Genetics of the proteolytic system of lactic acid bacteria

NARCIS (Netherlands)

Kok, Jan

1990-01-01

The proteolytic system of lactic acid bacteria is of eminent importance for the rapid growth of these organisms in protein-rich media. The combined action of proteinases and peptidases provides the cell with small peptides and essential amino acids. The amino acids and peptides thus liberated have

Effect of feeding palm oil by-products based diets on total bacteria, cellulolytic bacteria and methanogenic archaea in the rumen of goats.

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Abubakr, Abdelrahim; Alimon, Abdul Razak; Yaakub, Halimatun; Abdullah, Norhani; Ivan, Michael

2014-01-01

Rumen microorganisms are responsible for digestion and utilization of dietary feeds by host ruminants. Unconventional feed resources could be used as alternatives in tropical areas where feed resources are insufficient in terms of quality and quantity. The objective of the present experiment was to evaluate the effect of diets based on palm oil (PO), decanter cake (DC) or palm kernel cake (PKC) on rumen total bacteria, selected cellulolytic bacteria, and methanogenic archaea. Four diets: control diet (CD), decanter cake diet (DCD), palm kernel cake diet (PKCD) and CD plus 5% PO diet (CPOD) were fed to rumen cannulated goats and rumen samples were collected at the start of the experimental diets (day 0) and on days 4, 6, 8, 12, 18, 24 and 30 post dietary treatments. Feeding DCD and PKCD resulted in significantly higher (Pgoats fed PKCD and CPOD and the trend showed a severe reduction on days 4 and 6 post experimental diets. In conclusion, results indicated that feeding DCD and PKC increased the populations of cellulolytic bacteria and decreased the density of methanogenic archaea in the rumen of goats.

Pathogenic Assay of Probiotic Bacteria Producing Proteolytic Enzymes as Bioremediation Bacteria Against Vannamei Shrimp Larvae (Litopenaeus vannamei)

OpenAIRE

Wilis Ari Setyati; Muhammad Zainuddin; Person Pesona Renta

2017-01-01

Application of bacteria in bioremediation of shrimp culture ponds is one of the methods used to clean internal pollutants. This study aimed to evaluate the pathogenicity of extracellular proteolytic enzyme produced by the probiotic bacteria as bioremediation bacteria on vannamei shrimp larvae culture. There were five probiotic bacteria, which were successfully isolated from the sediments served as substrate in mangrove area. The isolated bacteria were coded in number as 13, 19, 30, 33, and 36...

Isolation of previously uncultured rumen bacteria by dilution to extinction using a new liquid culture medium.

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Kenters, Nikki; Henderson, Gemma; Jeyanathan, Jeyamalar; Kittelmann, Sandra; Janssen, Peter H

2011-01-01

A new anaerobic medium that mimics the salts composition of rumen fluid was used in conjunction with a dilution method of liquid culture to isolate fermentative bacteria from the rumen of a grass-fed sheep. The aim was to inoculate a large number of culture tubes each with a mean of 97% sequence identity to genes of uncultured bacteria detected in various gastrointestinal environments. This strategy has therefore allowed us to cultivate many novel rumen bacteria, opening the way to overcoming the lack of cultures of many of the groups detected using cultivation-independent methods. Copyright © 2010 Elsevier B.V. All rights reserved.

The effect of rumen content transfer on rate of bacteria and protozoa growth

International Nuclear Information System (INIS)

Suharyono; M Winugroho; Y Widiati; S Marijati

1998-01-01

The aims the experiment wants to know the benefit of rate of microbial protein in rumen content and to complete the information that isolates is useful for ruminant animals feed. The result indicated that buffaloes from East Nusa Tenggara is the best when they are used as donor rumen transfer making isolate. When rumen of ongole cattle generation was mixed in rumen content of buffaloes from East Nusa Tenggara and incubated 48 h, the rate of bacteria cell growth is better than rate of protozoa cell growth comparing to the other animals. The values are 30.99 mg/h/100 ml and 24.92 mg.h/100 ml respectively. The results of isolate selection in 48 h incubation indicated that treatment F is the best. The results rates of bacteria cell growth and rate of protozoa's cell growth are 26.96 mg/h/100 ml and 2.53 mg/h/100 respectively. The result of in vitro study indicated that pH and ammonia concentration support the rate of bacteria cell growth and do not cause the toxicity of microbes and animal . The rate of bacteria cell growth on D treatment is significant to A,B, and C treatments. The values are 21.44 mg/h/100 ml. 7.99; 13.13; and 13.38 mg/h/100 ml respectively. The result rates of protozoa's cell growth tends lower than rates of bacteria cell. The overall conclusion is a lower or a higher rate of microorganism cell growth depends on the environment condition. (author)

Maintenance of Laboratory Strains of Obligately Anaerobic Rumen Bacteria †

Science.gov (United States)

Teather, Ronald M.

1982-01-01

Cultures of rumen bacteria can be stored at −20°C for at least 2 years in a liquid medium containing 20% glycerol. Thawing, sampling, and refreezing do not significantly affect viability. PMID:7125660

Accumulation of reserve carbohydrate by rumen protozoa and bacteria in competition for glucose.

Science.gov (United States)

Denton, Bethany L; Diese, Leanne E; Firkins, Jeffrey L; Hackmann, Timothy J

2015-03-01

The aim of this study was to determine if rumen protozoa could form large amounts of reserve carbohydrate compared to the amounts formed by bacteria when competing for glucose in batch cultures. We separated large protozoa and small bacteria from rumen fluid by filtration and centrifugation, recombined equal protein masses of each group into one mixture, and subsequently harvested (reseparated) these groups at intervals after glucose dosing. This method allowed us to monitor reserve carbohydrate accumulation of protozoa and bacteria individually. When mixtures were dosed with a moderate concentration of glucose (4.62 or 5 mM) (n = 2 each), protozoa accumulated large amounts of reserve carbohydrate; 58.7% (standard error of the mean [SEM], 2.2%) glucose carbon was recovered from protozoal reserve carbohydrate at time of peak reserve carbohydrate concentrations. Only 1.7% (SEM, 2.2%) was recovered in bacterial reserve carbohydrate, which was less than that for protozoa (P protozoa can sequester sugar away from bacteria by accumulating reserve carbohydrate, giving protozoa a competitive advantage and stabilizing fermentation in the rumen. Similar experiments could be used to investigate the importance of starch sequestration. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Isolation and Identification of Sodium Fluoroacetate Degrading Bacteria from Caprine Rumen in Brazil

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Expedito K. A. Camboim

2012-01-01

Full Text Available The objective of this paper was to report the isolation of two fluoroacetate degrading bacteria from the rumen of goats. The animals were adult goats, males, crossbred, with rumen fistula, fed with hay, and native pasture. The rumen fluid was obtained through the rumen fistula and immediately was inoculated 100 μL in mineral medium added with 20 mmol L−1 sodium fluoroacetate (SF, incubated at 39°C in an orbital shaker. Pseudomonas fluorescens (strain DSM 8341 was used as positive control for fluoroacetate dehalogenase activity. Two isolates were identified by 16S rRNA gene sequencing as Pigmentiphaga kullae (ECPB08 and Ancylobacter dichloromethanicus (ECPB09. These bacteria degraded sodium fluoroacetate, releasing 20 mmol L−1 of fluoride ion after 32 hours of incubation in Brunner medium containing 20 mmol L−1 of SF. There are no previous reports of fluoroacetate dehalogenase activity for P. kullae and A. dichloromethanicus. Control measures to prevent plant intoxication, including use of fences, herbicides, or other methods of eliminating poisonous plants, have been unsuccessful to avoid poisoning by fluoroacetate containing plants in Brazil. In this way, P. kullae and A. dichloromethanicus may be used to colonize the rumen of susceptible animals to avoid intoxication by fluoroacetate containing plants.

Labelled Thioamino Acids to Indicate the Synthetic Activity of the Rumen Bacteria in In-Vitro Experiments

Energy Technology Data Exchange (ETDEWEB)

Panic, B.; Jovanovic, M.; Cuperlovic, M.; Djordjevic, D. [Institute for the Application of Nuclear Energy in Agriculture, Veterinary Medicine and Forestry, Belgrade, Yugoslavia (Serbia)

1968-07-01

The synthetic activity of rumen bacteria has been studied in vitro through the investigation of cystine and methionine concentration and their specific activity. {sup 35}S-sulphate has been used as a radioactive tracer. Two diets, different in the level of nutrients - energy and protein - were added to the artificial tumen. The incubation with bacteria from the rumen content of the cows, fed for four weeks with the same diet, lasted 19 h. The diet with the higher level of protein and energy increased the cystine content (per 100 mg of N{sub 2}) by 23.3% and the methionine content by 39.4%. The concentration of radioactive cystine was increased at the same percentage rate by 25%, but radioactive methionine was much lower and increased only 6.4%. The difference between the specific activities of the investigated amino acids can be explained by the different catabolism rate and utilization of dietary cystine, and methionine by the rumen bacterial flora. Since the dietary methionine is catabolized slowly, it can, especially by the use of the diets with a high protein level, significantly decrease the specific activity of the radioactive methionine synthesized by rumen bacteria. Therefore, the incorporation of {sup 35}S into the cystine represents a more reliable indicator of the synthetic activity of the rumen bacteria. (author)

Roles of Dietary Cobalt and Administration of Mixed Rumen Bacteria in Regulating Hematological Parameters of Pre-weaning Twin Lambs

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T. Adelina

2013-08-01

Full Text Available Cobalt (Co is required by rumen microorganism for vitamin B12 synthesis. Vitamin B12 is an important cofactor for methionine synthesis and gluconeogenesis. In young ruminants up to 6–8 wk old, the rumen has not been completely developed and rumen microorganisms are not ready to supply vitamin B12. The aim of this research was to determine the potency of mixed rumen bacteria and dietary supplementation of Co and its effect on plasma glucose, blood minerals (Co, Fe, and Zn concentrations, and hematology of pre-weaning twin lambs. Twelve one month-old local twin lambs were assigned to 4 groups in a randomized complete block design. Lambs were fed cow milk at 10% body weight, adjusted weekly for 80 d. Mixed rumen bacteria were offered at 15 mL/d (8.295x1010 cfu. Dietary treatments were: 1 basal diet (Control, 2 basal diet + 1 mg/kg DM cyanocobalamin (VitB12 and 3 basal diet + 1 mg/kg DM of Co + administration of 15 mL mixed rumen bacteria (CoBac. There were no treatment effects on neither plasma glucose and blood mineral concentrations nor hematological profiles. This study demonstrated that pre-weaning twin lambs are not responsive to supplementation of Co and administration of mixed rumen bacteria.

Degradation of spent craft brewer's yeast by caprine rumen hyper ammonia-producing bacteria.

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Harlow, B E; Bryant, R W; Cohen, S D; O'Connell, S P; Flythe, M D

2016-10-01

Spent yeast from craft beers often includes more hops (Humulus lupulus L.) secondary metabolites than traditional recipes. These compounds include α- and β- acids, which are antimicrobial to the rumen hyper ammonia-producing bacteria (HAB) that are major contributors to amino acid degradation. The objective was to determine if the hops acids in spent craft brewer's yeast (CY; ~ 3·5 mg g(-1) hops acids) would protect it from degradation by caprine rumen bacteria and HAB when compared to a baker's yeast (BY; no hops acids). Cell suspensions were prepared by harvesting rumen fluid from fistulated goats, straining and differential centrifugation. The cells were re-suspended in media with BY or CY. After 24 h (39°C), HAB were enumerated and ammonia was measured. Fewer HAB and less ammonia was produced from CY than from BY. Pure culture experiments were conducted with Peptostreptococcus anaerobiusBG1 (caprine HAB). Ammonia production by BG1 from BY was greater than from CY. Ammonia production was greater when exogenous amino acids were included, but similar inhibition was observed in CY treatments. These results indicate that rumen micro-organisms deaminated the amino acids in CY to a lesser degree than BY. 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.) than traditional recipes. These compounds include α- and β- acids, which are antimicrobial to the rumen hyper ammonia-producing bacteria (HAB) that are major contributors to amino acid degradation. This study demonstrated that hops acids in spent craft brewer's yeast protected protein from destruction by HABin vitro. These results suggest that the spent yeast from craft breweries, a source of beneficial hops secondary metabolites, could have value as rumen-protected protein. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

Effects of Supplementation of Eucalyptus ( Leaf Meal on Feed Intake and Rumen Fermentation Efficiency in Swamp Buffaloes

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N. T. Thao

2015-07-01

Full Text Available 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.

Comparing the responses of rumen ciliate protozoa and bacteria to excess carbohydrate.

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Teixeira, César R V; Lana, Rogério de Paula; Tao, Junyi; Hackmann, Timothy J

2017-06-01

When given excess carbohydrate, certain microbial species respond by storing energy (synthesizing reserve carbohydrate), but other species respond by dissipating the energy as heat (spilling energy). To determine the importance of these responses in the rumen microbial community, this study quantified the responses of mixed ciliate protozoa vs bacteria to glucose. We hypothesized that ciliates would direct more glucose to synthesis of reserve carbohydrate (and less to energy spilling) than would bacteria. Ciliates and bacteria were isolated from rumen fluid using filtration and centrifugation, resuspended in nitrogen-free buffer to limit growth, and dosed with 5 mM glucose. Compared with bacteria, ciliates consumed glucose >3-fold faster and synthesized reserve carbohydrate 4-fold faster. They incorporated 53% of glucose carbon into reserve carbohydrate-nearly double the value (27%) for bacteria. Energy spilling was not detected for ciliates, as all heat production (104%) was accounted by synthesis of reserve carbohydrate and endogenous metabolism. For bacteria, reserve carbohydrate and endogenous metabolism accounted for only 68% of heat production, and spilling was detected within 11 min of dosing glucose. These results suggest that ciliates alter the course of ruminal carbohydrate metabolism by outcompeting bacteria for excess carbohydrate, maximizing reserve carbohydrate synthesis, and minimizing energy spilling. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Effects of alkyl polyglycoside, a nonionic surfactant, and forage-to-concentrate ratio on rumen fermentation, amino acid composition of rumen content, bacteria and plasma in goats.

Science.gov (United States)

Zeng, Bo; Tan, Zhiliang; Tang, Shaoxun; Han, Xuefeng; Tan, Chuanyan; Zhong, Rongzhen; Hea, Zhixiong; Arigbede, Oluwasanmi Moses

2011-06-01

In the present study, the effects of different forage-to-concentrate ratios (F:C) and an alkyl polyglycoside (APG) supplementation on parameters of rumen and blood metabolism were investigated in goats. A 2 x 2 factorial experiment was arranged within a 4 x 4 Latin square design (four 22-day periods), using four wether goats equipped with permanent ruminal cannulas. The experimental diets included two F:C levels (40:60 vs. 60:40), and two APG supplementation levels (None or 13 ml APG daily per animal). Rumen contents and blood samples were collected at the end of each period. Dietary F:C alteration affected plasma urea and influenced the proportions of leucine, histidine, arginine, glycine, proline, alanine, valine, phenylalanine, cysteine and tyrosine in rumen content, and the proportions of methionine, threonine and proline in solid-associated bacteria (SAB) significantly. Dietary APG decreased the proportions of valine and phenylalanine in rumen content, and the histidine content of liquid-associated bacteria. The interaction between dietary F:C and APG was significant for the proportions of glycine and alanine in rumen content, and the proportions of lysine and threonine in SAB. The proportion of lysine was greater, but the proportion of threonine was less in SAB for goats fed high F:C diet without APG supplementation. The proportions of plasma free amino acids and glucose concentration were not affected by experimental treatments. These results indicated that dietary APG addition affected the amino acid composition of the rumen content and ruminal bacteria, but this depended on the dietary F:C ratio. It is necessary to validate the effectiveness of dietary APG supplementation in further studies with more animals.

Potential of Selected Rumen Bacteria for Cellulose and Hemicellulose Degradation

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Maša Zorec

2014-01-01

Full Text Available Herbivorous animals harbour potent cellulolytic and hemicellulolytic microorganisms that supply the host with nutrients acquired from degradation of ingested plant material. In addition to protozoa and fungi, rumen bacteria contribute a considerable part in the breakdown of recalcitrant (hemicellulosic biomass. The present review is focused on the enzymatic systems of three representative fibrolytic rumen bacteria, namely Ruminococcus flavefaciens, Prevotella bryantii and Pseudobutyrivibrio xylanivorans. R. flavefaciens is known for one of the most elaborated cellulosome architectures and might represent a promising candidate for the construction of designer cellulosomes. On the other hand, Prevotella bryantii and Pseudobutyrivibrio xylanivorans produce multiple free, but highly efficient xylanases. In addition, P. xylanivorans was also shown to have some probiotic traits, which makes it a promising candidate not only for biogas production, but also as an animal feed supplement. Genomic and proteomic analyses of cellulolytic and hemicellulolytic bacterial species aim to identify novel enzymes, which can then be cloned and expressed in adequate hosts to construct highly active recombinant hydrolytic microorganisms applicable for different biotechnological tasks.

Proteolytic activity and cooperative hemolytic effect of dermatophytes with different species of bacteria

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Pakshir, K; Mohamadi, T; Khodadadi, H; Motamedifar, M; Zomorodian, K; Alipour, S; Motamedi, M

2016-01-01

Background and Purpose: Globally, dermatophytes are the most common filamentous group of fungi causing cutaneous mycoses. Dermatophytes were shown to secrete a multitude of enzymes that play a role in their pathogenesis. There is limited data on co-hemolytic (CAMP-like) effect of different bacterial species on dermatophyte species. In this study, we sought to the evaluate exoenzyme activity and co-hemolytic effect of four bacteria on clinical dermatophytes isolated from patients in Shiraz, Iran. Materials and Methods: A total of 84 clinical dermatophyte species were isolated from patients suffering dermatophytosis and identified by conventional methods. Hemolytic activity was evaluated with Columbia 5% sheep blood agar. Proteolytic activity was determined by plate clearance assay method, using gelatin 8% agar. CAMP-like factor was evaluated with four bacteria, namely, S. areus, S. saprophyticus, S. pyogenes, and S. agalactiae. Fisher's exact test was run for statistical analysis. Results: T. mentagrophytes was the most predominant agent (27 [32.1%]) followed by T. verrucosum(20 [23.8%]), T. tonsurans (10 [11.9%]), Microsporum canis (7 [8.3%]), T. rubrum (6 [7.1%]), E. floccosum (6 [7.1%]), M. gypseum (5 [6%]), and T. violaceum (3[3.6%]). The most common clinical area of dermatophytosis was the skin. All the isolates expressed the zone of incomplete alpha hemolysis. All the isolates had CAMP- positive reaction with S. aureus and the other bacteria were CAMP-negative. All the isolates expressed proteolytic activity and no significant differences were noted among diverse genera of dermatophytes and severities of proteolytic activity. Conclusion: This study indicated that hemolysin and proteolytic enzymes potentially play a role in dermatophyte pathogenesis and S. aureus could be considered as a main bacterium for creation of co-hemolytic effect in association with dermatophyte species. PMID:28959790

Comparison of rumen bacteria distribution in original rumen digesta, rumen liquid and solid fractions in lactating Holstein cows

OpenAIRE

Ji, Shoukun; Zhang, Hongtao; Yan, Hui; Azarfar, Arash; Shi, Haitao; Alugongo, Gibson; Li, Shengli; Cao, Zhijun; Wang, Yajing

2017-01-01

Background Original rumen digesta, rumen liquid and solid fractions have been frequently used to assess the rumen bacterial community. However, bacterial profiles in rumen original digesta, liquid and solid fractions vary from each other and need to be better established. Methods To compare bacterial profiles in each fraction, samples of rumen digesta from six cows fed either a high fiber diet (HFD) or a high energy diet (HED) were collected via rumen fistulas. Rumen digesta was then squeezed...

Rumen bacteria: interaction with particulate dietary components and response to dietary variation.

Science.gov (United States)

Cheng, K J; Akin, D E; Costerton, J W

1977-02-01

The bovine rumen resembles many other ecosystems in that its component bacterial cells are universally surrounded and protected by extracellular structures. The most common form of these structures is a fibrous carbohydrate slime that extends away from the cell and may mediate the attachment of the bacterium to a surface. This attachment is relatively specific and it may occur at the surface of the rumen epithelium or on the cell walls of a specific tissue within the plant-derived food of the animal. The production of the extracellular slime is under nutritional control and slime may be overproduced when soluble carbohydrates are available in high concentration. This overproduction results in cell-cell adhesion among the rumen bacteria with the eventual formation of slime-enclosed microcolonies and, in extreme cases, the generation of sufficient viscosity to cause feedlot bloat.

Alteration of Rumen Bacteria and Protozoa Through Grazing Regime as a Tool to Enhance the Bioactive Fatty Acid Content of Bovine Milk.

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Bainbridge, Melissa L; Saldinger, Laurel K; Barlow, John W; Alvez, Juan P; Roman, Joe; Kraft, Jana

2018-01-01

Rumen microorganisms are the origin of many bioactive fatty acids (FA) found in ruminant-derived food products. Differences in plant leaf anatomy and chemical composition between cool- and warm-season pastures may alter rumen microorganisms, potentially enhancing the quantity/profile of bioactive FA available for incorporation into milk. The objective of this study was to identify rumen bacteria and protozoa and their cellular FA when cows grazed a warm-season annual, pearl millet (PM), in comparison to a diverse cool-season pasture (CSP). Individual rumen digesta samples were obtained from five Holstein cows in a repeated measures design with 28-day periods. The treatment sequence was PM, CSP, then PM. Microbial DNA was extracted from rumen digesta and sequence reads were produced with Illumina MiSeq. Fatty acids (FA) were identified in rumen bacteria and protozoa using gas-liquid chromatography/mass spectroscopy. Microbial communities shifted in response to grazing regime. Bacteria of the phylum Bacteroidetes were more abundant during PM than CSP ( P rumenic acid, and α-linolenic acid in milk. In conclusion, grazing regime can potentially be used to alter microbial communities shifting the FA profile of microbial cells, and subsequently, alter the milk FA profile.

Dose and time response of ruminally infused algae on rumen fermentation characteristics, biohydrogenation and Butyrivibrio group bacteria in goats.

Science.gov (United States)

Zhu, Honglong; Fievez, Veerle; Mao, Shengyong; He, Wenbo; Zhu, Weiyun

2016-01-01

Micro-algae could inhibit the complete rumen BH of dietary 18-carbon unsaturated fatty acid (UFAs). This study aimed to examine dose and time responses of algae supplementation on rumen fermentation, biohydrogenation and Butyrivibrio group bacteria in goats. Six goats were used in a repeated 3 × 3 Latin square design, and offered a fixed diet. Algae were infused through rumen cannule with 0 (Control), 6.1 (L-Alg), or 18.3 g (H-Alg) per day. Rumen contents were sampled on d 0, 3, 7, 14 and 20. H-Alg reduced total volatile fatty acid concentration and acetate molar proportion (P Algae induced a dose-dependent decrease in 18:0 and increased trans-18:1 in the ruminal content (P Algae had no effect on the abundances of Butyrivibrio spp. and Butyrivibrio proteoclasticus (P > 0.10), while H-Alg reduced the total bacteria abundance (P algae were related to the supplementation level, but there was no evidence of shift in ruminal biohydrogenation pathways towards t10-18:1. L-Alg mainly induced a transient effect on rumen biohydrogenation of 18-carbon UFAs, while H-Alg showed an acute inhibition and these effects were not associated with the known hydrogenating bacteria.

Horizontal gene transfer from Bacteria to rumen Ciliates indicates adaptation to their anaerobic, carbohydrates-rich environment

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Takenaka Akio

2006-02-01

Full Text Available Abstract Background The horizontal transfer of expressed genes from Bacteria into Ciliates which live in close contact with each other in the rumen (the foregut of ruminants was studied using ciliate Expressed Sequence Tags (ESTs. More than 4000 ESTs were sequenced from representatives of the two major groups of rumen Cilates: the order Entodiniomorphida (Entodinium simplex, Entodinium caudatum, Eudiplodinium maggii, Metadinium medium, Diploplastron affine, Polyplastron multivesiculatum and Epidinium ecaudatum and the order Vestibuliferida, previously called Holotricha (Isotricha prostoma, Isotricha intestinalis and Dasytricha ruminantium. Results A comparison of the sequences with the completely sequenced genomes of Eukaryotes and Prokaryotes, followed by large-scale construction and analysis of phylogenies, identified 148 ciliate genes that specifically cluster with genes from the Bacteria and Archaea. The phylogenetic clustering with bacterial genes, coupled with the absence of close relatives of these genes in the Ciliate Tetrahymena thermophila, indicates that they have been acquired via Horizontal Gene Transfer (HGT after the colonization of the gut by the rumen Ciliates. Conclusion Among the HGT candidates, we found an over-representation (>75% of genes involved in metabolism, specifically in the catabolism of complex carbohydrates, a rich food source in the rumen. We propose that the acquisition of these genes has greatly facilitated the Ciliates' colonization of the rumen providing evidence for the role of HGT in the adaptation to new niches.

Horizontal gene transfer from Bacteria to rumen Ciliates indicates adaptation to their anaerobic, carbohydrates-rich environment.

Science.gov (United States)

Ricard, Guénola; McEwan, Neil R; Dutilh, Bas E; Jouany, Jean-Pierre; Macheboeuf, Didier; Mitsumori, Makoto; McIntosh, Freda M; Michalowski, Tadeusz; Nagamine, Takafumi; Nelson, Nancy; Newbold, Charles J; Nsabimana, Eli; Takenaka, Akio; Thomas, Nadine A; Ushida, Kazunari; Hackstein, Johannes H P; Huynen, Martijn A

2006-02-10

The horizontal transfer of expressed genes from Bacteria into Ciliates which live in close contact with each other in the rumen (the foregut of ruminants) was studied using ciliate Expressed Sequence Tags (ESTs). More than 4000 ESTs were sequenced from representatives of the two major groups of rumen Cilates: the order Entodiniomorphida (Entodinium simplex, Entodinium caudatum, Eudiplodinium maggii, Metadinium medium, Diploplastron affine, Polyplastron multivesiculatum and Epidinium ecaudatum) and the order Vestibuliferida, previously called Holotricha (Isotricha prostoma, Isotricha intestinalis and Dasytricha ruminantium). A comparison of the sequences with the completely sequenced genomes of Eukaryotes and Prokaryotes, followed by large-scale construction and analysis of phylogenies, identified 148 ciliate genes that specifically cluster with genes from the Bacteria and Archaea. The phylogenetic clustering with bacterial genes, coupled with the absence of close relatives of these genes in the Ciliate Tetrahymena thermophila, indicates that they have been acquired via Horizontal Gene Transfer (HGT) after the colonization of the gut by the rumen Ciliates. Among the HGT candidates, we found an over-representation (>75%) of genes involved in metabolism, specifically in the catabolism of complex carbohydrates, a rich food source in the rumen. We propose that the acquisition of these genes has greatly facilitated the Ciliates' colonization of the rumen providing evidence for the role of HGT in the adaptation to new niches.

Horizontal gene transfer from Bacteria to rumen Ciliates indicates adaptation to their anaerobic, carbohydrates-rich environment

OpenAIRE

Ricard, Guénola; McEwan, Neil R; Dutilh, Bas E; Jouany, Jean-Pierre; Macheboeuf, Didier; Mitsumori, Makoto; McIntosh, Freda M; Michalowski, Tadeusz; Nagamine, Takafumi; Nelson, Nancy; Newbold, Charles J; Nsabimana, Eli; Takenaka, Akio; Thomas, Nadine A; Ushida, Kazunari

2006-01-01

Abstract Background The horizontal transfer of expressed genes from Bacteria into Ciliates which live in close contact with each other in the rumen (the foregut of ruminants) was studied using ciliate Expressed Sequence Tags (ESTs). More than 4000 ESTs were sequenced from representatives of the two major groups of rumen Cilates: the order Entodiniomorphida (Entodinium simplex, Entodinium caudatum, Eudiplodinium maggii, Metadinium medium, Diploplastron affine, Polyplastron multivesiculatum and...

Rumen

DEFF Research Database (Denmark)

Mackie, Roderick I.; McSweeney, Christopher S.; Aminov, Rustam

2013-01-01

The rumen is a large pregastric fermentation compartment (foregut), which maintains a diverse but concentrated population of anaerobic bacteria, protozoa and fungi that are responsible for a variety of degradative and fermentative reactions. During this process biodegradable organic matter, mainly......). The concept of interspecies hydrogen transfer in which the mutually beneficial unidirectional transfer of hydrogen from a hydrogenproducing to a hydrogenutilising bacteria in a coupled reaction that maintains low partial pressures, which makes the transfer process hermodynamically feasible is important...... in ruminal methanogenesis. Currently, modern ‘Omics’ technologies are being applied to the study of rumen microbial ecology, genomics, metagenomics and metatranscriptomics....

Magnesium requirement of some of the principal rumen cellulolytic bacteria.

Science.gov (United States)

Morales, M S; Dehority, B A

2014-09-01

Information available on the role of Mg for growth and cellulose degradation by rumen bacteria is both limited and inconsistent. In this study, the Mg requirements for two strains each of the cellulolytic rumen species Fibrobacter succinogenes (A3c and S85), Ruminococcus albus (7 and 8) and Ruminococcus flavefaciens (B34b and C94) were investigated. Maximum growth, rate of growth and lag time were all measured using a complete factorial design, 2(3)×6; factors were: strains (2), within species (3) and Mg concentrations (6). R. flavefaciens was the only species that did not grow when Mg was singly deleted from the media, and both strains exhibited a linear growth response to increasing Mg concentrations (PR. flavefaciens B34b was estimated as 0.54 mM; whereas the requirement for R. flavefaciens C94 was >0.82 as there was no plateau in growth. Although not an absolute requirement for growth, strains of the two other species of cellulolytic bacteria all responded to increasing Mg concentrations. For F. succinogenes S85, R. albus 7 and R. albus 8, their requirement estimated from maximum growth was 0.56, 0.52 and 0.51, respectively. A requirement for F. succinogenes A3c could not be calculated because there was no solution for contrasts. Whether R. flavefaciens had a Mg requirement for cellulose degradation was determined in NH3-free cellulose media, using a 2×4 factorial design, 2 strains and 4 treatments. Both strains of R. flavefaciens were found to have an absolute Mg requirement for cellulose degradation. Based on reported concentrations of Mg in the rumen, 1.0 to 10.1 mM, it seems unlikely that an in vivo deficiency of this element would occur.

Rumen bacteria at work: bioaugmentation strategies to enhance biogas production from cow manure.

Science.gov (United States)

Ozbayram, E G; Akyol, Ç; Ince, B; Karakoç, C; Ince, O

2018-02-01

To investigate the effects of different bioaugmentation strategies for enhancing the biogas production from cow manure and evaluate microbial community patterns. Co-inoculation with cow rumen fluid and cow rumen-derived enriched microbial consortia was evaluated in anaerobic batch tests at 36°C and 41°C. Singular addition of both rumen fluid and enriched bioaugmentation culture had a promising enhancement on methane yields; however, the highest methane yield (311 ml CH 4 per gram VS at 41°C) was achieved when the anaerobic seed sludge was co-inoculated together with rumen fluid and enriched bioaugmentation culture. Bacterial community profiles were investigated by Ion PGM Platform, and specific lignocellulolytic bacteria dynamics in batch tests were assessed by qPCR. The temperature had minor effects on the abundance of bacterial community; in which Bacteroidetes and Firmicutes were the most abundant phyla in all digesters. Furthermore, Rikenellaceae, Clostridiaceae, Porphyromonadaceae, Bacteroidaceae and Ruminococcaceae played a crucial role during the anaerobic degradation of cow manure. There was an important impact of Firmicutes flavefaciens and Ruminococcus albus at 41°C, which in turn positively affected the methane production. The degree of enhancement in biogas production can be upgraded by the co-inoculation of rumen-derived bioaugmentation culture with anaerobic seed sludge with high methanogenic activity. A close look at the biotic interactions and their associations with abiotic factors might be valuable for evaluating rumen-related bioaugmentation applications. © 2017 The Society for Applied Microbiology.

A comparative study on the production rates of VFA and bacteria in the rumen of buffalo and goat estimated by isotope dilution technique

International Nuclear Information System (INIS)

Verma, D.N.; Mehra, U.R.; Singh, U.B.; Ranjhan, S.K.

1977-01-01

Digestibility trials were conducted on Murrah buffaloes and Barbari goats with rumen cannulae in the rumen to determine the digestibility of the feed constituents and the production rates of bacteria and total VFA were estimated in the rumen by isotope dilution technique. The bacterial cells growth in the rumen was more in goats than buffaloes when fed ad libitum and calculated on equal feed intake, where as, in buffaloes fed on restricted diet equal to the goats the production of bacteria and VFA were higher. Goats converted 54.04 percent of their dietary nitrogen into microbial nitrogen which was more than twice of buffaloes. (author)

Proteolytic and antimicrobial activity of lactic acid bacteria grown in goat milk.

Science.gov (United States)

Atanasova, Jivka; Moncheva, Penka; Ivanova, Iskra

2014-11-02

We examined 62 strains and 21 trade starter cultures from the collection of LB Bulgaricum PLC for proteolytic and antimicrobial activity of lactic acid bacteria (LAB) grown in goat milk. The aim of this study was to investigate the fermentation of caseins, α-lactalbumin and β-lactoglobulin by LAB, using the o -phthaldialdehyde (OPA) spectrophotometric assay and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteolysis targeted mainly caseins, especially β-casein. Whey proteins were proteolyzed, essentially β-lactoglobulin. The proteolytic activity of Lactococcus lactis l598, Streptococcus thermophilus t3D1, Dt1, Lactobacillus lactis 1043 and L. delbrueckii subsp. bulgaricus b38, b122 and b24 was notably high. The proteolysis process gave rise to medium-sized peptide populations. Most of the examined strains showed antimicrobial activity against some food pathogens, such as Escherichia coli , Staphylococcus aureus , Salmonella cholere enteridis , Listeria monocytogenes , Listeria innocua and Enterobacter aerogenes . The most active producers of antimicrobial-active peptides were strains of L. delbrueckii subsp. bulgaricus and S. thermophilus , which are of practical importance. The starter cultures containing the examined species showed high proteolytic and antimicrobial activity in skimmed goat milk. The greatest antimicrobial activity of the cultures was detected against E. aerogenes . The obtained results demonstrated the significant proteolytic potential of the examined strains in goat milk and their potential for application in the production of dairy products from goat's milk. The present results could be considered as the first data on the proteolytic capacity of strains and starter cultures in goat milk for the purposes of trade interest of LB Bulgaricum PLC.

The Cellulolytic Activity And Volatile Fatty Acid Product Of Rumen Bacteria Of Buffalo And Cattle On Rice Straw, Elephant Grass, and Sesbania Leaves Substrates

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Caribu Hadi Prayitno

1999-01-01

Full Text Available Experiment on The Cellulolytic Activity and Volatile Fatty Acid Product of Rumen Bacteria of Buffalo and Cattle on Rice Straw, Elephant Grass, and Sesbania Leaves Substrates had been conducted at Feedstuff Laboratory of Animal Science Soedirman University. The basic design  that was used in this experiment was Completely Randomized Design (CRD with factorial pattern of 6 x 3, three replications. The bacteria isolate as the factors were cellulolytic rumen bacteria isolate of buffalo (A1, A2, and A3 and cattle (A4, A5 and A6 while the substrates (second factor  were NDF rice straw (S1, elephant grass (S2, and sesbania leaves (S3 Cell walls. The result of this experiment showed that the interaction between bacteria isolate and substrate  type were significant on pH, NDF digestibility, cellulase activity, pH was  6.28 until 6.43.  The NDF digestibility range was 12.27 until 55.61 percent. The lowers of cellulase activity was 5.11 IU/ml and the higher was 24.47 IU/ml. The range of acetic acid yield was 63.37 to 307.467 mg/100 ml. Range of  propionic production was 15.17 to 352.20 mg/ 100 ml. The production of butiric acid was 8.77 to 40.87 mg/ 100 ml. The cellulase activity  of cellulolytic rumen bacteria of buffalo was higher than cattle, and also their effect on NDF digestibility of rice straw, elephant grass, and sesbania leaves cell walls. The A3 of cellulolytic rumen bacteria isolate of  buffalo changed cell walls substrat to volatile fatty  acid was more effective than cattle, especially on cell elephant grass. Propionic and butiric  acid that was produced by cellulolytic rumen bacteria isolate of buffalo more higher than cattle (Animal Production 1 (1 : 1-9 (1999 Key Words: Cellulolytic, VFA, Rumen Bacteria, Buffalo, Cattle.

Proteolytic Activity in Reduced-Fat Cheddar Cheese Made with Lactic Acid Bacteria and Camel Chymosin

DEFF Research Database (Denmark)

Børsting, Mette Winther

be the need of an extended ripening period to reach a similar cheese structure as in cheeses produced with BC. The aim of this project was to compensate for the lower proteolytic activity in cheese produced with CC compared to BC. Selection of dairy lactic acid bacteria (LAB) for cheese production with high....... lactis subsp lactis, 10 thermophilic Lactobacillus strains and 15 frozen direct vat set strains of thermopholic Lactobacillus) to hydrolyse αS1-CN, candidates were selected for cheese-making experiments. None of the selected proteolytic strains contributed significantly to softening the cheese structure...

Pathogenic Assay of Probiotic Bacteria Producing Proteolytic Enzymes as Bioremediation Bacteria Against Vannamei Shrimp Larvae (Litopenaeus vannamei

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Wilis Ari Setyati

2017-06-01

Full Text Available Application of bacteria in bioremediation of shrimp culture ponds is one of the methods used to clean internal pollutants. This study aimed to evaluate the pathogenicity of extracellular proteolytic enzyme produced by the probiotic bacteria as bioremediation bacteria on vannamei shrimp larvae culture. There were five probiotic bacteria, which were successfully isolated from the sediments served as substrate in mangrove area. The isolated bacteria were coded in number as 13, 19, 30, 33, and 36. Pathogenic bacteria Vibrio harveyi was used as positive control. Pathogenic assay was carried out in two different bacterial concentrations, i.e. 10⁸ and 10⁶ cells.mL-1. The results showed that the lowest survival rate (SR of shrimp larvae in positive control V. harveyi was 53 and 65%. Whereas isolates with the highest SR value (100% were obtained from bacteria coded as 13 and 30. Isolates no. 19, 33 and 36 had SR of more than 90%. Total plate count (TPC data showed that the bacteria increased significantly at the end of the study with an average increase value of 24%. The smallest TPC value was shown by bacterial isolate no. 19, while the largest was obtained from the isolate no. 13. These results suggest that all probiotic bacteria were not pathogenic to the vannamei shrimp larvae.   Keywords: aquaculture, shrimp, bioremediation, pathogenesis, vibrio.

PROTEOLYTIC AND FIBRINOLYTIC ACTIVITIES OF HALOPHILIC LACTIC ACID BACTERIA FROM TWO INDONESIAN FERMENTED FOODS

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Asep A. Prihanto

2013-04-01

Full Text Available Exploration of fermented foods as sources of fibrinolytic enzymes is increased in the last decades. Terasi and Jambal roti is Indonesian traditional fermented fish products, which were famous in Java Island. Both are important products in Indonesian dishes, especially in Java. Investigation on halophilic lactic acid bacteria using MRS and M-17 agar obtained seventy four isolated strains. Their proteolytic and fibrinolytic activities were determined using skim milk agar and plasminogen-free fibrin plate. Twenty five isolates showed protease activities, while only four of them secreted fibrinolitic enzyme. The highest proteolytic and fibrinolytic activity was shown by TB1 strain, which is identified as Bacillus coagulans. The 16s rDNA is still in investigating to confirm the TB1 strain identity.

Short communication: Intestinal digestibility of amino acids in fluid- and particle-associated rumen bacteria determined using a precision-fed cecectomized rooster bioassay.

Science.gov (United States)

Fonseca, A C; Fredin, S M; Ferraretto, L F; Parsons, C M; Utterback, P L; Shaver, R D

2014-01-01

Microbial protein represents the majority of metabolizable protein absorbed by ruminant animals. Enhanced understanding of the AA digestibility of rumen microbes will improve estimates of metabolizable protein. The objective of this experiment was to determine the digestibility of AA in fluid- (FAB) and particle-associated bacteria (PAB) using the precision-fed cecectomized rooster bioassay. Bacteria were isolated from 4 ruminally cannulated lactating Holstein cows by differential centrifugation, including particle suspension in 0.1% Tween-80 for increased removal of PAB from ruminal digesta. Samples of FAB and PAB were fed to 9 cecectomized roosters to determine standardized digestibility of AA. Total AA digestibility was 76.8 and 75.5% for FAB and PAB, respectively, but did not differ. Differences existed in AA digestibilities within bacterial type when compared with the mean essential AA digestibility value. Compared with previous literature estimates of AA digestibility in microbes (mean = 76%; range = 57-87%) and relative to National Research Council estimates of total AA from rumen bacteria (80%), the precision-fed cecectomized rooster assay is an acceptable in vivo model to determine AA digestibility of rumen bacteria. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Horizontal gene transfer from Bacteria to rumen Ciliates indicates adaptation to their anaerobic, carbohydrates-rich environment.

NARCIS (Netherlands)

Ricard, G.N.S.; McEwan, N.R.; Dutilh, B.E.; Jouany, J.P.; Macheboeuf, D.; Mitsumori, M.; McIntosh, F.M.; Michalowski, T.; Nagamine, T.; Nelson, N.; Newbold, C.J.; Nsabimana, E.; Takenaka, A; Thomas, N.A.; Ushida, K.; Hackstein, J.H.P.; Huynen, M.A.

2006-01-01

BACKGROUND: The horizontal transfer of expressed genes from Bacteria into Ciliates which live in close contact with each other in the rumen (the foregut of ruminants) was studied using ciliate Expressed Sequence Tags (ESTs). More than 4000 ESTs were sequenced from representatives of the two major

Measurement of the rate of production of bacteria in the rumen of buffalo calves using 14C Str. Bovis

International Nuclear Information System (INIS)

Verma, D.N.; Singh, U.B.; Srivastava, S.K.; Srivastava, R.V.N.

1976-01-01

A technique has been developed for the in vivo estimation of the rate of production of bacteria in the rumen of buffalo calves. Three male buffalo calves (Des bubalis) of about 2 years of age were taken for these experiments. The animals were given 20 kg. of green maize in 12 equal amounts at 2 hourly intervals. The streptococcus bovis isolated from the rumen were labelled with 14 C by in vitro incubation in the presence of (U- 14 C) DL-leucine. Labelled streptococcus bovis were injected in a single dose in the rumen. Tracer dilution kinetics for open system were applied to the data obtained as a result of dilution of Str. bovis in the rumen to obtained production rates. The average turnover time and production rates were 424.54 minute and 87.43mg/minute. (author)

Metagenomic analysis of bacterial community structure and diversity of lignocellulolytic bacteria in Vietnamese native goat rumen.

Science.gov (United States)

Do, Thi Huyen; Dao, Trong Khoa; Nguyen, Khanh Hoang Viet; Le, Ngoc Giang; Nguyen, Thi Mai Phuong; Le, Tung Lam; Phung, Thu Nguyet; van Straalen, Nico M; Roelofs, Dick; Truong, Nam Hai

2018-05-01

In a previous study, analysis of Illumina sequenced metagenomic DNA data of bacteria in Vietnamese goats' rumen showed a high diversity of putative lignocellulolytic genes. In this study, taxonomy speculation of microbial community and lignocellulolytic bacteria population in the rumen was conducted to elucidate a role of bacterial structure for effective degradation of plant materials. The metagenomic data had been subjected into Basic Local Alignment Search Tool (BLASTX) algorithm and the National Center for Biotechnology Information non-redundant sequence database. Here the BLASTX hits were further processed by the Metagenome Analyzer program to statistically analyze the abundance of taxa. Microbial community in the rumen is defined by dominance of Bacteroidetes compared to Firmicutes. The ratio of Firmicutes versus Bacteroidetes was 0.36:1. An abundance of Synergistetes was uniquely identified in the goat microbiome may be formed by host genotype. With regard to bacterial lignocellulose degraders, the ratio of lignocellulolytic genes affiliated with Firmicutes compared to the genes linked to Bacteroidetes was 0.11:1, in which the genes encoding putative hemicellulases, carbohydrate esterases, polysaccharide lyases originated from Bacteroidetes were 14 to 20 times higher than from Firmicutes. Firmicutes seem to possess more cellulose hydrolysis capacity showing a Firmicutes/Bacteroidetes ratio of 0.35:1. Analysis of lignocellulolytic potential degraders shows that four species belonged to Bacteroidetes phylum, while two species belonged to Firmicutes phylum harbouring at least 12 different catalytic domains for all lignocellulose pretreatment, cellulose, as well as hemicellulose saccharification. Based on these findings, we speculate that increasing the members of Bacteroidetes to keep a low ratio of Firmicutes versus Bacteroidetes in goat rumen has resulted most likely in an increased lignocellulose digestion.

AKTIVITAS PROTEOLITIK BAKTERI ASAM LAKTAT DALAM FERMENTASI SUSU KEDELAI [Proteolytic Activities of Lactic Acid Bacteria in Fermentation of Soymilk

OpenAIRE

Yusmarini1,2)*; R. Indrati1); T. Utami1); Y. Marsono1)

2010-01-01

Some lactic acid bacteria (LAB) strains had been isolated from spontaneously fermented soymilk which have proteolytic system. The purpose of this research was to study ability of isolates in fermentation of soymilk. The changes in bacterial growth, pH, titrable acidity, and proteolytic activities during fermentation were examined. Isolates of Lactobacillus plantarum 1 R.1.3.2; L. plantarum 1 R.11.1.2 and L. acidophilus FNCC 0051 (as a control) were capable growing in soymilk. The results indi...

Isolation and identification of acetogenic bacteria obtained from deer rumen and their potential for methanogenesis inhibitor

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Amlius Thalib

2008-10-01

Full Text Available Methanogenesis can be inhibited by various chemicals through different mechanism reaktion. The use of acetogenic bacteria as H2 sink is assumed to be a promising approach. Isolation and identification of acetogenic bacteria obtained from deer rumen had been conducted. Two types of media used for isolation were hydrogen-carbondioxide utilizing acetogens and carbonmonoxide utilizing acetogens. Identification of species of acetogens isolates was based on descriptions of morphology, Gram type, motility, bioreaction results, and oksygen requirement. The compositions of methane and volatile fatty acids (VFA were determined on minimal media or added with sheep rumen liquid innoculated with pure isolates. The identification results showed that the isolate cultured on media of hydrogen-carbondioxide utilizing acetogens was Acetoanaerobium noterae and the ones cultured on media of carbonmonoxide utilizing acetogens was Acetobacterium woodii. Inoculumn of A. noterae and A. woodii could decreased the composition of methane resulted from substrate fermented by fresh rumen liquid of sheep (CRDF, that is culture of A. noterae added FPM and defaunator decreased methane production by 28.8% (P CH3COOH + 2H2O by which reduction of CO2 with H2 producing CH4 can be inhibited or decreased. Their function as methanogenesis inhibitor would be more significant when they are combined with microbial growth factors and defaunator.

Variability of Actinobacteria, a minor component of rumen microflora.

Science.gov (United States)

Suľák, M; Sikorová, L; Jankuvová, J; Javorský, P; Pristaš, P

2012-07-01

Actinobacteria (Actinomycetes) are a significant and interesting group of gram-positive bacteria. They are regular, though infrequent, members of the microbial life in the rumen and represent up to 3 % of total rumen bacteria; there is considerable lack of information about ecology and biology of rumen actinobacteria. During the characterization of variability of rumen treponemas using non-cultivation approach, we also noted the variability of rumen actinobacteria. By using Treponema-specific primers a specific 16S rRNA gene library was prepared from cow and sheep rumen total DNA. About 10 % of recombinant clones contained actinobacteria-like sequences. Phylogenetic analyses of 11 clones obtained showed the high variability of actinobacteria in the ruminant digestive system. While some sequences are nearly identical to known sequences of actinobacteria, we detected completely new clusters of actinobacteria-like sequences, representing probably new, as yet undiscovered, group of rumen Actinobacteria. Further research will be necessary for understanding their nature and functions in the rumen.

Defined bacterial populations in the rumens of gnotobiotic lambs.

Science.gov (United States)

Lysons, R J; Alexander, T J; Wellstead, P D; Hobson, P N; Mann, S O; Stewart, C S

1976-06-01

Five gnotobiotic lambs were fed on sterile diets until they were killed at 13 to 21 weeks of age. They were dosed orally with different combinations of 11 species of rumen bacteria. The biochemical reactions of each of the bacteria inoculated had been determined in pure culture in vitro, and they were chosen to perform the main reactions known to be associated with digestion in the normal mature rumen. Two of the bacteria could not be reisolated, but the remainder had established readily in the rumen, forming stable, mixed, defined populations. The total numbers of bacteria in the rumen, and the viable counts of most of the individual species were comparable to those of normal sheep. The concentration of volatile fatty acids was lower, however, and in four of the lambs there was a higher proportion of butyric acid and a lower proportion of propionic acid than in normal sheep. Cellulolytic, ureolytic, and methanogenic activities appeared to be taking place and lactate-utilizing bacteria appeared to reverse the accumulation of lactate which resulted from the activity of lactate-producing bacteria. Some of the bacteria also established at high levels in the caecum.

Effect of the yeast and bacteria biomass on the microbiota in the rumen.

Science.gov (United States)

Vamanu, E; Vamanu, A; Popa, O; Vassu, Tatiana; Ghindea, Raluca; Pelinescu, Diana; Nita, Sultana; Babeanu, Narcisa

2008-09-15

This study aims at obtaining a probiotic product based on viable biomass from 6 yeast strains and 2 strains of lactic bacteria used for nutrition of animals. The strains are subjected to some resistance tests, at temperature, pH, pepsin, pancreatin and biliary salts so as to make obvious their viability. Tests were done by comparison to the witness strain and respectively a protective solution based on mucin and casein. Based on the resulted viabilities 2 products are formulated. Their effect is tested by inoculating fresh rumen content and supervising the microbic balance for a period of 12 days. After the final tests, it resulted that the product Fpl (20% Saccharomyces cerevisiae 1-29, 10% Kluyveromyces marxianus R-CS, 20% Issatchenkia orientalis R-BC, 30% Lactobacillus paracasei CMGB16, 20% Enterococcus faecium GM8) was chosen because anaerobic strains were preponderant as a consequence of the tests performed with rumen.

EFECTO DE LA PROPORCIÓN CONCENTRADO / FORRAJE DE LA DIETA, SOBRE LAS POBLACIONES DE BACTERIAS UTILIZADORAS DE HIDRÓGENO DEL RUMEN

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Alcantar Díaz BE

2010-07-01

Full Text Available Las Archae metanogénicas, las bacterias Acetogénicas y Sulfato reductoras son los tres grupos últimos de microorganismos dentro de la cadena de degradación de los sustratos en el ecosistema del rumen y la manipulación del tamaño de estas poblaciones es benéfica para la maximización de la energía en el rumiante. Para evaluar el efecto de dietas con diferente proporción de forraje/concentrado, sobre el número de bacterias utilizadoras de hidrógeno en el rumen in vivo, 16 ovino machos fueron colocados en cajas metabólicas y se alimentaron con cuatro dietas a base forraje de maíz más con- centrado en las proporciones: C20, C40, C60 y C80. El concentrado fue a base de maíz molido, harina de soya y trigo. Se tomaron muestras de contenido del rumen cada dos horas durante 24 horas, para determinar número de bacterias, concentraciones de acetato, propionato, amonio y pH. El número de bacterias se determinó por el método del número más probable. El diseño en cuadro latino fue para evaluar el efecto del factor A=4 dietas, el factor B los tiempos de incubación, el factor C cuatro repeticiones y sus interacciones. La proporción forraje: concentrado de la dieta, muestra diferencia significativas (p<0.001 entre el número de bacterias metanogénicas y acetogénicas y sulfato reductoras. Las dietas C60 y C80, tienen el mayor número de bacterias acetogénicas. El efecto de la dieta y tiempo de fermentación, también presentó diferencias en los valores de acetato (p<0.0006, propionato (p< 0.0001, amonio (p<0.0001 y pH (p<0.0001. La manipulación de la proporción forraje: concentrado de la dieta usada en la alimentación de rumiantes es una estrategia para establecer determinado grupo de bacterias utilizadoras de hidrogeno y las propiedades del ambiente en el rumen.

Formation of methylamine by rumen microorganisms

International Nuclear Information System (INIS)

Itabashi, Hisao; Kandatsu, Makoto.

1978-01-01

An unknown ninhydrin positive peak on the chromatograms of amino acid analyzer of alkalified rumen fluid distillate of goats was isolated as DNP-derivative and identified as methylamine. Under normal feeding condition, its concentration in the rumen ranged 0.1-3.9 mgN/100 ml of rumen fluid and the proportion of methylamine in total volatile base, or apparent ammonia, ranged 0.5-13% during post-feeding. When ammonium salt was administered into the rumen with hay-concentrate ration, these values were increased up to 8.1 mgN/100 ml and 25.8% respectively. Concentrations of ammonia and methylamine when aspartic acid or alanine was administered into the rumen in place of concentrate mixture (control) were not markedly different from the control. In the case of arginine, glutamic acid or glycine administration, these concentrations were depressed as compared to the control. There were no distinct differences in the concentration of methylamine between the faunated and unfaunated goats. 14 C from 14 C-chlorella protein hydrolyzates, U- 14 C-alanine, 2- 14 C-glycine or 14 C-sodium bicarbonate was incorporated into methylamine in invitro incubation with rumen micro-organisms. When the washed suspensions of rumen bacteria or protozoa were incubated with 14 C-chlorella protein hydrolyzates, the radioactivity in methylamine appeared only in the case of bacteria suspensions. After the addition of 15 N-ammonium citrate into the rumen, the incorporation of 15 N into methylamine was observed during 1-9 hr. (auth.)

Anaerobic degradation of veratrylglycerol-beta-guaiacyl ether and guaiacoxyacetic acid by mixed rumen bacteria.

OpenAIRE

Chen, W; Supanwong, K; Ohmiya, K; Shimizu, S; Kawakami, H

1985-01-01

Veratrylglycerol-beta-guaiacyl ether (0.2 g/liter), a lignin model compound, was found to be degraded by mixed rumen bacteria in a yeast extract medium under strictly anaerobic conditions to the extent of 19% within 24 h. Guaiacoxyacetic acid, 2-(o-methoxyphenoxy)ethanol, vanillic acid, and vanillin were detected as degradation products of veratrylglycerol-beta-guaiacyl ether by thin-layer chromatography, gas chromatography, and gas chromatography-mass spectrometry. Guaiacoxyacetic acid (0.25...

High-grain diets altered rumen fermentation and epithelial bacterial community and resulted in rumen epithelial injuries of goats.

Science.gov (United States)

Zhang, Ruiyang; Ye, Huimin; Liu, Junhua; Mao, Shengyong

2017-09-01

This study evaluated the effects of high-grain diets on the rumen fermentation, epithelial bacterial community, morphology of rumen epithelium, and local inflammation of goats during high-grain feeding. Twelve 8-month-old goats were randomly assigned to two different diets, a hay diet or a high-grain diet (65% grain, HG). At the end of 7 weeks of treatment, samples of rumen content and rumen epithelium were collected. Rumen pH was lower (P rumen epithelial bacterial community, with an increase in the proportion of genus Prevotella and a decrease in the relative abundance of the genera Shuttleworthia and Fibrobacteres. PICRUSt analysis suggested that the HG-fed group had a higher (P rumen epithelial injury and upregulated (P rumen pH, LPS level, and rumen epithelial bacteria abundance. In conclusion, our results indicated that the alterations in the rumen environment and epithelial bacterial community which were induced by HG feeding may result in the damage and local inflammation in the rumen epithelium, warranting further study of rumen microbial-host interactions in the HG feeding model.

Effect of Carbohydrate Sources and Levels of Cotton Seed Meal in Concentrate on Feed Intake, Nutrient Digestibility, Rumen Fermentation and Microbial Protein Synthesis in Young Dairy Bulls

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M. Wanapat

2013-04-01

Full Text Available The objective of this study was to investigate the effect of levels of cottonseed meal with various carbohydrate sources in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in dairy bulls. Four, 6 months old dairy bulls were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC and cassava chip+rice bran in the ratio of 3:1 (CR3:1, and factor B was cotton seed meal levels in the concentrate; 109 g CP/kg (LCM and 328 g CP/kg (HCM at similar overall CP levels (490 g CP/kg. Bulls received urea-lime treated rice straw ad libitum and were supplemented with 10 g of concentrate/kg BW. It was found that carbohydrate source and level of cotton seed meal did not have significant effects on ruminal pH, ammonia nitrogen concentration, microbial protein synthesis or feed intake. Animals which received CC showed significantly higher BUN concentration, ruminal propionic acid and butyric acid proportions, while dry matter, organic matter digestibility, populations of total viable bacteria and proteolytic bacteria were lower than those in the CR3:1 treatment. The concentration of total volatile fatty acids was higher in HCM than LCM treatments, while the concentration of butyric acid was higher in LCM than HCM treatments. The population of proteolytic bacteria with the LCM treatments was higher than the HCM treatments; however other bacteria groups were similar among the different levels of cotton seed meal. Bulls which received LCM had higher protein digestibility than those receiving HCM. Therefore, using high levels of cassava chip and cotton seed meal might positively impact on energy and nitrogen balance for the microbial population in the rumen of the young dairy bull.

AKTIVITAS PROTEOLITIK BAKTERI ASAM LAKTAT DALAM FERMENTASI SUSU KEDELAI [Proteolytic Activities of Lactic Acid Bacteria in Fermentation of Soymilk

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Yusmarini1,2*

2010-12-01

Full Text Available Some lactic acid bacteria (LAB strains had been isolated from spontaneously fermented soymilk which have proteolytic system. The purpose of this research was to study ability of isolates in fermentation of soymilk. The changes in bacterial growth, pH, titrable acidity, and proteolytic activities during fermentation were examined. Isolates of Lactobacillus plantarum 1 R.1.3.2; L. plantarum 1 R.11.1.2 and L. acidophilus FNCC 0051 (as a control were capable growing in soymilk. The results indicated that initial pH of soymilk was 6,6 and decreased to 4,6 after fermentation and titrable acidity of 0.11 increased to 0.34 after fermentation. The proteolytic activities were 0.352 U/ml – 0.468 U/ml. The electrophoretic pattern of the proteins showed changes during fermentation of soymilk.

Amino acid composition of rumen bacteria and protozoa in cattle.

Science.gov (United States)

Sok, M; Ouellet, D R; Firkins, J L; Pellerin, D; Lapierre, H

2017-07-01

Because microbial crude protein (MCP) constitutes more than 50% of the protein digested in cattle, its AA composition is needed to adequately estimate AA supply. Our objective was to update the AA contributions of the rumen microbial AA flowing to the duodenum using only studies from cattle, differentiating between fluid-associated bacteria (FAB), particle-associated bacteria (PAB), and protozoa, based on published literature (53, 16, and 18 treatment means were used for each type of microorganism, respectively). In addition, Cys and Met reported concentrations were retained only when an adequate protection of the sulfur groups was performed before the acid hydrolysis. The total AA (or true protein) fraction represented 82.4% of CP in bacteria. For 10 AA, including 4 essential AA, the AA composition differed between protozoa and bacteria. The most noticeable differences were a 45% lower Lys concentration and 40% higher Ala concentration in bacteria than in protozoa. Differences between FAB and PAB were less pronounced than differences between bacteria and protozoa. Assuming 33% FAB, 50% PAB, and 17% of protozoa in MCP duodenal flow, the updated concentrations of AA would decrease supply estimates of Met, Thr, and Val originating from MCP and increase those of Lys and Phe by 5 to 10% compared with those calculated using the FAB composition reported previously. Therefore, inclusion of the contribution of PAB and protozoa to the duodenal MCP flow is needed to adequately estimate AA supply from microbial origin when a factorial method is used to estimate duodenal AA flow. Furthermore, acknowledging the fact that hydrolysis of 1 kg of true microbial protein yields 1.16 kg of free AA substantially increases the estimates of AA supply from MCP. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

PCR-DGGE Analysis of Bacterial Population Attached to the Bovine Rumen Wall

OpenAIRE

Lukáš, F. (Filip); Šimůnek, J. (Jiří); Mrázek, J. (Jakub); Kopečný, J. (Jan)

2010-01-01

We isolated and amplified by PCR 16S rDNA from bacteria attached to the bovine rumen wall and analyzed it by denaturing gradient gel electrophoresis (DGGE) with subsequent sequence analysis. The attached bacterial community differed from the bacteria of rumen content; however, no differences were observed among the five epithelial sampling sites taken from each animal. The DGGE profile of the bacterial population attached to the rumen wall represented a high inter-animal variation.

The influence of radiation on bacterial cells and their proteolytic properties

International Nuclear Information System (INIS)

Szulc, M.; Stefaniakowa, A.; Stanczak, B.; Peconek, J.

1980-01-01

The suspension of bacterial cells and their spores were exposed to X rays in the environment with and without protein. The doses of radiation ranged from 1 to 100 Gy and in case of spores of B. subtilis from 50 to 1000 Gy. It was found that irradiation to Proteus vulgaris, Pseudomonas fluorescens and Ps. aeruginosa caused an inconsiderable decrease of proteolytic properties of the generation originated from irradiated bacteria. Irradiation of B. subtilis spores did not influence the proteolytic activity of bacterial cells derived from the exposed spores. The degree of wasting away of bacteria exposed to the same radiation was higher than the rate of proteolytic properties decrease. The presence of protein in the surroundings had no influence on proteolytic characteristics of new generations. (author)

Evaluation of sonication treatment and buffer composition on rumen bacteria protein extraction and carboxymethylcellulase activity.

