Fungi can improve stover digestibility due to their ability to secrete oxidative enzymes that depolymerize lignin, allowing the rumen microorganisms to access the polysaccharides of the plant cell wall. Some ascomycetes have shown good delignification capability; however, they have been scarcely evaluated for their ability to improve corn stover (CS) ruminal digestibility. We evaluated the laccase induction by CS of the CMU-196 strain of the ascomycete fungus Didymosphaeria sp. (syn. = Paraconiothyrium sp.). Also, we analyzed the capacity of such strain to modify the cell wall of CS and to improve its digestion by the ruminal microbiota. The CMU-196 strain showed a maximum extracellular laccase activity of 39.74 ± 0.24 U/L when an aqueous stover extract (SE, 10% v/v) was added to the growth medium. The addition of ground stover (GS, 2% w/v) increased the activity to a maximum of 262.27 ± 0.58 U/L. In solid-state fermentation (SSF) assays of GS, the strain degrades cell walls, destabilizing the vessels and tracheids of plant biomass; the protein content reaches a maximum of 33.2 g/kg dry matter (DM) at 70 days, while the crude fiber content shows the highest level of 314 g/kg DM at 14 days. SSF treatment of the CS increased the in vitro ruminal production of gas in a fraction that was considered nondigestible at 18 h, and gas production increased by 14% with respect to the untreated GS at 14 days. The CMU-196 strain can digest the plant cell wall and improve ruminal CS digestibility at a level equivalent to several basidiomycete species.
- MeSH
- Ascomycota enzymologie růst a vývoj metabolismus MeSH
- bachor mikrobiologie MeSH
- biomasa MeSH
- buněčná stěna metabolismus ultrastruktura MeSH
- fermentace MeSH
- krmivo pro zvířata analýza mikrobiologie MeSH
- kukuřice setá metabolismus ultrastruktura MeSH
- lakasa metabolismus MeSH
- lignin metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The way that fluids and particles move through the forestomach of a ruminant is species-specific, and can be used to classify ruminants according to their digestive physiology into 'moose-types' (with little difference in fluid and small particle passage) and 'cattle-types' (where fluids move through the forestomach much faster than small particles). So far, 'moose-types' appear limited to a dietary niche of browsing, whereas 'cattle-types' are particularly prominent in the intermediate and grazing diet niches. However, some species, including members of the spiral-horned antelopes (the Tragelaphini), have a 'cattle-type' physiology but a browse-dominated diet niche. Eland (Taurotragus oryx), the largest member of the Tragelaphini, are strict browsers in the wild but have been considered intermediate feeders in the past, and can seemingly be maintained on grass diets. We quantified food intake, mean retention time (MRT) in the gastrointestinal tract and the reticulorumen (RR) of a solute, a small and a large particle marker, and diet digestibility in six eland each fed a monocot (grass hay) and a dicot (lucerne silage) forage. Food intake and digestibility was lower on the diet with higher fibre content (grass hay), with corresponding longer MRT. At the higher intakes on lucerne, the difference in MRT between small and large particles was larger, indicating a greater reliance on particle sorting and clearance under this condition of potentially limiting gut capacity. Regardless of diet or intake, the ratio of small particle and solute MRT in the RR was constant and small, at a quotient of 1.54, classifying the eland as a typical 'moose-type' ruminant. This finding is consistent with previous literature reports on low faecal metabolic nitrogen and high apparent protein digestibility in eland. Given the relative ease at which eland can be maintained under farm husbandry conditions, they appear ideal model ruminants to study the effects of differences in rumen physiology compared to cattle.
- MeSH
- antilopy fyziologie MeSH
- bachor fyziologie MeSH
- dieta * MeSH
- dusík metabolismus MeSH
- feces MeSH
- trávení * MeSH
- velikost částic MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A 1000-cow study across four European countries was undertaken to understand to what extent ruminant microbiomes can be controlled by the host animal and to identify characteristics of the host rumen microbiome axis that determine productivity and methane emissions. A core rumen microbiome, phylogenetically linked and with a preserved hierarchical structure, was identified. A 39-member subset of the core formed hubs in co-occurrence networks linking microbiome structure to host genetics and phenotype (methane emissions, rumen and blood metabolites, and milk production efficiency). These phenotypes can be predicted from the core microbiome using machine learning algorithms. The heritable core microbes, therefore, present primary targets for rumen manipulation toward sustainable and environmentally friendly agriculture.
