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Environmental Biology for Engineers and Scientists D.A. Vaccari, P.F. Strom, and J.E. Alleman © John Wiley & Sons, 2005 Chapter 10 – Microbial Groups Figure 10-1. Portrait of Antonie van Leeuwenhoek Figure 10-2. Some of Leeuwenhoek's "Animalcules" from the "Scurf of the Teeth" - Drawings of Bacterial Shapes. A and B appear to show rods, with C and D showing the movement of B; E shows cocci; F rods or filaments; and G a spiral. From Leeuwenhoek' s letter of 1683 Figure 10-3. Leeuwenhoek's Microscope Figure 10-4. Portrait of Louis Pasteur Figure 10-5. Pasteur's Swan-Neck Biological Flasks Figure 10-6. A 3-Dimensional Surface: Visualizing prokaryotic species as more stable (valleys), and hence more likely, combinations of characteristics, although intermediates may exist Figure 10-7. Example of a Simplified Dichotomous Key for Identifying Filamentous Organisms in Activated Sludge Figure 10-8. Rod-Shaped Bacteria: Pseudomonas Figure 10-9. Cocci: Staphylococcus Figure 10-10. Spiral-Shaped Bacteria: Rhodospirillum Figure 10-11. A Filamentous Bacteria Growing with Floc in an Activated Sludge Wastewater Treatment Plant 10-12. Stalked Bacteria Growing on a Filament in Activated Sludge Bacterial Cell Stalk Figure 10-13. The Gram Stain Technique. This modification (one of many) is recommended for staining of activated sludge mixed liquor Figure 10-14. Bacterial Cell Wall and Membrane: a). Gram Positive; b). Gram Negative Figure 10-15. Staining Approaches + Basic (+) dyes penetrate to cytoplasm Acid (-) dyes usually do not penetrate membrane without modification (e.g., esterification) A B Differential staining with contrasting dyes to provide visual separation within cell (e.g., Gram) Cell wall staining to darken and define cell wall Spore staining used to identify cytoplasmic endospores (e.g., malachite green, with heating) Inclusion staining used to visualize cellular inclusions (e.g., polyphosphate) Flagella staining to enhance visual image of flagella Capsule staining to visually emphasize exocellular layer of encapsulating polysaccharide material Negative staining to create darkened background which offers better visual contrast for cell or capsule Figure 10-16. The Biolog Test; 95 test compounds and a control well are included in each plate. The plate shown was used to identify a Gram negative bacteria as Leminorella grimontii based on comparing the pattern of positive (dark) and negative tests to results in a database Figure 10-17. Fatty Acid Methyl Ester (FAME) profiles showing different patterns for a) Serratia marcescens and b) Tsukamurella paurometabolum a. b. Figure 10-18. Denaturing Gradient Gel Extraction (DGGE) Track Profiles Figure 10-19. Phylogenetic Tree Indicating Evolutionary Branching and Distance between Groups based on rRNA Analysis. Fungi are represented by Coprinus (a mushroom), plants by Zea (corn), and Animals by Homo (humans) Figure 10-20. Anabaena, a Filamentous Cyanobacteria; with Heterocyst Heterocyst Figure 10-21. Light Absorption Characteristics of Phototrophic Bacteria Figure 10-22. Two Species of Beggiatoa in Samples from RBC Wastewater Treatment Plants; a Gliding Filamentous Sulfur Oxidizing Proteobacteria. Note internally deposited sulfur granules Figure 10-23. Sphaerotilus natans: a) pure culture showing sheath and PHB granules; and b) branching filament in activated sludge sample a b Figure 10-24. Characteristic Zoogloea ramigera floc from activated sludge Figure 10-25. Escherichia coli Escherichia coli, Transmission Electron Micrograph Transmission Electron Image of Escherichia Eubacteria (Source: Revised from original TEM image photographed at the Central Microscopy Research and Learning Facility, University of Iowa, 85 EMRB Iowa City, IA 52242, Web Site: http://lime.weeg.uiowa.edu/~cemrf/index.html) Figure 10-26. Desulfovibrio. Note the bent rods (vibrios). Desulfovibrio sp. Sulfur-Reducing Eubacteria Figure 10-27. Endospores in Bacillus Bacillus sp. with Internal Spores (Source: pg. 1021, R.M. Atlas, Principles of Microbiology, 2nd Edition, W.C. Brown Publishers, 1997) Bacillus sp. Eubacteria with Internal Spore (~34,000x TEM Image) (Source: pg. 1021, R.M. Atlas, Principles of Microbiology, 2nd Edition, W.C. Brown Publishers, 1997) Figure 10-28. Nocardia-like Filamentous Bacteria in Activated Sludge Foam (Gram stained preparation). Figure 10-29. Spirochetes in Activated Sludge Figure 10-30. Giardia Figure 10-31. Bodo in Activated Sludge. Figure 10-32. Amoeba in Activated Sludge. Figure 10-33. Arcella, a Testate Amoeba Figure 10-34. Free Swimming Ciliates: a) Paramecium, b) Aspidisca, and c) Euplotes. Figure 10-35. Vorticella, a Stalked Ciliate: a) feeding; b) with mouth closed, and myoneme visible (dark line in stalk); c) stalk extended, and d) seconds later, myoneme contracted to form corkscrew-shaped stalk. a b c d Figure 10-36. Podophyra, a Suctorean Figure 10-37. Cryptosporidium Figure 10-38. Euglena Euglena sp. Euglenoid Algae (Source: B. Leander, Center for Ultrastructural Research, University of Georgia, Athens, Georgia, Web Site: http://www.uga.edu/~caur/home.html) Figure 10-39. SEM Images of Various Diatom Frustules. SEM Images of Various Diatom Shell Structures (Source: Central Microscopy Research and Learning Facility, University of Iowa, 85 EMRB Iowa City, IA 52242, Web Site: http://lime.weeg.uiowa.edu/~cemrf/index.html) Figure 10-40. Peridinium Peridinium Dinoflagellate Algae Ceratium sp. Dinoflagellate (Source: pg. 550, L.M. Prescott, J.P. Harley, and D.A. Klein, Microbiology, 4th Edition, WCB/McGraw-Hill, 1999) Chlorella sp. Green Algae Figure 10-41. Scenedesmus Figure 13-30 Scenedesmus sp. Green Algae Figure 10-42. Mold with Conidia Mold with Budding Condidia Tip Structures (Source: Central Microscopy Research and Learning Facility, University of Iowa, 85 EMRB Iowa City, IA 52242, Web Site: http://lime.weeg.uiowa.edu/~cemrf/index.html) Figure 10-43. Nematode trapping fungus. Figure 10-44. The Combined Effect of Mycorrhyzal Fungus and Phosphate Fertilizer on Tomato Growth. g dry weight of leaf 1 0.8 0.6 w/o fungus w/fungus 0.4 0.2 0 0.6 1.2 mEq phosphate per plant Figure 10-45. Virus Capsids