Science.gov (United States)

Prauchner, Carlos A; Kozloski, Gilberto V; Farenzena, Roberta

2013-05-01

The methodological procedures for studying the fibrolytic activity of rumen bacteria are not clearly established. In this study the efficiency of sonication treatment and buffer composition (i.e. buffer varying in tonicity or pH) on the level of protein extraction from the residue of forage samples incubated in the rumen of a grazing steer and the effect of buffer composition or CaCl₂ concentration on the carboxymethylcellulase (CMCase) activity of the released protein were evaluated. The amount of protein released from the residue of incubation was higher (P buffer pH (P buffer. Both linear and quadratic effects (P buffer pH on CMCase activity were significant, with CMCase activity being maximal at pH 5.4-6.1. CMCase activity was higher (P buffer solution that includes a calcium source. When pH is not a treatment factor, the buffer pH should be between 5.5 and 6. © 2012 Society of Chemical Industry.

Microbiological Monitoring and Proteolytic Study of Clinical Samples From Burned and Burned Wounded Patients

International Nuclear Information System (INIS)

Toema, M.A.; El-Bazza, Z.E.; El-Hifnawi, H.N.; Abd-El-Hakim, E.E.

2013-01-01

In this study, clinical samples were collected from 100 patients admitted to Burn and Plastic Surgery Department, Faculty of Medicine, Ain Shams University, Egypt, over a period of 12 months. The proteolytic activity of 110 clinical samples taken from surfaces swabs which taken from burned and burned wounded patients with different ages and gender was examined. Screening for the proteolytic activity produced by pathogenic bacteria isolated from burned and burned wounded patients was evaluated as gram positive Bacilli and gram negative bacilli showed high proteolytic activity (46.4%) while 17.9% showed no activity. The isolated bacteria proved to have proteolytic activity were classified into high, moderate and weak. The pathogenic bacteria isolated from burned and burned wounded patients and showing proteolytic activity were identified as Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris, Bacillus megaterium, Bacillus cereus, Staphylococcus aureus, Escherichia coli, Klebsiella ozaeanae, Klebsiella oxytoca, Klebsiella pneumoniae and Pseudomonas fluoresces.

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

Science.gov (United States)

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 ...

Genomes of rumen bacteria encode atypical pathways for fermenting hexoses to short-chain fatty acids.

Science.gov (United States)

Hackmann, Timothy J; Ngugi, David Kamanda; Firkins, Jeffrey L; Tao, Junyi

2017-11-01

Bacteria have been thought to follow only a few well-recognized biochemical pathways when fermenting glucose or other hexoses. These pathways have been chiseled in the stone of textbooks for decades, with most sources rendering them as they appear in the classic 1986 text by Gottschalk. Still, it is unclear how broadly these pathways apply, given that they were established and delineated biochemically with only a few model organisms. Here, we show that well-recognized pathways often cannot explain fermentation products formed by bacteria. In the most extensive analysis of its kind, we reconstructed pathways for glucose fermentation from genomes of 48 species and subspecies of bacteria from one environment (the rumen). In total, 44% of these bacteria had atypical pathways, including several that are completely unprecedented for bacteria or any organism. In detail, 8% of bacteria had an atypical pathway for acetate formation; 21% of bacteria had an atypical pathway for propionate or succinate formation; 6% of bacteria had an atypical pathway for butyrate formation and 33% of bacteria had an atypical or incomplete Embden-Meyerhof-Parnas pathway. This study shows that reconstruction of metabolic pathways - a common goal of omics studies - could be incorrect if well-recognized pathways are used for reference. Furthermore, it calls for renewed efforts to delineate fermentation pathways biochemically. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Potency of fiber rumen bacterial isolates from local buffalo inoculated into Frisian Holstein calves during preweaning period

Directory of Open Access Journals (Sweden)

Iwan Prihantoro

2012-12-01

Full Text Available Fiber-digesting bacteria are the main rumen bacteria that play an important role in digesting feed. These bacteria are adapted to low quality forage from agricultural byproduct. The aim of these study was to determine the potency of fiber-digesting bacteria consortium obtained from buffalo rumen inoculated to Frisian Holstein calves during preweaning on feed consumption, utilization, mineral uptake and physiological status. This study used 14 isolates of bacteria obtained from collection of Faculty of Animal Science, Bogor Agricultural University. The experimental unit consisted of six Frisian Holstein calves at two week old with the average body weight of 38.00 ± 6.23 kg. Calves were inoculated by 20 ml of fiber-digesting rumen bacterial isolates [4.56 x 109 cfu/ml] every morning for four weeks. Experimental design used was based on a completly randomized design with three calves received the respective inoculation (treatment group and three calves without any inoculation (control group. Data were analyzed statistically using t-test method with α = 0.05 and 0.01. The results showed that fiber-digesting bacteria (FDB from rumen buffalo have adapted in the calves rumen since preweaning periode. Inoculation FDB increased the number of rumen bacteria, digestibility of protein and P uptake calves at eight weeks old. Increased feed intake, uptake of Mg and cobalt calves at 14 weeks old. Without causing any negative effects on ADG, physiological status and rumen fermentability.

Screening of Lactic Acid Bacteria from Rumen Liquor and King Grass Silage as well as Their Antibacterial Activities

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A. Sofyan

2013-12-01

Full Text Available Probiotic is a live microbial culture which has positive effect on animal by improving the natural balance of microflora in the digestive tract. This experiment aimed to screen and identify indigenous lactic acid bacteria (LAB from rumen liquor and king grass (Pennisetum hybrid silage as a probiotic candidate and to evaluate their resistance in low pH, and inhibitory activities against pathogenic bacteria. The LAB isolate was characterized by a clear zone formed on MRSA medium + CaCO3 0.2% (w/v and further identified by morphological and biochemical assays. The selected isolates were evaluated for their viability in low pH, pathogenic bacterial inhibition, and lactic acid production. The experimental arrangement was a factorial block design (4 x 2 consisted of four isolates and two levels of pH value (pH 2 and 3, each treatment in 3 equal replicates. The result showed that four isolates (two isolates from the rumen liquor of fistulated cattle and two isolates from silage were identified as lactic acid bacteria. The four isolates showed inhibition activity against Escherichia coli, Bacillus subtilis, Staphylococcus aureus and performed viability at low pH during 2 h treatment. The highest lactic acid production was obtained from isolates Sil.3 (21.42% and followed by CR2 (19.88%, CR1 (15.40% and Sil.9 (15.08%. Biochemical identification by standard of analytical profile index (API 50 CHL kit showed that the selected isolates CR1 was Lactobacillus paracasei ssp. paracasei 3 (91.5%, L. paracasei ssp. paracasei 3 (76.5%, Sil.3 was Lactobacillus brevis (95.1%, and Sil.9 was Lactobacillus collinoides (92.5%. In conclusion, probiotic candidates isolated from rumen liquor are confirmed as L. paracasei ssp. paracasei (CR1 and CR2, while two other isolates from king grass silage are identified as L. brevis (Sil.3 and L. collinoides (Sil.9. L. brevis (Sil.3 and L. paracasei ssp. paracasei (CR1 has higher inhibition against pathogenic bacteria (E. coli, S

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

Science.gov (United States)

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). 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.

Genomes of rumen bacteria encode atypical pathways for fermenting hexoses to short-chain fatty acids

KAUST Repository

Hackmann, Timothy J.

2017-09-11

Bacteria have been thought to follow only a few well-recognized biochemical pathways when fermenting glucose or other hexoses. These pathways have been chiseled in the stone of textbooks for decades, with most sources rendering them as they appear in the classic 1986 text by Gottschalk. Still, it is unclear how broadly these pathways apply, given that they were established and delineated biochemically with only a few model organisms. Here we show that well-recognized pathways often cannot explain fermentation products formed by bacteria. In the most extensive analysis of its kind, we reconstructed pathways for glucose fermentation from genomes of 48 species and subspecies of bacteria from one environment (the rumen). In total, 44% of these bacteria had atypical pathways, including several that are completely unprecedented for bacteria or any organism. In detail, 8% of bacteria had an atypical pathway for acetate formation; 21% for propionate or succinate formation; 6% for butyrate formation; and 33% had an atypical or incomplete Embden-Meyerhof-Parnas pathway. This study shows that reconstruction of metabolic pathways-a common goal of omics studies-could be incorrect if well-recognized pathways are used for reference. Further, it calls for renewed efforts to delineate fermentation pathways biochemically. This article is protected by copyright. All rights reserved.

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

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N. Anantasook

2013-11-01

Full Text Available 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.

Ammonia Production by Ruminal Microorganisms and Enumeration, Isolation, and Characterization of Bacteria Capable of Growth on Peptides and Amino Acids from the Sheep Rumen

OpenAIRE

Eschenlauer, S. C. P.; McKain, N.; Walker, N. D.; McEwan, N. R.; Newbold, C. J.; Wallace, R. J.

2002-01-01

Excessive NH3 production in the rumen is a major nutritional inefficiency in ruminant animals. Experiments were undertaken to compare the rates of NH3 production from different substrates in ruminal fluid in vitro and to assess the role of asaccharolytic bacteria in NH3 production. Ruminal fluid was taken from four rumen-fistulated sheep receiving a mixed hay-concentrate diet. The calculated rate of NH3 production from Trypticase varied from 1.8 to 19.7 nmol mg of protein−1 min−1 depending on...

Insights into the bovine rumen plasmidome

Science.gov (United States)

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

2012-01-01

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

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

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Robert W Li

Full Text Available 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.

Effects of Plant Herb Combination Supplementation on Rumen Fermentation and Nutrient Digestibility in Beef Cattle

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M. Wanapat

2013-08-01

Full Text Available Four rumen-fistulated crossbred beef cattle (Brahman native were randomly assigned according to a 4×4 Latin square design experiment to be fed plant herb supplements in their concentrate mixture. The treatments were: without herb supplementation (Control, lemongrass meal supplementation at 100 g/d (L, lemongrass meal supplementation at 100 g/d plus peppermint powder at 10 g/d (LP, and lemongrass meal supplementation at 100 g/d plus peppermint powder at 10 g/d with garlic powder 40 g/d (LPG, respectively. Based on the present study, the DMI and apparent digestibility of DM, OM, aNDF and ADF were not affected by dietary herb supplementation while CP digestibility tended to be decreased by herb supplement. Moreover, NH3-N and BUN were decreased in all herb supplemented treatments and there was a tendency to an increase in ruminal pH in all herb supplemented groups. While there was no change in TVFA and C4 among lemongrass treatments, C2 was decreased in all herb supplemented treatments while C3 was increased. Methane production by calculation was the lowest in the LP and LPG groups. Population sizes of bacteria and protozoa were decreased in all herb supplemented groups, but not fungal zoospores. In all supplemented groups, total viable and proteolytic bacteria were decreased, while amylolytic and cellulolytic bacteria were similar. More importantly, in all herb supplemented groups, there were higher N balances, while there was no difference among treatments on purine derivative (PD excretion or microbial N. Based on the results above, it could be concluded that there was no negative effect on ruminal fermentation characteristics and nutrient utilization by plant herb supplement, but protozoal population and CH4 production were reduced. Thus, lemongrass alone or in combination with peppermint and garlic powder could be used as feed additives to improve rumen fermentation efficiency.

Effects of alternative protein sources on rumen microbes and productivity of dairy cows

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Metha Wanapat

2011-01-01

Full Text Available This experiment was conducted to investigate the effect of various protein sources on digestibility, rumen fermentation, milk yield and milk composition in dairy cows. Four Holstein Friesian native crossbred cows in early lactating were randomly assigned according to a 4x4 Latin square design. The dietary treatments containing different protein sources in concentrate diets were soybean meal (SBM, cassava hay (CH, Leucaena leucocephala (LL and yeast-fermented cassava chips (YEFECAP, with ad libitum intake of urea-treated rice straw. Digestibility of DM, OM, NDF and ADF was not different among treatments (P>0.05 while CP digestibility was highest (P<0.05 in CH and YEFECAP supplemented groups. Ruminal NH3-N and BUN concentrations varied among protein sources and were highest in SBM and LL fed groups (P<0.05. Ruminal total volatile fatty acid (VFA and propionic acid were found highest in cows receiving CH and YEFECAP (P<0.05. Ruminal fungi, proteolytic and cellulolytic bacteria were highest when YEFECAP was supplemented. Milk fat and milk protein were significantly increased (P<0.05 in cows fed with CH and YEFECAP. Based on this study, it was concluded that providing CH or YEFECAP as protein source in concentrate diets could improve rumen fermentation and milk production in lactating dairy cows fed on rice straw.

Omics insights into rumen ureolytic bacterial community and urea metabolism in dairy cows

OpenAIRE

Jin, Di

2017-01-01

Urea has been used in the diets of ruminants as a non-protein nitrogen source. Ureolytic bacteria are key organisms in the rumen producing urease enzymes to catalyze the breakdown of urea to ammonia (NH3), and the NH3 is used as nitrogen for microbial protein synthesis. In the rumen, hydrolysis of urea to NH3 occurs at a greater rate than NH3 can be utilized by rumen bacteria, and excess ammonia absorbed into blood may be harmful to the animals. Nowadays, little is known about the information...

Diversity of phytases in the rumen.

Science.gov (United States)

Nakashima, Brenda A; McAllister, Tim A; Sharma, Ranjana; Selinger, L Brent

2007-01-01

Examples of a new class of phytase related to protein tyrosine phosphatases (PTP) were recently isolated from several anaerobic bacteria from the rumen of cattle. In this study, the diversity of PTP-like phytase gene sequences in the rumen was surveyed by using the polymerase chain reaction (PCR). Two sets of degenerate primers were used to amplify sequences from rumen fluid total community DNA and genomic DNA from nine bacterial isolates. Four novel PTP-like phytase sequences were retrieved from rumen fluid, whereas all nine of the anaerobic bacterial isolates investigated in this work contained PTP-like phytase sequences. One isolate, Selenomonas lacticifex, contained two distinct PTP-like phytase sequences, suggesting that multiple phytate hydrolyzing enzymes are present in this bacterium. The degenerate primer and PCR conditions described here, as well as novel sequences obtained in this study, will provide a valuable resource for future studies on this new class of phytase. The observed diversity of microbial phytases in the rumen may account for the ability of ruminants to derive a significant proportion of their phosphorus requirements from phytate.

Bacterial community in the rumen of Tibetan sheep and Gansu alpine fine-wool sheep grazing on the Qinghai-Tibetan Plateau, China.

Science.gov (United States)

Huang, Jinqiang; Li, Yongjuan; Luo, Yuzhu

2017-05-12

The rumen microbiome plays a vital role in ruminant nutrition and health, and its community is affected by environmental factors. However, little is known about the rumen bacterial community of ruminants living in the special ecological environment of the Qinghai-Tibetan Plateau (QTP) of China. The objectives of this study were to investigate the rumen bacterial community of the typical plateau sheep (Tibetan sheep, TS, and Gansu alpine fine-wool sheep, GS) grazing on the QTP, using 16S rRNA gene sequence analysis, and to evaluate the relationship between the rumen bacterial community and the QTP environment. A total of 116 sequences (201 clones) were examined and divided into 53 operational taxonomic units (OTUs) in the TS library and 46 OTUs in the GS library. Phylogenetic analysis showed that the sequences that belonged to the Firmicutes were the most predominant bacteria in both TS and GS libraries, representing 79.4% and 62.8% of the total clones, respectively. The remaining sequences belonged to Bacteroidetes, Proteobacteria, Actinobacteria, or were unclassified bacteria. Sequence analysis revealed that the TS and GS rumens harbored many novel sequences associated with uncultured bacteria that accounted for 63.6% and 46.8% of the total clones, respectively. Comparison of the composition and diversity of the TS and GS rumen bacteria revealed few overlapping known bacteria between the two breeds, and a higher diversity in TS. The rumen bacteria of the plateau sheep showed higher percentages of bacteria that belonged to Firmicutes and novel species compared with the low-elevation sheep. The unique bacterial community in the plateau sheep rumens is perhaps one of the major reasons that they can adapt to the harsh plateau environment. These results can help identify the rumen bacterial community of the ruminants in the QTP, and provide bacteria resources and basic data to improve ruminant productivity.

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.

Science.gov (United States)

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.

Effect of pH buffering capacity and sources of dietary sulfur on rumen fermentation, sulfide production, methane production, sulfate reducing bacteria, and total Archaea in in vitro rumen cultures.

Science.gov (United States)

Wu, Hao; Meng, Qingxiang; Yu, Zhongtang

2015-06-01

The effects of three types of dietary sulfur on in vitro fermentation characteristics, sulfide production, methane production, and microbial populations at two different buffer capacities were examined using in vitro rumen cultures. Addition of dry distilled grain with soluble (DDGS) generally decreased total gas production, degradation of dry matter and neutral detergent fiber, and concentration of total volatile fatty acids, while increasing ammonia concentration. High buffering capacity alleviated these adverse effects on fermentation. Increased sulfur content resulted in decreased methane emission, but total Archaea population was not changed significantly. The population of sulfate reducing bacteria was increased in a sulfur type-dependent manner. These results suggest that types of dietary sulfur and buffering capacity can affect rumen fermentation and sulfide production. Diet buffering capacity, and probably alkalinity, may be increased to alleviate some of the adverse effects associated with feeding DDGS at high levels. Copyright © 2015 Elsevier Ltd. All rights reserved.

Rumen escape protein in grass and grass silage deterimened with a nylon bag and an enzymatic technique

NARCIS (Netherlands)

Cone, J.W.; Gelder, van A.H.; Mathijssen-Kamman, A.A.; Hindle, V.A.

2004-01-01

Rumen escape protein (REP) was determined for six grasses and 16 grass silages using a nylon bag technique and an in vitro technique using a proteolytic enzyme preparation of Streptomyces griseus. In vitro, the samples were incubated for 0, 1, 6 and 24 h. The highest correlation observed between

The effect of lactic acid bacteria included as a probiotic or silage inoculant on in vitro rumen digestibility, total gas and methane production

NARCIS (Netherlands)

Ellis, J.L.; Bannink, A.; Hindrichsen, I.K.; Kinley, R.D.; Pellikaan, W.F.; Milora, N.L.; Dijkstra, J.

2016-01-01

Through alterations in silage and rumen fermentation, lactic acid bacteria (LAB) silage inoculants may affect OM digestibility and methane (CH4) emissions. In order to identify LAB that may have beneficial effects on CH4 emissions and/or OM digestibility in vivo, a series of in vitro gas production

Sampling methods for rumen microbial counts by Real-Time PCR techniques

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S. Puppo

2010-02-01

Full Text Available Fresh rumen samples were withdrawn from 4 cannulated buffalo females fed a fibrous diets in order to quantify bacteria concentration in the rumen by Real-Time PCR techniques. To obtain DNA of a good quality from whole rumen fluid, eight (M1-M8 different pre-filtration methods (cheese cloths, glass-fibre and nylon filter in combination with various centrifugation speeds (1000, 5000 and 14,000 rpm were tested. Genomic DNA extraction was performed either on fresh or frozen samples (-20°C. The quantitative bacteria analysis was realized according to Real-Time PCR procedure for Butyrivibrio fibrisolvens reported in literature. M5 resulted the best sampling procedure allowing to obtain a suitable genomic DNA. No differences were revealed between fresh and frozen samples.

Proteolytic and ACE-inhibitory activities of probiotic yogurt containing non-viable bacteria as affected by different levels of fat, inulin and starter culture

OpenAIRE

Shakerian, Mansour; Razavi, Seyed Hadi; Ziai, Seyed Ali; Khodaiyan, Faramarz; Yarmand, Mohammad Saeid; Moayedi, Ali

2013-01-01

In this study, the effects of fat (0.5 %, 3.2 % and 5.0 %), inulin (0.0 and 1.0 %) and starter culture (0.0 %, 0.5 %, 1.0 % and 1.5 %) on the angiotensin converting enzyme (ACE)-inhibitory activity of probiotic yogurt containing non-viable bacteria were assessed. Proteolytic activities of bacteria were also investigated. Yogurts were prepared either using a sole yogurt commercial culture including Streptococcus thermophilus and Lactobacillus delbrueckii subs. bulgaricus or bifidobacterium ani...

Biochanin A improves fiber fermentation by cellulolytic bacteria

Science.gov (United States)

The objective was to determine the effect of the isoflavone biochanin A (BCA) on rumen cellulolytic bacteria and consequent fermentative activity. When bovine microbial rumen cell suspensions (n = 3) were incubated (24 h, 39 °C) with ground hay, cellulolytic bacteria proliferated, short chain fatty...

Rumen microbial changes in cattle fed diets with or without salinomycin.

Science.gov (United States)

Olumeyan, D B; Nagaraja, T G; Miller, G W; Frey, R A; Boyer, J E

1986-02-01

Four rumen-fistulated steers, randomly assigned to two groups (control and salinomycin fed) were used to monitor the changes in rumen microbial populations and volatile fatty acids (VFA) concentrations associated with feeding salinomycin (0.22 mg X kg-1 X day-1). Steers were adapted to an alfalfa hay and grain (80:20) diet before supplementing the diet with salinomycin, and then the diet was changed to 50:50 and 20:80 ratios of alfalfa hay to grain at 2-week intervals. Rumen samples for total and selective enumeration of anaerobic bacteria. VFA analysis, and enumeration of protozoa were collected during the 80:20 alfalfa hay-to-grain diet before salinomycin feeding, and during the 80:20, 50:50, and 20:80 hay-to-grain diets with salinomycin. At each sampling period, rumen samples were collected at 3 h after feeding on three consecutive days. Salinomycin feeding had no effect on rumen pH and total VFA concentration. The acetate-to-propionate ratio was significantly lower in salinomycin-fed steers than in the control. The molar proportion of butyrate increased in both control and salinomycin-fed steers. Total anaerobic bacterial counts were lower in salinomycin-fed steers than in the control steers after 8 weeks of salinomycin feeding. Salinomycin-resistant bacteria increased from 7.6 to 15.6% in salinomycin-fed steers but remained unchanged in control steers. Salinomycin had no effect on cellulolytic and lactate-utilizing bacteria, but the proportion of amylolytic bacteria was higher in salinomycin-fed steers than in control steers. The total number of protozoa decreased initially in salinomycin-fed steers. The initial reduction was due to reduced numbers of Entodinium species. Holotrichs were unaffected by salinomycin feeding.

Temporal metagenomic and metabolomic characterisation of fresh perennial ryegrass degradation by rumen bacteria

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Olga Mayorga

2016-11-01

Full Text Available Understanding the relationship between ingested plant material and the attached microbiome is essential for developing methodologies to improve ruminant nutrient use efficiency. We have previously shown that perennial ryegrass (PRG rumen bacterial colonisation events follow a primary (up to 4 h and secondary (after 4 h pattern based on the differences in diversity of the attached bacteria. In this study we investigated temporal niche specialisation of primary and secondary populations of attached rumen microbiota using metagenomic shotgun sequencing as well as monitoring changes in the plant chemistry using mid-infrared spectroscopy (FT-IR. Metagenomic Rapid Annotation using Subsystem Technology (MG-RAST taxonomical analysis of shotgun metagenomic sequences showed that the genera Butyrivibrio, Clostridium, Eubacterium, Prevotella and Selenomonas dominated the attached microbiome irrespective of time. MG-RAST also showed that Acidaminococcus, Bacillus, Butyrivibrio and Prevotella rDNA increased in read abundance during secondary colonisation, whilst Blautia decreased in read abundance. MG-RAST Clusters of Orthologous Groups (COG functional analysis also showed that the primary function of the attached microbiome was categorised broadly within ‘metabolism’; predominantly amino acid, carbohydrate, and lipid metabolism and transport. Most sequence read abundances (51.6, 43.8, and 50.0% of COG families pertaining to amino acid, carbohydrate and lipid metabolism, respectively within these categories were higher in abundance during secondary colonisation. Kyoto encyclopedia of genes and genomes (KEGG pathways analysis confirmed that the PRG- attached microbiota present at 1 and 4 h of rumen incubation possess a similar functional capacity, with only a few pathways being uniquely found in only one incubation time point only. FT-IR data for the plant residues also showed that the main changes in plant chemistry between primary and secondary

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

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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.

Rumen microbial genomics

International Nuclear Information System (INIS)

Morrison, M.; Nelson, K.E.

2005-01-01

Improving microbial degradation of plant cell wall polysaccharides remains one of the highest priority goals for all livestock enterprises, including the cattle herds and draught animals of developing countries. The North American Consortium for Genomics of Fibrolytic Ruminal Bacteria was created to promote the sequencing and comparative analysis of rumen microbial genomes, offering the potential to fully assess the genetic potential in a functional and comparative fashion. It has been found that the Fibrobacter succinogenes genome encodes many more endoglucanases and cellodextrinases than previously isolated, and several new processive endoglucanases have been identified by genome and proteomic analysis of Ruminococcus albus, in addition to a variety of strategies for its adhesion to fibre. The ramifications of acquiring genome sequence data for rumen microorganisms are profound, including the potential to elucidate and overcome the biochemical, ecological or physiological processes that are rate limiting for ruminal fibre degradation. (author)

Effect of inclusion of different levels of silage on rumen microbial population and microbial protein synthesis in dairy steers fed on rice straw

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Thien Truong Giang Nguyen

2017-02-01

Full Text Available Objective Leucaena leucocephala (Leucaena is a perennial tropical legume that can be directly grazed or harvested and offered to ruminants as hay, silage, or fresh. However, Leucaena contain phenolic compounds, which are considered anti-nutritional factors as these may reduce intake, digestibility and thus animal performance. Therefore, the objective of this experiment was to determine effects of Leucaena silage (LS feeding levels on rumen microbial populations, N-balance and microbial protein synthesis in dairy steers. Methods Four, rumen fistulated dairy steers with initial weight of 167±12 kg were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. Treatments were as followings: T1 = untreated rice straw (RS; Control, T2 = 70% RS+30% LS, T3 = 40% RS+60% LS, and T4 = 100% LS. Dairy steers were fed rice straw and LS ad libitum and supplemented with concentrate at 0.2% of body weight/d. Results Results revealed that the rumen microbial population, especially cellulolytic, proteolytic bacteria and fungal zoospores were enhanced in steers that received 60% of LS (p0.05. Protozoal population was linearly decreased with increasing level of LS (p<0.05. Moreover, N-balance and microbial protein synthesis were enhanced by LS feeding (p<0.05 and were the highest in 60% LS group. Conclusion Based on this study, it could be concluded that replacement of RS with 60% LS significantly improved microbial population and microbial protein synthesis in diary steers.

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

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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.

Changes in the rumen bacterial community in response to sunflower oil and fish oil supplements in the diet of dairy sheep.

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Belenguer, A; Toral, P G; Frutos, P; Hervás, G

2010-07-01

Rumen microbial biohydrogenation of dietary unsaturated fatty acids has a major effect on the process of developing healthier dairy products. This study aimed to investigate in vivo the effect of diet supplementation with sunflower (SO) and fish (FO) oils on the rumen bacterial community in dairy sheep. First, 32 lactating ewes, divided in 8 lots of 4 animals each (2 lots per treatment), were fed a high-concentrate total mixed ration supplemented with 0, 2% SO, 1% FO, or 2% SO plus 1% FO. After 21 d, rumen fluid samples were taken from each lot for DNA extraction and fluorescence in situ hybridization (FISH) analysis. In a second experiment, 5 cannulated ewes were first fed the same TMR, with the exception of a higher forage level, and then changed to the same diet supplemented with 2% SO plus 1% FO. After 0, 3, and 10 d, rumen content samples were taken for DNA extraction and FISH analysis (fluid). Total bacteria and the Butyrivibrio group were studied in microbial DNA by terminal RFLP analysis (T-RFLP), and real-time PCR was used to quantify Butyrivibrio bacteria that produce vaccenic acid or stearic acid. In rumen fluid samples, total bacteria and clostridial clusters IX and XIV were analyzed by FISH. Dietary supplementation with SO plus FO seemed to induce important changes in the total bacteria and Butyrivibrio populations, and a high interindividual variation was observed, and the speed of the effect of the lipid supplementation depended on the individual microbial composition. Analysis by T-RFLP and FISH showed increases in cluster IX bacteria with SO plus FO supplementation, presumably Quinella-like microorganisms. The abundances of vaccenic acid- and stearic acid-producing Butyrivibrio relative to total bacteria, estimated by real time PCR, were low (0.28 and 0.18%, respectively, in rumen fluid, and 0.86 and 0.81% in rumen contents) and only that of SA-producing bacteria seemed to be reduced by diets containing FO, although differences were only

Rumen microbiota and dietary fat: a mutual shaping.

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Enjalbert, F; Combes, S; Zened, A; Meynadier, A

2017-10-01

Although fat content in usual ruminant diets is very low, fat supplements can be given to farm ruminants to modulate rumen activity or the fatty acid (FA) profile of meat and milk. Unsaturated FAs, which are dominant in common fat sources for ruminants, have negative effects on microbial growth, especially protozoa and fibrolytic bacteria. In turn, the rumen microbiota detoxifies unsaturated FAs (UFAs) through a biohydrogenation (BH) process, transforming dietary UFAs with cis geometrical double-bonds into mainly trans UFAs and, finally, into saturated FAs. Culture studies have provided a large amount of data regarding bacterial species and strains that are affected by UFAs or involved in lipolysis or BH, with a major focus on the Butyrivibrio genus. More recent data using molecular approaches to rumen microbiota extend and challenge these data, but further research will be necessary to improve our understanding of fat and rumen microbiota interactions. © 2017 The Society for Applied Microbiology.

Bacterial Population Adherent to the Epithelium on the Roo of the Dorsal Rumen of Sheep †

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Dehority, Burk A.; Grubb, Jean A.

1981-01-01

By anaerobic procedures, the total number of adherent bacteria was determined on tissue samples obtained from the roof of the dorsal rumen of three sheep. After four washings, 1.91 × 107, 0.34 × 107, and 1.23 × 107 bacteria per cm2 were still attached to the rumen epithelium in sheep 1, 2, and 3, respectively. A total of 95 strains of bacteria were isolated from these three samples. Based on morphology, Gram stain, anaerobiosis, motility, and fermentation end products, they were presumptively...

Metatranscriptome Sequencing Reveals Insights into the Gene Expression and Functional Potential of Rumen Wall Bacteria

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Evelyne Mann

2018-01-01

Full Text Available Microbiota of the rumen wall constitute an important niche of rumen microbial ecology and their composition has been elucidated in different ruminants during the last years. However, the knowledge about the function of rumen wall microbes is still limited. Rumen wall biopsies were taken from three fistulated dairy cows under a standard forage-based diet and after 4 weeks of high concentrate feeding inducing a subacute rumen acidosis (SARA. Extracted RNA was used for metatranscriptome sequencing using Illumina HiSeq sequencing technology. The gene expression of the rumen wall microbial community was analyzed by mapping 35 million sequences against the Kyoto Encyclopedia for Genes and Genomes (KEGG database and determining differentially expressed genes. A total of 1,607 functional features were assigned with high expression of genes involved in central metabolism, galactose, starch and sucrose metabolism. The glycogen phosphorylase (EC:2.4.1.1 which degrades (1->4-alpha-D-glucans was among the highest expressed genes being transcribed by 115 bacterial genera. Energy metabolism genes were also highly expressed, including the pyruvate orthophosphate dikinase (EC:2.7.9.1 involved in pyruvate metabolism, which was covered by 177 genera. Nitrogen metabolism genes, in particular glutamate dehydrogenase (EC:1.4.1.4, glutamine synthetase (EC:6.3.1.2 and glutamate synthase (EC:1.4.1.13, EC:1.4.1.14 were also found to be highly expressed and prove rumen wall microbiota to be actively involved in providing host-relevant metabolites for exchange across the rumen wall. In addition, we found all four urease subunits (EC:3.5.1.5 transcribed by members of the genera Flavobacterium, Corynebacterium, Helicobacter, Clostridium, and Bacillus, and the dissimilatory sulfate reductase (EC 1.8.99.5 dsrABC, which is responsible for the reduction of sulfite to sulfide. We also provide in situ evidence for cellulose and cellobiose degradation, a key step in fiber-rich feed

Effect of Propionibacterium acidipropionici P169 on the rumen and faecal microbiota of beef cattle fed a maize-based finishing diet.

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Azad, E; Narvaez, N; Derakhshani, H; Allazeh, A Y; Wang, Y; McAllister, T A; Khafipour, E

2017-10-13

Direct fed microbial supplementation with lactic acid utilising bacteria (i.e. Propionibacterium acidipropionici P169) has been shown to alleviate the severity of subacute ruminal acidosis in high-grain fed beef cattle. This study was carried out to explore the impact of P169 supplementation on modulating rumen and hindgut microbiota of high-grain fed steers. Seven ruminally-canulated high-grain fed steers were randomly assigned to two treatment groups: control diet (n=3) and the same diet supplemented with P169 added at a rate of 1×10 11 cfu/head/d (n=4). Samples were collected every 28 days for a 101 d period (5 time points) and subjected to qPCR quantification of P169 and high-throughput sequencing of bacterial V4 16S rRNA genes. Ruminal abundance of P169 was maintained at elevated levels (P=0.03) both in liquid and solid fractions post supplementation. Concomitant with decreased proportion of amylolytic (such as Prevotella) and key lactate-utilisers (such as Veillonellaceae and Megasphaera), the proportions of cellulolytic bacterial lineages (such as Ruminococcaceae, Lachnospiraceae, Clostridiaceae, and Christensenellaceae) were enriched in the rumen microbiota of P169-supplemented steers. These, coupled with elevated molar proportions of branched-chain fatty acids and increased concentration of ammonia in the rumen content of P169-supplemented steers, indicated an improved state of fibrolytic and proteolytic activity in response to P169 supplementation. Further, exploring the hindgut microbiota of P169-supplemented steers revealed enrichment of major amylolytic bacterial lineages, such as Prevotella, Blautia, and Succinivibrionaceae, which might be indicative of an increased availability of carbohydrates in the hindgut ecosystem following P169 supplementation. Collectively, the present study provides insights into the microbiota dynamics that underlie the P169-associated shifts in the rumen fermentation profile of high-grain fed steers.

Quantitative comparisons of select cultured and uncultured microbial populations in the rumen of cattle fed different diets

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Kim Minseok

2012-09-01

Full Text Available Abstract Background The number and diversity of uncultured ruminal bacterial and archaeal species revealed by 16S rRNA gene (rrs sequences greatly exceeds that of cultured bacteria and archaea. However, the significance of uncultured microbes remains undetermined. The objective of this study was to assess the numeric importance of select uncultured bacteria and cultured bacteria and the impact of diets and microenvironments within cow rumen in a comparative manner. Results Liquid and adherent fractions were obtained from the rumen of Jersey cattle fed hay alone and Holstein cattle fed hay plus grain. The populations of cultured and uncultured bacteria present in each fraction were quantified using specific real-time PCR assays. The population of total bacteria was similar between fractions or diets, while total archaea was numerically higher in the hay-fed Jersey cattle than in the hay-grain-fed Holstein cattle. The population of the genus Prevotella was about one log smaller than that of total bacteria. The populations of Fibrobacter succinogenes, Ruminococcus flavefaciens, the genus Butyrivibrio, and R. albus was at least one log smaller than that of genus Prevotella. Four of the six uncultured bacteria quantified were as abundant as F. succinogenes, R. flavefaciens and the genus Butyrivibrio. In addition, the populations of several uncultured bacteria were significantly higher in the adherent fractions than in the liquid fractions. These uncultured bacteria may be associated with fiber degradation. Conclusions Some uncultured bacteria are as abundant as those of major cultured bacteria in the rumen. Uncultured bacteria may have important contribution to ruminal fermentation. Population dynamic studies of uncultured bacteria in a comparative manner can help reveal their ecological features and importance to rumen functions.

Monensin and Nisin Affect Rumen Fermentation and Microbiota Differently In Vitro

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Junshi Shen

2017-06-01

Full Text Available Nisin, a bacteriocin, is a potential alternative to antibiotics to modulate rumen fermentation. However, little is known about its impacts on rumen microbes. This study evaluated the effects of nisin (1 and 5 μM on in vitro rumen fermentation characteristics, microbiota, and select groups of rumen microbes in comparison with monensin (5 μM, one of the most commonly used ionophores in ruminants. Nisin had greater effects than monensin in inhibiting methane production and decreasing acetate/propionate ratio. Unlike monensin, nisin had no adverse effect on dry matter digestibility. Real-time PCR analysis showed that both monensin and nisin reduced the populations of total bacteria, fungi, and methanogens, while the population of protozoa was reduced only by monensin. Principal component analysis of bacterial 16S rRNA gene amplicons showed a clear separation between the microbiota shaped by monensin and by nisin. Comparative analysis also revealed a significant difference in relative abundance of some bacteria in different taxa between monensin and nisin. The different effects of monensin and nisin on microbial populations and bacterial communities are probably responsible for the discrepancy in their effects on rumen fermentation. Nisin may have advantages over monensin in modulating ruminal microbial ecology and reducing ruminant methane production without adversely affecting feed digestion, and thus it may be used as a potential alternative to monensin fed to ruminants.

Ammonia production by ruminal microorganisms and enumeration, isolation, and characterization of bacteria capable of growth on peptides and amino acids from the sheep rumen.

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Eschenlauer, S C P; McKain, N; Walker, N D; McEwan, N R; Newbold, C J; Wallace, R J

2002-10-01

Excessive NH(3) production in the rumen is a major nutritional inefficiency in ruminant animals. Experiments were undertaken to compare the rates of NH(3) production from different substrates in ruminal fluid in vitro and to assess the role of asaccharolytic bacteria in NH(3) production. Ruminal fluid was taken from four rumen-fistulated sheep receiving a mixed hay-concentrate diet. The calculated rate of NH(3) production from Trypticase varied from 1.8 to 19.7 nmol mg of protein(-1) min(-1) depending on the substrate, its concentration, and the method used. Monensin (5 micro M) inhibited NH(3) production from proteins, peptides, and amino acids by an average of 28% with substrate at 2 mg/ml, compared to 48% with substrate at 20 mg/ml (P = 0.011). Of the total bacterial population, 1.4% grew on Trypticase alone, of which 93% was eliminated by 5 micro M monensin. Many fewer bacteria (0.002% of the total) grew on amino acids alone. Nineteen isolates capable of growth on Trypticase were obtained from four sheep. 16S ribosomal DNA and traditional identification methods indicated the bacteria fell into six groups. All were sensitive to monensin, and all except one group (group III, similar to Atopobium minutum), produced NH(3) at >250 nmol min(-1) mg of protein(-1), depending on the medium, as determined by a batch culture method. All isolates had exopeptidase activity, but only group III had an apparent dipeptidyl peptidase I activity. Groups I, II, and IV were most closely related to asaccharolytic ruminal and oral Clostridium and Eubacterium spp. Group V comprised one isolate, similar to Desulfomonas piger (formerly Desulfovibrio pigra). Group VI was 95% similar to Acidaminococcus fermentans. Growth of the Atopobium- and Desulfomonas-like isolates was enhanced by sugars, while growth of groups I, II, and V was significantly depressed by sugars. This study therefore demonstrates that different methodologies and different substrate concentrations provide an explanation

[Screening of harmine tolerance/degrading bacteria from camel rumen].

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An, Dengdi; Zhu, Yanlei; Tang, Jing; Ye, Yongxia; Zeng, Xianchun

2010-08-01

Peganum harmala, a famous traditional Chinese drug, contains a variety of alkaloids and toxic for many animals. Camels mainly live in desert or semi-desert areas, with the robust gastrointestine system in digesting various feed including toxic plants without disease symptoms. Camel rumen content was used as the inoculant to inoculate medium M98-5 which contains 100 mg x L(-1) harmin and cultivated for 5 days. Upto 5 subculturings, strains that could degrading or tolerant harmine were isolated. Their conversion activity was determined by thin-layer chromatography. The taxonomic position of the strains were identified based on 16S rRNA sequences analysis. 15 out of the 29 isolates have harmine degrading activity. Most of the isolates are identified as the members of the Genera Lactobacillus (16 strains, 55%), Shigella (7 strains, 24%) and Bacillus (4 strains, 13.8%). Only one strain belong to genus Enterococcus and one belong to genus Megasphaera. The results indicated that the harmine tolerance/degrading communities of camel rumen are limited and only Lactobacillus have harmine-degrading activity.

Methane emission and community composition patterns of rumen bacteria and methanogens in Holstein dairy cows as affected by silage type and dietary fat

DEFF Research Database (Denmark)

Poulsen, Morten; Højberg, Ole; Canibe, Nuria

) to investigate effects of silages with different methanogenic potential (early grass, late grass, and maize) combined with a CH4-reducing feed additive (crushed rapeseed) on bacterial and methanogenic communities in the rumen. Bacterial and methanogenic community patterns were evaluated by T-RFLP analysis of 16S...... rRNA and methyl co-enzyme M reductase (mcrA) genes, respectively. Methanogen abundances were evaluated by qPCR using two mcrA-targeting primer sets. Silage type significantly affected CH4 emissions and rumen acetate:propionate ratios, being highest for late grass and lowest for maize. Dietary fat...... significantly reduced the gross energy lost as CH4 regardless of silage type. Silage type significantly affected the bacterial community composition pattern; the grass silages favored potential hemicellulose- and cellulose-degrading bacteria, while the maize silage mainly favored potentially starch...

Synthesis of microbial nitrogen compounds in the rumen and their digestion in the small intestine

International Nuclear Information System (INIS)

Smith, R.H.; Salter, D.N.; McAllan, A.B.; Williams, A.P.

1976-01-01

Pathways and efficiency of NPN utilization were studied after introducing isonitrogenous feeds containing urea labelled with 15 N with or without groundnut protein into the rumen of the young steer. The 15 N abundance in rumen ammonia decreased from 1 h after the dose whereas that in suspended bacteria (only a small part of the total population but regarded, with reservation, as representative) increased to a peak value at 3-5 h. Patterns of incorporation into different nitrogen constituents of the bacteria indicated that part of the 15 N first moved rapidly into a labile pool, which included amide nitrogen, and then moved from this pool into other amino acids. Different amino acids were labelled to different extents (lowest for proline and arginine, highest for aspartic acid and alanine), presumably as different amounts of pre-formed amino acids were used in protein synthesis. Differences largely disappeared when the diet with urea as the only nitrogen source was given. Estimates of microbial 15 N recovery at the duodenum confirmed earlier findings that overall incorporation was least when the latter diet was given, and no improvement was seen when the urea was given in three doses over 4 h instead of in a single dose. Lactosyl, like glucosyl, ureide was degraded fairly slowly in the rumen and is being studied as a possible NPN source likely to yield simultaneous ammonia and energy for microbial growth. Studies on the origin and composition of nitrogen compounds entering the duodenum showed that the RNA of rumen bacteria (labelled with 32 P) contributed 82-109% of the RNA entering the duodenum of the steer. It was also shown that protecting dietary casein with formaldehyde reduced its degradability in the rumen to such an extent that a deficiency of fermentable nitrogen in that organ ensued. In other experiments with two calves receiving flaked maize and hay, net digestibility of nitrogen compounds between duodenum and ileum was about 60%. True digestibility

Isolation and characterisation of obligately anaerobic, lipolytic bacteria from the rumen of red deer.

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Jarvis, G N; Strömpl, C; Moore, E R; Thiele, J H

1998-03-01

Two Gram-positive, obligately anaerobic, lipolytic bacteria, isolates LIP4 and LIP5, were obtained from the rumen contents of juvenile red deer. These mesophilic bacterial strains were capable of hydrolysing the neutral lipids, tallow, tripalmitin and oliver oil, into their constituent free long-chain fatty acid and glycerol moieties. The latter compound was dissimilated by both isolates, with isolate LIP4 producing propionate as the predominant product, while isolate LIP5 produced acetate, ethanol and succinate. The lactate-utilising isolate LIP4 grew on a limited range of saccharide substrates including glucose, fructose and ribose, and exhibited an unusual cell wall structure and morphology. The isolate LIP5 grew upon a wider range of saccharides, but was unable to use lactate as a substrate. Based upon phenotypic and 16S rRNA gene sequence analyses, isolate LIP4 clusters with species in the genus Propionibacterium, while isolate LIP5 is a member of clostridial cluster XIVa.

Replacement of alfalfa hay (Medicago sativa L.) with subabul (Leucaena leucocephala) leaf meal in diets of Najdi goats: effect on digestion activity of rumen microorganisms.

Science.gov (United States)

Mohammadabadi, Tahereh; Jolazadeh, Alireza

2017-08-01

This study investigated the effect of replacing alfalfa hay by subabul leaf meal (SLM) on digestion, fermentation parameters and rumen bacteria and fungi activity of Najdi goats. Six Najdi goats (150 ± 15 days of age and initial body weight of 35 ± 1.1 kg) were randomly assigned to one of two dietary treatments in a balanced completely randomized design (three goats per treatment) for 56 days. Experimental treatments included alfalfa hay as control group and diet containing SLM (SLM replacing alfalfa hay at 50% level). Bacterial and fungi activity and rumen fermentation parameters of animals fed experimental diets were determined. Dry matter disappearance (DMD) was unaffected by replacing SLM with alfalfa hay for both rumen bacteria and fungi in different incubation times, except for 48 h of incubation in specific culture medium of mixed rumen bacteria, which decreased for SLM group (P > 0.05). NDF disappearance (NDFD) and ADF disappearance (ADFD) after 24 and 48 h of incubation in specific culture medium of mixed rumen bacteria was not affected by experimental diets (P > 0.05). However, 72 h after incubation, NDFD in SLM treatment decreased (P > 0.05). Gas production parameters of rumen bacteria were similar for both experimental diets, but partitioning factor (PF), efficiency microbial biomass production (EMBP), microbial protein production (MP), and organic matter truly digested (OMTD) decreased (p alfalfa hay by SLM had no major effect on rumen microorganisms' activity of Najdi goats, so it may be used as an alternative for alfalfa (at 50% level) in susceptible areas.

Molecular diversity of rumen bacterial communities from tannin-rich and fiber-rich forage fed domestic Sika deer (Cervus nippon) in China.

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Li, Zhi Peng; Liu, Han Lu; Li, Guang Yu; Bao, Kun; Wang, Kai Ying; Xu, Chao; Yang, Yi Feng; Yang, Fu He; Wright, André-Denis G

2013-07-08

Sika deer (Cervus nippon) have different dietary preferences to other ruminants and are tolerant to tannin-rich plants. Because the rumen bacteria in domestic Sika deer have not been comprehensively studied, it is important to investigate its rumen bacterial population in order to understand its gut health and to improve the productivity of domestic Sika deer. The rumen bacterial diversity in domestic Sika deer (Cervus nippon) fed oak leaves- (OL group) and corn stalks-based diets (CS group) were elucidated using 16S rRNA gene libraries and denaturing gradient gel electrophoresis (DGGE). Overall, 239 sequences were examined from the two groups, 139 clones from the OL group were assigned to 57 operational taxonomic units (OTUs) and 100 sequences from the CS group were divided into 50 OTUs. Prevotella-like sequences belonging to the phylum Bacteroidetes were the dominant bacteria in both groups (97.2% OL and 77% CS), and sequences related to Prevotella brevis were present in both groups. However, Prevotella shahii-like, Prevotella veroralis-like, Prevotella albensis-like, and Prevotella salivae-like sequences were abundant in the OL group compared to those in the CS group, while Succinivibrio dextrinosolvens-like and Prevotella ruminicola-like sequences were prevalent in the CS group. PCR-DGGE showed that bacterial communities clustered with respect to diets and the genus Prevotella was the dominant bacteria in the rumen of domestic Sika deer. However, the distribution of genus Prevotella from two groups was apparent. In addition, other fibrolytic bacteria, such as Clostridium populeti and Eubacterium cellulosolvens were found in the rumen of domestic Sika deer. The rumen of domestic Sika deer harbored unique bacteria which may represent novel species. The bacterial composition appeared to be affected by diet, and sequences related to Prevotella spp. may represent new species that may be related to the degradation of fiber biomass or tannins. Moreover, the mechanism

Response of the Rumen Microbiota of Sika Deer (Cervus nippon Fed Different Concentrations of Tannin Rich Plants.

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Zhipeng Li

Full Text Available High throughput sequencing was used to examine the rumen microbiota of sika deer fed high (OLH and low concentration (OLL of tannin rich oak leaves. The results showed that Prevotella spp. were the most dominant bacteria. The most predominant methanogens were the members of the order Methanoplasmatales. The dominant rumen protozoa were Entodinium longinucleatum, Eudiplodinium maggii, and Epidinium caudatum, and the fungal communities were mostly represented by Piromyces spp. Moreover, the relative abundance of Pseudobutyrivibrio spp. (P=0.026, unidentified bacteria (P=0.028, and Prevotella spp. (P=0.022 was lower in the OLH group than in the OLL group. The concentration of propionate in the OLH group was greater than in the OLL group (P=0.006. Patterns of relationships showed that methanogens belonging to the order Methanoplasmatales were negatively correlated with Treponema spp., Ent. Longinucleatum, and acetate. Methanosphaera stadtmanae was positively correlated to propionate, while Methanobrevibacter ruminantium was negatively associated with Methanobrevibacter thaueri and Methanobrevibacter millerae. Tannins altered the rumen microbes and fermentation patterns. However, the response of the entire rumen microbiota and the relationship between rumen microorganisms and the fermentation parameters were not fully understood.

Rumen derived anaerobic digestion of water hyacinth (Eicchornia ...

African Journals Online (AJOL)

STORAGESEVER

2009-09-01

Sep 1, 2009 ... The agar plates were then incubated anaerobically at 37°C for 24 h. The digesters were seeded with rumen bacteria and immersed into water bath operated at 37°C. During the anaerobic digestion, volume of biogas produced was recorded accordingly. This paper, therefore, suggests ways by which water.

Isolation and characterization of novel lipases/esterases from a bovine rumen metagenome.

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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.

Rumen derived anaerobic digestion of water hyacinth (Eicchornia ...

African Journals Online (AJOL)

The agar plates were then incubated anaerobically at 37°C for 24 h. The digesters were seeded with rumen bacteria and immersed into water bath operated at 37°C. During the anaerobic digestion, volume of biogas produced was recorded accordingly. This paper, therefore, suggests ways by which water hyacinth can be ...

Exploring the bovine rumen bacterial community from birth to adulthood.

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Jami, Elie; Israel, Adi; Kotser, Assaf; Mizrahi, Itzhak

2013-06-01

The mammalian gut microbiota is essential in shaping many of its host's functional attributes. One such microbiota resides in the bovine digestive tract in a compartment termed as the rumen. The rumen microbiota is necessary for the proper physiological development of the rumen and for the animal's ability to digest and convert plant mass into food products, making it highly significant to humans. The establishment of this microbial population and the changes occurring with the host's age are important for understanding this key microbial community. Despite its importance, little information about colonization of the microbial populations in newborn animals, and the gradual changes occurring thereafter, exists. Here, we characterized the overall bovine ruminal bacterial populations of five age groups, from 1-day-old calves to 2-year-old cows. We describe the changes occurring in the rumen ecosystem after birth, reflected by a decline in aerobic and facultative anaerobic taxa and an increase in anaerobic ones. Some rumen bacteria that are essential for mature rumen function could be detected as early as 1 day after birth, long before the rumen is active or even before ingestion of plant material occurs. The diversity and within-group similarity increased with age, suggesting a more diverse but homogeneous and specific mature community, compared with the more heterogeneous and less diverse primary community. In addition, a convergence toward a mature bacterial arrangement with age was observed. These findings have also been reported for human gut microbiota, suggesting that similar forces drive the establishment of gut microbiotas in these two distinct mammalian digestive systems.

16S/18S ribosomal DNA clone library analysis of rumen microbial diversity

International Nuclear Information System (INIS)

Wright, A.G.; Kiyoshi Tajima; Aminov, R.I.

2005-01-01

The rumen contains a complex ecosystem where billions of bacteria, archaea, protozoa and fungi reside. This diverse microbiota is well adapted to live in the rumen and play an important role in the digestion of feed and nutrient supply to the host in the form of microbial protein and volatile fatty acids. It is estimated that the rumen microbial population consists of about 10 6 protozoa/ml, 10 3 -10 7 fungi/ml, 10 10 bacteria/ml, and 10 9 methanogens/ml. To better understand the complex relationships in the rumen, it is necessary to gain an insight into the diversity of the rumen microbes and how the quantity and composition of rumen micro-organisms are altered by a number of different host factors such as age, genetics and diet. In the past, the diversity of micro-organisms from the digestive tracts of domesticated ruminants has been identified by classical microbiological techniques. However, given the fastidious growth requirements of rumen micro-organisms, it is reasonable to concede that the culture-dependent methods may select against some species, or taxonomic groups, leading researchers to underestimate the microbial diversity that is actually present in the rumen. In fact, it has been speculated that 90% of micro-organisms in nature have escaped traditional cultivation methods. Therefore, a major challenge in microbial ecology has been to assess the diversity and structure of natural microbial communities. The field of molecular biology has advanced with many innovative technological breakthroughs. The ability to extract and to isolate high-molecular weight DNA from rumen digesta, PCR amplify genes from specific microbial groups and obtain gene sequence data is now a routine event. The small subunit ribosomal RNA (SSU-rRNA) gene, called 16S in prokaryotes and 18S in eukaryotes, is the most widely used molecular marker to presumptively identify morphologically indistinguishable species, to infer their phylogenetic relationships, and to elucidate microbial

Comparison of Rumen and Manure Microbiomes and Implications for the Inoculation of Anaerobic Digesters.

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Ozbayram, Emine Gozde; Ince, Orhan; Ince, Bahar; Harms, Hauke; Kleinsteuber, Sabine

2018-02-14

Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methanogenic communities of rumen and manure samples were examined by 454 amplicon sequencing of bacterial 16S rRNA genes and mcrA genes, respectively. Rumen fluid samples were dominated by Prevotellaceae (29%), whereas Ruminococcaceae was the most abundant family in the manure samples (31%). Fibrobacteraceae (12%) and Bacteroidaceae (13%) were the second most abundant families in rumen fluid and manure, respectively. The high abundances of fiber-degrading bacteria belonging to Prevotellaceae and Fibrobacteraceae might explain the better performance of anaerobic digesters inoculated with rumen fluid. Members of the genus Methanobrevibacter were the predominant methanogens in the rumen fluid, whereas methanogenic communities of the manure samples were dominated by the candidate genus Methanoplasma . Our results suggest that inoculation or bioaugmentation with fiber-digesting rumen microbiota can enhance the anaerobic digestion of lignocellulosic biomass.

Comparison of Rumen and Manure Microbiomes and Implications for the Inoculation of Anaerobic Digesters

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Emine Gozde Ozbayram

2018-02-01

Full Text Available Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methanogenic communities of rumen and manure samples were examined by 454 amplicon sequencing of bacterial 16S rRNA genes and mcrA genes, respectively. Rumen fluid samples were dominated by Prevotellaceae (29%, whereas Ruminococcaceae was the most abundant family in the manure samples (31%. Fibrobacteraceae (12% and Bacteroidaceae (13% were the second most abundant families in rumen fluid and manure, respectively. The high abundances of fiber-degrading bacteria belonging to Prevotellaceae and Fibrobacteraceae might explain the better performance of anaerobic digesters inoculated with rumen fluid. Members of the genus Methanobrevibacter were the predominant methanogens in the rumen fluid, whereas methanogenic communities of the manure samples were dominated by the candidate genus Methanoplasma. Our results suggest that inoculation or bioaugmentation with fiber-digesting rumen microbiota can enhance the anaerobic digestion of lignocellulosic biomass.

Proteolytic activation transforms heparin cofactor II into a host defense molecule.

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Kalle, Martina; Papareddy, Praveen; Kasetty, Gopinath; Tollefsen, Douglas M; Malmsten, Martin; Mörgelin, Matthias; Schmidtchen, Artur

2013-06-15

The abundant serine proteinase inhibitor heparin cofactor II (HCII) has been proposed to inhibit extravascular thrombin. However, the exact physiological role of this plasma protein remains enigmatic. In this study, we demonstrate a previously unknown role for HCII in host defense. Proteolytic cleavage of the molecule induced a conformational change, thereby inducing endotoxin-binding and antimicrobial properties. Analyses employing representative peptide epitopes mapped these effects to helices A and D. Mice deficient in HCII showed increased susceptibility to invasive infection by Pseudomonas aeruginosa, along with a significantly increased cytokine response. Correspondingly, decreased levels of HCII were observed in wild-type animals challenged with bacteria or endotoxin. In humans, proteolytically cleaved HCII forms were detected during wounding and in association with bacteria. Thus, the protease-induced uncovering of cryptic epitopes in HCII, which transforms the molecule into a host defense factor, represents a previously unknown regulatory mechanism in HCII biology and innate immunity.

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

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Litai Zhang

Full Text Available 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.

Persistence of bacterial proteolytic enzymes in lake ecosystems.

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Kiersztyn, Bartosz; Siuda, Waldemar; Chróst, Ryszard J

2012-04-01

This study analyzes proteolytic enzyme persistence and the role of dead (or metabolically inactive) aquatic bacteria in organic matter cycling. Samples from four lakes of different trophic status were used. Irrespective of the trophic status of the examined lakes, bacterial aminopeptidases remained active even 72 h after the death of the bacteria that produced them. The total pool of proteolytic enzymes in natural lake water samples was also stable. We found that the rates of amino acid enzymatic release from proteinaceous matter added to preserved lake water sample were constant for at least 96 h (r(2)  = 0.99, n = 17, P ≤ 0.0001, V(max)  = 84.6 nM h(-1) ). We also observed that proteases built into bacterial cell debris fragments remained active for a long time, even after the total destruction of cells. Moreover, during 24 h of incubation time, about 20% of these enzymatically active fragments adsorbed onto natural seston particles, becoming a part of the 'attached enzymes system' that is regarded as the 'hot-spot' of protein degradation in aquatic ecosystems. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

The production of anaerobic bacteria and biogas from dairy cattle waste in various growth mediums

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Hidayati, Y. A.; Kurnani, T. B. A.; Marlina, E. T.; Rahmah, K. N.; Harlia, E.; Joni, I. M.

2018-02-01

The growth of anaerobic bacteria except the ruminal fluid quailty is strongly influenced by the media formulations. Previous researchers have set a standard media formulation for anaerobic bacteria from rumen, however the use of standard media formulations require chemicals with high cost. Moreover, other constraint of using standard media formulations is requires large quantities of media for anaerobic bacteria to grow. Therefore, it is necessary to find media with a new culture media formulation. Media used in this research were minimalist media consist of Nutrient Agar (NA), Lactose broth and rumen fluid; enriched media Rumen Fluid-Glucose-Agar (RGCA); and enriched media 98-5. The dairy cattle waste is utilized as source of anaerobic bacteria. The obtained data was analyzed by descriptive approach. The results showed that minimalist media produced anaerobic bacteria 2148 × 104 cfu/ml and biogas production: 1.06% CH4, 9.893% CO2; enriched media Rumen Fluid-Glucose-Agar (RGCA) produced anaerobic bacteria 1848 × 104 cfu/ml and biogas production 4.644% CH4, 9.5356% CO2; enriched media 98-5 produced anaerobic bacteria growth 15400 × 104 cfu/ml and biogas production 0.83% of CH4, 42.2% of CO2. It is conclude that the minimalist media was showed the best performance for the dairy cattle waste as source of anaerobic bacteria.

Enhancing the Resolution of Rumen Microbial Classification from Metatranscriptomic Data Using Kraken and Mothur

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Andre L. A. Neves

2017-12-01

Full Text Available The advent of next generation sequencing and bioinformatics tools have greatly advanced our knowledge about the phylogenetic diversity and ecological role of microbes inhabiting the mammalian gut. However, there is a lack of information on the evaluation of these computational tools in the context of the rumen microbiome as these programs have mostly been benchmarked on real or simulated datasets generated from human studies. In this study, we compared the outcomes of two methods, Kraken (mRNA based and a pipeline developed in-house based on Mothur (16S rRNA based, to assess the taxonomic profiles (bacteria and archaea of rumen microbial communities using total RNA sequencing of rumen fluid collected from 12 cattle with differing feed conversion ratios (FCR. Both approaches revealed a similar phyla distribution of the most abundant taxa, with Bacteroidetes, Firmicutes, and Proteobacteria accounting for approximately 80% of total bacterial abundance. For bacterial taxa, although 69 genera were commonly detected by both methods, an additional 159 genera were exclusively identified by Kraken. Kraken detected 423 species, while Mothur was not able to assign bacterial sequences to the species level. For archaea, both methods generated similar results only for the abundance of Methanomassiliicoccaceae (previously referred as RCC, which comprised more than 65% of the total archaeal families. Taxon R4-41B was exclusively identified by Mothur in the rumen of feed efficient bulls, whereas Kraken uniquely identified Methanococcaceae in inefficient bulls. Although Kraken enhanced the microbial classification at the species level, identification of bacteria or archaea in the rumen is limited due to a lack of reference genomes for the rumen microbiome. The findings from this study suggest that the development of the combined pipelines using Mothur and Kraken is needed for a more inclusive and representative classification of microbiomes.