- MeSH
- bachor metabolismus MeSH
- fenotyp MeSH
- fylogeneze MeSH
- kohortové studie MeSH
- krev metabolismus MeSH
- methan metabolismus MeSH
- mléko metabolismus MeSH
- skot genetika mikrobiologie MeSH
- střevní mikroflóra genetika fyziologie MeSH
- zvířata MeSH
- Check Tag
- skot genetika mikrobiologie MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Gram-stain-positive, catalase and oxidase-negative and short rod-shaped bacterium C10 with occasional branching was isolated under strictly anaerobic conditions from the rumen fluid of a red deer (Cervus elaphus) in the course of study attempting to uncover new xylanolytic and cellulolytic rumen bacteria inhabiting the digestive tract of wild ruminants in the Czech Republic. The anaerobic M10 medium containing bovine rumen fluid and carboxymethylcellulose as a defined source of organic carbon was used in the process of bacterial isolation. The 16S rRNA gene similarity revealed recently characterized new species Actinomyces succiniciruminis Am4T (GenBank accession number of the gene retrieved from the complete genome: LK995506) and Actinomyces glycerinitolerans G10T (GenBank accession number from the complete genome: NZFQTT01000017) as the closest relatives (99.7 and 99.6% gene pairwise identity, respectively), followed by the Actinomyces ruminicola DSM 27982T (97.2%, in all compared fragment of 41468 pb). Due to the taxonomic affinity of the examined strain to both species A. succiniciruminis and A. glycerinitolerans, its taxonomic status towards these species was evaluated using variable regions of rpsA (length of 519 bp) and rplB (597 bp) gene sequences amplified based on specific primers designed so as to be applicable in differentiation, classification, and phylogeny of Actinomyces species/strains. Comparative analyses using rpsA and rplB showed 98.5 and 97.9% similarities of C10 to A. succiniciruminis, respectively, and 97.5 and 97.6% similarities to A. glycerinitolerans, respectively. Thus, gene identities revealed that the evaluated isolate C10 (=DSM 100236 = LMG 28777) is a little more related to the species A. succiniciruminis isolated from the rumen of a Holstein-Friesian cow than A. glycerinitolerans. Phylogenetic analyses confirmed affinity of strain C10 to both recently characterized species. Unfortunately, they did not allow the bacterial strain to be classified into a particular species. Phenotypic characterization suggested similar conclusions. This brief contribution is aimed at classification and detailed phenotypic characterization of bacterial strain C10 isolated from the rumen of a wild red deer exhibiting, from the point of view of Actinomyces species, noteworthy cellulolytic and xylanolytic activities.
- MeSH
- Actinomyces klasifikace genetika izolace a purifikace metabolismus MeSH
- bachor mikrobiologie MeSH
- bakteriální geny genetika MeSH
- celulosa metabolismus MeSH
- DNA bakterií genetika MeSH
- fenotyp MeSH
- fylogeneze MeSH
- mastné kyseliny analýza MeSH
- peptidoglykan analýza MeSH
- RNA ribozomální 16S genetika MeSH
- vysoká zvěř mikrobiologie MeSH
- xylany metabolismus MeSH
- zastoupení bazí MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In this study, the effects of seven pure plant secondary metabolites (PSMs) on rumen fermentation, methane (CH4 ) production and rumen bacterial community composition were determined. Two in vitro trials were conducted. In trial 1, nine concentrations of 8-hydroxyquinoline, α-terpineol, camphor, bornyl acetate, α-pinene, thymoquinone and thymol were incubated on separate days using in vitro 24-hr batch incubations. All compounds tested demonstrated the ability to alter rumen fermentation parameters and decrease CH4 production. However, effective concentrations differed among individual PSMs. The lowest concentrations that reduced (p < .05) CH4 production were as follows: 8 mg/L of 8-hydroxyquinoline, 120 mg/L of thymoquinone, 240 mg/L of thymol and 480 mg/L of α-terpineol, camphor, bornyl acetate and α-pinene. These concentrations were selected for use in trial 2. In trial 2, PSMs were incubated in one run. Methane was decreased (p < .05) by all PSMs at selected concentrations. However, only 8-hydroxyquinoline, bornyl acetate and thymoquinone decreased (p < .05) CH4 relative to volatile fatty acids (VFAs). Based on denaturing gradient gel electrophoresis analysis, different PSMs changed the composition of bacterial communities to different extents. As revealed by Ion Torrent sequencing, the effects of PSMs on relative abundance were most pronounced in the predominant families, especially in Lachnospiraceae, Succinivibrionaceae, Prevotellaceae, unclassified Clostridiales and Ruminococcaceae. The CH4 production was correlated negatively (-.72; p < .05) with relative abundance of Succinivibrionaceae and positively with relative abundance of Ruminococcaceae (.86; p < .05). In summary, this study identified three pure PSMs (8hydroxyquinoline, bornyl acetate and thymoquinone) with potentially promising effects on rumen CH4 production. The PSMs tested in this study demonstrated considerable impact on rumen bacterial communities even at the lowest concentrations that decreased CH4 production. The findings from this study may help to elucidate how PSMs affect rumen bacterial fermentation.
