between Holstein and Jersey cows differ and that esophageal tubing with collection of feed particles associated with the strainer provides a representative rumen sample similar to a sample collected via the rumen cannula. Thus, in future studies esophageal tubing with addition of retained particles can be used to collect rumen samples reducing the cost of cannulation and increasing the number of animals used in microbiome investigations, thus increasing the statistical power of rumen microbial

  11. Effects of Different Protein Supplements on Omasal Nutrient Flow and Microbial Protein Synthesis in Lactating Dairy Cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eight ruminally cannulated Holstein cows that were part of a larger lactation trial were used in 2 replicated 4 x 4 Latin squares to quantify effects of supplementing protein as urea, solvent soybean meal (SSBM), cottonseed meal (CSM), or canola meal (CM) on omasal nutrient flows and microbial prote...

  12. Interaction of molasses and monensin in alfalfa hay- or corn silage-based diets on rumen fermentation, total tract digestibility, and milk production by Holstein cows.

    PubMed

    Oelker, E R; Reveneau, C; Firkins, J L

    2009-01-01

    Sugar supplementation can stimulate rumen microbial growth and possibly fiber digestibility; however, excess ruminal carbohydrate availability relative to rumen-degradable protein (RDP) can promote energy spilling by microbes, decrease rumen pH, or depress fiber digestibility. Both RDP supply and rumen pH might be altered by forage source and monensin. Therefore, the objective of this study was to evaluate interactions of a sugar source (molasses) with monensin and 2 forage sources on rumen fermentation, total tract digestibility, and production and fatty acid composition of milk. Seven ruminally cannulated lactating Holstein cows were used in a 5 x 7 incomplete Latin square design with five 28-d periods. Four corn silage diets consisted of 1) control (C), 2) 2.6% molasses (M), 3) 2.6% molasses plus 0.45% urea (MU), or 4) 2.6% molasses plus 0.45% urea plus monensin sodium (Rumensin, at the intermediate dosage from the label, 16 g/909 kg of dry matter; MUR). Three chopped alfalfa hay diets consisted of 1) control (C), 2) 2.6% molasses (M), or 3) 2.6% molasses plus Rumensin (MR). Urea was added to corn silage diets to provide RDP comparable to alfalfa hay diets with no urea. Corn silage C and M diets were balanced to have 16.2% crude protein; and the remaining diets, 17.2% crude protein. Dry matter intake was not affected by treatment, but there was a trend for lower milk production in alfalfa hay diets compared with corn silage diets. Despite increased total volatile fatty acid and acetate concentrations in the rumen, total tract organic matter digestibility was lower for alfalfa hay-fed cows. Rumensin did not affect volatile fatty acid concentrations but decreased milk fat from 3.22 to 2.72% in corn silage diets but less in alfalfa hay diets. Medium-chain milk fatty acids (% of total fat) were lower for alfalfa hay compared with corn silage diets, and short-chain milk fatty acids tended to decrease when Rumensin was added. In whole rumen contents, concentrations of

  13. Functional metagenomics unveils a multifunctional glycosyl hydrolase from the family 43 catalysing the breakdown of plant polymers in the calf rumen.

    PubMed

    Ferrer, Manuel; Ghazi, Azam; Beloqui, Ana; Vieites, José María; López-Cortés, Nieves; Marín-Navarro, Julia; Nechitaylo, Taras Y; Guazzaroni, María-Eugenia; Polaina, Julio; Waliczek, Agnes; Chernikova, Tatyana N; Reva, Oleg N; Golyshina, Olga V; Golyshin, Peter N

    2012-01-01

    Microbial communities from cow rumen are known for their ability to degrade diverse plant polymers at high rates. In this work, we identified 15 hydrolases through an activity-centred metagenome analysis of a fibre-adherent microbial community from dairy cow rumen. Among them, 7 glycosyl hydrolases (GHs) and 1 feruloyl esterase were successfully cloned, expressed, purified and characterised. The most striking result was a protein of GH family 43 (GHF43), hereinafter designated as R_09-02, which had characteristics very distinct from the other proteins in this family with mono-functional β-xylosidase, α-xylanase, α-L-arabinase and α-L-arabinofuranosidase activities. R_09-02 is the first multifunctional enzyme to exhibit β-1,4 xylosidase, α-1,5 arabinofur(pyr)anosidase, β-1,4 lactase, α-1,6 raffinase, α-1,6 stachyase, β-galactosidase and α-1,4 glucosidase activities. The R_09-02 protein appears to originate from the chromosome of a member of Clostridia, a class of phylum Firmicutes, members of which are highly abundant in ruminal environment. The evolution of R_09-02 is suggested to be driven from the xylose- and arabinose-specific activities, typical for GHF43 members, toward a broader specificity to the glucose- and galactose-containing components of lignocellulose. The apparent capability of enzymes from the GHF43 family to utilise xylose-, arabinose-, glucose- and galactose-containing oligosaccharides has thus far been neglected by, or could not be predicted from, genome and metagenome sequencing data analyses. Taking into account the abundance of GHF43-encoding gene sequences in the rumen (up to 7% of all GH-genes) and the multifunctional phenotype herein described, our findings suggest that the ecological role of this GH family in the digestion of ligno-cellulosic matter should be significantly reconsidered. PMID:22761666

  14. Metagenomic Insights into the Carbohydrate-Active Enzymes Carried by the Microorganisms Adhering to Solid Digesta in the Rumen of Cows

    PubMed Central

    Wang, Lingling; Hatem, Ayat; Catalyurek, Umit V.; Morrison, Mark; Yu, Zhongtang

    2013-01-01

    The ruminal microbial community is a unique source of enzymes that underpin the conversion of cellulosic biomass. In this study, the microbial consortia adherent on solid digesta in the rumen of Jersey cattle were subjected to an activity-based metagenomic study to explore the genetic diversity of carbohydrolytic enzymes in Jersey cows, with a particular focus on cellulases and xylanases. Pyrosequencing and bioinformatic analyses of 120 carbohydrate-active fosmids identified genes encoding 575 putative Carbohydrate-Active Enzymes (CAZymes) and proteins putatively related to transcriptional regulation, transporters, and signal transduction coupled with polysaccharide degradation and metabolism. Most of these genes shared little similarity to sequences archived in databases. Genes that were predicted to encode glycoside hydrolases (GH) involved in xylan and cellulose hydrolysis (e.g., GH3, 5, 9, 10, 39 and 43) were well represented. A new subfamily (S-8) of GH5 was identified from contigs assigned to Firmicutes. These subfamilies of GH5 proteins also showed significant phylum-dependent distribution. A number of polysaccharide utilization loci (PULs) were found, and two of them contained genes encoding Sus-like proteins and cellulases that have not been reported in previous metagenomic studies of samples from the rumens of cows or other herbivores. Comparison with the large metagenomic datasets previously reported of other ruminant species (or cattle breeds) and wallabies showed that the rumen microbiome of Jersey cows might contain differing CAZymes. Future studies are needed to further explore how host genetics and diets affect the diversity and distribution of CAZymes and utilization of plant cell wall materials. PMID:24223817

  15. Functional Metagenomics Unveils a Multifunctional Glycosyl Hydrolase from the Family 43 Catalysing the Breakdown of Plant Polymers in the Calf Rumen

    PubMed Central

    Vieites, José María; López-Cortés, Nieves; Marín-Navarro, Julia; Nechitaylo, Taras Y.; Guazzaroni, María-Eugenia; Polaina, Julio; Waliczek, Agnes; Chernikova, Tatyana N.; Reva, Oleg N.; Golyshina, Olga V.; Golyshin, Peter N.

    2012-01-01

    Microbial communities from cow rumen are known for their ability to degrade diverse plant polymers at high rates. In this work, we identified 15 hydrolases through an activity-centred metagenome analysis of a fibre-adherent microbial community from dairy cow rumen. Among them, 7 glycosyl hydrolases (GHs) and 1 feruloyl esterase were successfully cloned, expressed, purified and characterised. The most striking result was a protein of GH family 43 (GHF43), hereinafter designated as R_09-02, which had characteristics very distinct from the other proteins in this family with mono-functional β-xylosidase, α-xylanase, α-L-arabinase and α-L-arabinofuranosidase activities. R_09-02 is the first multifunctional enzyme to exhibit β-1,4 xylosidase, α-1,5 arabinofur(pyr)anosidase, β-1,4 lactase, α-1,6 raffinase, α-1,6 stachyase, β-galactosidase and α-1,4 glucosidase activities. The R_09-02 protein appears to originate from the chromosome of a member of Clostridia, a class of phylum Firmicutes, members of which are highly abundant in ruminal environment. The evolution of R_09-02 is suggested to be driven from the xylose- and arabinose-specific activities, typical for GHF43 members, toward a broader specificity to the glucose- and galactose-containing components of lignocellulose. The apparent capability of enzymes from the GHF43 family to utilise xylose-, arabinose-, glucose- and galactose-containing oligosaccharides has thus far been neglected by, or could not be predicted from, genome and metagenome sequencing data analyses. Taking into account the abundance of GHF43-encoding gene sequences in the rumen (up to 7% of all GH-genes) and the multifunctional phenotype herein described, our findings suggest that the ecological role of this GH family in the digestion of ligno-cellulosic matter should be significantly reconsidered. PMID:22761666

  16. Functional metagenomics unveils a multifunctional glycosyl hydrolase from the family 43 catalysing the breakdown of plant polymers in the calf rumen.

    PubMed

    Ferrer, Manuel; Ghazi, Azam; Beloqui, Ana; Vieites, José María; López-Cortés, Nieves; Marín-Navarro, Julia; Nechitaylo, Taras Y; Guazzaroni, María-Eugenia; Polaina, Julio; Waliczek, Agnes; Chernikova, Tatyana N; Reva, Oleg N; Golyshina, Olga V; Golyshin, Peter N

    2012-01-01

    Microbial communities from cow rumen are known for their ability to degrade diverse plant polymers at high rates. In this work, we identified 15 hydrolases through an activity-centred metagenome analysis of a fibre-adherent microbial community from dairy cow rumen. Among them, 7 glycosyl hydrolases (GHs) and 1 feruloyl esterase were successfully cloned, expressed, purified and characterised. The most striking result was a protein of GH family 43 (GHF43), hereinafter designated as R_09-02, which had characteristics very distinct from the other proteins in this family with mono-functional β-xylosidase, α-xylanase, α-L-arabinase and α-L-arabinofuranosidase activities. R_09-02 is the first multifunctional enzyme to exhibit β-1,4 xylosidase, α-1,5 arabinofur(pyr)anosidase, β-1,4 lactase, α-1,6 raffinase, α-1,6 stachyase, β-galactosidase and α-1,4 glucosidase activities. The R_09-02 protein appears to originate from the chromosome of a member of Clostridia, a class of phylum Firmicutes, members of which are highly abundant in ruminal environment. The evolution of R_09-02 is suggested to be driven from the xylose- and arabinose-specific activities, typical for GHF43 members, toward a broader specificity to the glucose- and galactose-containing components of lignocellulose. The apparent capability of enzymes from the GHF43 family to utilise xylose-, arabinose-, glucose- and galactose-containing oligosaccharides has thus far been neglected by, or could not be predicted from, genome and metagenome sequencing data analyses. Taking into account the abundance of GHF43-encoding gene sequences in the rumen (up to 7% of all GH-genes) and the multifunctional phenotype herein described, our findings suggest that the ecological role of this GH family in the digestion of ligno-cellulosic matter should be significantly reconsidered.

  17. Metagenomic insights into the carbohydrate-active enzymes carried by the microorganisms adhering to solid digesta in the rumen of cows.

    PubMed

    Wang, Lingling; Hatem, Ayat; Catalyurek, Umit V; Morrison, Mark; Yu, Zhongtang

    2013-01-01

    The ruminal microbial community is a unique source of enzymes that underpin the conversion of cellulosic biomass. In this study, the microbial consortia adherent on solid digesta in the rumen of Jersey cattle were subjected to an activity-based metagenomic study to explore the genetic diversity of carbohydrolytic enzymes in Jersey cows, with a particular focus on cellulases and xylanases. Pyrosequencing and bioinformatic analyses of 120 carbohydrate-active fosmids identified genes encoding 575 putative Carbohydrate-Active Enzymes (CAZymes) and proteins putatively related to transcriptional regulation, transporters, and signal transduction coupled with polysaccharide degradation and metabolism. Most of these genes shared little similarity to sequences archived in databases. Genes that were predicted to encode glycoside hydrolases (GH) involved in xylan and cellulose hydrolysis (e.g., GH3, 5, 9, 10, 39 and 43) were well represented. A new subfamily (S-8) of GH5 was identified from contigs assigned to Firmicutes. These subfamilies of GH5 proteins also showed significant phylum-dependent distribution. A number of polysaccharide utilization loci (PULs) were found, and two of them contained genes encoding Sus-like proteins and cellulases that have not been reported in previous metagenomic studies of samples from the rumens of cows or other herbivores. Comparison with the large metagenomic datasets previously reported of other ruminant species (or cattle breeds) and wallabies showed that the rumen microbiome of Jersey cows might contain differing CAZymes. Future studies are needed to further explore how host genetics and diets affect the diversity and distribution of CAZymes and utilization of plant cell wall materials. PMID:24223817

  18. Metagenomic insights into the carbohydrate-active enzymes carried by the microorganisms adhering to solid digesta in the rumen of cows.

    PubMed

    Wang, Lingling; Hatem, Ayat; Catalyurek, Umit V; Morrison, Mark; Yu, Zhongtang

    2013-01-01

    The ruminal microbial community is a unique source of enzymes that underpin the conversion of cellulosic biomass. In this study, the microbial consortia adherent on solid digesta in the rumen of Jersey cattle were subjected to an activity-based metagenomic study to explore the genetic diversity of carbohydrolytic enzymes in Jersey cows, with a particular focus on cellulases and xylanases. Pyrosequencing and bioinformatic analyses of 120 carbohydrate-active fosmids identified genes encoding 575 putative Carbohydrate-Active Enzymes (CAZymes) and proteins putatively related to transcriptional regulation, transporters, and signal transduction coupled with polysaccharide degradation and metabolism. Most of these genes shared little similarity to sequences archived in databases. Genes that were predicted to encode glycoside hydrolases (GH) involved in xylan and cellulose hydrolysis (e.g., GH3, 5, 9, 10, 39 and 43) were well represented. A new subfamily (S-8) of GH5 was identified from contigs assigned to Firmicutes. These subfamilies of GH5 proteins also showed significant phylum-dependent distribution. A number of polysaccharide utilization loci (PULs) were found, and two of them contained genes encoding Sus-like proteins and cellulases that have not been reported in previous metagenomic studies of samples from the rumens of cows or other herbivores. Comparison with the large metagenomic datasets previously reported of other ruminant species (or cattle breeds) and wallabies showed that the rumen microbiome of Jersey cows might contain differing CAZymes. Future studies are needed to further explore how host genetics and diets affect the diversity and distribution of CAZymes and utilization of plant cell wall materials.

  19. Structure and Function of Microbial Metal-Reduction Proteins

    SciTech Connect

    Xu, Ying; Crawford, Oakly H.; Xu, Dong; Larimer, Frank W.; Uberbacher, Edward C.; Zhou, Jizhong

    2009-09-02

    In this project, we proposed (i) identification of metal-reduction genes, (ii) development of new threading techniques and (iii) fold recognition and structure prediction of metal-reduction proteins. However, due to the reduction of the budget, we revised our plan to focus on two specific aims of (i) developing a new threading-based protein structure prediction method, and (ii) developing an expert system for protein structure prediction.

  20. Supplementation with non-fibrous carbohydrates reduced fiber digestibility and did not improve microbial protein synthesis in sheep fed fresh forage of two nutritive values.

    PubMed

    Tebot, I; Cajarville, C; Repetto, J L; Cirio, A

    2012-04-01

    To determine whether non-fibrous carbohydrate (NFC) supplementation improves fiber digestibility and microbial protein synthesis, 18 Corriedale ewes with a fixed intake level (40 g dry matter (DM)/kg BW0.75) were assigned to three (n = 6) diets: F = 100% fresh temperate forage, FG = 70% forage + 30% barley grain and FGM = 70% forage + 15% barley grain + 15% molasses-based product (MBP, Kalori 3000). Two experimental periods were carried out, with late (P1) and early (P2) vegetative stage forage. For P2, ewes were fitted with ruminal catheters. Forage was distributed at 0900 h, 1300 h, 1800 h and 2300 h, and supplement added at 0900 h and 1800 h meals. Digestibility of the different components of the diets, retained N and rumen microbial protein synthesis were determined. At the end of P2, ruminal pH and N-NH3 concentration were determined hourly for 24 h. Supplementation increased digestibility of DM (P < 0.001) and organic matter (OM; P < 0.001) and reduced NDF digestibility (P = 0.043) in both periods, with greater values in P2 (P = 0.008) for the three diets. Daily mean ruminal pH differed (P < 0.05) among treatments: 6.33 (F), 6.15 (FG) and 6.51 (FGM). The high pH in FGM was attributed to Ca(OH)2 in MBP. Therefore, the decreased fiber digestibility in supplemented diets could not be attributed to pH changes. The mean ruminal concentration of N-NH3 was 18.0 mg/dl, without differences among treatments or sampling hours. Microbial protein synthesis was greater in P2 (8.0 g/day) than in P1 (6.1 g/day; P = 0.006), but treatments did not enhance this parameter. The efficiency of protein synthesis tended to be lower in supplemented groups (16.4, 13.9 and 13.4 in P1, and 20.8, 16.7 and 16.2 g N/kg digestible OM ingested in P2, for F, FG and FGM, respectively; P = 0.07) without differences between supplements. The same tendency was observed for retained N: 2.55, 1.38 and 1.98 in P1, and 2.28, 1.23 and 1.10 g/day in P2, for F, FG and FGM, respectively; P = 0.05). The

  1. Metagenomic Analysis of the Rumen Microbiome of Steers with Wheat-Induced Frothy Bloat

    PubMed Central

    Pitta, D. W.; Pinchak, W. E.; Indugu, N.; Vecchiarelli, B.; Sinha, R.; Fulford, J. D.

    2016-01-01

    Frothy bloat is a serious metabolic disorder that affects stocker cattle grazing hard red winter wheat forage in the Southern Great Plains causing reduced performance, morbidity, and mortality. We hypothesize that a microbial dysbiosis develops in the rumen microbiome of stocker cattle when grazing on high quality winter wheat pasture that predisposes them to frothy bloat risk. In this study, rumen contents were harvested from six cannulated steers grazing hard red winter wheat (three with bloat score “2” and three with bloat score “0”), extracted for genomic DNA and subjected to 16S rDNA and shotgun sequencing on 454/Roche platform. Approximately 1.5 million reads were sequenced, assembled and assigned for phylogenetic and functional annotations. Bacteria predominated up to 84% of the sequences while archaea contributed to nearly 5% of the sequences. The abundance of archaea was higher in bloated animals (P < 0.05) and dominated by Methanobrevibacter. Predominant bacterial phyla were Firmicutes (65%), Actinobacteria (13%), Bacteroidetes (10%), and Proteobacteria (6%) across all samples. Genera from Firmicutes such as Clostridium, Eubacterium, and Butyrivibrio increased (P < 0.05) while Prevotella from Bacteroidetes decreased in bloated samples. Co-occurrence analysis revealed syntrophic associations between bacteria and archaea in non-bloated samples, however; such interactions faded in bloated samples. Functional annotations of assembled reads to Subsystems database revealed the abundance of several metabolic pathways, with carbohydrate and protein metabolism well represented. Assignment of contigs to CaZy database revealed a greater diversity of Glycosyl Hydrolases dominated by oligosaccharide breaking enzymes (>70%) in non-bloated samples. However, the abundance and diversity of CaZymes were greatly reduced in bloated samples indicating the disruption of carbohydrate metabolism. We conclude that mild to moderate frothy bloat results from tradeoffs both

  2. Metagenomic Analysis of the Rumen Microbiome of Steers with Wheat-Induced Frothy Bloat.

    PubMed

    Pitta, D W; Pinchak, W E; Indugu, N; Vecchiarelli, B; Sinha, R; Fulford, J D

    2016-01-01

    Frothy bloat is a serious metabolic disorder that affects stocker cattle grazing hard red winter wheat forage in the Southern Great Plains causing reduced performance, morbidity, and mortality. We hypothesize that a microbial dysbiosis develops in the rumen microbiome of stocker cattle when grazing on high quality winter wheat pasture that predisposes them to frothy bloat risk. In this study, rumen contents were harvested from six cannulated steers grazing hard red winter wheat (three with bloat score "2" and three with bloat score "0"), extracted for genomic DNA and subjected to 16S rDNA and shotgun sequencing on 454/Roche platform. Approximately 1.5 million reads were sequenced, assembled and assigned for phylogenetic and functional annotations. Bacteria predominated up to 84% of the sequences while archaea contributed to nearly 5% of the sequences. The abundance of archaea was higher in bloated animals (P < 0.05) and dominated by Methanobrevibacter. Predominant bacterial phyla were Firmicutes (65%), Actinobacteria (13%), Bacteroidetes (10%), and Proteobacteria (6%) across all samples. Genera from Firmicutes such as Clostridium, Eubacterium, and Butyrivibrio increased (P < 0.05) while Prevotella from Bacteroidetes decreased in bloated samples. Co-occurrence analysis revealed syntrophic associations between bacteria and archaea in non-bloated samples, however; such interactions faded in bloated samples. Functional annotations of assembled reads to Subsystems database revealed the abundance of several metabolic pathways, with carbohydrate and protein metabolism well represented. Assignment of contigs to CaZy database revealed a greater diversity of Glycosyl Hydrolases dominated by oligosaccharide breaking enzymes (>70%) in non-bloated samples. However, the abundance and diversity of CaZymes were greatly reduced in bloated samples indicating the disruption of carbohydrate metabolism. We conclude that mild to moderate frothy bloat results from tradeoffs both within

  3. Determining the amount of rumen-protected methionine supplement that corresponds to the optimal levels of methionine in metabolizable protein for maximizing milk protein production and profit on dairy farms.

    PubMed

    Cho, J; Overton, T R; Schwab, C G; Tauer, L W

    2007-10-01

    The profitability of feeding rumen-protected Met (RPMet) sources to produce milk protein was estimated using a 2-step procedure: First, the effect of Met in metabolizable protein (MP) on milk protein production was estimated by using a quadratic Box-Cox functional form. Then, using these estimation results, the amounts of RPMet supplement that corresponded to the optimal levels of Met in MP for maximizing milk protein production and profit on dairy farms were determined. The data used in this study were modified from data used to determine the optimal level of Met in MP for lactating cows in the Nutrient Requirements of Dairy Cattle (NRC, 2001). The data used in this study differ from that in the NRC (2001) data in 2 ways. First, because dairy feed generally contains 1.80 to 1.90% Met in MP, this study adjusts the reference production value (RPV) from 2.06 to 1.80 or 1.90%. Consequently, the milk protein production response is also modified to an RPV of 1.80 or 1.90% Met in MP. Second, because this study is especially interested in how much additional Met, beyond the 1.80 or 1.90% already contained in the basal diet, is required to maximize farm profits, the data used are limited to concentrations of Met in MP above 1.80 or 1.90%. This allowed us to calculate any additional cost to farmers based solely on the price of an RPMet supplement and eliminated the need to estimate the dollar value of each gram of Met already contained in the basal diet. Results indicated that the optimal level of Met in MP for maximizing milk protein production was 2.40 and 2.42%, where the RPV was 1.80 and 1.90%, respectively. These optimal levels were almost identical to the recommended level of Met in MP of 2.40% in the NRC (2001). The amounts of RPMet required to increase the percentage of Met in MP from each RPV to 2.40 and 2.42% were 21.6 and 18.5 g/d, respectively. On the other hand, the optimal levels of Met in MP for maximizing profit were 2.32 and 2.34%, respectively. The amounts

  4. PII Signal Transduction Proteins, Pivotal Players in Microbial Nitrogen Control

    PubMed Central

    Arcondéguy, Tania; Jack, Rachael; Merrick, Mike

    2001-01-01

    The PII family of signal transduction proteins are among the most widely distributed signal proteins in the bacterial world. First identified in 1969 as a component of the glutamine synthetase regulatory apparatus, PII proteins have since been recognized as playing a pivotal role in control of prokaryotic nitrogen metabolism. More recently, members of the family have been found in higher plants, where they also potentially play a role in nitrogen control. The PII proteins can function in the regulation of both gene transcription, by modulating the activity of regulatory proteins, and the catalytic activity of enzymes involved in nitrogen metabolism. There is also emerging evidence that they may regulate the activity of proteins required for transport of nitrogen compounds into the cell. In this review we discuss the history of the PII proteins, their structures and biochemistry, and their distribution and functions in prokaryotes. We survey data emerging from bacterial genome sequences and consider other likely or potential targets for control by PII proteins. PMID:11238986

  5. Identification of a Class of Protein ADP-Ribosylating Sirtuins in Microbial Pathogens

    PubMed Central

    Rack, Johannes Gregor Matthias; Morra, Rosa; Barkauskaite, Eva; Kraehenbuehl, Rolf; Ariza, Antonio; Qu, Yue; Ortmayer, Mary; Leidecker, Orsolya; Cameron, David R.; Matic, Ivan; Peleg, Anton Y.; Leys, David; Traven, Ana; Ahel, Ivan

    2015-01-01

    Summary Sirtuins are an ancient family of NAD+-dependent deacylases connected with the regulation of fundamental cellular processes including metabolic homeostasis and genome integrity. We show the existence of a hitherto unrecognized class of sirtuins, found predominantly in microbial pathogens. In contrast to earlier described classes, these sirtuins exhibit robust protein ADP-ribosylation activity. In our model organisms, Staphylococcus aureus and Streptococcus pyogenes, the activity is dependent on prior lipoylation of the target protein and can be reversed by a sirtuin-associated macrodomain protein. Together, our data describe a sirtuin-dependent reversible protein ADP-ribosylation system and establish a crosstalk between lipoylation and mono-ADP-ribosylation. We propose that these posttranslational modifications modulate microbial virulence by regulating the response to host-derived reactive oxygen species. PMID:26166706

  6. Structures and functions of autotransporter proteins in microbial pathogens.

    PubMed

    Benz, Inga; Schmidt, M Alexander

    2011-08-01

    Since their discovery more than 20 years ago the autotransporter protein superfamily has been growing continuously and currently represents the largest protein family in (pathogenic) Gram-negative bacteria. Autotransporter proteins (AT) adhere to a common structural principle and are composed of a C-terminal β-barrel-shaped 'translocator' domain and an N-terminal 'passenger' domain. The translocator is anchored in the outer membrane and is indispensable for the N-terminal passenger part to traverse the outer membrane. Most if not all AT harbor a chaperone segment that increases protein stability and may be located in the passenger or translocator domain. The passenger mediates the specific virulence function(s) of the particular AT. Accordingly, passenger domains of AT can be quite variable. Interestingly, AT have been identified as the first glycosylated proteins in Gram-negative bacteria. Despite the considerable efforts invested in the characterization of autotransporter biogenesis, various aspects such as the participation of accessory proteins, the fate of the translocator, or the translocation of glycosylated proteins still remain only poorly understood. In addition, recent evidence indicates that the prefix 'auto' might be slightly exaggerated. Here, we will selectively discuss novel insights at various stages of AT biogenesis. PMID:21616712

  7. Supplementation of Flemingia macrophylla and cassava foliage as a rumen enhancer on fermentation efficiency and estimated methane production in dairy steers.

    PubMed

    Phesatcha, Burarat; Wanapat, Metha; Phesatcha, Kampanat; Ampapon, Thiwakorn; Kang, Sungchhang

    2016-10-01

    Four rumen-fistulated dairy steers, 3 years old with 180 ± 15 kg body weight (BW), were randomly assigned according to a 4 × 4 Latin square design to investigate on the effect of Flemingia macrophylla hay meal (FMH) and cassava hay meal (CH) supplementation on rumen fermentation efficiency and estimated methane production. The treatments were as follows: T1 = non-supplement, T2 = CH supplementation at 150 g/head/day, T3 = FMH supplementation at 150 g/head/day, and T4 = CH + FMH supplementation at 75 and 75 g/head/day. All steers were fed rice straw ad libitum and concentrate was offered at 0.5 % of BW. Results revealed that supplementation of CH and/or FMH did not affect on feed intake (P > 0.05) while digestibility of crude protein and neutral detergent fiber were increased especially in steers receiving FMH and CH+FMH (P < 0.05). Ruminal pH, temperature, and blood urea nitrogen were similar among treatments while ammonia nitrogen was increased in all supplemented groups (P < 0.05). Furthermore, propionic acid (C3) was increased while acetic acid (C2), C2:C3 ratio, and estimated methane production were decreased by dietary treatments. Protozoa and fungi population were not affected by dietary supplement while viable bacteria count increased in steers receiving FMH. Supplementation of FMH and/or FMH+CH increased microbial crude protein and efficiency of microbial nitrogen supply. This study concluded FMH (150 g/head/day) and/or CH+FMH (75 and 75 g/head/day) supplementation could be used as a rumen enhancer for increasing nutrient digestibility, rumen fermentation efficiency, and microbial protein synthesis while decreasing estimated methane production without adverse effect on voluntary feed intake of dairy steers fed rice straw.