Proteolytic activity and cooperative hemolytic effect of dermatophytes with different species of bacteria

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Keyvan Pakshir

2016-12-01

Conclusion: This study indicated that hemolysin and proteolytic enzymes potentially play a role in dermatophyte pathogenesis and S. aureus could be considered as a main bacterium for creation of co-hemolytic effect in association with dermatophyte species.

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

Science.gov (United States)

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

Isolation and characterization of a new hydrogen-utilizing bacterium from the rumen.

Science.gov (United States)

Rieu-Lesme, F; Fonty, G; Doré, J

1995-01-01

A new H2/CO2-utilizing acetogenic bacterium was isolated from the rumen of a mature deer. This is the first report of a spore-forming Gram-negative bacterial species from the rumen. The organism was a strictly anaerobic, motile rod and was able to grow autotrophically on hydrogen and carbon dioxide. Acetate was the major product detected. Glucose, fructose and lactate were also fermented heterotrophically. The optimum pH for growth was 7.0-7.5, and the optimum temperature was 37-42 degrees C. Yeast extract was required for growth and rumen fluid was highly stimulatory. The DNA base ratio was 52.9 +/- 0.5 mol% G+C. On the basis of these characteristics and fermentation products, the isolate was considered to be different from acetogenic bacteria described previously.

Fermentation of Rice Straw Uses Mix Inoculum of Anaerobe Facultative Bacteria Isolate from Buffalo Rumen

International Nuclear Information System (INIS)

Sasongko, W. T.; Irawan Sugoro

2004-01-01

Rice straw quality could be increased as feed by fermentation which has been mixed with bacteria inoculum from buffalo rumen. This experiment used rice straw from Atomita 4, four treatments and one control, i.e. A (rice straw, molasses 5 %, urea 5 %, and inoculum 10 %), B (rice straw, molasses 5 %, and urea 5 %), C (rice straw, molasses 5 %, and inoculum 10 %), D (rice straw and molasses 5 %), and K (control) have been used in this experiment. The parameters were digestibility of dry matter and organic matter, VFA, ammonia and in vitro gas production. The result, showed that the highest gas production, dry matter and organic matter digestibility occurred on A i.e. 17.48 ml/200 mg, 57.78%, and 52.39 %. The highest ammonia occurred on D (32.99 mg/100 ml) and the highest VFA occurred on C (12.36 mmol/100 ml). The concentration of ammonia and VFA of A significant to treatment of D and C). It may be concluded that the A treatment is the best and have potency to be develop. (author)

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

Science.gov (United States)

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. © Federation of European Microbiological Society 2014.

The complete genome sequence of Eubacterium limosum SA11, a metabolically versatile rumen acetogen

OpenAIRE

Kelly, William J.; Henderson, Gemma; Pacheco, Diana M.; Li, Dong; Reilly, Kerri; Naylor, Graham E.; Janssen, Peter H.; Attwood, Graeme T.; Altermann, Eric; Leahy, Sinead C.

2016-01-01

Acetogens are a specialized group of anaerobic bacteria able to produce acetate from CO2 and H2 via the Wood?Ljungdahl pathway. In some gut environments acetogens can compete with methanogens for H2, and as a result rumen acetogens are of interest in the development of microbial approaches for methane mitigation. The acetogen Eubacterium limosum SA11 was isolated from the rumen of a New Zealand sheep and its genome has been sequenced to examine its potential application in methane mitigation ...

Ruminal tryptophan-utilizing bacteria degrade ergovaline from tall fescue seed extract.

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Harlow, B E; Goodman, J P; Lynn, B C; Flythe, M D; Ji, H; Aiken, G E

2017-02-01

The objectives of this study were to evaluate degradation of ergovaline in a tall fescue [ (Schreb.) Darbysh.] seed extract by rumen microbiota ex vivo and to identify specific bacteria capable of ergovaline degradation in vitro. Rumen cell suspensions were prepared by harvesting rumen fluid from fistulated wether goats ( = 3), straining, and differential centrifugation. Suspensions were dispensed into anaerobic tubes with added Trypticase with or without extract (∼10 μg kg ergovaline). Suspensions were incubated for 48 h at 39°C. Samples were collected at 0, 24, and 48 h for ergovaline analysis and enumeration of hyper-ammonia producing (HAB) and tryptophan-utilizing bacteria. Ergovaline values were analyzed by repeated measures using the mixed procedure of SAS. Enumeration data were log transformed for statistical analysis. When suspensions were incubated with extract, 11 to 15% of ergovaline disappearance was observed over 48 h ( = 0.02). After 24 h, suspensions with added extract had 10-fold less HAB than controls ( = 0.04), but treatments were similar by 48 h ( = 1.00). However, after 24 h and 48 h, suspensions with extract had 10-fold more tryptophan-utilizing bacteria ( rumen pure cultures ( JB1, B159, HD4, B, F, MD1, SR) were evaluated for the ability to degrade ergovaline in vitro. Pure culture cell suspensions were incubated as described above and samples were taken at 0 and 48 h for ergovaline analysis. Data were analyzed using the ANOVA procedure of SAS. All HAB, including the isolates, tested degraded ergovaline (54 to 75%; bacteria tested did not degrade ergovaline. The results of this study indicate which rumen bacteria may play an important role in ergovaline degradation and that microbiological strategies for controlling their activity could have ramifications for fescue toxicosis and other forms of ergotism in ruminants.

Determination of rumen microbial growth in vitro form 32P-labelled phosphate incorporation

International Nuclear Information System (INIS)

Nevel, C.J. Van; Demeyer, D.I.

1977-01-01

The extracellular phosphate pool in incubations of rumen fluid or washed cell suspensions of mixed rumen bacteria (WCS) was labelled with 32 P. From the constant extracellular phosphate pool specific activity and the amount of radioactivity incorporated during incubation, the amount of P incorporated in the microbial fraction was calculated. From the value for nitrogen: P determined in microbial matter, the amount of N incorporated was calculated as a measure of microbial growth. Incorporation of soluble non-protein-N in incubations devoid of substrate protein was 50 and 80% of the values obtained using isotope method for rumen fluid and WCS respectively. Incorporation of 32 P in P-containing microbial components (mainly nucleic acids) was compared with net synthesis of these components in incubations of WCS. When N incorporation, calculated from results obtained using isotope method in incubations with rumen fluid, was compared with the amount of carbohydrate substrate fermented and the type of fermentation, values between 18.3 and 44.6 g N incorporated kg of organic matter fermented were obtained. The use of isotopes for determination of rumen microbial growth in vitro is critically discussed. (author)

Study on Protease Enzyme Produced by Pathogenic Bacteria

International Nuclear Information System (INIS)

Abd Elhakim, E.E.

2014-01-01

A total of 110 surface swabs were taken from skin of burned and post burn wounded patients. Infected swabs from burned patients were found to be 62.8% and 63.6%,while post burn wounded patients were 66.7% and 69.2% in male and female patients, respectively. The burned and post burn wounded patients were infected with bacteria in the order of gram negative bacilli >gram positive cocci>gram positive bacilli .Pseudomonas aeruginosa,Proteus mirabilis,Bacillus megaterium and Bacillus cereus showed the highest proteolytic activity. The effect of cultivation condition,different media, incubation time, incubation temperature, ph, metal ions, inhibitors and different doses of gamma radiation on the proteolytic activity of the tested bacterial isolates showing the highest proteolytic activity were studied. The tested bacterial isolates showed susceptibility to some antibiotics and showed resistance to the others. The effects of gamma radiation on the tested bacteria and on the susceptibility of the tested bacteria isolates to antibiotics under test were studied.

Optimal Cultivation Time for Yeast and Lactic Acid Bacteria in Fermented Milk and Effects of Fermented Soybean Meal on Rumen Degradability Using Nylon Bag Technique

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S. Polyorach

2016-09-01

Full Text Available The objectives of this study were to determine an optimal cultivation time for populations of yeast and lactic acid bacteria (LAB co-cultured in fermented milk and effects of soybean meal fermented milk (SBMFM supplementation on rumen degradability in beef cattle using nylon bag technique. The study on an optimal cultivation time for yeast and LAB growth in fermented milk was determined at 0, 4, 8, 24, 48, 72, and 96 h post-cultivation. After fermenting for 4 days, an optimal cultivation time of yeast and LAB in fermented milk was selected and used for making the SBMFM product to study nylon bag technique. Two ruminal fistulated beef cattle (410±10 kg were used to study on the effect of SBMFM supplementation (0%, 3%, and 5% of total concentrate substrate on rumen degradability using in situ method at incubation times of 0, 2, 4, 6, 12, 24, 48, and 72 h according to a Completely randomized design. The results revealed that the highest yeast and LAB population culture in fermented milk was found at 72 h-post cultivation. From in situ study, the soluble fractions at time zero (a, potential degradability (a+b and effective degradability of dry matter (EDDM linearly (p<0.01 increased with the increasing supplemental levels and the highest was in the 5% SBMFM supplemented group. However, there was no effect of SBMFM supplement on insoluble degradability fractions (b and rate of degradation (c. In conclusion, the optimal fermented time for fermented milk with yeast and LAB was at 72 h-post cultivation and supplementation of SBMFM at 5% of total concentrate substrate could improve rumen degradability of beef cattle. However, further research on effect of SBMFM on rumen ecology and production performance in meat and milk should be conducted using in vivo both digestion and feeding trials.

Degradation of Dehydrodivanillin by Anaerobic Bacteria from Cow Rumen Fluid

OpenAIRE

Chen, Wei; Ohmiya, Kunio; Shimizu, Shoichi; Kawakami, Hidekuni

1985-01-01

Dehydrodivanillin (DDV; 0.15 g/liter) was biodegradable at 37°C under strictly anaerobic conditions by microflora from cow rumen fluid to the extent of 25% within 2 days in a yeast extract medium. The anaerobes were acclimated on DDV for 2 weeks, leading to DDV-degrading microflora with rates of degradation eight times higher than those initially. Dehydrodivanillic acid and vanillic acid were detected in an ethylacetate extract of a DDV-enriched culture broth by thin-layer, gas, and high-perf...

Seasonal changes in the digesta-adherent rumen bacterial communities of dairy cattle grazing pasture

DEFF Research Database (Denmark)

Noel, Samantha Joan; Attwood, G T; Rakonjac, J

2017-01-01

The complex microbiota that resides within the rumen is responsible for the break-down of plant fibre. The bacteria that attach to ingested plant matter within the rumen are thought to be responsible for initial fibre degradation. Most studies examining the ecology of this important microbiome only.......1%), followed by Bacteroidetes (11.8%). This community differed between the seasons, returning to close to that observed in the same season one year later. These seasonal differences were only small, but were statistically significant (p diet...

Cell growth and proteolytic activity of Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii ssp. bulgaricus, and Streptococcus thermophilus in milk as affected by supplementation with peptide fractions.

Science.gov (United States)

Gandhi, Akanksha; Shah, Nagendra P

2014-12-01

The present investigation examined the effects of supplementation of milk peptide fractions produced by enzymatic hydrolysis on the fermentation of reconstituted skim milk (RSM). Changes in pH, cell growth, proteolytic activity, and angiotensin-converting enzyme (ACE)-inhibitory activity were monitored during fermentation of RSM by pure cultures of Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii ssp. bulgaricus, and Streptococcus thermophilus. The study showed that supplementation with peptide fractions of different molecular weights did not significantly affect the bacterial growth in RSM. All bacteria showed an increased proteolytic activity in RSM supplemented with large peptides (>10 kDa), and L. helveticus in general exhibited the highest proteolytic activity among the bacteria studied. The ACE-inhibitory activity was observed to be the maximum in RSM supplemented with larger peptides (>10 kDa) for all bacteria. The results suggest that proteolysis by bacteria leads to increased production of ACE-inhibitory peptides compared to the supplemented peptides produced by enzymatic hydrolysis.

Analysis of the rumen bacterial diversity of goats during shift from forage to concentrate diet

Czech Academy of Sciences Publication Activity Database

Grilli, D. J.; Fliegerová, Kateřina; Kopečný, Jan; Lama, S. P.; Egea, V.; Sohaefer, N.; Pereyra, C.; Ruiz, M. J.; Sosa, M. A.; Arenas, G. N.; Mrázek, Jakub

2016-01-01

Roč. 42, č. 1 (2016), s. 17-26 ISSN 1075-9964 Institutional support: RVO:67985904 Keywords : creole goats * rumen * bacteria Subject RIV: EE - Microbiology, Virology Impact factor: 2.278, year: 2016

Effect of tropical browse leaves supplementation on rumen enzymes of sheep and goats fed Dichanthium annulatum grass-based diets.

Science.gov (United States)

Singh, Sultan; Kundu, S S

2010-08-01

In a switch-over experiment, eight male animals, four each of sheep and goats of local breeds with mean body weight of 26. 8 +/- 2.0 and 30.0 +/- 2.1 kg, were fed Dichanthium annulatum (DA) grass and four browse species viz. Helictris isora, Securengia virosa, Leucaena leucocephala (LL) and Hardwickia binnata (HB) in four feeding trials to assess their supplementary effect on activity of rumen enzymes. The sheep and goats were offered DA grass with individual browse in 75:25 and 50:50 proportions, respectively, for more than 3 months during each feeding trial, and rumen liquor samples were collected twice at 0 and 4 h post feeding after 60 and 90 days of feeding. Glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and glutamate dehydrogenase (GDH) enzymes were determined in the bacteria and protozoa fractions of rumen liquor, while cellulase enzyme activity was measured in mixed rumen liquor. LL and HB had the highest and lowest contents of CP, while fibre contents were lower in early than later browse leaves. Supplementation of browse leaves significantly (P goats on all DA grass-browse-supplemented diets except DA-HB (42.8 units/mg protein), where activity was significantly (P Goat exhibited higher activities of GOT and GPT than sheep in both bacteria and protozoa fraction of rumen liquor, while cellulase activity was similar between the animal species on the grass-browse leaves diets. Results indicate that browse leaves supplementation affect the enzyme activities of sheep and goats rumen, while the goats rumen liquor had higher activities of GOT, GPT and GDH enzyme than sheep.

Effects of a specific blend of essential oils on apparent nutrient digestion, rumen fermentation and rumen microbial populations in sheep fed a 50:50 alfalfa hay:concentrate diet

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N. Khateri

2017-03-01

Full Text Available Objective An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO, containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control, 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results Ruminal pH, total volatile fatty acids (VFA concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05 compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05 in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion The results of the present study suggested that supplementation of MEO may have limited effects on apparent

The bacterial population adherent to plant particles in the rumen of reindeer fed lichen, timothy hay or silage

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Monica Alterskjær Olsen

1998-02-01

Full Text Available Male reindeer (Rangifer tarandus tarandus calves taken from a natural winter pasture were given ad lib. access to lichen (n = 3, timothy silage (n = 3 and hay (n = 3 for 7 weeks. Median numbers of viable anaerobic bacteria adherent to the plant particles (cells/g wet weight of rumen solids, growing on a habitat simulating medium (M8V, were significantly higher (P = 0.05 in the rumen of reindeer fed lichen (26.5 x 109- 53.0 x 109 and hay (4.0 x 109- 40.5 x 109, compared to reindeer fed silage (1.15 x 109 - 3.25 x 109. Anaerobic bacterial strains (n = 551 from the plant particles obtained from the rumen of the nine reindeer examined, were isolated using an acid swollen cellulose medium (M8SC and tested for their ability to hydrolyse carboxymethyl cellulose (CMC. The proportion of CMC hydrolysing adherent bacteria isolated from M8SC was significantly higher in reindeer fed hay (21.5% compared ro animals fed lichen (5.3% and silage (2.7% (P = 0.05. The CMC hydrolysing bacterial srrains (n=42 isolated from reindeer fed hay where characterised as non-cellulolytic Butyrivibrio fibrisolvens (9.5%, cellulolytic B. fibrisolvens (50.0%, Clostridium sp. (2.4% and unknowns (38.1%, while CMC hydrolysing strains (n=11 isolated from animals fed lichen and strains (n=4 isolated from animals fed silage where all characterised as B. fibrisolvens. None of the bacterial strains isolated from the rumen solids of reindeer fed lichen or silage were found to be cellulolytic. This study suggests that both lichen and timothy silage have a negative influence, compared to hay, on the numbers of cellulolytic bacteria adherent to the plant particles in the rumen of reindeer.

Changes in the rumen bacterial microbiome of cattle exposed to ponderosa pine needles.

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Welch, K D; Stonecipher, C A; Gardner, D R; Cook, D; Pfister, J A

2017-05-01

Consumption of ponderosa pine needles, as well as needles and bark from a number of other trees, can cause abortions in cattle. The abortifacient compounds in these trees are labdane resin acids, including isocupressic acid and agathic acid. Previous research has demonstrated that cattle conditioned to pine needles metabolize the labdane resin acids more quickly than naïve cattle. The results from that study indicated that changes had occurred in the rumen of conditioned cattle. Therefore, in this study, the changes that occurred in the rumen bacterial microflora of cattle during exposure to ponderosa pine needles were evaluated. Cattle were dosed with ground pine needles twice daily for 7 d. Rumen samples were collected on d 0, 3, 7, and 14 (7 d after treatment stopped) and ruminal bacterial microbiome analyses were performed. There were 372 different genera of bacteria identified in the rumen samples. Principal coordinate analysis indicated that there was a significant difference in the rumen bacterial composition between the time points. There were 18 genera that increased in abundance from d 0 to d 7. Twenty three genera decreased in abundance from d 0 to d 7. The results from this study demonstrated that exposure of cattle to pine needles caused a clear shift in the rumen microbiome composition. In general, this shift lasted less than 1 wk post exposure, which indicates that any prophylactic treatment to manipulate the ruminal metabolism of the abortifacient compounds in pine needles would need to be continuously administered to maintain the necessary microbial composition in the rumen.

microbiology of the rumen in relation to the nutrition and physiology

African Journals Online (AJOL)

Removal of undigested solid residues. Removal of gas. ... activities of the rumen bacteria in so far as they can vary ... a means whereby the amount of any given substrate in the diet and the ... from starch and sucrose, the fermentation of these sub- ..... Functional ruminal flora and ruminal levels of lactic and volatile fatty acids.

Isolation and characterization of feather degrading bacteria from ...

African Journals Online (AJOL)

This study is aimed at isolating and characterizing new culturable feather degrading bacteria from soils of the University of Mauritius Farm. Bacteria that were isolated were tested for their capability to grow on feather meal agar (FMA). Proteolytic bacteria were tested for feather degradation and were further identified ...

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

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Ding, Gengzhi; Chang, Ying; Zhao, Liping; Zhou, Zhenming; Ren, Liping; Meng, Qingxiang

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 × 10(9) 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 1(st) to the 4(th) 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 trend for these parameters. SC supplementation significantly (P 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 higher (P < 0.05) in the SC group. In a word, dietary CTFR had a significant effect on degradation characteristics of forage and rumen microbial population. S. cerevisiae had positive effects on DM and NDF degradation rate or effective degradability of forage; S. cerevisiae increased rumen total bacteria, fungi, protozoa, and lactate-utilizing bacteria but reduced

Ammonia production by human faecal bacteria, and the enumeration, isolation and characterization of bacteria capable of growth on peptides and amino acids

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Richardson Anthony J

2013-01-01

Full Text Available Abstract Background The products of protein breakdown in the human colon are considered to be detrimental to gut health. Amino acid catabolism leads to the formation of sulfides, phenolic compounds and amines, which are inflammatory and/or precursors to the formation of carcinogens, including N-nitroso compounds. The aim of this study was to investigate the kinetics of protein breakdown and the bacterial species involved. Results Casein, pancreatic casein hydrolysate (mainly short-chain peptides or amino acids were incubated in vitro with suspensions of faecal bacteria from 3 omnivorous and 3 vegetarian human donors. Results from the two donor groups were similar. Ammonia production was highest from peptides, followed by casein and amino acids, which were similar. The amino acids metabolized most extensively were Asp, Ser, Lys and Glu. Monensin inhibited the rate of ammonia production from amino acids by 60% (P = 0.001, indicating the involvement of Gram-positive bacteria. Enrichment cultures were carried out to investigate if, by analogy with the rumen, there was a significant population of asaccharolytic, obligately amino acid-fermenting bacteria (‘hyper-ammonia-producing’ bacteria; HAP in the colon. Numbers of bacteria capable of growth on peptides or amino acids alone averaged 3.5% of the total viable count, somewhat higher than the rumen. None of these were HAP, however. The species enriched included Clostridium spp., one of which was C. perfringens, Enterococcus, Shigella and Escherichia coli. Conclusions Protein fermentation by human faecal bacteria in the absence of sugars not only leads to the formation of hazardous metabolic products, but also to the possible proliferation of harmful bacteria. The kinetics of protein metabolism were similar to the rumen, but HAP bacteria were not found.

The relationship between odd- and branched-chain fatty acids and microbial nucleic acid bases in rumen.

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Liu, Keyuan; Hao, Xiaoyan; Li, Yang; Luo, Guobin; Zhang, Yonggen; Xin, Hangshu

2017-11-01

This study aims to identify the relationship between odd- and branched-chain fatty acids (OBCFAs) and microbial nucleic acid bases in the rumen, and to establish a model to accurately predict microbial protein flow by using OBCFA. To develop the regression equations, data on the rumen contents of individual cows were obtained from 2 feeding experiments. In the first experiment, 3 rumen-fistulated dry dairy cows arranged in a 3×3 Latin square were fed diets of differing forage to concentration ratios (F:C). The second experiment consisted of 9 lactating Holstein dairy cows of similar body weights at the same stage of pregnancy. For each lactation stage, 3 cows with similar milk production were selected. The rumen contents were sampled at 4 time points of every two hours after morning feeding 6 h, and then to analyse the concentrations of OBCFA and microbial nucleic acid bases in the rumen samples. The ruminal bacteria nucleic acid bases were significantly influenced by feeding diets of differing forge to concentration ratios and lactation stages of dairy cows (pacids and C15:0 isomers, strongly correlated with the microbial nucleic acid bases in the rumen (pacid bases established by ruminal OBCFAs contents showed a good predictive capacity, as indicated by reasonably low standard errors and high R-squared values. This finding suggests that the rumen OBCFA composition could be used as an internal marker of rumen microbial matter.

Proteolytic and ACE-inhibitory activities of probiotic yogurt containing non-viable bacteria as affected by different levels of fat, inulin and starter culture.

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Shakerian, Mansour; Razavi, Seyed Hadi; Ziai, Seyed Ali; Khodaiyan, Faramarz; Yarmand, Mohammad Saeid; Moayedi, Ali

2015-04-01

In this study, the effects of fat (0.5 %, 3.2 % and 5.0 %), inulin (0.0 and 1.0 %) and starter culture (0.0 %, 0.5 %, 1.0 % and 1.5 %) on the angiotensin converting enzyme (ACE)-inhibitory activity of probiotic yogurt containing non-viable bacteria were assessed. Proteolytic activities of bacteria were also investigated. Yogurts were prepared either using a sole yogurt commercial culture including Streptococcus thermophilus and Lactobacillus delbrueckii subs. bulgaricus or bifidobacterium animalis BB-12 and Lactobacillus acidophilus La5 in addition to yogurt culture. Relative degrees of proteolysis were found to be considerably higher in yogurt samples than UHT milk as the control. Both regular and probiotic yogurts showed considerable ACE-inhibitory activities. Results showed that degree of proteolysis was not influenced by different fat contents, while was increased by high concentration of starter culture (1.5 % w/w) and reduced by inulin (1 % w/w). ACE-inhibitory activities of yogurt were also negatively affected by the presence of inulin and high levels of fat (5 % w/w). Moreover, yogurt containing probiotic bacteria showed higher inhibitory against ACE in comparison to the yogurt prepared with non-probiotic strains.

Minerals and rumen function

International Nuclear Information System (INIS)

Smith, R.H.

1984-01-01

The mechanisms are discussed of some clinical disorders, characteristic only of ruminants and related to the effects of abnormal mineral intake on rumen function. With particular attention to tropical conditions, consideration is given to: (a) the possible effects of phosphorus deficiency on rumen microbial activity; (b) the depression of rumen microbial synthesis in sulphur deficiency; (c) the inhibition of magnesium absorption from the forestomachs; and (d) the involvement of the rumen microorganisms in leading to copper and vitamin B 12 deficiencies as a result of low intakes of cobalt. (author)

Studies on fibrolytic bacterium Butyrivibrio fibrisolvens isolated from sheep rumen

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Sawanon, S.

2007-03-01

Full Text Available Fibrolytic Butyrivibrio fibrisolvens was an attractive target for genetic engineering in rumen bacteria. The experiment was initiated in making culture collection of this species, some of which may be useful ascandidate strain in the future. Hay suspended in sheep rumen was used as the source of isolates. The source was enriched with filter paper degradation, diluted with an anaerobic solution and used for pure culturing bya roll tube technique. After colony forming, Gram-negative curved rods bacteria were selected and screened for further identification with volatile fatty acid (VFA profiling and 16S rDNA sequencing. Fibrolyticstrains were selected to find fibrolytic enzymes and attachment to and digestion of various fibers. Fortyseven strains of Gram-negative curved rods were isolated. After determining cellulase, xylanase activities and VFA profile, 2 strains were chosen and employed for 16S rDNA sequencing. Both strains producingbutyrate were B. fibrisolvens. Of these 2 strains, most fibrolytic S-28 was selected. The strain S-28 could degrade natural fibers but not cellulose and showed strong attachment to them. A strong xylanase activitywas detected and presence of cellulase, β-glucosidase, β-xylosidase, α-L-arabinofuranosidase and β- cellobiosidase were also demonstrated.

Role of Age-Related Shifts in Rumen Bacteria and Methanogens in Methane Production in Cattle

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Chong Liu

2017-08-01

Full Text Available Rumen microbiota are essential for maintaining digestive and metabolic functions, producing methane as a byproduct. Dairy heifers produce large amounts of methane based on fermentation of digested organic matter, with adverse consequences for feed efficiency and the environment. It is therefore important to understand the influence of host age on the relationship between microbiota and methane production. This study explored the age effect on the relationship between microbial communities and enteric methane production in dairy cows and heifers using high-throughput sequencing. Methane production and volatile fatty acid concentrations were age-related. Heifers (9–10 months had lower methane production but higher methane production per dry matter intake (DMI. The acetate:propionate ratio decreased significantly with increasing age. Age-related microbiota changes in the rumen were reflected by a significant shift in bacterial taxa, but relatively stable archaeal taxa. Prevotella, Ruminococcus, Flavonifractor, Succinivibrio, and Methanobrevibacter were affected by age. This study revealed different associations between predominant bacterial phylotypes and Methanobrevibacter with increasing age. Prevotella was strongly correlated with Methanobrevibacter in heifers; howerver, in older cows (96–120 months this association was replaced by a correlation between Succinivibrio and Methanobrevibacter. This shift may account for the age-related difference in rumen fermentation and methane production per DMI.

Uji Biologis Konsumsi Pakan, Populasi Bakteri Rumen dan pH Pellet Complete Calf Starter pada Pedet Friesian Holstein Pra Sapih

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Nadia Maharani

2015-04-01

Full Text Available (Biological test feed intake, population rumen bacteria and ph pellet complete calf starter friesian holstein on pre weaning calf  ABSTRACT. The study aims to assess the quality of the formula Complete Calf Starter (CCS with 5% molasses instead of milk to the development of rumen microbial calf Holstein Friesian (HF pre-weaning. The material used is 20 head calf FH pre weaning age of 2 weeks. Feed intake data taken from 20 respondents consisted of 16 females and 4 calf tail male calf. Data rumen bacterial populations and pH were taken from 5 head of cattle slaughtered at the age of 2 weeks (one tail, 4 weeks (2-tailed and 6 weeks (2-tailed. The study was designed as a descriptive non-parametric. Parameters measured were rumen bacterial populations, pH and feed intake. The results showed the average consumption of dry matter (DM and CCS calf milk FH ages 2, 4, 6 weeks respectively at 506, 517, 528 grams. Rumen bacterial populations in calves aged 2, 4, and 6 weeks of 80x106, 45x106 and 19x106 kol/m. ruminal pH in calves aged 2, 4 and 6 weeks was 5.71, 5.36 and 5.55. Biological test showed that the use of complete calf starter (CCS in the form of pellets FH calves at the age of 2 weeks to stimulate the development of the rumen (feed intake, the population of bacteria and pH but not in calves aged 4 and 6 weeks. This is because the number of bacterial populations declined due to impaired absorption is impaired absorption of VFA marked low rumen pH.

Associative patterns among anaerobic fungi, methanogenic archaea, and bacterial communities in response to changes in diet and age in the rumen of dairy cows.

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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 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 age.

Insights into resistome and stress responses genes in Bubalus bubalis rumen through metagenomic analysis.

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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.

The relationship between odd- and branched-chain fatty acids and microbial nucleic acid bases in rumen

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Keyuan Liu

2017-11-01

Full Text Available Objective This study aims to identify the relationship between odd- and branched-chain fatty acids (OBCFAs and microbial nucleic acid bases in the rumen, and to establish a model to accurately predict microbial protein flow by using OBCFA. Methods To develop the regression equations, data on the rumen contents of individual cows were obtained from 2 feeding experiments. In the first experiment, 3 rumen-fistulated dry dairy cows arranged in a 3×3 Latin square were fed diets of differing forage to concentration ratios (F:C. The second experiment consisted of 9 lactating Holstein dairy cows of similar body weights at the same stage of pregnancy. For each lactation stage, 3 cows with similar milk production were selected. The rumen contents were sampled at 4 time points of every two hours after morning feeding 6 h, and then to analyse the concentrations of OBCFA and microbial nucleic acid bases in the rumen samples. Results The ruminal bacteria nucleic acid bases were significantly influenced by feeding diets of differing forge to concentration ratios and lactation stages of dairy cows (p<0.05. The concentrations of OBCFAs, especially odd-chain fatty acids and C15:0 isomers, strongly correlated with the microbial nucleic acid bases in the rumen (p<0.05. The equations of ruminal microbial nucleic acid bases established by ruminal OBCFAs contents showed a good predictive capacity, as indicated by reasonably low standard errors and high R-squared values. Conclusion This finding suggests that the rumen OBCFA composition could be used as an internal marker of rumen microbial matter.

Isolation of high-quality total RNA from rumen anaerobic bacteria and fungi, and subsequent detection of glycoside hydrolases.

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Wang, Pan; Qi, Meng; Barboza, Perry; Leigh, Mary Beth; Ungerfeld, Emilio; Selinger, L Brent; McAllister, Tim A; Forster, Robert J

2011-07-01

The rumen is one of the most powerful fibrolytic fermentation systems known. Gene expression analyses, such as reverse transcription PCR (RT-PCR), microarrays, and metatranscriptomics, are techniques that could significantly expand our understanding of this ecosystem. The ability to isolate and stabilize representative RNA samples is critical to obtaining reliable results with these procedures. In this study, we successfully isolated high-quality total RNA from the solid phase of ruminal contents by using an improved RNA extraction method. This method is based on liquid nitrogen grinding of whole ruminal solids without microbial detachment and acid guanidinium - phenol - chloroform extraction combined with column purification. Yields of total RNA were as high as 150 µg per g of fresh ruminal content. The typical large subunit/small subunit rRNA ratio ranged from 1.8 to 2.0 with an RNA integrity number (Agilent Technologies) greater than 8.5. By eliminating the detachment step, the resulting RNA was more representative of the complete ecosystem. Our improved method removed a major barrier limiting analysis of rumen microbial function from a gene expression perspective. The polyA-tailed eukaryotic mRNAs obtained have successfully been applied to next-generation sequencing, and metatranscriptomic analysis of the solid fraction of rumen contents revealed abundant sequences related to rumen fungi.

Effect of bioaugmentation by cellulolytic bacteria enriched from sheep rumen on methane production from wheat straw.

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Ozbayram, E Gozde; Kleinsteuber, Sabine; Nikolausz, Marcell; Ince, Bahar; Ince, Orhan

2017-08-01

The aim of this study was to determine the potential of bioaugmentation with cellulolytic rumen microbiota to enhance the anaerobic digestion of lignocellulosic feedstock. An anaerobic cellulolytic culture was enriched from sheep rumen fluid using wheat straw as substrate under mesophilic conditions. To investigate the effects of bioaugmentation on methane production from straw, the enrichment culture was added to batch reactors in proportions of 2% (Set-1) and 4% (Set-2) of the microbial cell number of the standard inoculum slurry. The methane production in the bioaugmented reactors was higher than in the control reactors. After 30 days of batch incubation, the average methane yield was 154 mL N CH 4 g VS -1 in the control reactors. Addition of 2% enrichment culture did not enhance methane production, whereas in Set-2 the methane yield was increased by 27%. The bacterial communities were examined by 454 amplicon sequencing of 16S rRNA genes, while terminal restriction fragment length polymorphism (T-RFLP) fingerprinting of mcrA genes was applied to analyze the methanogenic communities. The results highlighted that relative abundances of Ruminococcaceae and Lachnospiraceae increased during the enrichment. However, Cloacamonaceae, which were abundant in the standard inoculum, dominated the bacterial communities of all batch reactors. T-RFLP profiles revealed that Methanobacteriales were predominant in the rumen fluid, whereas the enrichment culture was dominated by Methanosarcinales. In the batch rectors, the most abundant methanogens were affiliated to Methanobacteriales and Methanomicrobiales. Our results suggest that bioaugmentation with sheep rumen enrichment cultures can enhance the performance of digesters treating lignocellulosic feedstock. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of Nitrooxy Compounds with Different Molecular Structures on the Rumen Methanogenesis, Metabolic Profile, and Methanogenic Community.

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Jin, Wei; Meng, Zhenxiang; Wang, Jing; Cheng, Yanfen; Zhu, Weiyun

2017-08-01

Rumen in vitro fermentation was used to evaluate the capacity of nitrooxy compounds to mitigate rumen methane production. The following three nitrooxy compounds, each with different molecular structures, were evaluated: 2,2-dimethyl-3-(nitrooxy) propanoic (DNP), N-[2-(Nitrooxy)ethyl]-3-pyridinecarboxamide (NPD), and nitroglycerin (NG). All three compounds substantially decreased the total gas production, methane production, and the acetate:propionate ratio, while increasing hydrogen production. The growth of methanogens was specifically inhibited by all three compounds, without affecting the abundance of bacteria, anaerobic fungi, or protozoa. However, inhibition of methanogenesis required a much higher dose of DNP when compared to NPD or NG. Further investigations were conducted on NG to determine its effects on the methanogenic community. NG reduced the relative abundance of Methanomassiliicoccales, while increasing the relative abundance of Methanobrevibacter and Methanosphaera. Overall, the results suggested that all three of these nitrooxy compounds could specifically inhibit rumen methanogenesis, but NPD and NG were much more efficient than DNP at rumen methane mitigation.

Technical note: A simple rumen collection device for calves: An adaptation of a manual rumen drenching system.

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Klopp, R N; Oconitrillo, M J; Sackett, A; Hill, T M; Schlotterbeck, R L; Lascano, G J

2018-07-01

A limited amount of research is available related to the rumen microbiota of calves, yet there has been a recent spike of interest in determining the diversity and development of calf rumen microbial populations. To study the microbial populations of a calf's rumen, a sample of the rumen fluid is needed. One way to take a rumen fluid sample from a calf is by fistulating the animal. This method requires surgery and can be very stressful on a young animal that is trying to adapt to a new environment and has a depressed immune system. Another method that can be used instead of fistulation surgery is a rumen pump. This method requires a tube to be inserted into the rumen through the calf's esophagus. Once inside the rumen, fluid can be pumped out and collected in a few minutes. This method is quick, inexpensive, and does not cause significant stress on the animal. This technical note presents the materials and methodology used to convert a drenching system into a rumen pump and its respective utilization in 2 experiments using dairy bull calves. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Deteksi Keragaman Spesies Bakteri Metanogen Rumen Sapi Menggunakan Kloning Gen 16s Rrna dan Sekuensing

OpenAIRE

Noor, Shoffiana; Pramono, Hendro; Aziz, Saefuddin

2014-01-01

Ruminants produce methane gas which contributes to enhanced greenhouse effect in the atmosphere. Cattle issued the highest methane during the fermentation of feed in the rumen. Methane gas produced by methanogen bacteria in carbohydrates anaerobic fermentation. Methanogen bacteria are difficult to obtain diversity information because difficult cultured. One technique can be used is molecular rRNA 16S gene cloning and sequencing. This study was aims to determine the species diversity of methan...

Degradation of spent craft brewer’s yeast by caprine rumen hyper ammonia-producing bacteria

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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...

A study of nutritional status of Finnish reindeer (Rangifer Tarandus L. in differents months: I. Composition and volume of the rumen microbiota

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Liisa Syrjälä

1973-12-01

Full Text Available The rumen microbiota were studied in free-ranging semi-domestic reindeer in Finnish Lapland under the nutritional conditions obtaining at two different sampling times. Qualitative and quantitative investigations were made of the rumen ciliate fauna and quantitative investigations of the rumen bacterial flora. The volume coefficients for rumen ciliates obtained by Westerling (1970 and that for rumen bacteria obtained by Warner (1962 were used to obtain an indication of the volume of the rumen microbe mass in reindeer. The rumen samples were collected in connection with the round-up and slaughter of reindeer, being taken from 30 animals in December and 29 animals in March. The reindeer slaughtered in December had normal access to food, but those slaughtered in March had grazed on better pastures and received a supplementary feed of hay. The total number of ciliate cells was over six times as high in March as in December, the numbers being 1 182900 and 188 300 per ml rumen contents, respectively. The corresponding total numbers of bacterial cells were 9.65 x 109 in March and 6.65 x 109 in December. The reason for the statistically significantly (P < 0.01 higher numbers in March than in December is probably the better nutritional conditions of the herd slaughtered in March, not the time of the year. The ciliate fauna consisted of 19 different species, although not all the species were found in every sample. The percentage composition of the ciliate fauna did not vary considerably between the two sampling times. The volume of the total microbe mass constituted 8.2% of the rumen contents in March and 1.9 % in December, the average being 5.1 %. The proportion of the ciliate volume in the total microbe mass was clearly higher than that of the bacteria at both sampling times: 7.2 times as high in March and 1.7 times in December, the average being 4.7 times.

Role of anaerobic fungi in wheat straw degradation and effects of plant feed additives on rumen fermentation parameters in vitro.

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Dagar, S S; Singh, N; Goel, N; Kumar, S; Puniya, A K

2015-01-01

In the present study, rumen microbial groups, i.e. total rumen microbes (TRM), total anaerobic fungi (TAF), avicel enriched bacteria (AEB) and neutral detergent fibre enriched bacteria (NEB) were evaluated for wheat straw (WS) degradability and different fermentation parameters in vitro. Highest WS degradation was shown for TRM, followed by TAF, NEB and least by AEB. Similar patterns were observed with total gas production and short chain fatty acid profiles. Overall, TAF emerged as the most potent individual microbial group. In order to enhance the fibrolytic and rumen fermentation potential of TAF, we evaluated 18 plant feed additives in vitro. Among these, six plant additives namely Albizia lebbeck, Alstonia scholaris, Bacopa monnieri, Lawsonia inermis, Psidium guajava and Terminalia arjuna considerably improved WS degradation by TAF. Further evaluation showed A. lebbeck as best feed additive. The study revealed that TAF plays a significant role in WS degradation and their fibrolytic activities can be improved by inclusion of A. lebbeck in fermentation medium. Further studies are warranted to elucidate its active constituents, effect on fungal population and in vivo potential in animal system.

Changes in Microbiota in Rumen Digesta and Feces Due to a Grain-Based Subacute Ruminal Acidosis (SARA) Challenge

DEFF Research Database (Denmark)

Plaizier, Jan C.; Li, Shucong; Danscher, Anne Mette

2017-01-01

The effects of a grain-based subacute ruminal acidosis (SARA) challenge on bacteria in the rumen and feces of lactating dairy cows were determined. Six lactating, rumen-cannulated Danish Holstein cows were used in a cross-over study with two periods. Periods included two cows on a control diet...... and two cows on a SARA challenge. The control diet was a total mixed ration containing 45.5% dry matter (DM), 43.8% DM neutral detergent fiber, and 19.6% DM starch. The SARA challenge was conducted by gradually substituting the control diet with pellets containing 50% wheat and 50% barley over 3 days...... to reach a diet containing 55.6% DM, 31.3% DM neutral detergent fiber, and 31.8% DM starch, which was fed for four more days. Rumen fluid samples were collected at day 7 and 10 of experimental periods. Feces samples were collected on days 8 and 10 of these periods. Extracted DNA from the rumen and feces...

The Influence of Tallow on Rumen Metabolism, Microbial Biomass Synthesis and Fatty Acid Composition of Bacteria and Protozoa

DEFF Research Database (Denmark)

Weisbjerg, Martin Riis; Børsting, Christian Friis; Hvelplund, Torben

1992-01-01

Rumen metabolism, microbial biomass synthesis and microbial long chain fatty acid composition were studied in lactating cows fed at two levels of dry matter intake (L, 8.6 kg DM and H, 12.6 kg DM) with 0, 4 and 6% added tallow at the low feed level (L0, L4 and L6) and 0, 2, 4 and 6% at the high...... feed level (H0, H2, H4 and H6). Fibre digestibility was not significantly affected by tallow addition. Increasing tallow level in the diet decreased the total VFA concentration, the ratio of acetic acid to propionic acid and the ammonia concentration in the rumen. Crude fat and fatty acid content...... in bacterial and protozoal dry matter increased with increased tallow level, especially due to an increase in fatty acids originating from the feeds. Microbial synthesis in the rumen and flow of amino acids to the duodenum was highest for medium fat intake at the high feed level....

Short communication: Comparison of pH, volatile fatty acids, and microbiome of rumen samples from preweaned calves obtained via cannula or stomach tube.

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Terré, M; Castells, L; Fàbregas, F; Bach, A

2013-08-01

The objective of this study was to compare rumen samples from young dairy calves obtained via a stomach tube (ST) or a ruminal cannula (RC). Five male Holstein calves (46±4.0 kg of body weight and 11±4.9 d of age) were ruminally cannulated at 15 d of age. Calves received 4 L/d of a commercial milk replacer (25% crude protein and 19.2% fat) at 12.5% dry matter, and were provided concentrate and chopped oats hay ad libitum throughout the study (56 d). In total, 29 paired rumen samples were obtained weekly throughout the study in most of the calves by each extraction method. These samples were used to determine pH and volatile fatty acids (VFA) concentration, and to quantify Prevotella ruminicola and Streptococcus bovis by quantitative PCR. Furthermore, a denaturing gradient gel electrophoresis was performed on rumen samples harvested during wk 8 of the study to determine the degree of similarity between rumen bacteria communities. Rumen pH was 0.30 units greater in ST compared with RC samples. Furthermore, total VFA concentrations were greater in RC than in ST samples. However, when analyzing the proportion of each VFA by ANOVA, no differences were found between the sampling methods. The quantification of S. bovis and P. ruminicola was similar in both extraction methods, and values obtained using different methods were highly correlated (R(2)=0.89 and 0.98 for S. bovis and P. ruminicola, respectively). Fingerprinting analysis showed similar bacteria band profiles between samples obtained from the same calves using different extraction methods. In conclusion, when comparing rumen parameters obtained using different sampling techniques, it is recommended that VFA profiles be used rather than total VFA concentrations, as total VFA concentrations are more affected by the method of collection. Furthermore, although comparisons of pH across studies should be avoided when samples are not obtained using the same sampling method, the comparison of fingerprinting of a

Metagenomic insights into the rumen microbial fibrolytic enzymes in Indian crossbred cattle fed finger millet straw.

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Jose, V Lyju; Appoothy, Thulasi; More, Ravi P; Arun, A Sha

2017-12-01

The rumen is a unique natural habitat, exhibiting an unparalleled genetic resource of fibrolytic enzymes of microbial origin that degrade plant polysaccharides. The objectives of this study were to identify the principal plant cell wall-degrading enzymes and the taxonomic profile of rumen microbial communities that are associated with it. The cattle rumen microflora and the carbohydrate-active enzymes were functionally classified through a whole metagenomic sequencing approach. Analysis of the assembled sequences by the Carbohydrate-active enzyme analysis Toolkit identified the candidate genes encoding fibrolytic enzymes belonging to different classes of glycoside hydrolases(11,010 contigs), glycosyltransferases (6366 contigs), carbohydrate esterases (4945 contigs), carbohydrate-binding modules (1975 contigs), polysaccharide lyases (480 contigs), and auxiliary activities (115 contigs). Phylogenetic analysis of CAZyme encoding contigs revealed that a significant proportion of CAZymes were contributed by bacteria belonging to genera Prevotella, Bacteroides, Fibrobacter, Clostridium, and Ruminococcus. The results indicated that the cattle rumen microbiome and the CAZymes are highly complex, structurally similar but compositionally distinct from other ruminants. The unique characteristics of rumen microbiota and the enzymes produced by resident microbes provide opportunities to improve the feed conversion efficiency in ruminants and serve as a reservoir of industrially important enzymes for cellulosic biofuel production.

Exploitation of dietary tannins to improve rumen metabolism and ruminant nutrition.

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Patra, Amlan K; Saxena, Jyotisna

2011-01-15

Tannins (hydrolysable and condensed tannin) are polyphenolic polymers of relatively high molecular weight with the capacity to form complexes mainly with proteins due to the presence of a large number of phenolic hydroxyl groups. They are widely distributed in nutritionally important forage trees, shrubs and legumes, cereals and grains, which are considered as anti-nutritional compounds due to their adverse effects on intake and animal performance. However, tannins have been recognised to modulate rumen fermentation favourably such as reducing protein degradation in the rumen, prevention of bloat, inhibition of methanogenesis and increasing conjugated linoleic acid concentrations in ruminant-derived foods. The inclusion of tannins in diets has been shown to improve body weight and wool growth, milk yields and reproductive performance. However, the beneficial effects on rumen modulation and animal performance have not been consistently observed. This review discusses the effects of tannins on nitrogen metabolism in the rumen and intestine, and microbial populations (bacteria, protozoa, fungi and archaea), metabolism of tannins, microbial tolerance mechanisms to tannins, inhibition of methanogenesis, ruminal biohydrogenation processes and performance of animals. The discrepancies of responses of tannins among different studies are attributed to the different chemical structures (degree of polymerisation, procyanidins to propdelphinidins, stereochemistry and C-C bonding) and concentrations of tannins, and type of diets. An establishment of structure-activity relationship would be required to explain differences among studies and obtain consistent beneficial tannin effects. Copyright © 2010 Society of Chemical Industry.

Rumen and Cecum Microbiomes in Reindeer (Rangifer tarandus tarandus) Are Changed in Response to a Lichen Diet and May Affect Enteric Methane Emissions.

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Salgado-Flores, Alejandro; Hagen, Live H; Ishaq, Suzanne L; Zamanzadeh, Mirzaman; Wright, André-Denis G; Pope, Phillip B; Sundset, Monica A

2016-01-01

Reindeer (Rangifer tarandus tarandus) are large Holarctic herbivores whose heterogeneous diet has led to the development of a unique gastrointestinal microbiota, essential for the digestion of arctic flora, which may include a large proportion of lichens during winter. Lichens are rich in plant secondary metabolites, which may affect members of the gut microbial consortium, such as the methane-producing methanogenic archaea. Little is known about the effect of lichen consumption on the rumen and cecum microbiotas and how this may affect methanogenesis in reindeer. Here, we examined the effects of dietary lichens on the reindeer gut microbiota, especially methanogens. Samples from the rumen and cecum were collected from two groups of reindeer, fed either lichens (Ld: n = 4), or a standard pelleted feed (Pd: n = 3). Microbial densities (methanogens, bacteria and protozoa) were quantified using quantitative real-time PCR and methanogen and bacterial diversities were determined by 454 pyrosequencing of the 16S rRNA genes. In general, the density of methanogens were not significantly affected (p>0.05) by the intake of lichens. Methanobrevibacter constituted the main archaeal genus (>95% of reads), with Mbr. thaueri CW as the dominant species in both groups of reindeer. Bacteria belonging to the uncharacterized Ruminococcaceae and the genus Prevotella were the dominant phylotypes in the rumen and cecum, in both diets (ranging between 16-38% total sequences). Bacteria belonging to the genus Ruminococcus (3.5% to 0.6%; p = 0.001) and uncharacterized phylotypes within the order Bacteroidales (8.4% to 1.3%; p = 0.027), were significantly decreased in the rumen of lichen-fed reindeer, but not in the cecum (p = 0.2 and p = 0.087, respectively). UniFrac-based analyses showed archaeal and bacterial libraries were significantly different between diets, in both the cecum and the rumen (vegan::Adonis: pseudo-Flichen-fed reindeer.

Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen.

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Denman, Stuart E; McSweeney, Christopher S

2006-12-01

Traditional methods for enumerating and identifying microbial populations within the rumen can be time consuming and cumbersome. Methods that involve culturing and microscopy can also be inconclusive, particularly when studying anaerobic rumen fungi. A real-time PCR SYBR Green assay, using PCR primers to target total rumen fungi and the cellulolytic bacteria Ruminococcus flavefaciens and Fibrobacter succinogenes, is described, including design and validation. The DNA and crude protein contents with respect to the fungal biomass of both polycentric and monocentric fungal isolates were investigated across the fungal growth stages to aid in standard curve generation. The primer sets used were found to be target specific with no detectable cross-reactivity. Subsequently, the real-time PCR assay was employed in a study to detect these populations within cattle rumen. The anaerobic fungal target was observed to increase 3.6-fold from 0 to 12 h after feeding. The results also indicated a 5.4-fold increase in F. succinogenes target between 0 and 12 h after feeding, whereas R. flavefaciens was observed to maintain more or less consistent levels. This is the first report of a real-time PCR assay to estimate the rumen anaerobic fungal population.

In Vitro Rumen Fermentation and Anti Mastitis Bacterial Activity of Diet Containing Betel Leaf Meal (Piper betle L.

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A. A. Yamin

2013-08-01

Full Text Available The aims of this experiment was to study the inhibition effect of betel leaf meal (BLM addition into concentrate diet on mastitis causing bacteria and on rumen fermentation condition. The study consisted of five dietary treatments of BLM level in concentrate feed, i.e., 0%, 2%, 4%, 6%, and 8% and four replicates of each treatment. The treatment diets together with napier grass in ratio of 40 : 60 were fermented using rumen liquor. All treatments were examined their antibacterial activity before and after fermentation. After four hours fermentation, supernatant of each samples were analyzed for VFA, NH3, number of bacteria and protozoa. Dry matter (DM and organic matter (OM digestibility were analyzed after 48 h fermentation. The results showed that before fermentation, 8% BLM addition caused the bigest (P<0.05 inhibition diameter of Staphylococcus spp. growth compared to other lower levels. However after fermentation there were no significant differences among the addition levels of BLM. Two per cent of BLM addition produced higher VFA (P<0.05 than the other addition levels. Ammonia concentration, dry matter (DM and organic matter (OM digestibility were not different among the treatments. Addition of BLM significantly (P<0.01 decreased protozoa number, but did not affect bacterial count. It is concluded that the addition of 2% BLM in concentrate feed can be used effectively to inhibit the growth of mastitis causing bacteria (Staphylococcus spp. and does not disturb rumen fermentation condition.

Effects of Saccharomyces cerevisiae fermentation products on performance and rumen fermentation and microbiota in dairy cows fed a diet containing low quality forage

Institute of Scientific and Technical Information of China (English)

Wen Zhu; Zihai Wei; Ningning Xu; Fan Yang; Ilkyu Yoon; Yihua Chung; Jianxin Liu; Jiakun Wang

2017-01-01

Background:A possible option to meet the increased demand of forage for dairy industry is to use the agricultural byproducts,such as corn stover.However,nutritional value of crop residues is low and we have been seeking technologies to improve the value.A feeding trial was performed to evaluate the effects of four levels of Soccharomyces cerevisiae fermentation product (SCFP;Original XP;Diamond V) on lactation performance and rumen fermentation in mid-lactation Holstein dairy cows fed a diet containing low-quality forage.Eighty dairy cows were randomly assigned into one of four treatments:basal diet supplemented with 0,60,120,or 180 g/d of SCFP per head mixed with 180,120,60,or 0 g of corn meal,respectively.The experiment lasted for 10 wks,with the first 2 weeks for adaptation.Results:Dry matter intake was found to be similar (P ＞ 0.05) among the treatments.There was an increasing trend in milk production (linear,P ≤ 0.10) with the increasing level of SCFP supplementation,with no effects on contents of milk components (P ＞ 0.05).Supplementation of SCFP linearly increased (P ＜ 0.05) the N conversion,without affecting rumen pH and ammonia-N (P ＞ 0.05).Increasing level of SCFP linearly increased (P ＜ 0.05) concentrations of ruminal total volatile fatty acids,acetate,propionate,and butyrate,with no difference in molar proportion of individual acids (P ＞ 0.05).The population of fungi and certain cellulolytic bacteria (Ruminococcus albus,R.flavefaciens and Fibrobacter succinogenes)increased linearly (P ＜ 0.05) but those of lactate-utilizing (5elenomonas ruminontium and Megasphaera elsdenii) and lactate-producing bacteria (Streptococcus bovis) decreased linearly (P ≤ 0.01) with increasing level of SCFP.The urinary purine derivatives increased linearly (P ＜ 0.05) in response to SCFP supplementation,indicating that SCFP supplementation may benefit for microbial protein synthesis in the rumen.Conclusions:The SCFP supplementation was effective in

Intravenous lipid infusion affects dry matter intake, methane yield, and rumen bacteria structure in late-lactating Holstein cows.

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Lamp, Ole; Reyer, Henry; Otten, Winfried; Nürnberg, Gerd; Derno, Michael; Wimmers, Klaus; Metges, Cornelia C; Kuhla, Björn

2018-03-28

Increasing the dietary fat content of ruminant diets decreases methane (CH 4 ) production. This effect is caused by the toxic properties of fatty acids on rumen microbial populations, coating of feed particles diminishing the accessibility for microbes, and a reduction in dry matter intake (DMI). The latter effect is caused by postabsorptive long-chain fatty acids eliciting anorexic signaling; however, whether circulating long-chain fatty acids affect rumen CH 4 production alike is unknown. To approach this question, 5 rumen-cannulated Holstein cows in late lactation received 2 jugular catheters and were kept in respiration chambers to measure CH 4 production and DMI for 48 h. In a crossover design, cows were intravenously infused with a 20% lipid emulsion (LIPO) or 0.9% NaCl (CON). The LIPO cows received 2.1 kg of triglycerides/d [0.152 ± 0.007 g of triglycerides/(kg of BW × h) -1 ] consisting of 12.1% palmitic acid, 4.2% stearic acid, 31.1% oleic acid, and 52.7% linoleic acid. Blood and rumen fluid samples were taken hourly during the day. Results showed that LIPO compared with CON infusion increased plasma triglyceride as well as free fatty acid and serotonin concentrations but reduced the proportion of de novo synthesized milk fatty acids (sum of C6 to C16). Daily CH 4 production and DMI were lower, whereas daily CH 4 yield (CH 4 /DMI) was greater in LIPO than CON cows, although CH 4 yield decreased from d 1 to d 2 by 2 to 14% in LIPO-infused cows only. This effect was associated with a higher (acetate + butyrate)/propionate ratio, tending lower propionate concentrations between 24 and 34 h of infusion, reduced relative abundances of genera belonging to Succinivibrio, Ruminococcaceae, and Ruminiclostridium, and greater relative Bacteroidetes genus abundances in the rumen. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

In-depth diversity analysis of the bacterial community resident in the camel rumen.

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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. Copyright © 2014. Published by Elsevier GmbH.

Molecular diversity of the rumen microbiome of Norwegian reindeer on natural summer pasture.

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Sundset, Monica A; Edwards, Joan E; Cheng, Yan Fen; Senosiain, Roberto S; Fraile, Maria N; Northwood, Korinne S; Praesteng, Kirsti E; Glad, Trine; Mathiesen, Svein D; Wright, André-Denis G

2009-02-01

The molecular diversity of the rumen microbiome was investigated in five semi-domesticated adult female Norwegian reindeer (Rangifer tarandus tarandus) grazing on natural summer pastures on the coast of northern Norway (71.00 degrees N, 25.30 degrees E). Mean population densities (numbers per gram wet weight) of methanogenic archaea, rumen bacteria and ciliate protozoa, estimated using quantitative real-time polymerase chain reaction (PCR), were 3.17x10(9), 5.17x10(11) and 4.02x10(7), respectively. Molecular diversity of rumen methanogens was revealed using a 16S rRNA gene library (54 clones) constructed using pooled PCR products from the whole rumen contents of the five individual reindeer. Based upon a similarity criterion of rumen exhibited a high degree of sequence similarity to methanogens affiliated with the families Methanobacteriaceae (14 OTUs) and Methanosarcinaceae (one OTU). Four of the OTUs detected belonged to a group of uncultivated archaea previously found in domestic ruminants and thought to be dominant in the rumen together with Methanobrevibacter spp. Denaturing gradient gel electrophoresis profiling of the rumen bacterial 16S rRNA gene and the protozoal 18S rRNA gene indicated a high degree of animal variation, although some bands were common to all individuals. Automated ribosomal intergenic spacer analysis (ARISA) profiling of the ruminal Neocallimastigales population indicated that the reindeer are likely to contain more than one type of anaerobic fungus. The ARISA profile from one animal was distinct from the other four. This is the first molecular investigation of the ruminal methanogenic archaea in reindeer, revealing higher numbers than expected based on methane emission data available. Also, many of the reindeer archaeal 16S rRNA gene sequences were similar to those reported in domesticated ruminants in Australia, Canada, China, New Zealand and Venezuela, supporting previous findings that there seems to be no host type or geographical

Isolation of Pseudobutyrivibrio ruminis and Pseudobutyrivibrio xylanivorans from rumen of Creole goats fed native forage diet.

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Grilli, D J; Cerón, M E; Paez, S; Egea, V; Schnittger, L; Cravero, S; Escudero, M Sosa; Allegretti, L; Arenas, G N

2013-09-01

We isolated and identified functional groups of bacteria in the rumen of Creole goats involved in ruminal fermentation of native forage shrubs. The functional bacterial groups were evaluated by comparing the total viable, total anaerobic, cellulolytic, hemicellulolytic, and amylolytic bacterial counts in the samples taken from fistulated goats fed native forage diet (Atriplex lampa and Prosopis flexuosa). Alfalfa hay and corn were used as control diet. The roll tubes method increased the possibility of isolating and 16S rDNA gene sequencing allowed definitive identification of bacterial species involved in the ruminal fermentation. The starch and fiber contents of the diets influenced the number of total anaerobic bacteria and fibrolytic and amylolytic functional groups. Pseudobutyrivibrio ruminis and Pseudobutyrivibrio xylanivorans were the main species isolated and identified. The identification of bacterial strains involved in the rumen fermentation helps to explain the ability of these animals to digest fiber plant cell wall contained in native forage species.

Symbiosis and Rumen Protozoa

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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)

Hops (Humulus lupulus L. Bitter Acids: Modulation of Rumen Fermentation and Potential As an Alternative Growth Promoter

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Michael D. Flythe

2017-08-01

Full Text Available Antibiotics can improve ruminant growth and efficiency by altering rumen fermentation via selective inhibition of microorganisms. However, antibiotic use is increasingly restricted due to concerns about the spread of antibiotic-resistance. Plant-based antimicrobials are alternatives to antibiotics in animal production. The hops plant (Humulus lupulus L. produces a range of bioactive secondary metabolites, including antimicrobial prenylated phloroglucinols, which are commonly called alpha- and beta-acids. These latter compounds can be considered phyto-ionophores, phytochemicals with a similar antimicrobial mechanism of action to ionophore antibiotics (e.g., monensin, lasalocid. Like ionophores, the hop beta-acids inhibit rumen bacteria possessing a classical Gram-positive cell envelope. This selective inhibition causes several effects on rumen fermentation that are beneficial to finishing cattle, such as decreased proteolysis, ammonia production, acetate: propionate ratio, and methane production. This article reviews the effects of hops and hop secondary metabolites on rumen fermentation, including the physiological mechanisms on specific rumen microorganisms, and consequences for the ruminant host and ruminant production. Further, we propose that hop beta-acids are useful model natural products for ruminants because of (1 the ionophore-like mechanism of action and spectrum of activity and (2 the literature available on the plant due to its use in brewing.

Transformation and Stability of Cloned Polysaccharidase Genes in Gram-Positive Bacteria

OpenAIRE

EKİNCİ, Mehmet Sait

2014-01-01

Polysaccharidase genes from rumen bacteria were transferred to and expressed in ruminal and non-ruminal Gram-positive bacteria. The transformation efficiency and genetic stability of polysaccharidase genes in bacteria from different habitats were investigated. PCR amplification of cloned polysaccharidase genes from Escherichia coli, Lactococcus lactis, Enterococcus faecalis, Streptococcus sanguis and S. bovis strain 26 showed that rearrangement of plasmid and the gene fragment did not occur. ...

A scanning electron microscopy study of the invasion of leaflets of a bloat-safe and a bloat-causing legume by rumen microorganisms.

