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two-step freezing procedure for cryopreservation of rumen ciliates, an effective tool for creation of a frozen rumen protozoa bank.the present study aimed at the long-term storage of rumen protozoa as living cells in liquid nitrogen. the two-step or interrupted slow freezing procedure was used to cryopreserve six of the dominant species of rumen ciliates isolated from monofaunated animals, dasytricha ruminantium, entodinium caudatum, epidinium ecaudatum caudatum, eudiplodinium maggii, isotricha prostoma, and polyplastron multivesiculatum. we optimized the first step in the interrupted slow freezing procedure, from the extra ...200312839750
effect of antibiotics, 2-bromoethanesulfonic acid and pyromellitic diimide on methanogenesis in rumen ciliate cultures in vitro.the effects of penicillin g, streptomycin, chloramphenicol, 2-bromoethanesulfonic acid and pyromellitic diimide on total gas, methane, volatile fatty acid production and food degradability after 24 h of incubation in vitro were investigated in the cultures of two rumen ciliates. the inocula of both rumen ciliates entodinium caudatum and epidinium ecaudatum were used at a volume of 34 ml into the 50 ml glass syringes together with the feed and compounds tested. despite penicillin g--streptomycin ...200111851015
the 3' untranslated region of messages in the rumen protozoan entodinium caudatum.the 3' untranslated regions of a number of cdnas from the rumen protozoal species entodinium caudatum were studied with a view to characterising their preference for stop codons, general length, nucleotide composition and polyadenylation signals. unlike a number of ciliates, entodinium caudatum uses uaa as a stop codon, rather than as a codon for glutamine. in addition, the 3' untranslated region of the message is generally less than 100 nucleotides in length, extremely a+t rich, and does not ap ...200010965953
analysis of intraspecific sequence variation among eight isolates of the rumen symbiont, isotricha prostoma (ciliophora), from two continents.the internally transcribed spacer regions 1 and 2 (its-1 and its-2) and the 5.8s ribosomal rna gene of isotricha prostoma were examined for intraspecific sequence variation. there were no differences in the its-1/5.8s/its-2 region among cattle and sheep isolates of i. prostoma from australia, canada, and the united states, indicating that this region is 100% conserved among eight isolates from two continents.199910461387
phylogeny of the rumen ciliates entodinium, epidinium and polyplastron (litostomatea:entodiniomorphida) inferred from small subunit ribosomal rna sequences.there complete 18s ribosomal rna gene sequences from the rumen ciliates, entodinium caudatum (1,639 bp), epidinium caudatum (1,638 bp), and polyplastron multivesiculatum (1,640 bp) were determined and confirmed in the opposite direction. trees produced using maximum parsimony and distance-matrix methods (least squares and neighbour-joining), with strong bootstrap support, depict the rumen ciliates as a monophyletic group. entodinium caudatum is the earliest branching rumen ciliate. however, ento ...19979172834
assessment of ciliates in the sheep rumen by dgge.this work was carried out to develop a rapid molecular profiling technique to screen ciliate populations in the rumen of sheep.200415242452
the cryopreservation of some large ciliate entodiniomorphid protozoa taken from the rumen.large entodiniomorphid ciliates taken from the rumen cannot be reliably kept in vivo, and cryopreservation was investigated as an alternative approach to storage. three species of large entodiniomorphid ciliated (polyplastron multivesiculatum, epidinium ecaudatum forma caudatum and ophryoscolex caudatus forma tricoronatus) were cryopreserved by one-step freezing, with mean recoveries of 43, 51 and 80%, respectively. for ophryoscolex caudatus forma tricoronatus two-step freezing was investigated. ...19968987899
comparative study of rumen ciliates in buffalo, cattle and sheep in egypt.rumen ciliates species and composition of the sheep, friesian-cattle and water buffaloes in egypt were surveyed. as a result, 7 genera with 18 species and 6 formae in sheep, 10 genera with 28 species and 11 formae in cattle and 12 genera with 29 species and 7 formae in water buffaloes were detected. twenty-two species were common in both cattle and buffaloes, while 12 species in sheep were common with the other ruminants. entodinium spp., such as e. simplex, e. nanellum and e. exigum, appeared m ...19968877984
polypeptides of hydrogenosome-enriched fractions from rumen ciliate protozoa and trichomonads: immunological studies.the evolution of hydrogenosomes, energy-generating organelles of rumen ciliate protozoa and the flagellate trichomonads has been the subject of much speculation. polypeptides of the hydrogenosome-enriched fractions from the rumen ciliates, dasytricha ruminantium, isostricha spp., polyplastron multivesiculatum and eudiplodinium maggii were separated by sds-page and compared to analogous polypeptide preparations from tritrichomonas foetus. immunoblotting with antisera specific to the hydrogenosome ...19948181726
