Streak plate for Environmental Sampling. 4 Quadrant Streak Plate 1. swab 2. loop 3. loop 4. loop.

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Presentation transcript:

Streak plate for Environmental Sampling

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

Four Quadrant Streak Plate

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

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

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

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

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.

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

spores thallus Fungal Classification: molds

Fungal classification: yeasts

Budding KM

Dimorphism

Penicillium notatum

Penicillium 100x 400x KM

Penicillium spores

Rhizopus 100x KM

Rhizopus

Aspergillus 100x 400x KM

Candidiasis Cutaneous fungal infections

Tinea pedis Dermis.net, used with permission

Tinea corporis Dermis.net, used with permission

Tinea cruris, ringworm of the groin Cutaneous fungal infections

Candidiasis

Sporotrichosis DermAtlas, used with permission

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

A few last points Systemic mycoses True Fungal infections Opportunistic infections

The life cycle of Aspergillus Spores inhaled Germination Mass of hyphae (plateau phase) Hyphal elongation and branching & Fungal Research Trust, used with permission

Aspergillosis in the brain WebPath, used with permission

Mucomycosis eye infection CDC, used with permission

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

Staining

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

Mechanism of Direct Stains Add a positively charged blue dye

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

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

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

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

Examining Indirect Stains KM

Example of a Negative Stain KM

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

What do you observe? What do you conclude?

Always look carefully Used with permission

Always look carefully KM

Bacterial Shapes KM

Bacterial Arrangements KM

Gram reactions KM Used with permission

Staining Bacteria

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