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Fay, J P; Cheng, K J; Hanna, M R; Howarth, R E; Costerton, J W

1981-04-01

A newly developed technique using ruthenium red to detect foci of bacterial digestion in mounts of whole leaflets that had been incubated with rumen bacteria was used to compare the digestion of alfalfa, a bloat-causing legume, and sainfoin, a bloat-safe legume. When whole leaflets were suspended in an artificial rumen medium and inoculated with rumen bacteria, massive bacterial adhesion and proliferation were noted at the stomata of alfalfa leaflets after 6 h of incubation, whereas only a few isolated bacteria adhered near the stomata of sainfoin leaflets After 22 h of incubation, the epidermal layers of alfalfa leaflets had peeled away in many areas, revealing an extensive bacterial invasion of the underlying mesophyll tissue in which large bacterial microcolonies had formed in intercellular spaces, and in intracellular spaces in several areas where plant cell walls had broken down. After 22 h of incubation, the surface of sainfoin leaflets resembled that of alfalfa leaflets at 6 h, with bacterial microcolonies adhering to the area surrounding the stomata, but without sloughing of the epidermis. Uninoculated control leaflets of both species showed no surface alteration but part of their normal bacterial flora had proliferated to form microcolonies on the surface after 22 h incubation. Dry matter loss due to leaching or bacterial digestion when whole leaflets of legumes were suspended in an artificial rumen medium, alone or with rumen bacteria, was significantly higher in the bloat-causing group. Values of leaching and of bacterial digestion were positively correlated. We conclude that reported differences in plant anatomy, and in cell wall chemistry, produce distinct rates or organic nutrient release from legume leaflets, and that these same differences produce an equally distinct susceptibility of leaflets to bacterial invasion, plant cell rupture, and the consequent release of intracellular plant components. The rate of release of organic nutrients from legume

Evaluation of the efficacy of rumen cannulation technique on some rumen metabolic parameters in buffaloes

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Gh. Manafiazar

2010-02-01

Full Text Available This experimental study was carried out to determine the efficacy of rumen cannulation technique on some rumen metabolic parameters in buffaloes. Four healthy male River (Azari buffaloes with no previous history of gastrointestinal dysfunction were chosen. There are several surgical techniques that can be used for rumen cannulation in farm animals, including buffaloes. This procedure was performed in a modified two-stage technique. In the first stage, the dorso – lateral portion of the dorsal sac of the rumen wall was sutured to the skin incision in the left para lumbar fossa region. In the second – stage, after six days left, the exposed rumen wall area was incised and the cannula was inserted and fixed manually in the rumenal opening incision site. In order to evaluate the efficacy of rumen cannulation technique on some rumen metabolic parameters, this study was achieved with different levels of NDF, and chewing behavior and their relationship with ruminal acidity, was measured in a change over design. Two diets with 2 levels of NDF were used as treatments. First and second diets had 52 and 47 % of NDF, respectively. Animals fed ad libitum at 09:00 and 21:00. There were no significant differences between chemical composition, particles distribution, geometric mean, its standard deviation and physically effective factor (pef of diets, dry matter intake (kg/d and nutrients intake (NDF, ADF, NFC and crude protein and their digestibility. Increasing NFC reduced ruminal pH at 0.5, 1.0, 4.5, 6.0 9.0 and 10.0 h post feeding. In addition, there were not significant differences on eating time, rumination time and total chewing activity between diets. All data obtained in this study were in normal range may indicating the efficacy of this cannulation method. More investigation should be done to determine the efficacy and comparison of the other surgical rumen cannulation techniques on Azari buffaloes of Iran.

Snapshot of the eukaryotic gene expression in muskoxen rumen--a metatranscriptomic approach.

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Meng Qi

Full Text Available BACKGROUND: Herbivores rely on digestive tract lignocellulolytic microorganisms, including bacteria, fungi and protozoa, to derive energy and carbon from plant cell wall polysaccharides. Culture independent metagenomic studies have been used to reveal the genetic content of the bacterial species within gut microbiomes. However, the nature of the genes encoded by eukaryotic protozoa and fungi within these environments has not been explored using metagenomic or metatranscriptomic approaches. METHODOLOGY/PRINCIPAL FINDINGS: In this study, a metatranscriptomic approach was used to investigate the functional diversity of the eukaryotic microorganisms within the rumen of muskoxen (Ovibos moschatus, with a focus on plant cell wall degrading enzymes. Polyadenylated RNA (mRNA was sequenced on the Illumina Genome Analyzer II system and 2.8 gigabases of sequences were obtained and 59129 contigs assembled. Plant cell wall degrading enzyme modules including glycoside hydrolases, carbohydrate esterases and polysaccharide lyases were identified from over 2500 contigs. These included a number of glycoside hydrolase family 6 (GH6, GH48 and swollenin modules, which have rarely been described in previous gut metagenomic studies. CONCLUSIONS/SIGNIFICANCE: The muskoxen rumen metatranscriptome demonstrates a much higher percentage of cellulase enzyme discovery and an 8.7x higher rate of total carbohydrate active enzyme discovery per gigabase of sequence than previous rumen metagenomes. This study provides a snapshot of eukaryotic gene expression in the muskoxen rumen, and identifies a number of candidate genes coding for potentially valuable lignocellulolytic enzymes.

Organic acid production from potato starch waste fermentation by rumen microbial communities from Dutch and Thai dairy cows.

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Palakawong Na Ayudthaya, Susakul; van de Weijer, Antonius H P; van Gelder, Antonie H; Stams, Alfons J M; de Vos, Willem M; Plugge, Caroline M

2018-01-01

Exploring different microbial sources for biotechnological production of organic acids is important. Dutch and Thai cow rumen samples were used as inocula to produce organic acid from starch waste in anaerobic reactors. Organic acid production profiles were determined and microbial communities were compared using 16S ribosomal ribonucleic acid gene amplicon pyrosequencing. In both reactors, lactate was the main initial product and was associated with growth of Streptococcus spp. (86% average relative abundance). Subsequently, lactate served as a substrate for secondary fermentations. In the reactor inoculated with rumen fluid from the Dutch cow, the relative abundance of Bacillus and Streptococcus increased from the start, and lactate, acetate, formate and ethanol were produced. From day 1.33 to 2, lactate and acetate were degraded, resulting in butyrate production. Butyrate production coincided with a decrease in relative abundance of Streptococcus spp. and increased relative abundances of bacteria of other groups, including Parabacteroides , Sporanaerobacter , Helicobacteraceae, Peptostreptococcaceae and Porphyromonadaceae. In the reactor with the Thai cow inoculum, Streptococcus spp. also increased from the start. When lactate was consumed, acetate, propionate and butyrate were produced (day 3-4). After day 3, bacteria belonging to five dominant groups, Bacteroides, Pseudoramibacter _ Eubacterium , Dysgonomonas , Enterobacteriaceae and Porphyromonadaceae, were detected and these showed significant positive correlations with acetate, propionate and butyrate levels. The complexity of rumen microorganisms with high adaptation capacity makes rumen fluid a suitable source to convert organic waste into valuable products without the addition of hydrolytic enzymes. Starch waste is a source for organic acid production, especially lactate.

Rumen prokaryotic communities of ruminants under different feeding paradigms on the Qinghai-Tibetan Plateau.

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Xue, Dan; Chen, Huai; Zhao, Xinquan; Xu, Shixiao; Hu, Linyong; Xu, Tianwei; Jiang, Lin; Zhan, Wei

2017-06-01

Yak and Tibetan sheep are the major indigenous ruminants on the Qinghai-Tibetan Plateau in China. The aim of this work was to study the differences in ruminal fermentation parameters and rumen prokaryotic community composition between hosts and feeding paradigms. The 16S rRNA genes targeting bacteria and archaea were sequenced using the MiSeq platform. The results showed that the prokaryotic community structure between yak and Tibetan sheep was significantly different (PTibetan sheep of the two groups (P=0.026). The core prokaryotic populations that existed in the rumen mostly dominated the structure. There was an obvious correlation of the prokaryotic community composition at the phylum and genus levels with the host or the feeding pattern. In addition, Tibetan sheep showed significantly higher yields of volatile fatty acids (VFAs) than yak, as did the NG group compared with the TMR group. In conclusion, both the host and feeding pattern may influence rumen microbial ecology system, with host effects being more important than those of the feeding pattern. Copyright © 2017 Elsevier GmbH. All rights reserved.

Associations of rumen parameters with feed efficiency and sampling routine in beef cattle.

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Lam, S; Munro, J C; Zhou, M; Guan, L L; Schenkel, F S; Steele, M A; Miller, S P; Montanholi, Y R

2017-11-10

Characterizing ruminal parameters in the context of sampling routine and feed efficiency is fundamental to understand the efficiency of feed utilization in the bovine. Therefore, we evaluated microbial and volatile fatty acid (VFA) profiles, rumen papillae epithelial and stratum corneum thickness and rumen pH (RpH) and temperature (RT) in feedlot cattle. In all, 48 cattle (32 steers plus 16 bulls), fed a high moisture corn and haylage-based ration, underwent a productive performance test to determine residual feed intake (RFI) using feed intake, growth, BW and composition traits. Rumen fluid was collected, then RpH and RT logger were inserted 5.5±1 days before slaughter. At slaughter, the logger was recovered and rumen fluid and rumen tissue were sampled. The relative daily time spent in specific RpH and RT ranges were determined. Polynomial regression analysis was used to characterize RpH and RT circadian patterns. Animals were divided into efficient and inefficient groups based on RFI to compare productive performance and ruminal parameters. Efficient animals consumed 1.8 kg/day less dry matter than inefficient cattle (P⩽0.05) while achieving the same productive performance (P⩾0.10). Ruminal bacteria population was higher (P⩽0.05) (7.6×1011 v. 4.3×1011 copy number of 16S rRNA gene/ml rumen fluid) and methanogen population was lower (P⩽0.05) (2.3×109 v. 4.9×109 copy number of 16S rRNA gene/ml rumen fluid) in efficient compared with inefficient cattle at slaughter with no differences (P⩾0.10) between samples collected on-farm. No differences (P⩾0.10) in rumen fluid VFA were also observed between feed efficiency groups either on-farm or at slaughter. However, increased (P⩽0.05) acetate, and decreased (P⩽0.05) propionate, butyrate, valerate and caproate concentrations were observed at slaughter compared with on-farm. Efficient had increased (P⩽0.05) rumen epithelium thickness (136 v. 126 µm) compared with inefficient cattle. Efficient animals

Kualitas Nutrisi Silase Berbahan Baku Singkong yang Diberi Enzim Cairan Rumen Sapi dan Leuconostoc mesenteroides

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S. Sandi

2010-04-01

Full Text Available The aim of this experiment was to evaluate the nutrient quality of cassava-based materials silage with cattle rumen liquor enzymes and Leuconostoc mesenteroides as poultry feed. The cassava material was hydrolyzed with cattle rumen liquor enzyme and incubated for 24 hours. The hydrolyzed product was added L. mesenteroides and ensiled in mini silo for 30 days. The experiment was designed in completely randomized design with 15 treatments and 3 replications. The result showed that temperature of cassava-based silage ranged from 26 to 30 oC. The flavor was sour and fresh fragrant and changed in color. Addition of cattle rumen liquor enzyme and L. mesenteroides bacteria significantly affected (P<0.05 pH (3.73-4.86, dry matter(30.14%-43.28%, cyanide (86.71%-96.50% and crude fiber content (0.78%-5.05%, but gave a fluctuate effect on protein content (-1.92%-2.39%. However, the treatment didnot affect dry matter losses (1.20%-2.66%. It is concluded that nutrient quality of cassava-based silage improved when it was added with cattle rumen liquor enzymes and L. mesenteroides by decreasing crude fiber and cyanide content. The best silage quality was obtained on tuber substrate and it increased protein KDUO (peel+leaves+tuber+tapioca waste silage.

Effect of sulfur supplements on cellulolytic rumen micro-organisms and microbial protein synthesis in cattle fed a high fibre diet.

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McSweeney, C S; Denman, S E

2007-11-01

To examine the effect of sulfur-containing compounds on the growth of anaerobic rumen fungi and the fibrolytic rumen bacteria Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes in pure culture and within the cattle rumen. The effect of two reduced sulfur compounds, 3-mercaptopropionic acid (MPA) or 3-mercapto-1-propanesulfonic acid as the sole S source on growth of pure fibroyltic fungal and bacterial cultures showed that these compounds were capable of sustaining growth. An in vivo trial was then conducted to determine the effect of sulfur supplements (MPA and sodium sulfate) on microbial population dynamics in cattle fed the roughage Dichanthium aristatum. Real-time PCR showed significant increases in fibrolytic bacterial and fungal populations when cattle were supplemented with these compounds. Sulfate supplementation leads to an increase in dry matter intake without a change in whole tract dry matter digestibility. Supplementation of low S-containing diets with either sodium sulfate or MPA stimulates microbial growth with an increase in rumen microbial protein supply to the animal. Through the use of real-time PCR monitoring, a better understanding of the effect of S supplementation on discrete microbial populations within the rumen is provided.

Effect of Supplementation of Branched Chain Fatty Acid on Colony of Ruminal Bacteria and Cell of Protozoa

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W Suryapratama

2009-05-01

Full Text Available A study was conducted to evaluate the potential of branched-chain volatile fatty acids (isobutyric, α-methylbutyric and β-methylbutiric that supplemented into the diet on the colony of ruminal bacteria and the cell of protozoa population. Five progeny Friesian Holstein males with initial weight 348±29 kg were used in a 5x5 Latin square design (30-d periods. The basal diet composed of 55% forage and 45% concentrate containing 10.5 MJ ME/kg and 15% crude protein (CP. There were five dietary treatments where A: basal diet, B: A+139 mg urea/kg W0.75, C: B+28 mg CaSO4/kg W0.75, D: C+0.05 mM isobutyric acid+0.05 mM β-methylbutyric acid, and E: D+0.05 mM α-methylbutyric acid. Rearing period was 30 days, consists of feed adaptation period 20 days, then growth observation was done within the last 10 days. Collection of ruminal fluid was done within the last day of observation period, and took 3-4 h after the feeding. The results showed that supplementation branched chain volatile fatty acids did not significant affect on the number of colonies of bacteria and protozoa population, but the significant effect (P<0.05 on the concentration of branched chain volatile fatty acids in the rumen fluid. The supplementation of α-methylbutyric (P <0.05 decreased of concentration of isobutyric and isovaleric in rumen fluid than the other treatments. It is concluded that supplementation of branched chain volatile fatty acids not used by rumen bacteria for their growth but for the elongation of fatty acid synthesis. The supplementation of branched chain volatile fatty acids was 0.05 mM not enough strong influence on the growth of colony of rumen bacteria. (Animal Production 11(2: 129-134 (2009 Key Words: rumen fermentation, branched-chain fatty acid, ruminal bacteria, protozoa

Study of the effect of presence or absence of protozoa on rumen fermentation and microbial protein contribution to the chyme.

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Belanche, A; Abecia, L; Holtrop, G; Guada, J A; Castrillo, C; de la Fuente, G; Balcells, J

2011-12-01

The aim of this study was to investigate the effect of presence or absence of protozoa on rumen fermentation and efficiency of microbial protein synthesis under different diets. Of 20 twin paired lambs, 1 lamb of each pair was isolated from the ewe within 24 h after birth and reared in a protozoa-free environment (n = 10), whereas their respective twin-siblings remained with the ewe (faunated, n = 10). When lambs reached 6 mo of age, 5 animals of each group were randomly allocated to 1 of 2 experimental diets consisting of either alfalfa hay as the sole diet, or 50:50 mixed with ground barley grain according to a 2 × 2 factorial arrangement of treatments. After 15 d of adaptation to the diet, the animals were euthanized and total rumen and abomasal contents were sampled to estimate rumen microbial synthesis using C(31) alkane as flow marker. Different ((15)N and purine bases) and a novel (recombinant DNA sequences) microbial markers, combined with several microbial reference extracts (rumen protozoa, liquid and solid associated bacteria) were evaluated. Absence of rumen protozoa modified the rumen fermentation pattern and decreased total tract OM and NDF digestibility in 2.0 and 5.1 percentage points, respectively. The effect of defaunation on microbial N flow was weak, however, and was dependent on the microbial marker and microbial reference extract considered. Faunated lambs fed with mixed diet showed the greatest rumen protozoal concentration and the least efficient microbial protein synthesis (29% less than the other treatments), whereas protozoa-free lambs fed with mixed diet presented the smallest ammonia concentration and 34% greater efficiency of N utilization than the other treatments. Although (15)N gave the most precise estimates of microbial synthesis, the use of recombinant DNA sequences represents an alternative that allows separate quantification of the bacteria and protozoa contributions. This marker showed that presence of protozoa decrease the

The effects of high dose of two manganese supplements (organic and inorganic) on the rumen microbial ecosystem.

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Kišidayová, Svetlana; Pristaš, Peter; Zimovčáková, Michaela; Blanár Wencelová, Monika; Homol'ová, Lucia; Mihaliková, Katarína; Čobanová, Klaudia; Grešáková, Ľubomíra; Váradyová, Zora

2018-01-01

Little is known about the effects of the high dose and types of manganese supplements on rumen environment at manganese intake level close above the limit of 150 mg/kg of dry feed matter. The effects of high dose of two manganese supplements (organic and inorganic) on rumen microbial ecosystem after four months of treatment of 18 lambs divided into three treatment groups were studied. We examined the enzyme activities (α-amylase, xylanase, and carboxymethyl cellulase), total and differential microscopic counts of rumen ciliates, total microscopic counts of bacteria, and fingerprinting pattern of the eubacterial and ciliates population analyzed by PCR-DGGE. Lambs were fed a basal diet with a basal Mn content (34.3 mg/kg dry matter; control) and supplemented either with inorganic manganous sulfate or organic Mn-chelate hydrate (daily 182.7, 184 mg/kg dry matter of feed, respectively). Basal diet, offered twice daily, consisted of ground barley and hay (268 and 732 g/kg dry matter per animal and day). The rumens of the lambs harbored ciliates of the genera of Entodinium, Epidinium, Diplodinium, Eudiplodinium, Dasytricha, and Isotricha. No significant differences between treatment groups were observed in the total ciliate number, the number of ciliates at the genus level, as well as the total number of bacteria. Organic Mn did decrease the species richness and diversity of the eubacterial population examined by PCR-DGGE. No effects of type of Mn supplement on the enzyme activities were observed. In comparison to the control, α-amylase specific activities were decreased and carboxymethyl-cellulase specific activities were increased by the Mn supplements. Xylanase activities were not influenced. In conclusion, our results suggested that the intake of tested inorganic and organic manganese supplements in excess may affect the specific groups of eubacteria. More studies on intake of Mn supplements at a level close to the limit can reveal if the changes in microbial

In vitro digestion of bloat-safe and bloat-causing legumes by rumen microorganisms: gas and foam production.

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Fay, J P; Cheng, K J; Hanna, M R; Howarth, R E; Costerton, J W

1980-08-01

Leaves of three bloat-safe legumes -- birdsfoot trefoil (Lotus corniculatus L.), sainfoin (Onobrychis viciaefolia Scop.), and cicer milkvetch (Astralagus cicer L.) -- and of three bloat-causing legumes -- alfalfa (Medicago sativa L.), red clover (Trifolium pratense L.), and white clover (Trifolium repens L.) -- were incubated with strained rumen fluid or with mixed rumen fluid and solids. Gas released was measured during the early period (0 to 22 h) of this in vitro digestion. Gas volume was greater with a 1:1 (wt/vol) mixture of solid and fluid rumen contents than with rumen fluid alone. It was greater with whole and chewed leaves from the bloat-causing legumes than with whole leaves from the bloat-safe legumes. However, when leaves were homogenized, volumes of gas from bloat-causing and bloat-safe legumes were similar. More gas was released from homogenized leaves than from the same weight of whole leaves. The amount of foam produced on chewed herbage and homogenized leaves of bloat-causing legumes was greater than on those of bloat-safe legumes. These results are consistent with the rate of disintegration and digestion of legumes by rumen bacteria being an important determinant in pasture bloat. Measurement of gas produced early in in vitro digestion may provide a useful bioassay for evaluating the bloat-causing potential of legumes in breeding selections if variability of the method can be reduced.

Biogas production from various coal types using beef cattle rumen's liquid as a source of microorganisms consortium

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Kurnani, Tubagus Benito Achmad; Harlia, Ellin; Hidayati, Yuli Astuti; Marlina, Eulis Tanti; Sugiarto, A. N.; Rahmah, K. N.; Joni, I. M.

2018-02-01

Nowadays, Indonesia is developing Coal-Bed Methane (CBM) production, but its production is not sufficient yet. Basically, CBM is produced naturally along with coal formation, i.e. through the activity of indigenous microorganisms. In this regard, to increase the production of CBM, adding a consortium of microorganisms into the coal deposit can be an option. One source of a consortium of bacteria available in nature is the rumen contents of ruminant livestock such as beef cattle. The purpose of this research was to know the capability of bacteria in rumen contents of beef cattle to produce CBM from various types of coal. In addition, to get a better concentration of bacteria than previous research so that it can be used as a reference for CBM production in the future. This explorative research used an experimental method with descriptive explanation. CBM production was performed using Hungate tube as a digester with three coal substrates, namely lignite, sub-bituminous and bituminous. This experiment also used 10-7 diluted rumen content of beef cattle as a source. The parameters measured were bacterial density, the amount of CBM, carbon dioxide and nitrous oxide on day 2, 5, 10 and 14. The treatment and parameters measurement were carried out in triplicates. This study finding showed that the highest bacterial density in all three types of coal was obtained on day 10 as well as the amount of CBM, carbon dioxide and nitrous oxide. These results are higher than the results from previous research therefore, this treatment can be used as an inoculant in a solid form for easy distribution.

The Genetic Diversity of Endophytic and Phyllosphere Bacteria from Several Indonesian Herbal Plants

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Devi Rachelia

2012-04-01

Full Text Available Herbal plants have been believed by Indonesians to be an alternative medicine to treat illnesses. The bioactivecompounds in the plant can be derived from secondary metabolites or from endophytic and phyllosphere bacteria whichcoexist within medicinal plants. A total of 18 endophytic bacteria and 32 phyllosphere bacteria were isolated from theherbal plants of Citrus sp., Pluchea indica, Curcuma longa, Nothopanax scuttelarium, Piper crocatum, andAndrographis paniculata. About 72% of endophytic bacteria isolates have proteolytic activity and about 11% havelipolytic activity. On the other hand, about 59% of phyllosphere bacteria isolates have proteolytic activity and about19% have lipolytic activity. Phylogenetic diversity analysis was conducted by using the amplified ribosomal DNArestriction analysis (ARDRA method and the sequence of 16S rDNA was digested with endonuclease restrictionenzymes: MspI, RsaI, and Sau961. The diversity of endophytic and phyllosphere bacterium from the samples of herbalplants was high. Bacteria isolated from the same herbal plant does not always have a close genetic relationship exceptfor the bacteria isolated from the P. indica plant which showed a close genetic relationship with each other.

Rumen and Cecum Microbiomes in Reindeer (Rangifer tarandus tarandus Are Changed in Response to a Lichen Diet and May Affect Enteric Methane Emissions.

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Alejandro Salgado-Flores

Full Text Available Reindeer (Rangifer tarandus tarandus are large Holarctic herbivores whose heterogeneous diet has led to the development of a unique gastrointestinal microbiota, essential for the digestion of arctic flora, which may include a large proportion of lichens during winter. Lichens are rich in plant secondary metabolites, which may affect members of the gut microbial consortium, such as the methane-producing methanogenic archaea. Little is known about the effect of lichen consumption on the rumen and cecum microbiotas and how this may affect methanogenesis in reindeer. Here, we examined the effects of dietary lichens on the reindeer gut microbiota, especially methanogens. Samples from the rumen and cecum were collected from two groups of reindeer, fed either lichens (Ld: n = 4, or a standard pelleted feed (Pd: n = 3. Microbial densities (methanogens, bacteria and protozoa were quantified using quantitative real-time PCR and methanogen and bacterial diversities were determined by 454 pyrosequencing of the 16S rRNA genes. In general, the density of methanogens were not significantly affected (p>0.05 by the intake of lichens. Methanobrevibacter constituted the main archaeal genus (>95% of reads, with Mbr. thaueri CW as the dominant species in both groups of reindeer. Bacteria belonging to the uncharacterized Ruminococcaceae and the genus Prevotella were the dominant phylotypes in the rumen and cecum, in both diets (ranging between 16-38% total sequences. Bacteria belonging to the genus Ruminococcus (3.5% to 0.6%; p = 0.001 and uncharacterized phylotypes within the order Bacteroidales (8.4% to 1.3%; p = 0.027, were significantly decreased in the rumen of lichen-fed reindeer, but not in the cecum (p = 0.2 and p = 0.087, respectively. UniFrac-based analyses showed archaeal and bacterial libraries were significantly different between diets, in both the cecum and the rumen (vegan::Adonis: pseudo-F<0.05. Based upon previous literature, we suggest that the

Metagenome Analysis of Protein Domain Collocation within Cellulase Genes of Goat Rumen Microbes

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SooYeon Lim

2013-08-01

Full Text Available In this study, protein domains with cellulase activity in goat rumen microbes were investigated using metagenomic and bioinformatic analyses. After the complete genome of goat rumen microbes was obtained using a shotgun sequencing method, 217,892,109 pair reads were filtered, including only those with 70% identity, 100-bp matches, and thresholds below E−10 using METAIDBA. These filtered contigs were assembled and annotated using blastN against the NCBI nucleotide database. As a result, a microbial community structure with 1431 species was analyzed, among which Prevotella ruminicola 23 bacteria and Butyrivibrio proteoclasticus B316 were the dominant groups. In parallel, 201 sequences related with cellulase activities (EC.3.2.1.4 were obtained through blast searches using the enzyme.dat file provided by the NCBI database. After translating the nucleotide sequence into a protein sequence using Interproscan, 28 protein domains with cellulase activity were identified using the HMMER package with threshold E values below 10−5. Cellulase activity protein domain profiling showed that the major protein domains such as lipase GDSL, cellulase, and Glyco hydro 10 were present in bacterial species with strong cellulase activities. Furthermore, correlation plots clearly displayed the strong positive correlation between some protein domain groups, which was indicative of microbial adaption in the goat rumen based on feeding habits. This is the first metagenomic analysis of cellulase activity protein domains using bioinformatics from the goat rumen.

Metagenomic analysis of rumen microbial population in dairy heifers fed a high grain diet supplemented with dicarboxylic acids or polyphenols.

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De Nardi, Roberta; Marchesini, Giorgio; Li, Shucong; Khafipour, Ehsan; Plaizier, Kees J C; Gianesella, Matteo; Ricci, Rebecca; Andrighetto, Igino; Segato, Severino

2016-02-19

The aim of this study was to investigate the effects of two feed supplements on rumen bacterial communities of heifers fed a high grain diet. Six Holstein-Friesian heifers received one of the following dietary treatments according to a Latin square design: no supplement (control, C), 60 g/day of fumarate-malate (organic acid, O) and 100 g/day of polyphenol-essential oil (P). Rumen fluid was analyzed to assess the microbial population using Illumina sequencing and quantitative real time PCR. The P treatment had the highest number of observed species (P PCoA with unweighted Unifrac distance showed a separation among dietary treatments (P = 0.09), above all between the C and P (P = 0.05). The O and P treatments showed a significant increase of the family Christenenellaceae and a decline of Prevotella brevis compared to C. Additionally, the P treatment enhanced the abundance of many taxa belonging to Bacteroidetes, Firmicutes and Tenericutes phyla due to a potential antimicrobial activity of flavonoids that increased competition among bacteria. Organic acid and polyphenols significantly modified rumen bacterial populations during high-grain feeding in dairy heifers. In particular the polyphenol treatment increased the richness and diversity of rumen microbiota, which are usually high in conditions of physiological rumen pH and rumen function.

Metagenomic analysis of buffalo rumen microbiome: Effect of roughage diet on Dormancy and Sporulation genes.

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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.

Feeding, evaluating, and controlling rumen function.

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Lean, Ian J; Golder, Helen M; Hall, Mary Beth

2014-11-01

Achieving optimal rumen function requires an understanding of feeds and systems of nutritional evaluation. Key influences on optimal function include achieving good dry matter intake. The function of feeds in the rumen depends on other factors including chemical composition, rate of passage, degradation rate of the feed, availability of other substrates and cofactors, and individual animal variation. This article discusses carbohydrate, protein, and fat metabolism in the rumen, and provides practical means of evaluation of rations in the field. Conditions under which rumen function is suboptimal (ie, acidosis and bloat) are discussed, and methods for control examined. Copyright © 2014 Elsevier Inc. All rights reserved.

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

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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

Characterization of rumen bacterial diversity and fermentation parameters in concentrate fed cattle with and without forage.

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Petri, R M; Forster, R J; Yang, W; McKinnon, J J; McAllister, T A

2012-06-01

To determine the effects of the removal of forage in high-concentrate diets on rumen fermentation conditions and rumen bacterial populations using culture-independent methods. Detectable bacteria and fermentation parameters were measured in the solid and liquid fractions of digesta from cattle fed two dietary treatments, high concentrate (HC) and high concentrate without forage (HCNF). Comparison of rumen fermentation conditions showed that duration of time spent below pH 5·2 and rumen osmolality were higher in the HCNF treatment. Simpson's index of 16S PCR-DGGE images showed a greater diversity of dominant species in the HCNF treatment. Real-time qPCR showed populations of Fibrobacter succinogenes (P = 0·01) were lower in HCNF than HC diets. Ruminococcus spp., F. succinogenes and Selenomonas ruminantium were at higher (P ≤ 0·05) concentrations in the solid vs the liquid fraction of digesta regardless of diet. The detectable bacterial community structure in the rumen is highly diverse. Reducing diet complexity by removing forage increased bacterial diversity despite the associated reduction in ruminal pH being less conducive for fibrolytic bacterial populations. Quantitative PCR showed that removal of forage from the diet resulted in a decline in the density of some, but not all fibrolytic bacterial species examined. Molecular techniques such as DGGE and qPCR provide an increased understanding of the impacts of dietary changes on the nature of rumen bacterial populations, and conclusions derived using these techniques may not match those previously derived using traditional laboratory culturing techniques. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

Impact of levels of total digestible nutrients on microbiome, enzyme profile and degradation of feeds in buffalo rumen.

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Kala, Anju; Kamra, D N; Kumar, Avinash; Agarwal, Neeta; Chaudhary, L C; Joshi, C G

2017-01-01

The present study was aimed at understanding a shift in rumen microbiome of buffaloes fed various levels of total digestible nutrients. To understand the process, the metagenomics of rumen microbes, in vivo and in vitro rumen fermentation studies were carried out. Three rumen fistulated adult male Murrah buffaloes were fed three isonitrogenous diets varying in total digestible nutrients (70, 85 and 100% of TDN requirement) in 3X3 switch over design. On dry matter basis, wheat straw/ roughage content were 81, 63 and 51% and that of maize grain was 8, 16 and 21% in three diets respectively. After 20 d of feeding, rumen liquor and rumen contents were sampled just before (0h) and 4h post feeding. Ruminococcus flavefaciens and R. albus (estimated with real time PCR) were higher in high roughage diets. The predominant phyla in all the three groups were Bacteroidetes, Firmicutes followed by Proteobacteria, Actinobacteria and Fibrobacteres. A core group of more than fifty rumen bacteria was present in all the animals with very little variations due to level of TDN. The most predominant bacterial genera reported in order of decreasing abundance were: Prevotella, Bacteroides, Clostridium, Ruminococcus, Eubacterium, Parabacteroides, Fibrobacter, Butyrivibrio etc. The higher diversity of the enyzmes families GH 23, GH 28, GH 39, GH 97, GH 106, and GH 127 (the enzymes active in fibre and starch degradation) were significantly higher on 100%TDN diet while CE 14 (required for the hydrolysis of bond between carbohydrate and lignin) was higher on low TDN (70%) diet, indicating ester bond cleavage was better in animals fed high roughage (wheat straw) diet.

Sekuensing 16S DNA Bakteri Selulolitik Asal Limbah Cairan Rumen Sapi Peranakan Ongole (SEQUENCING OF 16S DNA OF CELLULOLYTIC BACTERIA FROM BOVINE RUMEN FLUID WASTE ONGOLE CROSSBREED

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Widya Paramita Lokapirnasari

2017-04-01

Full Text Available This study aimed to identified cellulolytic inoculant code WPL 214 isolated from bovine rumen fluid waste of Ongole Cross Breed of Surabaya Slaughter house. A single colony of isolates celulolytic grown on 5 mL of liquid media Luria Bertani (LB consist of 1 % NaCl , 1% tripton , 0.5 % yeast extract, containing1 % carboxymethyl cellulose (CMC at temperature 37°C, using a shaker of incubator during 16-18 hours. That isolate determined by 16S DNA gen analysis using High Fidelity Platinum Taq DNA Polymerase with primer forward PB36 5’-AGR GTT TGA TCM TGG CTC AG-3’ and primer reverse PB38 5’-GMT ACCTTG TTA CGA CTT-3’ for PCR. Nucleotide sequence of 16S DNA fragment was determined through the sequencing method. The result was then compared with GenBank database to recognize the type of the sample bacteria. DNA isolation and 16S DNA coding genes amplification were carried out using Kit High Fidelity Platinum Taq DNA Polymerase. Afterward, BLAST was applied to identify the phylogenetic tree. The bacteria was capable of indicating the existence of clear zone in a media CMC by congo red staining. The existence of the clear zone associated with the activity of microbes to degrade cellulose. The conclusión of this research based on the results was the sequencing nucleotides genome 16S DNA showed that cellulolytic inoculant was identified as Enterobacter cloacae WPL 214. ABSTRAK Penelitian ini bertujuan untuk mengidentifikasi lebih lanjut isolat selulolitik kode WPL 214 yang telah diisolasi dari cairan rumen sapi peranakan ongole dari limbah Rumah Potong Hewan Surabaya. Koloni tunggal dari isolat selulolitik ditumbuhkan pada 5 mL media cair Luria Bertani (LB dengan komposisisi 1% NaCl, 1% tripton, 0,5% yeast ekstrak, yang mengandung 1% substrat carboxymethyl cellulose (CMC pada suhu 37°C, dengan pengocokan menggunakan shaker incubator selama ±16-18 jam. Penelitian ini terdiri dari dua tahap, tahap pertama dilakukan isolasi DNA, tahap kedua

Associative patterns among anaerobic fungi, methanogenic archaea and bacterial communities in response to changes in diet and age in the rumen of dairy cows

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Sanjay eKumar

2015-07-01

Full Text Available 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

Gene and transcript abundances of bacterial type III secretion systems from the rumen microbiome are correlated with methane yield in sheep.

Science.gov (United States)

Kamke, Janine; Soni, Priya; Li, Yang; Ganesh, Siva; Kelly, William J; Leahy, Sinead C; Shi, Weibing; Froula, Jeff; Rubin, Edward M; Attwood, Graeme T

2017-08-08

Ruminants are important contributors to global methane emissions via microbial fermentation in their reticulo-rumens. This study is part of a larger program, characterising the rumen microbiomes of sheep which vary naturally in methane yield (g CH 4 /kg DM/day) and aims to define differences in microbial communities, and in gene and transcript abundances that can explain the animal methane phenotype. Rumen microbiome metagenomic and metatranscriptomic data were analysed by Gene Set Enrichment, sparse partial least squares regression and the Wilcoxon Rank Sum test to estimate correlations between specific KEGG bacterial pathways/genes and high methane yield in sheep. KEGG genes enriched in high methane yield sheep were reassembled from raw reads and existing contigs and analysed by MEGAN to predict their phylogenetic origin. Protein coding sequences from Succinivibrio dextrinosolvens strains were analysed using Effective DB to predict bacterial type III secreted proteins. The effect of S. dextrinosolvens strain H5 growth on methane formation by rumen methanogens was explored using co-cultures. Detailed analysis of the rumen microbiomes of high methane yield sheep shows that gene and transcript abundances of bacterial type III secretion system genes are positively correlated with methane yield in sheep. Most of the bacterial type III secretion system genes could not be assigned to a particular bacterial group, but several genes were affiliated with the genus Succinivibrio, and searches of bacterial genome sequences found that strains of S. dextrinosolvens were part of a small group of rumen bacteria that encode this type of secretion system. In co-culture experiments, S. dextrinosolvens strain H5 showed a growth-enhancing effect on a methanogen belonging to the order Methanomassiliicoccales, and inhibition of a representative of the Methanobrevibacter gottschalkii clade. This is the first report of bacterial type III secretion system genes being associated with high

FibroChip, a Functional DNA Microarray to Monitor Cellulolytic and Hemicellulolytic Activities of Rumen Microbiota

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Sophie Comtet-Marre

2018-02-01

Full Text Available Ruminants fulfill their energy needs for growth primarily through microbial breakdown of plant biomass in the rumen. Several biotic and abiotic factors influence the efficiency of fiber degradation, which can ultimately impact animal productivity and health. To provide more insight into mechanisms involved in the modulation of fibrolytic activity, a functional DNA microarray targeting genes encoding key enzymes involved in cellulose and hemicellulose degradation by rumen microbiota was designed. Eight carbohydrate-active enzyme (CAZyme families (GH5, GH9, GH10, GH11, GH43, GH48, CE1, and CE6 were selected which represented 392 genes from bacteria, protozoa, and fungi. The DNA microarray, designated as FibroChip, was validated using targets of increasing complexity and demonstrated sensitivity and specificity. In addition, FibroChip was evaluated for its explorative and semi-quantitative potential. Differential expression of CAZyme genes was evidenced in the rumen bacterium Fibrobacter succinogenes S85 grown on wheat straw or cellobiose. FibroChip was used to identify the expressed CAZyme genes from the targeted families in the rumen of a cow fed a mixed diet based on grass silage. Among expressed genes, those encoding GH43, GH5, and GH10 families were the most represented. Most of the F. succinogenes genes detected by the FibroChip were also detected following RNA-seq analysis of RNA transcripts obtained from the rumen fluid sample. Use of the FibroChip also indicated that transcripts of fiber degrading enzymes derived from eukaryotes (protozoa and anaerobic fungi represented a significant proportion of the total microbial mRNA pool. FibroChip represents a reliable and high-throughput tool that enables researchers to monitor active members of fiber degradation in the rumen.

FibroChip, a Functional DNA Microarray to Monitor Cellulolytic and Hemicellulolytic Activities of Rumen Microbiota.

Science.gov (United States)

Comtet-Marre, Sophie; Chaucheyras-Durand, Frédérique; Bouzid, Ourdia; Mosoni, Pascale; Bayat, Ali R; Peyret, Pierre; Forano, Evelyne

2018-01-01

Ruminants fulfill their energy needs for growth primarily through microbial breakdown of plant biomass in the rumen. Several biotic and abiotic factors influence the efficiency of fiber degradation, which can ultimately impact animal productivity and health. To provide more insight into mechanisms involved in the modulation of fibrolytic activity, a functional DNA microarray targeting genes encoding key enzymes involved in cellulose and hemicellulose degradation by rumen microbiota was designed. Eight carbohydrate-active enzyme (CAZyme) families (GH5, GH9, GH10, GH11, GH43, GH48, CE1, and CE6) were selected which represented 392 genes from bacteria, protozoa, and fungi. The DNA microarray, designated as FibroChip, was validated using targets of increasing complexity and demonstrated sensitivity and specificity. In addition, FibroChip was evaluated for its explorative and semi-quantitative potential. Differential expression of CAZyme genes was evidenced in the rumen bacterium Fibrobacter succinogenes S85 grown on wheat straw or cellobiose. FibroChip was used to identify the expressed CAZyme genes from the targeted families in the rumen of a cow fed a mixed diet based on grass silage. Among expressed genes, those encoding GH43, GH5, and GH10 families were the most represented. Most of the F. succinogenes genes detected by the FibroChip were also detected following RNA-seq analysis of RNA transcripts obtained from the rumen fluid sample. Use of the FibroChip also indicated that transcripts of fiber degrading enzymes derived from eukaryotes (protozoa and anaerobic fungi) represented a significant proportion of the total microbial mRNA pool. FibroChip represents a reliable and high-throughput tool that enables researchers to monitor active members of fiber degradation in the rumen.

Fractional rate of degradation (kd) of starch in the rumen and its relation to in vivo rumen and total digestibility

DEFF Research Database (Denmark)

Hvelplund, Torben; Larsen, Mogens; Lund, Peter

2009-01-01

in different ways both chemically and physically. The starch sources were fed in mixed diets together with grass silage and soya bean meal and allocated ad libitum to fistulated dairy cows. The starch content varied between 13 and 35% in ration dry matter for the different starch sources. The design...... was a series of cross-over experiments with two cows and two periods. Ruminal starch pool was estimated from rumen evacuation and starch flow was estimated by duodenal and faeces sampling. Fractional rate of rumen degradation was estimated from the equation [kd = rumen degraded/rumen pool] and rumen and total...

In Vitro Rumen Fermentation Characteristics and Fatty Acid Profiles Added with Calcium Soap of Canola/Flaxseed Oil

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S. Suharti

2017-12-01

Full Text Available This research aimed to assess the effect of adding canola oil and flaxseed oil which were protected with calcium soap (Ca-soap on the fermentation characteristics, rumen microbial population, and the profile of fatty acids in the rumen during 4 and 8 hours in the in vitro fermentation. The research design used in this study was a completely randomized block design with 3 treatments and 4 replications. The treatments consisted of control ration (Napier grass and concentrate at the ratio of 60 : 40, control + 6% of Ca-soap of canola oil, and control + 6% of Ca-soap of flaxseed oil. Variables observed were pH value, NH3 concentration, volatile fatty acid (VFA, dry matter and organic matter digestibility, and fatty acid profile.  The results showed that the addition of Ca-soap of canola or flaxseed oil did not affect the pH value, NH3 concentration, dry matter digestibility, organic matter digestibility, total population of bacteria and protozoa in the rumen. However, the total production of ruminal VFA was increased (P<0.05 with the addition of Ca soap of canola oil/flaxseed oil. The use of Ca-soap of flaxseed oil increased (P<0.05 the content of unsaturated fatty acids in the rumen at 4 h incubation. The addition of Ca-soap of flaxseed oil resulted the lowest (P<0.05 level of unsaturated fatty acids biohydrogenation compared to the other treatments at 4 h incubation. In conclusion, the addition of Ca soap of canola/flaxseed oil could improve VFA total production. Vegetable oils protected using calcium soap could inhibit unsaturated fatty acid biohidrogenation by rumen microbes. Ca-soap of flaxseed oil could survive from rumen biohydrogenation in the rumen better than Ca-soap of canola oil.

Repeated inoculation of cattle rumen with bison rumen contents alters the rumen microbiome and improves nitrogen digestibility in cattle

OpenAIRE

Ribeiro, Gabriel O.; Oss, Daniela B.; He, Zhixiong; Gruninger, Robert J.; Elekwachi, Chijioke; Forster, Robert J.; Yang, WenZhu; Beauchemin, Karen A.; McAllister, Tim A.

2017-01-01

Future growth in demand for meat and milk, and the socioeconomic and environmental challenges that farmers face, represent a ?grand challenge for humanity?. Improving the digestibility of crop residues such as straw could enhance the sustainability of ruminant production systems. Here, we investigated if transfer of rumen contents from bison to cattle could alter the rumen microbiome and enhance total tract digestibility of a barley straw-based diet. Beef heifers were adapted to the diet for ...

Functional gene profiling through metaRNAseq approach reveals diet-dependent variation in rumen microbiota of buffalo (Bubalus bubalis).

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Hinsu, Ankit T; Parmar, Nidhi R; Nathani, Neelam M; Pandit, Ramesh J; Patel, Anand B; Patel, Amrutlal K; Joshi, Chaitanya G

2017-04-01

Recent advances in next generation sequencing technology have enabled analysis of complex microbial community from genome to transcriptome level. In the present study, metatranscriptomic approach was applied to elucidate functionally active bacteria and their biological processes in rumen of buffalo (Bubalus bubalis) adapted to different dietary treatments. Buffaloes were adapted to a diet containing 50:50, 75:25 and 100:0 forage to concentrate ratio, each for 6 weeks, before ruminal content sample collection. Metatranscriptomes from rumen fiber adherent and fiber-free active bacteria were sequenced using Ion Torrent PGM platform followed by annotation using MG-RAST server and CAZYmes (Carbohydrate active enzymes) analysis toolkit. In all the samples Bacteroidetes was the most abundant phylum followed by Firmicutes. Functional analysis using KEGG Orthology database revealed Metabolism as the most abundant category at level 1 within which Carbohydrate metabolism was dominating. Diet treatments also exerted significant differences in proportion of enzymes involved in metabolic pathways for VFA production. Carbohydrate Active Enzyme(CAZy) analysis revealed the abundance of genes encoding glycoside hydrolases with the highest representation of GH13 CAZy family in all the samples. The findings provide an overview of the activities occurring in the rumen as well as active bacterial population and the changes occurring through different dietary treatments. Copyright © 2017 Elsevier Ltd. All rights reserved.

Metagenomic insights into lignocellulose-degrading genes through Illumina-based de novo sequencing of the microbiome in Vietnamese native goats' rumen

NARCIS (Netherlands)

Do, Thi Huyen; Le, Ngoc Giang; Dao, Trong Khoa; Nguyen, Thi Mai Phuong; Le, Tung Lam; Luu, Han Ly; Nguyen, Khanh Hoang Viet; Nguyen, Van Lam; Le, Lan Anh; Phung, Thu Nguyet; van Straalen, Nico M; Roelofs, Dick; Truong, Nam Hai

2018-01-01

The scarcity of enzymes having an optimal activity in lignocellulose deconstruction is an obstacle for industrial-scale conversion of cellulosic biomass into biofuels. With the aim of mining novel lignocellulolytic enzymes, a ~9 Gb metagenome of bacteria in Vietnamese native goats' rumen was

Fatty acid composition and bacterial community changes in the rumen fluid of lactating sheep fed sunflower oil plus incremental levels of marine algae.

Science.gov (United States)

Toral, P G; Belenguer, A; Shingfield, K J; Hervás, G; Toivonen, V; Frutos, P

2012-02-01

Supplementation of ruminant diets with plant oils and marine lipids is an effective strategy for lowering saturated fatty acid (FA) content and increasing the concentration of cis-9,trans-11 conjugated linoleic acid and long-chain n-3 FA in ruminant milk. However, changes in populations of ruminal microorganisms associated with altered biohydrogenation of dietary unsaturated FA are not well characterized. Twenty-five lactating Assaf ewes were allocated at random to 1 of 5 treatments composed of dehydrated alfalfa hay and concentrates containing no additional lipid (control), or supplemented with 25 g of sunflower oil and 0 (SO), 8 (SOMA(1)), 16 (SOMA(2)), or 24 (SOMA(3)) g of marine algae/kg of diet dry matter. On d 28 on diet, samples of rumen fluid were collected for lipid analysis and microbial DNA extraction. Appearance and identification of biohydrogenation intermediates was determined based on complementary gas chromatography and Ag+-HPLC analysis of FA methyl esters. Total bacteria and the Butyrivibrio group were studied in microbial DNA by terminal RFLP analysis, and real-time PCR was used to quantify the known Butyrivibrio bacteria that produce trans-11 18:1 or 18:0. Dietary supplements of sunflower oil alone or in combination with marine algae altered the FA profile of rumen fluid, which was associated with changes in populations of specific bacteria. Inclusion of marine algae in diets containing sunflower oil resulted in the accumulation of trans 18:1 and 10-O-18:0 and a marked decrease in 18:0 concentrations in rumen fluid. At the highest levels of supplementation (SOMA(2) and SOMA(3)), marine algae also promoted a shift in ruminal biohydrogenation pathways toward the formation of trans-10 18:1 at the expense of trans-11 18:1. Changes in the concentration of biohydrogenation intermediates were not accompanied by significant variations in the abundance of known cultivated ruminal bacteria capable of hydrogenating unsaturated FA. However, certain

Enhanced biogas yield from energy crops with rumen anaerobic fungi

Energy Technology Data Exchange (ETDEWEB)

Prochazka, Jindrich; Zabranska, Jana; Dohanyos, Michal [Department of Water Technology and Environmental Engineering, Faculty of Environmental Technology, Institute of Chemical Technology in Prague, Prague (Czech Republic); Mrazek, Jakub; Strosova, Lenka; Fliegerova, Katerina [Laboratory of Anaerobic Microbiology, Institute of Animal Physiology and Genetics, CAS, v.v.i., Prague (Czech Republic)

2012-06-15

Anaerobic fungi (AF) are able to degrade crop substrates with higher efficiency than commonly used anaerobic bacteria. The aim of this study was to investigate ways of use of rumen AF to improve biogas production from energy crops under laboratory conditions. In this study, strains of AF isolated from feces or rumen fluid of cows and deer were tested for their ability to integrate into the anaerobic bacterial ecosystem used for biogas production, in order to improve degradation of substrate polysaccharides and consequently the biogas yield. Batch culture, fed batch culture, and semicontinuous experiments have been performed using anaerobic sludge from pig slurry fermentation and different kinds of substrates (celluloses, maize, and grass silage) inoculated by different genera of AF. All experiments showed a positive effect of AF on the biogas yield and quality. AF improved the biogas production by 4-22%, depending on the substrate and AF species used. However, all the cultivation experiments indicated that rumen fungi do not show long-term survival in fermenters with digestate from pig slurry. The best results were achieved during fed batch experiment with fungal culture Anaeromyces (KF8), in which biogas production was enhanced during the whole experimental period of 140 days. This result has not been achieved in semicontinuous experiment, where increment in biogas production in fungal enriched reactor was only 4% after 42 days. (copyright 2012 Wiley-VCH Verlag GmbH and Co. KGaA, Weinheim)

The proteolytic system of Lactococcus lactis

NARCIS (Netherlands)

Kunji, Edmundus Richardus Stephanus

1997-01-01

The bacterium Lactococcus lactis usues an extencive proteolytic system to utilize milk proteins (caseins) in orde to meet its need for amino acids. The genetic and biochemical properties of the putative components of the proteolytic pathway are well-described. However, little is known about the role

Rumen content stratification in the giraffe (Giraffa camelopardalis).

Science.gov (United States)

Sauer, Cathrine; Clauss, Marcus; Bertelsen, Mads F; Weisbjerg, Martin R; Lund, Peter

2017-01-01

Ruminants differ in the degree of rumen content stratification, with 'cattle-types' (i.e., the grazing and intermediate feeding ruminants) having stratified content, whereas 'moose-types' (i.e., the browsing ruminants) have unstratified content. The feeding ecology, as well as the digestive morphophysiology of the giraffe (Giraffa camelopardalis), suggest that it is a 'moose-type' ruminant. Correspondingly, the giraffe should have an unstratified rumen content and an even rumen papillation pattern. Digesta samples were collected from along the digestive tract of 27 wild-caught giraffes kept in bomas for up to 2months, and 10 giraffes kept in zoological gardens throughout their lives. Samples were analysed for concentration of dry matter, fibre fractions, volatile fatty acids and NH 3 , as well as mean particle size and pH. There was no difference between the dorsal and ventral rumen region in any of these parameters, indicating homogenous rumen content in the giraffes. In addition to the digesta samples, samples of dorsal rumen, ventral rumen and atrium ruminis mucosa were collected and the papillary surface enlargement factor was determined, as a proxy for content stratification. The even rumen papillation pattern observed also supported the concept of an unstratified rumen content in giraffes. Zoo giraffes had a slightly more uneven papillation pattern than boma giraffes. This finding could not be matched by differences in physical characteristics of the rumen content, probably due to an influence of fasting time ante mortem on these parameters. Copyright © 2016 Elsevier Inc. All rights reserved.

Ultrasonographic Examination of the Rumen in Healthy Cows

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Sheikh Imran

2011-01-01

Full Text Available 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.

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

Science.gov (United States)

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

2015-02-01

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. 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. Thyme oil at higher dose level (600 ppm) reduced (p0.05) in 300 and 600 ppm dose levels. 600 ppm dose level of clove oil reduced (pclove and peppermint oil. Right combination of these essential oils may prove to enhance performance of animals by reducing methane production and inhibiting protein degradation in rumen.

The effect of dietary Chlorella vulgaris supplementation on micro-organism community, enzyme activities and fatty acid profile in the rumen liquid of goats.

Science.gov (United States)

Tsiplakou, E; Abdullah, M A M; Skliros, D; Chatzikonstantinou, M; Flemetakis, E; Labrou, N; Zervas, G

2017-04-01

Microalgae might be considered as an alternative source of fat and/or protein for ruminant's diets. However, changes in populations of ruminal micro-organisms associated with biohydrogenation process, methane and ammonia production in response to microalgae dietary supplementation have not been well characterized. Thus, 16 cross-bred goats were divided into two groups. Each goat of both groups was fed individually with alfalfa hay and concentrates separately. The concentrates of the control group had no microalgae while those of the treated group were supplemented with 10 g lyophilized Chlorella vulgaris/kg concentrate (chlor). On the 30th experimental day, samples of rumen fluid were collected for microbial DNA extraction, fatty acid profile and enzyme activity analyses. The results showed that the chlor diet compared with the control increased significantly the populations of Methanosphaera stadtmanae, Methanobrevibacter ruminantium and Methanogens bacteria and protozoa in the rumen of goats. A significant reduction in the cellulase activity and in the abundance of Ruminococcus albus, and a significant increase in the protease activity and in the abundance of Clostridium sticklandii in the rumen liquid of goats fed with the chlor diet, compared with the control, were found. Chlorella vulgaris supplementation promoted the formation of trans C 18:1 , trans-11 C 18:1 and monounsaturated fatty acids (MUFA), while the proportions of C 18:0 and long-chain fatty acids (LCFA) reduced significantly in the rumen liquid of goats. This shift in ruminal biohydrogenation pathway was accompanied by a significant increase in Butyrivibrio fibrisolvens trans C 18:1 -producing bacteria. In conclusion, the supplementation of diets with microalgae needs further investigation because it enhances the populations of methane-producing bacteria and protozoa. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

The Role of Ciliate Protozoa in the Rumen.

Science.gov (United States)

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.

Diversity of archaea and bacteria in a biogas reactor fed with ...

African Journals Online (AJOL)

Diversity of archaea and bacteria in a biogas reactor fed with Pennisetum sinese ... AFRICAN JOURNALS ONLINE (AJOL) · Journals · Advanced Search · USING ... rumen microbial community in a biogas reactor by 16S rRNA gene analysis, ...

Illumina Sequencing Approach to Characterize Thiamine Metabolism Related Bacteria and the Impacts of Thiamine Supplementation on Ruminal Microbiota in Dairy Cows Fed High-Grain Diets.

Science.gov (United States)

Pan, Xiaohua; Xue, Fuguang; Nan, Xuemei; Tang, Zhiwen; Wang, Kun; Beckers, Yves; Jiang, Linshu; Xiong, Benhai

2017-01-01

The requirements of thiamine in adult ruminants are mainly met by ruminal bacterial synthesis, and thiamine deficiencies will occur when dairy cows overfed with high grain diet. However, there is limited knowledge with regard to the ruminal thiamine synthesis bacteria, and whether thiamine deficiency is related to the altered bacterial community by high grain diet is still unclear. To explore thiamine synthesis bacteria and the response of ruminal microbiota to high grain feeding and thiamine supplementation, six rumen-cannulated Holstein cows were randomly assigned into a replicated 3 × 3 Latin square design trial. Three treatments were control diet (CON, 20% dietary starch, DM basis), high grain diet (HG, 33.2% dietary starch, DM basis) and high grain diet supplemented with 180 mg thiamine/kg DMI (HG+T). On day 21 of each period, rumen content samples were collected at 3 h postfeeding. Ruminal thiamine concentration was detected by high performance liquid chromatography. The microbiota composition was determined using Illumina MiSeq sequencing of 16S rRNA gene. Cows receiving thiamine supplementation had greater ruminal pH value, acetate and thiamine content in the rumen. Principal coordinate analysis and similarity analysis indicated that HG feeding and thiamine supplementation caused a strong shift in bacterial composition and structure in the rumen. At the genus level, compared with CON group, the relative abundances of 19 genera were significantly changed by HG feeding. Thiamine supplementation increased the abundance of cellulolytic bacteria including Bacteroides, Ruminococcus 1, Pyramidobacter, Succinivibrio , and Ruminobacter , and their increases enhanced the fiber degradation and ruminal acetate production in HG+T group. Christensenellaceae R7, Lachnospira, Succiniclasticum , and Ruminococcaceae NK4A214 exhibited a negative response to thiamine supplementation. Moreover, correlation analysis revealed that ruminal thiamine concentration was positively

Illumina Sequencing Approach to Characterize Thiamine Metabolism Related Bacteria and the Impacts of Thiamine Supplementation on Ruminal Microbiota in Dairy Cows Fed High-Grain Diets

Directory of Open Access Journals (Sweden)

Xiaohua Pan

2017-09-01

Full Text Available The requirements of thiamine in adult ruminants are mainly met by ruminal bacterial synthesis, and thiamine deficiencies will occur when dairy cows overfed with high grain diet. However, there is limited knowledge with regard to the ruminal thiamine synthesis bacteria, and whether thiamine deficiency is related to the altered bacterial community by high grain diet is still unclear. To explore thiamine synthesis bacteria and the response of ruminal microbiota to high grain feeding and thiamine supplementation, six rumen-cannulated Holstein cows were randomly assigned into a replicated 3 × 3 Latin square design trial. Three treatments were control diet (CON, 20% dietary starch, DM basis, high grain diet (HG, 33.2% dietary starch, DM basis and high grain diet supplemented with 180 mg thiamine/kg DMI (HG+T. On day 21 of each period, rumen content samples were collected at 3 h postfeeding. Ruminal thiamine concentration was detected by high performance liquid chromatography. The microbiota composition was determined using Illumina MiSeq sequencing of 16S rRNA gene. Cows receiving thiamine supplementation had greater ruminal pH value, acetate and thiamine content in the rumen. Principal coordinate analysis and similarity analysis indicated that HG feeding and thiamine supplementation caused a strong shift in bacterial composition and structure in the rumen. At the genus level, compared with CON group, the relative abundances of 19 genera were significantly changed by HG feeding. Thiamine supplementation increased the abundance of cellulolytic bacteria including Bacteroides, Ruminococcus 1, Pyramidobacter, Succinivibrio, and Ruminobacter, and their increases enhanced the fiber degradation and ruminal acetate production in HG+T group. Christensenellaceae R7, Lachnospira, Succiniclasticum, and Ruminococcaceae NK4A214 exhibited a negative response to thiamine supplementation. Moreover, correlation analysis revealed that ruminal thiamine concentration was

Selection of proteolytic bacteria with ability to inhibit Vibrio harveyi during white shrimp (Litopenaeus vannamei cultivation

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Suntinanalert, P.

2007-03-01

Full Text Available Five isolates of bacteria with high proteolytic activity, isolated from water samples of intensive shrimp ponds in southern Thailand, were selected to test for the ability to control the shrimp pathogen Vibrioharveyi. 70 μl of each culture broth were investigated for their ability to inhibit V. harveyi using an agar well diffusion test but only one isolate W3 gave a reasonable sized inhibition zone of 21.62 mm. This zone wassimilar to that of oxolinic acid (2 μg and sulfamethoxazole (25 μg. The W3 isolate was identified as Pseudomonas sp. Shrimp cultivation in aquaria was conducted to investigate the inhibition of V. harveyi bythe isolate W3. The experiment consisted of a treatment of the shrimp culture with an inoculum of the isolate W3 and V. harveyi (biocontrol set, a positive control set (only inoculation of V. harveyi and a negativecontrol set as without inoculation. No mortality was found in the negative control. Shrimp mortality in the biocontrol set (33% was lower than that in the positive control set (40%; however, it showed no significantdifference (p>0.05. The average numbers of V. harveyi over 12 days of the biocontrol set were lower than those in the positive control set by about 1 log cycle although the numbers were not significantly different(p>0.05. The shrimp growth rate at day 32 of cultivation was in order of the biocontrol treatment (10.17% > the negative control treatment (9.44% > the positive control set (9.28%, but no significant difference (p>0.05 was observed among treatments.

Total rRNA-Seq Analysis Gives Insight into Bacterial, Fungal, Protozoal and Archaeal Communities in the Rumen Using an Optimized RNA Isolation Method

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Chijioke O. Elekwachi

2017-09-01

Full Text Available Advances in high throughput, next generation sequencing technologies have allowed an in-depth examination of biological environments and phenomena, and are particularly useful for culture-independent microbial community studies. Recently the use of RNA for metatranscriptomic studies has been used to elucidate the role of active microbes in the environment. Extraction of RNA of appropriate quality is critical in these experiments and TRIzol reagent is often used for maintaining stability of RNA molecules during extraction. However, for studies using rumen content there is no consensus on (1 the amount of rumen digesta to use or (2 the amount of TRIzol reagent to be used in RNA extraction procedures. This study evaluated the effect of using various quantities of ground rumen digesta and of TRIzol reagent on the yield and quality of extracted RNA. It also investigated the possibility of using lower masses of solid-phase rumen digesta and lower amounts of TRIzol reagent than is used currently, for extraction of RNA for metatranscriptomic studies. We found that high quality RNA could be isolated from 2 g of ground rumen digesta sample, whilst using 0.6 g of ground matter for RNA extraction and using 3 mL (a 5:1 TRIzol : extraction mass ratio of TRIzol reagent. This represents a significant savings in the cost of RNA isolation. These lower masses and volumes were then applied in the RNA-Seq analysis of solid-phase rumen samples obtained from 6 Angus X Hereford beef heifers which had been fed a high forage diet (comprised of barley straw in a forage-to-concentrate ratio of 70:30 for 102 days. A bioinformatics analysis pipeline was developed in-house that generated relative abundance values of archaea, protozoa, fungi and bacteria in the rumen and also allowed the extraction of individual rRNA variable regions that could be analyzed in downstream molecular ecology programs. The average relative abundances of rRNA transcripts of archaea, bacteria

Total rRNA-Seq Analysis Gives Insight into Bacterial, Fungal, Protozoal and Archaeal Communities in the Rumen Using an Optimized RNA Isolation Method.