- MeSH
- bachor metabolismus mikrobiologie MeSH
- dieta veterinární MeSH
- dusík metabolismus MeSH
- fermentace * MeSH
- fyziologie bakterií MeSH
- kyseliny mastné těkavé metabolismus MeSH
- lipnicovité metabolismus MeSH
- methan biosyntéza MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
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.
- MeSH
- bachor metabolismus MeSH
- dieta MeSH
- fyziologie výživy zvířat fyziologie MeSH
- isoflavony aplikace a dávkování analýza metabolismus MeSH
- krmivo pro zvířata MeSH
- laktace MeSH
- siláž MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Dairy goats are often fed a high-concentrate (HC) diet to meet their lactation demands; however, long-term concentrate feeding is unhealthy and leads to milk yield and lactose content decreases. Therefore, we tested whether a buffering agent is able to increase the output of glucose in the liver and influence lactose synthesis. Eight lactating goats were randomly assigned to two groups: one group received a HC diet (Concentrate : Forage = 6:4, HG) and the other group received the same diet with a buffering agent added (0.2 % NaHCO(3), 0.1 % MgO, BG) over a 19-week experimental period. The total volatile fatty acids and lipopolysaccharide (LPS) declined in the rumen, which led the rumen pH to become stabile in the BG goats. The milk yield and lactose content increased. The alanine aminotransferase, aspartate transaminase, alkaline phosphatase, pro-inflammatory cytokines, LPS and lactate contents in the plasma significantly decreased, whereas the prolactin and growth hormone levels increased. The hepatic vein glucose content increased. In addition, pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6PC) expression in the liver was significantly up-regulated. In the mammary glands, the levels of glucose transporter type 1, 8, 12 as well as of sodium-glucose cotransporter 1 increased. Cumulative buffering agent treatment increased the blood concentrations of glucose via gluconeogenesis and promoted its synthesis in the liver. This treatment may contribute to the increase of the milk yield and lactose synthesis of lactating goats.
- MeSH
- bachor účinky léků metabolismus MeSH
- dieta MeSH
- glukoneogeneze účinky léků MeSH
- kozy metabolismus MeSH
- krmivo pro zvířata MeSH
- kyseliny mastné neesterifikované metabolismus MeSH
- laktace účinky léků MeSH
- laktosa metabolismus MeSH
- lipopolysacharidy metabolismus MeSH
- mléko chemie MeSH
- pufry MeSH
- růstový hormon fyziologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
This study aimed to isolate and characterize a novel cellulolytic enzyme from black goat rumen by using a culture-independent approach. A metagenomic fosmid library was constructed from black goat rumen contents and screened for a novel cellulase. The KG37 gene encoding a protein of 858 amino acid residues (92.7 kDa) was isolated. The deduced protein contained a glycosyl hydrolase family 74 (GH74) domain and showed 77% sequence identity to two endo-1,4-β-glucanases from Fibrobacter succinogenes. The novel GH74 cellulase gene was overexpressed in Escherichia coli, and its protein product was functionally characterized. The recombinant GH74 cellulase showed a broad substrate spectrum. The enzyme exhibited its optimum activity at pH 5.0 and temperature range of 20-50 °C. The enzyme was thermally stable at pH 5.0 and at a temperature of 20-40 °C. The novel GH74 cellulase can be practically exploited to convert lignocellulosic biomass to value-added products in various industrial applications in future.