  8. Metal affinity enrichment increases the range and depth of proteome identification for extracellular microbial proteins

    SciTech Connect

    Wheeler, Korin; Erickson, Brian K; Mueller, Ryan; Singer, Steven; Verberkmoes, Nathan C; Hwang, Mona; Thelen, Michael P.; Hettich, Robert {Bob} L

    2012-01-01

    Many key proteins, such as those involved in cellular signaling or transcription, are difficult to measure in microbial proteomic experiments due to the interfering presence of more abundant, dominant proteins. In an effort to enhance the identification of previously undetected proteins, as well as provide a methodology for selective enrichment, we evaluated and optimized immobilized metal affinity chromatography (IMAC) coupled with mass spectrometric characterization of extracellular proteins from an extremophilic microbial community. Seven different metals were tested for IMAC enrichment. The combined results added 20% greater proteomic depth to the extracellular proteome. Although this IMAC enrichment could not be conducted at the physiological pH of the environmental system, this approach did yield a reproducible and specific enrichment of groups of proteins with functions potentially vital to the community, thereby providing a more extensive biochemical characterization. Notably, 40 unknown proteins previously annotated as hypothetical were enriched and identified for the first time. Examples of identified proteins includes a predicted TonB signal sensing protein homologous to other known TonB proteins and a protein with a COXG domain previously identified in many chemolithoautotrophic microbes as having a function in the oxidation of CO.

  9. Microbial response to single-cell protein production and brewery wastewater treatment.

    PubMed

    Lee, Jackson Z; Logan, Andrew; Terry, Seth; Spear, John R

    2015-01-01

    As global fisheries decline, microbial single-cell protein (SCP) produced from brewery process water has been highlighted as a potential source of protein for sustainable animal feed. However, biotechnological investigation of SCP is difficult because of the natural variation and complexity of microbial ecology in wastewater bioreactors. In this study, we investigate microbial response across a full-scale brewery wastewater treatment plant and a parallel pilot bioreactor modified to produce an SCP product. A pyrosequencing survey of the brewery treatment plant showed that each unit process selected for a unique microbial community. Notably, flow equalization basins were dominated by Prevotella, methanogenesis effluent had the highest levels of diversity, and clarifier wet-well samples were sources of sequences for the candidate bacterial phyla of TM7 and BD1-5. Next, the microbial response of a pilot bioreactor producing SCP was tracked over 1 year, showing that two different production trials produced two different communities originating from the same starting influent. However, SCP production resulted generally in enrichment of several clades of rhizospheric diazotrophs of Alphaproteobacteria and Betaproteobacteria in the bioreactor and even more so in the final product. These diazotrophs are potentially useful as the basis of a SCP product for commercial feed production. PMID:24837420

  10. Microbial response to single-cell protein production and brewery wastewater treatment

    PubMed Central

    Lee, Jackson Z; Logan, Andrew; Terry, Seth; Spear, John R

    2015-01-01

    As global fisheries decline, microbial single-cell protein (SCP) produced from brewery process water has been highlighted as a potential source of protein for sustainable animal feed. However, biotechnological investigation of SCP is difficult because of the natural variation and complexity of microbial ecology in wastewater bioreactors. In this study, we investigate microbial response across a full-scale brewery wastewater treatment plant and a parallel pilot bioreactor modified to produce an SCP product. A pyrosequencing survey of the brewery treatment plant showed that each unit process selected for a unique microbial community. Notably, flow equalization basins were dominated by Prevotella, methanogenesis effluent had the highest levels of diversity, and clarifier wet-well samples were sources of sequences for the candidate bacterial phyla of TM7 and BD1-5. Next, the microbial response of a pilot bioreactor producing SCP was tracked over 1 year, showing that two different production trials produced two different communities originating from the same starting influent. However, SCP production resulted generally in enrichment of several clades of rhizospheric diazotrophs of Alphaproteobacteria and Betaproteobacteria in the bioreactor and even more so in the final product. These diazotrophs are potentially useful as the basis of a SCP product for commercial feed production. PMID:24837420

  11. Effect of bromochloromethane and fumarate on phylogenetic diversity of the formyltetrahydrofolate synthetase gene in bovine rumen.

    PubMed

    Mitsumori, Makoto; Matsui, Hiroki; Tajima, Kiyoshi; Shinkai, Takumi; Takenaka, Akio; Denman, Stuart E; McSweeney, Christopher S

    2014-01-01

    Effect of the methane inhibitor, bromochloromethane (BCM) and dietary substrate, fumarate, on microbial community structure of acetogen bacteria in the bovine rumen was investigated through analysis of the formyltetrahydrofolate synthetase gene (fhs). The fhs sequences obtained from BCM-untreated, BCM-treated, fumarate-untreated and fumarate-treated bovine rumen were categorized into homoacetogens and nonhomoacetogenic bacteria by homoacetogen similarity scores. Phylogenetic tree analysis indicated that most of the fhs sequences categorized into homoacetogens were divided into nine clusters, which were in close agreement with a result shown in a self-organizing map. The diversity of the fhs sequences from the BCM-treated rumen was significantly different from those from BCM-non-treated rumen. Principal component analysis also showed that addition of BCM to the rumen altered the population structure of acetogenic bacteria significantly but the effect of fumarate was comparatively minor. These results indicate that BCM affects diversity of actogens in the bovine rumen, and changes in acetogenic community structure in response to methane inhibitors may be caused by different mechanisms.

  12. Effects and mode of action of chitosan and ivy fruit saponins on the microbiome, fermentation and methanogenesis in the rumen simulation technique.

    PubMed

    Belanche, Alejandro; Pinloche, Eric; Preskett, David; Newbold, C Jamie

    2016-01-01

    This study investigates the effects of supplementing a control diet (CON) with chitosan (CHI) or ivy fruit saponins (IVY) as natural feed additives. Both additives had similar abilities to decrease rumen methanogenesis (-42% and -40%, respectively) using different mechanisms: due to its antimicrobial and nutritional properties CHI promoted a shift in the fermentation pattern towards propionate production which explained about two thirds of the decrease in methanogenesis. This shift was achieved by a simplification of the structure in the bacterial community and a substitution of fibrolytic (Firmicutes and Fibrobacteres) by amylolytic bacteria (Bacteroidetes and Proteobacteria) which led to greater amylase activity, lactate and microbial protein yield with no detrimental effect on feed digestibility. Contrarily, IVY had negligible nutritional properties promoting minor changes in the fermentation pattern and on the bacterial community. Instead, IVY modified the structure of the methanogen community and decreased its diversity. This specific antimicrobial effect of IVY against methanogens was considered its main antimethanogenic mechanism. IVY had however a negative impact on microbial protein synthesis. Therefore, CHI and IVY should be further investigated in vivo to determine the optimum doses which maintain low methanogenesis but prevent negative effects on the rumen fermentation and animal metabolism. PMID:26676056

  13. Transcriptome differences in the rumen of beef steers with variation in feed intake and gain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Feed efficiency is an economically important trait in beef production. The rumen wall interacts with feed, microbial populations and volatile fatty acids important to ruminant nutrition indicating it may play a critical role in the beef steer’s ability to utilize feedstuffs efficiently. To identif...

  14. Changes in the rumen microbiome from steers differing in feed efficiency

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle rumen has a diverse microbial ecosystem that is essential to allow the host to digest plant material. Changes in the composition and diversity of the ruminal microbiota have been associated with diet and age. Extremes in body weight (BW) gain in mice and humans have been associated with...

  15. Next generation sequencing to define prokaryotic and fungal diversity in the bovine rumen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A combination of Sanger and 454 sequences of small subunit rRNA loci were used to interrogate the microbial diversity in the bovine rumen of 14 pasture-fed animals. The observed bacterial species richness, based on the V1-V3 region of the 15S rRNA gene, was between 1902 to 2596 species-level operati...

  16. Changes in fatty acid composition of various full fat crushed oilseeds and their free oils when incubated with rumen liquor in vitro.

    PubMed

    Hoffmann, Achim; Steingass, Herbert; Schollenberger, Margit; Jara, Helga Terry; Hartung, Karin; Weiss, Eva; Mosenthin, Rainer

    2013-02-01

    The fatty acid pattern of dietary lipids can be modified during rumen biohydrogenation (BH). The objective of the present study was to assess changes in the FA pattern of different oilseed products supplied either as crushed full fat oilseed or as free oil after in vitro incubation with buffered rumen liquor. The FA patterns were determined at the beginning and compared with those measured after 24 h of incubation. The contents of fatty acids (FA) < C18 increased (p < 0.05) in nearly all treatments, eventually due to microbial de novo synthesis and fermentation of carbohydrates and proteins during incubation. In contrast, the contents of the dominating C18 FA, (oleic acid - C18:1c9, linoleic acid - C18:2c9,12, linolenic acid - C18:3c9,12,15) were reduced due to BH, resulting in the accumulation of characteristic BH intermediates, such as conjugated linoleic acid (CLA) isomer C18:2c9t11 (rumenic acid). However, both for crushed full fat oilseeds and their free oils the process of BH was not completed at the end of incubation. The disappearance was highest for C18:3c9,12,15, followed by C18:2c9,12 and C18:1c9. The rate of BH of unsaturated FA was higher in the crushed form compared to the oil form. Higher amounts of BH intermediates accumulated in the crushed form. Obviously, the physical form affects the degree of BH in vitro. The current results suggest that feeding crushed full fat seeds instead of their free oils to dairy cows might stimulate the formation of beneficial BH intermediates such as CLA in the rumen.

  17. Downregulation of Cellular Protective Factors of Rumen Epithelium in Goats Fed High Energy Diet

    PubMed Central

    Hollmann, Manfred; Miller, Ingrid; Hummel, Karin; Sabitzer, Sonja; Metzler-Zebeli, Barbara U.; Razzazi-Fazeli, Ebrahim; Zebeli, Qendrim

    2013-01-01

    Energy-rich diets can challenge metabolic and protective functions of the rumen epithelial cells, but the underlying factors are unclear. This study sought to evaluate proteomic changes of the rumen epithelium in goats fed a low, medium, or high energy diet. Expression of protein changes were compared by two-dimensional differential gel electrophoresis followed by protein identification with matrix assisted laser desorption ionisation tandem time-of-flight mass spectrometry. Of about 2,000 spots commonly detected in all gels, 64 spots were significantly regulated, which were traced back to 24 unique proteins. Interestingly, the expression profiles of several chaperone proteins with important cellular protective functions such as heat shock cognate 71 kDa protein, peroxiredoxin-6, serpin H1, protein disulfide-isomerase, and selenium-binding protein were collectively downregulated in response to high dietary energy supply. Similar regulation patterns were obtained for some other proteins involved in transport or metabolic functions. In contrast, metabolic enzymes like retinal dehydrogenase 1 and ATP synthase subunit beta, mitochondrial precursor were upregulated in response to high energy diet. Lower expressions of chaperone proteins in the rumen epithelial cells in response to high energy supply may suggest that these cells were less protected against the potentially harmful rumen toxic compounds, which might have consequences for rumen and systemic health. Our findings also suggest that energy-rich diets and the resulting acidotic insult may render rumen epithelial cells more vulnerable to cellular damage by attenuating their cell defense system, hence facilitating the impairment of rumen barrier function, typically observed in energy-rich fed ruminants. PMID:24349094

  18. Utilization of digital differential display to identify differentially expressed genes related to rumen development.

    PubMed

    Kato, Daichi; Suzuki, Yutaka; Haga, Satoshi; So, KyoungHa; Yamauchi, Eri; Nakano, Miwa; Ishizaki, Hiroshi; Choi, Kichoon; Katoh, Kazuo; Roh, Sang-Gun

    2016-04-01

    This study aimed to identify the genes associated with the development of the rumen epithelium by screening for candidate genes by digital differential display (DDD) in silico. Using DDD in NCBI's UniGene database, expressed sequence tag (EST)-based gene expression profiles were analyzed in rumen, reticulum, omasum, abomasum and other tissues in cattle. One hundred and ten candidate genes with high expression in the rumen were derived from a library of all tissues. The expression levels of 11 genes in all candidate genes were analyzed in the rumen, reticulum, omasum and abomasum of nine Japanese Black male calves (5-week-old pre-weaning: n = 3; 15-week-old weaned calves: n = 6). Among the 11 genes, only 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), aldo-keto reductase family 1, member C1-like (AKR1C1), and fatty acid binding protein 3 (FABP3) showed significant changes in the levels of gene expression in the rumen between the pre- and post-weaning of calves. These results indicate that DDD analysis in silico can be useful for screening candidate genes related to rumen development, and that the changes in expression levels of three genes in the rumen may have been caused by weaning, aging or both.

  19. Effects of a new microbial α-amylase inhibitor protein on Helicoverpa armigera larvae.

    PubMed

    Zeng, Fanrong; Wang, Xiaojing; Cui, Jinjie; Ma, Yan; Li, Qiannan

    2013-03-01

    A new microbial α-amylase inhibitor gene was cloned and characterized. The encoded, recombinant, α-amylase inhibitor protein was induced and expressed by isopropyl β-d-1-thiogalactopyranoside (IPTG) in Escherichia coli M15 cells. The effects of the α-amylase inhibitor protein on Helicoverpa armigera larvae were studied. Compared to the control, the weight of H. armigera larvae fed the diet with recombinant α-amylase inhibitor protein added at a concentration of 20 μg/g was reduced by 49.8%. The total soluble protein of H. armigera larvae fed the diet with the α-amylase inhibitor protein added was also reduced by 36.8% compared to the control. The recombinant α-amylase inhibitor protein showed inhibition activity against α-amylase of H. armigera. These results suggested that this α-amylase inhibitor protein may be a promising bioinsecticide candidate for controlling H. armigera.

  20. Effects of dietary supplementation of rumen-protected folic acid on rumen fermentation, degradability and excretion of urinary purine derivatives in growing steers.

    PubMed

    Wang, Cong; Liu, Qiang; Guo, Gang; Huo, WenJie; Ma, Le; Zhang, YanLi; Pei, CaiXia; Zhang, ShuanLin; Wang, Hao

    2016-12-01

    The present experiment was undertaken to determine the effects of dietary addition of rumen-protected folic acid (RPFA) on ruminal fermentation, nutrient degradability, enzyme activity and the relative quantity of ruminal cellulolytic bacteria in growing beef steers. Eight rumen-cannulated Jinnan beef steers averaging 2.5 years of age and 419 ± 1.9 kg body weight were used in a replicated 4 × 4 Latin square design. The four treatments comprised supplementation levels of 0 (Control), 70, 140 and 210 mg RPFA/kg dietary dry matter (DM). On DM basis, the ration consisted of 50% corn silage, 47% concentrate and 3% soybean oil. The DM intake (averaged 8.5 kg/d) was restricted to 95% of ad libitum intake. The intake of DM, crude protein (CP) and net energy for growth was not affected by treatments. In contrast, increasing RPFA supplementation increased average daily gain and the concentration of total volatile fatty acid and reduced ruminal pH linearly. Furthermore, increasing RPFA supplementation enhanced the acetate to propionate ratio and reduced the ruminal ammonia N content linearly. The ruminal effective degradability of neutral detergent fibre from corn silage and CP from concentrate improved linearly and was highest for the highest supplementation levels. The activities of cellobiase, xylanase, pectinase and α-amylase linearly increased, but carboxymethyl-cellulase and protease were not affected by the addition of RPFA. The relative quantities of Butyrivibrio fibrisolvens, Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes increased linearly. With increasing RPFA supplementation levels, the excretion of urinary purine derivatives was also increased linearly. The present results indicated that the supplementation of RPFA improved ruminal fermentation, nutrient degradability, activities of microbial enzymes and the relative quantity of the ruminal cellulolytic bacteria in a dose-dependent manner. According to the conditions of this

  1. Rendered-protein hydrolysates for microbial synthesis of cyanophycin biopolymer

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cyanophycin is a poly(arginyl-aspartate) biopolymer produced and stored intracellularly by bacteria. Cyanophycin has been proposed as a renewable replacement for petrochemical-based industrial products. An abundant source of amino acids and nitrogen such as in the form of protein hydrolysates is n...

  2. Evaluation of isoquinoline alkaloid supplementation levels on ruminal fermentation, characteristics of digestion, and microbial protein synthesis in steers fed a high-energy diet.

    PubMed

    Aguilar-Hernández, J A; Urías-Estrada, J D; López-Soto, M A; Barreras, A; Plascencia, A; Montaño, M; González-Vizcarra, V M; Estrada-Angulo, A; Castro-Pérez, B I; Barajas, R; Rogge, H I; Zinn, R A

    2016-01-01

    Four Holstein steers with ruminal and duodenal cannulas were used in a 4 × 4 Latin square design to examine the effect of daily intake of 0, 2, 4 or 6 g/steer of standardized plant extract containing a mixture of quaternary benzophenanthridine alkaloids and protopine alkaloids (QBA+PA) on the characteristics of ruminal fermentation and characteristics of digestion. The basal diet consisted of a steam-flaked corn-based finishing diet that contained 62% corn and 12% sudangrass hay and the rest of diet was composed of mainly dried distillers grains, molasses, fat, and minerals. The source of QBA+PA used was Sangrovit-RS (Phytobiotics Futterzusatzstoffe GmbH, Eltville, Germany) and supplementation levels of 2, 4, and 6 g Sangrovit-RS∙steer∙d, which represented a net daily ingestion of approximately 6, 12, and 18 mg of QBA+PA compounds, respectively. Inclusion of QBA+PA linearly increased ( = 0.04) flow to the duodenum of nonammonia N and linearly decreased ( < 0.01) duodenal flows of ammonia N. Ruminal microbial efficiency (duodenal microbial N; g/kg OM fermented in the rumen) and protein efficiency (duodenal nonammonia N; g/g N intake) were increased ( < 0.05) as the level of QBA+PA increased. There were no effects of QBA+PA supplementation on ruminal, postruminal, and total tract digestion of OM, starch, and NDF, but postruminal and total tract digestion of N increased ( < 0.01) as the level of QBA+PA increased. Digestible energy of the diet tended to increase (linear affect, = 0.09) with QBA+PA supplementation. Ruminal pH and total VFA molar concentrations were not different between treatments. Ruminal NH-N concentration linearly decreased ( = 0.02) with QBA+PA supplementation. Ruminal molar proportion of acetate increased ( = 0.04) as the supplementation level of QBA+PA increased. It is concluded that QBA+PA supplementation enhances efficiency of N utilization in feedlot steers fed a steam-flaked corn-based finishing diet. This effect was due, in part, to

  3. The effects of additives in napier grass silages on chemical composition, feed intake, nutrient digestibility and rumen fermentation.

    PubMed

    Bureenok, Smerjai; Yuangklang, Chalermpon; Vasupen, Kraisit; Schonewille, J Thomas; Kawamoto, Yasuhiro

    2012-09-01

    The effect of silage additives on ensiling characteristics and nutritive value of Napier grass (Pennisetum purpureum) silages was studied. Napier grass silages were made with no additive, fermented juice of epiphytic lactic acid bacteria (FJLB), molasses or cassava meal. The ensiling characteristics were determined by ensiling Napier grass silages in airtight plastic pouches for 2, 4, 7, 14, 21 and 45 d. The effect of Napier grass silages treated with these additives on voluntary feed intake, digestibility, rumen fermentation and microbial rumen fermentation was determined in 4 fistulated cows using 4×4 Latin square design. The pH value of the treated silages rapidly decreased, and reached to the lowest value within 7 d of the start of fermentation, as compared to the control. Lactic acid content of silages treated with FJLB was stable at 14 d of fermentation and constant until 45 d of ensiling. At 45 d of ensiling, neutral detergent fiber (NDF) and acid detergent fiber (ADF) of silage treated with cassava meal were significantly lower (p<0.05) than the others. In the feeding trial, the intake of silage increased (p<0.05) in the cow fed with the treated silage. Among the treatments, dry matter intake was the lowest in the silage treated with cassava meal. The organic matter, crude protein and NDF digestibility of the silage treated with molasses was higher than the silage without additive and the silage treated with FJLB. The rumen parameters: ruminal pH, ammonia-nitrogen (NH3-N), volatile fatty acid (VFA), blood urea nitrogen (BUN) and bacterial populations were not significantly different among the treatments. In conclusion, these studies confirmed that the applying of molasses improved fermentative quality, feed intake and digestibility of Napier grass.

  4. Manipulating ruminal fermentation: a microbial ecological perspective.

    PubMed

    Weimer, P J

    1998-12-01

    The essential role of ruminal microflora in ruminant nutrition provides the potential for improvement in animal production via altering the numbers or activities of specific classes of microorganisms. Successful alterations will be facilitated by an understanding of the microbial ecology of the rumen based on its mechanistic underpinnings. Demonstrated improvements in ruminal fermentation can be traced to their consonance with well-established principles of microbial ecology (niche occupancy, selective pressure, adaptation, and interactions) and the thermodynamics and kinetics of substrate utilization. Application of these principles to several proposed alterations of the ruminal bacterial population allows a prediction of their relative feasibility. Improving fiber digestion, decreasing protein degradation, and detoxifying feed components that are present in low concentrations will be difficult to achieve in the rumen and are best approached by altering the feed, either genetically or with postharvest treatment. By contrast, the detoxification of feed components present in high concentration, and redirection of electron disposal away from methanogenesis, are more productive targets for microbiological research.

  5. Thermally activated charge transport in microbial protein nanowires

    PubMed Central

    Lampa-Pastirk, Sanela; Veazey, Joshua P.; Walsh, Kathleen A.; Feliciano, Gustavo T.; Steidl, Rebecca J.; Tessmer, Stuart H.; Reguera, Gemma

    2016-01-01

    The bacterium Geobacter sulfurreducens requires the expression of conductive protein filaments or pili to respire extracellular electron acceptors such as iron oxides and uranium and to wire electroactive biofilms, but the contribution of the protein fiber to charge transport has remained elusive. Here we demonstrate efficient long-range charge transport along individual pili purified free of metal and redox organic cofactors at rates high enough to satisfy the respiratory rates of the cell. Carrier characteristics were within the orders reported for organic semiconductors (mobility) and inorganic nanowires (concentration), and resistivity was within the lower ranges reported for moderately doped silicon nanowires. However, the pilus conductance and the carrier mobility decreased when one of the tyrosines of the predicted axial multistep hopping path was replaced with an alanine. Furthermore, low temperature scanning tunneling microscopy demonstrated the thermal dependence of the differential conductance at the low voltages that operate in biological systems. The results thus provide evidence for thermally activated multistep hopping as the mechanism that allows Geobacter pili to function as protein nanowires between the cell and extracellular electron acceptors. PMID:27009596

  6. Thermally activated charge transport in microbial protein nanowires.

    PubMed

    Lampa-Pastirk, Sanela; Veazey, Joshua P; Walsh, Kathleen A; Feliciano, Gustavo T; Steidl, Rebecca J; Tessmer, Stuart H; Reguera, Gemma

    2016-01-01

    The bacterium Geobacter sulfurreducens requires the expression of conductive protein filaments or pili to respire extracellular electron acceptors such as iron oxides and uranium and to wire electroactive biofilms, but the contribution of the protein fiber to charge transport has remained elusive. Here we demonstrate efficient long-range charge transport along individual pili purified free of metal and redox organic cofactors at rates high enough to satisfy the respiratory rates of the cell. Carrier characteristics were within the orders reported for organic semiconductors (mobility) and inorganic nanowires (concentration), and resistivity was within the lower ranges reported for moderately doped silicon nanowires. However, the pilus conductance and the carrier mobility decreased when one of the tyrosines of the predicted axial multistep hopping path was replaced with an alanine. Furthermore, low temperature scanning tunneling microscopy demonstrated the thermal dependence of the differential conductance at the low voltages that operate in biological systems. The results thus provide evidence for thermally activated multistep hopping as the mechanism that allows Geobacter pili to function as protein nanowires between the cell and extracellular electron acceptors. PMID:27009596

  7. Rumen bacterial communities shift across a lactation in Holstein, Jersey and Holstein × Jersey dairy cows and correlate to rumen function, bacterial fatty acid composition and production parameters.

    PubMed

    Bainbridge, Melissa L; Cersosimo, Laura M; Wright, André-Denis G; Kraft, Jana

    2016-05-01

    Rumen bacteria form a dynamic, complex, symbiotic relationship with their host, degrading forages to provide volatile fatty acids (VFA) and other substrates as energy to the animal. The objectives were to characterize rumen bacteria in three genetic lines of primiparous dairy cattle, Holstein (HO, n = 7), Jersey (JE, n = 8), and HO × JE crossbreeds (CB, n = 7) across a lactation [3, 93, 183 and 273 days in milk (DIM)] and correlate these factors with VFA, bacterial cell membrane fatty acids (FA), and animal production (i.e. milk yield). This study employed Illumina MiSeq (v. 3) to investigate rumen bacterial communities and gas-liquid chromatography/mass spectroscopy to identify bacterial membrane FA. Lactation stage had a prominent effect on rumen bacterial communities, whereas genetics had a lesser effect on rumen bacteria. The FA composition of bacterial cell membranes was affected by both lactation stage and genetics. Few correlations existed between VFA and bacterial communities; however, moderate correlations occurred between milk yield, protein percentage, fat yield and rumen bacterial communities. Positive correlations were found between branched-chain FA (BCFA) in bacterial cell membranes and bacterial genera. In conclusion, bacterial communities and their FA compositions are more affected by stage of lactation than by genetics of dairy cow. PMID:26985012

  8. Impact of Enzymatic and Microbial Bioprocessing on Protein Modification and Nutritional Properties of Wheat Bran.

    PubMed

    Arte, Elisa; Rizzello, Carlo G; Verni, Michela; Nordlund, Emilia; Katina, Kati; Coda, Rossana

    2015-10-01

    Besides providing dietary fiber, wheat bran is a recognized source of protein and is considered a very valuable substitute for other protein-rich sources in the food and feed industry. Nonetheless, several factors affect protein bioavailability, including bran's layered structure. This study showed the influence on the release and protein modification of wheat bran of different bioprocessing methods involving the activation of endogenous enzymes of bran, the addition of an enzyme mixture having carbohydrase activity, and microbial fermentation. Bioprocessing in acidic conditions significantly enhanced the solubilization of protein from wheat bran, reaching the highest value in the treatment where the sole endogenous protease activity was activated. Bioprocessing through controlled fermentation allowed a more intense proteolysis and strongly impacted the in vitro digestibility of proteins. The combined use of starter cultures and cell-wall-degrading enzymes was characterized by the highest increase of phytase activity and total phenols. PMID:26365885

  9. Impact of Enzymatic and Microbial Bioprocessing on Protein Modification and Nutritional Properties of Wheat Bran.

    PubMed

    Arte, Elisa; Rizzello, Carlo G; Verni, Michela; Nordlund, Emilia; Katina, Kati; Coda, Rossana

    2015-10-01

    Besides providing dietary fiber, wheat bran is a recognized source of protein and is considered a very valuable substitute for other protein-rich sources in the food and feed industry. Nonetheless, several factors affect protein bioavailability, including bran's layered structure. This study showed the influence on the release and protein modification of wheat bran of different bioprocessing methods involving the activation of endogenous enzymes of bran, the addition of an enzyme mixture having carbohydrase activity, and microbial fermentation. Bioprocessing in acidic conditions significantly enhanced the solubilization of protein from wheat bran, reaching the highest value in the treatment where the sole endogenous protease activity was activated. Bioprocessing through controlled fermentation allowed a more intense proteolysis and strongly impacted the in vitro digestibility of proteins. The combined use of starter cultures and cell-wall-degrading enzymes was characterized by the highest increase of phytase activity and total phenols.