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Elekwachi, Chijioke O; Wang, Zuo; Wu, Xiaofeng; Rabee, Alaa; Forster, Robert J

2017-01-01

Advances in high throughput, next generation sequencing technologies have allowed an in-depth examination of biological environments and phenomena, and are particularly useful for culture-independent microbial community studies. Recently the use of RNA for metatranscriptomic studies has been used to elucidate the role of active microbes in the environment. Extraction of RNA of appropriate quality is critical in these experiments and TRIzol reagent is often used for maintaining stability of RNA molecules during extraction. However, for studies using rumen content there is no consensus on (1) the amount of rumen digesta to use or (2) the amount of TRIzol reagent to be used in RNA extraction procedures. This study evaluated the effect of using various quantities of ground rumen digesta and of TRIzol reagent on the yield and quality of extracted RNA. It also investigated the possibility of using lower masses of solid-phase rumen digesta and lower amounts of TRIzol reagent than is used currently, for extraction of RNA for metatranscriptomic studies. We found that high quality RNA could be isolated from 2 g of ground rumen digesta sample, whilst using 0.6 g of ground matter for RNA extraction and using 3 mL (a 5:1 TRIzol : extraction mass ratio) of TRIzol reagent. This represents a significant savings in the cost of RNA isolation. These lower masses and volumes were then applied in the RNA-Seq analysis of solid-phase rumen samples obtained from 6 Angus X Hereford beef heifers which had been fed a high forage diet (comprised of barley straw in a forage-to-concentrate ratio of 70:30) for 102 days. A bioinformatics analysis pipeline was developed in-house that generated relative abundance values of archaea, protozoa, fungi and bacteria in the rumen and also allowed the extraction of individual rRNA variable regions that could be analyzed in downstream molecular ecology programs. The average relative abundances of rRNA transcripts of archaea, bacteria, protozoa and fungi in

Influence of probiotics on rumen liquor characteristics and ...

African Journals Online (AJOL)

Probiotics has been noted to work synergistically with rumen microbes and improved rumen liquor characteristics. In this study, we investigated the effect of probiotics inclusion on rumen liquor characteristics (physical, chemical and fermentative qualities) and microbiology in WAD goats. In a completely randomised design, ...

Comparison of rumen bacterial communities in dairy herds of different production.

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Indugu, Nagaraju; Vecchiarelli, Bonnie; Baker, Linda D; Ferguson, James D; Vanamala, Jairam K P; Pitta, Dipti W

2017-08-30

The purpose of this study was to compare the rumen bacterial composition in high and low yielding dairy cows within and between two dairy herds. Eighty five Holstein dairy cows in mid-lactation (79-179 days in milk) were selected from two farms: Farm 12 (M305 = 12,300 kg; n = 47; 24 primiparous cows, 23 multiparous cows) and Farm 9 (M305 = 9700 kg; n = 38; 19 primiparous cows, 19 multiparous cows). Each study cow was sampled once using the stomach tube method and processed for 16S rRNA gene amplicon sequencing using the Ion Torrent (PGM) platform. Differences in bacterial communities between farms were greater (Adonis: R 2  = 0.16; p forage type and proportion in the diets. A combination of corn silage and alfalfa silage may have contributed to the increased proportion of Proteobacteria in Farm 12. It was concluded that Farm 12 had a greater proportion of specialist bacteria that have the potential to enhance rumen fermentative digestion of feedstuffs to support higher milk yields.

Gene expression in bovine rumen epithelium during weaning indentifies molecular regulators of rumen development and growth

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During weaning, rumen epithelial cell function must transition from a pre-ruminant to a true ruminant state for efficient nutrient absorption and metabolism. During this time, the rumen increases from 30 to 70% of the capacity of the gut, significantly impacting net efficiency of feed conversion in ...

Rumen Biohydrogenation and Microbial Community Changes Upon Early Life Supplementation of 22:6n-3 Enriched Microalgae to Goats

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Lore Dewanckele

2018-03-01

Full Text Available Dietary supplementation of docosahexaenoic acid (DHA-enriched products inhibits the final step of biohydrogenation in the adult rumen, resulting in the accumulation of 18:1 isomers, particularly of trans(t-11 18:1. Occasionally, a shift toward the formation of t10 intermediates at the expense of t11 intermediates can be triggered. However, whether similar impact would occur when supplementing DHA-enriched products during pregnancy or early life remains unknown. Therefore, the current in vivo study aimed to investigate the effect of a nutritional intervention with DHA in the early life of goat kids on rumen biohydrogenation and microbial community. Delivery of DHA was achieved by supplementing DHA-enriched microalgae (DHA Gold either to the maternal diet during pregnancy (prenatal or to the diet of the young offspring (postnatal. At the age of 12 weeks, rumen fluid was sampled for analysis of long-chain fatty acids and microbial community based on bacterial 16S rRNA amplicon sequencing. Postnatal supplementation with DHA-enriched microalgae inhibited the final biohydrogenation step, as observed in adult animals. This resulted particularly in increased ruminal proportions of t11 18:1 rather than a shift to t10 intermediates, suggesting that both young and adult goats might be less prone to dietary induced shifts toward the formation of t10 intermediates, in comparison with cows. Although Butyrivibrio species have been identified as the most important biohydrogenating bacteria, this genus was more abundant when complete biohydrogenation, i.e. 18:0 formation, was inhibited. Blautia abundance was positively correlated with 18:0 accumulation, whereas Lactobacillus spp. Dialister spp. and Bifidobacterium spp. were more abundant in situations with greater t10 accumulation. Extensive comparisons made between current results and literature data indicate that current associations between biohydrogenation intermediates and rumen bacteria in young goats

The complete genome sequence of Eubacterium limosum SA11, a metabolically versatile rumen acetogen.

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Kelly, William J; Henderson, Gemma; Pacheco, Diana M; Li, Dong; Reilly, Kerri; Naylor, Graham E; Janssen, Peter H; Attwood, Graeme T; Altermann, Eric; Leahy, Sinead C

2016-01-01

Acetogens are a specialized group of anaerobic bacteria able to produce acetate from CO2 and H2 via the Wood-Ljungdahl pathway. In some gut environments acetogens can compete with methanogens for H2, and as a result rumen acetogens are of interest in the development of microbial approaches for methane mitigation. The acetogen Eubacterium limosum SA11 was isolated from the rumen of a New Zealand sheep and its genome has been sequenced to examine its potential application in methane mitigation strategies, particularly in situations where hydrogenotrophic methanogens are inhibited resulting in increased H2 levels in the rumen. The 4.15 Mb chromosome of SA11 has an average G + C content of 47 %, and encodes 3805 protein-coding genes. There is a single prophage inserted in the chromosome, and several other gene clusters appear to have been acquired by horizontal transfer. These include genes for cell wall glycopolymers, a type VII secretion system, cell surface proteins and chemotaxis. SA11 is able to use a variety of organic substrates in addition to H2/CO2, with acetate and butyrate as the principal fermentation end-products, and genes involved in these metabolic pathways have been identified. An unusual feature is the presence of 39 genes encoding trimethylamine methyltransferase family proteins, more than any other bacterial genome. Overall, SA11 is a metabolically versatile organism, but its ability to grow on such a wide range of substrates suggests it may not be a suitable candidate to take the place of hydrogen-utilizing methanogens in the rumen.

Pasture Feeding Changes the Bovine Rumen and Milk Metabolome

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Tom F. O’Callaghan

2018-04-01

Full Text Available The purpose of this study was to examine the effects of two pasture feeding systems—perennial ryegrass (GRS and perennial ryegrass and white clover (CLV—and an indoor total mixed ration (TMR system on the (a rumen microbiome; (b rumen fluid and milk metabolome; and (c to assess the potential to distinguish milk from different feeding systems by their respective metabolomes. Rumen fluid was collected from nine rumen cannulated cows under the different feeding systems in early, mid and late lactation, and raw milk samples were collected from ten non-cannulated cows in mid-lactation from each of the feeding systems. The microbiota present in rumen liquid and solid portions were analysed using 16S rRNA gene sequencing, while 1H-NMR untargeted metabolomic analysis was performed on rumen fluid and raw milk samples. The rumen microbiota composition was not found to be significantly altered by any feeding system in this study, likely as a result of a shortened adaptation period (two weeks’ exposure time. In contrast, feeding system had a significant effect on both the rumen and milk metabolome. Increased concentrations of volatile fatty acids including acetic acid, an important source of energy for the cow, were detected in the rumen of TMR and CLV-fed cows. Pasture feeding resulted in significantly higher concentrations of isoacids in the rumen. The ruminal fluids of both CLV and GRS-fed cows were found to have increased concentrations of p-cresol, a product of microbiome metabolism. CLV feeding resulted in increased rumen concentrations of formate, a substrate compound for methanogenesis. The TMR feeding resulted in significantly higher rumen choline content, which contributes to animal health and milk production, and succinate, a product of carbohydrate metabolism. Milk and rumen-fluids were shown to have varying levels of dimethyl sulfone in each feeding system, which was found to be an important compound for distinguishing between the diets

Characterization of rumen bacterial strains isolated from enrichments of rumen content in the presence of propolis.

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de Aguiar, Sílvia Cristina; Zeoula, Lucia Maria; do Prado, Odimari Pricila Pires; Arcuri, Pedro Braga; Forano, Evelyne

2014-11-01

Propolis presents many biological properties, including antibacterial activities, and has been proposed as an additive in ruminant nutrition. Twenty bacterial strains, previously isolated from enrichments of Brazilian cow rumen contents in the presence of different propolis extracts (LLOS), were characterized using phenotyping and 16S rRNA identification. Seven strains were assigned to Streptococcus sp., most likely S. bovis, and were all degrading starch. One amylolytic lactate-utilizing strain of Selenomonas ruminantium was also found. Two strains of Clostridium bifermentans were identified and showed proteolytic activity. Two strains were assigned to Mitsuokella jalaludinii and were saccharolytic. One strain belonged to a Bacillus species and seven strains were affiliated with Escherichia coli. All of the 20 strains were able to use many sugars, but none of them were able to degrade the polysaccharides carboxymethylcellulose and xylans. The effect of three propolis extracts (LLOS B1, C1 and C3) was tested on the in vitro growth of four representative isolates of S. bovis, E. coli, M. jalaludinii and C. bifermentans. The growth of S. bovis, E. coli and M. jalaludinii was not affected by the three propolis extracts at 1 mg ml(-1). C. bifermentans growth was completely inhibited at this LLOS concentration, but this bacterium was partially resistant at lower concentrations. LLOS C3, with the lower concentration of phenolic compounds, was a little less inhibitory than B1 and C1 on this strain.

The effects of different levels of sodium caseinate on rumen ...

African Journals Online (AJOL)

CN significantly affected the concentrations of rumen ammonia nitrogen (NH3-N), rumen peptide nitrogen (Pep-N) and the ratio of rumen ammonia nitrogen/ rumen peptide nitrogen (P < 0.05) and consequently blood urea nitrogen, milk urea nitrogen and urinary urea nitrogen concentrations. However digestibility of dry ...

Effect of γ-Aminobutyric Acid (GABA Producing Bacteria on Rumen Fermentation, Biogenic Amine Production and Anti-oxidation Using Corn Meal as Substrate

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Bum Seung Ku

2013-06-01

Full Text Available The effects and significance of γ-amino butyric acid (GABA producing bacteria (GPB on in vitro rumen fermentation and reduction of biogenic amines (histamine, methylamine, ethylamine, and tyramine using corn meal as a substrate were determined. Ruminal samples collected from ruminally fistulated Holstein cows served as inoculum and corn was used as substrate at 2% dry matter (DM. Different inclusion rates of GPB and GABA were evaluated. After incubation, addition of GPB had no significant effect on in vitro fermentation pH and total gas production, but significantly increased the ammonia nitrogen (NH3-N concentration and reduced the total biogenic amines production (p<0.05. Furthermore, antioxidation activity was improved as indicated by the significantly higher concentration of superoxide dismutase (SOD and glutathione peroxidase (GSH-Px among treated samples when compared to the control (p<0.05. Additionally, 0.2% GPB was established as the optimum inclusion level. Taken together, these results suggest the potential of utilizing GPB as feed additives to improve growth performance in ruminants by reducing biogenic amines and increasing anti-oxidation.

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

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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.

Resistance of soil-bound prions to rumen digestion.

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Samuel E Saunders

Full Text Available Before prion uptake and infection can occur in the lower gastrointestinal system, ingested prions are subjected to anaerobic digestion in the rumen of cervids and bovids. The susceptibility of soil-bound prions to rumen digestion has not been evaluated previously. In this study, prions from infectious brain homogenates as well as prions bound to a range of soils and soil minerals were subjected to in vitro rumen digestion, and changes in PrP levels were measured via western blot. Binding to clay appeared to protect noninfectious hamster PrP(c from complete digestion, while both unbound and soil-bound infectious PrP(Sc proved highly resistant to rumen digestion. In addition, no change in intracerebral incubation period was observed following active rumen digestion of unbound hamster HY TME prions and HY TME prions bound to a silty clay loam soil. These results demonstrate that both unbound and soil-bound prions readily survive rumen digestion without a reduction in infectivity, further supporting the potential for soil-mediated transmission of chronic wasting disease (CWD and scrapie in the environment.

Resistance of Soil-Bound Prions to Rumen Digestion

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Saunders, Samuel E.; Bartelt-Hunt, Shannon L.; Bartz, Jason C.

2012-01-01

Before prion uptake and infection can occur in the lower gastrointestinal system, ingested prions are subjected to anaerobic digestion in the rumen of cervids and bovids. The susceptibility of soil-bound prions to rumen digestion has not been evaluated previously. In this study, prions from infectious brain homogenates as well as prions bound to a range of soils and soil minerals were subjected to in vitro rumen digestion, and changes in PrP levels were measured via western blot. Binding to clay appeared to protect noninfectious hamster PrPc from complete digestion, while both unbound and soil-bound infectious PrPSc proved highly resistant to rumen digestion. In addition, no change in intracerebral incubation period was observed following active rumen digestion of unbound hamster HY TME prions and HY TME prions bound to a silty clay loam soil. These results demonstrate that both unbound and soil-bound prions readily survive rumen digestion without a reduction in infectivity, further supporting the potential for soil-mediated transmission of chronic wasting disease (CWD) and scrapie in the environment. PMID:22937149

Ginkgo fruit extract as an additive to modify rumen microbiota and fermentation and to mitigate methane production.

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Oh, S; Shintani, R; Koike, S; Kobayashi, Y

2017-03-01

Ginkgo fruit, an unused byproduct of the ginkgo nut industry, contains antimicrobial compounds known as anacardic acids. Two major cultivars of ginkgo, Kyuju (K) and Tokuro (T), were evaluated for their potential as a feed additive for ruminants. In batch culture, we incubated a mixture of hay and concentrate in diluted rumen fluid with or without 1.6% (fruit equivalent) ginkgo fruit extract. We conducted another series of batch culture studies to determine the dose response of fermentation. We also conducted continuous culture using the rumen simulation technique (RUSITEC) with cultivar K and carried out a pure culture study to monitor the sensitivity of 17 representative rumen bacterial species to ginkgo extract and component phenolics. Although both K and T extracts led to decreased methane and increased propionate production, changes were more apparent with K extract, and were dose-dependent. Total gas production was depressed at doses ≥3.2%, suggesting that 1.6% was the optimal supplementation level. In RUSITEC fermentation supplemented with 1.6% ginkgo K, methane decreased by 53% without affecting total gas or total VFA production, but with decreased acetate and increased propionate. Disappearance of dry matter, neutral detergent fiber, and acid detergent fiber were not affected by ginkgo, but ammonia levels were decreased. Quantitative PCR indicated that the abundance of protozoa, fungi, methanogens, and bacteria related to hydrogen and formate production decreased, but the abundance of bacteria related to propionate production increased. MiSeq analysis (Illumina Inc., San Diego, CA) confirmed these bacterial changes and identified archaeal community changes, including a decrease in Methanobrevibacter and Methanomassiliicoccaceae and an increase in Methanoplanus. Pure culture study results supported the findings for the above bacterial community changes. These results demonstrate that ginkgo fruit can modulate rumen fermentation toward methane mitigation

Studies on the Influence of Tritium Radiation on Anaerobic Bacteria from the Bovine Rumen; Influence de l'Irradiation par le Tritium sur les Bacteries Anaerobies du Rumen des Bovins; 0418 0441 0414 ; Estudios sobre los Efectos de las Radiaciones del Tritio en las Bacterias Anaerobias de la Panza de los Bovinos

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Brueggemann, J.; Giesecke, D. [Institute of Physiology and Animal Nutrition, University of Munich, Munich, Federal Republic of Germany (Germany)

1962-02-15

The bacterial flora in the bovine rumen is mainly composed of strictly anaerobic species supplying the host with large amounts of volatile fatty acids (VFA's) as the main energy source. Long time in-vitro irradiations of the flora with T{sub 2}O in a so-called ''artificial rumen'' under anaerobic conditions have shown that doses up to 75 Krad had no inhibitory effect on bacterial growth and VFA-production. Stimulatory effects resulting in acceleration of cell division and increased amounts of VFA's were observed after irradiating the resting flora at 0 Degree-Sign C with a total dose of about 300 Krad within 20 d, but reproducibility was unsatisfactory because of the heterogeneity of the material. Further experiments were carried out on pure cultures of selected strains of rumen bacteria. Doses up to 150 Krad (5 d) did not markedly influence anaerobic growth and carbohydrate metabolism. However, if oxygen was present during the irradiation of resting cells a strong reduction in viable numbers was noted. Oxygen per se was found to exert a lethal effect on these species following a logarithmic order of decline. It is assumed that secondary radiation products, especially H{sub 2}O{sub 2}, are mainly responsible for the action of T{sub 2}O on these microorganisms which are devoid of catalase. Some confirmatory results are still to be obtained. (author) [French] La flore bacterienne du rumen des bovins se compose surtout d'especes strictement anaerobies qui fournissent a l'hote, comme principale source d'energie, de grandes quantites d'acides gras volatils (AGV). Des irradiations de longue duree de la flore par T{sub 2}O, faites in vitro dans un 'rumen artificiel', en anaerobie, ont montre que des doses allant jusqu'a 75 krads n'ont aucun effet inhibiteur sur la croissance des bacteries et sur la production d'AGV. Des effets stimulateurs, se traduisant par une division cellulaire acceleree et par une production accrue d'AGV, ont ete observes apres irradiation de la

Effect of Tree Leaves on Rumen Fermentation, Microbial Count and Blood Urea Nitrogen of West African Dwarf Goats

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Adelusi, O. O.

2016-12-01

Full Text Available This experiment was carried out to assess the effect of Azadirachta indica, Newbouldia laevis and Spondias mombin leaves on rumen fermentation, microbial count and blood urea nitrogen (BUN of West African Dwarf (WAD goats. Sixteen WAD bucks (11.6 ? 0.9 kg in body weight were allocated to 4 treatments: 1 Control and 2 40 g/day of Azadirachta indica, 3 40 g/day of Newbouldia laevis and 4 40 g/day of Spondias mombin leaves arranged in a completely randomised design. The ground leaves were included in concentrate diets served on dry matter basis at 2% of body weight while Panicum maximum was fed ad libitum. The control diet had no tree leaves. Data were collected on chemical composition, rumen fermentation and microbial ecology, and BUN. Saponin was highest (P < 0.05 in S. mombin (8.14% while A. indica and N. laevis had 5.78% and 1.56%, respectively. Rumen ammonia nitrogen was least (P < 0.05 in goats fed A. indica (8.35 mg/dL while the highest (P < 0.05 total volatile fatty acid (TVFA was obtained from goats fed S. mombin with 125.51 mM. Goats fed N. laevis yielded the highest (P < 0.05 acetate with 70.65 mol/100 mol while propionate production was highest (P < 0.05 in the rumen of goats fed S. mombin (27.15 mol/100 mol. Viable bacteria count was lowest (P < 0.05 in rumen of goats fed A. indica (3.95?1012 cfu/ml while the least (P < 0.05 protozoa population was obtained from the rumen of bucks fed S. mombin (4.18?109 cfu/ml. All goats in the treatments containing tree leaves had higher (P < 0.05 and a rapid increase in BUN between 0 and 6 h post feeding when compared with the Control. It is concluded that feeding ground leaves of S. mombin to goats increases rumen total volatile fatty acid and propionate production and reduces the protozoa population.

Histogenesis of rumen in one-humped camel (Camelus dromedarius

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E. Salimi Naghani* and L. Akradi1

2012-05-01

Full Text Available The aim of this study was to follow several sequence histological changes that occur during the histogenesis of the rumen in one-humped camel (Camelus dromedarius. Histogenesis study was carried out on 66 fetuses of camel from 50th day of gestation until birth (390 days, according to the most relevant histo-differentiation characteristics of the rumen in fetuses, these were divided into four groups: group I (5-24 cm crown-rump length (C-RL; 50-140 days; group II (24-30 cm C-RL; 140-160 days; group III (30-60 cm C-RL; 160-250 days; group IV (60-108 cm C-RL; 250-390 days. At 50 days, the rumen consisted of four layers: the epithelial layer, propria-submucosa, tunica muscularis and serosa. The epithelium glandular region was pseudostratified and in non-glandular region was stratified. The muscularis mucosa was observed incompletely from 140 days between lamina propria and submucosa in glandular region of the rumen to the birth day. The primary lymphatic nodules appeared in lamina propria of glandular region of the rumen at 160 days of gestation. The epithelium of the glandular region in rumen was formed by a simple columnar layer at 250 days. In all groups, the tunica muscularis layer of rumen was increased with ruminal development, gradually. The non-glandular region of rumen was formed by a stratified epithelium and number of these cells increased with ruminal development. The lymphatic nodules and muscularis mucosa in non-glandular region did not observe in all groups. The study observations revealed that non-glandular region of the rumen in the fetuses of camel are less precocious than the rumen of the domestic ruminants.

Microbiota composition, gene pool and its expression in Gir cattle (Bos indicus) rumen under different forage diets using metagenomic and metatranscriptomic approaches.

Science.gov (United States)

Pandit, Ramesh J; Hinsu, Ankit T; Patel, Shriram H; Jakhesara, Subhash J; Koringa, Prakash G; Bruno, Fosso; Psifidi, Androniki; Shah, S V; Joshi, Chaitanya G

2018-03-09

Zebu (Bos indicus) is a domestic cattle species originating from the Indian subcontinent and now widely domesticated on several continents. In this study, we were particularly interested in understanding the functionally active rumen microbiota of an important Zebu breed, the Gir, under different dietary regimes. Metagenomic and metatranscriptomic data were compared at various taxonomic levels to elucidate the differential microbial population and its functional dynamics in Gir cattle rumen under different roughage dietary regimes. Different proportions of roughage rather than the type of roughage (dry or green) modulated microbiome composition and the expression of its gene pool. Fibre degrading bacteria (i.e. Clostridium, Ruminococcus, Eubacterium, Butyrivibrio, Bacillus and Roseburia) were higher in the solid fraction of rumen (Pcomparison of metagenomic shotgun and metatranscriptomic sequencing appeared to be a much richer source of information compared to conventional metagenomic analysis. Copyright © 2018 Elsevier GmbH. All rights reserved.

Nitrogen metabolism in the rumen and its measurement

International Nuclear Information System (INIS)

Nolan, J.V.; Leng, R.A.

1983-01-01

Methods are needed to maximize the synthesis of microbial protein in the rumen from readily available, inexpensive (usually non-protein N) sources and thereby to reduce the requirements for true protein in the diet. Some currently available in vitro and in vivo methods for estimating microbial protein synthesis in the rumen are discussed. The factors that alter maintenance ATP requirements of microorganisms, and thereby potentially alter the efficiency of cell growth per unit of fermented organic matter (Ysub(ATP)), are discussed: e.g. continuity and level of supply of substrates; dietary and recycled N and other nutrients; dilution rate; the presence of large protozoal populations in the rumen; cell lysis and N cycling in the rumen. Quantitative studies of these factors have been made by using a variety of isotope tracer techniques and have been applied to a quantitative model of N transactions in the rumen. (author)

Dynamics of initial colonization of nonconserved perennial ryegrass by anaerobic fungi in the bovine rumen.

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Edwards, Joan E; Kingston-Smith, Alison H; Jimenez, Hugo R; Huws, Sharon A; Skøt, Kirsten P; Griffith, Gareth W; McEwan, Neil R; Theodorou, Michael K

2008-12-01

Anaerobic fungi (Neocallimastigales) are active degraders of fibrous plant material in the rumen. However, only limited information is available relating to how quickly they colonize ingested feed particles. The aim of this study was to determine the dynamics of initial colonization of forage by anaerobic fungi in the rumen and the impact of different postsampling wash procedures used to remove loosely associated microorganisms. Neocallimastigales-specific molecular techniques were optimized to ensure maximal coverage before application to assess the population size (quantitative PCR) and composition (automated ribosomal intergenic spacer analysis) of the colonizing anaerobic fungi. Colonization of perennial ryegrass (PRG) was evident within 5 min, with no consistent effect of time or wash procedure on fungal population composition. Wash procedure had no effect on population size unlike time, which had a significant effect. Colonizing fungal population size continued to increase over the incubation period after an initial lag of c. 4 min. This dynamic differs from that reported previously for rumen bacteria, where substantial colonization of PRG occurred within 5 min. The observed delay in colonization of plant material by anaerobic fungi is suggested to be primarily mediated by the time taken for fungal zoospores to locate, attach and encyst on plant material.

TWO-COMPONENT SYSTEM: A MOLECULAR DIALOGUE BETWEEN RUMINAL BACTERIA AND FEED PARTICLES (FORAGE PLANTS

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Monica Marcela Galicia Jimenez

2017-12-01

Full Text Available The ability to adapt rapidly to changes in the environment is one of the main characteristics of the bacterial cell. The rumen is a highly dynamic environment, and none of the changes are permanent due to the various microbial species found in the rumen. Signal transduction networks are information processing pathways that recognize various physical and chemical stimuli, amplification, signal processing, and trigger responses of the bacterial cell. The aim of the present review is to show the importance of these two component systems in rumen bacteria, because it is based on the knowledge of the principles governing the bacterial population communication, its main interactions and products of metabolism, we can approach the manipulation of Ruminal fermentation to improve animal health, productivity and food safety.

Rumen distension and contraction influence feed preference by sheep.

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Villalba, J J; Provenza, F D; Stott, R

2009-01-01

Distension of the rumen limits feed intake by livestock. Ruminal dysfunctions due to bloat, which causes distension by accumulation of excessive gas within the rumen, also reduce feeding. We hypothesized that excessive levels of rumen distension cause feed aversions and that preference increases for feeds eaten in association with recovery from bloat. To test these hypotheses, we determined whether 12 commercial crossbred lambs (average initial BW of 43 +/- 2 kg) could associate ingestion of specific feeds with the consequences of increased intraruminal pressure and its subsidence. Six of the lambs were fitted with rumen cannulas and offered ground alfalfa for 30 min after a rubber balloon was inserted into the rumen of each animal and distended with air to volumes of 1.8, 2.5, or 4.5 L. Subsequently, balloons were deflated and alfalfa was offered again for a second period of 30 min. Feed intake was not affected when the balloon was not distended (P = 0.45 to 0.93), but distension reduced feed intake (P rumen distension (P = 0.17 to P = 0.87). Thus, rumen distension and recovery from distension induced feed aversions and preferences, respectively, which may be critical in learning avoidance of bloat-inducing plants and preferences for plants and supplements that relieve the incidence of bloat.

The contribution of mathematical modeling to understanding the dynamic aspects of rumen metabolism

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André Bannink

2016-11-01

Full Text Available All rumen models cover the main drivers of variation in rumen function, which are feed intake, the differences between feedstuffs and feeds in their intrinsic rumen degradation characteristics, and fractional outflow rate of fluid and particulate matter. Dynamic modeling approaches are best suited to the prediction of more nuanced responses in rumen metabolism, and represent the dynamics of the interaction between substrates and micro-organisms and inter-microbial interactions. The concepts of dynamics are discussed for the case of rumen starch digestion as influenced by starch intake rate and frequency of feed intake, and for the case of fermentation of fiber in the large intestine. Adding representations of new functional classes of micro-organisms (i.e. with new characteristics from the perspective of whole rumen function in rumen models only delivers new insights if complemented by the dynamics of their interactions with other functional classes. Rumen fermentation conditions have to be represented due to their profound impact on the dynamics of substrate degradation and microbial metabolism. Although the importance of rumen acidity is generally acknowledged, more emphasis is needed on predicting its variation as well as variation in the processes that underlie rumen fluid dynamics. The rumen wall has an important role in adapting to rapid changes in the rumen environment, clearing of volatile fatty acids (VFA, and maintaining rumen pH within limits. Dynamics of rumen wall epithelia and its role in VFA absorption needs to be better represented in models which aim to predict rumen responses across nutritional or physiological states. For a detailed prediction of rumen N balance there is merit in a dynamic modeling approach compared to the static approaches adopted in current protein evaluation systems. Improvement is needed on previous attempts to predict rumen VFA profiles, and this should be pursued by introducing factors that relate more

Microbiological and biochemical response of certain proteolytic bacterial isolates to varying levels of gamma irradiation

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El-Hifnawi, H.M.N.E.A.

1997-01-01

Amniotic membrane allo - and xeno grafts prepared from human foetal placenta, and their potential replacement of skin autotransplant, would significantly contribute to the success of clinical treatment of skin burns. Allo-and xenografts of human amniotic membrane should be ensured for their sterility, bio-mechanics and tissue antigenicity. The present study has been focused on sterilization and sterility assurance of the membrane grafts. Physico-chemical properties and antigenicity of the grafts await investigation. In the present study the isolation and identification of the bacteria contaminating the amniotic membrane allo-and xenografts prepared from human foetal placenta and the effect of gamma irradiation on it has been investigated. The proteolytic activity of these bacteria and the role of gamma irradiation in the control of bacterial activity were similarly investigated

Methane Production in Dairy Cows Correlates with Rumen Methanogenic and Bacterial Community Structure.

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Danielsson, Rebecca; Dicksved, Johan; Sun, Li; Gonda, Horacio; Müller, Bettina; Schnürer, Anna; Bertilsson, Jan

2017-01-01

Methane (CH 4 ) is produced as an end product from feed fermentation in the rumen. Yield of CH 4 varies between individuals despite identical feeding conditions. To get a better understanding of factors behind the individual variation, 73 dairy cows given the same feed but differing in CH 4 emissions were investigated with focus on fiber digestion, fermentation end products and bacterial and archaeal composition. In total 21 cows (12 Holstein, 9 Swedish Red) identified as persistent low, medium or high CH 4 emitters over a 3 month period were furthermore chosen for analysis of microbial community structure in rumen fluid. This was assessed by sequencing the V4 region of 16S rRNA gene and by quantitative qPCR of targeted Methanobrevibacter groups. The results showed a positive correlation between low CH 4 emitters and higher abundance of Methanobrevibacter ruminantium clade. Principal coordinate analysis (PCoA) on operational taxonomic unit (OTU) level of bacteria showed two distinct clusters ( P microbial population or host genetic differences that is reflected in bacterial and archaeal (or methanogens) populations.

Effect of Dietary Chestnut or Quebracho Tannin Supplementation on Microbial Community and Fatty Acid Profile in the Rumen of Dairy Ewes

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Arianna Buccioni

2017-01-01

Full Text Available Ruminants derived products have a prominent role in diets and economy worldwide; therefore, the capability to control the rumen microbial ecosystem, for ameliorating their quality, is of fundamental importance in the livestock sector. The aim of this study was to evaluate the effect of dietary supplementation with chestnut and quebracho tannins on microbial community and fatty acid profile, in the rumen fluid of dairy ewes. Multivariate analysis of PCR-DGGE profiles of rumen microbial communities showed a correlation among the presence of chestnut or quebracho in the diet, the specific Butyrivibrio group DGGE profiles, the increase in 18:3 cis9, cis12, and cis15; 18:2 cis9 and cis12; 18:2 cis9 and trans11; 18:2 trans11 and cis15; and 18:1 trans11 content, and the decrease in 18:0 concentration. Phylogenetic analysis of DGGE band sequences revealed the presence of bacteria representatives related to the genera Hungatella, Ruminococcus, and Eubacterium and unclassified Lachnospiraceae family members, suggesting that these taxa could be affected by tannins presence in the diets. The results of this study showed that tannins from chestnut and quebracho can reduce the biohydrogenation of unsaturated fatty acids through changes in rumen microbial communities.

Effect of Gynosaponin on Rumen Methanogenesis under Different Forage-Concentrate Ratios

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Bakhetgul Manatbay

2014-08-01

Full Text Available The study aimed to investigate the effects of gynosaponin on in vitro methanogenesis under different forage-concentrate ratios (F:C ratios. Experiment was conducted with two kinds of F:C ratios (F:C = 7:3 and F:C = 3:7 and gynosaponin addition (0 mg and 16 mg in a 2×2 double factorial design. In the presence of gynosaponin, methane production and acetate concentration were significantly decreased, whereas concentration of propionate tended to be increased resulting in a significant reduction (p<0.05 of acetate:propionate ratio (A:P ratio, in high-forage substrate. Gynosaponin treatment increased (p<0.05 the butyrate concentration in both F:C ratios. Denaturing gradient gel electrophoresis (DGGE analysis showed there was no apparent shift in the composition of total bacteria, protozoa and methanogens after treated by gynosaponin under both F:C ratios. The real-time polymerase chain reaction (PCR analysis indicated that variable F:C ratios significantly affected the abundances of Fibrobacter succinogenes, Rumninococcus flavefaciens, total fungi and counts of protozoa (p<0.05, but did not affect the mcrA gene copies of methanogens and abundance of total bacteria. Counts of protozoa and abundance of F.succinogenes were decreased significantly (p<0.05, whereas mcrA gene copies of methanogens were decreased slightly (p<0.10 in high-forage substrate after treated by gynosaponin. However, gynosaponin treatment under high-concentrate level did not affect the methanogenesis, fermentation characteristics and tested microbes. Accordingly, overall results suggested that gynosaponin supplementation reduced the in vitro methanogenesis and improved rumen fermentation under high-forage condition by changing the abundances of related rumen microbes.

Research Applications of Proteolytic Enzymes in Molecular Biology

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József Tőzsér

2013-11-01

Full Text Available Proteolytic enzymes (also termed peptidases, proteases and proteinases are capable of hydrolyzing peptide bonds in proteins. They can be found in all living organisms, from viruses to animals and humans. Proteolytic enzymes have great medical and pharmaceutical importance due to their key role in biological processes and in the life-cycle of many pathogens. Proteases are extensively applied enzymes in several sectors of industry and biotechnology, furthermore, numerous research applications require their use, including production of Klenow fragments, peptide synthesis, digestion of unwanted proteins during nucleic acid purification, cell culturing and tissue dissociation, preparation of recombinant antibody fragments for research, diagnostics and therapy, exploration of the structure-function relationships by structural studies, removal of affinity tags from fusion proteins in recombinant protein techniques, peptide sequencing and proteolytic digestion of proteins in proteomics. The aim of this paper is to review the molecular biological aspects of proteolytic enzymes and summarize their applications in the life sciences.

RUMINANT NUTRITION SYMPOSIUM: How to use data on the rumen microbiome to improve our understanding of ruminant nutrition.

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Firkins, J L; Yu, Z

2015-04-01

Metagenomics and high-throughput sequencing have greatly expanded our knowledge of the rumen microbiome. Surveys of all 4 cellular microbial groups (bacteria, archaea, protozoa, and fungi) reveal profound diversity. Even so, evidence exists for core members to perform key degradative or fermentative roles for the host. Some core members are functionally similar yet taxonomically diverse, and noncore members are particularly diverse and probably vary among diets, animals, and over time after feeding. Gains in functional knowledge are being made and offer much potential not only to improve fiber digestibility, decrease methane emissions, and improve efficiency of nitrogen usage but also to help explain the differences in nutrient digestibility or feed efficiency among animals fed the same diet. Integrated research using metagenomics, bioinformatics, traditional ruminant nutrition, and statistical inferences have provided opportunities for ruminant nutritionists and rumen microbiologists to work synergistically to improve nutrient utilization efficiency while minimizing output of wastes and emissions of methane and ammonia. Examples we highlight include residual feed intake, rumen biohydrogenation of unsaturated fatty acids, and dietary inclusion of ionophores. However, there are still some quantitative limitations in approaches being used. This review addresses knowledge gained and current limitations and challenges that remain.

Phylogenetic analysis of methanogens from the bovine rumen

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Forster Robert J

2001-05-01

Full Text Available Abstract Background Interest in methanogens from ruminants has resulted from the role of methane in global warming and from the fact that cattle typically lose 6 % of ingested energy as methane. Several species of methanogens have been isolated from ruminants. However they are difficult to culture, few have been consistently found in high numbers, and it is likely that major species of rumen methanogens are yet to be identified. Results Total DNA from clarified bovine rumen fluid was amplified using primers specific for Archaeal 16S rRNA gene sequences (rDNA. Phylogenetic analysis of 41 rDNA sequences identified three clusters of methanogens. The largest cluster contained two distinct subclusters with rDNA sequences similar to Methanobrevibacter ruminantium 16S rDNA. A second cluster contained sequences related to 16S rDNA from Methanosphaera stadtmanae, an organism not previously described in the rumen. The third cluster contained rDNA sequences that may form a novel group of rumen methanogens. Conclusions The current set of 16S rRNA hybridization probes targeting methanogenic Archaea does not cover the phylogenetic diversity present in the rumen and possibly other gastro-intestinal tract environments. New probes and quantitative PCR assays are needed to determine the distribution of the newly identified methanogen clusters in rumen microbial communities.

A study of rumen water volume, rate of flow of water and rumen dry matter turnover time measurement by using /sup 51/Cr-labelled EDTA

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Krishna, G; Ekern, A [Agricultural University of Norway. Dept. of Animal Nutrition

1974-06-01

Two fistulated adult sheep were infused with 100 ..mu..Vi /sup 51/Cr-EDTA, four hours after morning feeding, so as to calculate rumen water volume, and rate of flow of water from reticulo-rumen. The average figure of rumen water volume obtained was 2.191 litre, rate of flow of water expressed as volume per cent per hour was 7.55. The biological half-life of marker /sup 51/Cr-EDTA in rumen was 9.34 hours. The percent recovery of infused dosage of /sup 51/Cr-EDTA through feces and urine was 66 and 5 during the period of four days after infusion. Dry matter turnover time in the rumen was 0.483 days.

Effect of Entodinium caudatum on starch intake and glycogen formation by Eudiplodinium maggii in the rumen and reticulum.

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Bełżecki, Grzegorz; McEwan, Neil R; Kowalik, Barbara; Michałowski, Tadeusz; Miltko, Renata

2017-02-01

This study aimed to quantify the engulfed starch and reserve α-glucans (glycogen) in the cells of the ciliates Eudiplodinium maggii, as well the α-glucans in defaunated and selectively faunated sheep. The content of starch inside the cell of ciliates varied from 21 to 183mg/g protozoal DM relative to the rumen fauna composition whereas, the glycogen fluctuated between 17 and 126mg/g dry matter (DM) of this ciliate species. Establishment of the population Entodinium caudatum in the rumen of sheep already faunated with E. maggii caused a drop in both types of quantified carbohydrates. The content of α-glucans in the rumen of defaunated sheep varied from 4.4 to 19.9mg/g DM and increased to 7.4-29.9 or 11.8-33.9mg/g DM of rumen contents in the presence of only E. maggii or E. maggii and E. caudatum, respectively. The lowest content of the carbohydrates was always found just before feeding and the highest at 4h thereafter. The α-glucans in the reticulum varied 7.5-40.1, 14.3-76.8 or 21.9-106.1mg/g DM of reticulum content for defaunated, monofaunated or bifaunated sheep, respectively. The results indicated that both ciliate species engulf starch granules and convert the digestion products to the glycogen, diminishing the pool of starch available for amylolytic bacteria. Copyright © 2016 Elsevier GmbH. All rights reserved.

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

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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, ...

Soybean oil and linseed oil supplementation affect profiles of ruminal microorganisms in dairy cows.

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Yang, S L; Bu, D P; Wang, J Q; Hu, Z Y; Li, D; Wei, H Y; Zhou, L Y; Loor, J J

2009-11-01

The objective of this study was to evaluate changes in ruminal microorganisms and fermentation parameters due to dietary supplementation of soybean and linseed oil alone or in combination. Four dietary treatments were tested in a Latin square designed experiment using four primiparous rumen-cannulated dairy cows. Treatments were control (C, 60 : 40 forage to concentrate) or C with 4% soybean oil (S), 4% linseed oil (L) or 2% soybean oil plus 2% linseed oil (SL) in a 4 × 4 Latin square with four periods of 21 days. Forage and concentrate mixtures were fed at 0800 and 2000 h daily. Ruminal fluid was collected every 2 h over a 12-h period on day 19 of each experimental period and pH was measured immediately. Samples were prepared for analyses of concentrations of volatile fatty acids (VFA) by GLC and ammonia. Counts of total and individual bacterial groups (cellulolytic, proteolytic, amylolytic bacteria and total viable bacteria) were performed using the roll-tube technique, and protozoa counts were measured via microscopy in ruminal fluid collected at 0, 4 and 8 h after the morning feeding. Content of ruminal digesta was obtained via the rumen cannula before the morning feeding and used immediately for DNA extraction and quantity of specific bacterial species was obtained using real- time PCR. Ruminal pH did not differ but total VFA (110 v. 105 mmol/l) were lower (P ruminal NH3-N (4.4 v. 5.6 mmol/l) was greater (P ruminal fluid was substantially lower (P ruminal microorganisms, except proteolytic bacteria, are highly susceptible to dietary unsaturated fatty acids supplementation, particularly when linolenic acid rich oils were fed. Dietary oil effects on ruminal fermentation parameters seemed associated with the profile of ruminal microorganisms.

Rumen bypass nutrients: Manipulation and implications

International Nuclear Information System (INIS)

Leng, R.A.; Nolan, J.V.; Preston, T.R.

1983-01-01

The feeds available for ruminants in developing countries are either agro-industrial by-products or specially grown forage crops. Many of these feeds are low in protein and require supplementation with non-protein N (NPN) to maintain efficient rumen function and digestibility. The principles for utilizing high energy, low protein feeds by ruminants are discussed in relation to the supply of NPN, the establishment of efficient rumen function, maximizing feed intake by means of supplements, and increasing total energy and protein intake by using supplements which bypass the rumen. To illustrate it the application of these principles to feeding systems based on molasses, chopped whole sugar cane and derinded sugar cane is discussed. The implications of the principles in increasing the feeding value of straw are also discussed. (author)

Effects of monolaurin on ruminal methanogens and selected bacterial species from cattle, as determined with the rumen simulation technique.

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Klevenhusen, Fenja; Meile, Leo; Kreuzer, Michael; Soliva, Carla R

2011-10-01

Before being able to implement effective ruminal methane mitigation strategies via feed supplementation, the assessment of side effects on ruminal fermentation and rumen microbial populations is indispensable. In this respect we investigated the effects of monolaurin, a methane-mitigating lipid, on methanogens and important carbohydrate-degrading bacteria present in ruminal fluid of dairy cattle in continuous culture employing the rumen simulation technique. In six experimental runs, each lasting for 10 days, four diets with different carbohydrate composition, based on hay, maize, wheat and a maize-wheat mixture, either remained non-supplemented or were supplemented with monolaurin and incubated in a ruminal-fluid buffer mixture. Incubation liquid samples from days 6 to 10 of incubation were analyzed with relative quantitative polymerase chain reaction (qPCR) of 16S rRNA genes to assess monolaurin-induced shifts in specific rumen microbial populations in relation to the corresponding non-supplemented diets. Monolaurin completely inhibited Fibrobacter succinogenes in all diets while the response of the other cellulolytic bacteria varied in dependence of the diet. Megasphaera elsdenii remained unaffected by monolaurin in the two diets containing maize, but was slightly stimulated by monolaurin with the wheat and largely with the hay diet. The supply of monolaurin suppressed Methanomicrobiales below the detection limit with all diets, whereas relative 16S rRNA gene copy numbers of Methanobacteriales increased by 7-fold with monolaurin in case of the hay diet. Total Archaea were decreased by up to over 90%, but this was significant only for the wheat containing diets. Thus, monolaurin exerted variable effects mediated by unknown mechanisms on important ruminal microbes involved in carbohydrate degradation, along with its suppression of methane formation. The applicability of monolaurin for methane mitigation in ruminants thus depends on the extent to which adverse

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

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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

The Concentrations of Rumen Fluid Volatile Fatty Acids and Ammonia, and Rumen Microbial Protein Production in Sheep Given Feed During the Day and Night Time

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Gumilar, D. A. K. W.; Rianto, E.; Arifin, M.

2018-02-01

An experimental study was carried out to investigate the concentrations of volatile (VFA), ammonia and microbial protein production of rumen fluid in sheep given fedd during the day and at night. This study used 12 fat-tailed rams aged 12-18 months and weighed 24,12 ± 25 kg (CV = 10,51%). The rams were fed a complete feed containing 16.64% protein and 68,33% total digestible nutrients (TDN). The rams were allocated into a completely randomised design with 3 treatments and 4 replications. The treatments applied were: T1: day time feeding (6.00 hrs - 18.00 hrs); T2: night time feeding (18.00 hrs - 6.00 hrs); and T3: day and night time feedings (6.00 hrs - 6.00 hrs). The parameters observed were dry matter intake (DMI), rumen VFA concentration, rumen ammonia concentration, rumen rmicrobial protein production and the efficiency of rumen microbial protein production. The results showed that feeding time did not significantly affect (P>0.05) all the parameters observed. Dry matter intake, VFA concentration, ammonia concentration, the microbial protein production of rumen fluid and the efficiency of microbial protein production were 1,073g/d, 49.69 mmol; 4.77 mg N/100 ml, 12,111 g/d and 19.96 g per kg digestible organic matter intake (DOMI), respectively. It is concluded that feeding time did not affect DMI, condition of rumen fluid and rumen microbial protein production in sheep.

Rumen microbial variation and nutrient utilisation in mithun (Bos frontalis) under different feeding regimes.

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Prakash, B; Saha, S K; Khate, K; Agarwal, N; Katole, S; Haque, N; Rajkhowa, C

2013-04-01

The aim of the study was to investigate the effect of feeding different diets on fermentation, enzyme activities and microbial population in the rumen fluid of mithun (Bos frontalis). In a randomized block design, 20 male mithun (6-8 months of age, 152 ± 12.6 kg body weight) were randomly divided into four experimental groups (n = 5/group) and fed experimental diets ad libitum for 180 days. The diet R1 contained tree foliages (TF), R2 comprised of 50% concentrate mixture (CM) and 50% TF, R3 contained 50% CM and 50% rice straw, and R4 contained 50% CM, 25% TF and 25% rice straw. Rumen liquor was collected at 0 and 180 days of the experiment for estimation of different ruminal parameters and a digestion trial was conducted at the end of the experiment. Rumen fluid was analysed for pH, ammonia nitrogen (NH3 -N), total-N, ruminal enzymes, short chain fatty acid (SCFA) and microbial profile. The relative quantification of ruminal microbes was carried out with real-time PCR using bacteria as the house keeping gene. The dry matter intake, nutrients digestibility, body weight gain, NH3 -N, total-N, carboxymethyl cellulase, avicelase, xylanase, amylase, protease and molar proportion of butyrate were (p ecology, nutrient utilization and thus better performance under stall fed system. © 2012 Blackwell Verlag GmbH.

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.

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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.

SEVILLA and U.M. LUSTRIA. 2006. Changes in rumen ecosystem and feed dry matter degradability of buffalo which received rumen content of cattle through cross inoculation

OpenAIRE

Dicky Pamungkas; Cesar C Sevilla; Ulysses M Lustria

2006-01-01

The research was done to identify changes in rumen ecosystem of buffalo which received rumen content of cattle. As much as three head of fistulated male buffaloes (live weight of 450-550 kg) and three fistulated female cattle (live weight 250-380 kg) were used. This experiment was done three stage as follows: pre-inoculation, inoculation and post-inoculation. In Pre-inoculation, the sample of rumen content was taken two hours before morning feeding and directly observed for pH rumen liquor, a...

Rumen microbial communities influence metabolic phenotypes in lambs

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Diego P. Morgavi

2015-10-01

Full Text Available The rumen microbiota is an essential part of ruminants forging 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.

Modeling the distribution of ciliate protozoa in the reticulo-rumen using linear programming

NARCIS (Netherlands)

Hook, S.E.; Dijkstra, J.; Wright, A.G.; McBride, B.W.; France, J.

2012-01-01

The flow of ciliate protozoa from the reticulo-rumen is significantly less than expected given the total density of rumen protozoa present. To maintain their numbers in the reticulo-rumen, protozoa can be selectively retained through association with feed particles and the rumen wall. Few

Pseudo-affinity chromatography of rumen microbial cellulase on ...

African Journals Online (AJOL)

Pseudo-affinity chromatography of rumen microbial cellulase on Sepharose- Cibacron Blue F3GA. ... African Journal of Biotechnology ... Pseudo affinity adsorption of bioproducts on Sepharose-cibacron blue F3-GA was subjected to rumen microbial enzyme evaluation through batch binding and column chromatography of ...

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

Science.gov (United States)

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

2013-09-01

prefeeding. Seven of the target taxa were more abundant in either the liquid or solid fractions of ruminal digesta. By accounting for the total mass of liquid and solid fractions in the rumen and the relative abundance of total bacteria in each fraction, it was estimated that 92% of total bacteria were found in the solid digesta fraction. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Evaluation Nutritients Of Rice Bran Second Quality Fermented Using Rumen Fluid

OpenAIRE

ermalia, ayu afria ulita

2016-01-01

Rice bran is agriculture  waste that easy to find. Means to increase biological value of rice bran can do with decrease of highly crude fiber. Treatment that to do with fermentation use rumen fluid from cows. Rumen fluid potential is easy to find in slaughterhouse, this rumen fluid much to never utilization. This purpose of the research for evaluation of rice bran nutrition value that fermentating used rumen fluid, with different levels and long time incubations for get lower crude fiber and ...

Characterization of proteolytic and anti-proteolytic activity involvement in sterlet spermatozoon maturation

Czech Academy of Sciences Publication Activity Database

Dzyuba, V.; Słowińska, M.; Cosson, J.; Ciereszko, A.; Boryshpolets, S.; Štěrba, Ján; Rodina, M.; Linhart, O.; Dzyuba, B.

2016-01-01

Roč. 42, č. 6 (2016), s. 1755-1766 ISSN 0920-1742 R&D Projects: GA MŠk(CZ) EE2.3.30.0032 Institutional support: RVO:60077344 Keywords : amidase activity * anti-proteolytic activity * casein and gelatin zymography * siberian sturgeon sperm * spermatozoon maturation * sterlet sperm Subject RIV: ED - Physiology Impact factor: 1.647, year: 2016

Modeling the distribution of ciliate protozoa in the reticulo-rumen using linear programming.

Science.gov (United States)

Hook, S E; Dijkstra, J; Wright, A-D G; McBride, B W; France, J

2012-01-01

The flow of ciliate protozoa from the reticulo-rumen is significantly less than expected given the total density of rumen protozoa present. To maintain their numbers in the reticulo-rumen, protozoa can be selectively retained through association with feed particles and the rumen wall. Few mathematical models have been designed to model rumen protozoa in both the free-living and attached phases, and the data used in the models were acquired using classical techniques. It has therefore become necessary to provide an updated model that more accurately represents these microorganisms and incorporates the recent literature on distribution, sequestration, and generation times. This paper represents a novel approach to synthesizing experimental data on rumen microorganisms in a quantitative and structured manner. The development of a linear programming model of rumen protozoa in an approximate steady state will be described and applied to data from healthy ruminants consuming commonly fed diets. In the model, protozoa associated with the liquid phase and protozoa attached to particulate matter or sequestered against the rumen wall are distinguished. Growth, passage, death, and transfer of protozoa between both pools are represented. The results from the model application using the contrasting diets of increased forage content versus increased starch content indicate that the majority of rumen protozoa, 63 to 90%, are found in the attached phase, either attached to feed particles or sequestered on the rumen wall. A slightly greater proportion of protozoa are found in the attached phase in animals fed a hay diet compared with a starch diet. This suggests that experimental protocols that only sample protozoa from the rumen fluid could be significantly underestimating the size of the protozoal population of the rumen. Further data are required on the distribution of ciliate protozoa in the rumen of healthy animals to improve model development, but the model described herein

Rumen metabolism and recent developments

International Nuclear Information System (INIS)

Smith, R.H.; Oldham, J.D.

1983-01-01

The rumen contains a heterogenous microbial population unevenly distributed in the fluid, in solid fractions of the contents and on the rumen wall. Understanding the ways in which these populations interact with dietary nutrients or those of endogenous origin is essential for improving prediction of nutritional responses, for developing economic production systems and for making maximum use of low quality feeds. Important factors affecting fibre digestion, protein degradation and microbial protein synthesis include turnover rates of individual digesta fractions, long lasting or transient deficiencies or excesses of specific nutrients, and the susceptibilities to microbial attack of naturally occurring or treated dietary components. (author)

The Effect of DNA Extraction Methods on Observed Microbial Communities from Fibrous and Liquid Rumen Fractions of Dairy Cows.

Science.gov (United States)

Vaidya, Jueeli D; van den Bogert, Bartholomeus; Edwards, Joan E; Boekhorst, Jos; van Gastelen, Sanne; Saccenti, Edoardo; Plugge, Caroline M; Smidt, Hauke

2018-01-01

DNA based methods have been widely used to study the complexity of the rumen microbiota, and it is well known that the method of DNA extraction is a critical step in enabling accurate assessment of this complexity. Rumen fluid (RF) and fibrous content (FC) fractions differ substantially in terms of their physical nature and associated microorganisms. The aim of this study was therefore to assess the effect of four DNA extraction methods (RBB, PBB, FDSS, PQIAmini) differing in cell lysis and/or DNA recovery methods on the observed microbial diversity in RF and FC fractions using samples from four rumen cannulated dairy cows fed 100% grass silage (GS100), 67% GS and 33% maize silage (GS67MS33), 33% GS and 67% MS (GS33MS67), or 100% MS (MS100). An ANOVA statistical test was applied on DNA quality and yield measurements, and it was found that the DNA yield was significantly affected by extraction method ( p anaerobic fungal communities using quantitative PCR and pyrosequencing of relevant taxonomic markers. Redundancy analysis (RDA) of bacterial 16S rRNA gene sequence data at the family level showed that there was a significant effect of rumen fraction ( p = 0.012), and that PBB ( p = 0.012) and FDSS ( p = 0.024) also significantly contributed to explaining the observed variation in bacterial community composition. Whilst the DNA extraction method affected the apparent bacterial community composition, no single extraction method could be concluded to be ineffective. No obvious effect of DNA extraction method on the anaerobic fungi or archaea was observed, although fraction effects were evident for both. In summary, the comprehensive assessment of observed communities of bacteria, archaea and anaerobic fungi described here provides insight into a rational basis for selecting an optimal methodology to obtain a representative picture of the rumen microbiota.

A study of rumen water volume, rate of flow of water and rumen dry matter turnover time measurement by using 51Cr-labelled EDTA

International Nuclear Information System (INIS)

Krishna, G.; Ekern, A.

1974-01-01

Two fistulated adult sheep were infused with 100 μVi 51 Cr-EDTA, four hours after morning feeding, so as to calculate fumen water volume, and rate of flow of water from reticulo-rumen. The average figure of rumen water volume obtained was 2.191 litre, rate of flow of water expressed as volume per cent per hour was 7.55. The biological half-life of marker 51 Cr-EDTA in rumen was 9.34 hours. The percent recovery of infused dosage of 51 Cr-EDTA through faeces and urine was 66 and 5 during the period of four days after infusion. Dry matter turnover time in the rumen was 0.483 days. (author)

PENGARUH SUHU DAN KONSENTRASI RUMEN SAPI TERHADAP PRODUKSI BIOGAS DARI VINASSE

Directory of Open Access Journals (Sweden)

Rr. Dewi Artanti Putri

2015-07-01

Full Text Available Vinasse merupakan limbah yang dihasilkan oleh produksi bioetanol yang mempunyai kandungan COD (Chemical Oxygen Demand yang tinggi. Dengan karakteristik tersebut vinasse lebih tepat diuraikan dengan proses anaerob menjadi biogas. Penelitian ini dilakukan untuk mengkaji pengaruh suhu dan perbandingan rumen sapi  yang dibutuhkan untuk mendapatkan biogas dengan hasil yang optimum. Suhu mempengaruhi pertumbuhan mikroorganisme dan kecepatan reaksi dalam pembentukan biogas. Rumen sapi adalah inokulum atau starter yang merupakan bahan yang perlu ditambahkan ke dalam sistem digester biogas. Percobaan dilakukan dalam digester volum 500 ml, dioperasikan pada pH 7 dengan memvariasikan perbandingan suhu,yaitu suhu ruang, suhu 50 oC, dan suhu60 oC dan variasi konsentrasi rumen sapi  5%, 10%, 15%. Proses fermentasi dilakukan dengan cara batch dengan pengukuran gas setiap 2-3 hari menggunakan metode water displacement technique sampai gas tidak terbentuk selama 60 hari. Respon yang diambil pada penelitian ini adalah volume gas yang dihasilkan berdasarkan pengaruh suhu dan konsentrasi rumen sapi terhadap produksi biogas. Perubahan suhu dan konsentrasi rumen sapi sangat mempengaruhi produksi biogas. Hasil yang terbaik dari penelitian ini adalah pada konsentrasi rumen 15% pada suhu ruang yaitu sebanyak 370 ml. Kata kunci: biogas, vinasse, suhu, rumen sapiVinasse is the waste generated by the production of bioethanol which has high content of COD (Chemical Oxygen Demand. With these characteristics, it is more appropriate to convert it into biogas through anaerobic digestion process. This study was conducted to assess the effect of temperature and the cow rumen concentration needed to obtain biogas with optimum results. Temperature affects the growth of microorganisms and speed of reaction in the formation of biogas. The cow rumen was used as inoculum or starter material that needs to be added to the biogas digester system. Experiments conducted in the digester

Determination of succession of rumen bacterial species in nursing beef calves

Science.gov (United States)

Ruminants are typically born with a non-functional rumen essentially devoid of microorganisms. The succession of the microbial population in the rumen from birth to animal maturity is of interest due to the key role that the rumen microbial population plays in the overall health and productivity of ...

Effects of rumen-escape starch and coarseness of ingredients in pelleted concentrates on performance and rumen wall characteristics of rosé veal calves.

Science.gov (United States)

Vestergaard, M; Jarltoft, T C; Kristensen, N B; Børsting, C F

2013-08-01

The objective was to study the effect of rumen-escape starch and coarseness of ingredients in pelleted concentrates on performance, carcass quality and rumen wall characteristics in rosé veal calf production. Two alternative concentrates (Coarse and Slow) were compared with a traditional (Control) concentrate. Control was based on finely ground ingredients, whereas in Coarse, the same ingredients were coarsely ground resulting in a mean particle size before pelleting of 1.5 in Coarse and 0.6 mm in Control. Slow compared with Control and Coarse contained finely ground sorghum and corn instead of barley and wheat which increased the amount of rumen-escape starch to 59 compared with 22 g/kg in Control and Coarse. All concentrates had the same total starch (362 g/kg), NDF (168 g/kg), CP (154 g/kg) and DE (15.5 MJ/kg DM) content and a pellet diameter of 3.5 to 4 mm. Use of an 'indicator of starch digestibility' method gave a value of 98.6% for Control and Coarse and 91.1% for Slow (P 0.05). Papillae length and shape evaluated in atrium ruminis and the cranial part of the ventral rumen sac at slaughter were not affected by type of concentrate (P > 0.05). Rumen wall characteristics showed degrees of plaque formation (i.e., papillary aggregation), hyperaemia and necrotic areas in all treatment groups, but with no general difference between type of concentrate (P > 0.05). Incidence of liver abscesses (LAs, 16%) was not affected by type of concentrate (P > 0.05). There were no differences in performance or rumen wall characteristics between liver-abscessed and non-abscessed calves. The results show a high level of production performance with the three types of pelleted concentrates and indicates that neither the more coarse ingredients nor the additional rumen-escape starch tested, when fed ad libitum, could improve rumen wall characteristics or reduce LAs of rosé veal calves.

Effect of sugar fatty acid esters on rumen fermentation in vitro.

