135 Report - Isolation and Characterization of Microbes...

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Isolation and Characterization of Microbes Alex Katler Partner: Erica Wagner Lab. Section: 2 Report For Experiments: 1,3,5
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Introduction and Results: The goal of this experiment was to isolate two different microbes form samples of different natural environments. All procedures, such as plating or any other type of inoculations, were done using aseptic technique. The first sample was taken from a southern Wisconsin out-door soil sample. The weather of this region is indicative of the types of conditions this sample was exposed to. Wisconsin has hot and humid summer months, sometimes reaching the high 90’s Fahrenheit, and sub-zero winter months (which is when the sample was taken). It was also subjected to moderate precipitation year-round. The second sample was a green pea taken from a sealed leftover container vegetable soup. The soup had been refrigerated (35-40 o F) for a long duration of time, possibly on the order of weeks. Initially, .1g of each sample was used to inoculate a 125ml flask of liquid medium. All-purpose NGY liquid was inoculated with a pea from the soup sample and minimal NEM liquid was inoculated with the soil sample. Each flask was then incubated on a shaker for 5 days at 30 o C. Slight turbidity was observed in the NGY flask after this incubation period and not in the NEM flaks. Gram stains of both cultures revealed multiple types of microbes. The NEM (soil) gram stain revealed a majority of spread out gram-positive rod shaped microbes and several types of coccus gram-positive ones as well. The NGY (pea) gram stain showed more diversity. There were gram-positive and negative microbes of many different shapes. A wet mount of the NEM medium revealed some flagellated organisms. The wet mount of the NGY liquid showed that some of the rods were connected end to end. Plates of homologous medium plus agar were then streaked with the microbe cultures and incubated at 37 o C. Two days of incubation allowed for enough growth for the re-streaking of single colonies. The NEM colonies were small, flat, white, circular and surrounded by matted dark fuzzy looking matter that rose several millimeters above the surface of the medium. The NGY colonies were
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off-white, flat and circular. After two re-streak and incubation intervals, the gram stains showed that gram-positive rod shaped microbes had been isolated for both cultures. Both cultures were then re- streaked onto NGY medium. The soil colony morphology changed to larger, off-white, raised and circular while the pea cultures’ colonies stayed the same. Both cultures were then transferred to NGY slant tubes and incubated at 30 o C. From this point on these tubes were used as a source of inoculum. To characterize nutritional and oxidative relationships in microbes, several tests were performed
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135 Report - Isolation and Characterization of Microbes...

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