  10. Meta-analysis of postruminal microbial nitrogen flows in dairy cattle. I. Derivation of equations.

    PubMed

    Roman-Garcia, Yairanex; White, Robin R; Firkins, Jeffrey L

    2016-10-01

    The objective was to summarize the literature and derive equations that relate the chemical composition of diet and rumen characteristics to the intestinal supply of microbial nitrogen (MicN), efficiency of microbial protein synthesis (EMPS), and flow of nonammonia nonmicrobial N (NANMN). In this study, 619 treatment means from 183 trials were assembled for dairy cattle sampled from the duodenum or omasum. Backward elimination multiple regression was used to derive equations to estimate flow of nitrogenous components over a large range of dietary conditions. An intercept shift for sample location revealed that omasal sampling estimated greater MicN flow relative to duodenal sampling, but sample location did not interact with any other variables tested. The ruminal outflow of MicN was positively associated with dry matter intake (DMI) and with dietary starch percentage at a decreasing rate (quadratic response). Also, MicN was associated with DMI and rumen-degraded starch and neutral detergent fiber (NDF). When rumen measurements were included, ruminal pH and ammonia-N were negatively related to MicN flow along with a strong positive association with ruminal isovalerate molar proportion. When evaluating these variables with EMPS, isovalerate interacted with ammonia such that the slope for EMPS with increasing isovalerate increased as ammonia-N concentration decreased. A similar equation with isobutyrate confirms the importance of branched-chain volatile fatty acids to increase growth rate and therefore assimilation of ammonia-N into microbial protein. The ruminal outflow of NANMN could be predicted by dietary NDF and crude protein percentages, which also interacted. This result is probably associated with neutral detergent insoluble N contamination of NDF in certain rumen-undegradable protein sources. Because NANMN is calculated by subtracting MicN, sample location was inversely related compared with the MicN equation, and omasal sampling underestimated NANMN

  11. PATtyFams: Protein Families for the Microbial Genomes in the PATRIC Database

    PubMed Central

    Davis, James J.; Gerdes, Svetlana; Olsen, Gary J.; Olson, Robert; Pusch, Gordon D.; Shukla, Maulik; Vonstein, Veronika; Wattam, Alice R.; Yoo, Hyunseung

    2016-01-01

    The ability to build accurate protein families is a fundamental operation in bioinformatics that influences comparative analyses, genome annotation, and metabolic modeling. For several years we have been maintaining protein families for all microbial genomes in the PATRIC database (Pathosystems Resource Integration Center, patricbrc.org) in order to drive many of the comparative analysis tools that are available through the PATRIC website. However, due to the burgeoning number of genomes, traditional approaches for generating protein families are becoming prohibitive. In this report, we describe a new approach for generating protein families, which we call PATtyFams. This method uses the k-mer-based function assignments available through RAST (Rapid Annotation using Subsystem Technology) to rapidly guide family formation, and then differentiates the function-based groups into families using a Markov Cluster algorithm (MCL). This new approach for generating protein families is rapid, scalable and has properties that are consistent with alignment-based methods. PMID:26903996

  12. Identification of Biofilm Matrix-Associated Proteins from an Acid Mine Drainage Microbial Community

    SciTech Connect

    Jiao, Yongqin; D'Haeseleer, Patrik M; Dill, Brian; Shah, Manesh B; Verberkmoes, Nathan C; Hettich, Robert {Bob} L; Banfield, Jillian F.; Thelen, Michael P.

    2011-01-01

    In microbial communities, extracellular polymeric substances (EPS), also called the extracellular matrix, provide the spatial organization and structural stability during biofilm development. One of the major components of EPS is protein, but it is not clear what specific functions these proteins contribute to the extracellular matrix or to microbial physiology. To investigate this in biofilms from an extremely acidic environment, we used shotgun proteomics analyses to identify proteins associated with EPS in biofilms at two developmental stages, designated DS1 and DS2. The proteome composition of the EPS was significantly different from that of the cell fraction, with more than 80% of the cellular proteins underrepresented or undetectable in EPS. In contrast, predicted periplasmic, outer membrane, and extracellular proteins were overrepresented by 3- to 7-fold in EPS. Also, EPS proteins were more basic by 2 pH units on average and about half the length. When categorized by predicted function, proteins involved in motility, defense, cell envelope, and unknown functions were enriched in EPS. Chaperones, such as histone-like DNA binding protein and cold shock protein, were overrepresented in EPS. Enzymes, such as protein peptidases, disulfide-isomerases, and those associated with cell wall and polysaccharide metabolism, were also detected. Two of these enzymes, identified as -N-acetylhexosaminidase and cellulase, were confirmed in the EPS fraction by enzymatic activity assays. Compared to the differences between EPS and cellular fractions, the relative differences in the EPS proteomes between DS1 and DS2 were smaller and consistent with expected physiological changes during biofilm development.

  13. Use of Adenosine 5′-Triphosphate as an Indicator of the Microbiota Biomass in Rumen Contents

    PubMed Central

    Forsberg, C. W.; Lam, K.

    1977-01-01

    A number of techniques were tested for their efficiency in extracting adenosine 5′-triphosphate (ATP) from strained rumen fluid (SRF). Extraction with 0.6 N H2SO4, using a modification of the procedure described by Lee et al. (1971), was the most efficient and was better suited for extracting particulate samples. Neutralized extracts could not be stored frozen before assaying for ATP because large losses were incurred. The inclusion of internal standards was necessary to correct for incomplete recovery of ATP. The ATP concentration in rumen contents from a cow receiving a ration of dried roughage (mainly alfalfa hay) ranged from 31 to 56 μg of ATP per g of contents. Approximately 75% of the ATP was associated with the particulate material. The ATP was primarily of microbial origin, since only traces of ATP were present in the feed and none was found in “cell-free” rumen fluid. Fractionation of the bacterial and protozoal populations in SRF resulted in the isolation of an enriched protozoal fraction with a 10-fold higher ATP concentration than that of the separated rumen bacteria. The ATP pool sizes of nine functionally important rumen bacteria during the exponential phase of growth ranged from 1.1 to 17.6 μg of ATP per mg of dry weight. This information indicates that using ATP as a measure of microbial biomass in rumen contents must be done with caution because of possible variations in the efficiency of extraction of ATP from rumen contents and differences in the concentration of ATP in rumen microbes. PMID:16345203

  14. Short communication: Initial evidence supporting existence of potential rumen epidermal stem and progenitor cells.

    PubMed

    Yohe, T T; Tucker, H L M; Parsons, C L M; Geiger, A J; Akers, R M; Daniels, K M

    2016-09-01

    The bovine rumen epidermis is a keratinized multilayered tissue that experiences persistent cell turnover. Because of this constant cell turnover, epidermal stem cells and their slightly more differentiated daughter cells, epidermal progenitor cells, must exist in the stratum basale of rumen epidermis. To date, these 2 epidermal cell populations and any unique cellular markers they may possess remain completely uncharacterized in the bovine rumen. An important first step in this new research area is the demonstration of the relative abundance and existence of markers for these cells in rumen tissue. A related second step is to document rumen epidermal proliferative responses to an extrinsic signal such as nutrient concentration within the rumen. The objectives of this experiment were to evaluate the extrinsic effect of diet on (1) gene expression of 6 potential rumen epidermal stem or progenitor cell markers and (2) rumen epidermal cell proliferation within the stratum basale. Twelve preweaned Holstein heifers were fed either a restricted diet (R) or an enhanced diet (EH). Animals on R received a milk replacer (MR) diet fed at 0.44kg of powder dry matter (DM)/d (20.9% crude protein, 29.8% fat, DM basis) and EH received MR at 1.08kg of powder dry matter/d (28.9% crude protein, 26.2% fat, DM basis). All calves had access to a 20% crude protein starter and were weaned during wk 7 of the experiment. Lifetime DM intake was 0.73kg of DM/calf per day for R (5.88 Mcal of net energy/calf per day) and 1.26kg of DM/calf per day for EH (10.68 Mcal of net energy/calf per day). Twenty-four hours before slaughter heifers received an intravenous dose of 5-bromo-2'-deoxyuridine to label proliferating cells. Heifers were slaughtered at 8 wk of age, and rumen samples from the ventral sac region were obtained and stored in RNA preservative and processed for routine histology. Quantitative real-time reverse transcriptase PCR was used to analyze relative abundance of genes. Candidate

  15. Use of Modern Chemical Protein Synthesis and Advanced Fluorescent Assay Techniques to Experimentally Validate the Functional Annotation of Microbial Genomes

    SciTech Connect

    Kent, Stephen

    2012-07-20

    The objective of this research program was to prototype methods for the chemical synthesis of predicted protein molecules in annotated microbial genomes. High throughput chemical methods were to be used to make large numbers of predicted proteins and protein domains, based on microbial genome sequences. Microscale chemical synthesis methods for the parallel preparation of peptide-thioester building blocks were developed; these peptide segments are used for the parallel chemical synthesis of proteins and protein domains. Ultimately, it is envisaged that these synthetic molecules would be ‘printed’ in spatially addressable arrays. The unique ability of total synthesis to precision label protein molecules with dyes and with chemical or biochemical ‘tags’ can be used to facilitate novel assay technologies adapted from state-of-the art single molecule fluorescence detection techniques. In the future, in conjunction with modern laboratory automation this integrated set of techniques will enable high throughput experimental validation of the functional annotation of microbial genomes.

  16. Characterization of 18:1 and 18:2 isomers produced during microbial biohydrogenation of unsaturated fatty acids from canola and soya bean oil in the rumen of lactating cows.

    PubMed

    Loor, J J; Bandara, A B P A; Herbein, J H

    2002-12-01

    Ruminal production of biohydrogenation intermediates in response to unsaturated oils was assessed using 24 Jersey cows fed a control diet or the control diet supplemented at 35 g/kg dry matter (DM) with canola, soya bean, or a mixture of equal amounts of canola plus soya bean oil for 4-weeks. Total fatty acid content averaged 63 or 35 g/kg DM for oil-supplemented diets or control. Oleic acid accounted for 6, 29, 21 or 12 g/kg DM in the control, canola, mixture, or soya bean oil diet, respectively. Linoleic acid averaged 17, 19, 26, or 33 g/kg DM and linolenic acid 5, 5, 6 or 8 g/kg DM for control, canola, mixture, or soya bean oil. Concentrations of cis12-, trans11-, trans13+14, and trans15-18:1 were 0.81, 2.99, 2.24, and 0.73 mg/g rumen fluid, respectively, in response to soya bean oil and were 126, 90, 45, and 38% greater compared with other diets. Trans11cis15-, cis9trans11- and cis9 cis11-18:2 also were greater when soya bean oil (0.30, 0.34 and 0.01 mg/g, respectively) was fed compared with other treatments (0.12, 0.21 and 0.004 mg/g, respectively). Feeding canola oil resulted in greater concentrations of trans4-, trans5-, trans6+7+8-, trans9- and trans10-18:1 (0.20, 0.25, 0.87, 0.39 and 0.70 mg/g, respectively) compared with other diets (0.09, 0.15, 0.36, 0.20 and 0.46 mg/g, respectively). Trans10cis12-18:2 concentration did not differ as a result of diet and averaged 0.002 mg/g rumen contents. The pattern of 18:1 and 18:2 isomers formed during ruminal biohydrogenation depends greatly on dietary profile of unsaturated fatty acids.

  17. In-depth diversity analysis of the bacterial community resident in the camel rumen.

    PubMed

    Gharechahi, Javad; Zahiri, Hossein Shahbani; Noghabi, Kambiz Akbari; Salekdeh, Ghasem Hosseini

    2015-02-01

    The rumen compartment of the ruminant digestive tract is an enlarged fermentation chamber which houses a diverse collection of symbiotic microorganisms that provide the host animal with a remarkable ability to digest plant lignocellulosic materials. Characterization of the ruminal microbial community provides opportunities to improve animal food digestion efficiency, mitigate methane emission, and develop efficient fermentation systems to convert plant biomasses into biofuels. In this study, 16S rRNA gene amplicon pyrosequencing was applied in order to explore the structure of the bacterial community inhabiting the camel rumen. Using 76,333 quality-checked, chimera- and singleton-filtered reads, 4954 operational taxonomic units (OTUs) were identified at a 97% species level sequence identity. At the phylum level, more than 96% of the reads were affiliated to OTUs belonging to Bacteroidetes (51%), Firmicutes (31%), Proteobacteria (4.8%), Spirochaetes (3.5%), Fibrobacteres (3.1%), Verrucomicrobia (2.7%), and Tenericutes (0.95%). A total of 15% of the OTUs (746) that contained representative sequences from all major taxa were shared by all animals and they were considered as candidate members of the core camel rumen microbiome. Analysis of microbial composition through the solid and liquid fractions of rumen digesta revealed differential enrichment of members of Fibrobacter, Clostridium, Ruminococcus, and Treponema in the solid fraction, as well as members of Prevotella, Verrucomicrobia, Cyanobacteria, and Succinivibrio in the liquid fraction. The results clearly showed that the camel rumen microbiome was structurally similar but compositionally distinct from that of other ruminants, such as the cow. The unique characteristic of the camel rumen microbiome that differentiated it from those of other ruminants was the significant enrichment for cellulolytic bacteria.

  18. Action of microbial transglutaminase (MTGase) in the modification of food proteins: a review.

    PubMed

    Gaspar, Ana Luisa Camolezi; de Góes-Favoni, Silvana Pedroso

    2015-03-15

    Microbial transglutaminase (MTGase) is an enzyme of the class of transferases widely known to modify protein functional properties in food systems. The main mechanisms of action involved are polymerisations, which result in changes in the molecule's hydrophobicity. Among the functional properties, MTGase affects solubility and hence gelation, emulsification, foaming, viscosity and water-holding capacity, which all depend on protein solubility. Although the enzyme's benefits for protein functionality in a variety of food systems are widely reported in the literature, few studies have focussed on its mechanisms. The purpose of this review is therefore to investigate the mechanisms of action of MTGase and consider its influence on the functional properties with different protein substrates. Understanding these mechanisms is the first step in the development and application, in food production, of new and better functional properties with improved and/or innovative characteristics that can satisfy consumer expectations.

  19. The effects of dietary sucrose and the concentration of plasma urea and rumen ammonia on the degradation of urea in the gastrointestinal tract of cattle.

    PubMed

    Kennedy, P M

    1980-01-01

    1. The rates of entry of urea into plasma, of urea degradation in the gastrointestinal tract, and the partition of that degradation between the rumen and post-ruminal tract were determined by use of [14C]urea and NaH14CO3 in Hereford steers receiving hay diets with or without sucrose. The concentrations of plasma urea and rumen ammonia were varied by infusions of urea into the rumen or abomasum. 2. For all diets, plasma urea concentration was related to urea entry rate, to degradation of urea in the whole gastrointestinal tract, and to its degradation in the post-ruminal tract, but the relationship with its degradation in the rumen was poor. 3. Degradation of urea in the rumen was related in a multiple regression in a curvilinear manner in three groups of diets (pasture-hay alone, pasture-hay--lucerne (Medicago sativa) mixtures, diets with sucrose), and negatively to rumen ammonia concentration for pasture-hay diets, and diets with sucrose. 4. Ruminal clearance of urea (rate of urea degradation per plasma urea concentration) was negatively related to the rumen ammonia concentration for steers given diets with sucrose, of pasture-hay with or without urea infusions. Provision of sucrose in the diet significantly increased clearance. 5. Enhanced urea degradation in the rumen associated with dietary sucrose supplements accounted for 0.4 of additional microbial N synthesis in the rumen. 6. The partition of transfer of urea to the rumen via saliva and through the rumen wall is discussed.

  20. Metagenomic assessment of the functional potential of the rumen microbiome in Holstein dairy cows.

    PubMed

    Pitta, Dipti W; Indugu, Nagaraju; Kumar, Sanjay; Vecchiarelli, Bonnie; Sinha, Rohini; Baker, Linda D; Bhukya, Bhima; Ferguson, James D

    2016-04-01

    The microbial ecology of the rumen microbiome is influenced by the diet and the physiological status of the dairy cow and can have tremendous influence on the yield and components of milk. There are significant differences in milk yields between first and subsequent lactations of dairy cows, but information on how the rumen microbiome changes as the dairy cow gets older has received little attention. We characterized the rumen microbiome of the dairy cow for phylogeny and functional pathways by lactation group and stage of lactation using a metagenomics approach. Our findings revealed that the rumen microbiome was dominated by Bacteroidetes (70%), Firmicutes (15-20%) and Proteobacteria (7%). The abundance of Firmicutes and Proteobacteria were independently influenced by diet and lactation. Bacteroidetes contributed to a majority of the metabolic functions in first lactation dairy cows while the contribution from Firmicutes and Proteobacteria increased incrementally in second and third lactation dairy cows. We found that nearly 70% of the CAZymes were oligosaccharide breaking enzymes which reflect the higher starch and fermentable sugars in the diet. The results of this study suggest that the rumen microbiome continues to evolve as the dairy cow advances in lactations and these changes may have a significant role in milk production.

  1. Metagenomic assessment of the functional potential of the rumen microbiome in Holstein dairy cows.

    PubMed

    Pitta, Dipti W; Indugu, Nagaraju; Kumar, Sanjay; Vecchiarelli, Bonnie; Sinha, Rohini; Baker, Linda D; Bhukya, Bhima; Ferguson, James D

    2016-04-01

    The microbial ecology of the rumen microbiome is influenced by the diet and the physiological status of the dairy cow and can have tremendous influence on the yield and components of milk. There are significant differences in milk yields between first and subsequent lactations of dairy cows, but information on how the rumen microbiome changes as the dairy cow gets older has received little attention. We characterized the rumen microbiome of the dairy cow for phylogeny and functional pathways by lactation group and stage of lactation using a metagenomics approach. Our findings revealed that the rumen microbiome was dominated by Bacteroidetes (70%), Firmicutes (15-20%) and Proteobacteria (7%). The abundance of Firmicutes and Proteobacteria were independently influenced by diet and lactation. Bacteroidetes contributed to a majority of the metabolic functions in first lactation dairy cows while the contribution from Firmicutes and Proteobacteria increased incrementally in second and third lactation dairy cows. We found that nearly 70% of the CAZymes were oligosaccharide breaking enzymes which reflect the higher starch and fermentable sugars in the diet. The results of this study suggest that the rumen microbiome continues to evolve as the dairy cow advances in lactations and these changes may have a significant role in milk production. PMID:26700882

  2. The effect of high polyphenol oxidase grass silage on metabolism of polyunsaturated fatty acids and nitrogen across the rumen of beef steers.

    PubMed

    Lee, M R F; Theobald, V J; Gordon, N; Leyland, M; Tweed, J K S; Fychan, R; Scollan, N D

    2014-11-01

    Polyphenol oxidase (PPO) activity in red clover (Trifolium pratense) has been reported to reduce both proteolysis and lipolysis, resulting in greater N use efficiency and protection of PUFA across the rumen. Although high levels of PPO have been reported in grasses such as cocksfoot (orchard grass; Dactylis glomerata), no in vivo research has determined whether grass PPO elicits the same response as red clover PPO. To test the hypothesis that silage ensiled from grass with high levels of PPO protects N and PUFA across the rumen, 6 steers with ruminal and duodenal cannulas were offered cocksfoot silage (CO; high-PPO grass), perennial ryegrass silage (PR; Lolium perenne; low-PPO grass), or red clover silage (RC; high-PPO control) at 16 g DM/kg BW daily with the experiment consisting of two 3 × 3 Latin squares with 21-d periods, consisting of 12 d of diet adaptation, 6 d of duodenal marker infusion, 2 d of duodenal sampling, and 1 d of ruminal sampling. All silages were well preserved, with DM of 34.4, 55.3, and 45.4% for CO, PR, and RC. Activity of PPO in silages was low due to deactivation but was greater in CO than either PR or RC (0.15 vs. 0.05 and 0.08 μkatal/g DM). Protein-bound phenol (mg/g DM) as a measure of the degree of oxidation and an indication of PPO protection was greatest for RC (15.9) but comparable for PR (10.1) and CO (12.2). Biohydrogenation of C18 PUFA was significantly lower on RC compared to the 2 grass silages with CO greater than PR. Despite lower levels of total fatty acid intake and subsequent duodenal flow, CO resulted in greater levels of phytanic acid and total branched and odd chain fatty acids in duodenal digesta than RC or PR. Ruminal ammonia concentration was greatest for RC, with no difference between the grasses. Duodenal flow of microbial N and efficiency of microbial protein synthesis were lowest for CO and comparable for RC and PR. The CO (high-grass PPO) did not result in elevated levels of C18 PUFA escaping the rumen or

  3. The effect of high polyphenol oxidase grass silage on metabolism of polyunsaturated fatty acids and nitrogen across the rumen of beef steers.

    PubMed

    Lee, M R F; Theobald, V J; Gordon, N; Leyland, M; Tweed, J K S; Fychan, R; Scollan, N D

    2014-11-01

    Polyphenol oxidase (PPO) activity in red clover (Trifolium pratense) has been reported to reduce both proteolysis and lipolysis, resulting in greater N use efficiency and protection of PUFA across the rumen. Although high levels of PPO have been reported in grasses such as cocksfoot (orchard grass; Dactylis glomerata), no in vivo research has determined whether grass PPO elicits the same response as red clover PPO. To test the hypothesis that silage ensiled from grass with high levels of PPO protects N and PUFA across the rumen, 6 steers with ruminal and duodenal cannulas were offered cocksfoot silage (CO; high-PPO grass), perennial ryegrass silage (PR; Lolium perenne; low-PPO grass), or red clover silage (RC; high-PPO control) at 16 g DM/kg BW daily with the experiment consisting of two 3 × 3 Latin squares with 21-d periods, consisting of 12 d of diet adaptation, 6 d of duodenal marker infusion, 2 d of duodenal sampling, and 1 d of ruminal sampling. All silages were well preserved, with DM of 34.4, 55.3, and 45.4% for CO, PR, and RC. Activity of PPO in silages was low due to deactivation but was greater in CO than either PR or RC (0.15 vs. 0.05 and 0.08 μkatal/g DM). Protein-bound phenol (mg/g DM) as a measure of the degree of oxidation and an indication of PPO protection was greatest for RC (15.9) but comparable for PR (10.1) and CO (12.2). Biohydrogenation of C18 PUFA was significantly lower on RC compared to the 2 grass silages with CO greater than PR. Despite lower levels of total fatty acid intake and subsequent duodenal flow, CO resulted in greater levels of phytanic acid and total branched and odd chain fatty acids in duodenal digesta than RC or PR. Ruminal ammonia concentration was greatest for RC, with no difference between the grasses. Duodenal flow of microbial N and efficiency of microbial protein synthesis were lowest for CO and comparable for RC and PR. The CO (high-grass PPO) did not result in elevated levels of C18 PUFA escaping the rumen or

  4. Leaf Treatments with a Protein-Based Resistance Inducer Partially Modify Phyllosphere Microbial Communities of Grapevine

    PubMed Central

    Cappelletti, Martina; Perazzolli, Michele; Antonielli, Livio; Nesler, Andrea; Torboli, Esmeralda; Bianchedi, Pier L.; Pindo, Massimo; Puopolo, Gerardo; Pertot, Ilaria

    2016-01-01

    Protein derivatives and carbohydrates can stimulate plant growth, increase stress tolerance, and activate plant defense mechanisms. However, these molecules can also act as a nutritional substrate for microbial communities living on the plant phyllosphere and possibly affect their biocontrol activity against pathogens. We investigated the mechanisms of action of a protein derivative (nutrient broth, NB) against grapevine downy mildew, specifically focusing on the effects of foliar treatments on plant defense stimulation and on the composition and biocontrol features of the phyllosphere microbial populations. NB reduced downy mildew symptoms and induced the expression of defense-related genes in greenhouse- and in vitro-grown plants, indicating the activation of grapevine resistance mechanisms. Furthermore, NB increased the number of culturable phyllosphere bacteria and altered the composition of bacterial and fungal populations on leaves of greenhouse-grown plants. Although, NB-induced changes on microbial populations were affected by the structure of indigenous communities originally residing on grapevine leaves, degrees of disease reduction and defense gene modulation were consistent among the experiments. Thus, modifications in the structure of phyllosphere populations caused by NB application could partially contribute to downy mildew control by competition for space or other biocontrol strategies. Particularly, changes in the abundance of phyllosphere microorganisms may provide a contribution to resistance induction, partially affecting the hormone-mediated signaling pathways involved. Modifying phyllosphere populations by increasing natural biocontrol agents with the application of selected nutritional factors can open new opportunities in terms of sustainable plant protection strategies. PMID:27486468

  5. Leaf Treatments with a Protein-Based Resistance Inducer Partially Modify Phyllosphere Microbial Communities of Grapevine.

    PubMed

    Cappelletti, Martina; Perazzolli, Michele; Antonielli, Livio; Nesler, Andrea; Torboli, Esmeralda; Bianchedi, Pier L; Pindo, Massimo; Puopolo, Gerardo; Pertot, Ilaria

    2016-01-01

    Protein derivatives and carbohydrates can stimulate plant growth, increase stress tolerance, and activate plant defense mechanisms. However, these molecules can also act as a nutritional substrate for microbial communities living on the plant phyllosphere and possibly affect their biocontrol activity against pathogens. We investigated the mechanisms of action of a protein derivative (nutrient broth, NB) against grapevine downy mildew, specifically focusing on the effects of foliar treatments on plant defense stimulation and on the composition and biocontrol features of the phyllosphere microbial populations. NB reduced downy mildew symptoms and induced the expression of defense-related genes in greenhouse- and in vitro-grown plants, indicating the activation of grapevine resistance mechanisms. Furthermore, NB increased the number of culturable phyllosphere bacteria and altered the composition of bacterial and fungal populations on leaves of greenhouse-grown plants. Although, NB-induced changes on microbial populations were affected by the structure of indigenous communities originally residing on grapevine leaves, degrees of disease reduction and defense gene modulation were consistent among the experiments. Thus, modifications in the structure of phyllosphere populations caused by NB application could partially contribute to downy mildew control by competition for space or other biocontrol strategies. Particularly, changes in the abundance of phyllosphere microorganisms may provide a contribution to resistance induction, partially affecting the hormone-mediated signaling pathways involved. Modifying phyllosphere populations by increasing natural biocontrol agents with the application of selected nutritional factors can open new opportunities in terms of sustainable plant protection strategies. PMID:27486468

  6. Effects of canola seed supplementation on intake, digestion, duodenal protein supply, and microbial efficiency in steers fed forage-based diets.

    PubMed

    Leupp, J L; Lardy, G P; Soto-Navarro, S A; Bauer, M L; Caton, J S

    2006-02-01

    Fourteen Holstein steers (446 +/- 4.4 kg of initial BW) with ruminal, duodenal, and ileal cannulas were used in a completely randomized design to evaluate effects of whole or ground canola seed (23.3% CP and 39.6% ether extract; DM basis) on intake, digestion, duodenal protein supply, and microbial efficiency in steers fed low-quality hay. Our hypothesis was that processing would be necessary to optimize canola use in diets based on low-quality forage. The basal diet consisted of ad libitum access to switchgrass hay (5.8% CP; DM basis) offered at 0700 daily. Treatments consisted of hay only (control), hay plus whole canola (8% of dietary DM), or hay plus ground canola (8% of dietary DM). Supplemental canola was provided based on the hay intake of the previous day. Steers were adapted to diets for 14 d followed by a 7-d collection period. Total DMI, OM intake, and OM digestibility were not affected (P > or = 0.31) by treatment. Similarly, no differences (P > or = 0.62) were observed for NDF or ADF total tract digestion. Bacterial OM at the duodenum increased (P = 0.01) with canola-containing diets compared with the control diet and increased (P = 0.08) in steers consuming ground canola compared with whole canola. Apparent and true ruminal CP digestibilities were increased (P = 0.01) with canola supplementation compared with the control diet. Canola supplementation decreased ruminal pH (P = 0.03) compared with the control diet. The molar proportion of acetate in the rumen tended (P = 0.10) to decrease with canola supplementation. The molar proportion of acetate in ruminal fluid decreased (P = 0.01), and the proportion of propionate increased (P = 0.01), with ground canola compared with whole canola. In situ disappearance rate of hay DM, NDF, and ADF were not altered by treatment (P > or = 0.32). In situ disappearance rate of canola DM, NDF, and ADF increased (P = 0.01) for ground canola compared with whole canola. Similarly, ground canola had greater (P = 0

  7. Insights into resistome and stress responses genes in Bubalus bubalis rumen through metagenomic analysis.

    PubMed

    Reddy, Bhaskar; Singh, Krishna M; Patel, Amrutlal K; Antony, Ancy; Panchasara, Harshad J; Joshi, Chaitanya G

    2014-10-01

    Buffalo rumen microbiota experience variety of diets and represents a huge reservoir of mobilome, resistome and stress responses. However, knowledge of metagenomic responses to such conditions is still rudimentary. We analyzed the metagenomes of buffalo rumen in the liquid and solid phase of the rumen biomaterial from river buffalo adapted to varying proportion of concentrate to green or dry roughages, using high-throughput sequencing to know the occurrence of antibiotics resistance genes, genetic exchange between bacterial population and environmental reservoirs. A total of 3914.94 MB data were generated from all three treatments group. The data were analysed with Metagenome rapid annotation system tools. At phyla level, Bacteroidetes were dominant in all the treatments followed by Firmicutes. Genes coding for functional responses to stress (oxidative stress and heat shock proteins) and resistome genes (resistance to antibiotics and toxic compounds, phages, transposable elements and pathogenicity islands) were prevalent in similar proportion in liquid and solid fraction of rumen metagenomes. The fluoroquinolone resistance, MDR efflux pumps and Methicillin resistance genes were broadly distributed across 11, 9, and 14 bacterial classes, respectively. Bacteria responsible for phages replication and prophages and phage packaging and rlt-like streptococcal phage genes were mostly assigned to phyla Bacteroides, Firmicutes and proteaobacteria. Also, more reads matching the sigma B genes were identified in the buffalo rumen. This study underscores the presence of diverse mechanisms of adaptation to different diet, antibiotics and other stresses in buffalo rumen, reflecting the proportional representation of major bacterial groups. PMID:24985977

  8. Diverse and divergent protein post-translational modifications in two growth stages of a natural microbial community

    SciTech Connect

    Li, Zhou; Wang, Yingfeng; Yao, Qiuming; Justice, Nicholas B.; Ahn, Tae-Hyuk; Xu, Dong; Hettich, Robert {Bob} L; Banfield, Jillian F.; Pan, Chongle

    2014-01-01

    Detailed characterization of posttranslational modifications (PTMs) of proteins in microbial communities remains a significant challenge. Here we directly identify and quantify a broad range of PTMs (hydroxylation, methylation, citrullination, acetylation, phosphorylation, methylthiolation, S-nitrosylation and nitration) in a natural microbial community from an acid mine drainage site. Approximately 29% of the identified proteins of the dominant Leptospirillum group II bacteria are modified, and 43% of modified proteins carry multiple PTM types. Most PTM events, except S-nitrosylations, have low fractional occupancy. Notably, PTM events are detected on Cas proteins involved in antiviral defense, an aspect of Cas biochemistry not considered previously. Further, Cas PTM profiles from Leptospirillum group II differ in early versus mature biofilms. PTM patterns are divergent on orthologues of two closely related, but ecologically differentiated, Leptospirillum group II bacteria. Our results highlight the prevalence and dynamics of PTMs of proteins, with potential significance for ecological adaptation and microbial evolution.

