Eastern Illinois University The Keep Masters Theses Student Theses & Publications 1979 Siderocapsa Major-- Fact or Artifact? Ray W. Swanson Eastern Illinois University This research is a product of the graduate program in Zoology at Eastern Illinois University. Find out more about the program. Recommended Citation Swanson, Ray W., "Siderocapsa Major-- Fact or Artifact?" (1979). Masters Theses. 3120. https://thekeep.eiu.edu/theses/3120 This is brought to you for free and open access by the Student Theses & Publications at The Keep. It has been accepted for inclusion in Masters Theses by an authorized administrator of The Keep. For more information, please contact [email protected]. 1 ACliliOWLEDGFl·IENTS The author would to his most sincere appreciation like express to Dr. William Weiler for his u.�crring and encouragement through­ A. guidance out this study. Particular thanks chould go to Dr. William Jame::; for his also s. :\dvice and help in the scanning el tron a.nd X-ray ec microsco["J analysis 't<3chniques, as reading of well as the r.ianuscrlpt.. In addition, indebtedness is expressed to Dr. HacLeod, Director R. and Dr. R. McConnville, Assistant Director of the Center for Electron Hicroscopy (University of Illinois, Champaign-Urban.a) and m m rs of their e be staff for the scanning electron rnicrcscopy and X-ray assistance in o.na!y�is tachnlqucs Speer (Botany De�-rt:nent, E.I.U.) �c to Dr. John f'vr mce.. 1n ..:.:t13ht ass.!.!;t nic�osc·)py tec!mlqucs. Spacial t!1anks go to ar:d Dr. Max Dr. WilUam T. Rid3eway Chapnan thci:r help in the and for constructive c:·lticism a.l'ld. prepar<tt,ion writing he is. of this t s Finally, ut or wizhes th:::.r.k sis er, the a h to Je;:.�1fe�, his t for her Gaintly and typi:sg patience while read.ir:g the ;rianuscrip-t.. 11 TA B!$ OF CONTENTS Page ACKNOWLEDGMENTS • • • • • • • • • 1 ABSTRACT • • • • • • • • • • 111 LIST OF FIGURES AND TABLES • • • • v INTRODUCTION • • 1 HISTORICAL • • • • • Types of Iron Bacteria • • • • • • • 3 • • • • • Taxonomy of the Heterotrophic Iron Bacteria 6 Physiology • • • • • • • ' . 17 Autrophy vs. Heterotrophy • .. • • . - . 1? • • • • • • • • Cultural Investigations . 22 Cultural l-1ethods. • • • • • • • • • • • • • • 22 Cultural Requirements • • • • • • • • • • • 2.3 Carbon Sources • • • • • • • • • • 23 Nitrogen Sources • . • ,: • • • • • ... • • • • • • 25 Mineral Sources. • • • • • • • • • • • 2? Growth Factors • • • • • • • .ll • • • 28 pH • • • • • • • • • • • • • • • • • 29 'I'empera ture. • • • • • • • • • • • • • • • • • • • • 29 Salt Tolerance • • • • • • • • • • • • 29 . Storage Products • • • • • • • • • • • • 30 • Ecology and Economic Importance • • • • • • • • • • • 31 MEl'HODS AND MATERIAIS • • • • • • • • • • 34 General • • • • • • • • • • • • • • 34 Light nicroscopy. • • • • • • • • • • • • • • 35 Scanning Electron Microscopy. • • • . • • • J6 X-ray Microanalysis • • • • • • • J6 Cultural Techniques • • • • • • • • • • • • • • • • • • 37 RESULTS AND DISCUSSION. • • • • • • • • • • 38 CONCWSIOi�S. • • • •· . • • 60 61 APPENDIX A. • • • • • • LITERATURE CITED. • • . • • • 63 iii ABSTRACT Swanson, Ray W. M.S., Ea.stem Illinois University. August 1979. Siderocapsa major-- Fact or Artifact? Major Professors Dr. William A. Weiler. The validity of the genus Siderocapsa, a group of unicellular iron- depositing bacteria, has been a subject of debate for many years, Ia.ck of photomicroscopic and cultural studies has kept this group in taxonomic obscurity. Bacteria were collected from the Campus pond, Eastern Illinois University, Charleston, Illinois, by a submerged -. slide/coverslip technique, 'Ihese.bacteria were identified as .Siderocapsa major according to the description of Molisch (1909). Scanning electron micros�opy of Siderocapsa major microcolonies has shown cells (0.5 by 1.15 )im), in a clear central well (2-6 ).llll), surrounded by an amorphous material which ac�umulates with the age of the colony. Most authors have failed to explain how they have concluded that the material �--·_: sequestered by Siderocapsa microcolonies is iron and/or manganese. Prelimtnary work employing the Prussian blue R�action (and modifications) failed to demonstrate the presence of iron in the capsules around these bacteria. X-ray microanalysis also failed to demonstrate the presence of iron. However significant amounts of manganese were found to be present in the extra-cellular material. Some investigators have suggested that Siderocapsa microcolonies are artifacts and the result of _rod-shaped bacteria that have become entrapped in the well of a Sphaerotilus natans holdfast whose filamen� iv has become dislodged. Scanning electron microscopy has shown that the size and morphology of the holdfast of Sphaerotilus-�ptothrix members does not correspond to the size of the deposit around the Siderocapsa microcapsule. In addition, statistical analysis has shown a definite developnental pattern relating the total number of cells in the micro­ capsule to the total amount of material deposited. The author concludes, from material observed here, that the Siderocapsa microcolonies are discretely:·different entities from members of the Sphaerotilus-leptothrix group but may be related to other heterotrophic bacteria. , v LIST OF FIGURES AND TAB� Figure Page 1. Light micrograph of a typical Siderocapsa microcolony • • • • . 