Science.gov (United States)

Wakita, M; Hoshino, S

1987-11-01

1. The effect of sugar fatty acid esters (SFEs; currently used as food additives for human consumption) on rumen volatile fatty acids (VFA) and gas production was studied with sheep rumen contents in vitro. 2. Some SFEs having monoester contents of more than 70% increased the molar proportion of propionate in conjunction with reduction in the acetate:propionate ratio when the individual SFE was added to rumen contents in a final concentration of 4 g/l. Laurate sugar ester was the most potent propionate enhancer and rumen gas depressor, the effective dose being as low as 1 g/l in a final concentration. Fatty acid esters other than SFEs had little, if any, effect on rumen VFA production and their molar proportions. 3. Approximately 50% of laurate sugar ester was hydrolysed by in vitro incubation with rumen fluid for 2 h. The addition of fatty acids and sucrose was also effective in the alterations of rumen VFA and gas production. However, the effect of SFEs on in vitro rumen fermentation was significantly greater than that of their constituent fatty acids or sucrose, or both. Accordingly, the effect appeared to be ascribed to the complex action of SFE itself and to its constituents, free fatty acids and sucrose. 4. SFEs, at the level of 4 g/l, reduced substantially the froth formation (ingesta volume increase) and seemed to be effective for the prevention of bloat.

MHC molecules protect T cell epitopes against proteolytic destruction

DEFF Research Database (Denmark)

Mouritsen, S; Meldal, M; Werdelin, O

1992-01-01

There is a subtle duality in the role of proteolytic enzymes in Ag processing. They are required to fragment protein Ag ingested by APC. However, prolonged exposure to proteolytic enzymes may lead to a complete degradation of the Ag, leaving nothing for the T cell system to recognize. What ensures...

Similarity of the ruminal bacteria across individual lactating cows.

Science.gov (United States)

Jami, Elie; Mizrahi, Itzhak

2012-06-01

Dairy cattle hold enormous significance for man as a source of milk and meat. Their remarkable ability to convert indigestible plant mass into these digestible food products resides in the rumen - an anaerobic chambered compartment - in the bovine digestive system. The rumen houses a complex microbiota which is responsible for the degradation of plant material, consequently enabling the conversion of plant fibers into milk and meat and determining their quality and quantity. Hence, an understanding of this complex ecosystem has major economic implications. One important question that is yet to be addressed is the degree of conservation of rumen microbial composition across individual animals. Here we quantified the degree of similarity between rumen bacterial populations of 16 individual cows. We used real-time PCR to determine the variance of specific ruminal bacterial species with different metabolic functions, revealing that while some bacterial strains vary greatly across animals, others show only very low variability. This variance could not be linked to the metabolic traits of these bacteria. We examined the degree of similarity in the dominant bacterial populations across all animals using automated ribosomal intergenic spacer analysis (ARISA), and identified a bacterial community consisting of 32% operational taxonomic units (OTUs) shared by at least 90% of the animals and 19% OTUs shared by 100% of the animals. Looking only at the presence or absence of each OTU gave an average similarity of 75% between each cow pair. When abundance of each OTU was added to the analysis, this similarity decreased to an average of less than 60%. Thus, as suggested in similar recent studies of the human gut, a bovine rumen core microbiome does exist, but taxa abundance may vary greatly across animals. Copyright © 2012 Elsevier Ltd. All rights reserved.

Feeding Aspergillus oryzae Fermentation Culture (AOFC to Growing Sheep: 1. The Effect of AOFC on Rumen Fermentation

Directory of Open Access Journals (Sweden)

Darwinsyah Lubis

2002-10-01

Full Text Available Cultures of fungi, especially Aspergillus oryzae, have been of interest to animal nutritionists to increase feed efficiency. Many experiments have been done and showed positive results on rumen fermentation and productivity of ruminants. This paper reports the results of an in vivo study on feeding Aspergillus oryzae, fermentation culture (AOFC to growing sheep. ‘Onggok’ (tapioca processing waste was used as media for AO cultivation after being enriched with a mineral mixture. Commercial concentrate (GT-03 was fed to 15 growing sheep supplemented with 0% (C0, 5% (C1, and 10% (C2 AOFC (w/w. Chopped fresh King grass was used as a basal diet. The 3 treatments were randomly allotted to the sheep according to randomized block design with 5 replications. The study was carried out for 14 weeks. Digestion trial was conducted in the last 10 days of experiment. All feed and fecal samples were analyzed for nutrients. Rumen fluid was sampled at the mid experimental period. Analyses were done on rumen pH, ammonia content, (VFA volatile fatly acids concentration, and also total digestive tract digestibility of dry and organic matter, crude protein, and total fiber (NDF. Differences in treatment means were analyzed by Duncan’s MRT. Feeding AOFC resulted in increased (P<0.05 digestibility of crude protein from 59.6% in control sheep to 65.5% in sheep fed concentrate with 10% AOFC supplementation. The same pattern also occurred for NDF, but no effect was found on dry and organic matter. Higher fiber digestibility with AOFC supplementation was in line with an increase (P<0.05 in cellulolytic bacteria population in the rumen. VFA produced also increased (P<0.05, as well as individual acids content, primarily acetate and propionate. No differences (P<0.05 were detected in rumen pH and ammonia content. It appears that AOFC is more suitable for the purpose of meat production.

Research Applications of Proteolytic Enzymes in Molecular Biology

OpenAIRE

Mótyán, János András; Tóth, Ferenc; Tőzsér, József

2013-01-01

Proteolytic enzymes (also termed peptidases, proteases and proteinases) are capable of hydrolyzing peptide bonds in proteins. They can be found in all living organisms, from viruses to animals and humans. Proteolytic enzymes have great medical and pharmaceutical importance due to their key role in biological processes and in the life-cycle of many pathogens. Proteases are extensively applied enzymes in several sectors of industry and biotechnology, furthermore, numerous research applications ...

Title: Effects of supplementing humic/fulvic acid on rumen ...

African Journals Online (AJOL)

Casey McMurphy

Effects of supplementing humates on rumen fermentation in Holstein ... research on the utilization of humates in beef cattle diets and their effect on rumen fermentation. .... potassium chloride, 80 g/kg; magnesium oxide, 34.5 g/kg; ammonium ...

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

Science.gov (United States)

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 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 and between microbial domains due to greater competition for substrates that are of limited availability as a result of biofilm formation.

Entry of blood urea into the rumen of the llama

International Nuclear Information System (INIS)

Hinderer, S.; Engelhardt, W. von

1976-01-01

Llamas were provided with a large rumen fistula, and the transfer of blood urea into the temporarily isolated rumen, cleaned and filled with test solution was measured. Plasma urea clearance due to transfer of blood urea across the rumen wall should indicate changes in its permeability to urea. Clearance values were highest with CO 2 or with high concentrations of butyric acid. Permeability was low when food was with-held and when no volatile fatty acids were present in the solution. The permeability of the rumen wall to blood urea can be altered significantly. These changes can affect blood urea transfer more extensively than changes in the plasma urea concentration within physiological ranges

Appearance and dynamics of rumen motility in newborn calves

International Nuclear Information System (INIS)

Kostov, Y.; Aleksandrova, V.

2010-01-01

The appearance and dynamics of rumen motility in newborn calves were studied by means of radiotelemetry. Rumen contractions were registered right after birth. Their amplitude was growing gradually and that was observed best in the first month after birth

In vitro effects of sodium bicarbonate buffer on rumen fermentation, levels of lipopolysaccharide and biogenic amine, and composition of rumen microbiota.

Science.gov (United States)

Mao, Shengyong; Huo, Wenjie; Liu, Junhua; Zhang, Ruiyang; Zhu, Weiyun

2017-03-01

Diets containing high levels of carbohydrates provoke a rapid decrease of rumen pH and high levels of biogenic amines and lipopolysaccharides (LPS), which severely impair the health and performance of ruminants. The goal of this study was to evaluate the effects of sodium bicarbonate (BC) buffer on rumen fermentation, levels of LPS and biogenic amine, and composition of rumen microbiota using in vitro rumen cultures. Sodium bicarbonate supplementation increased (P < 0.05) the final pH levels and concentrations of total volatile fatty acids and LPS, as well as the proportions of acetate, propionate, isobutyrate, isovalerate and valerate, and it decreased (P < 0.05) the proportion of butyrate and the levels of lactic acid, methylamine, tryptamine, tyramine, histamine and putrescine compared with the control. Pyrosequencing of the 16S rRNA gene showed that BC inclusion increased (P < 0.05) the bacterial diversity index compared with the control. Adding BC also decreased (P < 0.05) the relative abundance of Streptococcus and Butyrivibrio and increased (P < 0.05) the proportions of Ruminococcus, Succinivibrio and Prevotella. Sodium bicarbonate supplementation has beneficial effects in the reduction of bioamine levels and the increase in ruminal pH, and in modifying the microbial ecology of the rumen; however, it results in an accumulation of LPS under high-grain diet conditions. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

The effect of an isoflavonid-rich liquorice extract on fermentation, methanogenesis and the microbiome in the rumen simulation technique.

Science.gov (United States)

Ramos-Morales, E; Rossi, G; Cattin, M; Jones, E; Braganca, R; Newbold, C J

2018-03-01

Due to the antimicrobial activity of flavonoids, it has been suggested that they may provide a possible alternative to antibiotics to stimulate productivity and reduce the environmental load of ruminant agriculture. We hypothesised that an extract of liquorice, rich in prenylated isoflavonoids and particularly glabridin, might potentially improve the efficiency of nitrogen utilisation and reduce methane production in the rumen. When added to a long-term rumen simulating fermentor (RUSITEC), liquorice extract at 1 g L-1 decreased ammonia production (-51%; P fermentation process. When added at 2 g L-1, decreases in not only ammonia production (-77%; P fermentation were probably related to a decrease in protozoa numbers, a less diverse bacteria population as well as changes in the structure of both the bacterial and archaeal communities. The inclusion of an isoflavonoid-rich extract from liquorice in the diet may potentially improve the efficiency of the feed utilisation by ruminants.

Rumen bacterial community evaluated by 454 pyrosequencing and terminal restriction fragment length polymorphism analyses in dairy sheep fed marine algae.

Science.gov (United States)

Castro-Carrera, T; Toral, P G; Frutos, P; McEwan, N R; Hervás, G; Abecia, L; Pinloche, E; Girdwood, S E; Belenguer, A

2014-03-01

Developing novel strategies to increase the content of bioactive unsaturated fatty acids (FA) in ruminant-derived products requires a deeper understanding of rumen biohydrogenation and bacteria involved in this process. Although high-throughput pyrosequencing may allow for a great coverage of bacterial diversity, it has hardly been used to investigate the microbiology of ruminal FA metabolism. In this experiment, 454 pyrosequencing and a molecular fingerprinting technique (terminal restriction fragment length polymorphism; T-RFLP) were used concurrently to assess the effect of diet supplementation with marine algae (MA) on the rumen bacterial community of dairy sheep. Eleven lactating ewes were divided in 2 lots and offered a total mixed ration based on alfalfa hay and concentrate (40:60), supplemented with 0 (control) or 8 (MA) g of MA/kg of dry matter. After 54 d on treatments, animals were slaughtered and samples of rumen content and fluid were collected separately for microbial analysis. Pyrosequencing yielded a greater coverage of bacterial diversity than T-RFLP and allowed the identification of low abundant populations. Conversely, both molecular approaches pointed to similar conclusions and showed that relevant changes due to MA addition were observed within the major ruminal phyla, namely Bacteroidetes, Firmicutes, and Proteobacteria. Decreases in the abundance of unclassified Bacteroidales, Porphyromonadaceae, and Ruminococcaceae and increases in as-yet uncultured species of the family Succinivibrionaceae, might be related to a potential role of these groups in different pathways of rumen FA metabolism. Diet supplementation with MA, however, had no effect on the relative abundance of Butyrivibrio and Pseudobutyrivibrio genera. In addition, results from both 454 pyrosequencing and T-RFLP indicate that the effect of MA was rather consistent in rumen content or fluid samples, despite inherent differences between these fractions in their bacterial composition

Manipulation of the rumen to increase ruminant production

International Nuclear Information System (INIS)

Nolan, J.V.; Leng, R.A.

1989-01-01

Manipulation of the rumen should be undertaken with a view to optimizing the supply of specific nutrients to the host animal. Especially important are the volatile fatty acids (glucogenic and non-glucogenic), and dietary and bypass amino acids and lipids. The animal's requirement for these nutrients varies according to its physiological state (growing, pregnant or lactating), its health (disease or parasite load), previous dietary history and also the prevailing climatic conditions. Often the availability of total protein (amino acids) relative to oxidizable substrates (the protein/energy or P/E ratio) is the primary limitation to voluntary feed intake and to efficient use of absorbed nutrients. In these instances, the first objective should be to maximize the yield of microbial amino acids from the rumen by manipulations that increase the net efficiency of microbial synthesis, e.g. by altering methods of feed processing, supplementing with urea and minerals, or eliminating protozoa. Further improvements in the P/E ratio can be attained by supplementing the animal with a dietary protein source with good rumen bypass characteristics in order to provide additional amino acids for intestinal absorption. Other potential manipulations of the rumen include the use of chemicals to modify rumen fermentation patterns and supplementation with long chain fatty acids (LCFA) (as a dense source of energy supplying substrate) in rumen-inert forms (e.g. as Ca-LCFA). However, when energy dense dietary supplements are given, it may be necessary to include a bypass protein source to maintain the P/E ratio. Novel possibilities for manipulation include the use of molecular biology techniques to create microorganisms with enhanced production of enzymes that promote degradation of structural carbohydrates, or with an ability to secrete chemicals that are toxic to protozoa. (author). 68 refs, 4 figs, 1 tab

Effects of nitrate addition to a diet on fermentation and microbial populations in the rumen of goats, with special reference to Selenomonas ruminantium having the ability to reduce nitrate and nitrite.

Science.gov (United States)

Asanuma, Narito; Yokoyama, Shota; Hino, Tsuneo

2015-04-01

This study investigated the effects of dietary nitrate addition on ruminal fermentation characteristics and microbial populations in goats. The involvement of Selenomonas ruminantium in nitrate and nitrite reduction in the rumen was also examined. As the result of nitrate feeding, the total concentration of ruminal volatile fatty acids decreased, whereas the acetate : propionate ratio and the concentrations of ammonia and lactate increased. Populations of methanogens, protozoa and fungi, as estimated by real-time PCR, were greatly decreased as a result of nitrate inclusion in the diet. There was modest or little impact of nitrate on the populations of prevailing species or genus of bacteria in the rumen, whereas Streptococcus bovis and S. ruminantium significantly increased. Both the activities of nitrate reductase (NaR) and nitrite reductase (NiR) per total mass of ruminal bacteria were increased by nitrate feeding. Quantification of the genes encoding NaR and NiR by real-time PCR with primers specific for S. ruminantium showed that these genes were increased by feeding nitrate, suggesting that the growth of nitrate- and nitrite-reducing S. ruminantium is stimulated by nitrate addition. Thus, S. ruminantium is likely to play a major role in nitrate and nitrite reduction in the rumen. © 2014 Japanese Society of Animal Science.

Rumen management during aphagia : review article

Directory of Open Access Journals (Sweden)

A.S. Shakespeare

2008-05-01

Full Text Available 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.

Changes in microbial and nutrient composition associated with rumen content compost incubation.

Science.gov (United States)

Shrestha, Karuna; Shrestha, Pramod; Adetutu, Eric M; Walsh, Kerry B; Harrower, Keith M; Ball, Andrew S; Midmore, David J

2011-02-01

Physico-chemical and microbiological investigations were carried out on rumen content material composted for nine months, fresh vermicasts (obtained after passing the same compost through the guts of a mixture of three species of earthworms: Eisenia fetida, Lumbricus rubellus and Perionyx excavates) and microbially enhanced extracts derived from rumen compost, vermicast and vermicast leachate incubated for up to 48 h. Compared to composted rumen contents, vermicast was only improved in terms of microbial biomass C, while vermicast leached extract was significantly higher in NH(4)(+)-N,PO(4)(-)-P, humic acid, bacterial counts and total microbial activity compared to rumen compost extract. Although no difference between treatments was observed in genetic diversity as indicated by DGGE analysis, community level functional diversity of vermicast leached extract (Biolog™) was higher than that of composted rumen contents, vermicast and rumen compost extract indicating an enhancement of microbial activity rather than diversity due to liquid incubation. Copyright © 2010 Elsevier Ltd. All rights reserved.

Differential carbohydrate media and anaerobic replica plating techniques in delineating carbohydrate-utilizing subgroups in rumen bacterial populations.

OpenAIRE

Leedle, J A; Hespell, R B

1980-01-01

A basal (BC) medium devoid of added carbohydrates, a complete (CC) medium containing nine carbohydrates were developed for enumerating rumen bacteria. The colony counts on the BC medium were 85 to 100% of those obtained on the CC medium. These colonies were pinpoint size (less than or equal to mm in diameter) but increased in size (2 to 5 mm in diameter) when carbohydrates were subsequently added. With the CC medium or other media tested, the colony counts were 20 to 50% higher on plates than...

Evaluation of the effect of fat content of sunflower meal on rumen ...

African Journals Online (AJOL)

Yomi

2012-01-03

Jan 3, 2012 ... addition of yellow grease and corn oil increased in vitro rumen degradation and gas production. ... Culturing of rumen anaerobic fungi. Rumen anaerobic fungi were isolated from wheat straw residues, ... concentrate : forage diet (corn grain, barley grain and wheat bran : sugarcane silage, corn silage, alfalfa ...

Interaction of Leptospira interrogans with Human Proteolytic Systems Enhances Dissemination through Endothelial Cells and Protease Levels

Science.gov (United States)

Vieira, Monica L.; Alvarez-Flores, Miryam P.; Kirchgatter, Karin; Romero, Eliete C.; Alves, Ivy J.; de Morais, Zenaide M.; Vasconcellos, Silvio A.; Chudzinski-Tavassi, Ana M.

2013-01-01

We have recently reported the ability of Leptospira to capture plasminogen (PLG) and generate plasmin (PLA) bound on the microbial surface in the presence of exogenous activators. In this work, we examined the effects of leptospiral PLG binding for active penetration through the endothelial cell barrier and activation. The results indicate that leptospires with PLG association or PLA activation have enhanced migration activity through human umbilical vein endothelial cell (HUVEC) monolayers compared with untreated bacteria. Leptospira cells coated with PLG were capable of stimulating the expression of PLG activators by HUVECs. Moreover, leptospires endowed with PLG or PLA promoted transcriptional upregulation matrix metalloprotease 9 (MMP-9). Serum samples from patients with confirmed leptospirosis showed higher levels of PLG activators and total MMP-9 than serum samples from normal (healthy) subjects. The highest level of PLG activators and total MMP-9 was detected with microscopic agglutination test (MAT)-negative serum samples, suggesting that this proteolytic activity stimulation occurs at the early stage of the disease. Furthermore, a gelatin zymography profile obtained for MMPs with serum samples from patients with leptospirosis appears to be specific to leptospiral infection because serum samples from patients with unrelated infectious diseases produced no similar degradation bands. Altogether, the data suggest that the Leptospira-associated PLG or PLA might represent a mechanism that contributes to bacterial penetration of endothelial cells through an activation cascade of events that enhances the proteolytic capability of the organism. To our knowledge, this is the first proteolytic activity associated with leptospiral pathogenesis described to date. PMID:23478319

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.

Science.gov (United States)

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.

Effects of rumen-escape starch and coarseness of ingredients in pelleted concentrates on performance and rumen wall characteristics of rosé veal calves

DEFF Research Database (Denmark)

Vestergaard, Mogens; Jarltoft, Terese Christel; Kristensen, Niels Bastian

2013-01-01

The objective was to study the effect of rumen-escape starch and coarseness of ingredients in pelleted concentrates on performance, carcass quality and rumen wall characteristics in rosé veal calf production. Two alternative concentrates (Coarse and Slow) were compared with a traditional (Control...

Increasing Biogas Production Rate from Cattle Manure Using Rumen Fluid as Inoculums

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Budiyono Budiyono

2013-10-01

Full Text Available Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 In this study, rumen fluid of animal ruminant was used as inoculums to increase biogas production rate from cattle manure at mesophilic condition. A series of laboratory experiments using 400 ml biodigester were performed in batch operation mode. Given 100 grams of fresh cattle manure (M was fed to each biodigester and mixed with rumen fluid (R and tap water (W in several ratio resulting six different M:W:R ratio contents i.e. 1:1:0; 1:0.75:0.25; 1:0.5:0.5; 1:0.25:0.75; and 1:0:1 (correspond to 0; 12.5; 25, 37.5; 50, and 100 % rumen, respectively and six different total solid (TS contents i.e. 2.6, 4.6, 6.2, 7.4, 9.2, 12.3, and 18.4 %. The operating temperatures were at room temperature. The results showed that the rumen fluid inoculated to biodigester significantly effected the biogas production. Rumen fluid inoculums caused biogas production rate and efficiency increase more than two times in compare to manure substrate without rumen fluid inoculums. The best performance for biogas production was the digester with rumen fluid and TS content in the range of 25-50 % and 7.4 and 9.2 %, respectively. These results suggest that, based on TS content effects to biogas yield, rumen fluid inoculums exhibit the similar effect with other inoculums. Increasing rumen content will also increase biogas production. Due to the optimum total solid (TS content for biogas production between 7-9 % (or correspond to more and less manure and total liquid 1:1, the rumen fluid content of 50 % will give the best performance for biogas production. The future work will be carried out to study the dynamics of biogas production if both the rumen fluid inoculums and manure are fed in the continuous system Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 Doi: 10.12777/ijse.6.1.31-38 [How to cite this article: Budiyono, Widiasa, I.N., Johari, S. and Sunarso. (2014. Increasing Biogas

Variation among Dairy Cows in Rumen Liquid Fermentation Characteristics

DEFF Research Database (Denmark)

Zhu, Zhigang; Kristensen, Lise; Højbjerg, Ole

Volatile fatty acids are the main energy product from rumen fermentation. This study investigated the individuality of VFA concentrations in samples of rumen fluid obtained from 10 Holstein cows using a esophageal probe to take samples repeatedly over a 7 week period. Systematic changes were seen...

The position of rumenic acid on triacylglycerols alters its bioavailability in rats.

Science.gov (United States)

Chardigny, J M; Masson, E; Sergiel, J P; Darbois, M; Loreau, O; Noël, J P; Sébédio, J-L

2003-12-01

The metabolic fate of rumenic acid (9cis,11trans-octadecenoic acid) related to its position on the glycerol moiety has not yet been studied. In the present work, synthetic triacylglycerols (TAG) esterified with oleic and rumenic acids were prepared. Rats were force-fed synthetic dioleyl monorumenyl glycerol with (14)C labeled rumenic acid in the internal (sn-2) or in the external position (sn-1 or sn-3). Rats were then placed in metabolic cages for 16 h. At the end of the experiment, the radioactivity in tissues, carcass and expired CO(2) was measured. Rumenic acid that was esterified at the external positions on the TAG was better absorbed and oxidized to a greater extent than when esterified at the internal position. The fatty acid from the 2-TAG form was also better incorporated into the rat carcass. In the liver, rumenic acid appeared mainly in TAG (50%) and to a lesser extent in phospholipids (33%) whatever its dietary form. Moreover, analyses of lipids from Camembert cheese and butter revealed that rumenic acid was located mainly on the sn-1 or sn-3 positions (74%). Taken together, these data suggest that rumenic acid from dairy fat may be well absorbed and used extensively for energy production.

Effects of early feeding on the host rumen transcriptome and bacterial diversity in lambs

Science.gov (United States)

Wang, Weimin; Li, Chong; Li, Fadi; Wang, Xiaojuan; Zhang, Xiaoxue; Liu, Ting; Nian, Fang; Yue, Xiangpeng; Li, Fei; Pan, Xiangyu; La, Yongfu; Mo, Futao; Wang, Fangbin; Li, Baosheng

2016-01-01

Early consumption of starter feed promotes rumen development in lambs. We examined rumen development in lambs fed starter feed for 5 weeks using histological and biochemical analyses and by performing high-throughput sequencing in rumen tissues. Additionally, rumen contents of starter feed-fed lambs were compared to those of breast milk-fed controls. Our physiological and biochemical findings revealed that early starter consumption facilitated rumen development, changed the pattern of ruminal fermentation, and increased the amylase and carboxymethylcellulase activities of rumen micro-organisms. RNA-seq analysis revealed 225 differentially expressed genes between the rumens of breast milk- and starter feed-fed lambs. These DEGs were involved in many metabolic pathways, particularly lipid and carbohydrate metabolism, and included HMGCL and HMGCS2. Sequencing analysis of 16S rRNA genes revealed that ruminal bacterial communities were more diverse in breast milk-than in starter feed-fed lambs, and each group had a distinct microbiota. We conclude that early starter feeding is beneficial to rumen development and physiological function in lambs. The underlying mechanism may involve the stimulation of ruminal ketogenesis and butanoate metabolism via HMGCL and HMGCS2 combined with changes in the fermentation type induced by ruminal microbiota. Overall, this study provides insights into the molecular mechanisms of rumen development in sheep. PMID:27576848

Effect of partially replacing a barley-based concentrate with flaxseed-based products on the rumen bacterial population of lactating Holstein dairy cows.

Science.gov (United States)

Castillo-Lopez, E; Moats, J; Aluthge, N D; Ramirez Ramirez, H A; Christensen, D A; Mutsvangwa, T; Penner, G B; Fernando, S C

2018-01-01

The effects of partial replacement of a barley-based concentrate with flaxseed-based products on the rumen bacterial population of lactating Holstein dairy cows were evaluated. Treatments fed were CONT, a normal diet that included barley silage, alfalfa hay and a barley-based concentrate that contained no flaxseed or faba beans; FLAX, inclusion of a nonextruded flaxseed-based product containing 55·0% flaxseed, 37·8% field peas and 6·9% alfalfa; EXT, similar to FLAX, but the product was extruded and EXTT, similar to FLAX, but product was extruded and field peas were replaced by high-tannin faba beans. The rumen bacterial population was evaluated by utilizing 16S rRNA gene sequencing. Most abundant phyla, families and genera were unaffected. However, some taxa were affected; for example, unsaturated fatty acid content was negatively correlated with Clostridiaceae, and tannin content was negatively correlated with BS11 and Paraprevotellaceae. Predominant rumen bacterial taxa were not affected, but the abundance of some taxa found in lower proportions shifted, possibly due to sensitivity to unsaturated fatty acids or tannins. Flaxseed-based products were effective for partially replacing barley-based concentrate in rations of lactating dairy cows. No negative effects of these products were observed on the abundance of predominant rumen bacterial taxa, with only minor shifts in less abundant bacteria. © 2017 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.

Isolation of chitinolytic Clostridium sp. NCR from Mehsani buffalo rumen, its genomic analysis and potential role in rumen

OpenAIRE

Nathani, Neelam M.; Duggirala, Srinivas M.; M., Chandra Shekar; Kothari, Ramesh K.; Joshi, Chaitanya G.

2015-01-01

Genomic analysis of Clostridium sp. NCR, an anaerobic Gram positive bacterium which was isolated from rumen fluid of Mehsani breed of buffalo revealed presence of various environmental gene tags (EGTs) involved in pathways for utilizing a wide range of substrates. Here we report the sequence of this rumen isolate, its whole genome sequence has been deposited in DDBJ/EMBL/GenBank under the accession number JQHY00000000. The genome comprises of a 3.62-Mb draft genome with a G + C content of 28....

Proteolytic crosstalk in multi-protease networks

Science.gov (United States)

Ogle, Curtis T.; Mather, William H.

2016-04-01

Processive proteases, such as ClpXP in E. coli, are conserved enzyme assemblies that can recognize and rapidly degrade proteins. These proteases are used for a number of purposes, including degrading mistranslated proteins and controlling cellular stress response. However, proteolytic machinery within the cell is limited in capacity and can lead to a bottleneck in protein degradation, whereby many proteins compete (‘queue’) for proteolytic resources. Previous work has demonstrated that such queueing can lead to pronounced statistical relationships between different protein counts when proteins compete for a single common protease. However, real cells contain many different proteases, e.g. ClpXP, ClpAP, and Lon in E. coli, and it is not clear how competition between proteins for multiple classes of protease would influence the dynamics of cellular networks. In the present work, we theoretically demonstrate that a multi-protease proteolytic bottleneck can substantially couple the dynamics for both simple and complex (oscillatory) networks, even between substrates with substantially different affinities for protease. For these networks, queueing often leads to strong positive correlations between protein counts, and these correlations are strongest near the queueing theoretic point of balance. Furthermore, we find that the qualitative behavior of these networks depends on the relative size of the absolute affinity of substrate to protease compared to the cross affinity of substrate to protease, leading in certain regimes to priority queue statistics.

Morphological studies on rumen development in West African Dwarf ...

African Journals Online (AJOL)

We studied the gross and light microscopic structures of rumen in fetal, neonatal and adult West African Dwarf (WAD) goats obtained from Nsukka and Igboeze South Local Government Areas (L.G.A) of Enugu State. After euthanasia the rumen was ligated, dissected out and the volume determined by flotation and ...

Influence of phenolic compounds on rumen microbial activity

International Nuclear Information System (INIS)

Vitti, D.M.S.S.; Abdalla, A.L.; Silva Filho, J.C.

1985-01-01

An 'in vitro' experiment is carried out to examine the effect of tannic acid on rumen microbial activity, due to the toxicity of phenolic acids on many microrganisms. Rumen content is incubated with sodium bicarbonate, glucose and different quantities of tannic acid. 1 μCi of 32 p-labelled phosphate is added and after 6 hours the incorporated activity is measured. (M.A.C.) [pt

Rumen degradability of some feed legume seeds

OpenAIRE

González , Javier; Andrés , Santiago

2003-01-01

International audience; The aim of this work was to determine the effective degradability (ED) of CP for different feed legume seeds and the possible relationship with their physical and chemical characteristics. The ED was measured using nylon bags and rumen outflow rate techniques on three rumen cannulated wethers fed at 40 g DM$\\cdot$kg$^{-0.75}$, with a 2:1 (on DM basis) hay to concentrate diet. Nine seed samples of the following legume species were tested: lupin (Lupinus albus L., cultiv...

Fermentation of Ammonia Fiber Expansion Treated and Untreated Barley Straw in a Rumen Simulation Technique Using Rumen Inoculum from Cattle with Slow versus Fast Rate of Fiber Disappearance.

Science.gov (United States)

Griffith, Candace L; Ribeiro, Gabriel O; Oba, Masahito; McAllister, Tim A; Beauchemin, Karen A

2016-01-01

The purpose of this study was to determine the effect of rumen inoculum from heifers with fast vs. slow rate of in situ fiber digestion on the fermentation of complex versus easily digested fiber sources in the forms of untreated and Ammonia Fiber Expansion (AFEX) treated barley straw, respectively, using an artificial rumen simulation technique (Rusitec). In situ fiber digestion was measured in a previous study by incubating untreated barley straw in the rumen of 16 heifers fed a diet consisting of 700 g/kg barley straw and 300 g/kg concentrate. The two heifers with fastest rate of digestion (Fast ≥ 4.18% h -1 ) and the two heifers with the slowest rate of digestion (Slow ≤ 3.17% h -1 ) were chosen as inoculum donors for this study. Two Rusitec apparatuses each equipped with eight fermenters were used in a completely randomized block design with two blocks (apparatus) and four treatments in a 2 × 2 factorial arrangement of treatments (Fast or Slow rumen inoculum and untreated or AFEX treated straw). Fast rumen inoculum and AFEX straw both increased ( P 0.05) methane production per gram of digested material for both untreated and AFEX straw, and reduced (interaction, P < 0.05) acetate: propionate ratio for untreated straw. Greater relative populations of Ruminococcus albus ( P < 0.05) and increased microbial N production ( P = 0.045) were observed in Fast rumen inoculum. AFEX straw in Fast inoculum had greater total bacterial populations than Slow, but for untreated straw this result was reversed (interaction, P = 0.013). These findings indicate that differences in microbial populations in rumen fluid contribute to differences in the capacity of rumen inoculum to digest fiber.

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

Science.gov (United States)

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...

Sirtuins and Proteolytic Systems: Implications for Pathogenesis of Synucleinopathies

Directory of Open Access Journals (Sweden)

Belém Sampaio-Marques

2015-05-01

Full Text Available Insoluble and fibrillar forms of α-synuclein are the major components of Lewy bodies, a hallmark of several sporadic and inherited neurodegenerative diseases known as synucleinopathies. α-Synuclein is a natural unfolded and aggregation-prone protein that can be degraded by the ubiquitin-proteasomal system and the lysosomal degradation pathways. α-Synuclein is a target of the main cellular proteolytic systems, but it is also able to alter their function further, contributing to the progression of neurodegeneration. Aging, a major risk for synucleinopathies, is associated with a decrease activity of the proteolytic systems, further aggravating this toxic looping cycle. Here, the current literature on the basic aspects of the routes for α-synuclein clearance, as well as the consequences of the proteolytic systems collapse, will be discussed. Finally, particular focus will be given to the sirtuins’s role on proteostasis regulation, since their modulation emerged as a promising therapeutic strategy to rescue cells from α-synuclein toxicity. The controversial reports on the potential role of sirtuins in the degradation of α-synuclein will be discussed. Connection between sirtuins and proteolytic systems is definitely worth of further studies to increase the knowledge that will allow its proper exploration as new avenue to fight synucleinopathies.

In Vitro Digestibilities of Six Rumen Protected Fat-Protein Supplement Formulas

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Lilis Hartati

2012-01-01

Full Text Available Abstract. The aim of the research was to evaluate the efficacy of protection method of rumen protected fat-protein supplements. In vitro digestibility test was carried out to examine nutrients digestibility of different supplement formula based on the sources of protein and oil. The research used two sources of fat namely crude palm oil (CPO and fish oil (FO and three sources of protein namely milk skim, soy flour, and soybean meal. Thus there were 6 combinations that subjected in the in vitro digestibility test. The observed variables were the digestibility of dry matter (DM, organic matter (OM, crude fat (CF, and crude protein (CP. Results indicated that the method for protecting protein and fat was effective. This was showed by low nutrients digestibility in the rumen and high nutrients digestibility in the post rumen. In conclusion the combination between skim milk and CPO gave the best results among the other supplement formula. Keywords: rumen protected nutrient, fat-protein supplement, rumen digestibility, in vitro Animal Production 14(1:1-5, January 2012

Effect of culturing conditions on the expression of key enzymes in the proteolytic system of Lactobacillus bulgaricus *

Science.gov (United States)

Hou, Jun-cai; Liu, Fei; Ren, Da-xi; Han, Wei-wei; Du, Yue-ou

2015-01-01

The proteolytic system of Lactobacillus bulgaricus breaks down milk proteins into peptides and amino acids, which are essential for the growth of the bacteria. The aim of this study was to determine the expressions of seven key genes in the proteolytic system under different culturing conditions (different phases, initial pH values, temperatures, and nitrogen sources) using real-time polymerase chain reaction (RT-PCR). The transcriptions of the seven genes were reduced by 30-fold on average in the stationary phase compared with the exponential growth phase. The transcriptions of the seven genes were reduced by 62.5-, 15.0-, and 59.0-fold in the strains KLDS 08006, KLDS 08007, and KLDS 08012, respectively, indicating that the expressions of the seven genes were significantly different among strains. In addition, the expressions of the seven genes were repressed in the MRS medium containing casein peptone. The effect of peptone supply on PepX transcription was the weakest compared with the other six genes, and the impact on OppD transcription was the strongest. Moreover, the expressions of the seven genes were significantly different among different strains (PLactobacillus bulgaricus at the transcription level. PMID:25845365

Effect of culturing conditions on the expression of key enzymes in the proteolytic system of Lactobacillus bulgaricus.

Science.gov (United States)

Hou, Jun-cai; Liu, Fei; Ren, Da-xi; Han, Wei-wei; Du, Yue-ou

2015-04-01

The proteolytic system of Lactobacillus bulgaricus breaks down milk proteins into peptides and amino acids, which are essential for the growth of the bacteria. The aim of this study was to determine the expressions of seven key genes in the proteolytic system under different culturing conditions (different phases, initial pH values, temperatures, and nitrogen sources) using real-time polymerase chain reaction (RT-PCR). The transcriptions of the seven genes were reduced by 30-fold on average in the stationary phase compared with the exponential growth phase. The transcriptions of the seven genes were reduced by 62.5-, 15.0-, and 59.0-fold in the strains KLDS 08006, KLDS 08007, and KLDS 08012, respectively, indicating that the expressions of the seven genes were significantly different among strains. In addition, the expressions of the seven genes were repressed in the MRS medium containing casein peptone. The effect of peptone supply on PepX transcription was the weakest compared with the other six genes, and the impact on OppD transcription was the strongest. Moreover, the expressions of the seven genes were significantly different among different strains (Pproteolytic system genes in Lactobacillus bulgaricus at the transcription level.

The Effect of Selected Biostimulating Substance on the Degradation in the Rumen

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Eva Petrášková

2011-05-01

Full Text Available The cows with the canulla were divided into two groups - experimental and control. One cow was added to experimental product Biopolym FZT, second Biopolym granules. The samples of rumen fluid and feces were analyzed. The growth of microorganisms in the rumen fluid of experimental animals means the possibility of a positive impact of Biopolym on the degradation of feeding the rumen.

Rumen-protected choline: A significance effect on dairy cattle nutrition.

Science.gov (United States)

Jayaprakash, G; Sathiyabarathi, M; Robert, M Arokia; Tamilmani, T

2016-08-01

Choline is a vitamin-like substance it has multi-function in animal production, reproduction, and health. The transition period is most crucial stage in lactation cycle of dairy cows due to its association with negative hormonal and energy balances. Unfortunately, unprotected choline easily degrades in the rumen; therefore, choline added to the diet in a rumen-protected form. The use of rumen-protected choline (RPC) is a preventive measurement for the fatty liver syndrome and ketosis; may improve milk production as well as milk composition and reproduction parameters. This review summarizes the effectiveness of RPC on animal production, health, and reproduction.

The inhibitory effect of bovine rumen fluid on Salmonella typhimurium.

Science.gov (United States)

Chambers, P G; Lysons, R J

1979-05-01

The possible fate of Salmonella typhimurium in the rumen was investigated by monitoring rumen volatile fatty acids (VFA), lactate concentrations and pH over periods which included regular feeding and 48 h starvation. Preparations were made containing 50 per cent rumen fluid from the cow or VFA solutions, and then inoculated with S typhimurium. Viable counts before and after incubation for 24 h at 37 degrees C were compared. Incubation in broths with high concentrations of VFA and low pH resulted in a marked decrease in salmonella numbers, while lower VFA concentrations had little or no inhibitory effect on growth.

Determination of in vitro isoflavone degradation in rumen fluid.

Science.gov (United States)

Trnková, Andrea; Šancová, Kateřina; Zapletalová, Martina; Kašparovská, Jitka; Dadáková, Kateřina; Křížová, Ludmila; Lochman, Jan; Hadrová, Sylvie; Ihnatová, Ivana; Kašparovský, Tomáš

2018-06-01

The aim of this study was to determine the degradation of dietary isoflavones in rumen fluid under 2 feeding regimens. The experiments were performed in vitro using a rumen fluid buffer system. The rumen fluid was taken from cows fed either a hay diet or a concentrate-rich diet (the diet consisted of 34.6% maize silage, 17.6% haylage, 12.8% alfalfa hay, and 35.0% supplemental mixture on a dry matter basis). As a source of isoflavones, 40% soybean extract (Biomedica, Prague, Czech Republic) at levels of 5, 25, 50, and 75 mg per 40 mL of rumen fluid was used. Samples of soybean extract were incubated in triplicate at 39°C for 0, 3.0, 6.0, 12.0, and 24.0 h in incubation solution. The metabolism of daidzein and genistein was faster under concentrate-rich diet conditions. In general, production of equol started after 3 to 6 h of incubation and reached the highest rate after approximately 12 h of incubation regardless of the type of diet or concentration of extract. In most of the experiments, production of equol continued after 24 h of incubation. Generally, equol production was greater under the hay diet conditions. Furthermore, experiments with higher amounts of added soybean extract revealed possible inhibitory effects of high levels of isoflavones on the rumen microflora. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Fermentation of Ammonia Fiber Expansion Treated and Untreated Barley Straw in a Rumen Simulation Technique using Rumen Inoculum from Cattle with Slow Versus Fast Rate of Fiber Disappearance

Directory of Open Access Journals (Sweden)

Karen Ann Beauchemin

2016-11-01

Full Text Available The purpose of this study was to determine the effect of rumen inoculum from heifers with fast vs. slow rate of in situ fiber digestion on the fermentation of complex versus easily digested fiber sources in the forms of untreated and Ammonia Fiber Expansion (AFEX treated barley straw, respectively, using an artificial rumen simulation technique (Rusitec. In situ fiber digestion was measured in a previous study by incubating untreated barley straw in the rumen of sixteen heifers fed a diet consisting of 700 g/kg barley straw and 300 g/kg concentrate. The two heifers with fastest rate of digestion (Fast > 4.18 % h-1 and the two heifers with the slowest rate of digestion (Slow 0.05 methane per gram of digested material for both untreated and AFEX straw, and reduced (interaction, P < 0.05 acetate: propionate ratio for untreated straw. Greater relative populations of Ruminococcus albus (P < 0.05 and increased microbial N production (P = 0.045 were observed in Fast rumen inoculum. AFEX straw in Fast inoculum had greater total bacterial populations than Slow, but for untreated straw this result was reversed (interaction, P = 0.013. These findings indicate that differences in microbial populations in rumen fluid contribute to differences in the capacity of rumen inoculum to digest fiber.

Non-protein nitrogen and reduced phosphorus supply to sheep. 1. Phosphorus, volatile fatty acids, NH3 and pH of rumen liquid and daily fluid and phosphorus outflow from the rumen

International Nuclear Information System (INIS)

Hoeller, H.; Breves, G.; Martens, H.

1983-01-01

Sheep were fitted with cannulas in the rumen and in the abomasum and fed a straw/pellet diet providing 29% of total N as urea-N. The diet was deficient in P, and daily P intake during P depletion was about 1.3g. Phosphorus repletion was obtained in the same sheep by daily infusion of 2.85gP as phosphoric acid into the abomasum. During experimental periods, Cr-EDTA and 15 N-urea were continuously infused into the rumen for five days. During the last two days, blood samples and a total of ten samples from both rumen and abomasum were taken. Three depletion and three repletion experiments were performed. The following significant changes were noted in the P-deficient state (means): blood plasma inorganic P fell from 2.38 to 0.67mmol/L and rumen liquid total P from 5.65 to 1.71mmol/L, i.e. both levels decreased by approximately 70%. Fluid outflow from the rumen was raised by about 10% but P outflow decreased by about 68%. NH 3 concentrations in rumen fluid increased from 3.60 to 7.51mmol/L whereas total VFA concentration was reduced from 125 to 100mmol/L. Average pH values changed from 6.06 to 6.56. It is suggested that P depletion led to reduced microbial growth and activity in the rumen although data on microbial protein outflow from the rumen are not yet available. (author)

Influence of Albizia lebbeck Saponin and Its Fractions on In Vitro Gas Production Kinetics, Rumen Methanogenesis, and Rumen Fermentation Characteristics.

Science.gov (United States)

Sirohi, Sunil Kumar; Goel, Navneet; Singh, Nasib

2014-01-01

The present study was undertaken to investigate the effect of crude seed powder (CSP) and gross saponins extract (GSE) of seeds of Albizia lebbeck on antimicrobial activity by taking two Gram-positive (Staphylococcus aureus and Bacillus cereus), two Gram-negative (Escherichia coli and Salmonella Typhi) bacteria, and two fungi species (Aspergillus niger and Candida butyric) were taken at 25, 50, 100, 250, and 500 µg levels using agar well diffusion method. Zone of inhibition was increased with increasing of concentration of CSP and saponins which indicates that Gram-negative bacteria (E. coli), Gram-positive bacteria (B. cereus), and A. niger were significantly susceptible to inhibition. Another experiment was conducted to study the effect of GSE and saponins fraction A and B of A. lebbeck supplementation at 6% on DM basis on methane production and other rumen fermentation parameters using in vitro gas production test, by taking three different type diets, that is, high fiber diet (D1, 60R : 40C), medium fiber diet (D2, 50R : 50C), and low fiber diet (D3, 40R : 60C). Significant (P ≤ 0.05) increase was seen in IVDMD, methane production; however ammonia nitrogen concentration decreased as compared to control. The methane production was reduced in a range between 12 and 49% by saponin supplemented diets except in case of GSE in D2. Sap A showed the highest methane reduction per 200 mg of truly digested substrate (TDS) than other treatment groups. Results in relation with quantification of methanogens and protozoa by qPCR indicated the decreasing trend with saponins of A. lebbek in comparison with control except total methanogen quantified using mcr-A based primer.

Reaction of some rumen micro flora to different supplementary feeds ...

African Journals Online (AJOL)

Ruminant animals lack enzymes to break down fibrous feeds but they harbor microorganisms capable of degrading their feeds. Rumen microbes are affected by feed substrates. The purpose of this study was to evaluate rumen microbial changes as the function of varying supplementary feeds. Two protein supplements ...

The effect of pineapple waste (Ananas comosus (L. Merr subtitution on mixed basal diet of Elephant grass and calliandra on rumen ecosystem of sheep

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Y. Widiawati

2009-12-01

Full Text Available The aim of this study was to evaluate the effect of pineapple waste substitution to mixed basal diet of Elephant grass and calliandra on rumen ecosystem. Pineapple waste was substituted to basal of Elephant grass and calliandra leaves (3:2 at the level of 0% (RA; 10% (RB; 20% (RC; 30% (RD; 40% (RE and 50% (RF. In this experiment 24 Indonesian local male sheep (9-10 months old, 15.3 kg average body weight were used, and were divided into 6 groups of dietary treatment (4 sheep each. Every group was offered one of the experimental diets (RA to RF in a Completely Randomized Design. Pineapple waste was offered gradually for one month until the level of 50% (RF was reached. The animals were adapted to experimental diets for about 14 days prior to the data collection period. Rumen fluids from each animal was taken (5 hours after morning feeding for pH, ammonia concentration; bacteria and protozoa population analysis. The results showed that substitution of pineapple waste up to 30% had no effect on pH, but when the level was increased up to 40 and 50%, the pH (P0.05. Increasing pineapple waste given increased population of amyllolytic bacteria but decreased the population of cellulolytic bacteria. On the Elephant grass and calliandra basal diet with the proportion of 3 : 2, the best substitution of pineapple waste was up to 20%.

Effects of juniper essential oil on growth performance, some rumen protozoa, rumen fermentation and antioxidant blood enzyme parameters of growing Saanen kids.

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Yesilbag, D; Biricik, H; Cetin, I; Kara, C; Meral, Y; Cengiz, S S; Orman, A; Udum, D

2017-10-01

This study aimed to evaluate the effects of juniper essential oil on the growth performance, rumen fermentation parameters, rumen protozoa population, blood antioxidant enzyme parameters and faecal content in growing Saanen kids. Thirty-six male Saanen kids (36 ± 14 days of age) were used in the study. Each group consisted of 9 kids. The control group (G1) was fed with a diet that consisted of the above concentrated feed and oat hay, whereas the experimental groups consumed the same diet but with the concentrated feed uniformly sprayed with juniper essential oil 0.4 ml/kg (G2), 0.8 ml/kg (G3) or 2 ml/kg (G4). There were no differences (p > 0.05) in live weight, live weight gain or feed consumption between the control and experimental groups. There was a significant improvement (p rumen pH, rumen volatile fatty acid (VFA) profile or faecal pH of the control and experimental groups. The rumen NH 3 N values were similar at the middle and end of the experiment, but at the start of the experiment, the rumen NH 3 N values differed between the control and experimental groups (p < 0.05). The faecal score value was significantly (p < 0.05) decreased in the experimental groups. The addition of juniper essential oil supplementation to the rations caused significant effects on the kids' antioxidant blood parameters. Although the superoxide dismutase (SOD) activity, total antioxidant capacity (TAC) and catalase values were significantly (p < 0.05) increased in the experimental groups (G2, G3 and G4), especially group G4, the blood glutathione peroxidase (GPX) value significantly decreased in the experimental groups. The results of this study suggest that supplementation of juniper oil is more effective on antioxidant parameters than on performance parameters and may be used as a natural antioxidant product. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

Effect of sugar fatty acid esters on rumen fermentation in vitro

OpenAIRE

Wakita, M.; Hoshino, S.

1987-01-01

1.The effect of sugar fatty acid esters (SFEs; currently used as food additives for human consumption) on rumen volatile fatty acids (VFA) and gas production was studied with sheep rumen contents in vitro.2. Some SFEs having monoester contents of more than 70% increased the molar proportion of propionate in conjunction with reduction in the acetate: propionate ratio when the individual SFE was added to rumen contents in a final concentration of 4 g/l. Laurate sugar ester was the most potent p...

Rumen microbial changes in cattle fed diets with or without salinomycin.

OpenAIRE

Olumeyan, D B; Nagaraja, T G; Miller, G W; Frey, R A; Boyer, J E

1986-01-01

Four rumen-fistulated steers, randomly assigned to two groups (control and salinomycin fed) were used to monitor the changes in rumen microbial populations and volatile fatty acids (VFA) concentrations associated with feeding salinomycin (0.22 mg X kg-1 X day-1). Steers were adapted to an alfalfa hay and grain (80:20) diet before supplementing the diet with salinomycin, and then the diet was changed to 50:50 and 20:80 ratios of alfalfa hay to grain at 2-week intervals. Rumen samples for total...

Selection and production of bacteria which detoxify mimosine: Leucaena leaves may be used as ruminant feed

Energy Technology Data Exchange (ETDEWEB)

Boehnel, H. [Institute of Tropical Animal Health, Georg-August-University Goettingen (Germany)], E-mail: hboehne@gwdg.de; Aung, A [Institute of Tropical Animal Health, Georg-August-University Goettingen (Germany); University of Veterinary Sciences, Yezin (Myanmar)

2009-07-01

Mimosaceae are shrubs or trees, which grow abundantly in tropical regions. Their leaves contain high value protein which cannot be used as feed due to the toxic substance mimosine and its metabolites in the digestive tract These alkaloids cause diseases in ruminants, mainly loss of hair/wool, and may lead to death in higher concentration. This is the reason why the nutritive value cannot be exploited reasonably in animal production. Experience has shown that there are some geographical regions where animals do not suffer by mimosine. It was found that they harbour ruminal bacteria, which degrade mimosine to non-toxic metabolites. In cooperation with other microbes in the digestive tract, the full dietetic value of Leucaena may be exploited. To date several bacteria were isolated and may be used as feed additive, e.g., S. jonesii. However, their production and storage is expensive and complicated. A practical method is to orally inoculate animals and use their rumen fluid directly as feed additive. This rumen culture or cultured anaerobic bacteria in the laboratory may suffer during transport and storage; hence, they need a cold chain until the target animal. Our institute has an outstanding experience to produce bacterial veterinary vaccines, probiotics, and biological fertilizers in tropical countries. Continuous culture in a bioreactor is the base for the success. It was the idea to use this technology for selection and production of mimosine degrading bacteria. The presentation will give a short theoretical background of - bacterial fermenter production - continuous culture - isolation of specific bacteria under selective environmental conditions - mass production in a bioreactor to be used in tropical areas - purification and concentration of the bacterial crop by rinsing and hollow fibre cross flow ultra filtration - stabilisation of the product in alginate beads - microbiota in the digestive tract. The practical work started with ruminal content of German

[Isolation and identification of rumen bacteria for cellulolytic enzyme production].

Science.gov (United States)

Aihemaiti, Maierhaba; Zhen, Fan; Li, Yuezhong; Aibaidoula, Gulisimayi; Yimit, Wusiman

2013-05-04

We screened aerobic bacteria with cellulolytic activity from ruminal fluid of sheep, cattle and camel in Xinjiang. Fresh ruminal fluid was inoculated on sterilized sodium carboxymethylcellulose agar plates. Highly cellulolytic aerobic bacteria were screened out by using Congo red staining and liquid secondary screening culture media. The combination of morphological and biochemical test with 16SrDNA sequence analysis were used to classify the strains. Enzymatic activities of four strains with strong cellulose-decomposing abilities were studied under different culture conditions. Out 84 isolated cellulolytic strains, 40 exhibited strong abilities in decomposing cellulose. They are including 37 Gram-negative isolates and 3 Gram-positive strains. Identification of these 40 strains shows that they belong to 11 species of 6 genera, 16 strains in Stenotrophomonas maltophilia, 10 Ochrobactrum, 5 Sphingobacterium, 3 Microbacterium, 3 Paracoccus and 2 Pseudomonas. The results of the enzymatic studies of four strains with strong cellulolytic abilities indicates that the strains have the best enzyme producing property when straw powder was chosen as the carbon source; the pH at 5.5 -6.0 and temperature at 37 degrees C. The strains with highly cellulolytic abilities isolated from ruminal fluid show strong abilities in cellulose decomposition.

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

Science.gov (United States)

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; Prumen 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.

Development of Simple and Precise Method of Arginine Determination in Rumen Fluid by Spectrophotometer

International Nuclear Information System (INIS)

Chacher, B.; Marghazani, I. B.; Liu, J. X.; Liu, H. Y.

2015-01-01

The objective of current study was to build up a convenient, economic and accurate procedure to determine arginine (ARG) concentration in rumen fluid. Rumen fluid was collected from 3 rumen fistulated Chinese Holstein dairy cows and added with or without (control) 1mmol/l unprotected ARG and blank (with only medium) in to syringe system in triplicate as a replicate. All syringes were incubated in water bath at 39 Degree C for 0, 2, 4, 6, 12 and 24 h and were terminated to measure the ARG concentration. Sakaguchi reaction method was used to analyze the ARG concentration in rumen fluid by determining the rumen degradation rate of protected and unprotected ARG. Temperature, time and absorbance were optimized in the procedure based on Sakaguchi reaction. Color consistency remained 4-6 min. The optimum temperature (0-5) Degree C was observed for maximum optical density 0.663 at wave length 500 nm. Minimum ARG that could be determined in rumen fluid by spectrophotometer was 4-5 μ g/ml. No significance (P>0.05) difference were observed between two results derived from spectrophotometer and amino acid analyzer methods. In conclusion, the spectrophotometer method of ARG determination in rumen fluid based on Sakaguchi reaction is easy, accurate, and economical and could be useful in learning ARG metabolism in the rumen. (author)

Entry of blood urea into the rumen of the llama

International Nuclear Information System (INIS)

Hinderer, S.; Engelhardt, W. von

1976-01-01

Transfer of body urea into the temporarily isolated rumen cleaned and filled with a test solution was measured in the llama. Simultaneously urea recirculation into the total gastro-intestinal tract (GI-tract) was estimated using 14 C-urea. The permeability of the rumen wall to urea could be changed significantly. With CO 2 or butyric acid in the test solution, permeability was highest, it was low when food was withheld and when no volatile fatty acids were present in the solution. Changes in the permeability can affect the transfer of blood urea across the rumen wall more extensively than changes in plasma urea concentrations. (author)

The effects of plant extracts on microbial community structure in a rumen-simulating continuous-culture system as revealed by molecular profiling.

Science.gov (United States)

Ferme, D; Banjac, M; Calsamiglia, S; Busquet, M; Kamel, C; Avgustin, G

2004-01-01

An in vitro study in dual-flow continuous-culture fermentors was conducted with two different concentrations of monensin, cinnamaldehyde or garlic extract added to 1:1 forage-to-concentrate diet in order to determine their effects on selected rumen bacterial populations. Samples were subjected to total DNA extraction, restriction analysis of PCR amplified parts of 16S rRNA genes (ARDRA) and subsequent analysis of the restriction profiles by lab-on-chip technology with the Agilent's Bioanalyser 2100. Eub338-BacPre primer pair was used to select for the bacteria from the genera Bacteroides, Porphyromonas and Prevotella, especially the latter representing the dominant Gram-negative bacterial population in the rumen. Preliminary results of HaeIII restriction analysis show that the effects of monensin, cinnamaldehyde and garlic extract on the BacPre targeted ruminal bacteria are somewhat different in regard to targeted populations and to the nature of the effect. Garlic extract was found to trigger the most intensive changes in the structure of the BacPre targeted population. Comparison of the in silico restriction analysis of BacPre sequences deposited in different DNA databanks and of the results of performed amplified ribosomal DNA restriction analysis showed differences between the predicted and obtained HaeIII restriction profiles, and suggested the presence of novel, still unknown Prevotella populations in studied samples.

Use of Lysozyme as a Feed Additive on Rumen Fermentation and Methane Emission

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Ashraf A. Biswas

2016-11-01

Full Text Available This study was conducted to determine the effect of lysozyme addition on in vitro rumen fermentation and to identify the lysozyme inclusion rate for abating methane (CH4 production. An in vitro ruminal fermentation technique was done using a commercial concentrate to rice straw ratio of 8:2 as substrate. The following treatments were applied wherein lysozyme was added into 1 mg dry matter substrate at different levels of inclusion: Without lysozyme, 2,000, 4,000, and 8,000 U lysozyme. Results revealed that, lysozyme addition had a significant effect on pH after 24 h of incubation, with the highest pH (p<0.01 observed in 8,000 U lysozyme, followed by the 4,000 U, 2,000 U, and without lysozyme. The highest amounts of acetic acid, propionic acid (p<0.01 and total volatile fatty acid (TVFA (p<0.05 were found in 8,000 U after 24 h of incubation. The CH4 concentration was the lowest in the 8,000 U and the highest in the without lysozyme addition after 24 h of incubation. There was no significant differences in general bacteria, methanogen, or protozoan DNA copy number. So far, addition of lysozyme increased the acetate, propionate, TVFA, and decreased CH4 concentration. These results suggest that lysozyme supplementation may improve in vitro rumen fermentation and reduce CH4 emission.

Development of Equation Based on Urinary Purine Derivatives to Estimate Rumen Microbial Protein Production in Goats

International Nuclear Information System (INIS)

Jetana, Thongsuk; Abdullah, Norhani; Liang, Boo Juan; Syed Salim, Syed Jalaludin; Ho, Wan Yin

2003-06-01

Three experiments were conducted at the farm of the Universiti Putra Malaysia, Serdang, Selangor, Malaysia, to establish a model as an index for estimating rumen microbial protein production. In Experiment 1, six Ferral male goats (wt. 40.2±4.6 kg) were used to determine the endogenous purine derivatives (PD) excreted in the urine by fasting. In Experiment 2, four Ferral male goats (wt. 39.6±1.8 kg) were used to measure the proportion of plasma PD excreted in the urine by using [ 14 C]-uric acid as a marker at two levels of feed intake (40% and 80% voluntary intake), using an incomplete 2x4 Latin square experimental design. The feed consisted of 40% oil palm frond and 60% concentrate (OPFC). In Experiment 3, four Ferral male goats fed (OPFC)) were slaughtered and rumen contents were taken for measurements of purine and total nitrogen contents of mixed rumen microbes. The results showed that endogenous PD (allantoin, uric acid, xanthine and hypoxanthine) excreted in the urine obtained by the fasting trial was 202±17 μmol/kg BW 0 . 75 d - 1. The average percentage recovery of plasma PD excretion in the urine by using [ 14 C)-uric acid as a marker was 83±2.0% (cv=6.88, ranged 76.3-91.4%, n=8). Percentage recovery was not affected by levels of feed intake. The ratio of purine N: total N in the mixed rumen liquid associated bacteria (LAB) was 0.085. In this study, a preliminary model for goats was established by using the information from the recovery of labeled PD [ 14 C]-uric acid and the fasting PD excretion. The model obtained was Y 0.83X + 0.202 x BW 0 . 75 , where Y = PD excretion in the urine (mmol/d) X PD absorption at small intestine (mmol/d) BW 0 . 75 = Metabolic body weight (kg) Thus the microbial nitrogen based on total PD (MNpd) can be calculated as follows: MNpd = 70 x X = 0.992 x X (g/d) 0.085 x 0.83 x 1000 where 0.085 is the ratio of purine-N: total N in mixed rumen microbes, 0.83 is the average of digestibility of microbial purine from published

Some aspects of nitrogen metabolism in the bovine rumen

International Nuclear Information System (INIS)

Nikolic, J.A.

1976-01-01

Studies on the use of urea as a source of nitrogen for microbial protein synthesis showed that utilization was more efficient when the protein-N content of the diet was low. However, when the total nitrogen of the dietary dry matter was reduced below 1.9%, there was a drop in the protein-N content of rumen dry matter and a reduction in the daily flow of protein through the duodenum. The mean post-prandial rumen ammonia concentrations were below 5mg NH 3 -N/100ml. In vitro work with 15 N-labelled urea and ammonia salts showed that the overall utilization rate of ammonia was not significantly affected by mean concentrations between 1.6 and 16.7mg/ml, but that net utilization was lower below 5.8mg/100ml. It is suggested that increased lysis of susceptible micro-organisms or increased proteolysis of feed proteins may account for these findings. The concentrations of ammonia-N, protein-N and volatile fatty acids in the rumen were not affected by increases in the amount of calcium, phosphorus, potassium or sulphur in the diet, although the concentrations of these elements were significantly increased in the rumen. A wide dietary Ca/P ratio (3.26) tended to reduce the stability of rumen contents leading to a low pH and foaming. Increases in mean sulphide concentration from 3.6 to 8.0mg/l had no effect on overall protein synthesis rate as indicated by the 35 S incorporation rate. (author)

Dose-response effects of dietary pequi oil on fermentation characteristics and microbial population using a rumen simulation technique (Rusitec).