  9. Quantifying the Responses of Mixed Rumen Microbes to Excess Carbohydrate

    PubMed Central

    Hackmann, Timothy J.; Diese, Leanne E.

    2013-01-01

    The aim of this study was to determine if a mixed microbial community from the bovine rumen would respond to excess carbohydrate by accumulating reserve carbohydrate, energy spilling (dissipating excess ATP energy as heat), or both. Mixed microbes from the rumen were washed with N-free buffer and dosed with glucose. Total heat production was measured by calorimetry. Energy spilling was calculated as heat production not accounted by (i) endogenous metabolism (heat production before dosing glucose) and (ii) synthesis of reserve carbohydrate (heat from synthesis itself and reactions yielding ATP for it). For cells dosed with 5 mM glucose, synthesis of reserve carbohydrate and endogenous metabolism accounted for nearly all heat production (93.7%); no spilling was detected (P = 0.226). For cells dosed with 20 mM glucose, energy spilling was not detected immediately after dosing, but it became significant (P < 0.05) by approximately 30 min after dosing with glucose. Energy spilling accounted for as much as 38.7% of heat production in one incubation. Nearly all energy (97.9%) and carbon (99.9%) in glucose were recovered in reserve carbohydrate, fermentation acids, CO2, CH4, and heat. This full recovery indicates that products were measured completely and that spilling was not a methodological artifact. These results should aid future research aiming to mechanistically account for variation in energetic efficiency of mixed microbial communities. PMID:23584777

  10. High Potential Source for Biomass Degradation Enzyme Discovery and Environmental Aspects Revealed through Metagenomics of Indian Buffalo Rumen

    PubMed Central

    Singh, K. M.; Reddy, Bhaskar; Patel, Dishita; Patel, A. K.; Patel, J. B.; Joshi, C. G.

    2014-01-01

    The complex microbiomes of the rumen functions as an effective system for plant cell wall degradation, and biomass utilization provide genetic resource for degrading microbial enzymes that could be used in the production of biofuel. Therefore the buffalo rumen microbiota was surveyed using shot gun sequencing. This metagenomic sequencing generated 3.9 GB of sequences and data were assembled into 137270 contiguous sequences (contigs). We identified potential 2614 contigs encoding biomass degrading enzymes including glycoside hydrolases (GH: 1943 contigs), carbohydrate binding module (CBM: 23 contigs), glycosyl transferase (GT: 373 contigs), carbohydrate esterases (CE: 259 contigs), and polysaccharide lyases (PE: 16 contigs). The hierarchical clustering of buffalo metagenomes demonstrated the similarities and dissimilarity in microbial community structures and functional capacity. This demonstrates that buffalo rumen microbiome was considerably enriched in functional genes involved in polysaccharide degradation with great prospects to obtain new molecules that may be applied in the biofuel industry. PMID:25136572

  11. High potential source for biomass degradation enzyme discovery and environmental aspects revealed through metagenomics of Indian buffalo rumen.

    PubMed

    Singh, K M; Reddy, Bhaskar; Patel, Dishita; Patel, A K; Parmar, Nidhi; Patel, Anand; Patel, J B; Joshi, C G

    2014-01-01

    The complex microbiomes of the rumen functions as an effective system for plant cell wall degradation, and biomass utilization provide genetic resource for degrading microbial enzymes that could be used in the production of biofuel. Therefore the buffalo rumen microbiota was surveyed using shot gun sequencing. This metagenomic sequencing generated 3.9 GB of sequences and data were assembled into 137270 contiguous sequences (contigs). We identified potential 2614 contigs encoding biomass degrading enzymes including glycoside hydrolases (GH: 1943 contigs), carbohydrate binding module (CBM: 23 contigs), glycosyl transferase (GT: 373 contigs), carbohydrate esterases (CE: 259 contigs), and polysaccharide lyases (PE: 16 contigs). The hierarchical clustering of buffalo metagenomes demonstrated the similarities and dissimilarity in microbial community structures and functional capacity. This demonstrates that buffalo rumen microbiome was considerably enriched in functional genes involved in polysaccharide degradation with great prospects to obtain new molecules that may be applied in the biofuel industry. PMID:25136572

  12. High potential source for biomass degradation enzyme discovery and environmental aspects revealed through metagenomics of Indian buffalo rumen.

    PubMed

    Singh, K M; Reddy, Bhaskar; Patel, Dishita; Patel, A K; Parmar, Nidhi; Patel, Anand; Patel, J B; Joshi, C G

    2014-01-01

    The complex microbiomes of the rumen functions as an effective system for plant cell wall degradation, and biomass utilization provide genetic resource for degrading microbial enzymes that could be used in the production of biofuel. Therefore the buffalo rumen microbiota was surveyed using shot gun sequencing. This metagenomic sequencing generated 3.9 GB of sequences and data were assembled into 137270 contiguous sequences (contigs). We identified potential 2614 contigs encoding biomass degrading enzymes including glycoside hydrolases (GH: 1943 contigs), carbohydrate binding module (CBM: 23 contigs), glycosyl transferase (GT: 373 contigs), carbohydrate esterases (CE: 259 contigs), and polysaccharide lyases (PE: 16 contigs). The hierarchical clustering of buffalo metagenomes demonstrated the similarities and dissimilarity in microbial community structures and functional capacity. This demonstrates that buffalo rumen microbiome was considerably enriched in functional genes involved in polysaccharide degradation with great prospects to obtain new molecules that may be applied in the biofuel industry.

  13. Rumen conditions that predispose cattle to pasture bloat.

    PubMed

    Majak, W; Howarth, R E; Cheng, K J; Hall, J W

    1983-08-01

    Rumen contents from the dorsal sac were examined before alfalfa ingestion to determine factors that predispose cattle to pasture bloat. Chlorophyll concentration, buoyancy of particulate matter, and rates of gas production were significantly higher in cattle that subsequently bloated than in those that did not. Higher chlorophyll in bloat cases indicated accumulation of suspended chloroplast particles in the dorsal sac, perhaps due to increased buoyancy of the particulate matter. The higher fermentation rates (in the presence of glucose) suggested that the latent capacity for gas production was due to microbial colonization of suspended feed particles. Chlorophyll 4 h after feeding was also higher in bloated as compared to unbloated animals. In short, the microbial colonization and retention of particulate matter provided active inocula for promoting rapid legume digestion. Consequently, gas production was enhanced when feeding commenced, but the fermentation gases were trapped by the buoyant, frothy ingesta, resulting in the condition of pasture bloat. PMID:6619348

  14. Impact of subacute ruminal acidosis (SARA) adaptation on rumen microbiota in dairy cattle using pyrosequencing.

    PubMed

    Mao, S Y; Zhang, R Y; Wang, D S; Zhu, W Y

    2013-12-01

    The objective of this study was to evaluate the changes in bacterial populations in the rumen of dairy cattle following adaptation to subacute ruminal acidosis (SARA) using 16S rRNA gene pyrosequencing. Rumen contents were collected from four cattle adapted to either a 40% (control diet, COD) or 70% (SARA induction diet, SAID) concentrate feeds. DNA was extracted from each of the samples. Bacterial 16S rRNA genes of ruminal DNA extracts were PCR amplified with 2 bar coded primer sets and sequenced by 454 pyrosequencing. At a high taxonomic level, the percentage of Proteobacteria and Bacteroidetes were reduced by SAID feeding, whereas Firmicutes and Actinobacteria were more abundant in the SAID than in the COD group. At the genus level, as compared with the COD group, the abundances of Prevotella, Treponema, Anaeroplasma, Papillibacter, Acinetobacter and unclassified populations including unclassified Lentisphaerae, and unclassified bacteria were lower (P < 0.05), while the percentages of Ruminococcus, Atopobium, unclassified Clostridiales and Bifidobacterium were increased (P < 0.05) in the SAID group. Feeding of SAID reduced (P < 0.001) the diversity of the rumen microbial community. Taken together, our findings provide a comprehensive picture of current knowledge of the community structure of the rumen bacterial ecosystem during SARA, and enhance our understanding about the ruminal microbial ecology that may be useful in the prevention of ruminal acidosis.

  15. Metagenomic analysis of virulence-associated and antibiotic resistance genes of microbes in rumen of Indian buffalo (Bubalus bubalis).

    PubMed

    Singh, K M; Jakhesara, S J; Koringa, P G; Rank, D N; Joshi, C G

    2012-10-10

    A major research goal in rumen microbial ecology is to understand the relationship between community composition and its function, particularly involved in fermentation process is of a potential interest. The buffalo rumen microbiota impacts human food safety as well as animal health. Although the bacteria of bovine rumen have been well characterized, techniques have been lacking to correlate total community structure with gene function. We applied 454 next generations sequencing technology to characterize general microbial diversity present in buffalo rumen metagenome and also identified the repertoire of microbial genes present, including genes associated with antibiotic resistance and bacterial virulence. Results suggest that over six percent (6.44%) of the sequences from our buffalo rumen pool sample could be categorized as virulence genes and genes associated with resistance to antibiotic and toxic compounds (RATC), which is a higher proportion of virulence genes reported from metagenome samples of chicken cecum (5.39%), cow rumen (4.43%) and Sargasso sea (2.95%). However, it was lower than the proportion found in cow milk (11.33%) cattle faeces (8.4%), Antarctic marine derived lake (8.45%), human fecal (7.7%) and farm soil (7.79%). The dynamic nature of metagenomic data, together with the large number of RATC classes observed in samples from widely different ecologies indicates that metagenomic data can be used to track potential targets and relative amounts of antibiotic resistance genes in individual animals. In addition, these data can be also used to generate antibiotic resistance gene profiles to facilitate an understanding of the ecology of the microbial communities in each habitat as well as the epidemiology of antibiotic resistant gene transport between and among habitats.

  16. Metagenomic analysis of virulence-associated and antibiotic resistance genes of microbes in rumen of Indian buffalo (Bubalus bubalis).

    PubMed

    Singh, K M; Jakhesara, S J; Koringa, P G; Rank, D N; Joshi, C G

    2012-10-10

    A major research goal in rumen microbial ecology is to understand the relationship between community composition and its function, particularly involved in fermentation process is of a potential interest. The buffalo rumen microbiota impacts human food safety as well as animal health. Although the bacteria of bovine rumen have been well characterized, techniques have been lacking to correlate total community structure with gene function. We applied 454 next generations sequencing technology to characterize general microbial diversity present in buffalo rumen metagenome and also identified the repertoire of microbial genes present, including genes associated with antibiotic resistance and bacterial virulence. Results suggest that over six percent (6.44%) of the sequences from our buffalo rumen pool sample could be categorized as virulence genes and genes associated with resistance to antibiotic and toxic compounds (RATC), which is a higher proportion of virulence genes reported from metagenome samples of chicken cecum (5.39%), cow rumen (4.43%) and Sargasso sea (2.95%). However, it was lower than the proportion found in cow milk (11.33%) cattle faeces (8.4%), Antarctic marine derived lake (8.45%), human fecal (7.7%) and farm soil (7.79%). The dynamic nature of metagenomic data, together with the large number of RATC classes observed in samples from widely different ecologies indicates that metagenomic data can be used to track potential targets and relative amounts of antibiotic resistance genes in individual animals. In addition, these data can be also used to generate antibiotic resistance gene profiles to facilitate an understanding of the ecology of the microbial communities in each habitat as well as the epidemiology of antibiotic resistant gene transport between and among habitats. PMID:22850272

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

    PubMed

    Chen, X B; Hovell, F D; Orskov, E R

    1990-03-01

    The saliva of sheep was shown to contain significant concentrations of uric acid (16(SD 4.5) mumol/l) and allantoin (120(SD 16.4) mumol/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 allantion 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.

  18. The Involvement of Microbially Derived Extracellular Proteins in Nanoparticle Formation and Aggregation

    NASA Astrophysics Data System (ADS)

    Pearce, C. I.; Moreau, J. W.

    2007-12-01

    spheroids. Analysis of these spheroids revealed a formative association between ZnS nanoparticles and microbially derived extracellular proteins. Direct protein extraction and isolation yielded a dominant ~37 kDa band for ongoing mass spectrometry characterization. Parallel experimental simulations of nanoparticle-amino acid interactions suggest an important role for cysteine-bearing proteins in promoting the aggregation of nanoparticulate metal-sulfides. Identification of microbial proteins that interact with natural or synthetic nanoparticles provides potential for synthesizing specific peptide sequences for technological and environmental applications, and sheds light into environmental nanoscale biomineralization processes that may significantly impact water quality.

  19. Effects of replacing canola meal as the major protein source with wheat dried distillers grains with solubles on ruminal function, microbial protein synthesis, omasal flow, and milk production in cows.

    PubMed

    Chibisa, G E; Christensen, D A; Mutsvangwa, T

    2012-02-01

    bacterial nonammonia N (NAN) and bacterial efficiency (g of total bacterial NAN flow/kg of organic matter truly digested in the rumen) were not different among diets; however, feeding W-DDGS resulted in a quadratic increase in nonammonia nonbacterial N flow at the omasal canal (271, 318, 336, and 311 g/d for 0, 10, 15, and 20% W-DDGS, respectively). These data indicate that W-DDGS can substitute for CM as the major protein source in dairy cow diets without negatively affecting ruminal fermentation, microbial protein production, and omasal nutrient flow, and can potentially increase dry matter intake and milk yield.

  20. RNA-Seq detection of differential gene expression in the rumen of beef steers associated with feed efficiency phenotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The efficient utilization of feedstuffs is an economically important trait in beef production. The rumen is important to the digestive process of steers interacting with feed, microbial populations, and volatile fatty acids indicating it may play a critical role in feed efficiency. To gain an unders...

  1. Microbial Physiology of the Conversion of Residual Oil to Methane: A Protein Prospective

    NASA Astrophysics Data System (ADS)

    Morris, Brandon E. L.; Bastida-Lopez, Felipe; von Bergen, Martin; Richnow, Hans-Hermann; Suflita, Joseph M.

    2010-05-01

    Traditional petroleum recovery techniques are unable to extract the majority of oil in most petroliferous deposits. The recovery of even a fraction of residual hydrocarbon in conventional reserves could represent a substantive energy supply. To this end, the microbial conversion of residual oil to methane has gained increasing relevance in recent years [1,2]. Worldwide demand for methane is expected to increase through 2030 [3], as it is a cleaner-burning alternative to traditional fuels [4]. To investigate the microbial physiology of hydrocarbon-decomposition and ultimate methanogenesis, we initiated a two-pronged approach. First, a model alkane-degrading sulfate-reducing bacterium, Desulfoglaeba alkanexedens, was used to interrogate the predominant metabolic pathway(s) differentially expressed during growth on either n-decane or butyrate. A total of 81 proteins were differentially expressed during bacterial growth on butyrate, while 100 proteins were unique to the alkane-grown condition. Proteins related to alkylsuccinate synthase, or the homologous 1-methyl alkylsuccinate synthase, were identified only in the presence of the hydrocarbon. Secondly, we used a newly developed stable isotope probing technique [5] targeted towards proteins to monitor the flux of carbon through a residual oil-degrading bacterial consortium enriched from a gas-condensate contaminated aquifer [1]. Combined carbon and hydrogen stable isotope fractionation identified acetoclastic methanogenesis as the dominant process in this system. Such findings agree with the previous clone library characterization of the consortium. Furthermore, hydrocarbon activation was determined to be the rate-limiting process during the net conversion of residual oil to methane. References 1. Gieg, L.M., K.E. Duncan, and J.M. Suflita, Bioenegy production via microbial conversion of residual oil to natural gas. Appl Environ Micro, 2008. 74(10): p. 3022-3029. 2. Jones, D.M., et al., Crude-oil biodegradation via

  2. Proteomic profiling of microbial transglutaminase-induced polymerization of milk proteins.

    PubMed

    Hsieh, J F; Pan, P H

    2012-02-01

    Microbial transglutaminase (MTGase)-induced polymerization of individual milk proteins during incubation was investigated using a proteomics-based approach. The addition of MTGase (0.25-2.0 units/mL) caused the milk proteins to polymerize after a 3-h incubation period. Sodium dodecyl sulfate-PAGE analysis showed that the total intensities of the protein bands that corresponded to α(S)-casein, β-casein, and κ-casein decreased from 8,245.6, 6,677.2, and 586.6 arbitrary units to 1,911.7, 0.0, and 66.2 arbitrary units, respectively. Components with higher molecular weights were observed, and the intensity of these proteins increased after 3h of incubation. These results support that inter- or intramolecular crosslinking occurred in the casein proteins of MTGase-treated milk. Two-dimensional electrophoresis analysis indicated that isomers of β-casein, κ-casein, a fraction of serum albumin, α(S1)-casein, α(S2)-casein, β-lactoglobulin, and α-lactalbumin in the milk were polymerized following incubation with MTGase. In addition, MTGase-induced polymerization occurred earlier for β-casein and κ-casein isomers than for other milk proteins.

  3. Influence of dietary docosahexaenoic acid supplementation on the overall rumen microbiota of dairy cows and linkages with production parameters.

    PubMed

    Torok, Valeria A; Percy, Nigel J; Moate, Peter J; Ophel-Keller, Kathy

    2014-05-01

    The rumen microbiota contributes to greenhouse gas emissions and has an impact on feed efficiency and ruminant product fatty acid composition. Dietary fat supplements have shown promise in reducing enteric methane production and in altering the fatty acid profiles of ruminant-derived products, yet in vivo studies on how these impact the rumen microbiota are limited. In this study, we investigated the rumen bacterial, archaeal, fungal, and ciliate protozoan communities of dairy cows fed diets supplemented with 4 levels of docosahexaenoic acid (DHA) (0, 25, 50, and 75 g·cow(-1)·day(-1)) and established linkages between microbial communities and production parameters. Supplementation with DHA significantly (P < 0.05) altered rumen bacterial and archaeal, including methanogenic archaeal, communities but had no significant (P > 0.05) effects on rumen fungal or ciliate protozoan communities. Rumen bacterial communities of cows receiving no DHA were correlated with increased saturated fatty acids (C18:0 and C11:0) in their milk. Furthermore, rumen bacterial communities of cows receiving a diet supplemented with 50 g DHA·cow(-1)·day(-1) were correlated with increases in monounsaturated fatty acids (C20:1n-9) and polyunsaturated fatty acids (C22:5n-3; C22:6n-3; C18:2 cis-9, trans-11; C22:3n-6; and C18:2n-6 trans) in their milk. The significant diet-associated changes in rumen archaeal communities observed did not result in altered enteric methane outputs in these cows.

  4. Effects of feeding Mediterranean buffalo sorghum silage versus maize silage on the rumen microbiota and milk fatty acid content.

    PubMed

    Ann Huws, Sharon; Chiariotti, Antonella; Sarubbi, Fiorella; Carfì, Francesca; Pace, Vilma

    2012-01-01

    Sorghum presents a sustainable feedstock for Mediterranean buffaloes due to its reduced water and nitrogen requirements compared with maize, which is currently fed primarily. We investigated the effects of feeding sorghum as opposed to maize on Mediterranean buffalo rumen microbial diversity and milk fatty acid content. Four cannulated lactating Mediterranean buffalo cows were fed a basal diet for one month before switching either to maize or sorghum-silage based diets for a 3-month period. Buffaloes were then changed over to the contrasting diet for a further one month. Rumen and milk samples were collected at the end of each month. DGGE- and T-RFLP-based dendrograms generated from rumen samples did not show an effect of diet on rumen bacterial diversity. Milk samples also did not differ in terms of their fatty acid content post sorghum feeding as compared with maize feeding. Thus, sorghum provides an environmentally beneficial alternative to maize for feeding Mediterranean buffalo with little effect on rumen microbial diversity or milk fatty acid composition compared with maize feeding.

  5. Effects of feeding Mediterranean buffalo sorghum silage versus maize silage on the rumen microbiota and milk fatty acid content.

    PubMed

    Ann Huws, Sharon; Chiariotti, Antonella; Sarubbi, Fiorella; Carfì, Francesca; Pace, Vilma

    2012-01-01

    Sorghum presents a sustainable feedstock for Mediterranean buffaloes due to its reduced water and nitrogen requirements compared with maize, which is currently fed primarily. We investigated the effects of feeding sorghum as opposed to maize on Mediterranean buffalo rumen microbial diversity and milk fatty acid content. Four cannulated lactating Mediterranean buffalo cows were fed a basal diet for one month before switching either to maize or sorghum-silage based diets for a 3-month period. Buffaloes were then changed over to the contrasting diet for a further one month. Rumen and milk samples were collected at the end of each month. DGGE- and T-RFLP-based dendrograms generated from rumen samples did not show an effect of diet on rumen bacterial diversity. Milk samples also did not differ in terms of their fatty acid content post sorghum feeding as compared with maize feeding. Thus, sorghum provides an environmentally beneficial alternative to maize for feeding Mediterranean buffalo with little effect on rumen microbial diversity or milk fatty acid composition compared with maize feeding. PMID:22688241

  6. Peptidoglycan-associated outer membrane protein Mep45 of rumen anaerobe Selenomonas ruminantium forms a non-specific diffusion pore via its C-terminal transmembrane domain.

    PubMed

    Kojima, Seiji; Hayashi, Kanako; Tochigi, Saeko; Kusano, Tomonobu; Kaneko, Jun; Kamio, Yoshiyuki

    2016-10-01

    The major outer membrane protein Mep45 of Selenomonas ruminantium, an anaerobic Gram-negative bacterium, comprises two distinct domains: the N-terminal S-layer homologous (SLH) domain that protrudes into the periplasm and binds to peptidoglycan, and the remaining C-terminal transmembrane domain, whose function has been unknown. Here, we solubilized and purified Mep45 and characterized its function using proteoliposomes reconstituted with Mep45. We found that Mep45 forms a nonspecific diffusion channel via its C-terminal region. The channel was permeable to solutes smaller than a molecular weight of roughly 600, and the estimated pore radius was 0.58 nm. Truncation of the SLH domain did not affect the channel property. On the basis of the fact that Mep45 is the most abundant outer membrane protein in S. ruminantium, we conclude that Mep45 serves as a main pathway through which small solutes diffuse across the outer membrane of this bacterium.

  7. Expanded microbial genome coverage and improved protein family annotation in the COG database

    PubMed Central

    Galperin, Michael Y.; Makarova, Kira S.; Wolf, Yuri I.; Koonin, Eugene V.

    2015-01-01

    Microbial genome sequencing projects produce numerous sequences of deduced proteins, only a small fraction of which have been or will ever be studied experimentally. This leaves sequence analysis as the only feasible way to annotate these proteins and assign to them tentative functions. The Clusters of Orthologous Groups of proteins (COGs) database (http://www.ncbi.nlm.nih.gov/COG/), first created in 1997, has been a popular tool for functional annotation. Its success was largely based on (i) its reliance on complete microbial genomes, which allowed reliable assignment of orthologs and paralogs for most genes; (ii) orthology-based approach, which used the function(s) of the characterized member(s) of the protein family (COG) to assign function(s) to the entire set of carefully identified orthologs and describe the range of potential functions when there were more than one; and (iii) careful manual curation of the annotation of the COGs, aimed at detailed prediction of the biological function(s) for each COG while avoiding annotation errors and overprediction. Here we present an update of the COGs, the first since 2003, and a comprehensive revision of the COG annotations and expansion of the genome coverage to include representative complete genomes from all bacterial and archaeal lineages down to the genus level. This re-analysis of the COGs shows that the original COG assignments had an error rate below 0.5% and allows an assessment of the progress in functional genomics in the past 12 years. During this time, functions of many previously uncharacterized COGs have been elucidated and tentative functional assignments of many COGs have been validated, either by targeted experiments or through the use of high-throughput methods. A particularly important development is the assignment of functions to several widespread, conserved proteins many of which turned out to participate in translation, in particular rRNA maturation and tRNA modification. The new version of the

  8. [The effect of cadmium on the protozoan population and rumen fermentation of feed in an artificial rumen].

    PubMed

    Jalc, D; Kisidayová, S; Siroka, P; Sviatko, P

    1994-01-01

    In our experiment, the effects of 5, 10 and 20 mg cadmium per kg dry matter (DM) on protozoan population and rumen fermentation of feed ration consisting of 11.7 g DM of hay and 2.8 g DM of barley (80: 20%) were followed in artificial rumen (Rusitec). The results of the experiment showed that the addition of 5 and 10 mg cadmium per kg DM less significantly (P < 0.05) decreased the digestibility of DM, organic matter and neutral detergent fibre and significantly (P < 0.01) decreased the digestibility of cellulose of feed ration. It is interesting that the addition of 20 mg cadmium per kg DM did not influence these parameters. Although the total gas production showed a tendency of decrease due to the influence of the addition of 5, 10 and 20 mg cadmium/kg DM, methane and CO2 production was not altered. However, cadmium significantly influenced the production of individual volatile fatty acids-VFA's (mmol/day). The addition of 5, 10 and 20 mg cadmium per kg DM significantly increased acetic acid production (by 10-20%) and decreased propionic acid production (by 18-30%). Therefore, the acetate: propionate ratio was significantly increased (from 1.57 to 2.45-2.60) by the addition of cadmium. The production of n-valeric and iso-valeric acids was also significantly decreased in comparison with the control. The decrease of individual VFA's was more significant at the higher amount of cadmium added into the fermentation system. The proportions of individual VFA's in total VFA production expressed in molar % showed a similar character. The changes in the production of individual VFA's due to the influence of the addition of 5, 10 and 20 mg cadmium per kg DM caused a decrease in energetic efficiency of VFA's (P < 0.001) in comparison with the control. The other parameters of rumen fermentation - utilization of glucose, adenosine triphosphate (ATP) production, fermented hexose, fermented amino acids and fermented organic matter (OMF) were not influenced by the cadmium addition

  9. PATtyFams: Protein families for the microbial genomes in the PATRIC database

    DOE PAGESBeta

    Davis, James J.; Gerdes, Svetlana; Olsen, Gary J.; Olson, Robert; Pusch, Gordon D.; Shukla, Maulik; Vonstein, Veronika; Wattam, Alice R.; Yoo, Hyunseung

    2016-02-08

    The ability to build accurate protein families is a fundamental operation in bioinformatics that influences comparative analyses, genome annotation, and metabolic modeling. For several years we have been maintaining protein families for all microbial genomes in the PATRIC database (Pathosystems Resource Integration Center, patricbrc.org) in order to drive many of the comparative analysis tools that are available through the PATRIC website. However, due to the burgeoning number of genomes, traditional approaches for generating protein families are becoming prohibitive. In this report, we describe a new approach for generating protein families, which we call PATtyFams. This method uses the k-mer-based functionmore » assignments available through RAST (Rapid Annotation using Subsystem Technology) to rapidly guide family formation, and then differentiates the function-based groups into families using a Markov Cluster algorithm (MCL). In conclusion, this new approach for generating protein families is rapid, scalable and has properties that are consistent with alignment-based methods.« less

  10. Peptidoglycan-associated outer membrane protein Mep45 of rumen anaerobe Selenomonas ruminantium forms a non-specific diffusion pore via its C-terminal transmembrane domain

    PubMed Central

    Kojima, Seiji; Hayashi, Kanako; Tochigi, Saeko; Kusano, Tomonobu; Kaneko, Jun; Kamio, Yoshiyuki

    2016-01-01

    The major outer membrane protein Mep45 of Selenomonas ruminantium, an anaerobic Gram-negative bacterium, comprises two distinct domains: the N-terminal S-layer homologous (SLH) domain that protrudes into the periplasm and binds to peptidoglycan, and the remaining C-terminal transmembrane domain, whose function has been unknown. Here, we solubilized and purified Mep45 and characterized its function using proteoliposomes reconstituted with Mep45. We found that Mep45 forms a nonspecific diffusion channel via its C-terminal region. The channel was permeable to solutes smaller than a molecular weight of roughly 600, and the estimated pore radius was 0.58 nm. Truncation of the SLH domain did not affect the channel property. On the basis of the fact that Mep45 is the most abundant outer membrane protein in S. ruminantium, we conclude that Mep45 serves as a main pathway through which small solutes diffuse across the outer membrane of this bacterium. PMID:27310312

  11. Passive mechanical properties of ovine rumen tissue

    NASA Astrophysics Data System (ADS)

    Waite, Stephen J.; Cater, John E.; Walker, Cameron G.; Amirapu, Satya; Waghorn, Garry C.; Suresh, Vinod

    2016-05-01

    Mechanical and structural properties of ovine rumen tissue have been determined using uniaxial tensile testing of tissue from four animals at five rumen locations and two orientations. Animal and orientation did not have a significant effect on the stress-strain response, but there was a significant difference between rumen locations. Histological studies showed two orthogonal muscle layers in all regions except the reticulum, which has a more isotropic structure. A quasi-linear viscoelastic model was fitted to the relaxation stage for each region. Model predictions of the ramp stage had RMS errors of 13-24% and were within the range of the experimental data.