40 2. Light micrograph of a Siderocapsa microcolony showing . , , ·· • • 40 cells inside the: central·. well. J. Scanning electron micrograph of a young Siderocapsa major microcolony • , • • • • • • • � • • • • • • • , • • • • • • • 40 4. Scanning electron' micrograph of a more mature Siderocapsa · major microcolony • , • • • • • • • • • • • • • • • • • • • • 40 4 5, Light micrograph of �haerotilus natans • • • • • • • • • • • 3 6. Light micrograph of Sphaerotilus natans cells stained with • • • • • • • • • • • • • • • • • • • • • • • • 4) Sudan Black B . t micrograph a filamentous member of the 7. Scanning elec ron of 4J Sphaerotilus-I.eptothrix group • • • • • • • • • • • • • • • • 8. Scanning electron micrograph of several members of the Sphaerotilus-1.eptothrix group showing differences in sheath 4 and hold.fast morphology • • • • • • • • • • • ! • • • • • • • J 9. Light micrograph of Sphaerotilus natans showing Prussian blue 4 Reaction • • • • • • • • • • • • • • • • • • • • • • • • • • J .10. Light micrograph of Sphaerotilus natans showing Prussian blue stai?ing of the sheath only • • • • • • • • • • • • • • • • • 4J 11. X-ray analysis of a young Sphaerotilus nata.ns filament • • • 46 12. X-ray analysis of the area�:9f ·deposition of a Siderocapsa .•, microcolony • • • • • • • • • • . • • • • • • • • • • • • • • • 46 ·' - I . • 1J. Scanning electron micrograph of a young Siderocapsa mnjor microcolony • • • • • • • • • • • • • • • • • • • • • • • • • 50 14. Scanning electron micrograph of a maturing Siderocapsa major microcolony • • • • • • • • • • • • • • • • , • • • • • • • • 50 15. Scanning electron micrograph of mo.derately developed and fully developed Siderocapsa major microcolonies • • • , • • • 50 16. Scanning electron micrograph of Siderocapsa major showing cellular reproduction • • • • • • • • • • • • • • • • • • • • 50 vi AND LIST OF FIGURES TABLES (Continued) Figure Page 17. Scanning electron micrograph of Sphaerotilus natans • • • • 52 18. Light micrograph of Siderocapsa showing filaments emerging from the area of depositi.on • • • • • • • · , • • • • • • • • 52 , 19. Scanning electron micrograph of SiderocaP§a ma.ior shoving a fi�ent emerging from the area of deposition •••••• 52 Scanning electron micrograph of Siderocapsa major showing 20. 52 a filament arising from the area of deposition ••••••• 21. Light micrograph of a Siderocapsa microcolony showing Sphaerotilus natans attached to the area of deposition • • • 55 . 22. Light micrograph of cells of the iron-oxidizing isolate from· a .. 72-hour PCA culture showing V-shaped arrangments typical of Arthrobacter • • • • •. • • • • • • • • • • • • • • ·55 23. Light micrograph of cells of the iron-oxidizing bacterium showing sheath fonnation due to the presence of calcium • • • 55 / 24. Light micrograph of cells from a 4 week old culture of the .:.... iron-oxidizing bacterium • • • • • • • • • • • • • • • • • • 55 Tables 1. Microcolony morphology of Siderocapsa major • • • • • • • • • 48 2. Physiological test results· • • • • • • • • • • • • • • • • • 59 1 rnTRODUCTION Iron is present in natural waters in solution and in particulate floes, '!be universal occurrence of iron deposits in water system pipes and on the surfaces of leaves· of aquatic plants suggests the presence of bacteria which have the ability to sequester ir�n. 'lbe iron bacteria;: axe;;c�paole 1 -0f•-depost.tirfg/fer�f:c! or··ma:ngarli:c oxides without regard to the physiological significance of this function to the bacteria themselves. Because of this the term iron bacteria . refers not to a taxonomic group_but to an ecological group, This is exemplified by the diversity in habitat seen among the members of this group. Chemoautotrophic species exist in aoid mine waters. Hetero­ troph�c iron bacteria are found in neutral to alkaline waters. These two types have essentaally nothing in common except the ability to · accumulate iron and/or manganese, Much confusion has evolved from attempts to taxonomically 1dent1fy the heterotrophic members, specifically the Sphaerotilus-I.eptothrix grpup and the Siderocapsaceae.· Ambiguous results and conflicting . reports from cultural studies have not reduced the confusion. The validity