the effects of organic selenium supplementation on the rumen ciliate population in sheep.the effect of selenium supplementation on the rumen protozoan population of sheep was demonstrated. both the total and generic counts of rumen ciliates in sheep fed a diet with basal se content (70 microg/kg dry matter) were compared to those of animals given feed supplemented with inorganic (disodium selenite) or organic se (selenized yeast) (310 microg/kg dry matter). the genera of entodinium, isotricha, dasytricha, ophryoscolex, diploplastron and polyplastron occurred in all sheep except for ...200516408856
some rumen ciliates have endosymbiotic methanogens.most of the small ciliate protozoa, including dasytricha ruminantium and entodinium spp. living in the rumen of sheep, were found to have intracellular bacteria. these bacteria were not present in digestive vacuoles. they showed characteristic coenzyme f420 autofluorescence and they were detected with a rhodamine-labelled archaea-specific oligonucleotide probe. the measured volume percent of autofluorescing bacteria (1%) was close to the total volume of intracellular bacteria estimated from tem ...19948181718
digestion of a dehydrated lucerne/barley diet (70:30) in defaunated, isotricha-monoinoculated and mixed-fauna-inoculated rumen in sheep.two adult texel sheep fitted with permanent rumen fistulae were defaunated for 12 weeks in a preliminary experiment. they were then reinoculated with isotricha sp for 11 weeks, and finally with a mixed fauna (entodinium, epidinium, eudiplodinium and isotricha) for 10 weeks. they were fed a diet composed of dehydrated lucerne (700 g/d), pelleted barley grain (300 g/d), grass hay (100 g/d) and wheat straw (50 g/d) in one daily meal. isotricha and the ciliates of the mixed fauna developed rapidly, ...19957612165
the importance of methanogens associated with ciliate protozoa in ruminal methane production in vitro.the importance of methanogenic bacteria associated with ciliate protozoa was estimated either by removing protozoa from whole rumen fluid (using defaunated rumen fluid to correct for the effects of centrifugation on bacteria) or by isolating the protozoa. rumen fluid was withdrawn from sheep inoculated with either polyplastron multivesiculatum, a co-culture of isotricha prostoma plus entodinium spp. or a mixed type b fauna of entodinium, eudiplodinium and epidinium spp. methanogenesis was highes ...19957576513
[effect of rumen ciliates on the digestion of different carbohydrates in sheep. i.--utilization of cell wall carbohydrates (cellulose and hemicellulose) and of starch].two diets rich in cell-wall carbohydrates or starch were given to 10 rumen-fistulated sheep; two sheep were defaunated and the others were inoculated with polyplastron multivesiculatum (p) or entodinium sp. (e), or both (p + e), or with conventional fauna. ciliate biomass was greater when the animals were fed a high starch diet than when the diet was rich in cell-wall carbohydrates (table 2). with both diets, the entodinium genus in the mixed fauna sampling predominated. we showed that polyplast ...19826818644
[effect of cobalt and vitamin b 12 on growth and survival of rumen ciliates in vitro as a function of bacterial population].the effect of inorganic cobalt or organic cobalt in the form of vitamin b12 was studied on the growth of rumen ciliates in the presence of normal bacterial population and in the presence of reduced bacterial population. it was stated that concentration from 0.1 to 1 cobalt microgram/ml and 0.12 to 0.24 vitamin b12 microgram/ml stimulated growth of rumen ciliates. at rates 10 to 20 times weaker, cobalt in the form of vitamin b12 was more active than cobalt in inorganic form. with reduced bacteria ...19826818641
[attempt to obtain and maintain rumen entodiniomorph ciliates in axenic cultures].a method to obtain and maintain rumen ciliates in either axenic cultures or cultures with a reduced flora is described; it consists of two or three incubating periods with different antibiotic associations (separated by washing) and renewing of nutrient medium. on these conditions, polyplastron multivesiculatum is obtained in an axenic state and can survive for 5 days. entodinium sp. can survive for 10 to 14 days, but with a reduced bacterial population composed only of strict aerobic of faculta ...19826816119
association of methanogenic bacteria with ovine rumen ciliates.1. the frequency of association between methanogenic bacteria and ovine rumen ciliates was studied in the rumen fluid of a fistulated sheep. 2. a period of fasting and flushing of the rumen content with nitrogen resulted in a relatively high association, whereas the intake of food and flushing with hydrogen caused a detachment of the methanogenic bacteria from the ciliates. 3. the changes in the frequency of association can be correlated with the relative attribution to the h2 production by hydr ...19826800402
