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A Study of Methane-Oxidizing and Methanol-Oxidizing Bacteria." (1964)

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A Study of Methane-Oxidizing and Methanol-Oxidizing Bacteria. Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1964 A Study of Methane-Oxidizing and Methanol- Oxidizing Bacteria. Peter Konrad Stocks Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Stocks, Peter Konrad, "A Study of Methane-Oxidizing and Methanol-Oxidizing Bacteria." (1964). LSU Historical Dissertations and Theses. 1001. https://digitalcommons.lsu.edu/gradschool_disstheses/1001 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. This dissertation has been 65—3400 microfilmed exactly as received STOCKS, Peter Konrad, 1927- A STUDY OF METHANE- AND METHANOL- OXIDIZING BACTERIA. Louisiana State University, Ph.D., 1964 Bacteriology University Microfilms, Inc., Ann Arbor, Michigan A STUDY OF METHANE- AND METHANOL-OXIDIZING BACTERIA A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Department of Bacteriology by Peter Konrad Stocks B.S., Millsaps College, 1959 M .S., University of Southern Mississippi, 1961 „ . August, 1964 PLEASE NOTE: Figure pages are not original copy. They tend to ’’curl". Filmed in the best possible way. University Microfilms, Inc. ACKNOWLEDG MENT The writer wishes to express his sincere gratitude to Dr. C. S. McCleskey for his suggestions and criticism during this investigation and in the preparation of the dissertation. He gratefully acknowledges the help and understanding extended to him by Drs. A. R. Colmer, A. D. Larson, M .D. Socolofsky, A. F. Novak and J. A. Liuzzo. He especially wishes to thank Dr. A. R. Colmer for his help in the preparation of the photographic material. The writer also owes a great debt of gratitude to his friends and fellow graduate students for their inspiration, aid and criticism during the course of this study. He also wishes to thank the Office of Naval Research for the financial support of this investigation. This dissertation is dedicated to my family whose love and encouragement made this work possible. ii TABLE OF CONTENTS Page I ACKNOWLEDGMENT. ........................................................ IT- II LIST OF TABLES ..................................................................... vi III LIST OF FIGURES........................................................................ viii IV ABSTRACT.................................... x. V INTRODUCTION........................................................................... 1 VI REVIEW OF LITERATURE............................................................ 3 VII MATERIALS AND METHODS...................................................... 17 A. The Methane-Oxidizing Bacteria.................................... 17 C u ltu re s ............................................................................. 17 M e d ia................................................................................ 20 Isolation procedures.................................................... 25 Source of samples ..................................................... 25 Culture apparatus ..................................................... 25 Isolation technique............................. 29 Purity tests.................... 31 Inhibitory agents as aids in the isolation of methane oxidizers............................................... 31 Morphological studies.................................................. 33 S t a i n s.......................................................................... 33 Photomicrography..................................................... 33 Physiological and nutritional studies..................... 33 iii Page Carbon sources........................................................... 34 Nitrogen sources........................................................ 34 Serological studies.................................................... 35 B. The Pink-Pigmented Methanol-Oxidizing Bacteria. 36 C u ltu re s ............................................................................... 36 M e d ia.................................................................................. 36 Morphological studies..................................................... 41 S t a i n s............................................................................ 41 Assay for poly-B-hydroxybutyric ........................ac id 41 Physiological and biochemical studies..................... 42 Serological studies........................................................... 46 VIII RESULTS AND DISCUSSION........................................................ 47 A. The M ethane-Oxidizing B acteria ........................ 47 Enrichment and isolation of the methane oxidizers . 47 Studies with inhibitory agents................................ 49 Isolation by agglutination......................................... 55 Discussion of enrichment and isolation procedures. 57 Morphological characteristics...................................... 59 Cultural characteristics .................................................. 61 Physiological characteristics...................................... 63 The effect of methanol on the growth of the methane oxidizers.................................................... 68 iv Page The effect of NaCl on the growth of the --------------------- methane oxidizers.......................... 70 The effect of various dyes on the growth of the methane oxidizers............................................... 75 Serological relationships............................................... 77 Discussion of results obtained with methane - oxidizing bacteria.............................................................. 77 B. The Pink-Pigmented Methanol Oxidizers........................ 80 Isolation................................................................................ 80 Characteristics of pink-pigmented methanol- o x id iz e r s............................................................................. 83 Morphological characteristics................................. 83 Cultural characteristics ............................................ 83 Biochemical and physiological characteristics . 88 Studies with antibiotics.................................................. 95 Serological studies.................................................... 98 Discussion of the pink-pigmented methanol- oxidizing bacteria.............................................................. 101 IX SUMMARY.......................................................................................... 105 X LITERATURE C IT E D ........................................................................ 108 XI VITA..................................................................................................... 113 v LIST OF TABLES Page 1 S ources of-methanegQxldlzlna cultures ..................... 18 2 Sources of organisms used in comparative studies . 19 3 The composition of additional mineral salts solutions employed in this investigation.............................................. 22 4 Sources of pink-pigmented methanol oxidizers............... 37 5 Pink-pigmented methanol oxidizers obtained from other investigators............ ............................... 38 6 The effect of various antibiotics on the growth of the methane oxidizers............................................................. 51 7 The effect of various antibiotics in suppressing the growth of contaminants in methane enrichments 54 8 Utilization of hydrocarbons as sole carbon source by methane-oxidizing bacteria..........................., ........................ 65 9 Substrates tested for their ability to support the growth of methane-oxidizing bacteria................................ 67 10 The availability of various nitrogen compounds for growth and methane consumption by methane o x id iz e r s....................... 69 11 The effect of methanol on the growth of the methane- oxidizing bacteria............................ ................... 71 12 The effect of sodium chloride on the growth of the methane oxidizers in Brown's medium................................ 73 13 The effect of various dyes on the growth 'of a methane oxidizer (strain PBG)........................................ 76 14 Serological relationships among methane oxidizers and morphologically related organism............................. s 78 vi TABLE Page 15 Biochemical reactions of pink-pigmented methanol oxidizers . ......... 90 16 Growth of the pink-pigmented methanol oxidizers on various carbon substrates........................................... 93 17 Availability of various nitrogen sources for growth of methanol oxidizers.................................................................... 96 18 The effect of various antibiotics on the growth of methanol oxidizers......................................................................... 97 19 Serological relationships among pink-pigmented methanol oxidizers........................................................................ 100 vii LIST OF FIGURES FIGURE Page 1 Diagram of the apparatus used for measuring and mixing the g a se s........................................................................
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