  12. Exploring the Goat Rumen Microbiome from Seven Days to Two Years

    PubMed Central

    Wang, Lizhi; Xu, Qin; Kong, Fanli; Yang, Yindong; Wu, De; Mishra, Sudhanshu; Li, Ying

    2016-01-01

    Rumen microbial communities play important roles in feed conversion and the physiological development of the ruminants. Despite its significance, little is known about the rumen microbial communities at different life stages after birth. In this study, we characterized the rumen bacterial and the archaeal communities in 11 different age groups (7, 15, 30, 60, 90, 120, 150, 180, 360, 540 and 720 days old) of a crossbred F1 goats (n = 5 for each group) by using an Illumina MiSeq platform targeting the V3-V4 region of the 16S rRNA gene. We found that the bacterial communities were mainly composed of Bacteroidetes, Firmicutes, and Proteobacteria across all age groups. The relative abundance of Firmicutes was stable across all age groups. While changes in relative abundance were observed in Bacteroidetes and Proteobacteria, these two phyla reached a stable stage after weaning (day 90). Euryarchaeota (82%) and Thaumarchaeota (15%) were the dominant phyla of Archaea. Crenarchaeota was also observed, although at a very low relative abundance (0.68% at most). A clear age-related pattern was observed in the diversity of bacterial community with 59 OTUs associated with age. In contrast, no age-related OTU was observed in archaea. In conclusion, our results suggested that from 7 days to 2 years, the ruminal microbial community of our experimental goats underwent significant changes in response to the shift in age and diet. PMID:27135948

  13. Exploring the Goat Rumen Microbiome from Seven Days to Two Years.

    PubMed

    Wang, Lizhi; Xu, Qin; Kong, Fanli; Yang, Yindong; Wu, De; Mishra, Sudhanshu; Li, Ying

    2016-01-01

    Rumen microbial communities play important roles in feed conversion and the physiological development of the ruminants. Despite its significance, little is known about the rumen microbial communities at different life stages after birth. In this study, we characterized the rumen bacterial and the archaeal communities in 11 different age groups (7, 15, 30, 60, 90, 120, 150, 180, 360, 540 and 720 days old) of a crossbred F1 goats (n = 5 for each group) by using an Illumina MiSeq platform targeting the V3-V4 region of the 16S rRNA gene. We found that the bacterial communities were mainly composed of Bacteroidetes, Firmicutes, and Proteobacteria across all age groups. The relative abundance of Firmicutes was stable across all age groups. While changes in relative abundance were observed in Bacteroidetes and Proteobacteria, these two phyla reached a stable stage after weaning (day 90). Euryarchaeota (82%) and Thaumarchaeota (15%) were the dominant phyla of Archaea. Crenarchaeota was also observed, although at a very low relative abundance (0.68% at most). A clear age-related pattern was observed in the diversity of bacterial community with 59 OTUs associated with age. In contrast, no age-related OTU was observed in archaea. In conclusion, our results suggested that from 7 days to 2 years, the ruminal microbial community of our experimental goats underwent significant changes in response to the shift in age and diet. PMID:27135948

  14. Microbial transglutaminase-mediated synthesis of hapten-protein conjugates for immunoassays.

    PubMed

    Josten, A; Meusel, M; Spener, F

    1998-05-01

    Hapten-protein conjugates are essential in many immunochemical assays, in particular, in assays employing titration or competitive assay formats. By exploitation of the catalytic properties of the microbial transglutaminase from Streptoverticillium mobarense sp. (MTGase), i.e., acyl transfer between gamma-carboxamide groups and various primary amines, new techniques for the synthesis of hapten-protein conjugates were developed. This is demonstrated by two examples. The feasibility of MTGase for hapten-protein conjugate synthesis was studied by coupling the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) to casein. Different procedures for the synthesis and the immobilization of these 2,4-D-casein conjugates were evaluated, comprising (i) a batch procedure, (ii) coupling of 2,4-D to an already immobilized layer of casein, and (iii) a method for simultaneous immobilization and conjugation. Kinetic studies revealed that conjugate formation in the batch procedure was almost complete after approx 2 h. By employing the conjugates in a competitive ELISA, detection limits as low as 0.05 microgram/L 2,4-D were reached. Using the approach with simultaneous immobilization and conjugation, the time for the whole assay could be reduced to only 2 h. Finally, to demonstrate the versatility of the enzymatic synthesis of hapten-protein conjugates, an ELISA for 2,4,6-trinitrotoluene (TNT) determination based on transglutaminase-synthesized conjugates was developed. In this assay, a detection limit as low as 0.04 microgram/l TNT was obtained.

  15. Microbial metaproteomics: Identifying the repertoire of proteins that microorganisms use to compete and cooperate in complex environmental communities

    SciTech Connect

    Hettich, Robert {Bob} L; Sharma, Ritin; Chourey, Karuna; Giannone, Richard J

    2012-01-01

    The availability of genome information for microbial consortia, including unculturable species, from environmental samples has enabled systems-biology interrogation by providing a means to access genomic, transcriptomic, and proteomic information. This provides a unique opportunity to characterize the molecular activities and interactions of these microbial systems at a comprehensive level never before possible. Such information not only provides details about the organizational, functional, and metabolic activities of such systems, but also the untapped reserve of molecular activities that might be invoked and exploited under certain environmental conditions. Since bacteria naturally exist in complex ecosystems, it is imperative to develop and utilize analytical approaches that can provide molecular level details on systems consisting of mixed microbial membership. This is the realm of metaproteomics - the characterization of the complement of proteins expressed by a microbial community in an environmental sample

  16. Short-Term Protein Stable Isotope Probing of Microbial Communities to Associate Functions with Taxa (Invited)

    NASA Astrophysics Data System (ADS)

    Lipton, M. S.; Slysz, G. W.; Steinke, L. A.; Ward, D. M.; Klatt, C. G.; Clauss, T. R.; Purvine, S. O.; Anderson, G. A.; Payne, S. H.; Bryant, D. A.

    2013-12-01

    Determining which taxa in a community perform which functions is essential for understanding metabolite fluxes and metabolic interactions among community members. Specific taxa will alter their metabolism in order to acclimate to changing environmental factors such as light through the diel cycle, changing temperature and other factors. Monitoring which proteins are being expressed, and the quantitative protein expression patterns in the individual taxa as a response to external stimuli is key to understanding these mechanisms. Protein stable isotope probing (Pro-SIP) has strong potential for revealing key metabolizing taxa in complex microbial communities. In Pro-SIP studies, label incorporation is determined by the extent of the change in the isotopic profile of peptides when measured by mass spectrometry. While most Pro-SIP work to date has been performed under controlled laboratory conditions to allow extensive isotope labeling of the target organism(s), these techniques have not been applied to short term in situ studies due to the small degree of partial labeling of the proteins. We have applied Pro-SIP to study the assimilation of a labeled substrate into proteins to determine which taxa are responsible for sequestration of dissolved inorganic carbon in microbial mats associated with the alkaline siliceous hot springs of Yellowstone National Park. This community is fueled by sunlight as it transitions from dark to light; the aim was to understand the light-dependent pathway of inorganic carbon incorporation into different taxa during the early morning hours when the mat was in low light and anoxic. Each mat sample was incubated with 13C-bicarbonate for 3 h. Substrate assimilation was determined through standard proteomic techniques along with the use of SIPPER, a collection of algorithms that sensitively measure small changes in peptide isotopic patterns, allowing the determination of which taxa assimilated the substrate during this period. For the

  17. Effects of dietary supplementation of rumen-protected folic acid on rumen fermentation, degradability and excretion of urinary purine derivatives in growing steers.

    PubMed

    Wang, Cong; Liu, Qiang; Guo, Gang; Huo, WenJie; Ma, Le; Zhang, YanLi; Pei, CaiXia; Zhang, ShuanLin; Wang, Hao

    2016-12-01

    The present experiment was undertaken to determine the effects of dietary addition of rumen-protected folic acid (RPFA) on ruminal fermentation, nutrient degradability, enzyme activity and the relative quantity of ruminal cellulolytic bacteria in growing beef steers. Eight rumen-cannulated Jinnan beef steers averaging 2.5 years of age and 419 ± 1.9 kg body weight were used in a replicated 4 × 4 Latin square design. The four treatments comprised supplementation levels of 0 (Control), 70, 140 and 210 mg RPFA/kg dietary dry matter (DM). On DM basis, the ration consisted of 50% corn silage, 47% concentrate and 3% soybean oil. The DM intake (averaged 8.5 kg/d) was restricted to 95% of ad libitum intake. The intake of DM, crude protein (CP) and net energy for growth was not affected by treatments. In contrast, increasing RPFA supplementation increased average daily gain and the concentration of total volatile fatty acid and reduced ruminal pH linearly. Furthermore, increasing RPFA supplementation enhanced the acetate to propionate ratio and reduced the ruminal ammonia N content linearly. The ruminal effective degradability of neutral detergent fibre from corn silage and CP from concentrate improved linearly and was highest for the highest supplementation levels. The activities of cellobiase, xylanase, pectinase and α-amylase linearly increased, but carboxymethyl-cellulase and protease were not affected by the addition of RPFA. The relative quantities of Butyrivibrio fibrisolvens, Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes increased linearly. With increasing RPFA supplementation levels, the excretion of urinary purine derivatives was also increased linearly. The present results indicated that the supplementation of RPFA improved ruminal fermentation, nutrient degradability, activities of microbial enzymes and the relative quantity of the ruminal cellulolytic bacteria in a dose-dependent manner. According to the conditions of this

  18. PCR detection of uncultured rumen bacteria.

    PubMed

    Rosero, Jaime A; Strosová, Lenka; Mrázek, Jakub; Fliegerová, Kateřina; Kopečný, Jan

    2012-07-01

    16S rRNA sequences of ruminal uncultured bacterial clones from public databases were phylogenetically examined. The sequences were found to form two unique clusters not affiliated with any known bacterial species: cluster of unidentified sequences of free floating rumen fluid uncultured bacteria (FUB) and cluster of unidentified sequences of bacteria associated with rumen epithelium (AUB). A set of PCR primers targeting 16S rRNA of ruminal free uncultured bacteria and rumen epithelium adhering uncultured bacteria was designed based on these sequences. FUB primers were used for relative quantification of uncultured bacteria in ovine rumen samples. The effort to increase the population size of FUB group has been successful in sulfate reducing broth and culture media supplied with cellulose.

  19. Convergent Evolution of Rumen Microbiomes in High-Altitude Mammals.

    PubMed

    Zhang, Zhigang; Xu, Dongming; Wang, Li; Hao, Junjun; Wang, Jinfeng; Zhou, Xin; Wang, Weiwei; Qiu, Qiang; Huang, Xiaodan; Zhou, Jianwei; Long, Ruijun; Zhao, Fangqing; Shi, Peng

    2016-07-25

    Studies of genetic adaptation, a central focus of evolutionary biology, most often focus on the host's genome and only rarely on its co-evolved microbiome. The Qinghai-Tibetan Plateau (QTP) offers one of the most extreme environments for the survival of human and other mammalian species. Yaks (Bos grunniens) and Tibetan sheep (T-sheep) (Ovis aries) have adaptations for living in this harsh high-altitude environment, where nomadic Tibetan people keep them primarily for food and livelihood [1]. Adaptive evolution affects energy-metabolism-related genes in a way that helps these ruminants live at high altitude [2, 3]. Herein, we report convergent evolution of rumen microbiomes for energy harvesting persistence in two typical high-altitude ruminants, yaks and T-sheep. Both ruminants yield significantly lower levels of methane and higher yields of volatile fatty acids (VFAs) than their low-altitude relatives, cattle (Bos taurus) and ordinary sheep (Ovis aries). Ultra-deep metagenomic sequencing reveals significant enrichment in VFA-yielding pathways of rumen microbial genes in high-altitude ruminants, whereas methanogenesis pathways show enrichment in the cattle metagenome. Analyses of RNA transcriptomes reveal significant upregulation in 36 genes associated with VFA transport and absorption in the ruminal epithelium of high-altitude ruminants. Our study provides novel insights into the contributions of microbiomes to adaptive evolution in mammals and sheds light on the biological control of greenhouse gas emissions from livestock enteric fermentation. PMID:27321997

  20. Growth and survival of rumen fungi.

    PubMed

    Trinci, A P; Lowe, S E; Milne, A; Theodorou, M K

    1988-01-01

    The life cycle and growth kinetics of an anaerobic rumen fungus (Neocallimastix R1) in liquid and solid media are described, together with its response to light, temperature and oxygen. These results are discussed in relation to the survival of rumen fungi in saliva and faeces of sheep, and the possible routes for the transfer of anaerobic fungi between ruminants. The thallus and life cycle of Neocallimastix R1 are compared with those of aerobic chytrids.

  1. Studies of rumen function in an in vitro continuous culture system.

    PubMed

    Merry, R J; Smith, R H; McAllan, A B

    1987-06-01

    An in vitro continuous culture system to simulate processes in the rumen is described. This comprises a culture vessel (Fig. 2) fed continuously with pelleted solid feed (Fig. 1); artificial saliva also enters the vessel while effluent leaves it continuously in two streams, one is filtered and the other simply overflows (Fig. 1). In this way liquid and solid turnover times may be manipulated independently; in a trial experiment the former and latter were 6.3% and 3.4% per hour respectively. In four replicated experiments a steady-state was achieved in 5-6 days with a feed of barley, tapioca, fishmeal, urea and straw and maintained for a further 7 to 14 days. During a steady-state period, rumen characteristics in terms of protozoal count, pH, redox potential, total volatile fatty acid concentration and ammonia concentration were stable and similar to those found in the rumens of animals given similar diets. Flows of microbial N compounds were assessed by infused Na2H32PO4 with the artificial saliva and estimating the 32P label incorporated in bacterial fractions. The system demonstrably provided a satisfactory means for studying many aspects of rumen function.

  2. Cell Death Inducing Microbial Protein Phosphatase Inhibitors--Mechanisms of Action.

    PubMed

    Kleppe, Rune; Herfindal, Lars; Døskeland, Stein Ove

    2015-10-01

    Okadaic acid (OA) and microcystin (MC) as well as several other microbial toxins like nodularin and calyculinA are known as tumor promoters as well as inducers of apoptotic cell death. Their intracellular targets are the major serine/threonine protein phosphatases. This review summarizes mechanisms believed to be responsible for the death induction and tumor promotion with focus on the interdependent production of reactive oxygen species (ROS) and activation of Ca(2+)/calmodulin kinase II (CaM-KII). New data are presented using inhibitors of specific ROS producing enzymes to curb nodularin/MC-induced liver cell (hepatocyte) death. They indicate that enzymes of the arachidonic acid pathway, notably phospholipase A2, 5-lipoxygenase, and cyclooxygenases, may be required for nodularin/MC-induced (and presumably OA-induced) cell death, suggesting new ways to overcome at least some aspects of OA and MC toxicity. PMID:26506362

  3. Advancing understanding of microbial bioenergy conversion processes by activity-based protein profiling

    SciTech Connect

    Liu, Yun; Fredrickson, James K.; Sadler, Natalie C.; Nandhikonda, Premchendar; Smith, Richard D.; Wright, Aaron T.

    2015-09-25

    Here, the development of renewable biofuels is a global priority, but success will require novel technologies that greatly improve our understanding of microbial systems biology. An approach with great promise in enabling functional characterization of microbes is activity-based protein profiling (ABPP), which employs chemical probes to directly measure enzyme function in discrete enzyme classes in vivo and/or in vitro, thereby facilitating the rapid discovery of new biocatalysts and enabling much improved biofuel production platforms. We review general design strategies in ABPP, and highlight recent advances that are or could be pivotal to biofuels processes including applications of ABPP to cellulosic bioethanol, biodiesel, and phototrophic production of hydrocarbons. We also examine the key challenges and opportunities of ABPP in renewable biofuels research. The integration of ABPP with molecular and systems biology approaches will shed new insight on the catalytic and regulatory mechanisms of functional enzymes and their synergistic effects in the field of biofuels production.

  4. Production of Microbial Biomass Protein from Potato Processing Wastes by Cephalosporium eichhorniae

    PubMed Central

    Stevens, Coleen A.; Gregory, Kenneth F.

    1987-01-01

    The use of Cephalosporium eichhorniae 152 (ATCC 38255) (reclassified as Acremonium alabamense; see Addendum in Proof), a thermophilic, acidophilic, amylolytic fungus, for the conversion of potato processing wastes into microbial protein for use as animal feed was studied. The fungus was not inhibited by α-solanine or β-2-chaconine, antimicrobial compounds in potatoes, or by morpholine or cyclohexylamine (additives to steam used in the peeling process) at levels likely to be encountered in this substrate. Mixed effluent from holding tanks at a potato-processing plant contained about 109 bacteria per ml and inhibited fungal growth. The fungus grew well on fresh potato wastes containing up to 5% total carbohydrate and utilized both starch and protein at 45°C and pH 3.75. On potato homogenate medium containing 2% carbohydrate (about 14% fresh potato) supplemented with monoammonium phosphate (0.506 g/liter) and ferric iron (0.1 g/liter), with pH control (at 3.75) and additional nitrogen supplied by the automatic addition of ammonium hydroxide, typical yields were 0.61 g (dry weight) of product and 0.3 g of crude protein per g of carbohydrate supplied. An aerobic, spore-forming bacterium, related to Bacillus brevis, commonly contaminated nonsterilized batch cultures but was destroyed by heating for 15 min at 100°C. PMID:16347277

  5. ChemCell : a particle-based model of protein chemistry and diffusion in microbial cells.

    SciTech Connect

    Plimpton, Steven James; Slepoy, Alexander

    2003-12-01

    Prokaryotic single-cell microbes are the simplest of all self-sufficient living organisms. Yet microbes create and use much of the molecular machinery present in more complex organisms, and the macro-molecules in microbial cells interact in regulatory, metabolic, and signaling pathways that are prototypical of the reaction networks present in all cells. We have developed a simple simulation model of a prokaryotic cell that treats proteins, protein complexes, and other organic molecules as particles which diffuse via Brownian motion and react with nearby particles in accord with chemical rate equations. The code models protein motion and chemistry within an idealized cellular geometry. It has been used to simulate several simple reaction networks and compared to more idealized models which do not include spatial effects. In this report we describe an initial version of the simulation code that was developed with FY03 funding. We discuss the motivation for the model, highlight its underlying equations, and describe simulations of a 3-stage kinase cascade and a portion of the carbon fixation pathway in the Synechococcus microbe.

  6. Degradation of spent craft brewer’s yeast by caprine rumen hyper ammonia-producing bacteria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spent brewer’s yeast has long been included in ruminant diets as a protein supplement. However, modern craft beers often include more hops (Humulus lupulus L.) compounds than traditional recipes. These compounds include alpha and beta-acids, which are antimicrobial to the rumen hyper ammonia-produci...

  7. Ruminococcal cellulosome systems from rumen to human.

    PubMed

    Ben David, Yonit; Dassa, Bareket; Borovok, Ilya; Lamed, Raphael; Koropatkin, Nicole M; Martens, Eric C; White, Bryan A; Bernalier-Donadille, Annick; Duncan, Sylvia H; Flint, Harry J; Bayer, Edward A; Moraïs, Sarah

    2015-09-01

    A cellulolytic fiber-degrading bacterium, Ruminococcus champanellensis, was isolated from human faecal samples, and its genome was recently sequenced. Bioinformatic analysis of the R. champanellensis genome revealed numerous cohesin and dockerin modules, the basic elements of the cellulosome, and manual sequencing of partially sequenced genomic segments revealed two large tandem scaffoldin-coding genes that form part of a gene cluster. Representative R. champanellensis dockerins were tested against putative cohesins, and the results revealed three different cohesin-dockerin binding profiles which implied two major types of cellulosome architectures: (i) an intricate cell-bound system and (ii) a simplistic cell-free system composed of a single cohesin-containing scaffoldin. The cell-bound system can adopt various enzymatic architectures, ranging from a single enzyme to a large enzymatic complex comprising up to 11 enzymes. The variety of cellulosomal components together with adaptor proteins may infer a very tight regulation of its components. The cellulosome system of the human gut bacterium R. champanellensis closely resembles that of the bovine rumen bacterium Ruminococcus flavefaciens. The two species contain orthologous gene clusters comprising fundamental components of cellulosome architecture. Since R. champanellensis is the only human colonic bacterium known to degrade crystalline cellulose, it may thus represent a keystone species in the human gut.

  8. Investigating the effect of two methane-mitigating diets on the rumen microbiome using massively parallel sequencing.

    PubMed

    Ross, E M; Moate, P J; Marett, L; Cocks, B G; Hayes, B J

    2013-09-01

    methane-mitigating diets have altered the rumen microbiome toward naturally low methane-emitting microbial profiles. The contig sequences are predominantly new and include Faecalibacterium spp. The contigs we have identified here are potential biomarkers for low-methane-emitting cattle.

  9. Dietary supplementation of usnic acid, an antimicrobial compound in lichens, does not affect rumen bacterial diversity or density in reindeer.

    PubMed

    Glad, Trine; Barboza, Perry; Mackie, Roderick I; Wright, André-Denis G; Brusetti, Lorenzo; Mathiesen, Svein D; Sundset, Monica A

    2014-06-01

    Reindeer (Rangifer tarandus tarandus) may include large proportions of lichens in their winter diet. These dietary lichens are rich in phenolic secondary compounds, the most well-known being the antimicrobial usnic acid. Previous studies have shown that reindeer host rumen bacteria resistant to usnic acid and that usnic acid is quickly detoxified in their rumen. In the present study, reindeer (n = 3) were sampled before, during, and after usnic acid supplementation to determine the effect on their rumen microbial ecology. Ad libitum intake of usnic acid averaged up to 278 mg/kg body mass. Population densities of rumen bacteria and methanogenic archaea determined by real-time PCR, ranged from 1.36 × 10(9) to 11.8 × 10(9) and 9.0 × 10(5) to 1.35 × 10(8) cells/g wet weight, respectively, and the two populations did not change significantly during usnic acid supplementation (repeated measures ANOVA) or vary significantly between the rumen liquid and particle fraction (paired t test). Rumen bacterial community structure determined by denaturing gradient gel electrophoresis did not change in response to intake of usnic acid. Firmicutes (38.7 %) and Bacteriodetes (27.4 %) were prevalent among the 16S rRNA gene sequences (n = 62) from the DGGE gels, but representatives of the phyla Verrucomicrobia (14.5 %) and Proteobacteria (1.6 %) were also detected. Rapid detoxification of the usnic acid or resistance to usnic acid may explain why the diversity of the dominant bacterial populations and the bacterial density in the reindeer rumen does not change during usnic acid supplementation.

  10. Investigating the effect of two methane-mitigating diets on the rumen microbiome using massively parallel sequencing.

    PubMed

    Ross, E M; Moate, P J; Marett, L; Cocks, B G; Hayes, B J

    2013-09-01

    methane-mitigating diets have altered the rumen microbiome toward naturally low methane-emitting microbial profiles. The contig sequences are predominantly new and include Faecalibacterium spp. The contigs we have identified here are potential biomarkers for low-methane-emitting cattle. PMID:23871375

  11. Microbial fuel cells and microbial ecology: applications in ruminant health and production research.

    PubMed

    Bretschger, Orianna; Osterstock, Jason B; Pinchak, William E; Ishii, Shun'ichi; Nelson, Karen E

    2010-04-01

    Microbial fuel cell (MFC) systems employ the catalytic activity of microbes to produce electricity from the oxidation of organic, and in some cases inorganic, substrates. MFC systems have been primarily explored for their use in bioremediation and bioenergy applications; however, these systems also offer a unique strategy for the cultivation of synergistic microbial communities. It has been hypothesized that the mechanism(s) of microbial electron transfer that enable electricity production in MFCs may be a cooperative strategy within mixed microbial consortia that is associated with, or is an alternative to, interspecies hydrogen (H(2)) transfer. Microbial fermentation processes and methanogenesis in ruminant animals are highly dependent on the consumption and production of H(2)in the rumen. Given the crucial role that H(2) plays in ruminant digestion, it is desirable to understand the microbial relationships that control H(2) partial pressures within the rumen; MFCs may serve as unique tools for studying this complex ecological system. Further, MFC systems offer a novel approach to studying biofilms that form under different redox conditions and may be applied to achieve a greater understanding of how microbial biofilms impact animal health. Here, we present a brief summary of the efforts made towards understanding rumen microbial ecology, microbial biofilms related to animal health, and how MFCs may be further applied in ruminant research.

  12. Microbial Fuel Cells and Microbial Ecology: Applications in Ruminant Health and Production Research

    PubMed Central

    Osterstock, Jason B.; Pinchak, William E.; Ishii, Shun’ichi; Nelson, Karen E.

    2009-01-01

    Microbial fuel cell (MFC) systems employ the catalytic activity of microbes to produce electricity from the oxidation of organic, and in some cases inorganic, substrates. MFC systems have been primarily explored for their use in bioremediation and bioenergy applications; however, these systems also offer a unique strategy for the cultivation of synergistic microbial communities. It has been hypothesized that the mechanism(s) of microbial electron transfer that enable electricity production in MFCs may be a cooperative strategy within mixed microbial consortia that is associated with, or is an alternative to, interspecies hydrogen (H2) transfer. Microbial fermentation processes and methanogenesis in ruminant animals are highly dependent on the consumption and production of H2in the rumen. Given the crucial role that H2 plays in ruminant digestion, it is desirable to understand the microbial relationships that control H2 partial pressures within the rumen; MFCs may serve as unique tools for studying this complex ecological system. Further, MFC systems offer a novel approach to studying biofilms that form under different redox conditions and may be applied to achieve a greater understanding of how microbial biofilms impact animal health. Here, we present a brief summary of the efforts made towards understanding rumen microbial ecology, microbial biofilms related to animal health, and how MFCs may be further applied in ruminant research. PMID:20024685

  13. Potential functional gene diversity involved in methanogenesis and methanogenic community structure in Indian buffalo (Bubalus bubalis) rumen.