- MeSH
- bachor mikrobiologie MeSH
- celulasa chemie genetika izolace a purifikace MeSH
- Escherichia coli genetika metabolismus MeSH
- exprese genu MeSH
- Fibrobacter enzymologie genetika MeSH
- genetické testování MeSH
- genová knihovna MeSH
- klonování DNA MeSH
- koncentrace vodíkových iontů MeSH
- kozy mikrobiologie MeSH
- metagenom * MeSH
- metagenomika MeSH
- molekulová hmotnost MeSH
- rekombinantní proteiny genetika izolace a purifikace metabolismus MeSH
- sekvenční homologie MeSH
- stabilita enzymů MeSH
- substrátová specifita MeSH
- teplota MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
To study the various processes in the rumen the in vitro techniques are widely used to realize more controlled and reproducible conditions compared to in vivo experiments. Mostly, only the parameters like pH changes, volatile fatty acids content or metabolite production are monitored. In this study we examine the bacterial community dynamics of rumen fluid in course of ten day cultivation realize under standard conditions described in the literature. Whereas the pH values, total VFA content and A/P ratio in bioreactor were consistent with natural conditions in the rumen, the mean redox-potential values of -251 and -243mV were much more negative. For culture-independent assessment of bacterial community composition, the Illumina MiSeq results indicated that the community contained 292 bacterial genera. In course of ten days cultivation a significant changes in the microbial community were measured when Bacteroidetes to Firmicutes ratio changed from 3.2 to 1.2 and phyla Proteobacteria and Actinobacteria represented by genus Bifidobacterium and Olsenella significantly increased. The main responsible factor of these changes seems to be very low redox potential in bioreactor together with accumulation of simple carbohydrates in milieu as a result of limited excretion of fermented feed and absence of nutrient absorbing mechanisms.
- MeSH
- bachor metabolismus mikrobiologie MeSH
- Bacteria klasifikace genetika růst a vývoj izolace a purifikace MeSH
- Bacteroidetes genetika růst a vývoj izolace a purifikace MeSH
- bioreaktory mikrobiologie MeSH
- DNA bakterií MeSH
- fermentace MeSH
- Firmicutes genetika růst a vývoj izolace a purifikace MeSH
- kyseliny mastné těkavé metabolismus MeSH
- mikrobiota * genetika MeSH
- oxidace-redukce MeSH
- Proteobacteria genetika růst a vývoj izolace a purifikace MeSH
- RNA ribozomální 16S genetika MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
High-grain feeding used in the animal production is known to affect the host rumen bacterial community, but our understanding of consequent changes in goats is limited. This study was therefore aimed to evaluate bacterial population dynamics during 20 days adaptation of 4 ruminally cannulated goats to the high-grain diet (grain: hay - ratio of 40:60). The dietary transition of goats from the forage to the high-grain-diet resulted in the significant decrease of rumen fluid pH, which was however still higher than value established for acute or subacute ruminal acidosis was not diagnosed in studied animals. DGGE analysis demonstrated distinct ruminal microbial populations in hay-fed and grain-fed animals, but the substantial animal-to-animal variation were detected. Quantitative PCR showed for grain-fed animals significantly higher number of bacteria belonging to Clostridium leptum group at 10 days after the incorporation of corn into the diet and significantly lower concentration of bacteria belonging to Actinobacteria phylum at the day 20 after dietary change. Taxonomic distribution analysed by NGS at day 20 revealed the similar prevalence of the phyla Firmicutes and Bacteroidetes in all goats, significantly higher presence of the unclassified genus of groups of Bacteroidales and Ruminococcaceae in grain-fed animals and significantly higher presence the genus Prevotella and Butyrivibrio in the forage-fed animals. The three different culture-independent methods used in this study show that high proportion of concentrate in goat diet does not induce any serious disturbance of their rumen ecosystem and indicate the good adaptive response of caprine ruminal bacteria to incorporation of corn into the diet.
- MeSH
- Actinobacteria klasifikace genetika metabolismus MeSH
- bachor mikrobiologie MeSH
- Bacteroidetes klasifikace genetika metabolismus MeSH
- Butyrivibrio klasifikace genetika metabolismus MeSH
- Clostridium klasifikace genetika metabolismus MeSH
- fermentace MeSH
- Firmicutes klasifikace genetika metabolismus MeSH
- fylogeneze MeSH
- fyziologie výživy zvířat * MeSH
- koncentrace vodíkových iontů MeSH
- kozy MeSH
- krmivo pro zvířata analýza MeSH
- kukuřice setá chemie metabolismus MeSH
- lipnicovité chemie metabolismus MeSH
- píštěl žaludku MeSH
- Prevotella klasifikace genetika metabolismus MeSH
- Ruminococcus klasifikace genetika metabolismus MeSH
- sekvenční analýza DNA MeSH
- střevní mikroflóra fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