a descriptive study of rumen digestion in meroxenic lambs according to the nature and complexity of the microflora.we studied in meroxenic lambs, i.e. in lambs with a simplified digestive microflora, the effect of the microflora on the quantities of solid feed intake and on the main digestive parameters in the rumen. axenic lambs were inoculated with a more or less complex flora, obtained by diluting (10(-8), 10(-7), 10(-8) a pool of rumen fluid taken either from young conventional lambs before weaning from adult sheep (pool a) or from meroxenic lambs (pool b). a few of these lambs then were inoculated with ...19836648032
[effect of rumen ciliates on the digestive utilization of various carbohydrate-rich diets and on the end-products formed in the rumen. ii. utilization of inulin, saccharose and lactose].three diets rich in inulin, saccharose and lactose, respectively, were given to 10 rumen-fistulated sheep. two animals were defaunated, two were inoculated with either polyplastron multivesibulatum or entodinium sp., and two others were inoculated with both. the latter two were bred in conventional conditions. all animals ingested the same amounts of carbohydrates in the three diets (21-22 g/kg p0.75/day). dietary nitrogen content was similar (table 1). the ciliate population was improved with t ...19836412334
rumen ciliate protozoa: effects on digestion in the stomach of sheep.six wethers, each fitted with a rumen cannula and duodenal reentrant cannula, were used to study effects of ciliate protozoa on rumen digestion and metabolism. a corn: corn silage (1:1) diet was fed for two periods. during the first period, defaunation was attempted with nonyl phenol ethylene oxide. defaunation was complete in three sheep and partial in the other three sheep in which a reduced population of a small entodinium sp. was observed. during the second period the sheep were inoculated w ...19836308075
establishment of ciliate protozoa in the rumen of conventional and conventionalized lambs: influence of diet and management conditions.the establishment of ciliate protozoa in the rumen was studied in conventional lambs reared under different conditions of management. the role of the microflora in the kinetics of this establishment was also investigated in conventionalized lambs. in lambs reared under farm conditions ciliate protozoa appeared in the following order: entodinium (15-20 days), polyplastron, eudiplodinium, and epidinium (20-25 days), and isotricha (50 days). entodinium was the most abundant (10(5)-10(6) ciliates ml ...19883138013
the role of ciliate protozoa in nutrition of the ruminant.the effects of ciliate protozoa on the ruminal ecosystem, digestion in different parts of the gut, the nature of nutrients available for absorption and their effects upon the nutrition and productivity of their host are reviewed. compared with fauna-free ruminants, the presence of ciliate protozoa results in a more stable ruminal fermentation, higher levels of ammonia, reduced numbers of bacteria, as well as changes in dry matter (%), liquid volume and turnover rate of ruminal contents. associat ...19863098727
composition of the rumen ciliate population in experimental herds of cattle and sheep in lethbridge, alberta, western canada.rumen ciliate populations were surveyed in 11 holstein cattle and 6 sheep in lethbridge (alta., canada) to determine species distribution in this western canadian environment. a total of 28 ciliate species were identified in cattle and 17 in sheep. the average total number of ciliates per millilitre of rumen content was 6.9 x 10(4) in cattle and 1.9 x 10(5) in sheep. the average number of species per host was 20.5 in cattle and 13.8 in sheep. of the ciliate species detected, species of entodiniu ...19892504472
enzymatic properties of pyruvate kinase from the rumen ciliates genus entodinium.1. pyruvate kinase from the rumen ciliates genus entodinium was partially purified and the enzymatic properties were investigated. 2. three types of pyruvate kinase (type i, ii and iii) on deae-cellulose column were eluted with a linear gradient of kcl. the enzymatic properties differed among the types of enzyme, especially type i and type iii displayed different kinetic properties to each other. the enzymatic property of type ii enzyme was an intermediate between type i and iii. 3. the principa ...19892503406
experimental reproduction of phytobezoars.in a series of in virtro experiments (n = 64) using the mature flowers or seeds and pappus hairs of karoo bushes chrysocoma ciliata, dimorphotheca cuneata, eriocephalus ericoides, gazania krebsiana and gnidia polycephala, small phytobezoars were formed from material of e. ericoides, g. krebsiana and g. polycephala either alone or in combinations. the ratio of fibre to liquid used was found to be optimal around 1: 7 and milling of material enhanced bezoar formation. time and ph did not appear to ...19921328637