    PubMed

    Singh, Krishna M; Patel, Amrutlal K; Shah, Ravi K; Reddy, Bhaskar; Joshi, Chaitanya G

    2015-08-01

    Understanding the methanogen community structure and methanogenesis from Bubalus bubalis in India may be beneficial to methane mitigation. Our current understanding of the microbial processes leading to methane production is incomplete, and further advancement in the knowledge of methanogenesis pathways would provide means to manipulate its emission in the future. In the present study, we evaluated the methanogenic community structure in the rumen as well as their potential genes involved in methanogenesis. The taxonomic and metabolic profiles of methanogens were assessed by shotgun sequencing of rumen metagenome by Ion Torrent semiconductor sequencing. The buffalo rumen contained representative genera of all the families of methanogens. Members of Methanobacteriaceae were found to be dominant, followed by Methanosarcinaceae, Methanococcaceae, Methanocorpusculaceae, and Thermococcaceae. A total of 60 methanogenic genera were detected in buffalo rumen. Methanogens related to the genera Methanobrevibacter, Methanosarcina, Methanococcus, Methanocorpusculum, Methanothermobacter, and Methanosphaera were predominant, representing >70 % of total archaeal sequences. The metagenomic dataset indicated the presence of genes involved in the methanogenesis and acetogenesis pathways, and the main functional genes were those of key enzymes in the methanogenesis. Sequences related to CoB--CoM heterodisulfide reductase, methyl coenzyme M reductase, f420-dependent methylenetetrahydromethanopterin reductase, and formylmethanofuran dehydrogenase were predominant in rumen. In addition, methenyltetrahydrofolate cyclohydrolase, methylenetetrahydrofolate dehydrogenase, 5,10-methylenetetrahydrofolate reductase, and acetyl-coenzyme A synthetase were also recovered. PMID:25663664

  14. Isolation and characterization of novel multifunctional recombinant family 26 glycoside hydrolase from Mehsani buffalo rumen metagenome.

    PubMed

    Patel, Avani B; Patel, Amrutlal K; Shah, Mihir P; Parikh, Ishan K; Joshi, Chaitanya G

    2016-01-01

    Rumen microbiota harbor a diverse set of carbohydrate-active enzymes (CAZymes), which play a crucial role in the degradation of a complex plant polysaccharide thereby providing metabolic energy to the host animals. Earlier, we reported CAZYme analysis from the buffalo rumen metagenome by high throughput shotgun sequencing. Among the various CAZymes, glycoside hydrolase family 26 (GH26) enzymes have a number of industrial applications including in paper, oil, biofuel, food, feed, pharmaceutical, coffee, and detergent industries. Here, we report isolation and characterization of GH26 enzyme from the buffalo rumen metagenome. A novel GH26 gene composed of 1,119 base pairs was successfully amplified using the gene-specific primers inferred based on the contig generated from metagenome sequence assembly and cloned in a pET32a (+) expression vector as an N-terminal histidine tag fusion protein. A novel GH26 protein from an unknown rumen microorganism shared a maximum of 68% identity with the Prevotella ruminicola 23 encoded carbohydrate esterase family 7 and 46% with Bacteroides sp. 2_1_33B encoded mannan endo-1, 4-β-mannosidase. The recombinant GH26-histidine tag fusion protein was expressed in Escherichia coli and purified using Ni-NTA affinity chromatography. The purified enzyme displayed multifunctional activities against various carbohydrate substrates including locust bean gum, beechwood xylan, pectin, and carboxymethyl cellulose suggesting mannanase, xylanase, pectin esterase, and endoglucanase activities, respectively. PMID:25644118

  15. Technical note: Protozoa-specific antibodies raised in sheep plasma bind to their target protozoa in the rumen.

    PubMed

    Williams, Y J; Rea, S M; Popovski, S; Skillman, L C; Wright, A-D G

    2014-12-01

    Binding of IgG antibodies to Entodinium spp. in the rumen of sheep (Ovis aries) was investigated by adding IgG, purified from plasma, directly into the rumen. Plasma IgG was sourced from sheep that had or had not been immunized with a vaccine containing whole fixed Entodinium spp. cells. Ruminal fluid was sampled approximately 2 h after each antibody dosing. Binding of protozoa by a specific antibody was detected using an indirect fluorescent antibody test. An antibody titer in the ruminal fluid was determined by ELISA, and the concentration of ruminal fluid ammonia-N and ruminal pH were also determined. Entodinium spp. and total protozoa from IgG-infused sheep were enumerated by microscopic counts. Two-hourly additions of IgG maintained a low antibody titer in the rumen for 12 h and the binding of the antibody to the rumen protozoa was demonstrated. Increased ammonia-N concentrations and altered ruminal fluid pH patterns indicated that additional fermentation of protein was occurring in the rumen after addition of IgG. No reduction in numbers of Entodinium spp. was observed (P>0.05). Although binding of antibodies to protozoa has been demonstrated in the rumen, it is unclear how much cell death occurred. On the balance of probability, it would appear that the antibody was degraded or partially degraded, and the impact of this on protozoal populations and the measurement of a specific titer is also unclear.

  16. Effects of Moringa oleifera seed extract on rumen fermentation in vitro.

    PubMed

    Hoffmann, E M; Muetzel, S; Becker, K

    2003-02-01

    Moringa oleifera is a pantropical tree of the family Moringaceae. A previously undescribed property of an aqueous extract from the seeds of this plant is the modulation of ruminal fermentation patterns, especially protein degradation, as demonstrated in a short-term batch incubation system. Gas, short chain fatty acids (SCFA) and cellulolytic enzyme activities were determined as general fermentation parameters. A dot blot assay able to directly detect true protein in rumen fluid samples was used to quantify protein degradation. For complex substrates the interpretation of protein degradation profiles was amended by polyacrylamide gel electrophoresis (PAGE) of the samples. When incubated with pure carbohydrates at a concentration of 1 mg ml(-1), the extract reduced microbial degradation of the model protein, bovine serum albumin (BSA), such that its concentration was at least 40% above the control after 12 h of incubation. Total protein degradation was thus delayed by approximately 9 h. When fermented along with wheat straw, leaf protein (Rubisco) was almost entirely protected during 12 h of fermentation. The degradation of soy proteins was retarded by at least 4-6 h, depending on the protein band. There were strong side effects on the fermentation of pure cellulose (SCFA yield-60% after 12 h), whereas cellobiose and starch fermentation were less affected (-18 and -8%, respectively). When the complex substrates were fermented, SCFA yield was reduced by approximately 30% after 12 h. In our work we clearly demonstrate the efficacy of the new substance, which is neither a tannin nor a saponin, in an in vitro system, using pure as well as complex substrates. The properties shown in vitro for the crude extract suggest that it could have a positive effect on the protein metabolism of ruminants under intensive management and that negative side effects can be overcome by an optimized dosage. If the chemical nature of the active substance and its mechanism of action can be

  17. Oxidation-induced unfolding facilitates Myosin cross-linking in myofibrillar protein by microbial transglutaminase.

    PubMed

    Li, Chunqiang; Xiong, Youling L; Chen, Jie

    2012-08-15

    Myofibrillar protein from pork Longissimus muscle was oxidatively stressed for 2 and 24 h at 4 °C with mixed 10 μM FeCl(3)/100 μM ascorbic acid/1, 5, or 10 mM H(2)O(2) (which produces hydroxyl radicals) and then treated with microbial transglutaminase (MTG) (E:S = 1:20) for 2 h at 4 °C. Oxidation induced significant protein structural changes (P < 0.05) as evidenced by suppressed K-ATPase activity, elevated Ca-ATPase activity, increased carbonyl and disulfide contents, and reduced conformational stability, all in a H(2)O(2) dose-dependent manner. The structural alterations, notably with mild oxidation, led to stronger MTG catalysis. More substantial amine reductions (19.8-27.6%) at 1 mM H(2)O(2) occurred as compared to 11.6% in nonoxidized samples (P < 0.05) after MTG treatment. This coincided with more pronounced losses of myosin in oxidized samples (up to 33.2%) as compared to 21.1% in nonoxidized (P < 0.05), which was attributed to glutamine-lysine cross-linking as suggested by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID:22809283

  18. Catabolism of tritiated thymidine by aquatic microbial communities and incorporation of tritium into RNA and protein

    SciTech Connect

    Brittain, A.M.; Karl, D.M. )

    1990-05-01

    The incorporation of tritiated thymidine by five microbial ecosystems and the distribution of tritium into DNA, RNA, and protein were determined. Nonspecific labeling was greatest in sediment samples, for which {>=}95% of the tritium was recovered with the RNA and protein fractions. The percentage of tritium recovered in the DNA fraction ranged from 15 to 38% of the total labeled macromolecules recovered. Nonspecific labeling was independent of both incubation time and thymidine concentration over very wide ranges. We also evaluated the specificity of (2-{sup 3}H) adenine incorporation into adenylate residues in both RNA and DNA in parallel with the ({sup 3}H) thymidine experiments and compared the degree of nonspecific labeling by ({sup 3}H) adenine with that derived from ({sup 3}H)thymidine. Rapid catabolism of tritiated thymidine was evaluated by determining the disappearance of tritiated thymidine from the incubation medium and the appearance of degradation products. Degradation product formation, including that of both volatile and nonvolatile compounds, was much greater than the rate of incorporation of tritium into stable macromolecules. The standard degradation pathway for thymidine coupled with utilization of Krebs cycle intermediates for the biosynthesis of amino acids, purines, and pyrimidines readily accounts for the observed nonspecific labeling in environmental samples.

  19. Unconventional microbial systems for the cost-efficient production of high-quality protein therapeutics.

    PubMed

    Corchero, José Luis; Gasser, Brigitte; Resina, David; Smith, Wesley; Parrilli, Ermenegilda; Vázquez, Felícitas; Abasolo, Ibane; Giuliani, Maria; Jäntti, Jussi; Ferrer, Pau; Saloheimo, Markku; Mattanovich, Diethard; Schwartz, Simó; Tutino, Maria Luisa; Villaverde, Antonio

    2013-01-01

    Both conventional and innovative biomedical approaches require cost-effective protein drugs with high therapeutic potency, improved bioavailability, biocompatibility, stability and pharmacokinetics. The growing longevity of the human population, the increasing incidence and prevalence of age-related diseases and the better comprehension of genetic-linked disorders prompt to develop natural and engineered drugs addressed to fulfill emerging therapeutic demands. Conventional microbial systems have been for long time exploited to produce biotherapeutics, competing with animal cells due to easier operation and lower process costs. However, both biological platforms exhibit important drawbacks (mainly associated to intracellular retention of the product, lack of post-translational modifications and conformational stresses), that cannot be overcome through further strain optimization merely due to physiological constraints. The metabolic diversity among microorganisms offers a spectrum of unconventional hosts, that, being able to bypass some of these weaknesses, are under progressive incorporation into production pipelines. In this review we describe the main biological traits and potentials of emerging bacterial, yeast, fungal and microalgae systems, by comparing selected leading species with well established conventional organisms with a long run in protein drug production.

  20. Role of rumen butyrate in regulation of nitrogen utilization and urea nitrogen kinetics in growing sheep.

    PubMed

    Agarwal, U; Hu, Q; Baldwin, R L; Bequette, B J

    2015-05-01

    Butyrate, a major rumen VFA, has been indirectly linked to enhancement of urea recycling on the basis of increased expression of urea transporter in the rumen epithelia of steers fed a rumen butyrate-enhancing diet. Two studies were conducted to quantify the effect of elevated rumen butyrate concentrations on N balance, urea kinetics and rumen epithelial proliferation. Wether sheep (n= 4), fitted with a rumen cannula, were fed a pelleted ration (∼165 g CP/kg DM, 10.3 MJ ME/kg DM) at 1.8 × ME requirement. In Exp. 1, sheep were infused intraruminally with either an electrolyte buffer solution (Con-Buf) or butyrate dissolved in the buffer solution (But-Buf) during 8-d periods in a balanced crossover design. In Exp. 2, sheep were infused intraruminally with either sodium acetate (Na-Ac) or sodium butyrate (Na-But) for 9 d. All solutions were adjusted to pH 6.8 and 8.0 in Exp. 1 and 2, respectively, and VFA were infused at 10% of ME intake. [15N2] urea was continuously infused intravenously for the last 5 d of each period, and total urine and feces were collected. In Exp. 1, 2H5-phenylalanine was continuously infused intravenously over the last 12 h, after which a biopsy from the rumen papillae was taken for measurement of fractional protein synthesis rate (FSR). Butyrate infusion treatments increased (P = 0.1 in Exp. 1; P < 0.05 in Exp. 2) the proportion of rumen butyrate, and acetate infusion increased (P < 0.05) rumen acetate. All animals were in positive N balance (4.2 g N/d in Exp. 1; 7.0 g N/d in Exp. 2), but no difference in N retention was observed between treatments. In Exp. 2, urea entry (synthesis) rate was reduced ( < 0.05) by Na-But compared with the Na-Ac control. In Exp. 1, although But-Buf infusion increased the FSR of rumen papillae (35.3% ± 1.08%/d vs. 28.7% ± 1.08%/d; P < 0.05), urea kinetics were not altered by But-Buf compared with Con-Buf. These studies are the first to directly assess the role of butyrate in urea recycling and its effects on

  1. Metagenomic analysis of buffalo rumen microbiome: Effect of roughage diet on Dormancy and Sporulation genes

    PubMed Central

    Singh, K.M.; Reddy, B.; Patel, A.K.; Panchasara, H.; Parmar, N.; Patel, A.B.; Shah, T.M.; Bhatt, V.D.; Joshi, C.G.

    2014-01-01

    Buffalo rumen microbiome experiences a variety of diet stress and represents reservoir of Dormancy and Sporulation genes. However, the information on genomic responses to such conditions is very limited. The Ion Torrent PGM next generation sequencing technology was used to characterize general microbial diversity and the repertoire of microbial genes present, including genes associated with Dormancy and Sporulation in Mehsani buffalo rumen metagenome. The research findings revealed the abundance of bacteria at the domain level and presence of Dormancy and Sporulation genes which were predominantly associated with the Clostridia and Bacilli taxa belonging to the phyla Firmicutes. Genes associated with Sporulation cluster and Sporulation orphans were increased from 50% to 100% roughage treatment, thereby promoting sporulation all along the treatments. The spore germination is observed to be the highest in the 75% roughage treatment both in the liquid and solid rumen fraction samples with respect to the decrease in the values of the genes associated with spore core dehydration, thereby facilitating spore core hydration which is necessary for spore germination. PMID:25606408

  2. Metagenomic analysis of buffalo rumen microbiome: Effect of roughage diet on Dormancy and Sporulation genes.

    PubMed

    Singh, K M; Reddy, B; Patel, A K; Panchasara, H; Parmar, N; Patel, A B; Shah, T M; Bhatt, V D; Joshi, C G

    2014-12-01

    Buffalo rumen microbiome experiences a variety of diet stress and represents reservoir of Dormancy and Sporulation genes. However, the information on genomic responses to such conditions is very limited. The Ion Torrent PGM next generation sequencing technology was used to characterize general microbial diversity and the repertoire of microbial genes present, including genes associated with Dormancy and Sporulation in Mehsani buffalo rumen metagenome. The research findings revealed the abundance of bacteria at the domain level and presence of Dormancy and Sporulation genes which were predominantly associated with the Clostridia and Bacilli taxa belonging to the phyla Firmicutes. Genes associated with Sporulation cluster and Sporulation orphans were increased from 50% to 100% roughage treatment, thereby promoting sporulation all along the treatments. The spore germination is observed to be the highest in the 75% roughage treatment both in the liquid and solid rumen fraction samples with respect to the decrease in the values of the genes associated with spore core dehydration, thereby facilitating spore core hydration which is necessary for spore germination. PMID:25606408

  3. Prepartum and Postpartum Rumen Fluid Microbiomes: Characterization and Correlation with Production Traits in Dairy Cows

    PubMed Central

    Lima, Fabio S.; Oikonomou, Georgios; Lima, Svetlana F.; Bicalho, Marcela L. S.; Ganda, Erika K.; de Oliveira Filho, Jose C.; Lorenzo, Gustavo; Trojacanec, Plamen

    2014-01-01

    Microbes present in the rumen of dairy cows are essential for degradation of cellulosic and nonstructural carbohydrates of plant origin. The prepartum and postpartum diets of high-producing dairy cows are substantially different, but in what ways the rumen microbiome changes in response and how those changes may influence production traits are not well elucidated. Here, we sequenced the 16S and 18S rRNA genes using the MiSeq platform to characterize the prepartum and postpartum rumen fluid microbiomes in 115 high-producing dairy cows, including both primiparous and multiparous animals. Discriminant analysis identified differences between the microbiomes of prepartum and postpartum samples and between primiparous and multiparous cows. 18S rRNA sequencing revealed an overwhelming dominance of the protozoan class Litostomatea, with over 90% of the eukaryotic microbial population belonging to that group. Additionally, fungi were relatively more prevalent and Litostomatea relatively less prevalent in prepartum samples than in postpartum ones. The core rumen microbiome (common to all samples) consisted of 64 bacterial taxa, of which members of the genus Prevotella were the most prevalent. The Chao1 richness index was greater for prepartum multiparous cows than for postpartum multiparous cows. Multivariable models identified bacterial taxa associated with increased or reduced milk production, and general linear models revealed that a metagenomically based prediction of productivity is highly associated with production of actual milk and milk components. In conclusion, the structure of the rumen fluid microbiome shifts between the prepartum and first-week postpartum periods, and its profile within the context of this study could be used to accurately predict production traits. PMID:25501481

  4. Rumen motility during induced hyper- and hypocalcaemia.

    PubMed

    Jørgensen, R J; Nyengaard, N R; Hara, S; Enemark, J M; Andersen, P H

    1998-01-01

    Rumen motility was recorded on an experimental cow by means of telemetric signal transfer from strain gauge force transducers fixed surgically on the peritoneal surface of the rumen wall in the left flank. The normocalcaemic cow was given a standard milk fever treatment with calcium borogluconate (400 ml with 14 mg Ca/ml) intravenously. Transient clinical signs were: decreased rumination, muscle ticks, salivation and a heart rate reduction of 20%. Rectal temperature remained unaltered. Frequency of rumen contractions was reduced up to 40% whereas amplitude of contractions did not deviate from baseline values. Hypocalcaemia was induced in a second experiment by iv infusion of Na2EDTA. At 0.60 mmol/l ionized blood calcium periods of no motility were recorded whereas inactivity of rumen activity was persistent at 0.55 mmol/l ionized blood calcium. The cow went down at 0.45-0.48 mmol/l ionized blood calcium at which point the heart rate was increased by 40%. The high sensitivity of the method employed allowed the conclusion that already at a concentration of ionized blood calcium at 1.0 mmol/l both frequency and amplitude of rumen contractions decreased rapidly although eating behaviour and rumination appeared unaffected during the short term observation periods. Implications of this finding towards health and production in transition cows are discussed.

  5. Supplementing barley or rapeseed meal to dairy cows fed grass-red clover silage: II. Amino acid profile of microbial fractions.

    PubMed

    Korhonen, M; Ahvenjärvi, S; Vanhatalo, A; Huhtanen, P

    2002-08-01

    Four ruminally cannulated dairy cows were used to examine the effect of diet on the AA composition of rumen bacteria and protozoa, and the flow of microbial and nonmicrobial AA entering the omasal canal. Cows were offered grass-red clover silage alone, or that supplemented with 5.1 kg DM of barley, 1.9 kg DM of rapeseed meal, or 5.1 kg DM of barley and 1.9 kg DM of rapeseed meal according to a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. During the first 10 d of each period, cows had free access to silage and, thereafter intake was restricted to 95% of ad libitum intake. Postruminal digesta flow was assessed using the omasal canal sampling technique in combination with a triple marker method. Liquid- (LAB) and particle- (PAB) associated bacteria were isolated from digesta in the reticulorumen and protozoa from digesta entering the omasal canal. Microbial protein flow was determined using 15N as a microbial marker. Flows of AA entering the omasal canal were similar in cows fed silage diets supplemented with barley or rapeseed meal. However, rapeseed meal increased nonmicrobial AA flow while barley increased the flow of AA associated with LAB and protozoa. Diet had negligible effects on the AA profile of microbial fractions. Comparison of AA profiles across diets indicated differences between LAB and PAB for 10 out of 17 AA measured. Rumen bacteria and protozoa were found to be different for 14 out of 15 AA measured. For grass silage-based diets, energy and protein supplementations appear to alter postruminal AA supply through modifications in the proportionate contribution of microbial and nonmicrobial pools to total protein flow rather than as a direct result of changes in the AA profile of microbial protein.

  6. Temporal dynamics of fibrolytic and methanogenic rumen microorganisms during in situ incubation of switchgrass determined by 16S rRNA gene profiling.

    PubMed

    Piao, Hailan; Lachman, Medora; Malfatti, Stephanie; Sczyrba, Alexander; Knierim, Bernhard; Auer, Manfred; Tringe, Susannah G; Mackie, Roderick I; Yeoman, Carl J; Hess, Matthias

    2014-01-01

    The rumen microbial ecosystem is known for its biomass-degrading and methane-producing phenotype. Fermentation of recalcitrant plant material, comprised of a multitude of interwoven fibers, necessitates the synergistic activity of diverse microbial taxonomic groups that inhabit the anaerobic rumen ecosystem. Although interspecies hydrogen (H2) transfer, a process during which bacterially generated H2 is transferred to methanogenic Archaea, has obtained significant attention over the last decades, the temporal variation of the different taxa involved in in situ biomass-degradation, H2 transfer and the methanogenesis process remains to be established. Here we investigated the temporal succession of microbial taxa and its effect on fiber composition during rumen incubation using 16S rRNA amplicon sequencing. Switchgrass filled nylon bags were placed in the rumen of a cannulated cow and collected at nine time points for DNA extraction and 16S pyrotag profiling. The microbial community colonizing the air-dried and non-incubated (0 h) switchgrass was dominated by members of the Bacilli (recruiting 63% of the pyrotag reads). During in situ incubation of the switchgrass, two major shifts in the community composition were observed: Bacilli were replaced within 30 min by members belonging to the Bacteroidia and Clostridia, which recruited 34 and 25% of the 16S rRNA reads generated, respectively. A second significant shift was observed after 16 h of rumen incubation, when members of the Spirochaetes and Fibrobacteria classes became more abundant in the fiber-adherent community. During the first 30 min of rumen incubation ~13% of the switchgrass dry matter was degraded, whereas little biomass degradation appeared to have occurred between 30 min and 4 h after the switchgrass was placed in the rumen. Interestingly, methanogenic members of the Euryarchaeota (i.e., Methanobacteria) increased up to 3-fold during this period of reduced biomass-degradation, with peak abundance just

  7. Temporal dynamics of fibrolytic and methanogenic rumen microorganisms during in situ incubation of switchgrass determined by 16S rRNA gene profiling.

    PubMed

    Piao, Hailan; Lachman, Medora; Malfatti, Stephanie; Sczyrba, Alexander; Knierim, Bernhard; Auer, Manfred; Tringe, Susannah G; Mackie, Roderick I; Yeoman, Carl J; Hess, Matthias

    2014-01-01

    The rumen microbial ecosystem is known for its biomass-degrading and methane-producing phenotype. Fermentation of recalcitrant plant material, comprised of a multitude of interwoven fibers, necessitates the synergistic activity of diverse microbial taxonomic groups that inhabit the anaerobic rumen ecosystem. Although interspecies hydrogen (H2) transfer, a process during which bacterially generated H2 is transferred to methanogenic Archaea, has obtained significant attention over the last decades, the temporal variation of the different taxa involved in in situ biomass-degradation, H2 transfer and the methanogenesis process remains to be established. Here we investigated the temporal succession of microbial taxa and its effect on fiber composition during rumen incubation using 16S rRNA amplicon sequencing. Switchgrass filled nylon bags were placed in the rumen of a cannulated cow and collected at nine time points for DNA extraction and 16S pyrotag profiling. The microbial community colonizing the air-dried and non-incubated (0 h) switchgrass was dominated by members of the Bacilli (recruiting 63% of the pyrotag reads). During in situ incubation of the switchgrass, two major shifts in the community composition were observed: Bacilli were replaced within 30 min by members belonging to the Bacteroidia and Clostridia, which recruited 34 and 25% of the 16S rRNA reads generated, respectively. A second significant shift was observed after 16 h of rumen incubation, when members of the Spirochaetes and Fibrobacteria classes became more abundant in the fiber-adherent community. During the first 30 min of rumen incubation ~13% of the switchgrass dry matter was degraded, whereas little biomass degradation appeared to have occurred between 30 min and 4 h after the switchgrass was placed in the rumen. Interestingly, methanogenic members of the Euryarchaeota (i.e., Methanobacteria) increased up to 3-fold during this period of reduced biomass-degradation, with peak abundance just

  8. Temporal dynamics of fibrolytic and methanogenic rumen microorganisms during in situ incubation of switchgrass determined by 16S rRNA gene profiling

    PubMed Central

    Piao, Hailan; Lachman, Medora; Malfatti, Stephanie; Sczyrba, Alexander; Knierim, Bernhard; Auer, Manfred; Tringe, Susannah G.; Mackie, Roderick I.; Yeoman, Carl J.; Hess, Matthias

    2014-01-01

    The rumen microbial ecosystem is known for its biomass-degrading and methane-producing phenotype. Fermentation of recalcitrant plant material, comprised of a multitude of interwoven fibers, necessitates the synergistic activity of diverse microbial taxonomic groups that inhabit the anaerobic rumen ecosystem. Although interspecies hydrogen (H2) transfer, a process during which bacterially generated H2 is transferred to methanogenic Archaea, has obtained significant attention over the last decades, the temporal variation of the different taxa involved in in situ biomass-degradation, H2 transfer and the methanogenesis process remains to be established. Here we investigated the temporal succession of microbial taxa and its effect on fiber composition during rumen incubation using 16S rRNA amplicon sequencing. Switchgrass filled nylon bags were placed in the rumen of a cannulated cow and collected at nine time points for DNA extraction and 16S pyrotag profiling. The microbial community colonizing the air-dried and non-incubated (0 h) switchgrass was dominated by members of the Bacilli (recruiting 63% of the pyrotag reads). During in situ incubation of the switchgrass, two major shifts in the community composition were observed: Bacilli were replaced within 30 min by members belonging to the Bacteroidia and Clostridia, which recruited 34 and 25% of the 16S rRNA reads generated, respectively. A second significant shift was observed after 16 h of rumen incubation, when members of the Spirochaetes and Fibrobacteria classes became more abundant in the fiber-adherent community. During the first 30 min of rumen incubation ~13% of the switchgrass dry matter was degraded, whereas little biomass degradation appeared to have occurred between 30 min and 4 h after the switchgrass was placed in the rumen. Interestingly, methanogenic members of the Euryarchaeota (i.e., Methanobacteria) increased up to 3-fold during this period of reduced biomass-degradation, with peak abundance just

  9. Associative patterns among anaerobic fungi, methanogenic archaea, and bacterial communities in response to changes in diet and age in the rumen of dairy cows.

    PubMed

    Kumar, Sanjay; Indugu, Nagaraju; Vecchiarelli, Bonnie; Pitta, Dipti W

    2015-01-01

    The rumen microbiome represents a complex microbial genetic web where bacteria, anaerobic rumen fungi (ARF), protozoa and archaea work in harmony contributing to the health and productivity of ruminants. We hypothesized that the rumen microbiome shifts as the dairy cow advances in lactations and these microbial changes may contribute to differences in productivity between primiparous (first lactation) and multiparous (≥second lactation) cows. To this end, we investigated shifts in the ruminal ARF and methanogenic communities in both primiparous (n = 5) and multiparous (n = 5) cows as they transitioned from a high forage to a high grain diet upon initiation of lactation. A total of 20 rumen samples were extracted for genomic DNA, amplified using archaeal and fungal specific primers, sequenced on a 454 platform and analyzed using QIIME. Community comparisons (Bray-Curtis index) revealed the effect of diet (P < 0.01) on ARF composition, while archaeal communities differed between primiparous and multiparous cows (P < 0.05). Among ARF, several lineages were unclassified, however, phylum Neocallimastigomycota showed the presence of three known genera. Abundance of Cyllamyces and Caecomyces shifted with diet, whereas Orpinomyces was influenced by both diet and age. Methanobrevibacter constituted the most dominant archaeal genus across all samples. Co-occurrence analysis incorporating taxa from bacteria, ARF and archaea revealed syntrophic interactions both within and between microbial domains in response to change in diet as well as age of dairy cows. Notably, these interactions were numerous and complex in multiparous cows, supporting our hypothesis that the rumen microbiome also matures with age to sustain the growing metabolic needs of the host. This study provides a broader picture of the ARF and methanogenic populations in the rumen of dairy cows and their co-occurrence implicates specific relationships between different microbial domains in response to diet and

  10. Associative patterns among anaerobic fungi, methanogenic archaea, and bacterial communities in response to changes in diet and age in the rumen of dairy cows

    PubMed Central

    Kumar, Sanjay; Indugu, Nagaraju; Vecchiarelli, Bonnie; Pitta, Dipti W.