Science.gov (United States)

Duarte, Andrea Camacho; Durmic, Zoey; Vercoe, Philip E; Chaves, Alexandre V

2017-12-01

The effect of increasing the concentration of commercial pequi (Caryocar brasiliense) oil on fermentation characteristics and abundance of methanogens and fibrolityc bacteria was evaluated using the rumen simulation technique (Rusitec). In vitro incubation was performed over 15 days using a basal diet consisting of ryegrass, maize silage and concentrate in equal proportions. Treatments consisted of control diet (no pequi oil inclusion, 0 g/kg DM), pequi dose 1 (45 g/kg DM), and pequi dose 2 (91 g/kg DM). After a 7 day adaptation period, samples for fermentation parameters (total gas, methane, and VFA production) were taken on a daily basis. Quantitative real time PCR (q-PCR) was used to evaluate the abundance of the main rumen cellulolytic bacteria, as well as abundance of methanogens. Supplementation with pequi oil did not reduce overall methane production (P = 0.97), however a tendency (P = 0.06) to decrease proportion of methane in overall microbial gas was observed. Increasing addition of pequi oil was associated with a linear decrease (P < 0.01) in dry matter disappearance of maize silage. The abundance of total methanogens was unchanged by the addition of pequi oil, but numbers of those belonging to Methanomassiliicoccaceae decreased in liquid-associated microbes (LAM) samples (P < 0.01) and solid-associated microbes (SAM) samples (P = 0.09) respectively, while Methanobrevibacter spp. increased (P < 0.01) only in SAM samples. Fibrobacter succinogenes decreased (P < 0.01) in both LAM and SAM samples when substrates were supplemented with pequi oil. In conclusion, pequi oil was ineffective in mitigating methane emissions and had some adverse effects on digestibility and selected fibrolytic bacteria. Copyright © 2017 Elsevier Ltd. All rights reserved.

Isolation and Identification of Bacteria That Has Potential as Producer of Protease Enzyme in the Tannery Industry, PT. Adi Satria Abadi (ASA), YOGYAKARTA

OpenAIRE

Said, M. I; Likadja, J. C

2012-01-01

Bacteria are one of the microorganisms that have the potential as a producer of protease enzyme. Tannery industrial waste is one of the media predicted to contain a number of proteolytic bacteria because of the waste generated is composed largely of protein and fat which are good as growing medium for bacteria. This study aimed to isolate and identify bacteria that have the potential as a producer of protease enzyme. Research conducted at the waste water processing installation (WWPI), tanner...

Use of Proteolytic Enzymes in the Treatment of Proteinaceous Esophageal Food Impaction.

Science.gov (United States)

Morse, Christopher R; Wang, Howard; Donahue, Dean M; Garrity, Julie M; Allan, James S

2016-01-01

Proteinaceous esophageal food impaction typically requires endoscopic intervention. An alternative approach is the use of proteolytic enzymes. Concerns regarding the use of proteolytic enzymes include the risk of perforation and aspiration pneumonitis. We retrospectively reviewed our series of 69 patients treated with papain to determine the safety and efficacy of proteolytic enzymes. Patients were retrospectively reviewed if treated for an esophageal food impaction from 1999 through 2008. Median age was 56 years (range 19-91 years), with 46 male and 23 female patients. In 27 patients (39%) this was their first presentation, in 14 (20%) it was the second, and 28 (41%) had multiple previous episodes. Meat was the cause in 49 (71%), chicken in 6 (9%), fish in 3 (4%), and unspecified in 11 (16%). All patients presented with dysphagia for solids, 56 (81%) could not tolerate liquids. Papain solution, 1 tsp in 8 oz of water, was given to patients in an unlimited quantity. Papain was successful in relieving the obstruction in 60 patients (87%). The remaining 9 patients (13%) underwent endoscopy with successful retrieval. No patient suffered a perforation, either with papain ingestion or endoscopy. There were no episodes of pneumonitis or pneumonia. We have used proteolytic enzymes with a high success rate and with minimal complication. Further, if proteolytic enzymes fail, endoscopy can be performed safely and effectively. We recommend the use of proteolytic enzymes as the initial management in all patients with proteinaceous food impaction of the esophagus. Published by Elsevier Inc.

A Structural and Functional Elucidation of the Rumen Microbiome Influenced by Various Diets and Microenvironments

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Simon Deusch

2017-08-01

Full Text Available The structure and function of the microbiome inhabiting the rumen are, amongst other factors, mainly shaped by the animal's feed intake. Describing the influence of different diets on the inherent community arrangement and associated metabolic activities of the most active ruminal fractions (bacteria and archaea is of great interest for animal nutrition, biotechnology, and climatology. Samples were obtained from three fistulated Jersey cows rotationally fed with corn silage, grass silage or grass hay, each supplemented with a concentrate mixture. Samples were fractionated into ruminal fluid, particle-associated rumen liquid, and solid matter. DNA, proteins and metabolites were analyzed subsequently. DNA extracts were used for Illumina sequencing of the 16S rRNA gene and the metabolomes of rumen fluids were determined by 500 MHz-NMR spectroscopy. Tryptic peptides derived from protein extracts were measured by LC-ESI-MS/MS and spectra were processed by a two-step database search for quantitative metaproteome characterization. Data are available via ProteomeXchange with the identifier PXD006070. Protein- and DNA-based datasets revealed significant differences between sample fractions and diets and affirmed similar trends concerning shifts in phylogenetic composition. Ribosomal genes and proteins belonging to the phylum of Proteobacteria, particularly Succinivibrionaceae, exhibited a higher abundance in corn silage-based samples while fiber-degraders of the Lachnospiraceae family emerged in great quantities throughout the solid phase fractions. The analysis of 8163 quantified bacterial proteins revealed the presence of 166 carbohydrate active enzymes in varying abundance. Cellulosome affiliated proteins were less expressed in the grass silage, glycoside hydrolases appeared in slightest numbers in the corn silage. Most expressed glycoside hydrolases belonged to families 57 and 2. Enzymes analogous to ABC transporters for amino acids and

Proteolytic activity regarding Sarconesiopsis magellanica (Diptera: Calliphoridae) larval excretions and secretions.

Science.gov (United States)

Pinilla, Yudi T; Moreno-Pérez, Darwin A; Patarroyo, Manuel A; Bello, Felio J

2013-12-01

Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Diptera maggots release proteolytic enzymes contained in larval excretion and secretion (ES) products playing a key role in digestion. Special interest in proteolytic enzymes has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue during larval therapy. This study was thus aimed at identifying and characterising S. magellanica proteolytic enzyme ES products for the first time. These products were obtained from first-, second- and third-instar larvae taken from a previously-established colony. ES proteins were separated by SDS-PAGE and their proteolytic activity was characterised by zymograms and inhibition assays involving BAPNA (Nα-benzoyl-dl-Arg-p-nitroanilide) and SAPNA substrates, using synthetic inhibitors. The protein profile ranged from ∼69kDa to ∼23kDa; several of them coincided with the Lucilia sericata ES protein profile. Serine-protease hydrolysis activity (measured by zymogram) was confirmed when a ∼25kDa band disappeared upon ES incubation with PMSF inhibitor at pH 7.8. Analysis of larval ES proteolytic activity on BAPNA and SAPNA substrates (determined by using TLCK and TPCK specific inhibitors) suggested a greater amount of trypsin-like protease. These results support the need for further experiments aimed at validating S. magellanica use in larval therapy. Copyright © 2013 Elsevier B.V. All rights reserved.

The Effect of DNA Extraction Methods on Observed Microbial Communities from Fibrous and Liquid Rumen Fractions of Dairy Cows

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Jueeli D. Vaidya

2018-01-01

summary, the comprehensive assessment of observed communities of bacteria, archaea and anaerobic fungi described here provides insight into a rational basis for selecting an optimal methodology to obtain a representative picture of the rumen microbiota.

Purification Techniques of Bacteriocins from Lactic Acid Bacteria and Other Gram-Positive Bacteria

Science.gov (United States)

Saavedra, Lucila; Sesma, Fernando

The search for new antimicrobial peptides produced by lactic acid ­bacteria and other Gram-positive microorganisms has become an interesting field of research in the past decades. The fact that bacteriocins are active against numerous foodborne and human pathogens, are produced by generally regarded as safe (GRAS) microorganisms, and are readily degraded by proteolytic host systems makes them attractive candidates for biotechnological applications. However, before suggesting or choosing a new bacteriocin for future technology developments, it is necessary to elucidate its biochemical structure and its mode of action, which may be carried out once the bacteriocin is purified to homogeneity. This chapter focuses on describing the main strategies used for the purification of numerous bacteriocins.

Histamine Induces Bovine Rumen Epithelial Cell Inflammatory Response via NF-κB Pathway

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Xudong Sun

2017-06-01

Full Text Available Background/Aims: Subacute ruminal acidosis (SARA is a common disease in high-producing lactating cows. Rumenitis is the initial insult of SARA and is associated with the high concentrations of histamine produced in the rumen of dairy cows during SARA. However, the exact mechanism remains unclear. The objective of the current study is to investigate whether histamine induces inflammation of rumen epithelial cells and the underlying mechanism of this process. Methods: Bovine rumen epithelial cells were cultured and treated with different concentrations of histamine and pyrrolidine dithiocarbamate (PDTC, an NF-κB inhibitor cultured in different pH medium (pH 7.2 or 5.5. qRT-PCR, Western-blotting, ELISA and immunocytofluorescence were used to evaluate whether histamine activated the NF-κB pathway and inflammatory cytokines. Results: The results showed that histamine significantly increased the activity of IKK β and the phosphorylation levels of IκB α, as well as upregulated the mRNA and protein expression levels of NF-κB p65 in the rumen epithelial cells cultured in neutral (pH=7.2 and acidic (pH=5.5 medium. Furthermore, histamine treatment also significantly increased the transcriptional activity of NF-κB p65. High expression and transcriptional activity of NF-κB p65 significantly increased the mRNA expressions and concentrations of inflammatory cytokines, tumor necrosis factor alpha (TNF-α, interleukin 6 (IL-6 and interleukin 1 beta (IL-1β, thereby inducing the inflammatory response in bovine rumen epithelial cells. However, inhibition of NF-κB p65 by PDTC significantly decreased the expressions and concentrations of the inflammatory cytokines induced by histamine in the rumen epithelial cells cultured in the neutral and acidic medium. Conclusion: The present data indicate that histamine induces the inflammatory response of bovine rumen epithelial cells through the NF-κB pathway.

Degradation of lucerne stem cell walls by five rumen bacterial species

NARCIS (Netherlands)

Jung, H.G.; Engels, F.M.; Weimer, P.J.

2004-01-01

The rumen bacterial strains Butyrivibrio fibrisolvens H17c, Fibrobacter succinogenes S85, Lachnospira multiparus 40, Ruminococcus albus 7 and R. flavefaciens FD-1 were compared individually and as a five-species mixture with a rumen inoculum for their ability to degrade lucerne (Medicago sativa L.)

Relations between passage rates of rumen fluid and particulate matter and foam production in rumen contents of cattle fed on different diets ad lib.

Science.gov (United States)

Okine, E K; Mathison, G W; Hardin, R T

1989-03-01

1. A group of six cattle, three of which had a non-bloating history (group A) and had been ruminally cannulated for the previous 2 years, and three with a history of being bloat-prone (group B) and which had been ruminally cannulated only 3 months before the study, were fed ad lib. on chopped lucerne (Medicago sativa) hay, lucerne pellets, or a 100 g chopped hay and 900 g rolled barley grain/kg diet over three periods of 30 d each. Flow of rumen digesta, by reference to CoEDTA and chromium-mordanted fibres, and foam production from samples of rumen contents were measured. 2. Samples of rumen contents (50 ml) from group A produced foam heights of 150 and 60 mm, 2 and 4 h after feeding respectively, compared with 240 and 150 mm for group B (P less than 0.05). 3. The fractional passage rate of the 1-2 mm particles mordanted with Cr did not differ (P greater than 0.05) between groups. 4. The fractional outflow rates (FOR) for CoEDTA 0-2 h and 2-7 h after feed was offered were 0.205 and 0.160/h for group A and 0.093 and 0.086/h for group B respectively (P less than 0.05). 5. Rumen-fluid FOR 0-2 h and 2-7 h after provision of feed were significantly (P less than 0.05) inversely correlated (r -0.74 and -0.85 respectively) with the amount of foam produced from rumen contents at these times.

Isolation and removal of proteolytic enzymes with magnetic cross-linked erythrocytes

International Nuclear Information System (INIS)

Safarik, I. Ivo; Safarikova, Mirka

2001-01-01

New magnetic adsorbents for batch isolation and removal of various proteolytic enzymes were prepared by glutaraldehyde cross-linking of bovine, porcine and human erythrocytes in the presence of fine magnetic particles. Trypsin, chymotrypsin, alkaline bacterial protease and proteases present in various commercial enzyme preparations were efficiently adsorbed on these adsorbents; on the contrary, proteins without proteolytic activity were not adsorbed

A note on the insertion of rumen cannulae in pregnant ewes

African Journals Online (AJOL)

Modifications to a simple and rapid technique for the insertion of rumen cannulae in sheep are described. The modified technique, executed in three phases, was developed to facilitate the fistula- tion and insertion of rumen cannulae in pregnant ewes, especially during late pregnancy. Wysigings van 'n eenvoudige en ...

A Study on Rumen Cilliate Protozoa Population, pH and some ...

African Journals Online (AJOL)

An investigation on rumen ciliate protozoa population, pH and some metabolites (total volatile fatty acids, rumen ammonia Nitrogen) was conducted on two fistulated WAD rams fed forage and concentrate diets. The 12-week study focused on the sequence of production of these parameters under each dietary regime.

Nutrient intake, digestibility and rumen metabolites in bulls fed rice ...

African Journals Online (AJOL)

Nutrient intake, digestibility and rumen metabolites were determined in rumen - cannulated bulls fed rice straw or straw supplemented with urea, groundnut hay or cotton seed cake. Total dry matter intake (DMI) ranged from 7.55 Lo 8.29kg/d or 3.66 to 4.04% of liveweight and from 6.48 to 7. 21 kg/d for organic matter.

Role of Proteolytic Enzymes in the Interaction of Phytopathogenic Microorganisms with Plants.

Science.gov (United States)

Valueva, T A; Zaichik, B Ts; Kudryavtseva, N N

2016-12-01

Various forms of participation of proteolytic enzymes in pathogenesis and defense in plants are reviewed. Along with extracellular proteinases, phytopathogenic microorganisms produce specific effectors having proteolytic activity and capable of acting on proteins inside plant cells. In turn, for defense against pathogens, plants use both extracellular and intracellular proteinases.

Efficacy of different essential oils in modulating rumen fermentation in vitro using buffalo rumen liquor

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Debashis Roy

2014-04-01

Full Text Available Aim: Present study was conducted to examine the modulatory effect of different essential oils on rumen fermentation pattern in vitro using wheat straw based diet (concentrate: wheat straw 50:50. Materials and Methods: Four essential oils i.e. cinnamon, garlic, oregano and rosemary oils were tested at concentration of 0, 30, 300 and 600 mg/litre (ppm of total culture fluid using in vitro gas production technique. Total gas production, methane production, nutrient degradability, volatile fatty acid (VFA production and ammonia nitrogen concentration were studied in vitro using buffalo rumen liquor. Results: Results indicated that all four essential oils decreased gas production significantly (P<0.05 at 600ppm concentration. However, in case of garlic oil, 300 ppm concentration was also found to be effective in decreasing total gas production. Reduction in methane production was found maximum (P<0.05 at higher doses in most of the oils. Maximum reduction in methane was noticed with garlic oil at 600ppm dose. Ammonia-N concentration was also decreased significantly (P<0.05 with essential oils and was found minimum with oregano oil at 600 ppm dose. Partition factor was found to be significantly (P<0.05 higher in 600 ppm concentration of garlic and oregano oil. The degradability of dry matter decreased significantly with higher concentration of essential oil in most of treatment combinations. Conclusion: Supplementation with different essential oils on wheat straw based diet modulates rumen fermentation and reduced methane and ammonia- N production and improved utilization of nutrients.

Detection of proteolytic signatures for Parkinson's disease

DEFF Research Database (Denmark)

Jordal, Peter Lüttge; Dyrlund, Thomas F.; Winge, Kristian

2016-01-01

Aim: To investigate if idiopathic Parkinson's disease (IPD) is associated with distinct proteolytic signatures relative to non-neurodegenerative controls (NND) and patients with multiple system atrophy (MSA). Materials & methods: A subtiligase-based N-terminomics screening method was exploited...

Biological and Proteolytic Variation in the Venom of Crotalus scutulatus scutulatus from Mexico

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Miguel Borja

2018-01-01

Full Text Available Rattlesnake venoms may be classified according to the presence/absence and relative abundance of the neurotoxic phospholipases A 2 s (PLA 2 s, such as Mojave toxin, and snake venom metalloproteinases (SVMPs. In Mexico, studies to determine venom variation in Mojave Rattlesnakes (Crotalus scutulatus scutulatus are limited and little is known about the biological and proteolytic activities in this species. Tissue (34 and venom (29 samples were obtained from C. s. scutulatus from different locations within their distribution in Mexico. Mojave toxin detection was carried out at the genomic (by PCR and protein (by ELISA levels for all tissue and venom samples. Biological activity was tested on representative venoms by measuring LD 50 and hemorrhagic activity. To determine the approximate amount of SVMPs, 15 venoms were separated by RP-HPLC and variation in protein profile and proteolytic activity was evaluated by SDS-PAGE (n = 28 and Hide Powder Azure proteolytic analysis (n = 27. Three types of venom were identified in Mexico which is comparable to the intraspecific venom diversity observed in the Sonoran Desert of Arizona, USA: Venom Type A (∼Type II, with Mojave toxin, highly toxic, lacking hemorrhagic activity, and with scarce proteolytic activity; Type B (∼Type I, without Mojave toxin, less toxic than Type A, highly hemorrhagic and proteolytic; and Type A + B, containing Mojave toxin, as toxic as venom Type A, variable in hemorrhagic activity and with intermediate proteolytic activity. We also detected a positive correlation between SVMP abundance and hemorrhagic and proteolytic activities. Although more sampling is necessary, our results suggest that venoms containing Mojave toxin and venom lacking this toxin are distributed in the northwest and southeast portions of the distribution in Mexico, respectively, while an intergradation in the middle of both zones is present.

Cellulosomics, a gene-centric approach to investigating the intraspecific diversity and adaptation of Ruminococcus flavefaciens within the rumen.

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Jennifer M Brulc

Full Text Available BACKGROUND: The bovine rumen maintains a diverse microbial community that serves to break down indigestible plant substrates. However, those bacteria specifically adapted to degrade cellulose, the major structural component of plant biomass, represent a fraction of the rumen microbiome. Previously, we proposed scaC as a candidate for phylotyping Ruminococcus flavefaciens, one of three major cellulolytic bacterial species isolated from the rumen. In the present report we examine the dynamics and diversity of scaC-types both within and between cattle temporally, following a dietary switch from corn-silage to grass-legume hay. These results were placed in the context of the overall bacterial population dynamics measured using the 16S rRNA. PRINCIPAL FINDINGS: As many as 117 scaC-types were estimated, although just nineteen were detected in each of three rumens tested, and these collectively accounted for the majority of all types present. Variation in scaC populations was observed between cattle, between planktonic and fiber-associated fractions and temporally over the six-week survey, and appeared related to scaC phylogeny. However, by the sixth week no significant separation of scaC populations was seen between animals, suggesting enrichment of a constrained set of scaC-types. Comparing the amino-acid translation of each scaC-type revealed sequence variation within part of the predicted dockerin module but strong conservation in the N-terminus, where the cohesin module is located. CONCLUSIONS: The R. flavefaciens species comprises a multiplicity of scaC-types in-vivo. Enrichment of particular scaC-types temporally, following a dietary switch, and between fractions along with the phylogenetic congruence suggests that functional differences exist between types. Observed differences in dockerin modules suggest at least part of the functional heterogeneity may be conferred by scaC. The polymorphic nature of scaC enables the relative distribution of R

QUANTIFICATION OF THE EFFICIENCY OF RUMEN MICROBIAL PROTEIN SYNTHESIS IN STEERS FED GREEN TROPICAL GRASS

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MARTHEN L. MULLIK

2012-09-01

Full Text Available ABSTRACT The rate of rumen microbial crude protein (MCP supply to the intestines is a crucial element in the current rumen models to predict respond of ruminants to a certain diet. Data from tropical pastures always below predicted results from the existing rumen models. Thus, quantification of the rumen MCP supply from tropical grass will improve predictive rate under tropical feeding conditions. Four Brahman crossbred steers (457±20.1 kg were used in a metabolism study. Pangola grass (Digitaria erianthe cv. Steudal was harvested every morning and fed to the animals soon after. Parameters measured were EMPS, intake, fractional passage rates, and rumen ammonia concentration. The EMPS was estimated using purine derivative excretion in urine. Crude protein and water soluble carbohydrates content were 6.3 and 7.4% of dry matter (DM respectively. DM intake was 1.6% live weight. Average rumen ammonia concentration was 69 mg/L whilst rumen passage rates were 7.84 and 6.92 %/h for fluid and solids respectively. EMPS was only 72 g MCP/kg digestible organic matter. It might be concluded that EMPS in steers consuming green pangola grass was below the minimum level for forage diets adopted in the current feeding standards. ABSTRAK Tingkat pasokan protein mikroba rumen (MCP ke usus halus merupakan salah satu unsur kunci dalam meramal respon pertumbuhan ruminan terhadap ransum tertentu. Data MCP hijauan tropis selalu berada di bawah nilai prediksi model rumen yang dipakai saat ini. Dengan demikian, kuantifikasi pasokan MCP rumput tropis diharapkan menjadi masukan untuk meningkatkan kemampuan prediksi model rumen untuk pakan daerah tropis. Empat sapi jantan muda Brahman persilangan (457±20,1 kg digunakan dalam sebuah penelitian metabolisme. Rumput pangola (Digitaria erianthe cv. Steudal dipanen setiap pagi dan langsung diberikan kepada ternak dalam kandang metabolis. Parameter yang diukur adalah produksi MCP dan efisiensi sintesis MCP (Emps, konsumsi, laju

Fatty acid oxidation products ('green odour') released from perennial ryegrass following biotic and abiotic stress, potentially have antimicrobial properties against the rumen microbiota resulting in decreased biohydrogenation.

Science.gov (United States)

Huws, S A; Scott, M B; Tweed, J K S; Lee, M R F

2013-11-01

In this experiment, we investigated the effect of 'green odour' products typical of those released from fresh forage postabiotic and biotic stresses on the rumen microbiota and lipid metabolism. Hydroperoxyoctadecatrienoic acid (HP), a combination of salicylic and jasmonic acid (T), and a combination of both (HPT) were incubated in vitro in the presence of freeze-dried ground silage and rumen fluid, under rumen-like conditions. 16S rRNA (16S cDNA) HaeIII-based terminal restriction fragment length polymorphism-based (T-RFLP) dendrograms, canonical analysis of principal coordinates graphs, peak number and Shanon-Weiner diversity indices show that HP, T and HPT likely had antimicrobial effects on the microbiota compared to control incubations. Following 6 h of in vitro incubation, 15.3% of 18:3n-3 and 4.4% of 18:2n-6 was biohydrogenated in control incubations, compared with 1.3, 9.4 and 8.3% of 18:3n-3 for HP, T and HPT treatments, respectively, with negligible 18:2n-6 biohydrogenation seen. T-RFLP peaks lost due to application of HP, T and HPT likely belonged to as yet uncultured bacteria within numerous genera. Hydroperoxyoctadecatrienoic acid, T and HPT released due to plant stress potentially have an antimicrobial effect on the rumen microbiota, which may explain the decreased biohydrogenation in vitro. These data suggest that these volatile chemicals may be responsible for the higher summer n-3 content of bovine milk. © 2013 The Society for Applied Microbiology.

Rumen morphometrics and the effect of digesta pH and volume on volatile fatty acid absorption.

Science.gov (United States)

Melo, L Q; Costa, S F; Lopes, F; Guerreiro, M C; Armentano, L E; Pereira, M N

2013-04-01

The effects of rumen digesta volume and pH on VFA absorption and its relation to rumen wall morphology were evaluated. Nine rumen cannulated cows formed 3 groups based on desired variation in rumen morphology: The High group was formed by Holsteins yielding 25.9 kg milk/d and fed on a high-grain total mixed ration (TMR); the Medium group by Holstein-Zebu crossbreds yielding 12.3 kg milk/d and fed on corn silage, tropical pasture, and a commercial concentrate; and the Dry group by nonlactating grazing Jerseys fed exclusively on tropical pasture. Within each group, a sequence of 3 ruminal conditions was induced on each cow in 3 × 3 Latin Squares, with 7-d periods: high digesta volume and high pH (HVHP), low volume and high pH (LVHP), and low volume and low pH (LVLP). Rumen mucosa was biopsied on the first day of Period 1. Ruminal morphometric variables evaluated were mitotic index, absorptive surface and papillae number per square centimeter of wall, area per papillae, papillae area as a percentage of absorptive surface, and epithelium, keratinized layer, and nonkeratinized layer thickness. There was marked variation in rumen morphology among the groups of cows. Grazing Jerseys had decreased rumen wall absorptive surface area and basal cells mitotic index, and increased thickness of the epithelium and of the keratin layer compared with cows receiving concentrates. Mean rumen pH throughout the 4 h sampling period was: 6.78 for HVHP, 7.08 for LVHP, and 5.90 for LVLP (P rumen wall to absorb VFA was estimated by the Valerate/CrEDTA technique. The fractional exponential decay rate for the ratio of valeric acid to Cr (k Val/Cr) was determined by rumen digesta sampling at 20-min intervals during 4 h, after the mixing of markers and the return of the evacuated ruminal content. The k Val/Cr values for treatments HVHP, LVHP, and LVLP were, respectively: 19.6, 23.9, and 35.0 %/h (SEM = 2.01; P = 0.21 for contrast HVHP vs. LVHP and P rumen wall and the mean of the 3 k Val

Responses of anaerobic rumen fungal diversity (phylum Neocallimastigomycota) to changes in bovine diet.

Science.gov (United States)

Boots, B; Lillis, L; Clipson, N; Petrie, K; Kenny, D A; Boland, T M; Doyle, E

2013-03-01

Anaerobic rumen fungi (Neocallimastigales) play important roles in the breakdown of complex, cellulose-rich material. Subsequent decomposition products are utilized by other microbes, including methanogens. The aim of this study was to determine the effects of dietary changes on anaerobic rumen fungi diversity. Altered diets through increasing concentrate/forage (50 : 50 vs 90 : 10) ratios and/or the addition of 6% soya oil were offered to steers and the Neocallimastigales community was assessed by PCR-based fingerprinting with specific primers within the barcode region. Both a decrease in fibre content and the addition of 6% soya oil affected Neocallimastigales diversity within solid and liquid rumen phases. The addition of 6% soya oil decreased species richness. Assemblages were strongly affected by the addition of 6% soya oil, whereas unexpectedly, the fibre decrease had less effect. Differences in volatile fatty acid contents (acetate, propionate and butyrate) were significantly associated with changes in Neocallimastigales assemblages between the treatments. Diet clearly influences Neocallimastigales assemblages. The data are interpreted in terms of interactions with other microbial groups involved in fermentation processes within the rumen. Knowledge on the influence of diet on anaerobic fungi is necessary to understand changes in microbial processes occurring within the rumen as this may impact on other rumen processes such as methane production. © 2012 The Society for Applied Microbiology.

Effects of isobutyrate supplementation on ruminal microflora, rumen enzyme activities and methane emissions in Simmental steers.

Science.gov (United States)

Wang, C; Liu, Q; Zhang, Y L; Pei, C X; Zhang, S L; Wang, Y X; Yang, W Z; Bai, Y S; Shi, Z G; Liu, X N

2015-02-01

The objective of this study was to evaluate the effects of isobutyrate supplementation on rumen microflora, enzyme activities and methane emissions in Simmental steers consuming a corn stover-based diet. Eight ruminally cannulated Simmental steers were used in a replicated 4 × 4 Latin square experiment. The treatments were control (without isobutyrate), low isobutyrate (LIB), moderate isobutyrate (MIB) and high isobutyrate (HIB) with 8.4, 16.8 and 25.2 g isobutyrate per steer per day respectively. Isobutyrate was hand-mixed into the concentrate portion. Diet consisted of 60% corn stover and 40% concentrate [dry matter (DM) basis]. Dry matter intake (averaged 9 kg/day) was restricted to a maximum of 90% of ad libitum intake. Population of total bacteria, cellulolytic bacteria and anaerobic fungi were linearly increased, whereas that of protozoa and total methanogens was linearly reduced with increasing isobutyrate supplementation. Real-time PCR quantification of population of Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio fibrisolvens and Fibrobacter succinogenes was linearly increased with increasing isobutyrate supplementation. Activities of carboxymethyl cellulase, xylanase and β-glucosidase were linearly increased, whereas that of protease was linearly reduced. Methane production was linearly decreased with increasing isobutyrate supplementation. Effective degradabilities of cellulose and hemicellulose of corn stover were linearly increased, whereas that of crude protein in diet was linearly decreased with increasing isobutyrate supplementation. The present results indicate that isobutyrate supplemented improved microflora, rumen enzyme activities and methane emissions in steers. It was suggested that the isobutyrate stimulated the digestive micro-organisms or enzymes in a dose-dependent manner. In the experimental conditions of this trial, the optimum isobutyrate dose was approximately 16.8 g isobutyrate per steer per day. Journal of Animal

Rumen protozoa and methanogenesis: not a simple cause-effect relationship.

Science.gov (United States)

Morgavi, Diego P; Martin, Cécile; Jouany, Jean-Pierre; Ranilla, Maria José

2012-02-01

Understanding the interactions between hydrogen producers and consumers in the rumen ecosystem is important for ruminant production and methane mitigation. The present study explored the relationships between rumen protozoa, methanogens and fermentation characteristics. A total of six donor sheep harbouring (F, faunated) or not (D, defaunated) protozoa in their rumens (D animals were kept without protozoa for a period of a few months (D - ) or for more than 2 years (D+)) were used in in vitro and in vivo experiments. In vitro the absence of protozoa decreased NH3 and butyrate production and had no effect on methane. In contrast, the liquid-associated bacterial and methanogens fraction of D+ inocula produced more methane than D -  and F inoculum (P protozoa may affect differently the methanogen community and methane emissions in wethers.

Continuous volatile fatty acid production from lignocellulosic biomass by a novel rumen-mimetic bioprocess.

Science.gov (United States)

Agematu, Hitosi; Takahashi, Takehiko; Hamano, Yoshio

2017-11-01

Lignocellulosic biomass is an attractive source of biofuels and biochemicals, being abundant in various plant sources. However, processing this type of biomass requires hydrolysis of cellulose. The proposed rumen-mimetic bioprocess consists of dry-pulverization of lignocellulosic biomass and pH-controlled continuous cultivation of ruminal bacteria using ammonium as a nitrogen source. In this study, ruminal bacteria were continuously cultivated for over 60 days and used to digest microcrystalline cellulose, rice straw, and Japanese cedar to produce volatile fatty acids (VFAs). The ruminal bacteria grew well in the chemically defined medium. The amounts of VFAs produced from 20 g of cellulose, rice straw, and Japanese cedar were 183 ± 29.7, 69.6 ± 12.2, and 21.8 ± 12.9 mmol, respectively. Each digestion completed within 24 h. The carbon yield was 60.6% when 180 mmol of VFAs was produced from 20 g of cellulose. During the cultivation, the bacteria were observed to form flocs that enfolded the feed particles. These flocs likely contain all of the bacterial species necessary to convert lignocellulosic biomass to VFAs and microbial protein symbiotically. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rDNA fragments revealed that the bacterial community was relatively stable after 1 week in cultivation, though it was different from the original community structure. Furthermore, sequence analysis of the DGGE bands indicates that the microbial community includes a cellulolytic bacterium, a bacterium acting synergistically with cellulolytic bacteria, and a propionate-producing bacterium, as well as other anaerobic bacteria. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Histamine Induces Bovine Rumen Epithelial Cell Inflammatory Response via NF-κB Pathway.

Science.gov (United States)

Sun, Xudong; Yuan, Xue; Chen, Liang; Wang, Tingting; Wang, Zhe; Sun, Guoquan; Li, Xiaobing; Li, Xinwei; Liu, Guowen

2017-01-01

Subacute ruminal acidosis (SARA) is a common disease in high-producing lactating cows. Rumenitis is the initial insult of SARA and is associated with the high concentrations of histamine produced in the rumen of dairy cows during SARA. However, the exact mechanism remains unclear. The objective of the current study is to investigate whether histamine induces inflammation of rumen epithelial cells and the underlying mechanism of this process. Bovine rumen epithelial cells were cultured and treated with different concentrations of histamine and pyrrolidine dithiocarbamate (PDTC, an NF-κB inhibitor) cultured in different pH medium (pH 7.2 or 5.5). qRT-PCR, Western-blotting, ELISA and immunocytofluorescence were used to evaluate whether histamine activated the NF-κB pathway and inflammatory cytokines. The results showed that histamine significantly increased the activity of IKK β and the phosphorylation levels of IκB α, as well as upregulated the mRNA and protein expression levels of NF-κB p65 in the rumen epithelial cells cultured in neutral (pH=7.2) and acidic (pH=5.5) medium. Furthermore, histamine treatment also significantly increased the transcriptional activity of NF-κB p65. High expression and transcriptional activity of NF-κB p65 significantly increased the mRNA expressions and concentrations of inflammatory cytokines, tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and interleukin 1 beta (IL-1β), thereby inducing the inflammatory response in bovine rumen epithelial cells. However, inhibition of NF-κB p65 by PDTC significantly decreased the expressions and concentrations of the inflammatory cytokines induced by histamine in the rumen epithelial cells cultured in the neutral and acidic medium. The present data indicate that histamine induces the inflammatory response of bovine rumen epithelial cells through the NF-κB pathway. © 2017 The Author(s). Published by S. Karger AG, Basel.

Mechanism of Excretion of a Bacterial Proteinase: Demonstration of Two Proteolytic Enzymes Produced by a Sarcina Strain (Coccus P)

Energy Technology Data Exchange (ETDEWEB)

SARNER, NITZA Z; BISSELL, MINA J; GIROLAMO, MARIO Di; GORINI, LUIGI

1970-06-29

A Sarcina strain (Coccus P) produces two proteolytic enzymes. One is found only extracellularly, is far more prevalent, and is actively excreted during exponential growth. It is the enzyme responsible for the known strong proteolytic activity of the cultures of this strain. A second protease is, however, produced which remains associated with the intact cells but is released by the protoplasts. The two enzymes appear unrelated in their derivation. Calcium ions play an essential role in preventing autodigestion of the excreted enzyme. Bacterial proteins are found outside the cell boundary as a consequence either of passive processes such as leakage or lysis or of active excretion. Under conditions in which leakage and lysis do not occur, as during exponential growth, the cell boundary is a barrier causing a complete separation of the bulk of the intracellular proteins from the one or very few extracellular proteins, with no trace of either type being detectable on the wrong side of the boundary. Since in bacteria there is no evidence of protein being produced other than internally, the separation into intraand extracellular proteins should occur after peptide chain formation. The question arises as to whether the structure of the cell boundary or that of the excreted proteins themselves determines this separation. Coccus P, a Sarcina closely related to Micrococcus lysodeikticus (3), produces an extracellular proteinase during the exponential phase of growth so that the process appears to be active excretion. The organism grows exponentially in a defined synthetic medium (12) to relatively high cell density (10{sup 9} cells/ml); therefore the mechanism of excretion can be studied over an extended period of time without the difficulties of changing growth rates. Coagulation of reconstituted skim milk provides a simple and sensitive assay for enzyme activity (I 1). The extracellular proteinase has also been purified and partially characterized (6-8). It has been shown

Alternatives for optimisation of rumen fermentation in ruminants

Directory of Open Access Journals (Sweden)

T. Slavov

2017-06-01

Full Text Available Abstract. The proper knowledge on the variety of events occurring in the rumen makes possible their optimisation with respect to the complete feed conversion and increasing the productive performance of ruminants. The inclusion of various dietary additives (supplements, biologically active substances, nutritional antibiotics, probiotics, enzymatic preparations, plant extracts etc. has an effect on the intensity and specific pathway of fermentation, and thus, on the general digestion and systemic metabolism. The optimisation of rumen digestion is a method with substantial potential for improving the efficiency of ruminant husbandry, increasing of quality of their produce and health maintenance.

Highly Promiscuous Oxidases Discovered in the Bovine Rumen Microbiome

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Lisa Ufarté

2018-05-01

Full Text Available The bovine rumen hosts a diverse microbiota, which is highly specialized in the degradation of lignocellulose. Ruminal bacteria, in particular, are well equipped to deconstruct plant cell wall polysaccharides. Nevertheless, their potential role in the breakdown of the lignin network has never been investigated. In this study, we used functional metagenomics to identify bacterial redox enzymes acting on polyaromatic compounds. A new methodology was developed to explore the potential of uncultured microbes to degrade lignin derivatives, namely kraft lignin and lignosulfonate. From a fosmid library covering 0.7 Gb of metagenomic DNA, three hit clones were identified, producing enzymes able to oxidize a wide variety of polyaromatic compounds without the need for the addition of copper, manganese, or mediators. These promiscuous redox enzymes could thus be of potential interest both in plant biomass refining and dye remediation. The enzymes were derived from uncultured Clostridia, and belong to complex gene clusters involving proteins of different functional types, including hemicellulases, which likely work in synergy to produce substrate degradation.

Shorten fungal treatment of lignocellulosic waste with additives to improve rumen degradability

NARCIS (Netherlands)

Kuijk, van S.J.A.; Sonnenberg, A.S.M.; Baars, J.J.P.; Hendriks, W.H.; Cone, J.W.

2014-01-01

Selective lignin degrading fungi can be used as pre-treatment to make cellulose in plant cell walls accessible for rumen microbes. According to previous studies, Ceriporiopsis subvermispora and Lentinula edodes can increase the in vitro rumen degradability of lignocellulosic biomass in 7 to 8 weeks.

Proteolytic activity of alkaliphilic, salt-tolerant actinomycetes from ...

African Journals Online (AJOL)

tolerant alkaliphilic. All the isolates need to be further studied for the ability of their potential protease enzyme production. Key words: Alkaliphilic actinomycetes, salt tolerant actinomycetes, desert soil, isolation, proteolytic activity.

[Proteolytic activity of IgG-antibodies of mice, immunized by calf thymus histones].

Science.gov (United States)

Kit, Iu Ia; Korniĭ, N; Kril', I Ĭ; Mahorivs'ka, I B; Tkachenko, V; Bilyĭ, R O; Stoĭka, R S

2014-01-01

The main goal of the study was to determine the ability of histones to induce production of the proteolytically active IgG-antibodies in BALB/c mice. In order to perform this study 8 mice were immunized with the fraction of total calf thymus histones. IgGs were isolated from the serum of the immunized and not immunized animals by means of precipitation with 33% ammonium sulfate, followed by affinity chromatography on protein G-Sepharose column. Histones, myelin basic protein (MBP), lysozyme, BSA, ovalbumin, macroglobulin, casein and cytochrome c served as substrates for determining the proteolytic activity. It was found that IgGs from the blood serum of immunized mice are capable of hydrolyzing histone H1, core histone and MBP. On the contrary, the proteolytic activity of IgGs from the blood serum of not immunized mice was not detected. The absence of proteolytical enzymes in the fraction of IgGs was proven by HPLC chromatography. High levels of proteolytic activity toward histones have been also detected in affinity purified IgGs from blood serum of patients with rheumatoid arthritis, but not in healthy donors. These data indicate that eukaryotic histones may induce production of protabzymes in mammals. The possible origin of these protabzymes and their potential biological role in mammalians is discussed.

Bicarbonate-dependent secretion and proteolytic processing of recombinant myocilin.

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José-Daniel Aroca-Aguilar

Full Text Available Myocilin is an extracellular glycoprotein of poorly understood function. Mutations of this protein are involved in glaucoma, an optic neuropathy characterized by a progressive and irreversible visual loss and frequently associated with elevated intraocular pressure. We previously showed that recombinant myocilin undergoes an intracellular proteolytic processing by calpain II which cleaves the central region of the protein, releasing one N- and one C-terminal fragment. Myocilin cleavage is reduced by glaucoma mutations and it has been proposed to participate in intraocular pressure modulation. To identify possible factors regulating the proteolytic processing of recombinant myocilin, we used a cellular model in which we analyzed how different culture medium parameters (i.e., culture time, cell density, pH, bicarbonate concentration, etc. affect the presence of the extracellular C-terminal fragment. Extracellular bicarbonate depletion associated with culture medium acidification produced a reversible intracellular accumulation of full-length recombinant myocilin and incremented its intracellular proteolytic processing, raising the extracellular C-terminal fragment percentage. It was also determined that myocilin intracellular accumulation depends on its N-terminal region. These data suggest that aqueous humor bicarbonate variations could also modulate the secretion and cleavage of myocilin present in ocular tissues.

Effect of rice straw silage treated with rumen microbes of buffalo on digestibility and ecosystem of cattle rumen

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Thalib A

2000-03-01

Full Text Available Treatment of rice straw silage with addition of buffalo rumen microbes was conducted to improve the ruminal digestion of rice straw in ongole cattle. Three fistulated cattles were each introduced to dietary treatment: I. Untreated rice straw (JPTP, II. Rice straw ensilaged with buffalo rumen microbes (SJPMR-Kr, and ID. Elephant grass (RG. All diets were formulated isonitrogeneous (14% crude protein and fed to animals over a period of 4 weeks. After 4 weeks of feeding trial, rwnen fluid of the animals were evaluated to digest its own basal diet (as substrate. The results show that cumulative gas production resulting from the substrate fermented (96 hours by rumen fluid from cattle fed diet II is 205% of the diet I and 151 % of the diet ID. Measurements of DMD of the substrates after the gas production procedure show the similar trend (ie. DM digestibilities for JPTP= 33%; SJPMR-Kr= 54% dan RG= 45%. Means of in sacco DMD (72 hours incubation confirm the results of gas production (ie. in sacco DM Digestibilities for JPTP= 35%; SJPMR-Kr= 44% and RG= 39%. All results described between treatments are highly significant different (P0.05, except for total VFA (ie. JPTP= 0.52 mg Inri; SJPMR-Kr= 3,37 mg Inri and RG= 3.15 mg Inri.

The effect of cation source and dietary cation-anion difference on rumen ion concentrations in lactating dairy cows.

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Catterton, T L; Erdman, R A

2016-08-01

Many studies have focused on the influence of dietary cation-anion difference (DCAD) on animal performance but few have examined the effect of DCAD on the rumen ionic environment. The objective of this study was to examine the effects of DCAD, cation source (Na vs. K), and anion source (Cl vs. bicarbonate or carbonate) on rumen environment and fermentation. The study used 5 rumen-fistulated dairy cows and 5 dietary treatments that were applied using a 5×5 Latin square design with 2-wk experimental periods. Treatments consisted of (1) the basal total mixed ration (TMR); (2) the basal TMR plus 340mEq/kg of Na (dry matter basis) using NaCl; (3) the basal TMR plus 340mEq/kg of K using KCl; (4) the basal TMR plus 340mEq/kg of Na using NaHCO3; and (5) the basal TMR plus 340mEq/kg of K using K2CO3. On the last day of each experimental period, rumen samples were collected and pooled from 5 different locations at 0, 1.5, 3, 4.5, 6, 9, and 12h postfeeding for measurement of rumen pH and concentrations of strong ions and volatile fatty acids (VFA). Dietary supplementation of individual strong ions increased the corresponding rumen ion concentration. Rumen Na was decreased by 24mEq/L when K was substituted for Na in the diet, but added dietary Na had no effect on rumen K. Rumen Cl was increased by 10mEq/L in diets supplemented with Cl. Cation source had no effect on rumen pH or total VFA concentration. Increased DCAD increased rumen pH by 0.10 pH units and increased rumen acetate by 4mEq/L but did not increase total VFA. This study demonstrated that rumen ion concentrations can be manipulated by dietary ion concentrations. If production and feed efficiency responses to DCAD and ionophores in the diet are affected by rumen Na and K concentrations, then manipulating dietary Na and K could be used either to enhance or diminish those responses. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Isolation of proteolytic bacteria from mealworm (Tenebrio molitor) exoskeletons to produce chitinous material.

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da Silva, Fernanda Kerche Paes; Brück, Dieter W; Brück, Wolfram M

2017-09-15

The use of insects as a source of protein is becoming an important factor for feeding an increasing population. After protein extraction for food use, the insect exoskeleton may offer the possibility for the production of added value products. Here, the aim was to isolate bacteria from the surface of farmed mealworms (Tenebrio molitor Linnaeus, 1758) for the production of chitinous material from insect exoskeletons using microbial fermentation. Isolates were screened for proteases and acid production that may aid deproteination and demineralisation of insects through fermentation to produce chitin. Selected isolates were used single-step (isolated bacteria only) or two-step fermentations with Lactobacillus plantarum (DSM 20174). Two-step fermentations with isolates from mealworm exoskeletons resulted in a demineralisation of 97.9 and 98.5% from deproteinated mealworm fractions. Attenuated total reflectance-Fourier- transform infrared spectroscopy analysis showed that crude chitin was produced. However, further optimisation is needed before the process can be upscaled. This is, to our knowledge, the first report using microbial fermentation for the extraction of chitin from insects. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

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

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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

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

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Henk J van Lingen

Full Text Available 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

New radionuclide method for determination of gastric content proteolytic activity. (Experiments)

International Nuclear Information System (INIS)

Sergienko, V.B.; Popova, L.V.; Khodarev, N.N.; Astashenkova, K.Yu.

1988-01-01

A possibility of the use of a radionuclide tubeless rapid method for measuring gastric content proteolytic activity (GCPA) with the help of a protein (gelatin) RP containing capsule was demonstrated in experiments in vitro. There was correlation between the time of dissolution of RP containing capsules and GCPA determined after Mett's method. Reference time intervals were established for normal, raised and lowered proteolytic activity. The method was shown to be physiological, simple and timesaving

Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. II. Protozoa population and diversity of bacterial communities.

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Martínez, M E; Ranilla, M J; Tejido, M L; Saro, C; Carro, M D

2010-08-01

Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of dietary characteristics on microbial populations and bacterial diversity. The purpose of the study was to assess how closely fermenters can mimic the differences between diets found in vivo. The 4 experimental diets contained forage to concentrate (F:C) ratios of 70:30 (high forage; HF) or 30:70 (high concentrate; HC) with either alfalfa hay (A) or grass hay (G) as the forage. Total bacterial numbers were greater in the rumen of sheep fed HF diets compared with those fed HC diets, whereas the opposite was found in fermenters. The numbers of cellulolytic bacteria were not affected by F:C ratio in any fermentation system, but cellulolytic numbers were 2.7 and 1.8 times greater in sheep than in fermenters for HF and HC diets, respectively. Neither total bacterial nor cellulolytic numbers were affected by the type of forage in sheep or fermenters. Decreasing F:C ratio increased total protozoa and Entodiniae numbers in sheep by about 29 and 25%, respectively, but it had no effect in fermenters. Isotrichidae and Ophryoscolecinae numbers in sheep were not affected by changing F:C ratio, but both disappeared completely from fermenters fed HC diets. Total protozoa and Entodiniae numbers were greater in sheep fed A diets than in those fed G diets, whereas the opposite was found in fermenters. Results indicate that under the conditions of the present study, protozoa population in Rusitec fermenters was not representative of that in the rumen of sheep fed the same diets. In addition, protozoa numbers in fermenters were 121 and 226 times lower than those in the sheep rumen for HF and HC diets, respectively. The automated ribosomal intergenic spacer analysis of the 16S ribosomal DNA was used to analyze the diversity of liquid- and solid-associated bacteria in both systems. A total of 170 peaks were detected in the automated ribosomal intergenic spacer analysis

Esubacute acidosis in rumen of high-yield dairy cows: Prevalence and prevention

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Petrujkić Branko T.

2008-01-01

Full Text Available The objective of the investigations presented in this paper was to establish the frequency of the incidence of subacute acidosis in the rumen of cows (SARA in the first three months of lactation and the possibilities for its prevention using a mineral mix based on bentonite, zeolite, magnesium oxide, and sodium bicarbonate (Mix plus. The values obtained for the rumen pH content show that subacute rumen acidosis occurs in in 20 percent of the examined cows in the early stage of lactation. For these investigations, cows in early stages of lactation were chosen and divided into 2 groups. Cows of the experimental group were administered a fodder mix which contained the mineral mix for a buffer effect (Mix plus. The average values of the rumen pH content in the control and the experimental group of cows at the beginning and on the 30th day of the experiment were approximately the same and did not differ significantly (p>0.05. On the 60th day of the experiment, the values for the electrochemical reaction of the rumen content for the control group amounted to an average of 6.219±0.18, and for the experimental group of cows it was 6.772±0.23. The obtained difference was statistically very significant (p<0.001. At the end of the experiment, on the 90th day, the average pH value of the rumen content of cows of the control group was 6.308±0.16, while this value in the experimental group of cows was significantly higher and amounted to 6.676±0.29 (p<0.01.

Quantitative estimation of net rates of production of bacterial and protozoal nitrogen and their interconversion in the rumen

International Nuclear Information System (INIS)

El-Fouly, H.A.

1983-01-01

A technique is described using 35 S-labelled bacteria or protozoa by which the rates of production of microbial and protozoal protein N may be calculated. The results indicate an average microbial protein yield of about 13.7gN.d -1 in sheep maintained on a diet consisting largely of cottonseed cake and wheat and rice bran. Evidence is presented that protozoa made little contribution to the microbial protein-N leaving the rumen. Also, the average rate of N flow from the protozoal to the bacterial pool was about 3.5g.d -1 , whereas about 1.4g.d -1 of bacterial N was consumed by protozoa. (author)

Histogenesis of rumen in one-humped camel (Camelus dromedarius)

OpenAIRE

E. Salimi Naghani* and L. Akradi1

2012-01-01

The aim of this study was to follow several sequence histological changes that occur during the histogenesis of the rumen in one-humped camel (Camelus dromedarius). Histogenesis study was carried out on 66 fetuses of camel from 50th day of gestation until birth (390 days), according to the most relevant histo-differentiation characteristics of the rumen in fetuses, these were divided into four groups: group I (5-24 cm crown-rump length (C-RL); 50-140 days); group II (24-30 cm C-RL; 140-160 d...

The Economic Effect of a Daily Supplementation of carob pods (Ceratonia siliqua L., on Rumen Fermentation and Lactating Goats Performance

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Ayman A. Hassan

2016-07-01

Full Text Available The present study was performed to investigate the effect of a daily supplementation of carob pods (Ceratonia siliqua L., on rumen fermentation and milk production of goats. Thirty two lactating does (weight ranged from 33�35 kg, aged 2-4 years old and from 2nd to 3th lactation season were randomly allocated into four similar groups (8 animals each. The animals were fed with isocaloric and isonitrogenous diets. Carob pods was daily supplemented at the rate of 0, 25, 50 or 100g /h/d. The lactating trial was extended for 75 days where goats were fed individually and fresh water was available at all time. Rumen fermentation parameters were monitored on three fistulated adult does. Results indicated that volatile fatty acids concentration, rumen volume, microbial protein synthesis and total bacteria counts were highest (P<0.05 with C50 group compared with other groups. While, ammonia-N concentration and protozoa count were lower (P<0.05 with C100 group compared with other groups. Milk production, protein and fat percentage were better (P<0.05 for C50 and C25 groups than those of C100 group. Supplementation of Carob pods at 50 g caused a marked (P<0.05 increase in the enzymatic antioxidant activity (SOD, CAT, GPx, and GSH but had a significant decrease (P<0.05 in TBARS compared to control group. Thus, it could be concluded that daily supplement of 50 g carob pods could be reasonable amount for goats performance without any adverse effect.

Effect of natural bioflavonoid on in vitro ruminal microbiota activity in sheep rumen liquor

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Moufida Aggoun

2017-05-01

Full Text Available A pure bioflavonoid (rutin was extracted from Eucalyptus globulus leaves and identified by Thin Layer Chromatography using purified flavonoids moieties as a control. The purified flavonoid was used in in vitro gas production test to evaluate its effect on rumen fermentation traits of three substrates: vetch-oat hay, alfalfa hay and wheat straw. The concentrations tested were at 0.5 and 1 mg/ml. Globally, the addition of rutin did not affect significantly gas production (P<0.001. Nevertheless, for both levels, rutin caused a slight decrease in methane production (P<0.05. The high reduction was observed for wheat straw (15.53%, 19.6% for 0.5mg/ml and 1mg/ml, respectively. However, in vitro degradability of the three substrates was increased but this increase was not statistically significant (P<0.001. There was not any significant change in PF and microbial biomass production due to the addition of rutin. At same, rutin inclusion did not affect ammonia production of alfalfa hay and vetch-oat hay, but that of wheat straw was significantly decreased (P<0.001. There was not any significant effect on the acetate : propionate ratio. This bioflavonoid has a potential to alter the rumen fermentation pattern, mainly, methane production. Thus, others studies will be conducted to evaluate the dose of administration which will have a maximum reduction in the methane emission and to establish its impact on ruminale microbiota composition especially protozoa and Archaea bacteria.

Failure of cellulolysis in the rumen of reindeer fed timothy silage

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Monica A. Olsen

1995-01-01

Full Text Available Three male reindeer (Rangifer tarandus tarandus calves were brought from mountain pastures in April and fed regrowth timothy (Phleum pratense silage with 76% leaves and 24.0% dry matter (DM ad libitum. The silage contained (on DM basis 25.4% cellulose, 12.0% crude protein and 19-6% water soluble carbohydrates. After an initial period of 11 days the daily silage intake rose to almost similar values for all animals, but independently of food intake, body mass (BM increased by as much as 13.3 kg for animal R3 during the first 21 days, compared to 4.4 kg and 2.8 kg for Rl and R2, respectively. At slaughter the wet weight of the rumen contents of animal R3 constituted 30.2% of the total BM, compared to 18.5% and 19.1% in animals Rl and R2, respectively. A reduced ability of the rumen micro-biota to ferment pure cellulose in vitro was observed in R3. The ruminal pH was 7.07 and the concentration of volatile fatty acids was only 50.0 mM in R3, indicating a low rate of fermentation. The initial rates of in vitro dry matter digestibility of timothy silage and standard hay were also affected by the rumen fermentation failure in animal R3. Depressed rumen cellulolysis, which may be related to natural periods of starvation prior to the feeding experiment, could have caused the low rate of fermentation and the large rumen size observed in this animal.

In vitro biodegradation of cyanotoxins in the rumen fluid of cattle.

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Manubolu, Manjunath; Madawala, Samanthi R P; Dutta, Paresh C; Malmlöf, Kjell

2014-05-08

In countries around the Baltic Sea grazing ruminants have access to and drink, surface water from lakes, rivers and in several coastal regions. The water quality of these naturally occurring reservoirs affects performance and health of livestock. In the Baltic Sea both microcystin (MC) and nodularin (NOD) occurs as cyclic peptides and have hepatotoxic effects. Although cattle obviously have died after consuming contaminated water very little information is available as to how susceptible ruminants are to the toxins produced by cyanobacteria. The critical question as to whether the rumen microflora might constitute a protective shield is unresolved. For this reason our aim is to investigate a possible degradation rate of these toxins in rumen. The ability of rumen microorganisms to degrade certain important cyanotoxins (MC-LR, YR, RR and NOD) was studied in vitro by incubating with rumen fluid at three different concentrations (0.05, 0.5 and 5 μg/mL) for 3 h. The degradation efficiencies were determined by LC-MS (ESI) positive mode. Degradation was observed in the following order MC-RR 36%, NOD 35%, MC-RR 25% and MC-LR 8.9% at lower concentrations within 3 h. However, average degradation was observed at concentration of 0.5 μg/mL. No degradation was observed in higher concentrations for entire 3 h. The present results reveal that the degradation was both dose and time dependent. In conclusion the present results suggest that the rumen microbial flora may protect ruminants from being intoxicated by Cyanotoxins.

EFFECT OF MARINATION WITH PROTEOLYTIC ENZYMES ON QUALITY OF BEEF MUSCLE

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Daniela Istrati

2012-03-01

Full Text Available During storage and thermal treatment meat suffers a number of biochemical and physical-chemical changes in the substrate protein, changes that take place with varying intensity depending on the method of preservation utilized and temperature of thermal treatment applied. Application of different treatments aimed to influence the proteolytic activity as is the case of enzymatic tenderization of beef.Improving the meat tenderness with proteolytic enzymes is promising, but current legislation restricting the use of proteolytic enzymes from bacterial origin and recommended tenderizers salts containing papain, ficin and bromelain. Recent research revealed that meat marinating before grilling results in a reduction of heterocyclic amine content after thermal treatment. Also, the addition of fruit pulp, garlic or other spices contributes to decreased production of heterocyclic amines because of their antioxidant activity. In the present study was aimed influence of exogenous proteolytic enzymes on adult beef tenderness. To increase the tenderness of adult beef were used exogenous enzymes preparations (papain and bromelain and natural sources of enzymes using pineapple and papaya fruit. It was intended to establish the correlation between enzymatic activity of enzymes used in the study, the processing technology and changes in the physical-chemical and biochemical characteristics that occur during storage in refrigerated conditions (evolution of the rigidity index and water holding capacity, cooking losses and cooking yield of the samples injected/marinated with enzymes.

A note on the flora and fauna in the rumen of steers fed a feedlot bloat-provoking ration and the effect of penicillin.

Science.gov (United States)

BRYANT, M P; ROBINSON, I M; LINDAHL, I L

1961-11-01

A study was made of the predominant culturable bacteria and ciliate protozoa present in the rumen of two steers that were regularly bloating on a pelleted ratio containing 22% alfalfa meal, 16% soybean oil meal, 61% barley, and 1% common salt. The ruminal microorganisms in the two animals differed as indicated by a high total culture count of bacteria, an almost complete absence of ciliate protozoa, a low pH, and a difference in the proportions of presumptively identified predominant bacterial groups in one animal (steer 26) as compared with the other (steer 32). The first exposure of the animals to procaine penicillin (75 or 150 mg per day on 2 successive days) resulted in an abnormal ruminal flora 31 hr after the first treatment as indicated by drastic drops in total and cellulolytic bacterial counts and a change in the proportions of predominant bacterial groups. The animals refused feed for 32 to 48 hr after the first treatment. After feed consumption resumed, further treatment with 75 mg penicillin on 4 successive days did not appear to greatly alter the flora and did not result in feed refusal in animal 32. In animal 26, amounts of penicillin progressing from 50 to 200 mg per day did not result in feed refusals and observations on rumen ingesta samples during this period indicated a decrease in total bacterial count, a great increase in numbers of ciliate protozoa, a higher pH, and a change in the proportions of predominant bacterial groups so that the ruminal picture was much more similar to that of animal 32 than to its own during the pre-penicillin period. Bloat was not relieved except during the period of feed refusal. The results indicate that the ruminal flora rapidly adapts to penicillin and that bloat of the feedlot type can occur in animals with widely differing numbers and kinds of bacteria and protozoa. Feedlot bloat does not appear to be correlated with the occurrence or numbers of any of the individual predominant groups of bacteria cultured.

Influence of Inoculum Content on Performance of Anaerobic Reactors for Treating Cattle Manure using Rumen Fluid Inoculum

OpenAIRE

Sunarso; S. Johari; I N. Widiasa; Budiyono

2009-01-01

Biogas productions of cattle manure using rumen fluid inoculums were determined using batch anaerobic digesters at mesophilic temperatures (room and 38.5 oC). The aim of this paper was to analyze the influence of rumen fluid contents on biogas yield from cattle manure using fluid rumen inoculums. A series of laboratory experiments using 400 ml biodigester were performed in batch operation mode. Given 100 grams of fresh cattle manure (M) was fed to each biodigester and mixed with rumen fluid (...

A metagenomics approach to evaluate the impact of dietary supplementation with Ascophyllum nodosum or Laminaria digitata on rumen function in rusitec fermenters

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Alejandro eBelanche

2016-03-01

Full Text Available 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

Effect of DNA extraction and sample preservation method on rumen bacterial population.

Science.gov (United States)

Fliegerova, Katerina; Tapio, Ilma; Bonin, Aurelie; Mrazek, Jakub; Callegari, Maria Luisa; Bani, Paolo; Bayat, Alireza; Vilkki, Johanna; Kopečný, Jan; Shingfield, Kevin J; Boyer, Frederic; Coissac, Eric; Taberlet, Pierre; Wallace, R John

2014-10-01

The comparison of the bacterial profile of intracellular (iDNA) and extracellular DNA (eDNA) isolated from cow rumen content stored under different conditions was conducted. The influence of rumen fluid treatment (cheesecloth squeezed, centrifuged, filtered), storage temperature (RT, -80 °C) and cryoprotectants (PBS-glycerol, ethanol) on quality and quantity parameters of extracted DNA was evaluated by bacterial DGGE analysis, real-time PCR quantification and metabarcoding approach using high-throughput sequencing. Samples clustered according to the type of extracted DNA due to considerable differences between iDNA and eDNA bacterial profiles, while storage temperature and cryoprotectants additives had little effect on sample clustering. The numbers of Firmicutes and Bacteroidetes were lower (P rumen fluid subjected to the eDNA isolation procedure considerably changed the ratio of molecular operational taxonomic units (MOTUs) of Bacteroidetes and Firmicutes. Intracellular DNA extraction using bead-beating method from cheesecloth sieved rumen content mixed with PBS-glycerol and stored at -80 °C was found as the optimal method to study ruminal bacterial profile. Copyright © 2013 Elsevier Ltd. All rights reserved.

