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Bio 139 Lab Practical 2
Photos from lab exercises covered on the second lab practical exam
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E. coli on MacConkey's agar

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Compare growth on nutrient rich GYE aerobically vs anaerobically.

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Compare growth in air or anaerobically.

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Compare growth aerobically vs anaerobically.

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Compare aerobic vs anaerobic growth.

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Which tube has an obligate aerobe? An obligate anaerobe?

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What physiological type? obligate aerobe, facultative anaerobe, or obligate anaerobe?

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Acinetobacter Gram stain

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Clostridium sp Gram stain

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Enterococcus sp. from colony on plate

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Candle jar. When flames burns out, ~6% oxygen left.

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Anaerobic jar. Packet removed from foil will be placed in jar to absorb all oxygen.

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Viewed from the bottom, dark red colony indicates lactose fermentation.

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Compare aerobic vs anaerobic growth on nutrient agar

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Compare aerobic vs candle jar growth on nutrient agar.

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Compare candle jar vs anaerobic jar growth on nutrient agar.

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Compare growth on nutrient agar anaerobically, vs subsequent aerobic incubation.

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E. coli appearance, bottom view. Halo of bile salts preciptate around colony.

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No dark centers for colonies, which indicates it cannot ferment lactose.

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No growth from Enterococcus. Gram + bacteria inhibited by crystal violet in MacConkey's agar.

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MacConkey's agar. Clear colonies indicate no fermentation of lactose.

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MacConkey's agar, view from bottom of plate. Dark red colony indicates lactose fermentation, typical of coliform bacteria.

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Coliform bacteria (dark red center) on MacConkey's agar, bottom view.

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MacConkey's agar, with five colonies typical of Escherichia coli.

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Enterococcus growing on azide agar (left), but not on EMB agar (right)

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Gram negative bacterium, growing on EMB agar (right), but not on azide agar (left)

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Compare growth of catalase + and catalase - bacteria on azide agar.

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When examined from the bottom of the Petri dish, Enterobacter will produce a dark-centered colony, indicating lactose fermentation.

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EMB agar, bottom view. Dark centers indicate lactose fermentation, typical of coliform bacteria.

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Bottom view of coliform bacterium. Dark center indicates lactose fermentation.

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Typical green sheen produced by E. coli due to the extreme acid from lactose.

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Measure diameter of zones of inhibition to determine relative sensitivity/resistance to antibiotic.

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Kirby-Bauer Antibiotic Sensitivity Test

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Serratia marcescens

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Effect of 265nm UV light on Serratia marcescens

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Bacteria from fingers

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Staphylococcus epidermidis = white colonies; Micrococcus sp. = yellow colonies

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Environmental contaminants in #1 &2: normal flora in #5 & 6.

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Staphylococcus Gram stain

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Micrococcus Gram stain. Note packets of 4s.

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Streptococcal hemoysis: alpha (top left); beta (right), gamma (bottom)

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Alpha hemolytic streptococci; a green halo around a small colony

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Beta hemolytic streptococci - with complete lysis around a small colony.

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Compare alpha and beta hemolytic streptococci.

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Throat swab on blood agar. Alpha hemolytic streptococci present.

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Streptococcus Gram stain from colony on plate.

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Neisseria sp, Gram - cocci in 2s

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Streptococcus salivarius from a tongue swab, forming capsules and smooth colonies on sucrose-gelatin agar.

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Close up, Streptococcus salivarius colonies with capsule. Non-capsulated Streptococcal colonies are pin-point in size.

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Streptococcus Gram stain from broth.

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Tellurite glycine agar, with Staphyloccus aureus colonies

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Staphylocococcus aureus colonies on Tellurite-glycine agar

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Comparison of Staphylococcus epidermidis (left)  with Staphylococcus aureus (right) on tellurite glycine agar.

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Phage plaques on lawn of host E. coli cells

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Coagulase test with rabbit plasma. Still liquid on left = coagulase negative; clot formation on right = coagulase positive.

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Another view of coagulase positive reaction (bottom) by Staphylococcus aureus

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Coagulase Positive (top) and Negative (bottom)

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Colilert test for both total coliforms and E. coli. If media turns yellow, ONPG +, coliform bacteria present. Beta galactosidase is produced.

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With a long-wave UV light, fluorescence indicates the hydrolysis of MUG by beta glucuronidase of E. coli.

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E. coli count Petrifilms. Coliform bacteria besides E. coli produce red colonies with gas bubbles (from lactose fermentation). E. coli produced blue colonies from glucuronidase action, and gas bubbles from lactose fermentation.

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Can bacteria from Colilert + tube penetrate toilet paper?

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Yes they can! Wash hands after using the restroom.

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EMB colony appearance of three genera of coliform bacteria.

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Klebsiella produces a gummy capsule on EMB.

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Coliform isolate on EMB agar, top appearance.

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Top view of coliform bacterium on EMB agar.

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Sauerkraut fermentation, 7th day.

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Leuconostoc, heterofermenter, first to dominate in sauerkraut

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Sauerkraut,7 days, Gram + rods, Lactobacillus, dominate.

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beaker cheese. Streptococcus lactis starter, rennet added

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curd from beaker cheese

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draining whey from beaker cheese

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Starter culture bacteria, Streptococcus lactis, used to make these cheeses.

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Penicillium camemberti grows on the surface of camembert cheese. The proteases and lipases it produces slowly ripens the cheese. It produces white conidia.

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Penicillium camemberti, the mold which ripens Camembert and Brie cheese from the outside.

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Penicillium roqueforti, which ripens Roquefort and blue cheese.

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Transformation test. BHI agar on left, Minimal lactate agar on right. Juni auxotroph alone in #1, Juni + DNA from wild type Acinetobacter in #2; Juni + DNA from Pseudomonas sp. in #3.

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Conjugation test. Recombinant cells growing on Mineral Glucose agar after conjugation.

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Agglutination reacton. Antigen and antibody combine to form a white precipitation, on the left.

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Lymphocyte has a large nucleus. Responsible for antibody production. Non nucleated red blood cells also present.

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The Monocyte is larger than the lymphocyte, and is phagocytic.

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Segmented nuclei in neutrophils, phagocytic white blood cells.

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