    2015-01-01

    The rumen microbiome represents a complex microbial genetic web where bacteria, anaerobic rumen fungi (ARF), protozoa and archaea work in harmony contributing to the health and productivity of ruminants. We hypothesized that the rumen microbiome shifts as the dairy cow advances in lactations and these microbial changes may contribute to differences in productivity between primiparous (first lactation) and multiparous (≥second lactation) cows. To this end, we investigated shifts in the ruminal ARF and methanogenic communities in both primiparous (n = 5) and multiparous (n = 5) cows as they transitioned from a high forage to a high grain diet upon initiation of lactation. A total of 20 rumen samples were extracted for genomic DNA, amplified using archaeal and fungal specific primers, sequenced on a 454 platform and analyzed using QIIME. Community comparisons (Bray–Curtis index) revealed the effect of diet (P < 0.01) on ARF composition, while archaeal communities differed between primiparous and multiparous cows (P < 0.05). Among ARF, several lineages were unclassified, however, phylum Neocallimastigomycota showed the presence of three known genera. Abundance of Cyllamyces and Caecomyces shifted with diet, whereas Orpinomyces was influenced by both diet and age. Methanobrevibacter constituted the most dominant archaeal genus across all samples. Co-occurrence analysis incorporating taxa from bacteria, ARF and archaea revealed syntrophic interactions both within and between microbial domains in response to change in diet as well as age of dairy cows. Notably, these interactions were numerous and complex in multiparous cows, supporting our hypothesis that the rumen microbiome also matures with age to sustain the growing metabolic needs of the host. This study provides a broader picture of the ARF and methanogenic populations in the rumen of dairy cows and their co-occurrence implicates specific relationships between different microbial domains in response to diet and

  11. Ultrasonographic Examination of the Rumen in Healthy Cows

    PubMed Central

    Imran, Sheikh; Kumar, Adarsh; Tyagi, S. P.; Kumar, Amit; Sharma, Shivali

    2011-01-01

    10 healthy Indian Jersey/Red Sindhi crossbred nonpregnant cows were subjected to transabdominal ultrasonography to develop baseline topographical data of the rumen. The wall of the rumen could be identified as a thick echogenic line adjacent to the left abdominal wall from left flank to 8th intercostal space. The motility pattern of rumen was characterized by approximately 1 contraction every minute. The mean amplitude of the ruminal contraction was 3.2 cm. Ultrasonography of the rumen in healthy cows is a useful adjunct to the noninvasive diagnostic investigation of the rumen. PMID:21547228

  12. [Effects of organic fish protein liquid fertilizer on enzyme activities and microbial biomass C and N in a silt soil].

    PubMed

    Wei, Xiu-Li; Lei, Ping; Shi, Wei-Yong

    2010-08-01

    By the method of thermostatic culture, this paper studied the effects of different application rates (0.5, 1.5, and 2.5 ml x kg(-1)) of organic fish protein liquid fertilizer on the enzyme activities and microbial biomass C and N in a silt soil, and the relationships between these parameters and soil nutrient contents. Under the application of the liquid fertilizer, soil pH varied in the range of 7.07-7.31, but had no significant difference from the control. With the increasing application rate of the liquid fertilizer, the activities of soil phosphatase, urease, and protease, as well as the soil biomass C and N, all increased significantly, and the increment was 127, 190 and 196%, 39.81, 78.06 and 173.24%, 56.37, 108.29 and 199.98%, 167, 395 and 474%, and 121, 243 and 406%, respectively, compared with the control. The peak time of the soil urease and protease activities and microbial biomass C and N differed with the fertilization treatments. Soil phosphase, urease, and protease activities and microbial biomass C and N were significantly positively correlated with soil nutrient contents, suggesting that applying organic fish protein liquid fertilizer to silt soil could improve soil microbial growth and enzyme activities, and accordingly, promote the decomposition and transformation of soil organic matter and the release of soil available nutrient elements.

  13. Rumen function in vivo and in vitro in sheep fed Leucaena leucocephala.

    PubMed

    Barros-Rodríguez, Marcos Antonio; Solorio-Sánchez, Francisco Javier; Sandoval-Castro, Carlos Alfredo; Klieve, Athol; Rojas-Herrera, Rafael Antonio; Briceño-Poot, Eduardo Gaspar; Ku-Vera, Juan Carlos

    2015-04-01

    The effect of Leucaena leucocephala inclusion in sheep diets upon rumen function was evaluated. Nine Pelibuey sheep, 32.6 ± 5.33 kg live weight (LW), fitted with rumen cannula were used. A complete randomized block design was employed. Two experimental periods of 60 days each, with 60-day intervals between them, were used. Experimental treatments were as follows (n = 6): T1 (control), 100 % Pennisetum purpureum grass; T2, 20 % L. leucocephala + 80 % P. purpureum; T3, 40 % L. leucocephala + 60 % P. purpureum. In situ rumen neutral detergent fiber (aNDF) and crude protein (CP) degradation, dry matter intake (DMI), volatile fatty acids (VFA) production, estimated methane (CH4) yield, rumen pH, ammonia nitrogen (N-NH3), and protozoa counts were measured. The aNDF in situ rumen degradation of P. purpureum and leucaena was higher (P < 0.05) in T2 and T3. Leucaena CP degradation was higher in T2 and T3 but for P. purpureum it was only significantly higher in T3. Leucaena aNDF and CP degradation rate (c) was 50 % higher (P < 0.05) in T2 and T3, but only higher in T3 for P. purpureum. Voluntary intake and rumen (N-NH3) was higher in T2 and T3 (P = 0.0001, P = 0.005, respectively). Molar VFA proportions were similar for all treatments (P > 0.05). Protozoa counts and in vitro gas production (48 h) were lower in T2 and T3 (P < 0.05, P < 0.0001). Estimated methane yield (mol CH4/day) was higher in sheep fed leucaena (P < 0.0001). However, CH4 yield relative to animal performance (mol CH4/g LW gain) was lower in T2 and T3 (P < 0.0001). In summary, these results indicate that including L. leucocephala in sheep diets did not modify rumen fermentation pattern (same VFA ratios) nor reduce the amount of CH4 per unit of DMI (mol CH4/g DMI). However, leucaena inclusion does increase rumen N-NH3, aNDF and CP digestibility, and voluntary intake. PMID:25764346

  14. [Extracorporeal perfusion of the sheep rumen].

    PubMed

    Leng, L; Bajo, M; Várady, J; Szányiová, M

    1977-06-01

    We constructed a modified perfusion apparatus and elaborated a method of extracorporal perfusion of the rumen of sheep. As perfusates we used the bovine plasma diluted in a ratio of 1:1 of an isotonic sodium chloride (NaCl) solution and the whole autologous blood. Transaminases GOT and GPT, ammonia and pH were determined in the perfusate. The different perfusions were evaluated according to previously determined perfusion conditions and criteria. A subject for discussion is the question of suitability of the parameters under examination for judging the state of the perfused organ. The described method is suitable for the study of metabolical processes in the rumen wal.

  15. Ultrasonography of the rumen of dairy cows

    PubMed Central

    2013-01-01

    Background This study describes the ultrasonographic findings of the rumen in 45 healthy dairy cows. Results The cows were scanned on both sides using a 5.0 MHz transducer. The dorsal visible margin of the rumen ran parallel to the lung from cranioventral to caudodorsal. It was furthest from the dorsal midline at the 9th intercostal space (48.3 ± 9.24 cm) and closest at the 12th intercostal space (22.4 ± 3.27 cm). The longitudinal groove, which could be clearly identified at all examination sites because it appeared as a triangular notch, formed the ventral margin of the dorsal sac of the rumen. The dorsal sac of the rumen was largest at the caudal flank (40.3 ± 6.33 cm), where it was adjacent to the abdominal wall. The ventral sac of the rumen extended across the ventral midline into the right hemiabdomen and its ventral margin had a largely horizontal craniocaudal course. The height of the ventral sac of the rumen exceeded that of the dorsal sac at all examination sites; the maximum height was measured at the 12th intercostal space (62.6 ± 9.53 cm). The dorsal gas cap, characterised ultrasonographically by typical reverberation artifacts, was visible in all cows from the 12th intercostal space to the caudal flank. It was largest at the 12th intercostal space (20.5 ± 7.03 cm). The transition from the gas cap to the fibre mat was marked by the abrupt cessation of the reverberation artifacts. It was not possible to differentiate a fibre mat and a ventral fluid phase. The rumen could be imaged from the right side in 21 cows (47%). Conclusions Ultrasonography is well suited for the detailed examination of the rumen of cows. The reference values obtained from this study add to the diagnostic tools that are available for the assessment of bovine patients. PMID:23497545

  16. Rumen papillae morphology of beef steers relative to gain and feed intake and the association of volatile fatty acids with kallikrein gene expression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Feed costs are the most expensive input in beef production. Improvement in the feed efficiency of beef cattle would lower feed inputs and reduce the cost of production. The rumen epithelium is responsible for absorption and metabolism of nutrients and microbial by-products, and may play a significan...

  17. Surface enlargement factor of rumen papillae and expression of genes involved in dermatological diseases and their relationships with feed intake in beef steers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Feed costs make up the largest portion of the total cost to produce beef. One way to reduce this cost, thereby increasing profitability of beef production, is to improve feed efficiency. The rumen is responsible for digestion and absorption of nutrients and microbial by-products and may play a sig...

  18. Epitopes of Microbial and Human Heat Shock Protein 60 and Their Recognition in Myalgic Encephalomyelitis

    PubMed Central

    Elfaitouri, Amal; Herrmann, Björn; Bölin-Wiener, Agnes; Wang, Yilin; Gottfries, Carl-Gerhard; Zachrisson, Olof; Pipkorn, Rϋdiger; Rönnblom, Lars; Blomberg, Jonas

    2013-01-01

    Myalgic encephalomyelitis (ME, also called Chronic Fatigue Syndrome), a common disease with chronic fatigability, cognitive dysfunction and myalgia of unknown etiology, often starts with an infection. The chaperonin human heat shock protein 60 (HSP60) occurs in mitochondria and in bacteria, is highly conserved, antigenic and a major autoantigen. The anti-HSP60 humoral (IgG and IgM) immune response was studied in 69 ME patients and 76 blood donors (BD) (the Training set) with recombinant human and E coli HSP60, and 136 30-mer overlapping and targeted peptides from HSP60 of humans, Chlamydia, Mycoplasma and 26 other species in a multiplex suspension array. Peptides from HSP60 helix I had a chaperonin-like activity, but these and other HSP60 peptides also bound IgG and IgM with an ME preference, theoretically indicating a competition between HSP60 function and antibody binding. A HSP60-based panel of 25 antigens was selected. When evaluated with 61 other ME and 399 non-ME samples (331 BD, 20 Multiple Sclerosis and 48 Systemic Lupus Erythematosus patients), a peptide from Chlamydia pneumoniae HSP60 detected IgM in 15 of 61 (24%) of ME, and in 1 of 399 non-ME at a high cutoff (p<0.0001). IgM to specific cross-reactive epitopes of human and microbial HSP60 occurs in a subset of ME, compatible with infection-induced autoimmunity. PMID:24312270

  19. Epitopes of microbial and human heat shock protein 60 and their recognition in myalgic encephalomyelitis.

    PubMed

    Elfaitouri, Amal; Herrmann, Björn; Bölin-Wiener, Agnes; Wang, Yilin; Gottfries, Carl-Gerhard; Zachrisson, Olof; Pipkorn, Rϋdiger; Rönnblom, Lars; Blomberg, Jonas

    2013-01-01

    Myalgic encephalomyelitis (ME, also called Chronic Fatigue Syndrome), a common disease with chronic fatigability, cognitive dysfunction and myalgia of unknown etiology, often starts with an infection. The chaperonin human heat shock protein 60 (HSP60) occurs in mitochondria and in bacteria, is highly conserved, antigenic and a major autoantigen. The anti-HSP60 humoral (IgG and IgM) immune response was studied in 69 ME patients and 76 blood donors (BD) (the Training set) with recombinant human and E coli HSP60, and 136 30-mer overlapping and targeted peptides from HSP60 of humans, Chlamydia, Mycoplasma and 26 other species in a multiplex suspension array. Peptides from HSP60 helix I had a chaperonin-like activity, but these and other HSP60 peptides also bound IgG and IgM with an ME preference, theoretically indicating a competition between HSP60 function and antibody binding. A HSP60-based panel of 25 antigens was selected. When evaluated with 61 other ME and 399 non-ME samples (331 BD, 20 Multiple Sclerosis and 48 Systemic Lupus Erythematosus patients), a peptide from Chlamydia pneumoniae HSP60 detected IgM in 15 of 61 (24%) of ME, and in 1 of 399 non-ME at a high cutoff (p<0.0001). IgM to specific cross-reactive epitopes of human and microbial HSP60 occurs in a subset of ME, compatible with infection-induced autoimmunity.

  20. Advancing understanding of microbial bioenergy conversion processes by activity-based protein profiling

    DOE PAGESBeta

    Liu, Yun; Fredrickson, James K.; Sadler, Natalie C.; Nandhikonda, Premchendar; Smith, Richard D.; Wright, Aaron T.

    2015-09-25

    Here, the development of renewable biofuels is a global priority, but success will require novel technologies that greatly improve our understanding of microbial systems biology. An approach with great promise in enabling functional characterization of microbes is activity-based protein profiling (ABPP), which employs chemical probes to directly measure enzyme function in discrete enzyme classes in vivo and/or in vitro, thereby facilitating the rapid discovery of new biocatalysts and enabling much improved biofuel production platforms. We review general design strategies in ABPP, and highlight recent advances that are or could be pivotal to biofuels processes including applications of ABPP to cellulosicmore » bioethanol, biodiesel, and phototrophic production of hydrocarbons. We also examine the key challenges and opportunities of ABPP in renewable biofuels research. The integration of ABPP with molecular and systems biology approaches will shed new insight on the catalytic and regulatory mechanisms of functional enzymes and their synergistic effects in the field of biofuels production.« less

  1. Effects of feeding corn silage inoculated with microbial additives on the ruminal fermentation, microbial protein yield, and growth performance of lambs.

    PubMed

    Basso, F C; Adesogan, A T; Lara, E C; Rabelo, C H S; Berchielli, T T; Teixeira, I A M A; Siqueira, G R; Reis, R A

    2014-12-01

    This study aimed to examine the effects of feeding corn silage inoculated without or with either Lactobacillus buchneri (LB) alone or a combination of LB and Lactobacillus plantarum (LBLP) on the apparent digestibility, ruminal fermentation, microbial protein synthesis, and growth performance of lambs. Thirty Santa Inês×Dorper crossbred intact males lambs weighing 20.4±3.8 kg were blocked by weight into 10 groups. Lambs in each group were randomly assigned to 1 of the following 3 dietary treatments: untreated (Control), LB, and LBLP silage. Lambs were fed experimental diets for 61 d. The apparent digestibility was indirectly estimated from indigestible NDF measured on d 57 to 59. Spot urine samples were collected from all animals on d 59 to estimate microbial protein synthesis. Lambs were slaughtered for carcass evaluation on d 61 when they weighed 32.4±5.2 kg. Six additional ruminally cannulated Santa Inês×Dorper crossbred wethers weighing 40.5±1.8 kg were used to examine dietary effects on ruminal fermentation. Average daily gain was increased when lambs were fed LBLP silage (P<0.05) but not LB silage. The LBLP silage had the highest (P<0.05) lactic acid concentration and both inoculated silages had greater acetic acid concentrations than the Control silage (P<0.05). Inoculation of corn silage increased intakes of DM, OM, CP, NDF, total carbohydrate (CHO), and GE by the lambs but decreased digestibility of DM, OM, CP, total and nonstructural carbohydrates, and concentration of GE and ME. (P<0.05). Nevertheless, lambs fed inoculated silages had greater microbial N supply than those on the Control treatment (P<0.05). The acetate to propionate ratio was lower in ruminal fluid of wethers in LBLP treatment than LB and Control treatment (P<0.05) and ruminal pH tended to be greater in LB lambs than in LBLP and Control wethers (P<0.10). Finally, the inoculation with both bacteria combined enhanced the silage fermentation. The intakes of DM, OM, CP, NDF, and GE

  2. Soy protein hydrolysis with microbial protease to improve antioxidant and functional properties.

    PubMed

    de Oliveira, Cibele Freitas; Corrêa, Ana Paula Folmer; Coletto, Douglas; Daroit, Daniel Joner; Cladera-Olivera, Florencia; Brandelli, Adriano

    2015-05-01

    Soybean proteins are widely used as nutritional and functional food ingredients. This investigation evaluated through a 2(3) central composite design the effect of three variables (pH, temperature and enzyme/substrate (E/S) ratio) on the production of soy protein isolate (SPI) hydrolysates with a microbial protease. Soluble peptides, antioxidant activity, and foaming and emulsifying capabilities of the hydrolysates were analyzed. All variables, as well as their interactions, were significant for the soluble peptides content of SPI hydrolysates. Optimal conditions for obtaining soluble peptides were around 30-35 °C, pH 6.5-9.5, and E/S ratios of 1,650-6,300 U g(-1). SPI hydrolysates produced at 30-45 °C, pH 8.0-9.5, and E/S ratios of 4,000-8,000 U g(-1) showed higher capacity to scavenge the 2,2'-azino-bis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) radical. Models for soluble peptides and ABTS activity of hydrolysates were obtained. In the range studied, the variables had not significant influence on the ability of hydrolysates to scavenge the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. SPI hydrolysates also presented reducing power and ability to chelate iron. Hydrolysis temperature was significant for the Fe(2+)-chelating ability of hydrolysates. Temperature of hydrolysis was significant for the foaming capacity of hydrolysates, with higher values observed at 45 °C and 8,000 U g(-1). For emulsifying capacity, only E/S ratio presented a significant effect. Temperature and E/S ratio appeared to be more significant variables influencing the properties of the SPI hydrolysates. The results of this study indicate that specific hydrolysis conditions should be selected to obtain SPI hydrolysates with preferred characteristics.

  3. Cytochrome 572 is a conspicuous membrane protein with iron oxidation activity purified directly from a natural acidophilic microbial community.

    PubMed

    Jeans, Chris; Singer, Steven W; Chan, Clara S; Verberkmoes, Nathan C; Shah, Manesh; Hettich, Robert L; Banfield, Jillian F; Thelen, Michael P

    2008-05-01

    Recently, there has been intense interest in the role of electron transfer by microbial communities in biogeochemical systems. We examined the process of iron oxidation by microbial biofilms in one of the most extreme environments on earth, where the inhabited water is pH 0.5-1.2 and laden with toxic metals. To approach the mechanism of Fe(II) oxidation as a means of cellular energy acquisition, we isolated proteins from natural samples and found a conspicuous and novel cytochrome, Cyt(572), which is unlike any known cytochrome. Both the character of its covalently bound prosthetic heme group and protein sequence are unusual. Extraction of proteins directly from environmental biofilm samples followed by membrane fractionation, detergent solubilization and gel filtration chromatography resulted in the purification of an abundant yellow-red protein. The purified protein has a cytochrome c-type heme binding motif, CxxCH, but a unique spectral signature at 572 nm, and thus is called Cyt(572). It readily oxidizes Fe(2+) in the physiologically relevant acidic regime, from pH 0.95-3.4. Other physical characteristics are indicative of a membrane-bound multimeric protein. Circular dichroism spectroscopy indicates that the protein is largely beta-stranded, and 2D Blue-Native polyacrylamide gel electrophoresis and chemical crosslinking independently point to a multi-subunit structure for Cyt(572). By analyzing environmental genomic information from biofilms in several distinctly different mine locations, we found multiple genetic variants of Cyt(572). MS proteomics of extracts from these biofilms substantiated the prevalence of these variants in the ecosystem. Due to its abundance, cellular location and Fe(2+) oxidation activity at very low pH, we propose that Cyt(572) provides a critical function for fitness within the ecological niche of these acidophilic microbial communities.

  4. In situ expression of eukaryotic ice-binding proteins in microbial communities of Arctic and Antarctic sea ice

    PubMed Central

    Uhlig, Christiane; Kilpert, Fabian; Frickenhaus, Stephan; Kegel, Jessica U; Krell, Andreas; Mock, Thomas; Valentin, Klaus; Beszteri, Bánk

    2015-01-01

    Ice-binding proteins (IBPs) have been isolated from various sea-ice organisms. Their characterisation points to a crucial role in protecting the organisms in sub-zero environments. However, their in situ abundance and diversity in natural sea-ice microbial communities is largely unknown. In this study, we analysed the expression and phylogenetic diversity of eukaryotic IBP transcripts from microbial communities of Arctic and Antarctic sea ice. IBP transcripts were found in abundances similar to those of proteins involved in core cellular processes such as photosynthesis. Eighty-nine percent of the IBP transcripts grouped with known IBP sequences from diatoms, haptophytes and crustaceans, but the majority represented novel sequences not previously characterized in cultured organisms. The observed high eukaryotic IBP expression in natural eukaryotic sea ice communities underlines the essential role of IBPs for survival of many microorganisms in communities living under the extreme conditions of polar sea ice. PMID:25885562

  5. In situ expression of eukaryotic ice-binding proteins in microbial communities of Arctic and Antarctic sea ice.

    PubMed

    Uhlig, Christiane; Kilpert, Fabian; Frickenhaus, Stephan; Kegel, Jessica U; Krell, Andreas; Mock, Thomas; Valentin, Klaus; Beszteri, Bánk

    2015-11-01

    Ice-binding proteins (IBPs) have been isolated from various sea-ice organisms. Their characterisation points to a crucial role in protecting the organisms in sub-zero environments. However, their in situ abundance and diversity in natural sea-ice microbial communities is largely unknown. In this study, we analysed the expression and phylogenetic diversity of eukaryotic IBP transcripts from microbial communities of Arctic and Antarctic sea ice. IBP transcripts were found in abundances similar to those of proteins involved in core cellular processes such as photosynthesis. Eighty-nine percent of the IBP transcripts grouped with known IBP sequences from diatoms, haptophytes and crustaceans, but the majority represented novel sequences not previously characterized in cultured organisms. The observed high eukaryotic IBP expression in natural eukaryotic sea ice communities underlines the essential role of IBPs for survival of many microorganisms in communities living under the extreme conditions of polar sea ice.

  6. Complete genome sequence of Klebsiella pneumoniae J1, a protein-based microbial flocculant-producing bacterium.

    PubMed

    Pang, Changlong; Li, Ang; Cui, Di; Yang, Jixian; Ma, Fang; Guo, Haijuan

    2016-02-20

    Klebsiella pneumoniae J1 is a Gram-negative strain, which belongs to a protein-based microbial flocculant-producing bacterium. However, little genetic information is known about this species. Here we carried out a whole-genome sequence analysis of this strain and report the complete genome sequence of this organism and its genetic basis for carbohydrate metabolism, capsule biosynthesis and transport system. PMID:26806487

  7. Impact of high-pressure processing on microbial shelf-life and protein stability of refrigerated soymilk.

    PubMed

    Smith, K; Mendonca, A; Jung, S

    2009-12-01

    The effects of pressure (400, 500 and 600 MPa), dwell time (1 and 5 min) and temperature (25 and 75 degrees C) on microbial quality and protein stability of soymilk during 28 days of storage (4 degrees C) were evaluated under aerobic and anaerobic conditions. After processing and during storage, there were significant differences in total bacterial count (TBC), numbers of psychrotrophs (PSY) and Enterobacteriaceae (ENT), and protein stability between untreated (control) and pressurized samples (P < 0.05). Pressure applied at an initial temperature of 75 degrees C resulted in a greater suppression in growth of PSY compared to TBC. No ENT was detected in pressurized samples throughout the storage period tested. Dwell time had no significant effect on log reduction of TBC at 25 or 75 degrees C (P > 0.05). Pressure at 400 MPa (5 min), 500 and 600 MPa (1 and 5 min) produced 100% sub-lethal injury in surviving bacterial populations irrespective of temperature. After 28 days of refrigerated storage, both aerobic and anaerobic pressurized samples had better or similar stability as the control on day one of storage. Soymilk control samples were spoiled after 7 days whereas pressurization increased soymilk shelf-life by at least 2 weeks. Pressure (600 MPa) at 75 degrees C for 1 min not only significantly reduced initial microbial populations and increased the microbial shelf-life but also extended the protein stability of soymilk (P < 0.05).

  8. Efficiency and rumen responses in younger and older Holstein heifers limit-fed diets of differing energy density.

    PubMed

    Zanton, G I; Heinrichs, A J

    2016-04-01

    The objective of this study was to evaluate the effects of limit feeding diets of different predicted energy density on the efficiency of utilization of feed and nitrogen and rumen responses in younger and older Holstein heifers. Eight rumen-cannulated Holstein heifers (4 heifers beginning at 257 ± 7 d, hereafter "young," and 4 heifers beginning at 610 ± 16 d, hereafter "old") were limit-fed high [HED; 2.64 Mcal/kg of dry matter (DM), 15.31% crude protein (CP)] or low (LED; 2.42 Mcal/kg of DM, 14.15% CP) energy density diets according to a 4-period, split-plot Latin square design with 28-d periods. Diets were limit-fed to provide isonitrogenous and isoenergetic intake on a rumen empty body weight (BW) basis at a level predicted to support approximately 800 g/d of average daily gain. During the last 7d of each period, rumen contents were subsampled over a 24-h period, rumen contents were completely evacuated, and total collection of feces and urine was made over 4d. Intakes of DM and water were greater for heifers fed LED, although, by design, calculated intake of metabolizable energy did not differ between age groups or diets when expressed relative to rumen empty BW. Rumen pH was lower, ammonia (NH3-N) concentration tended to be higher, and volatile fatty acids (VFA) concentration was not different for HED compared with LED and was unaffected by age group. Rumen content mass was greater for heifers fed LED and for old heifers, so when expressing rumen fermentation responses corrected for this difference in pool size, NH3-N pool size was not different between diets and total moles of VFA in the rumen were greater for heifers fed LED, whereas these pool sizes were greater for old heifers. Total-tract digestibility of potentially digestible neutral detergent fiber (NDF) was greater in heifers fed LED and for young heifers, whereas the fractional rate of ruminal passage and digestion of NDF were both greater in heifers fed LED. Digestibility of N was greater for

  9. Efficiency and rumen responses in younger and older Holstein heifers limit-fed diets of differing energy density.

    PubMed

    Zanton, G I; Heinrichs, A J

    2016-04-01

    The objective of this study was to evaluate the effects of limit feeding diets of different predicted energy density on the efficiency of utilization of feed and nitrogen and rumen responses in younger and older Holstein heifers. Eight rumen-cannulated Holstein heifers (4 heifers beginning at 257 ± 7 d, hereafter "young," and 4 heifers beginning at 610 ± 16 d, hereafter "old") were limit-fed high [HED; 2.64 Mcal/kg of dry matter (DM), 15.31% crude protein (CP)] or low (LED; 2.42 Mcal/kg of DM, 14.15% CP) energy density diets according to a 4-period, split-plot Latin square design with 28-d periods. Diets were limit-fed to provide isonitrogenous and isoenergetic intake on a rumen empty body weight (BW) basis at a level predicted to support approximately 800 g/d of average daily gain. During the last 7d of each period, rumen contents were subsampled over a 24-h period, rumen contents were completely evacuated, and total collection of feces and urine was made over 4d. Intakes of DM and water were greater for heifers fed LED, although, by design, calculated intake of metabolizable energy did not differ between age groups or diets when expressed relative to rumen empty BW. Rumen pH was lower, ammonia (NH3-N) concentration tended to be higher, and volatile fatty acids (VFA) concentration was not different for HED compared with LED and was unaffected by age group. Rumen content mass was greater for heifers fed LED and for old heifers, so when expressing rumen fermentation responses corrected for this difference in pool size, NH3-N pool size was not different between diets and total moles of VFA in the rumen were greater for heifers fed LED, whereas these pool sizes were greater for old heifers. Total-tract digestibility of potentially digestible neutral detergent fiber (NDF) was greater in heifers fed LED and for young heifers, whereas the fractional rate of ruminal passage and digestion of NDF were both greater in heifers fed LED. Digestibility of N was greater for

  10. Medium Without Rumen Fluid for Nonselective Enumeration and Isolation of Rumen Bacteria

    PubMed Central

    Caldwell, Daniel R.; Bryant, Marvin P.

    1966-01-01

    Colony counts which approximated those in a habitat-simulating, rumen fluid-agar medium (RFM) were obtained in medium 10, a medium identical to the RFM except for the replacement of rumen fluid with 1.5 × 10-6m hemin, 0.2% Trypticase, 0.05% yeast extract, and a 6.6 × 10-2m volatile fatty acid mixture qualitatively and quantitatively similar to that in rumen fluid. Single deletion of Trypticase, yeast extract, or the volatile fatty acid mixture from medium 10 significantly reduced colony counts. Colony counts were also reduced when medium 10 was modified to contain higher concentrations of Trypticase or volatile fatty acids. Significant differences were found between colony counts obtained from diluted rumen contents of animals fed a cracked corn-urea diet, and the colony counts obtained from animals fed either a cracked corn-soyean oil meal or an alfalfa hay-grain diet. Qualitative differences were found between the predominant bacterial strains isolated from rumen contents of animals fed cracked corn diets and strains isolated from animals fed alfalfa hay-grain. Regardless of differences in the predominant flora associated with diet, medium 10 and the RFM supported growth of similar bacterial populations. The results show that medium 10 is suitable for enumeration and isolation of many predominant rumen bacteria. PMID:5970467

  11. Needles in the blue sea: sub-species specificity in targeted protein biomarker analyses within the vast oceanic microbial metaproteome.

