Bio 163 Post-lab PDF
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Jay-Ann A. Tallad
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Summary
This document details three exercises on microbial ecology focusing on air, water, and soil samples. It describes methods for collecting and characterizing microbes, including procedures like gravitational settling, hay infusion, and contact slide assay. Microbes such as protozoa, cyanobacteria, and microalgae are highlighted. The document also explains methods for microbial analysis and identification.
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BIO 163 POST LAB Jay-Ann A. Tallad Exercises 01 Microbial Ecology of Air 02 Microbial Ecology of Water 03 Microbial Ecology of Soil Exercise 1: Microbial Ecology of Air OBJECTIVES 1. To collect bioaerosol samples using gravitational settling 2. To morphologically characterize microbial co...
BIO 163 POST LAB Jay-Ann A. Tallad Exercises 01 Microbial Ecology of Air 02 Microbial Ecology of Water 03 Microbial Ecology of Soil Exercise 1: Microbial Ecology of Air OBJECTIVES 1. To collect bioaerosol samples using gravitational settling 2. To morphologically characterize microbial colonies Gravitational settling Settling plate - media plate for the growth of microorganisms direct method of checking the air for viable microorganisms exposed to main areas of activity such that they capture maximum amount of particles large particles tend to settle faster due to gravitational force smaller particles tend to settle slower due to air currents driving them away Colony Characteristics STANDARD PLATE COUNT - used to quantify microbial load Crowded Plate: TNTC Normal Plate: 30 - 300 CFUs TFTC Spreader Plate Spreader plates Chains of cells not too distinctly separated that appears to be caused by the disintegration of bacterial clumps. Spreader develops in the film of water between the agar and the bottom of the plate Spreader develops in the film of water at the edge or on the surface of the agar. Exercise 2: Microbial Ecology of Water Objective 1. to isolate and identify protozoa, cyanobacteria, and microalgae from freshwater sources Hay Infusion Sugars in dried grass provide food for bacteria and other microbes The bacteria serve as food for the protozoa If the bacteria grow quickly, the protozoa will also grow quickly. Hay Infusion Unpigmented, moving microorganisms are usually protozoans (Click me) Guide to identification of protozoans, algae and cyanobacteria Greenish or golden-brown organisms are usually algae. Organisms that appear bluegreen will be cyanobacteria Protozoa - produce cysts, resistant dormant stage, that allows them to survive drought, heat, and freezing Phylum Sarcomastigophora Subphylum Sarcodina (Amoebae): move via pseudopodia, has amoeboid movement Subphylum Mastigophora (Zooflagellates): move via flagella Phylum Ciliophora: movement is via cilia Phylum Apicomplexa: lack locomotor organelles and all are internal parasites Algae- includes all photosynthetic eukaryotic organisms in Kingdom Protista - have visible nuclei and chloroplasts that contain chlorophyll a Cyanobacteria- blue-green algae - photosynthetic, utilizing chlorophyll a Exercise 3: Microbial Ecology of Soil (Click Me) Contact Slide Assay and Microscopy Objective 01 1. To understand the spatial distribution and relationship of soil microorganisms Objective 02 2. To identify the effect of abiotic factors to these soil microorganisms Contact Slide Assay buried-slide technique useful for observing soil fungi, actinomycetes, and bacteria via microscopy HOAc: fixes the microorganisms on the glass slide STAIN: Lactophenol Cotton Blue Phenol Kills any live microorganism Lactic acid Preserves fungal structures Cotton Blue Stains chitin in fungal cell wall microorganisms tend to reside in the water located within soil pores bacteria (small, rounded shapes due to nutritional stress --> more favorable surface: volume ratio) actinomycetes (thin strings) fungal hyphae (thick threads) treated soil will have more soil microbes urea: provide nitrogen source glucose: provide carbon source spatial interactions can be visualized soil particles are dark, irregular shapes fungi have thick filamentous hyphae actinomycetes have thin filamentous hyphae bacteria are found as small cocci or rod shapes, typically in clumps, on soil particles or lining fungal hyphae ALSO REVIEW THE GUIDE QUESTIONS IN YOUR LAB REPORTS BIO 163 1ST LAB EXAM NOVEMBER 22, 2023 (WEDNESDAY) KA 201 1:00 - 3:00