1. Streak plate for Environmental Sampling

2. 4 Quadrant Streak Plate 1. swab 2. loop 3. loop 4. loop

3. Four Quadrant Streak Plate

4. Red algae site:oceanexplorer.noaa.gov, used with permission

5. Agar Properties Melts 100 C but liquid till 40 C Not toxic Not “degraded” Lacks nutrients

6. Proper Labeling Name Date Name microbe/source of inoculum Ex # Miscellaneous info

7. AsepticAseptic Technique vs. Universal Precautions Why??? How???? When???? Microbelibrary ASM, used with permission

8. To Do Take 2 TSA plates and two broths Properly label them Wet swab with sterile water…sample body with 1 swab….streak quadrant 1 of TSA plate Take swab and place in TSB tube Repeat for “environment” plate.

9. 4 Quadrant Streak Plate 1. swab 2. loop 3. loop 4. loop

11. spores thallus Fungal Classification: molds

12. Fungal classification: yeasts

13. Budding KM

14. Dimorphism

15. Penicillium notatum

16. Penicillium 100x 400x KM

17. Penicillium spores

18. Rhizopus 100x KM

19. Rhizopus

20. Aspergillus 100x 400x KM

21. Candidiasis Cutaneous fungal infections

22. Tinea pedis Dermis.net, used with permission

23. Tinea corporis Dermis.net, used with permission

24. Tinea cruris, ringworm of the groin Cutaneous fungal infections

25. Candidiasis www.meddean.luc.edu

26. Sporotrichosis DermAtlas, used with permission

27. Subcutaneous fungal infections Sporotrichosis CDC Dr. Lucille K. Georg, used with permission

28. A few last points Systemic mycoses True Fungal infections Opportunistic infections

29. The life cycle of Aspergillus Spores inhaled Germination Mass of hyphae (plateau phase) Hyphal elongation and branching http://www.aspergillus.org.uk/index.html & Fungal Research Trust, used with permission

30. Aspergillosis in the brain WebPath, used with permission

31. Mucomycosis eye infection CDC, used with permission

32. To Do Part II Prepare Fungal Cultures Glass slide agar Fungal smear

34. Staining

35. Principles of Staining Stain or dye: A molecule that can bind to a cellular structure and give it color (contrast) 1. Positively charged or Basic Dyes 2. Negatively charged or Acidic Dyes Simple stain: makes use of a single dye and reveals basic cell shapes and arrangements Differential stain: makes use of two or more dyes and distinguishes between organisms based on structural differences Wiley

36. Mechanism of Direct Stains Add a positively charged blue dye

37. Reasons for Heat Fixation  Kills the Cells  Adherence  Coagulation  Prevents Autolysis

38. Procedures SE BMEC Right NOW: Make 2 identical slides. Label 1 Gm and the other Dt 1.Smear (both slides) 2.Air dry (both slides) 3.Heat fix (both slides) Gm KM

39. Mechanism of Negative stainingNegative Field of view Microbe Add a negatively charged blue dye Animation courtesy of ASM MicrobeLibrary, used with permission

40. Watch Demo on how to make a smear for a indirect stain

41. Examining Indirect Stains KM

42. Example of a Negative Stain KM

43. 1-2 seconds Gram StainGram Stain ASM MicrobeLibrary, used with permission

44. What do you observe? What do you conclude?

45. Always look carefully Used with permission

46. Always look carefully KM

47. Bacterial Shapes KM

48. Bacterial Arrangements KM

49. Gram reactions KM Used with permission

50. Staining Bacteria

51. 1-2 seconds Gram StainGram Stain ASM MicrobeLibrary, used with permission