physical structure and chemical composition of abomasal phytobezoars of goats and sheep.the diameters of fibres from phytobezoars varied between 2 and 30 microns, with an average 58% measuring less than < 15 microns, 36% between 15 and 23 microns, and 6% greater than 23 microns. these diameters were very similar to those of the pappus hairs surrounding the seeds of the karoo bushes chrysocoma ciliata, eriocephalus ericoides and gnidia polycephala (means of 6.06 microns, 16.67 microns and 22.73 microns respectively). the microscopic structure and ultrastructure of these pappus hairs ...19921328636
phylogenetic study of methanogens associated with rumen ciliates.the phylogeny of methanogenic archaea associated with ciliate protozoa in a sheep rumen was investigated. ruminal ciliate protozoa were exhaustively washed and mixtures of genomic dna extracted. archaea-specific nested pcr amplification was conducted with the ciliate genomic mixture. the resultant small subunit (16s) ribosomal rna gene (ssu rdna) was cloned into escherichia coli jm 109. many methanogens were still observed on and/or in ciliate cells by fluorescent microscopy even after exhaustiv ...199910441724
a re-appraisal of the diversity of the methanogens associated with the rumen ciliates.the diversity of methanogenic archaea associated with different species of ciliated protozoa in the rumen was analysed. partial fragments of archaeal ssu rrna genes were amplified from dna isolated from single cells from the rumen protozoal species metadinium medium, entodinium furca, ophryoscolex caudatus and diplodinium dentatum. sequence analysis of these fragments indicated that although all of the new isolates clustered with sequences previously described for methanogens, there was a differ ...200415358415
effect of nisin on two cultures of rumen ciliates.the effect of nisin (in the form of nisaplin) was determined using two species of rumen ciliate protozoa in vitro, on their co-culture bacterial population, and volatile fatty acid concentration. nisaplin did not affect the in vitro growth of entodinium caudatum at concentrations of 50-400 mg/l during short-term treatment (5 d). long-term application (30 d) of nisaplin (100 mg/l) significantly decreased growth of the epidinium ecaudatum forma caudatum et ecaudatum but not growth of e. caudatum. ...200312879756
potential of galvanotaxis to separation and cleaning of rumen ciliates.the ability of rumen ciliate protozoa to move in a unidirectional electrical field from the anode to the cathode was tested in large-volume electromigration equipment; the aim was to concentrate the microorganisms and clean them of impurities. during galvanotaxis in the freshly harvested rumen fluid and at a voltage of 10 v (i=0.8 ma), cells of isotricha (isotricha prostoma, isotricha intestinalis) were the first to swim towards the cathode; 1 min later, they were followed by dasytricha ruminant ...200415003689
xylanases and carboxymethylcellulases of the rumen protozoa polyplastron multivesiculatum, eudiplodinium maggii and entodinium sp.endoglucanase and xylanase activities of three rumen protozoa, polyplastron multivesiculatum, eudiplodinium maggii, and entodinium sp. were compared qualitatively by zymograms and quantitatively by measuring specific activities against different polysaccharides. a set of carboxymethylcellulases and xylanases was produced by the large ciliates whereas no band of activity was observed for entodinium sp. in zymograms. specific activity of endoglucanases from p. multivesiculatum (1.3 micromol mg pro ...200515727834
comparison of nisin and monensin effects on ciliate and selected bacterial populations in artificial rumen.the effect of daily supplementation of nisin (2 mg/l), monensin (5.88 mg/l) and nisin and monensin (2 + 5.88 mg/l) on ovine ruminal ciliates and bacteria was investigated using the artificial rumen rusitec. major groups in rusitec were entodinium spp. and dasytricha ruminantium. the supplementation of nisin significantly increased the population of both major ciliate groups. the supplementation of monensin significantly decreased the population of both groups. the combined effect of nisin and mo ...201020140721
bacterial-protozoal interactions in a microbial community of rumen ciliate entodinium caudatum culture under mercury stress.we examined the role of rumen ciliates, using entodinium caudatum as a model organism, in the detoxification of soluble mercury(ii) in vitro under conditions with enhanced or reduced diversity of a co-culture bacterial population as well as the effects of long-term mercury(ii) stress on in vitro fermentation parameters and major mercury detoxification products. the e. caudatum growth depended on the capability of the co-culture bacterial population to develop resistance to mercury(ii) chloride a ...201020453906
occurrence of hydrogenosomes in the rumen ciliates ophryoscolecidae. 198214719520