Seasonal variation in the temperature sensitivity of proteolytic enzyme activity in temperate forest soils

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Brzostek, Edward R.; Finzi, Adrien C.

2012-03-01

Increasing soil temperature has the potential to alter the activity of the extracellular enzymes that mobilize nitrogen (N) from soil organic matter (SOM) and ultimately the availability of N for primary production. Proteolytic enzymes depolymerize N from proteinaceous components of SOM into amino acids, and their activity is a principal driver of the within-system cycle of soil N. The objectives of this study were to investigate whether the soils of temperate forest tree species differ in the temperature sensitivity of proteolytic enzyme activity over the growing season and the role of substrate limitation in regulating temperature sensitivity. Across species and sampling dates, proteolytic enzyme activity had relatively low sensitivity to temperature with a mean activation energy (Ea) of 33.5 kJ mol-1. Ea declined in white ash, American beech, and eastern hemlock soils across the growing season as soils warmed. By contrast, Eain sugar maple soil increased across the growing season. We used these data to develop a species-specific empirical model of proteolytic enzyme activity for the 2009 calendar year and studied the interactive effects of soil temperature (ambient or +5°C) and substrate limitation (ambient or elevated protein) on enzyme activity. Declines in substrate limitation had a larger single-factor effect on proteolytic enzyme activity than temperature, particularly in the spring. There was, however, a large synergistic effect of increasing temperature and substrate supply on proteolytic enzyme activity. Our results suggest limited increases in N availability with climate warming unless there is a parallel increase in the availability of protein substrates.

Effects of feed intake and dietary urea concentration on ruminal dilution rate and efficiency of bacteria growth in steers

International Nuclear Information System (INIS)

Firkins, J.L.; Lewis, S.M.; Montgomery, L.; Berger, L.L.; Merchen, N.R.; Fahey, G.C. Jr.

1987-01-01

Four multiple-fistulated steers (340 kg) were fed a diet containing 50% ground grass hay, 20% dry distillers grains, and 30% concentrate at two intakes (7.2 or 4.8 kg DM/d). Urea (.4 or 1.2% of the diet) was infused continuously into the steers' rumens. The experimental design was a 4 x 4 Latin square with a 2 x 2 factorial arrangement of treatments. Infusing urea at .4 or 1.2% of the diet resulted in ruminal NH 3 N concentration of 4.97 and 9.10 mg/dl, respectively. Feeding steers at high rather than low intake decreased ruminal and total tract digestibilities of organic matter, NDF, and ADF but did not increase ruminal escape of N. However, apparent N escape from the rumen calculated using purines, but not 15 N, as a bacterial marker was higher when 1.2 vs. .4% urea was infused. Feeding at high rather than at low intake increased the total pool of viable bacteria per gram organic matter fermented in the rumen. Although ruminal fluid outflows and particulate dilution rates were greater when steers were fed at high than low intakes, efficiencies of bacterial protein synthesis were unaffected by intake. The possibility of increased N recycling within the rumen with feeding at the higher intake is discussed

The rumen microbial metaproteome as revealed by SDS-PAGE.

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Snelling, Timothy J; Wallace, R John

2017-01-07

Ruminal digestion is carried out by large numbers of bacteria, archaea, protozoa and fungi. Understanding the microbiota is important because ruminal fermentation dictates the efficiency of feed utilisation by the animal and is also responsible for major emissions of the greenhouse gas, methane. Recent metagenomic and metatranscriptomic studies have helped to elucidate many features of the composition and activity of the microbiota. The metaproteome provides complementary information to these other -omics technologies. The aim of this study was to explore the metaproteome of bovine and ovine ruminal digesta using 2D SDS-PAGE. Digesta samples were taken via ruminal fistulae and by gastric intubation, or at slaughter, and stored in glycerol at -80 °C. A protein extraction protocol was developed to maximise yield and representativeness of the protein content. The proteome of ruminal digesta taken from dairy cows fed a high concentrate diet was dominated by a few very highly expressed proteins, which were identified by LC-MS/MS to be structural proteins, such as actin and α- and β-tubulins, derived from ciliate protozoa. Removal of protozoa from digesta before extraction of proteins revealed the prokaryotic metaproteome, which was dominated by enzymes involved in glycolysis, such as glyceraldehyde-3-phosphate dehydrogenase, phosphoenolpyruvate carboxykinase, phosphoglycerate kinase and triosephosphate isomerase. The enzymes were predominantly from the Firmicutes and Bacteroidetes phyla. Enzymes from methanogenic archaea were also abundant, consistent with the importance of methane formation in the rumen. Gels from samples from dairy cows fed a high proportion of grass silage were consistently obscured by co-staining of humic compounds. Samples from beef cattle and fattening lambs receiving a predominantly concentrate diet produced clearer gels, but the pattern of spots was inconsistent between samples, making comparisons difficult. This work demonstrated for the

Measurement of monoterpenes and sesquiterpenes in serum, plasma, and rumen fluid from sheep

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Studies involving the consumption, metabolism, and elimination of terpenes by small ruminants consuming terpene-laden shrubs as well as those exploring the potential for natural products as rumen modifiers could benefit from a procedure that measures terpenes in both blood and rumen fluid and that i...

Straw particle size in calf starters: Effects on digestive system development and rumen fermentation.

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Suarez-Mena, F X; Heinrichs, A J; Jones, C M; Hill, T M; Quigley, J D

2016-01-01

Two trials were conducted to determine effects of straw particle size in calf starter on rumen fermentation and development in calves. Holstein calves (n=17 in trial 1; n=25 in trial 2) were housed in individual pens; bedding (wood shavings) was covered with landscape fabric to completely avoid consumption of bedding. Milk replacer was fed at 12% of birth body weight per day and water offered free choice. Calves were randomly assigned to 4 treatments differing in geometric mean particle length (Xgm) of straw comprising 5% of starter dry matter. Straw was provided within the pellet at manufacture (PS; 0.82 mm Xgm) or mixed with the pellet at time of feeding at Xgm of 3.04 (SS), 7.10 (MS), or 12.7 (LS) mm. Calves (n=12; 3/treatment) in trial 1 were fitted with a rumen cannula by wk 2 of age. A fixed amount of starter that was adjusted with age and orts were fed through the cannula in cannulated calves. Calves were euthanized 6 wk after starter was offered (9 and 7 wk of age for trials 1 and 2, respectively). Rumen digesta pH linearly decreased with age, whereas volatile fatty acid concentration increased with age. Overall pH had a cubic trend with SS lower than that of PS and MS. Molar proportion of acetate decreased with age whereas propionate proportion increased. Overall molar proportions of volatile fatty acids were not affected by diet. Fecal Xgm was not different in spite of changes in diet particle size and rumen digesta of PS being greater than SS, MS, and LS at slaughter. Fecal pH and starch concentration were not affected by diet; however, pH decreased whereas starch content increased with age. Weight of stomach compartments, rumen papillae length and width, and rumen wall thickness did not differ between diets. Omasum weight as a percentage of body weight at harvest linearly decreased as straw particle size increased. Under the conditions of this study, modifying straw particle length in starter grain resulted in minimal rumen fermentation parameter

Evaluation of the contribution of recycled urea to the synthesis of the microbial of protein in the rumen using 15N labelled urea

International Nuclear Information System (INIS)

Al-Dehneh, A.M.

1990-01-01

Urea 15 N was continually infused into the juglar vein for 3 days in two duodenally cannulated cows fed diets of 1:2 and 2:1 forage: concentrate. Duodenal digesta samples were taken every 3 hours, and coccygeal blood and milk were sampled twice daily. Urine was collected for 5 days starting 1 day before infusion and total feces for 3 days during infusion. Fecal samples were also taken twice daily during the 5 days of collection. Urinary excretion of 15 N accounted for about 90% of that which existed from the body; whereas, feces and milk each accounted for about 5%. Recovery of 15 N during the infusion period ranged from 30 to 50% of that infused. Estimates using 15 N ratios, as percent of the total N passing into the duodenum, that was bacterial N, were 50% to 90% and appeared directly proportional to dry matter intake of cows. Recycled-N incorporated into rumen microbes was greater (24% verses 14% of N in bacteria passing into the duodenum) in cows fed the high concentrate than the high forage diet. Also, incorporation of recycled N into rumen microbes was higher in the lactating than the dry cow (24% verses 14%) and the flow of nitrogen from the rumen to the small intestine was greater for the concentrate than the forage diet (122.O% verses 101.0% of nitrogen intake). (author). 27 refs

SELEKSI DAN IDENTIFIKASI BAKTERI ENDOFIT POTENSIAL PENGHASIL ENZIM PROTEASE DARI TAMAN NASIONAL GUNUNG HALIMUN - (The Selection and Identification of Potential Endophyte Bacteria as Protease Enzyme Producer from Halimun Mount National Park

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Ruth Melliawati

2016-12-01

Full Text Available Endophytic bacteria have an equal chance to bacteria that live outside the plant tissue as potential bacteria. The selection has done towards 326 bacterial endophyte isolates. This research aimed to find and identify proteolytic potential isolates. The proteolytic selection of endophytic bacteria had done using solid skim milk. The capability of endophytic bacteria to agglomerate milk was tested using liquid skim milk which incubated for 7 days at room temperature. Enzyme production of four selected isolates was made through fermentation in GYS medium. The results showed that 86 isolates have proteolytic potential. Isolate HL.29B.63 had highest protease enzymes activity (65.918 U/mL. Medium optimization was able to increase the enzyme activity into 89.94% (125.04 U/mL. The analysis used 16s rDNA showed that isolate HL.29B.63 was Bacillus amyloliquefacient subs. plantarum strain FZB42.Keywords: endophytic bacteria, fermentation, identification, protease, selection ABSTRAKBakteri endofit mempunyai peluang yang sama dengan bakteri yang hidup diluar jaringan tanaman sebagai bakteri potensial. Seleksi dilakukan terhadap 326 isolat bakteri endofit. Tujuan penelitian ini adalah mencari isolat yang berpotensi proteolitik dan mengidentifikasinya. Seleksi proteolitik terhadap bakteri endofitik menggunakan skim milk padat. Uji kemampuan bakteri endofitik dalam menggumpalkan susu menggunakan medium skim milk cair yang diinkubasi selama 7 hari pada suhu ruang. Produksi enzim terhadap empat isolat terseleksi dilakukan melalui fermentasi dalam medium GYS. Hasilnya menunjukkan bahwa 86 isolat mempunyai potensi proteolitik. Isolat HL.29B.63 mempunyai aktif enzim protease tertinggi (65,918 U/mL. Optimasi medium dapat meningkatkan aktivitas enzim sebesar 89,94% (125,04 U/mL. Analisis menggunakan 16s rDNA menunjukkan bahwa isolat HL.29B.63 adalah Bacillus amyloliquefaciens subs. plantarum strain FZB42.Kata kunci: bakteri endofit, fermentasi, identifikasi, protease

Proteolytic Enzymes Clustered in Specialized Plasma-Membrane Domains Drive Endothelial Cells' Migration.

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Monica Salamone

Full Text Available In vitro cultured endothelial cells forming a continuous monolayer establish stable cell-cell contacts and acquire a "resting" phenotype; on the other hand, when growing in sparse conditions these cells acquire a migratory phenotype and invade the empty area of the culture. Culturing cells in different conditions, we compared expression and clustering of proteolytic enzymes in cells having migratory versus stationary behavior. In order to observe resting and migrating cells in the same microscopic field, a continuous cell monolayer was wounded. Increased expression of proteolytic enzymes was evident in cell membranes of migrating cells especially at sprouting sites and in shed membrane vesicles. Gelatin zymography and western blotting analyses confirmed that in migrating cells, expression of membrane-bound and of vesicle-associated proteolytic enzymes are increased. The enzymes concerned include MMP-2, MMP-9, MT1-MMP, seprase, DPP4 (DiPeptidyl Peptidase 4 and uPA. Shed membrane vesicles were shown to exert degradative activity on ECM components and produce substrates facilitating cell migration. Vesicles shed by migrating cells degraded ECM components at an increased rate; as a result their effect on cell migration was amplified. Inhibiting either Matrix Metallo Proteases (MMPs or Serine Integral Membrane Peptidases (SIMPs caused a decrease in the stimulatory effect of vesicles, inhibiting the spontaneous migratory activity of cells; a similar result was also obtained when a monoclonal antibody acting on DPP4 was tested. We conclude that proteolytic enzymes have a synergistic stimulatory effect on cell migration and that their clustering probably facilitates the proteolytic activation cascades needed to produce maximal degradative activity on cell substrates during the angiogenic process.

Proteolytic Enzymes Clustered in Specialized Plasma-Membrane Domains Drive Endothelial Cells' Migration.

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Salamone, Monica; Carfì Pavia, Francesco; Ghersi, Giulio

2016-01-01

In vitro cultured endothelial cells forming a continuous monolayer establish stable cell-cell contacts and acquire a "resting" phenotype; on the other hand, when growing in sparse conditions these cells acquire a migratory phenotype and invade the empty area of the culture. Culturing cells in different conditions, we compared expression and clustering of proteolytic enzymes in cells having migratory versus stationary behavior. In order to observe resting and migrating cells in the same microscopic field, a continuous cell monolayer was wounded. Increased expression of proteolytic enzymes was evident in cell membranes of migrating cells especially at sprouting sites and in shed membrane vesicles. Gelatin zymography and western blotting analyses confirmed that in migrating cells, expression of membrane-bound and of vesicle-associated proteolytic enzymes are increased. The enzymes concerned include MMP-2, MMP-9, MT1-MMP, seprase, DPP4 (DiPeptidyl Peptidase 4) and uPA. Shed membrane vesicles were shown to exert degradative activity on ECM components and produce substrates facilitating cell migration. Vesicles shed by migrating cells degraded ECM components at an increased rate; as a result their effect on cell migration was amplified. Inhibiting either Matrix Metallo Proteases (MMPs) or Serine Integral Membrane Peptidases (SIMPs) caused a decrease in the stimulatory effect of vesicles, inhibiting the spontaneous migratory activity of cells; a similar result was also obtained when a monoclonal antibody acting on DPP4 was tested. We conclude that proteolytic enzymes have a synergistic stimulatory effect on cell migration and that their clustering probably facilitates the proteolytic activation cascades needed to produce maximal degradative activity on cell substrates during the angiogenic process.

Proteolytic Enzymes Clustered in Specialized Plasma-Membrane Domains Drive Endothelial Cells’ Migration

Science.gov (United States)

Salamone, Monica; Carfì Pavia, Francesco

2016-01-01

In vitro cultured endothelial cells forming a continuous monolayer establish stable cell-cell contacts and acquire a “resting” phenotype; on the other hand, when growing in sparse conditions these cells acquire a migratory phenotype and invade the empty area of the culture. Culturing cells in different conditions, we compared expression and clustering of proteolytic enzymes in cells having migratory versus stationary behavior. In order to observe resting and migrating cells in the same microscopic field, a continuous cell monolayer was wounded. Increased expression of proteolytic enzymes was evident in cell membranes of migrating cells especially at sprouting sites and in shed membrane vesicles. Gelatin zymography and western blotting analyses confirmed that in migrating cells, expression of membrane-bound and of vesicle-associated proteolytic enzymes are increased. The enzymes concerned include MMP-2, MMP-9, MT1-MMP, seprase, DPP4 (DiPeptidyl Peptidase 4) and uPA. Shed membrane vesicles were shown to exert degradative activity on ECM components and produce substrates facilitating cell migration. Vesicles shed by migrating cells degraded ECM components at an increased rate; as a result their effect on cell migration was amplified. Inhibiting either Matrix Metallo Proteases (MMPs) or Serine Integral Membrane Peptidases (SIMPs) caused a decrease in the stimulatory effect of vesicles, inhibiting the spontaneous migratory activity of cells; a similar result was also obtained when a monoclonal antibody acting on DPP4 was tested. We conclude that proteolytic enzymes have a synergistic stimulatory effect on cell migration and that their clustering probably facilitates the proteolytic activation cascades needed to produce maximal degradative activity on cell substrates during the angiogenic process. PMID:27152413

Protein-energy supplementation for lambs: feed intake, ingestive behavior, rumen parameters and nutrient digestibility

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Pâmila Carolini Gonçalves da Silva

2017-08-01

Full Text Available The study evaluated the effects of dietary protein-energy supplementation on feed intake, ingestive behavior, rumen parameters and nutrient digestibility in lambs. Four castrated lambs with 31.9 kg mean body weight and fistulated rumen were tested. distributed into latin square design (4x4, four treatments were tested over four periods of time: no supplementation (control or with supplementation at 8, 16 and 24 g kg-1 body weight. The supplement (soybean meal, soybean hulls, ground corn and minerals was provided with roughage (Tifton Bermudagrass, Cynodon spp., hay, which was offered ad libitum once a day, at 8h00. In treatments receiving 0 (control, 8, 16 and 24 g kg-1 supplementation, dry matter intake was 685.26, 742.86, 842.51 and 1013.33 g day-1, crude protein intake was 80.18, 95.98, 118.64, 150.14 g day-1 and metabolizable energy intake 1.55, 1.91, 2.31 and 2.98 g day-1, respectively. Treatments receiving the highest supplementation levels spent less time with rumination and feeding and rested for longer (P < 0.05. Protein-energy supplementation level did not affect rumen parameters. Average rumen pH was 6.3 and rumen ammonia nitrogen 165 mg dL-1; both were affected by sampling time. Supplementation levels until 24 g kg-1 BW improves feed intake and nutrient digestibility linearly and changes ingestive behavior, lowering rumination time without affecting rumen parameters.

Cell wall content and rumen dry matter disappearance of γ-irradiated wood by-products

International Nuclear Information System (INIS)

Flachowsky, G.; Baer, M.; Zuber, S.; Tiroke, K.

1990-01-01

Spruce sawdust and barks of spruce, pine and larch were irradiated with various doses of γ-rays (0; 0.1; 0.25; 0.5; 1.0 and 2.0 MGy). Cell wall constituents and rumen dry-matter disappearance (incubation time: 48 h) were determined. γ-Irradiation significantly reduced neutral detergent fibre and acid detergent fibre content of all by-products. The crude lignin of the wood by-products was not significantly influenced by γ-irradiation. Rumen dry-matter loss of untreated sawdust was 5.6%, that of barks between 18.2 (pine) and 64.6% (spruce). γ-Irradiation significantly increased rumen dry-matter loss. Increased washout due to solubilization and particle breakdown was mainly responsible for the higher dry-matter losses in the rumen after irradiation. The results do not justify practical use because of the high dose of irradiation required. (author)

Rumen undegradable protein (RUP) and its intestinal digestibility after steam flaking of cereal grains

DEFF Research Database (Denmark)

Chrenková, M; Formelová, Z; Ceresnáková, Z

2018-01-01

While it is known that heat treatment of cereal grains generally improves the nutritional value for ruminants, simultaneous information on rumen degradability and intestinal digestibility of the rumen by-pass is scarce, especially for non-starch constituents. The effect of steam flaking at 90°C...... flaking on chemical composition of cereal grains (crude protein, acid detergent fibre, neutral detergent fibre, and starch) were observed. The protein fractions that are relevant to rumen degradability were significantly influenced by the steam flaking: the non-protein nitrogen fraction (A) was reduced (P...... hand, steam flaking markedly increased buffer insoluble but neutral detergent soluble protein fraction (B2) by 15–25% for all three cereal grains, whereas effects on B3 fraction were not significant. Steam flaking was also associated with an increase of the rumen undegradable protein fraction (C...

Detection of fiber-digesting bacteria in the forestomach contents of llamas (Lama glama by PCR

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María E Cerón Cucchi

Full Text Available The high fibrolytic activity and large biomass of strictly-anaerobic bacteria that inhabit the rumen makes them primarily responsible for the degradation of the forage consumed by ruminants. Llamas feed mainly on low quality fibrous roughages that are digested by an active and diverse microflora. The products of this fermentation are volatile fatty acids and microbial biomass, which will be used by the animals. The aim of this study was to detect the three major fiber-digesting anaerobic bacteria in the forestomach contents of llamas by PCR. In this study, we detected Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes in the forestomach contents of eight native llamas from Argentina.

Characterization of differentially expressed genes in calf rumen epithelium in response to weaning

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During weaning, rumen epithelial cell function must transition from a pre-ruminant to a true ruminant state for efficient nutrient absorption and metabolism. During this time, the rumen grows to represent from 30 to 70% of the capacity of the gut, directly impacting net efficiency of feed conversion...

The effect of starch, inulin, and degradable protein on ruminal fermentation and microbial growth in rumen simulation technique

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Xiang H. Zhao

2014-03-01

Full Text Available A rumen simulation technique apparatus with eight 800 mL fermentation vessels was used to investigate the effects of rumen degradable protein (RDP level and non-fibre carbohydrate (NFC type on ruminal fermentation, microbial growth, and populations of ruminal cellulolytic bacteria. Treatments consisted of two NFC types (starch and inulin supplemented with 0 g/d (low RDP or 1.56 g/d (high RDP sodium caseinate. No significant differences existed among dietary treatments in the apparent disappearance of dietary nutrients except for dietary N, which increased with increased dietary RDP (P<0.001. Compared with starch, inulin treatments reduced the molar proportion of acetate (P<0.001, the acetate:propionate ratio (P<0.001, and methane production (P=0.006, but increased the butyrate proportion (P<0.001. Increased dietary RDP led to increases in production of total volatile fatty acid (P=0.014 and methane (P=0.050, various measures of N (P≤0.046, and 16s rDNA copy numbers of Ruminococcus flavefaciens (P≤0.010. Non-fibre carbohydrate source did not affect daily microbial N flow regardless of dietary RDP, but ammonia N production was lower for inulin than for starch treatments under high RDP conditions (P<0.001. Compared with starch treatments, inulin depressed the copy numbers of Fibrobacter succinogenes in solid fraction (P=0.023 and R. flavefaciens in liquid (P=0.017 and solid fractions (P=0.007, but it increased the carboxymethylcellulase activity in solid fraction (P=0.045. Current results suggest that starch and inulin differ in ruminal volatile fatty acid fermentation but have similar effects on ruminal digestion and microbial synthesis in vitro, although inulin suppressed the growth of partial ruminal cellulolytic bacteria.

[Lactic acid bacteria proteinase and quality of fermented dairy products--A review].

Science.gov (United States)

Zhang, Shuang; Zhang, Lanwei; Han, Xue

2015-12-04

Lactic acid bacteria (LAB) could synthesize cell envelope proteinase with weak activity, which primarily degrades casein. In addition to its crucial role in the rapid growth of LAB in milk, LAB proteinases are also of industrial importance due to their contribution to the formation of texture and flavor of many fermented dairy products. The proteolytic system, properties of proteinase, the degradation product of casein and its effect on the quality of fermented dairy products were reviewed in this manuscript.

Increase of forage dryness induces differentiated anatomical response in the sheep rumen compartments.

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Scocco, Paola; Mercati, Francesca; Tardella, Federico Maria; Catorci, Andrea

2016-08-01

The aim of this study was to investigate how the Surface Enlargement Factor (SEF) and the epithelial keratinization degree of sheep rumen change in response to phytomass production, and to forage fiber and water content during the pasture vegetative cycle. The study used eighteen sheep nourished with dry hay and cereals during the winter season and with fresh hay during the pasture vegetative cycle. We collected samples from rumen indicative regions for two consecutive years characterized by different rainfall and pasture productivity values. We evaluated the densities (D) of rumen papillae to estimate the rumen SEF, and the keratinization percentage of the epithelial lining; these parameters showed differentiated modifications in the four ruminal analyzed compartments in response to pasture seasonal conditions. In addition, we performed Canonical Redundancy Analysis (RDA) on the "keratinization and SEF" matrix constrained by phytomass, water, and crude fiber contents of pasture at different time in the two considered years to highlight how rumen features answer to pasture conditions. Atrium (A) and ventral sac (VS) keratinization showed a strict positive correlation to crude fiber, while SEF of VS was positively related to phytomass and forage water content. The degree of keratinization of the rumen VS epithelium proved to be a useful parameter for evaluating anatomical variations in the short term period related to pasture features; in addition, its monitoring could be carried out through biopsy, thus avoiding the killing of animals. The study also leads to the application of the 3Rs (Replacement; Reduction; and Refinement). Microsc. Res. Tech. 79:738-743, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Role of rumen butyrate in regulation of nitrogen utilization and urea nitrogen kinetics in growing sheep.

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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 urea entry (synthesis) rate was reduced ( 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 rumen papillae protein turnover in growing lambs. Under the feeding conditions used and the rate of continuous butyrate infusion into the rumen in the present studies, butyrate does not affect overall N retention in growing sheep. However, butyrate may play a role in the redistribution of urea N fluxes in the overall scheme of N metabolism.

Natural and artificial feeding management before weaning promote different rumen microbial colonization but not differences in gene expression levels at the rumen epithelium of newborn goats.

Science.gov (United States)

Abecia, Leticia; Jiménez, Elisabeth; Martínez-Fernandez, Gonzalo; Martín-García, A Ignacio; Ramos-Morales, Eva; Pinloche, Eric; Denman, Stuart E; Newbold, C Jamie; Yáñez-Ruiz, David R

2017-01-01

The aim of this work was to evaluate the effect of feeding management during the first month of life (natural with the mother, NAT, or artificial with milk replacer, ART) on the rumen microbial colonization and the host innate immune response. Thirty pregnant goats carrying two fetuses were used. At birth one kid was taken immediately away from the doe and fed milk replacer (ART) while the other remained with the mother (NAT). Kids from groups received colostrum during first 2 days of life. Groups of four kids (from ART and NAT experimental groups) were slaughtered at 1, 3, 7, 14, 21 and 28 days of life. On the sampling day, after slaughtering, the rumen content was sampled and epithelial rumen tissue was collected. Pyrosequencing analyses of the bacterial community structure on samples collected at 3, 7, 14 and 28 days showed that both systems promoted significantly different colonization patterns (P = 0.001). Diversity indices increased with age and were higher in NAT feeding system. Lower mRNA abundance was detected in TLR2, TLR8 and TLR10 in days 3 and 5 compared to the other days (7, 14, 21 and 28). Only TLR5 showed a significantly different level of expression according to the feeding system, presenting higher mRNA abundances in ART kids. PGLYRP1 showed significantly higher abundance levels in days 3, 5 and 7, and then experienced a decline independently of the feeding system. These observations confirmed a highly diverse microbial colonisation from the first day of life in the undeveloped rumen, and show that the colonization pattern substantially differs between pre-ruminants reared under natural or artificial milk feeding systems. However, the rumen epithelial immune development does not differentially respond to distinct microbial colonization patterns.

Natural and artificial feeding management before weaning promote different rumen microbial colonization but not differences in gene expression levels at the rumen epithelium of newborn goats.

Directory of Open Access Journals (Sweden)

Leticia Abecia

Full Text Available The aim of this work was to evaluate the effect of feeding management during the first month of life (natural with the mother, NAT, or artificial with milk replacer, ART on the rumen microbial colonization and the host innate immune response. Thirty pregnant goats carrying two fetuses were used. At birth one kid was taken immediately away from the doe and fed milk replacer (ART while the other remained with the mother (NAT. Kids from groups received colostrum during first 2 days of life. Groups of four kids (from ART and NAT experimental groups were slaughtered at 1, 3, 7, 14, 21 and 28 days of life. On the sampling day, after slaughtering, the rumen content was sampled and epithelial rumen tissue was collected. Pyrosequencing analyses of the bacterial community structure on samples collected at 3, 7, 14 and 28 days showed that both systems promoted significantly different colonization patterns (P = 0.001. Diversity indices increased with age and were higher in NAT feeding system. Lower mRNA abundance was detected in TLR2, TLR8 and TLR10 in days 3 and 5 compared to the other days (7, 14, 21 and 28. Only TLR5 showed a significantly different level of expression according to the feeding system, presenting higher mRNA abundances in ART kids. PGLYRP1 showed significantly higher abundance levels in days 3, 5 and 7, and then experienced a decline independently of the feeding system. These observations confirmed a highly diverse microbial colonisation from the first day of life in the undeveloped rumen, and show that the colonization pattern substantially differs between pre-ruminants reared under natural or artificial milk feeding systems. However, the rumen epithelial immune development does not differentially respond to distinct microbial colonization patterns.

Determination of the retention time of feed in rumen using Sc-46 tracer

International Nuclear Information System (INIS)

Nadai, E.A. de; Castro, F.B. de; Barros Ferraz, E.S. de; Machado, P.F.; Nascimento Filho, V.F. do; Pessenda, L.C.R.; Sao Paulo Univ., Piracicaba

1988-01-01

The retention time of fibrous feed materials in rumen is of prime importance by providing parameters to evaluate its nutritional value. Some existing marker methods have been presenting serious problems such as physico-chemical alterations of feedstuff during the treatment and also marker migration in rumen conditions. In this study, the rare-earth element Scandium-46 has been applied to marking sugarcane bagasse physically processed. The data showed no significant migration of marker to liquid phase. The retention time in the rumen fistulated cow was 23.06 hours. Correlation coefficient of elapsed time after marking with logarithm of specific activity of ruminal content was 0.9986**. (author) [pt

Rumen passage kinetics of forage and concentrate derived fiber in dairy cows

DEFF Research Database (Denmark)

Krämer, Monika; Lund, Peter; Weisbjerg, Martin Riis

2013-01-01

, were used in a completely randomized block experiment. Treatments differed in forage type (corn silage versus grass silage) and forage:concentrate ratio (50:50 versus 75:25 on organic matter basis). Fiber passage kinetics were studied based on rumen evacuations and on marker excretion profiles in feces....... The forage type itself (corn silage and grass silage) rather than ration composition seemed to determine the total tract retention time of forage fiber......Rumen passage kinetics of forage and concentrate fiber were analyzed to determine intrinsic feed effects and extrinsic ration effects on the retention time of fiber in the rumen. Sixteen Danish Holstein cows (557 + 37 kg body weight, 120 + 21 days in milk, mean + SD), 8 fitted with ruminal cannulas...

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

Science.gov (United States)

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).

Evaluation of the contribution of recycled urea to the synthesis of the microbial of protein in the rumen using sup 15 N labelled urea

Energy Technology Data Exchange (ETDEWEB)

Al-Dehneh, A M [Michigan State Univ. (USA) Dept. of Animal Science

1990-01-01

Urea {sup 15}N was continually infused into the juglar vein for 3 days in two duodenally cannulated cows fed diets of 1:2 and 2:1 forage: concentrate. Duodenal digesta samples were taken every 3 hours, and coccygeal blood and milk were sampled twice daily. Urine was collected for 5 days starting 1 day before infusion and total feces for 3 days during infusion. Fecal samples were also taken twice daily during the 5 days of collection. Urinary excretion of {sup 15}N accounted for about 90% of that which existed from the body; whereas, feces and milk each accounted for about 5%. Recovery of {sup 15}N during the infusion period ranged from 30 to 50% of that infused. Estimates using {sup 15}N ratios, as percent of the total N passing into the duodenum, that was bacterial N, were 50% to 90% and appeared directly proportional to dry matter intake of cows. Recycled-N incorporated into rumen microbes was greater (24% verses 14% of N in bacteria passing into the duodenum) in cows fed the high concentrate than the high forage diet. Also, incorporation of recycled N into rumen microbes was higher in the lactating than the dry cow (24% verses 14%) and the flow of nitrogen from the rumen to the small intestine was greater for the concentrate than the forage diet (122.O% verses 101.0% of nitrogen intake). (author). 27 refs.

Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils.

Science.gov (United States)

Aanniz, Tarik; Ouadghiri, Mouna; Melloul, Marouane; Swings, Jean; Elfahime, Elmostafa; Ibijbijen, Jamal; Ismaili, Mohamed; Amar, Mohamed

2015-06-01

The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. subtilis (subsp. spizizenii (2) and subsp. inaquosurum (6)), B. amyloliquefaciens (subsp. amyloliquefaciens (4) and subsp. plantarum (4)), B. tequilensis (3), B. pumilus (3) and Bacillus sp. (19). Only six isolates (2.5%) belonged to the genus Aeribacillus represented by A. pallidus (4) and Aeribacillus sp. (2). In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively.

Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils

Directory of Open Access Journals (Sweden)

Tarik Aanniz

2015-06-01

Full Text Available The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240 thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5% represented by B. licheniformis (119, B. aerius (44, B. sonorensis (33, B. subtilis (subsp. spizizenii (2 and subsp. inaquosurum (6, B. amyloliquefaciens (subsp. amyloliquefaciens (4 and subsp. plantarum (4, B. tequilensis (3, B. pumilus (3 and Bacillus sp. (19. Only six isolates (2.5% belonged to the genus Aeribacillus represented by A. pallidus (4 and Aeribacillus sp. (2. In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively.

Activation of human monocytes by proteolytic fragments of gliadin

Czech Academy of Sciences Publication Activity Database

Jelínková, Lenka; Tučková, Ludmila; Cinová, Jana; Cimburek, Zdeněk; Tlaskalová, Helena

2003-01-01

Roč. 87, č. 1 (2003), s. 08 ISSN 0165-2478 Institutional research plan: CEZ:AV0Z5020903 Keywords : proteolytic * fragments * gliadin Subject RIV: EE - Microbiology, Virology Impact factor: 1.710, year: 2003

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

Directory of Open Access Journals (Sweden)

Xufeng Han

Full Text Available 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.

Extracellular proteolytic enzymes produced by human pathogenic Vibrio species

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Shin-Ichi eMiyoshi

2013-11-01

Full Text Available Bacteria in the genus Vibrio produce extracellular proteolytic enzymes to obtain nutrients via digestion of various protein substrates. However, the enzymes secreted by human pathogenic species have been documented to modulate the bacterial virulence. Several species including Vibrio cholerae and V. vulnificus are known to produce thermolysin-like metalloproteases termed vibriolysin. The vibriolysin from V. vulnificus, a causative agent of serious systemic infection, is a major toxic factor eliciting the secondary skin damage characterized by formation of the hemorrhagic brae. The vibriolysin from intestinal pathogens may play indirect roles in pathogenicity because it can activate protein toxins and hemagglutinin by the limited proteolysis and can affect the bacterial attachment to or detachment from the intestinal surface by degradation of the mucus layer. Two species causing wound infections, V. alginolyticus and V. parahaemolyticus, produce another metalloproteases so-called collagenases. Although the detailed pathological roles have not been studied, the collagenase is potent to accelerate the bacterial dissemination through digestion of the protein components of the extracellular matrix. Some species produce cymotrypsin-like serine proteases, which may also affect the bacterial virulence potential. The intestinal pathogens produce sufficient amounts of the metalloprotease at the small intestinal temperature; however, the metalloprotease production by extra-intestinal pathogens is much higher around the body surface temperature. On the other hand, the serine protease is expressed only in the absence of the metalloprotease.

Effects of sheep breed and soybean meal supplementation on rumen environment and degradation kinetics

NARCIS (Netherlands)

Lourenco, A.; Cone, J.W.; Fontes, P.; Dias-Da-Silva, A.

2013-01-01

The aim of this study was to evaluate if the in vivo digestibility and intake differences, observed in previous studies, between Ile-de-France (IF) and Churra-da-Terra-Quente (CTQ) sheep breeds, were due to rumen environment and degradability differences. The intake, digestibility, rumen environment

The effect of age on in sacco estimates of rumen dry matter and ...

African Journals Online (AJOL)

This study was conducted to determine whether rumen dry matter and crude protein degradability in calves aged 8-10 weeks differs from that in mature cows. Five Holstein bull calves were rumen-fistulated at six weeks of age and were used in consecutive weekly 24 h trials from 8-20 weeks of age. Dry matter and crude ...

A simple technique for measurement of pressure in the tympanitic rumen of cattle.

Science.gov (United States)

Turner, C B; Whyte, T D

1978-05-13

The construction and method of use of a simple device for the non-invasive measurement of intra-rumenal pressure is outlined. Results obtained from calves suffering from increased intra-rumenal pressure (bloat) are shown. The method is capable of quantifying pressures involved in bloat and could be used to augment the visual assessment of bloat scoring.

Detoksifikasi Mikotoksin Melalui Optimalisasi Fungsi Rumen dengan Pemberian Ragi (MYCOTOXIN DETOXIFICATION THROUGH OPTIMIZATION THE RUMEN FUNCTION BY YEAST

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Dadik Pantaya

2016-03-01

Full Text Available Mycotoxins are toxic metabolites produced by some fungal species commonly found in food and feed,particularly in cereals. In intensive production systems, dairy cattle are commonly fed with cereal-richdiets and, consequently, are more exposed to micotoxins. Besides, such diet is often associated with ahigher risk of rumen acidosis which can also affect the performance and the helath of animal. In addition,the efficacy of microbial detoxification can be reduced during acidosis. For instance, some authors observeda decrease in the number of protozoa that are responsible for the degradation of some mycotoxins. Anotherconsequence of acidosis is the potential modification of ruminal absorption of mycotoxins, which until nowhas received scarce attention. Yeast Saccharomyces cerevisiae, probiotic additives have been shown toreduce the post-feeding drop in rumen pH and to increase the number of ruminal protozoa. This effect canbe positive in reducing the absorption and toxicity of mycotoxins in ruminantia.

Health status of birds fed diets containing three differently processed discarded vegetable-bovine blood-rumen content mixtures.

Science.gov (United States)

Ekunseitan, D A; Balogun, O O; Sogunle, O M; Yusuf, A O; Ayoola, A A; Egbeyale, L T; Adeyemi, O A; Allison, I B; Iyanda, A I

2013-04-01

This study was conducted to determine the effect of feeding three differently processed mixtures on health status of broilers. A total of 1080 day-old Marshal broilers were fed; discarded vegetable-fresh bovine blood-fresh rumen digesta (P1), discarded vegetable-ensiled bovine blood-fresh rumen digesta (P2) and discarded vegetable-fresh bovine blood-ensiled rumen digesta (P3) at three levels of inclusion (0, 3 and 6%). Data on blood parameters was taken and were subjected to 3 x 3 factorial arrangements in a completely randomized design. Birds fed P1 had least values (p rumen digesta (P3) up to 6% level of inclusion.

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.

Science.gov (United States)

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

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

OpenAIRE

Kudva, Indira T; Stanton, Thaddeus B; Lippolis, John D

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; d...

Evaluation of incubated defatted rubber seed meal with sheep rumen liquor for Pangasius diet

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Muhammad Agus Suprayudi

2015-10-01

Full Text Available ABSTRACT The research evaluated the use of rubber seed meal (Hevea brasiliensis; RBS incubated with sheep rumen liquor as a subtitution of soybean meal in catfish Pangasionodon sp. diet. The fish was cultured for 40 days and fed with the experimental diet containing RBS at five different diet compositions regarding to soybean meal substitution level, i.e. 0% (control, 12%, 23%, 34%, and 44%. Feeding was done three times a day to satiation. No significant different was found on fish-protein retention and survival rate in all treatments. Based on the study result, the use of rubber-seed meal (Hevea brasiliensis; RBS incubated with sheep rumen liquor could substitute soybean meal in catfish Pangasionodon sp. diet. Keywords: Hevea brasiliensis, Pangasionodon sp., catfish, sheep rumen liquor, rubber seed meal  ABSTRAK Penelitian ini mengevaluasi penggunaan tepung bungkil biji karet (Hevea brasiliensis; TBBK yang diinkubasi dengan cairan rumen domba sebagai pengganti tepung bungkil kedelai pada pakan ikan patin Pangasionodon sp. Pemeliharaan ikan dilakukan selama 40 hari dengan pemberian lima komposisi pakan berbeda sesuai tingkat substitusi tepung bungkil kedelai oleh tepung bungkil karet. TBBK yang ditambahkan untuk mengganti bungkil kedelai adalah sebesar 0%, 12%, 23%, 34% dan 44%. Pemberian pakan dilakukan selama tiga kali sehari secara at satiation. Tidak ditemukan perbedaan signifikan (P>0,05 pada nilai retensi protein dalam tubuh dan kelangsungan hidup ikan uji pada semua perlakuan. Berdasarkan hasil penelitian, dapat disimpulkan bahwa tepung bungkil biji karet yang diinkubasi dengan cairan rumen domba dapat digunakan sebagai pengganti bungkil kedelai pada pakan ikan patin Pangasionodon sp. Kata kunci: Hevea brasiliensis, Pangasionodon sp., patin, rumen domba, tepung biji karet 

Optical Imaging of Matrix Metalloproteinase-7 Activity in Vivo Using a Proteolytic Nanobeacon

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Randy L. Scherer

2008-05-01

Full Text Available Matrix metalloproteinases (MMPs are extracellular proteolytic enzymes involved in tumor progression. We present the in vivo detection and quantitation of MMP7 activity using a specific near-infrared polymer-based proteolytic beacon, PB-M7NIR. PB-M7NIR is a pegylated polyamidoamine PAMAM-Generation 4 dendrimer core covalently coupled to a Cy5.5-labeled peptide representing a selective substrate that monitors MMP7 activity (sensor and AF750 as an internal reference to monitor relative substrate concentration (reference. In vivo imaging of tumors expressing MMP7 had a median sensor to reference ratio 2.2-fold higher than a that of a bilateral control tumor. Ex vivo imaging of intestines of multiple intestinal neoplasia (APCMin mice injected systemically with PB-M7NIR revealed a sixfold increase in the sensor to reference ratio in the adenomas of APCMin mice compared with control intestinal tissue or adenomas from MMP7-null Min mice. PB-M7NIR detected tumor sizes as small as 0.01 cm2, and the sensor to reference ratio was independent of tumor size. Histologic sectioning of xenograft tumors localized the proteolytic signal to the extracellular matrix; MMP7-overexpressing tumors displayed an approximately 300-fold enhancement in the sensor to reference ratio compared with nonexpressing tumor cells. In APCMin adenomas, the proteolytic signal colocalized with the endogenously expressed MMP7 protein, with sensor to reference ratios approximately sixfold greater than that of normal intestinal epithelium. PB-M7NIR provides a useful reagent for the in vivo and ex vivo quantitation and localization of MMP-selective proteolytic activity.

Macrophage-mediated proteolytic remodeling of the extracellular matrix in atherosclerosis results in neoepitopes

DEFF Research Database (Denmark)

Skjøt-Arkil, Helene; Barascuk, Natasha; Register, Thomas

2010-01-01

in almost all stages of atherosclerosis, by both initiating atherosclerotic plaques and degrading them through the secretion of proteolytic enzymes leading to rupture. This review summarizes the literature on the role of macrophages and their proteolytic activity on proteins in the extracellular matrix (ECM......) of the atherosclerotic plaque with a view to suggest a novel approach for identification of vulnerable plaques and turnover by the use of a new type of biomarker. The PubMed database was searched using the terms macrophages, foam cells, atherosclerosis, CVD, ECM remodeling, biomarker, neoepitope, matrix...... of the constituents of the ECM of the atherosclerotic plaque. At present it is not clear which proteases play pivotal roles at distinct stages of pathogenesis, rather that the combined proteolytic potential with some proteases at early stages and other at later stages may result in plaque rupture. This macrophage...

Conjugated fatty acids and methane production by rumen microbes when incubated with linseed oil alone or mixed with fish oil and/or malate.

Science.gov (United States)

Li, Xiang Z; Gao, Qing S; Yan, Chang G; Choi, Seong H; Shin, Jong S; Song, Man K

2015-08-01

We hypothesized that manipulating metabolism with fish oil and malate as a hydrogen acceptor would affect the biohydrogenation process of α-linolenic acid by rumen microbes. This study was to examine the effect of fish oil and/or malate on the production of conjugated fatty acids and methane (CH4 ) by rumen microbes when incubated with linseed oil. Linseed oil (LO), LO with fish oil (LO-FO), LO with malate (LO-MA), or LO with fish oil and malate (LO-FO-MA) was added to diluted rumen fluid, respectively. The LO-MA and LO-FO-MA increased pH and propionate concentration compared to the other treatments. LO-MA and LO-FO-MA reduced CH4 production compared to LO. LO-MA and LO-FO-MA increased the contents of c9,t11-conjugated linoleic acid (CLA) and c9,t11,c15-conjugated linolenic acid (CLnA) compared to LO. The content of malate was rapidly reduced while that of lactate was reduced in LO-MA and LO-FO-MA from 3 h incubation time. The fold change of the quantity of methanogen related to total bacteria was decreased at both 3 h and 6 h incubation times in all treatments compared to the control. Overall data indicate that supplementation of combined malate and/or fish oil when incubated with linseed oil, could depress methane generation and increase production of propionate, CLA and CLnA under the conditions of the current in vitro study. © 2015 Japanese Society of Animal Science.

Voluntary intake, nitrogen metabolism and rumen fermentation ...

African Journals Online (AJOL)

Voluntary intake, nitrogen metabolism and rumen fermentation patterns in sheep given cowpea, silverleaf desmodium and fine-stem stylo legume hays as ... utilisation, the negative nitrogen retentions might indicate the inadequacy of the specific legume hays used as nitrogen supplementary feeds to sheep fed a basal diet

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

Science.gov (United States)

Kudva, Indira T; Stanton, Thaddeus B; Lippolis, John D

2014-02-21

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. 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. 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.

The epithelial sodium channel γ-subunit is processed proteolytically in human kidney

DEFF Research Database (Denmark)

Langkilde, Rikke Zachar; Skjødt, Karsten; Marcussen, Niels

2015-01-01

The epithelial sodium channel (ENaC) of the kidney is necessary for extracellular volume homeostasis and normal arterial BP. Activity of ENaC is enhanced by proteolytic cleavage of the gamma-subunit and putative release of a 43-amino acid inhibitory tract from the gamma-subunit ectodomain. We......ENaC was detected consistently only in tissue from patients with proteinuria and observed in collecting ducts. In conclusion, human kidney gammaENaC is subject to proteolytic cleavage, yielding fragments compatible with furin cleavage, and proteinuria is associated with cleavage at the putative prostasin...

Changes in Metabolically Active Bacterial Community during Rumen Development, and Their Alteration by Rhubarb Root Powder Revealed by 16S rRNA Amplicon Sequencing.

Science.gov (United States)

Wang, Zuo; Elekwachi, Chijioke; Jiao, Jinzhen; Wang, Min; Tang, Shaoxun; Zhou, Chuanshe; Tan, Zhiliang; Forster, Robert J

2017-01-01

The objective of this present study was to explore the initial establishment of metabolically active bacteria and subsequent evolution in four fractions: rumen solid-phase (RS), liquid-phase (RL), protozoa-associated (RP), and epithelium-associated (RE) through early weaning and supplementing rhubarb root powder in 7 different age groups (1, 10, 20, 38, 41, 50, and 60 d) during rumen development. Results of the 16S rRNA sequencing based on RNA isolated from the four fractions revealed that the potentially active bacterial microbiota in four fractions were dominated by the phyla Proteobacteria, Firmicutes , and Bacteroidetes regardless of different ages. An age-dependent increment of Chao 1 richness was observed in the fractions of RL and RE. The principal coordinate analysis (PCoA) indicated that samples in four fractions all clustered based on different age groups, and the structure of the bacterial community in RE was distinct from those in other three fractions. The abundances of Proteobacteria decreased significantly ( P < 0.05) with age, while increases in the abundances of Firmicutes and Bacteroidetes were noted. At the genus level, the abundance of the predominant genus Mannheimia in the Proteobacteria phylum decreased significantly ( P < 0.05) after 1 d, while the genera Quinella, Prevotella, Fretibacterium, Ruminococcus, Lachnospiraceae NK3A20 group , and Atopobium underwent different manners of increases and dominated the bacterial microbiota across four fractions. Variations of the distributions of some specific bacterial genera across fractions were observed, and supplementation of rhubarb affected the relative abundance of various genera of bacteria.

Effects of including NaOH-treated corn straw as a substitute for wheat hay in the ration of lactating cows on performance, digestibility, and rumen microbial profile.

Science.gov (United States)

Jami, E; Shterzer, N; Yosef, E; Nikbachat, M; Miron, J; Mizrahi, I

2014-03-01

This study measured the effects of including 5% NaOH-treated corn straw (T-CS) as a substitute for 15% wheat hay in the control total mixed ration (TMR) of lactating cows on performance, digestibility, and rumen microbial profile. Two groups of 21 cows each, similar in initial performance, were fed individually 1 of the 2 TMR examined. Voluntary dry matter intake of cows fed the control TMR was 4.3% higher than that of the T-CS cows, but in vivo dry matter and organic matter digestibilities of both groups were similar. Crude protein digestibility was higher in the control cows but digestibility of neutral detergent fiber polysaccharides (cellulose and hemicelluloses) was higher in the T-CS TMR. This was followed by 4.6% reduction in rumination time of the T-CS group. A slightly higher milk yield was observed in the control cows compared with the T-CS group; however, milk fat and milk protein content were higher in cows fed the T-CS TMR. This was reflected in 1.3% increase in energy-corrected milk yield and 5.34% increase in production efficiency (energy-corrected milk yield/intake) of the T-CS cows compared with the control. Welfare of the cows, as assessed by length of daily recumbence time, was improved by feeding the T-CS TMR relative to the control group. As a whole, the rumen bacterial community was significantly modulated in the T-CS group in the experimental period compared with the preexperimental period, whereas the bacterial community of the control group remained unchanged during this period. Out of the 8 bacterial species that were quantified using real-time PCR, a notable decrease in cellulolytic bacteria was observed in the T-CS group, as well as an increase in lactic acid-utilizing bacteria. These results illustrate the effect of T-CS on the composition of rumen microbiota, which may play a role in improving the performance of the lactating cow. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The effects of high levels of rumen degradable protein on rumen pH and histamine concentrations in dairy cows

NARCIS (Netherlands)

Pilachai, R.; Schonewille, J.T.; Thamrongyoswittayakul, C.; Aiumlamai, S.; Wachirapakom, C.; Everts, H.; Hendriks, W.H.

2012-01-01

An experiment was conducted to test the hypothesis that the supplementation of crude protein (CP) results in rumen acidosis and increased histamine concentrations in dairy cows. Six ruminally fistulated, non-pregnant dry cows were fed three experimental rations in a replicated 3 × 3 Latin square

Effects of yeast culture supplement on digestion of nutrients and rumen fermentation in cattle fed on grass silage barley diet

Directory of Open Access Journals (Sweden)

Pekka Huhtanen

1991-12-01

Full Text Available The effects of including yeast culture (Saccharomyces cerevisiae plus growth medium; 5 x 106 organisms/g on the digestion of dietary constituents in the rumen and total digestive tract were studied in a pair of monozygote twin bulls. The animals were fitted with cannulae in the rumen and in the proximal duodenum. A diet of grass silage, barley and rapeseed meal (445, 445 and 90 g/kg total dry matter (DM was fed, with and without addition of 10 g per day of yeast culture (YC, in two treatment sequences. The addition of YC had no effect on the mean values of rumen pH, ammonia N concentration or molar proportions of volatile fatty acids. Also, the postprandial changes in rumen fermentation pattern were similar when the diet did and did not contain the YC supplement. The peak concentration of lactic acid 1 h after feeding tended to be higher in cattle receiving the YC diet (13.9 v 6.0 mmol/l. Apparent digestibility of organic matter (OM (mean 0.780 and the proportion of OM digestion occurring in the rumen (mean 0.603 were not affected by YC. Likewise, there was no effect on rumen or total digestion of cell wall carbohydrates, and the results for the degradation of hay DM in the rumen and for particle-associated carboxymethylcellulase and xylanase activities indicated that YC had no effect on the rumen environment that could affect fibre digestion. Supplemental yeast did not affect the rate of microbial N synthesis (28.0 and 28.6 g/kg OM apparently digested in the rumen. The results indicate that the addition of YC to the diet is not likely to improve the efficiency of digestion and fermentation in the rumen of cattle given a diet based on grass silage and barley.

Effect of technical cashew nut shell liquid on rumen methane emission and lactation performance of dairy cows.

Science.gov (United States)

Branco, A F; Giallongo, F; Frederick, T; Weeks, H; Oh, J; Hristov, A N

2015-06-01

Technical-grade cashew nut shell liquid (TCNSL) is a by-product of the cashew nut industry in tropical countries, and is known to exhibit a wide range of biological activities, including inhibitory effect against gram-positive bacteria. This study was conducted to investigate the effects of TCNSL (73.3% cardanol, 16.4% cardol, and 3.0% methylcardol) on rumen methane emission, nutrient digestibility, dry matter intake, and milk yield and composition in dairy cows. Eight multiparous Holstein cows were used in a crossover design trial with two 21-d experimental periods. The diet was based on corn silage and alfalfa haylage and was formulated to meet or exceed the energy and metabolizable protein requirements of the cows. Treatments were control (no TCNSL supplementation) or 30 g/cow per day of TCNSL. Rumen carbon dioxide emission was not affected by TCNSL. Treatment had no effect on methane emission (542 vs. 511±35.3 g/cow per day, respectively) and methane emission intensity (15.0 vs. 13.9±0.58 g/kg of energy-corrected milk, respectively) and tended to decrease methane emission per kilogram of dry matter intake (20.2 vs. 18.6±1.04 g/kg, respectively). Dry matter intake (average 26.9±1.00 kg/d), milk yield (40.0±1.73 kg/d), and milk composition were not different between treatments. The TCNSL had no effect on N losses in urine and feces and total-tract apparent digestibility of nutrients, except digestibility of neutral detergent fiber tended to be increased compared with the control. Plasma urea and glucose concentrations were not affected by TCNSL. Concentration of milk C18:0 tended to be decreased (17%) by TCNSL compared with the control. In this study, TCNSL did not alter absolute methane emission in the rumen, but tended to decrease it by 8% per kilogram of dry matter intake. The TCNSL had no effect on milk yield and composition in dairy cows. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Determination of proteolytic activity using L-[4,5-3H]leucine-labelled globin as a substrate

International Nuclear Information System (INIS)

Maliopoulou, T.B.; Dionyssiou-Asteriou, A.; Loucopoulos, D.

1980-01-01

A simple and sensitive method for the assay of proteolytic enzyme activity is described. This is based on the digestion of L-[4,5- 3 H]leucine globin by proteolytic enzymes and radioactivity measurement of the trichloroacetic acid soluble cleavage products. (Auth.)

Experimental acute rumen acidosis in sheep: consequences on clinical, rumen, and gastrointestinal permeability conditions and blood chemistry.

Science.gov (United States)

Minuti, A; Ahmed, S; Trevisi, E; Piccioli-Cappelli, F; Bertoni, G; Jahan, N; Bani, P

2014-09-01

Acute acidosis was induced in sheep, and gastrointestinal permeability was assessed by using lactulose as a permeability marker. Metabolism was evaluated by monitoring blood metabolites. Four rams (72.5 ± 4.6 kg BW) were used in a 2 × 2 changeover design experiment. The experimental period lasted 96 h from -24 to 72 h. After 24 h of fasting (from -24 to 0 h) for both controls and acidosis-induced rams (ACID), 0.5 kg of wheat flour was orally dosed at 0 and 12 h of the experimental period to ACID, while the basal diet (grass hay, ad libitum) was restored to control. At 24 h, a lactulose solution (30 g of lactulose in 200 mL of water) was orally administered. Blood samples were collected at -24, 0, 24, 48, and 72 h of the experimental periods for the analysis of metabolic profiles and during the 10 h after lactulose dosage to monitor lactulose changes in blood. In addition, rumen and fecal samples were collected at 24 h of the experimental period. The acidotic challenge markedly reduced (P < 0.01) rumen pH and VFA but increased rumen d- and l-lactic acid (P < 0.01). Concurrently, a decrease of fecal pH and VFA occurred in ACID (P < 0.01), together with an abrupt increase (P < 0.01) of lactate and fecal alkaline phosphatase. Blood lactulose was significantly increased in ACID peaking 2 h after lactulose dosage. Blood glucose, β-hydroxybutyrate, Ca, K, Mg, and alkaline phosphatase showed a significant reduction (P < 0.05) at 24 h, whereas urea and NEFA declined (P < 0.05) from 48 to 72 h. A strong inflammatory acute phase response with oxidative stress in ACID group was observed from 24 to 72 h; higher values of haptoglobin (P < 0.01) were measured from 24 to 72 h and of ceruloplasmin from 48 (P < 0.05) to 72 h (P < 0.01). Among the negative acute phase reactants, plasma albumin, cholesterol, paraoxonase, and Zn concentration also decreased (P < 0.05) in ACID at different time points between 24 and 72 h after acidotic challenge start. A rise (P < 0.05) of reactive

Comparison of the production rates of protozoa in the rumen estimated by using labelled live and formaldehyde treated mixed protozoal cells as marker

International Nuclear Information System (INIS)

Verma, D.N.; Singh, U.B.

1979-01-01

14 C-labelled mixed protozoal cells of rumen origin, treated with formaldehyde to protect their metabolism in the rumen, were used to estimate protozoa production in the rumen, and comparison was made of the growth obtained by injecting live labelled mixed rumen protozoal cells used earlier. (auth.)

Presence and species identity of rumen flukes in cattle and sheep in the Netherlands.

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Ploeger, H W; Ankum, L; Moll, L; van Doorn, D C K; Mitchell, G; Skuce, P J; Zadoks, R N; Holzhauer, M

2017-08-30

The purpose of the study was to gain knowledge about the prevalence and identity of rumen flukes (RF) in cattle and sheep in the Netherlands. Routine faecal examinations of diagnostic submissions between May 2009 and September 2014 showed a mean annual herd or flock RF prevalence of 15.8% for cattle and 8.0% for sheep. Prevalence in cattle was higher after 2012 than before, which may reflect a change in detection method as well as an increase in true prevalence. During November and December 2014, an abattoir survey was conducted to allow for scoring of rumen fluke burden and to obtain specimens for molecular species characterization. Over 8 visits to 5 abattoirs in areas deemed to pose a high risk for trematode infection, 116 cows and 41 sheep from 27 herds and 10 flocks were examined. Prevalence of RF was higher in beef cattle than in dairy cattle and higher in cattle than in sheep. Median fluke burden was >100 specimens per animal for most positive animals. Using a semi-quantitative RF density score as a gold standard, sensitivity and specificity of a modified quantitative Dorsman egg counting method were estimated at 82.6% and 83.3%, respectively. Of 14 collected adult rumen flukes, twelve (8 bovine and 4 ovine specimens) were identified as Calicophoron daubneyi. The other two, of bovine origin, were identified as Paramphistomum leydeni, which was unexpected as in other European countries all recently collected rumen flukes in both cattle and sheep were identified as C. daubneyi. The findings implicate that multiple rumen fluke species, intermediate host species and transmission cycles may play a role in rumen fluke infections in the Netherlands. Copyright © 2017. Published by Elsevier B.V.

Factors affecting proteolytic action of Lactococcus lactis in cheese

NARCIS (Netherlands)

Youssef, Y.B.

1992-01-01

Model cheeses were developed to study the behaviour of proteolytic agents involved in cheese maturation under conditions that closely resemble those in normal cheese. The models were applied to study protein breakdown by Lactococcus lactis ssp. cremoris HP , as a

In vitro studies on magnesium uptake by rumen epithelium using magnesium-28

International Nuclear Information System (INIS)

Martens, H.; Harmeyer, J.; Breves, G.

1976-01-01

Magnesium-28 transfer across the rumen epithelium has been studied using surviving epithelia in an in vitro system. Net absorption of magnesium in the direction from lumen to blood could be observed as the result of two opposite unidirectional fluxes of different magnitude. Net uptake of magnesium occurred against an electrical potential difference, and was associated with the presence of an unaltered transmural potential difference in the mucosal tissue. Both the net transfer of magnesium and the transmural potential difference decreased during two hours of incubation. Unidirectional fluxes of magnesium and net efflux from the lumen were markedly reduced although not completely inhibited by the addition of ouabain (10 -4 mol/l). The findings suggest that the mechanism of magnesium absorption by the rumen epithelium can be considered as an active transport process, and that the rumen is the main area of magnesium absorption in the living animal. (author)

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

Directory of Open Access Journals (Sweden)

Gemma Henderson

Full Text Available 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

Effects of forage provision to young calves on rumen fermentation and development of the gastrointestinal tract.

Science.gov (United States)

Castells, L; Bach, A; Aris, A; Terré, M

2013-08-01

Fifteen Holstein male calves were randomly assigned to 1 of 3 dietary treatments according to age and body weight (BW) to determine the effects of feeding different forages sources on rumen fermentation and gastrointestinal tract (GIT) development. Treatments consisted of a starter (20% crude protein, 21% neutral detergent fiber) fed alone (CON) or supplemented with alfalfa (AH) or with oat hay (OH). All calves received 2L of milk replacer (MR) at 12.5% dry matter twice daily until 49 d of age. Calves received 2L of the same MR from 50 to 56 d of age and were weaned at 57 d of age. Individual starter, forage, and MR intakes were recorded daily and BW was recorded weekly. A rumen sample was taken weekly to determine rumen pH and volatile fatty acid concentrations. Three weeks after weaning, animals were harvested and each anatomical part of the GIT was separated and weighed with and without contents. Rumen pH was lower in CON than in OH and AH calves. Furthermore, acetate proportion in the rumen liquid tended to be greater in AH than in CON and OH treatments. Total GIT weight, expressed as a percentage of BW, tended to be greater in AH compared with the other 2 treatments. Rumen tissue tended to weigh more in CON than in OH animals. Animals with access to forage tended to have a greater expression of monocarboxylate transporter 1 than CON calves. In conclusion, calves supplemented with oat hay have a better rumen environment than calves offered no forage and do not have an increased gut fill. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Linkage of microbial ecology to phenotype: correlation of rumen microbial ecology to cattle's feed efficiency.