    PubMed

    Saito, Mak A; Dorsk, Alexander; Post, Anton F; McIlvin, Matthew R; Rappé, Michael S; DiTullio, Giacomo R; Moran, Dawn M

    2015-10-01

    Proteomics has great potential for studies of marine microbial biogeochemistry, yet high microbial diversity in many locales presents us with unique challenges. We addressed this challenge with a targeted metaproteomics workflow for NtcA and P-II, two nitrogen regulatory proteins, and demonstrated its application for cyanobacterial taxa within microbial samples from the Central Pacific Ocean. Using METATRYP, an open-source Python toolkit, we examined the number of shared (redundant) tryptic peptides in representative marine microbes, with the number of tryptic peptides shared between different species typically being 1% or less. The related cyanobacteria Prochlorococcus and Synechococcus shared an average of 4.8 ± 1.9% of their tryptic peptides, while shared intraspecies peptides were higher, 13 ± 15% shared peptides between 12 Prochlorococcus genomes. An NtcA peptide was found to target multiple cyanobacteria species, whereas a P-II peptide showed specificity to the high-light Prochlorococcus ecotype. Distributions of NtcA and P-II in the Central Pacific Ocean were similar except at the Equator likely due to differential nitrogen stress responses between Prochlorococcus and Synechococcus. The number of unique tryptic peptides coded for within three combined oceanic microbial metagenomes was estimated to be ∼4 × 10(7) , 1000-fold larger than an individual microbial proteome and 27-fold larger than the human proteome, yet still 20 orders of magnitude lower than the peptide diversity possible in all protein space, implying that peptide mapping algorithms should be able to withstand the added level of complexity in metaproteomic samples.

  12. Improved Feed Protein Fractional Schemes for Formulating Rations With the Cornell Net Carbohydrate and Protein System

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adequate predictions of rumen-degradable protein (RDP) and rumen-undegradable protein (RUP) supplies are necessary to optimize performance while minimizing losses of excess nitrogen (N). The objectives of this study were to evaluate the original Cornell Net Carbohydrate Protein System (CNCPS) protei...

  13. Rumen Microorganisms Decrease Bioavailability of Inorganic Selenium Supplements.

    PubMed

    Galbraith, M L; Vorachek, W R; Estill, C T; Whanger, P D; Bobe, G; Davis, T Z; Hall, J A

    2016-06-01

    Despite the availability of selenium (Se)-enriched trace mineral supplements, we have observed low Se status in cattle and sheep offered traditional inorganic Se supplements. Reasons for this may include inadequate intake or low bioavailability of inorganic Se sources. The objective of this study was to determine whether rumen microorganisms (RMO) alter the bioavailability of Se sources commonly used in Se supplements. Rumen microorganisms were isolated from ewes (n = 4) and incubated ex vivo with no Se (control), with inorganic Na selenite or Na selenate, or with organic selenomethionine (SeMet). Total Se incorporated into RMO and the amount of elemental Se formed were determined under equivalent conditions. Incorporation of Se from Na selenite, Na selenate, or SeMet into RMO was measured as fold change compared with control (no added Se). Incorporation of Se into microbial mass was greater for SeMet (13.2-fold greater than no-Se control) compared with inorganic Se supplements (P = 0.02); no differences were observed between inorganic Na selenate (3.3-fold greater than no-Se control) and Na selenite (3.5-fold greater than no-Se control; P = 0.97). Formation of non-bioavailable, elemental Se was less for RMO incubated with SeMet compared with inorganic Se sources (P = 0.01); no differences were observed between Na selenate and Na selenite (P = 0.09). The clinical importance of these results is that the oral bioavailability of organic SeMet should be greater compared with inorganic Se sources because of greater RMO incorporation of Se and decreased formation of elemental Se by RMO.

  14. Rumen Microorganisms Decrease Bioavailability of Inorganic Selenium Supplements.

    PubMed

    Galbraith, M L; Vorachek, W R; Estill, C T; Whanger, P D; Bobe, G; Davis, T Z; Hall, J A

    2016-06-01

    Despite the availability of selenium (Se)-enriched trace mineral supplements, we have observed low Se status in cattle and sheep offered traditional inorganic Se supplements. Reasons for this may include inadequate intake or low bioavailability of inorganic Se sources. The objective of this study was to determine whether rumen microorganisms (RMO) alter the bioavailability of Se sources commonly used in Se supplements. Rumen microorganisms were isolated from ewes (n = 4) and incubated ex vivo with no Se (control), with inorganic Na selenite or Na selenate, or with organic selenomethionine (SeMet). Total Se incorporated into RMO and the amount of elemental Se formed were determined under equivalent conditions. Incorporation of Se from Na selenite, Na selenate, or SeMet into RMO was measured as fold change compared with control (no added Se). Incorporation of Se into microbial mass was greater for SeMet (13.2-fold greater than no-Se control) compared with inorganic Se supplements (P = 0.02); no differences were observed between inorganic Na selenate (3.3-fold greater than no-Se control) and Na selenite (3.5-fold greater than no-Se control; P = 0.97). Formation of non-bioavailable, elemental Se was less for RMO incubated with SeMet compared with inorganic Se sources (P = 0.01); no differences were observed between Na selenate and Na selenite (P = 0.09). The clinical importance of these results is that the oral bioavailability of organic SeMet should be greater compared with inorganic Se sources because of greater RMO incorporation of Se and decreased formation of elemental Se by RMO. PMID:26537117

  15. The complete genome sequence of the rumen methanogen Methanobrevibacter millerae SM9.

    PubMed

    Kelly, William J; Pacheco, Diana M; Li, Dong; Attwood, Graeme T; Altermann, Eric; Leahy, Sinead C

    2016-01-01

    Methanobrevibacter millerae SM9 was isolated from the rumen of a sheep maintained on a fresh forage diet, and its genome has been sequenced to provide information on the phylogenetic diversity of rumen methanogens with a view to developing technologies for methane mitigation. It is the first rumen isolate from the Methanobrevibacter gottschalkii clade to have its genome sequence completed. The 2.54 Mb SM9 chromosome has an average G + C content of 31.8 %, encodes 2269 protein-coding genes, and harbors a single prophage. The overall gene content is comparable to that of Methanobrevibacter ruminantium M1 and the type strain of M. millerae (ZA-10(T)) suggesting that the basic metabolism of these two hydrogenotrophic rumen methanogen species is similar. However, M. millerae has a larger complement of genes involved in methanogenesis including genes for methyl coenzyme M reductase II (mrtAGDB) which are not found in M1. Unusual features of the M. millerae genomes include the presence of a tannase gene which shows high sequence similarity with the tannase from Lactobacillus plantarum, and large non-ribosomal peptide synthase genes. The M. millerae sequences indicate that methane mitigation strategies based on the M. ruminantium M1 genome sequence are also likely to be applicable to members of the M. gottschalkii clade. PMID:27536339

  16. Isolation and characterization of novel lipases/esterases from a bovine rumen metagenome.

    PubMed

    Privé, Florence; Newbold, C Jamie; Kaderbhai, Naheed N; Girdwood, Susan G; Golyshina, Olga V; Golyshin, Peter N; Scollan, Nigel D; Huws, Sharon A

    2015-07-01

    Improving the health beneficial fatty acid content of meat and milk is a major challenge requiring an increased understanding of rumen lipid metabolism. In this study, we isolated and characterized rumen bacterial lipases/esterases using functional metagenomics. Metagenomic libraries were constructed from DNA extracted from strained rumen fluid (SRF), solid-attached bacteria (SAB) and liquid-associated rumen bacteria (LAB), ligated into a fosmid vector and subsequently transformed into an Escherichia coli host. Fosmid libraries consisted of 7,744; 8,448; and 7,680 clones with an average insert size of 30 to 35 kbp for SRF, SAB and LAB, respectively. Transformants were screened on spirit blue agar plates containing tributyrin for lipase/esterase activity. Five SAB and four LAB clones exhibited lipolytic activity, and no positive clones were found in the SRF library. Fosmids from positive clones were pyrosequenced and twelve putative lipase/esterase genes and two phospholipase genes retrieved. Although the derived proteins clustered into diverse esterase and lipase families, a degree of novelty was seen, with homology ranging from 40 to 78% following BlastP searches. Isolated lipases/esterases exhibited activity against mostly short- to medium-chain substrates across a range of temperatures and pH. The function of these novel enzymes recovered in ruminal metabolism needs further investigation, alongside their potential industrial uses. PMID:25575887

  17. Isolation and characterization of novel lipases/esterases from a bovine rumen metagenome.

    PubMed

    Privé, Florence; Newbold, C Jamie; Kaderbhai, Naheed N; Girdwood, Susan G; Golyshina, Olga V; Golyshin, Peter N; Scollan, Nigel D; Huws, Sharon A

    2015-07-01

    Improving the health beneficial fatty acid content of meat and milk is a major challenge requiring an increased understanding of rumen lipid metabolism. In this study, we isolated and characterized rumen bacterial lipases/esterases using functional metagenomics. Metagenomic libraries were constructed from DNA extracted from strained rumen fluid (SRF), solid-attached bacteria (SAB) and liquid-associated rumen bacteria (LAB), ligated into a fosmid vector and subsequently transformed into an Escherichia coli host. Fosmid libraries consisted of 7,744; 8,448; and 7,680 clones with an average insert size of 30 to 35 kbp for SRF, SAB and LAB, respectively. Transformants were screened on spirit blue agar plates containing tributyrin for lipase/esterase activity. Five SAB and four LAB clones exhibited lipolytic activity, and no positive clones were found in the SRF library. Fosmids from positive clones were pyrosequenced and twelve putative lipase/esterase genes and two phospholipase genes retrieved. Although the derived proteins clustered into diverse esterase and lipase families, a degree of novelty was seen, with homology ranging from 40 to 78% following BlastP searches. Isolated lipases/esterases exhibited activity against mostly short- to medium-chain substrates across a range of temperatures and pH. The function of these novel enzymes recovered in ruminal metabolism needs further investigation, alongside their potential industrial uses.

  18. Effects of intraruminal sodium chloride infusion on rumen and renal nitrogen and electrolyte dynamics in sheep.

    PubMed

    Godwin, I R; Williams, V J

    1986-09-01

    1. Sheep were given 800 g low-protein roughage/d at 2 h intervals and infused intraruminally with 0,500, 750, 1000, 1250, 1500 or 2000 mmol sodium chloride/d in 436 ml water. The digestibility of various food fractions and rumen ammonia, volatile fatty acids (VFA) and liquid turnover rate were measured, along with renal haemodynamics and the renal excretory patterns of nitrogen and electrolytes. Ad lib. food intake was determined during the infusion of 0 and 2000 mmol NaCl/d. 2. Infusion of NaCl up to 750 mmol/d had virtually no effect on the indices measured, except water intake and water excretion. Infusion of greater amounts caused a step-wise decrease in the digestibility of organic matter (OM) and N. Rumen liquid turnover rate was increased substantially and rumen NH3 and VFA concentrations were decreased. Ad lib. food intake was not different when either 0 or 2000 mmol NaCl/d were infused into the rumen. 3. The glomerular filtration rate and effective renal plasma flow (ERPF) were substantially increased after the infusion of 1250 mmol or more NaCl/d. Extracellular fluid volume was also increased. The renal excretion of urea and uric acid + allantoin (URAL) were decreased at the higher infusion rates but the fractional excretions of both these substances were enhanced. The excretion of sodium, chloride, calcium and magnesium were markedly increased with increasing salt infusion. 4. The results suggest that high NaCl inputs into the rumen increase the rumen turnover rate, which in turn decreases the digestibility of OM, particularly N. This causes lower rumen NH3 and VFA concentrations. Plasma urea and URAL concentrations are also decreased and this causes lower renal excretion of these substances despite a much higher fractional excretion resulting from the greatly enhanced urine flow rate. 5. When roughages low in N are given, NaCl intake should be kept below 20 mmol/kg body-weight per d to prevent a decline in the digestibility of the food and any

  19. Cytochrome 572 is a conspicuous membrane protein with iron oxidation activity purified directly from a natural acidophilic microbial community

    SciTech Connect

    Verberkmoes, Nathan C; Singer, Steven; Shah, Manesh B; Thelen, Michael P.; Hettich, Robert {Bob} L; Banfield, Jillian F.

    2008-01-01

    We have discovered and characterized a novel membrane cytochrome of an iron oxidizing microbial biofilm obtained from the surface of extremely acidic mine water. This protein was initially identified through proteogenomic analysis as one of many novel gene products of Leptospirillum group II, the dominant bacterium of this community (Ram et al, 2005, Science 308, 1915-20). Extraction of proteins directly from environmental biofilm samples followed by membrane fractionation, detergent solubilization and gel filtration chromatography resulted in the purification of an abundant yellow-red protein. Covalently bound to heme, the purified cytochrome has a unique spectral signature at 572 nm and is thus called Cyt572. It readily oxidizes Fe2+ even in the presence of Fe3+ over a pH range from 0.95 to 3.4. Independent experiments involving 2D blue-native polyacrylamide gel electrophoresis and chemical crosslinking establish a homotetrameric structure for Cyt572. Also, circular dichroism spectroscopy indicates that the protein is largely beta-stranded, consistent with an outer membrane location. Although no significant sequence homology to the full-length cytochrome is detected in protein databases, environmental DNA sequences from both Leptospirillum groups II and III reveal at least 17 strain variants of Cyt572. Due to its abundance, cellular location and Fe2+ oxidation activity, we propose Cyt572 is the iron oxidase of the Leptospirillum bacteria, providing a critical function for fitness within the ecological niche of this acidophilic microbial community.

  20. Autotrophic nitrogen assimilation and carbon capture for microbial protein production by a novel enrichment of hydrogen-oxidizing bacteria.

    PubMed

    Matassa, Silvio; Verstraete, Willy; Pikaar, Ilje; Boon, Nico

    2016-09-15

    Domestic used water treatment systems are currently predominantly based on conventional resource inefficient treatment processes. While resource recovery is gaining momentum it lacks high value end-products which can be efficiently marketed. Microbial protein production offers a valid and promising alternative by upgrading low value recovered resources into high quality feed and also food. In the present study, we evaluated the potential of hydrogen-oxidizing bacteria to upgrade ammonium and carbon dioxide under autotrophic growth conditions. The enrichment of a generic microbial community and the implementation of different culture conditions (sequenced batch resp. continuous reactor) revealed surprising features. At low selection pressure (i.e. under sequenced batch culture at high solid retention time), a very diverse microbiome with an important presence of predatory Bdellovibrio spp. was observed. The microbial culture which evolved under high rate selection pressure (i.e. dilution rate D = 0.1 h(-1)) under continuous reactor conditions was dominated by Sulfuricurvum spp. and a highly stable and efficient process in terms of N and C uptake, biomass yield and volumetric productivity was attained. Under continuous culture conditions the maximum yield obtained was 0.29 g cell dry weight per gram chemical oxygen demand equivalent of hydrogen, whereas the maximum volumetric loading rate peaked 0.41 g cell dry weight per litre per hour at a protein content of 71%. Finally, the microbial protein produced was of high nutritive quality in terms of essential amino acids content and can be a suitable substitute for conventional feed sources such as fishmeal or soybean meal. PMID:27262118

  1. Autotrophic nitrogen assimilation and carbon capture for microbial protein production by a novel enrichment of hydrogen-oxidizing bacteria.

    PubMed

    Matassa, Silvio; Verstraete, Willy; Pikaar, Ilje; Boon, Nico

    2016-09-15

    Domestic used water treatment systems are currently predominantly based on conventional resource inefficient treatment processes. While resource recovery is gaining momentum it lacks high value end-products which can be efficiently marketed. Microbial protein production offers a valid and promising alternative by upgrading low value recovered resources into high quality feed and also food. In the present study, we evaluated the potential of hydrogen-oxidizing bacteria to upgrade ammonium and carbon dioxide under autotrophic growth conditions. The enrichment of a generic microbial community and the implementation of different culture conditions (sequenced batch resp. continuous reactor) revealed surprising features. At low selection pressure (i.e. under sequenced batch culture at high solid retention time), a very diverse microbiome with an important presence of predatory Bdellovibrio spp. was observed. The microbial culture which evolved under high rate selection pressure (i.e. dilution rate D = 0.1 h(-1)) under continuous reactor conditions was dominated by Sulfuricurvum spp. and a highly stable and efficient process in terms of N and C uptake, biomass yield and volumetric productivity was attained. Under continuous culture conditions the maximum yield obtained was 0.29 g cell dry weight per gram chemical oxygen demand equivalent of hydrogen, whereas the maximum volumetric loading rate peaked 0.41 g cell dry weight per litre per hour at a protein content of 71%. Finally, the microbial protein produced was of high nutritive quality in terms of essential amino acids content and can be a suitable substitute for conventional feed sources such as fishmeal or soybean meal.

  2. Lyophilization of rumen fluid for use in culture media.

    PubMed Central

    Dehority, B A; Tirabasso, P A

    1989-01-01

    The supernatant from centrifugation at 1,000 x g of strained rumen fluid was lyophilized, and the residue and sublimate fractions were used to replace fresh rumen fluid in a complete roll tube medium for enumeration of total rumen bacteria. Most of the growth-supporting nutrients in fresh rumen fluid were found in the residue fraction. With one exception, no significant differences were found in total bacterial numbers either by roll tube or most-probable-number procedures when lyophilized rumen fluid residue was substituted for fresh rumen fluid. Lyophilized rumen fluid residue was stable for at least 5 months at room temperature. Rumen fluid supernatant from centrifugation at 1,000 x g had a mean density of 1.005 +/- 0.03 g/ml and contained 1.56% +/- 0.30% dry matter. On the basis of these values, 15.68 mg of lyophilized rumen fluid residue is equivalent to 1 ml of rumen fluid supernatant from centrifugation at 1,000 x g. PMID:2619310

  3. Taxonomic Assessment of Rumen Microbiota Using Total RNA and Targeted Amplicon Sequencing Approaches

    PubMed Central

    Li, Fuyong; Henderson, Gemma; Sun, Xu; Cox, Faith; Janssen, Peter H.; Guan, Le Luo

    2016-01-01

    Taxonomic characterization of active gastrointestinal microbiota is essential to detect shifts in microbial communities and functions under various conditions. This study aimed to identify and quantify potentially active rumen microbiota using total RNA sequencing and to compare the outcomes of this approach with the widely used targeted RNA/DNA amplicon sequencing technique. Total RNA isolated from rumen digesta samples from five beef steers was subjected to Illumina paired-end sequencing (RNA-seq), and bacterial and archaeal amplicons of partial 16S rRNA/rDNA were subjected to 454 pyrosequencing (RNA/DNA Amplicon-seq). Taxonomic assessments of the RNA-seq, RNA Amplicon-seq, and DNA Amplicon-seq datasets were performed using a pipeline developed in house. The detected major microbial phylotypes were common among the three datasets, with seven bacterial phyla, fifteen bacterial families, and five archaeal taxa commonly identified across all datasets. There were also unique microbial taxa detected in each dataset. Elusimicrobia and Verrucomicrobia phyla; Desulfovibrionaceae, Elusimicrobiaceae, and Sphaerochaetaceae families; and Methanobrevibacter woesei were only detected in the RNA-Seq and RNA Amplicon-seq datasets, whereas Streptococcaceae was only detected in the DNA Amplicon-seq dataset. In addition, the relative abundances of four bacterial phyla, eight bacterial families and one archaeal taxon were different among the three datasets. This is the first study to compare the outcomes of rumen microbiota profiling between RNA-seq and RNA/DNA Amplicon-seq datasets. Our results illustrate the differences between these methods in characterizing microbiota both qualitatively and quantitatively for the same sample, and so caution must be exercised when comparing data. PMID:27446027

  4. Taxonomic Assessment of Rumen Microbiota Using Total RNA and Targeted Amplicon Sequencing Approaches.

    PubMed

    Li, Fuyong; Henderson, Gemma; Sun, Xu; Cox, Faith; Janssen, Peter H; Guan, Le Luo

    2016-01-01

    Taxonomic characterization of active gastrointestinal microbiota is essential to detect shifts in microbial communities and functions under various conditions. This study aimed to identify and quantify potentially active rumen microbiota using total RNA sequencing and to compare the outcomes of this approach with the widely used targeted RNA/DNA amplicon sequencing technique. Total RNA isolated from rumen digesta samples from five beef steers was subjected to Illumina paired-end sequencing (RNA-seq), and bacterial and archaeal amplicons of partial 16S rRNA/rDNA were subjected to 454 pyrosequencing (RNA/DNA Amplicon-seq). Taxonomic assessments of the RNA-seq, RNA Amplicon-seq, and DNA Amplicon-seq datasets were performed using a pipeline developed in house. The detected major microbial phylotypes were common among the three datasets, with seven bacterial phyla, fifteen bacterial families, and five archaeal taxa commonly identified across all datasets. There were also unique microbial taxa detected in each dataset. Elusimicrobia and Verrucomicrobia phyla; Desulfovibrionaceae, Elusimicrobiaceae, and Sphaerochaetaceae families; and Methanobrevibacter woesei were only detected in the RNA-Seq and RNA Amplicon-seq datasets, whereas Streptococcaceae was only detected in the DNA Amplicon-seq dataset. In addition, the relative abundances of four bacterial phyla, eight bacterial families and one archaeal taxon were different among the three datasets. This is the first study to compare the outcomes of rumen microbiota profiling between RNA-seq and RNA/DNA Amplicon-seq datasets. Our results illustrate the differences between these methods in characterizing microbiota both qualitatively and quantitatively for the same sample, and so caution must be exercised when comparing data. PMID:27446027

  5. Studies on some characteristics of hydrogen production by cell-free extracts of rumen anaerobic bacteria.

    PubMed

    Joyner, A E; Winter, W T; Godbout, D M

    1977-03-01

    Hydrogen production was studied in the following rumen anaerobes: Bacteroides clostridiiformis, Butyrivibrio fibrisolvens, Enbacterium limosum, Fusobacterium necrophorum, Megasphaera elsdenii, Ruminococcus albus, and Ruminococcus flavefaciens. Clostridium pasteurianum and Escherichia coli were included for comparative purposes. Hydrogen production from dithionite, dithionite-reduced methyl viologen, pyruvate, and formate was determined. All species tested produced hydrogen from dithionite-reduce methyl viologen, but only C. pasteurianum, B. clostridiiformis, E. limosum, and M. elsdenii produced hydrogen from dithionite. All species except E. coli produced hydrogen from pyruvate, but activity was low or absent in extracts of E. limosum, F. necrophorum, R. albus, and R. flavefaciens unless methyl viologen was added. Hydrogen was produced from formate only by E. coli, B. clostridiiformis, E. limosum, F. necrophorum, and R. flavefaciens. Extracts were subjected to ultracentrifugation in an effort to determine the solubility of hydrogenase. The hydrogenase of all species except E. coli appeared to be soluble, although variable amounts of hydrogenase activity were detected in the pellet. Treatment of extracts of the rumen microbial species with DEAE-cellulose resulted in loss ofhydrogen production from pyruvate. Activity was restored by the addition of methyl viologen. It is concluded that hydrogen production in these rumen microorganisms is similar to that in the saccharolytic clostridia.

  6. Bacterial community dynamics in a rumen fluid bioreactor during in-vitro cultivation.

    PubMed

    Zapletalová, Martina; Kašparovská, Jitka; Křížová, Ludmila; Kašparovský, Tomáš; Šerý, Omar; Lochman, Jan

    2016-09-20

    To study the various processes in the rumen the in vitro techniques are widely used to realize more controlled and reproducible conditions compared to in vivo experiments. Mostly, only the parameters like pH changes, volatile fatty acids content or metabolite production are monitored. In this study we examine the bacterial community dynamics of rumen fluid in course of ten day cultivation realize under standard conditions described in the literature. Whereas the pH values, total VFA content and A/P ratio in bioreactor were consistent with natural conditions in the rumen, the mean redox-potential values of -251 and -243mV were much more negative. For culture-independent assessment of bacterial community composition, the Illumina MiSeq results indicated that the community contained 292 bacterial genera. In course of ten days cultivation a significant changes in the microbial community were measured when Bacteroidetes to Firmicutes ratio changed from 3.2 to 1.2 and phyla Proteobacteria and Actinobacteria represented by genus Bifidobacterium and Olsenella significantly increased. The main responsible factor of these changes seems to be very low redox potential in bioreactor together with accumulation of simple carbohydrates in milieu as a result of limited excretion of fermented feed and absence of nutrient absorbing mechanisms. PMID:27444706

  7. In vitro Screening of Essential Oil Active Compounds for Manipulation of Rumen Fermentation and Methane Mitigation.

    PubMed

    Joch, M; Cermak, L; Hakl, J; Hucko, B; Duskova, D; Marounek, M

    2016-07-01

    The objective of this study was to investigate the effects of 11 active compounds of essential oils (ACEO) on rumen fermentation characteristics and methane production. Two trials were conducted. In trial 1, ACEO (eugenol, carvacrol, citral, limonene, 1,4-cineole, p-cymene, linalool, bornyl acetate, α-pinene, and β-pinene) at a dose of 1,000 μL/L were incubated for 24 h in diluted rumen fluid with a 70:30 forage:concentrate substrate (16.2% crude protein; 36.6% neutral detergent fiber). Three fistulated Holstein cows were used as donors of rumen fluid. The reduction in methane production was observed with nine ACEO (up to 86% reduction) compared with the control (p<0.05). Among these, only limonene, 1,4-cineole, bornyl acetate, and α-pinene did not inhibit volatile fatty acid (VFA) production, and only bornyl acetate produced less methane per mol of VFA compared with the control (p<0.05). In a subsequent trial, the effects on rumen fermentation and methane production of two concentrations (500 and 2,000 μL/L) of bornyl acetate, the most promising ACEO from the first trial, were evaluated using the same in vitro incubation method that was used in the first trial. In trial 2, monensin was used as a positive control. Both doses of bornyl acetate decreased (p<0.05) methane production and did not inhibit VFA production. Positive effects of bornyl acetate on methane and VFA production were more pronounced than the effects of monensin. These results confirm the ability of bornyl acetate to decrease methane production, which may help to improve the efficiency of energy use in the rumen. PMID:26954157

  8. In vitro Screening of Essential Oil Active Compounds for Manipulation of Rumen Fermentation and Methane Mitigation

    PubMed Central

    Joch, M.; Cermak, L.; Hakl, J.; Hucko, B.; Duskova, D.; Marounek, M.

    2016-01-01

    The objective of this study was to investigate the effects of 11 active compounds of essential oils (ACEO) on rumen fermentation characteristics and methane production. Two trials were conducted. In trial 1, ACEO (eugenol, carvacrol, citral, limonene, 1,4-cineole, p-cymene, linalool, bornyl acetate, α-pinene, and β-pinene) at a dose of 1,000 μL/L were incubated for 24 h in diluted rumen fluid with a 70:30 forage:concentrate substrate (16.2% crude protein; 36.6% neutral detergent fiber). Three fistulated Holstein cows were used as donors of rumen fluid. The reduction in methane production was observed with nine ACEO (up to 86% reduction) compared with the control (p<0.05). Among these, only limonene, 1,4-cineole, bornyl acetate, and α-pinene did not inhibit volatile fatty acid (VFA) production, and only bornyl acetate produced less methane per mol of VFA compared with the control (p<0.05). In a subsequent trial, the effects on rumen fermentation and methane production of two concentrations (500 and 2,000 μL/L) of bornyl acetate, the most promising ACEO from the first trial, were evaluated using the same in vitro incubation method that was used in the first trial. In trial 2, monensin was used as a positive control. Both doses of bornyl acetate decreased (p<0.05) methane production and did not inhibit VFA production. Positive effects of bornyl acetate on methane and VFA production were more pronounced than the effects of monensin. These results confirm the ability of bornyl acetate to decrease methane production, which may help to improve the efficiency of energy use in the rumen. PMID:26954157

  9. Nonfeed application of rendered animal proteins for microbial production of eicosapentaenoic acid by the fungus Pythium irregulare.

    PubMed

    Liang, Yi; Garcia, Rafael A; Piazza, George J; Wen, Zhiyou

    2011-11-23

    Rendered animal proteins are well suited for animal nutrition applications, but the market is maturing, and there is a need to develop new uses for these products. The objective of this study is to explore the possibility of using animal