The Cultivation of the Rumen Ciliate Entodinium simplex

Coleman, G.

doi:10.1099/00221287-57-1-81

Cited by 20 publications

(16 citation statements)

References 11 publications

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“…As a result of these observations stock cultures were maintained thereafter without rumen fluid. Epidinium ecaudatum caudatum therefore differs from both the entodinia studied previously (Coleman, 1960(Coleman, , 1969 where the presence of autoclaved rumen fluid increased the population density almost ten times. The effect of saIt concentration on the growth of Epidinium ecaudaturn caudatuni from a small inoculum in the presence of 10 :( prepared, fresh, rumen fluid.…”

Section: Effect Of Medium Constituentsmentioning

confidence: 82%

“…Although entodinia from the rumen have been successfully cultivated for long periods by several workers including Hungate ( I 943), Coleman (I 960), Tompkin, Purser & Weiser (1966), Jarvis & Hungate (1968) and Coleman (1969), there have been fewer investigations on the growth of the larger rumen entodiniomorphid protozoa. Hungate (1942Hungate ( , 1943) maintained several diplodinia, including Diplodinium (Polyplastron ?)…”

Section: Introductionmentioning

confidence: 99%

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The Cultivation of the Rumen Ciliates Epidinium ecaudatum caudatum and Polyplastron multivesiculatum in vitro

Coleman

Davies

Cash

1972

Journal of General Microbiology

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SUMMARYPopulations of over 400 Epidiniirm ecaudatum caudatumlml were maintained for over I 8 months in the presence of bacteria, in a potassium phosphate-rich medium under 95 % N,+ 5 % CO, with the daily addition of wholemeal flour and dried grass. Polyplastron multivesiculatum was also maintained at a population density of 45 to 70/ml under the same conditions, except that it was necessary to add each day a culture of another protozoon such as Epidinium ecaudatum caudatum, E. ecaudatum tricaudatum or Eudiplodinium maggii to give a population density of approximately roo/ml. The epidinia at least were engulfed by the polyplastron up to 10 being taken up each day; the minimum daily requirement for growth was one epidinium/polyplastron. Polyplastron multivesiculatum also grew in the absence of other protozoa on a sodium chloride-rich medium, and if these protozoa were inoculated into the potassium phosphate-rich medium in the presence of epidinia, there was a lag of 3 to 5 days before they began to engulf the epidinia. The effect of other variations in the culture conditions is also described. The present paper describes the cultivation of Epidinium ecaudatum caudatum and Polyplastron inultivesiculatum in the presence of bacteria in buffered salts medium with wholemeal flour and dried grass as the sources of carbohydrate. Under certain conditions P. multivesiculatum had an absolute requirement for the epidinia for growth and died rapidly without them.Preliminary communications gave the growth requirements of these ciliates immediately after their isolation from the rumen (Coleman, 1970(Coleman, , 1971).

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Section: Effect Of Medium Constituentsmentioning

confidence: 82%

Section: Introductionmentioning

confidence: 99%

The Cultivation of the Rumen Ciliates Epidinium ecaudatum caudatum and Polyplastron multivesiculatum in vitro

Coleman

Davies

Cash

1972

Journal of General Microbiology

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“…Even using the method described above, individual protozoa taken from the rumen frequently failed to grow. As with other ciliate protozoa (Coleman, 1960(Coleman, , 1969a, the population densities of the cellulolytic protozoa were always smaller in vitro than in vivo.…”

Section: Discussionmentioning

confidence: 99%

“…All experiments were made in 50 ml centrifuge tubes approximately threequarters filled with salts medium to which was added L-cysteine hydrochloride (neutralized immediately before use; final concentration 0.02 %, w/v) and, sometimes, 10 % (v/v) prepared fresh rumen fluid (Coleman, 1958) or autoclaved rumen fluid (Coleman, 1960).…”

Section: I45mentioning

confidence: 99%

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The Cultivation of Cellulolytic Protozoa Isolated from the Rumen

Coleman

Laurie

Bailey

et al. 1976

Journal of General Microbiology

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S U M M A R YConditions are described for the isolation from the rumen and subsequent growth of six species of cellulolytic protozoa : Enoploplastron triloricatum, Eudiplodinium maggii, Diploplastron afine, Epidinium ecaudatum caudatum, Diplodinium monacanthum and Diplodinium pentacanthum. The protozoa were grown in an atmosphere of 95 % nitrogen plus 5 % carbon dioxide, or pure carbon dioxide, in a potassium phosphate-rich medium containing cysteine, sometimes I o prepared fresh rumen fluid, and daily additions of powdered dried grass. Population densities of 10 to 6000 protozoa/ml (depending on the species) were obtained in cultures that were diluted with fresh medium twice each week. Extracts of these protozoa digested a [14C]cellulose preparation, liberating soluble 14C-labelled compounds. I N T R O D U C T I O NOne of the most characteristic reactions in the rumen is the breakdown and fermentation of cellulose (Tappeiner, 1884) and ciliate protozoa have long been implicated in the process (Trier, 1926). However, there have been few studies on the growth and metabolism of rumen cellulolytic protozoa. Hungate (I 942, I 943) cultured Eudiplodinium neglecturn, Eu. maggii, Diplodinium multivesiculatum and Diplod. denticulatum in the presence of ground dried grass and demonstrated that extracts of these protozoa produced reducing sugars from cellulose.We have experienced difficulty in repeating Hungate's work, and so made a fresh study of the conditions required for isolation and maintenance of six species of entodiniomorphid protozoa from the rumen, and for their growth on ground dried grass as sole additional source of organic material, apart from contaminating bacteria.As some protozoa, e.g. Entodinium spp. (Coleman, 1960(Coleman, , 1969a, have an absolute requirement for starch grains for growth, even in the presence of ground dried grass, it has been assumed that they cannot obtain sufficient energy for growth by fermentation of cellulose. Thus, any protozoon that can grow on dried grass alone can probably degrade cellulose. Initially this criterion was used to determine if an isolated protozoon was cellulolytic, and subsequently the result was confirmed by the action of extracts on pure cellulose. M E T H O D SBasal mineral salts. Caudatum-type salts medium contained (g/Ioo ml) : K,HPO,, 0.63 ;

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