BURROUGHS, ANI) WILLIAM C. CHRISTIANSEN. Continuous culture of ruminal microorganisms in chemically defined medium. II. Culture medium studies. Appl. Microbiol. 10:583-592. 1962. Ruminal ciliates have been grown in continuous culture in chenmically defined media and in the absence of viable bacteria. Oligotrichic ruminal ciliates seem to reqjuire insoluble carbohydrates for growth; the

QUINN, LOYD Y. (Iowa State University, Ames). Continuous culture of ruminal microorganisms in chemically defined medium. I. Design of continuous-culture apparatus. Appl. Microbiol. 10:580-582. 1962.-An apparatus is described which has been used for successful continuous culture of the ciliates from the rumen of cattle. Automatic control of feeding rate, pH, oxidation-reduction potential, temperature, stirring rate, aeration rate, salinity, and volume of the culture is provided for, using standard commercial equipment, whenever possible. The operation of this apparatus is described.

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Results and discussion 169 / y Influence of Diethylstilbestrol Upon Rumen Protozoal Numbers and the Effect of Protozoa Upon Growth Performance of Lambs l80 Methods and procedures l8l Results and discussion 186 CONCLUSIONS AND SUMMARY 192 GENERAL DISCUSSION 195 LITERATURE CITED 200 ACKNOWLEDGEMENTS 211 APPENDIX 212 8 strated with glucose as an end product of cellulose breakdown. Hungate (1943) showed that rumen protozoal breakdown of cellu lose led to a rapid synthesis of food reserves, such as amylopectin or other starch granules. Further studies by Hungate revealed a rapid storage and utilization of starch by rumen protozoa. On the other hand, Heald et al. (1952) indicated that maltose and lactose were not utilized, by rumen protozoa. Oxford (1955) postulated that many of the carbohydrates, such as sucrose, glucose, cellobiose, insulin and levans, were used by protozoa to synthesize amylopectln. Sugden (1953) observed that excess soluble carbohydrates, such as glucose and sucrose, caused protozoal cells to burst. Since four-fifths of the cell weight may be amylopectln, additional soluble carbohydrates may cause additional reactions leading to excess carbon dioxide gas which probably is the cause of cell bursting, especially in Diplodinium (Sugden, 1953). Studies by Heald and Oxford (1953) indicated that Isotrlchs and Dasytricha produce high levels of rumen glucose, fructose, sucrose, raffinose, inulin, rye grass levan and cellobiose. The formed acids were primarily acetic, butyric and lactic, with traces of propionic. Utilization of many of the soluble sugars by rumen proto zoa has been demonstrated by several investigators (Sugden, 1953; Oxford, 195 lj Hungate, 1942 and 1943; Gutierrez, 1953; Heald _et _al., 1952). Hungate ( 1942) showed that holotrichs readily utilized glucose, fructose, sucrose, galactose, cello biose, inulin and levan. Sugden (1953) and Hungate (1943)

Methods and procedures 20 Results and discussion.. . • 26 In Vitro Volatile Fatty Acid Production by Rumen Protozoa ' 29 Responses of rumen protozoa to pH changes and time of in vitro fermentations 31 Methods and procedures 31 Results and discussion 37 Responses of rumen protozoa to substrate and protozoal concentration changes of in vitro fermentations 41 Methods and procedures 42 Results and discussion 44 Responses of rumen protozoa bo diethylstilbestrol and antibiotics of in vitro fermenta tions. 48 Methods and procedures 49 Results and discussion 51 Responses of rumen protozoa to sodium, potas sium and phosphorus of in vitro fermentations 57 Methods and procedures 58 Results and discussion 63 CONCLUSIONS AND SUMMARY ' 78 ill Page PART II. PRODUCTION OF VOLATILE FATTY ACIDS AND AMMONIA IN THE RUMEN OF FAUNATED •AND PROTOZOA-FREE LAMBS 82a REVIEW OF LITERATURE 82b Techniques for Obtaining Protozoal-Free Lambs.. .. 82b In Vivo Production-of Rumen Volatile Fatty Acids and Ammonia in Faunated and Protozoal-Free Lambs. 89 EXPERIMENTAL PROCEDURE AND RESULTS 98 Rearing Isolated NewBorn Orphan Lambs 98 Methods and procedures ' 98 Results and discussion 105 Def'aunating the Rumen of Lambs Having Protozoal Populations Methods and results Discussion Volatile Fatty Acid Production in the Rumen of Faunated, Partially Faunated and Protozoal-Free Lambs Methods and procedures.. Results and discussion.. .. Ruminai Ammonia Production in the Rumen of Faunated, Partially Faunated and Protozoal-Free Lambs.. .. 138 Methods and procedures 139 Results and discussion.. .