Some Cultural Studies with Holotrich Ciliate Protozoa of the Sheep's Rumen

Self Cite

SUMMARY: By direct viable counts in deep cultures in a basal nutrient agar, consisting of buffered and very highly clarified nunen liquor, supplemented only with vitamin-free acid hydrolysate of casein, tryptophan, a fermentable carbohydrate and agar, the crude wet nunen contents of a hay-fed sheep has been shown to contain a population of c. lo8 viable and facultatively anaerobic bacteria per g., mostly Gram-positive cocci capable of fermenting'a wide range of soluble carbohydrates including starch and inulin but not mannitol. "he mannitol-fermenting population is of the order of lo6 organisms per g., consisting mostly of Gramnegative rods, including coliform bacteria. Addition of reducing substances such as sodium sulphide or memaptoacetate (thioglycollate) to the basal agar had little or no effect in increasing the counts. An extract of hay made With cleared and sterile rumen liquor did not yield the typical nunen streptococcal flora when counted by this method in rafFinose+rumen liquor nutrient agar.Although most workers with rumen micro-organisms seem to have assumed either that the important fermentative bacteria and protozoa in this very mixed culture must be obligate anaerobes, or that they will not act except under powerful reducing conditions (see, e.g. Hungate (1950) ; Huhtanen, Rogers & Gall (1952) and previous papers by Gall and associates on the methods for isolating nunen bacteria; Sijpesteijn & Elsden (1952) on methods of using washed suspensions of mixed rumen bacteria for fermentation studies), nevertheless there are good reasons for believing that the rumen normally contains a considerable population of facultative anaerobes which are active in fermenting carbohydrates. Thus, by a simple enrichment technique not carried on for more than 18 hr., Heald (1952a, b) isolated numerous pure cultures of coliform bacteria, capable rapidly of fermenting glucose and xylose, from the rumens of hay-fed sheep. MacPherson (1953), in a study of nunen amylolytic bacteria, without any difficulty isolated numerous facultatively anaerobic streptococci, which resembled Strep. bovis in fermentation reactions, from the rumens of both concentrate-fed and hay-fed sheep. These bacteria were absent from the fodder. Sugden & Oxford (1952) showed that an important and ubiquitous group of sheep's rumen ciliate protozoa, the holotrichs, which are very active in fermenting glucose (Heald & Oxford, 1958), are not a t all exacting in their demands for anaerobic conditions and will

Section: Methodsmentioning

confidence: 99%

“…These bacteria were absent from the fodder. Sugden & Oxford (1952) showed that an important and ubiquitous group of sheep's rumen ciliate protozoa, the holotrichs, which are very active in fermenting glucose (Heald & Oxford, 1958), are not a t all exacting in their demands for anaerobic conditions and will P . J .…”

mentioning

confidence: 99%

A Method for Direct Viable Counts of the Facultatively Anaerobic Microflora in the Rumen of a Sheep maintained on a Hay Diet

Heald¹,

Krogh²,

Mann³

et al. 1953

Self Cite

“…Clarke (1965) suggested that this decline was due to bursting of the ciliates when overfull of synthesized polysaccharide (Sugden & Oxford, 1952). Use can be made of an expression analogous to that used in the calculation of growth rate, i.e.…”

Section: A C I Warnermentioning

confidence: 99%

Diurnal Changes in the Concentrations of Micro-organisms in the Rumens of Sheep Fed Limited Diets Once Daily: WITH AN APPENDIX ON THE KINETICS OF RUMEN MICROBES AND FLOW

Warner¹

1966

SUMMARYThe pattern of change of concentration of different groups of microorganisms in the rumen was found to be characteristic of the group and little affected by the time of day, the nature of the diet, or the host animal. The dilution rate of rumen liquor and the rate of change of concentration of several groups of micro-organisms were measured a t intervals following feeding; this allowed calculation of the apparent specific growth rate. The changes in concentration of the ophryoscolecid ciliate protozoa, the selenomonads and the oval organism described by Eadie could be explained as the resultant of the changes in dilution rate due to the act of eating and in growth rate in response to incoming nutrients. Rates of change in concentration greater than could be accounted for on the basis of growth and dilution alone were found with the peptostreptococci, the polymastigate flagellate protozoa and the holotrich ciliate protozoa. It is suggested that the peptostreptococci underwent lysis or engulfment by other organisms and that the polymastigates became sequestered, .probably close to the rumen wall. The concentration changes of the holotrichs were more difficult t o understand, but it would appear that little division took place for some 18 hr after feeding, followed by several divisions in quick succession. The oscillospirae showed two peaks in concentration; no explanation can be offered for this.

“…In early attempts a t culture, the protozoa were incubated with antibiotics (K benzylpenicillin 0.004 yo, neomycin sulphate, 0.003 yo), rumen liquor, rice starch and casein, and, for the first 24 hr., with mannose, 0.5 yo, to kill the holotrichs (Sugden & Oxford, 1952). The oligotrichs in these cultures did not survive more than 5 days.…”

Section: Preliminary Investigationsmentioning

confidence: 94%

The Cultivation of Some Rumen Oligotrich Protozoa

Clarke

1963

SUMMARYThe rumen oligotrich protozoa, Eremoplastron bovis (Dogiel, 1927), E. brevispinurn (Kofoid & MacLennan, 1932), Entodifiium longifiucleatum (Dogiel, 1925), E. ovimum (Dogiel, 1927), Eudiplodinium nzaggii (Fiorentini, 1889) and various Epidinium species have been cultured in mixed culture for over 7 months. The medium consisted of rice starch, dried grass and casein in a mineral salt solution +rumen liquor. No antibiotics were used.Preliminary studies showed that sodium sulphide causes inhibition of protozoal metabolism in concentrations greater than 0.01 yo. Pure cultures of Epidinium ecaudatum (Fiorentini, 1889) and Erernoplastron bovis have been established. In addition, a clone of E . bovis was maintained for 5 months.A method for growing quantities of protozoa for biochemical studies is described.