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Microbe 3c Lab Report Procedure

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Analysis of Major Unknown 3C Summary: A series of tests was done with Microbe 3C. A Gram stain was done and the sample was gram positive. A streak plate was completed revealing that the microbe was clear/white in color, margin was entire, shape was circular, elevation was convex/raised, and the opacity was translucent. The last test done was the sucrose test. It starts orange and turns yellow positive for microbe 3C Klebsiella oxytoca (Professor Brady, Personal Communication). Procedure 1 Gram stain: Purpose/ Introduction: The Gram stain reveals whether a microbe is gram positive or gram negative. It also reveals the shape and arrangement of the microbe (Professor Brady, Personal Communication). Of the forty-three possible microbes the following are gram positive: Bacillus cereus, Bacillus circulans, Bacillus coagulans, Bacillus megaterium, Bacillus subtilis, …show more content…

The following was eliminated because they are gram positive: Bacillus cereus, Bacillus circulans, Bacillus coagulans, Bacillus megaterium, Bacillus subtilis, Bacillus thuringiensis, Brevibacillus brevis, Geobacillus stereothermophilus, Lysinibacillus sphaericus, Clostridium perfringens, Corynebacterium pseudodiphtheriticum, Listeria monocytogenes, Lactobacillus casei, Enterococcus faecalis (Streptococcus), Streptococcus lactis (Lactococcus), Streptococcus mutans, Streptococcus pneumonia, Micrococcus (Kocuria) roseus, Sarcina lutea (Micrococcus luteus), Sporosarcina ureae, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, and Rhodococcus rhodochorous. The remaining microbes are gram negative: Alcaligenes faecalis, Rhizobium radiobacter, Salmonella typi, Serratia marcescens, Shigella flexneri, Yersina pseudotuberculosis, Erwina persicinus, Serratia liquifacens, Halobacter salinarium, Enterobacter aerogenes, Eschirichia coli, Flavobacter capsulatum,

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According to the series of test that my group ran for our unknown specimen, we had a match with the bacteria known as Alcaligenes Faecalis. This bacterium belongs to one of the major group of gram-negative bacteria (Phylum Proteobacteria). Alcaligenes Faecalis (Genus, species) is a rod shaped (bacillus), 0.5-1.2 x 1.0-3.0 µm, round with scalloped margin (colony configuration growth), motile (with one to nine peritrichous flagella), gram-negative, non-fermentative bacteria, obligate aerobic, having oxygen as the principal terminal electron acceptor in the electron transport chain (ETC). We consider we have a match with the species Alcaligenes Faecalis because of the following reasons: Fermentation tests performed (Durham sugars) were negative, which indicate that our bacteria use a different metabolic means for growth (non-fermentative gram-negative bacteria).

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The second ½ of the organism was used for gram staining. The gram stain method was performed on the unknown organism per lab manual page 42 and two gram stain reactions were identified. Organism B was gram positive cocci in grape like clusters. Because organism B was positive I could eliminate Escherichia coli, Enterobacter aerogenes and Proteus vulgaris because these bacteria would be rod shapes. Organism A was gram negative pink rod shaped and because of gram positive morphology I could eliminate Staphylococcus aureus, Streptococcus lactis and Bacillus subtilis.

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The unknown bacteria was then tested on multiple selective and differential media. Growth was present on the MacConkey Agar and the colonies were the same color as the plate, which told me my bacteria was gram negative and did not ferment lactose. There was no growth on the Mannitol Salt Agar, and this told me the unknown was not salt tolerant and did not

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After experiment on microscope under oil immersion, I learned that my Unknown is gram positive. Under the lens, the bacteria appears in purple color. Its morphology is cocci arranged in cluster. However, during decolorizing process, I put too much alcohol to the crystal violet-iodine complex making the color overly removed. That led to the result of my gram positive has slightly redish

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After a gram stain was done unknown #257 was identified as a gram positive organism because when observed under the microscope the organism appeared purple with cocci in clusters. The organism was also catalase positive which means that it produced enzyme catalase and bubbled when hydrogen peroxide was added to it. Three test were conducted based on the result of the gram staining procedure. Blood agar with a Novobiocin disk was chosen as well as DNase (DNA) and Mannitol Salts (MSA) agar. The Blood agar is a bright red, opaque plate and the streaking or the inoculation technique was a modified streaking for isolation with a heavy quadrant one.

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