why does the establishment of the starch preferring entodinium caudatum in the rumen decrease the numbers of the fibrolytic ciliate eudiplodinium maggii?the effect of the establishment of entodinium caudatum on the population of eudiplodinium maggii was examined in the rumen of three sheep fed a hay/ground barley diet. the cell concentration of e. maggii were 15.9-38.5 and 11.7-12.4 x 10(3) cells per g of the rumen contents in the absence and presence of e. caudatum, respectively. microscopic analysis showed that starch was the only material engulfed by eudiplodinia irrespective of the time after feeding and the presence or absence of e. caudatu ...200415227784
regeneration of cryoresistance of in vitro rumen ciliate cultures.the purpose of this study was to investigate factors affecting mechanical- and cryo-resistance of the rumen ciliates entodinium caudatum (e.c.), entodinium furca monolobum (e.f.m.), entodinium simplex (e.s.), diplodinium denticulatum (two clones, d.d.01 and d.d.02), diploplastron affine (d.a.) and epidinium ecaudatum forma caudatum (e.e.c.) after long-term in vitro cultivation. following prolonged in vitro cultivation (more than six months), the ciliates were very sensitive to both centrifugatio ...200515950963
chitinolytic activity of the sheep rumen ciliate diploplastron affine.supplementation of the rumen ciliate diploplastron affine growth medium with commercial chitin stimulated growth of ciliates and the density of their population was positively correlated with chitin doses (r = 0.95; p < 0.01). the cell-free extracts prepared from bacteria-free ciliates degraded chitin to n-acetyl-d: -glucosamine and chitobiose. three exochitinases, two endochitinases and two beta-n-acetylglucosaminidases were identified in the cell-free extract of protozoa. the molar mass of exo ...200818661291
mureinolytic ability of the rumen ciliate diploplastron affine.rumen ciliate protozoa intensively engulf bacteria. however, their ability to utilize murein which is the main polysaccharide of bacterial cell wall has hardly been recognized. the present study concerns the ability of the rumen protozoa diploplastron affine to digest and ferment murein. the ciliates were isolated from the rumen fluid and grown in vitro or inoculated into the rumen of defaunated sheep. the results of long-term cultivation of protozoa showed a positive correlation between their n ...201020680560
the ability of the rumen protozoan eudiplodinium maggii to utilize chitin.the ability was determined of the rumen ciliate eudiplodinium maggii to utilize chitin from fungal cell wall. cultivation experiments shoved that the population concentration (number of ciliates in vitro) was positively correlated with chitin doses. cell extract prepared from the bacteria-free ciliates degraded colloidal chitin releasing 2.0 micromol reducing sugar per mg protein per h. end products of this reaction were chitotriose and n-acetylglucosamine. incubation of the bacteria-free ciliat ...201020680569
characterization of rumen ciliate community composition in domestic sheep, deer, and cattle, feeding on varying diets, by means of pcr-dgge and clone libraries.the structure and variability of ciliate protozoal communities in the rumens of domestic new zealand ruminants feeding on different diets was investigated. the relative abundance of ciliates compared with bacteria was similar across all samples. however, molecular fingerprinting of communities showed ruminant-specific differences in species composition. community compositions of cattle were significantly influenced by diet. in contrast, diet effects in deer and sheep were weaker than the animal- ...201121204869
phylogeny of trichostome ciliates (ciliophora, litostomatea) endosymbiotic in the yakut horse (equus caballus).ciliates of the subclass trichostomatia inhabit the fermentative regions of the digestive tract of herbivores. most available small subunit ribosomal rna (ssrrna) gene sequences of trichostomes are from species isolated from the rumen of cattle or sheep and from marsupials. no ciliate species endosymbiotic in horses has yet been analyzed. we have sequenced the ssrrna genes of five ciliate species, isolated from the cecum and colon of four yakut horses: cycloposthium edentatum, cycloposthium ishi ...200717720462
suitability of different media for in vitro cultivation of the ruminal protozoa species entodinium caudatum, eudiplodinium maggii, and epidinium ecaudatum.three protozoal cultivation media were tested to determine the medium which best facilitated growth and viability of key b-type ciliates isolated from the sheep rumen. entodinium caudatum and eudiplodinium maggii were grown anaerobically in 50-ml flasks for 32 days in caudatum-type (c), kisidayova (k) or dehority (m) medium. on day 32, in media k and m, e. caudatum cell counts were high with 5.6×10(3) and 7.8×10(3)ml(-1), respectively, and the proportion of dead cells was low with 0.6 and 1.4%, ...201121641778
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