LAB 2 HOMEWORK AMBER M - FINAL

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Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ LAB 2 HOMEWORK – AMBER MCCLENDON Exercise 3-3: Examination of Eukaryotic Microbes Terms: Plastid: A double membrane bound organelle involved in the synthesis and storage of food that is commonly found within the cells of photosynthetic organisms like plants. Its function largely depends on the presence of pigments. A plastid involved in food synthesis typically contains pigments, which are also the ones responsible for the color of a plant structure (e.g. green leaf, red flower, yellow fruit, etc). A plastid containing green pigment (chlorophyll) is called a chloroplast whereas a plastid containing pigments apart from green is called a chromoplast. A plastid that lacks pigments is called a leucoplast, and it mainly is involved in food storage. A leucoplast may be an amyloplast that stores starch, an elaioplast that stores fat, or a proteinoplast that stores proteins. Like mitochondria, plastids have their own DNA and ribosomes; hence, they may be used in phylogenetic studies. Source: http://www.biology-online.org/dictionary/Plastid 8/9/13 Prokaryotes: A small, single-celled organism that lacks a nucleus (instead, it has circular, covalently-closed DNA within the cytoplasm) and other organelles. Its ribosomal components are similar, but somewhat smaller, than those of eukaryotes. Prokaryotes are more primitive than eukaryotes. Eukaryotes: A larger type of cell that contains organelles including a nucleus for its genetic material (DNA). Its DNA is organized in linear strands and often contains introns (intervening sequences). Ribosomal components are similar, but somewhat larger, than their prokaryotic counterparts. Eukaryotes evolved after prokaryotes. Domains: A broad classification of ALL organisms based on the sequence of nucleotides in rRNA. There are three domains: Bacteria and Archaea are two domains within the prokaryotes and Eukarya is the third domain which includes all eukaryotes. rRNA: Ribonucleic acid, the intermediary between genetic information (DNA) and enzymes (protein). That is, gene expression requires the copying of selective portions of DNA into RNA (which is called messenger RNA or mRNA). RNA can also be structural as in ribosomal RNA (rRNA), and it is vitally important in the translation of mRNA into proteins as transfer RNA (tRNA). Supergroups: Five divisions [six divisions in some schemes] of Eukarya that are larger (more inclusive) than the classical Kingdoms. Endosymbiotic theory: The theory whereby a primitive prokaryote was engulfed (but not digested) by a larger prokaryote that led to the formation of eukaryotic cells. This theory is especially well accepted for the case of Archaeplastida where the modern-day chloroplasts of plants and red/green algae are most likely derived from an engulfed cyanobacterium. Trophozoite: Feeding stage in the life cycle of certain protozoans. Cyst: Resting (or dormant) stage in the life cycle of certain protozoans. Autotroph: An organism that fixes CO2 (generates large, complex organic molecules from CO2). Heterotroph: An organism that requires carbon in the form of organic molecules.

Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ Mixotroph: An organism that can live autotrophically (when light is available) or heterotrophically. Historically, Euglena is an important example. Pseudopods: “False feet” or extensions of cytoplasm from amoebae that are used for movement as well as engulfing prey. Amoebae are irreg ularly-shaped organisms that constantly change shape via the production of pseudopods. Cilia: Small filamentous organelles, like fibers, that cover the surface of some alveolates (within the Supergroup Chromalveolata). Cilia are usually numerous whip-like flagella that are used for both locomotion and feeding (directing food particles through the oral groove to the cytosome). Saprophyte(s): A heterotroph that digests dead organic matter (a decomposer). Parasite(s): An organism that lives symbiotically with another, but to the detriment of the other organism (the host). Mold(s): An informal grouping of filamentous fungi. Yeast: An informal grouping of unicellular fungi. Hypha(e): A filament (or filaments) of fungal cells. Mycelium: A mass of fungal filaments (hyphae). Conidia: The asexual, non-motile spores of a fungus. They are the small spheres that appear at the ends of the fruiting bodies of both Aspergillus and Penicillium molds. Conidia are dispersed when they become airborne something like the seed heads of a mature dandelion. Zygospore(s): Product of fertilization and the site of meiosis in some molds. Biol 212 Exercise 3-3: Examination of Eukaryotic Microbes Fill in the blanks and answer questions:

Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ I. Theory A. Prokaryotes vs. Eukaryotes (Table 3-3) 1. Prokaryotes: two domains- Archaea & Bacteria 2. Eukaryotes: one domain - Eukarya a) Kingdoms: Plants, Animals, Protists & Fungi (1) Microscopic – Protists & Fungi (a) Protists (protozoans & algae) (b) Fungi (yeasts & molds) II. Supergroup Excavata A. Unicellular, feeding groove, one or more flagella. B. Typical lifecycle typically includes both trophozoite & cyst. 1. Subgroup: Parabasalids a) Representative: 1. Trichomonas vaginalis Nucleus, Flagella, Undulating membrane, Hydrogenosomes Which sexually transmitted disease (STD) is cause by this organism? Trichomoniasis, Which can cause inflammation of the vulva, or vagina known as vulvovaginitis 2. Subgroup: Diplomonads a) Representative: 1. Giardia lamblia - trophozoite Mitosomes , Paired nuclei, Flagella (four pairs), Median bodies (2) In figure 3-13 Panel B can you see the four pairs of flagella? No 3. Subgroup: Euglenozoans a) Representative: 1. Euglena sp. Green, Photosynthetic, Mixotrophic, Flagella (1/2), Eyespot What structure gives these organisms their photosynthetic ability? Chloroplasts 4. Subgroup: Kinetoplastids a) Representative: 1. Trypanosoma brucei (gambiense or rhodesiense) (do not have slides in lab) One large mitochondrion What disease does this parasite cause? 2. Trypanosoma cruzi (slides we have in lab) Causes Chagas Disease, Throughout south and central America

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Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ T. brucei gambiense is the main cause (≥ 98% of cases) of African sleeping sickness in sub -Saharan Africa. It is a neurological disease that commonly leads to meningitis (inflammation of the protective membranes covering the brain and spinal cord) (We didn’t look at this spe cies in the lab) This disease is transmitted by the Testi fly. T. cruzi targets the heart and gastrointestinal tract and causes Chagas Disease throughout South and Central America III. Supergroup: Archaeplastida A. Chloroplasts, Autotrophic, Cellulose cell walls 1. Subgroup: Chlorophyta (Green algae) Freshwater, unicellular, filamentous, or colonial, flagella (common) a) Representatives: (1) Chlamydomonas sp. Haploid, unicellular, chloroplast, flagella, stigma (2) Volvox sp. (prepared slide) Colonial, sexual reproduction, male (sperm bundles) and female colonies (eggs) Identify colony, daughter colonies Daughter colonies are the result of what type of reproduction? Asexual 2. Subgroup: Charophytes Close relatives to plants a) Representative: (1) Spirogyra sp. (prepared slide) Filamentous, haploid, conjugation, zygospore (sexual reproduction, diploid) Identify chloroplasts (spiral), nucleus, cell wall IV. Supergroup: Chromalveolata A. Got their plastids from an algal cell ancestor 1. Subgroup: Alveolates Heterotrophic or autotrophic, small membrane sacs beneath their cytoplasmic membrane a) Ciliates - Cilia, oral groove, cytosome Representatives: (1) Paramecium sp. (prepared slide & live culture if available) Identify macronucleus, cilia, oral groove and contractile vacuole (if live culture is available) (2) Stentor sp. Cilia, beaded macronucleus, cytosome, green (3) Balantidium coli - trophozoite (prepared slides) Identify cilia and macronucleus

Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ What are the symptoms of an acute infection cause by this organism? Acute infection: bloody, mucoid stool (diarrhea as often as every 20 minutes) Chronic infection: alternating diarrhea and constipation b) Apicomplexa Non-motile Complicated life cycles, with more than one host, affecting multiple tissues Representative: (1) Plasmodium falciparum (prepared slide) In humans it infects liver and RBCs; in mosquitoes it infects gut and salivary glands Identify human red blood cells and parasites. Find a red blood cell with the parasite on the inside of the cell. What disease does this organism cause in humans? Malaria

Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ 2. Subgroup: Stramenophiles (also spelled Stramenopiles) Most have a unique flagellum with three-parted lateral hairs a) Diatoms Lack flagella, aquatic, single celled or colonial autotrophs Fucoxanthin is a golden brown pigment, cell walls have two halves with one half overlapping the other Representative: (1) Diatoms – mixed species (prepared slide) Identify centric and pennate forms What inorganic compound gives a glass-like property to their cell walls? Silica (silicon dioxide, SiO2) V. Supergroup: Unikonta A. Heterotrophs (amoebas, animals, fungi, and others) 1. Subgroup: Gymnamoebas Representative: (1) Amoeba proteus (prepared slide & live culture if available) Identify nucleus & pseudopods 2. Subgroup: Entamoebas Representative: (1) Entamoeba histolytica - trophozoite (prepared slides) Identify nucleus Describe what the nucleus looks like: Ring around the nucleus (looks like a “bullseye” of a target) What type of specimen is used to diagnose infections caused by this organism? Stool sample 3. Subgroup: Fungi a) Yeasts Representatives: (1) Saccharomyces cerevisiae (prepared slide) Identify vegetative cells, nuclei, and budding cells Does this yeast cause diseases in humans? NO, S. cerevisiae is often called baker’s or brewer’s yeast because it is used in the making of bread, beer and wine. (2) Candida albicans (prepared slide) Identify vegetative cells and budding cells Define pseudohyphae : A chain of easily disruptedfungal cells that is intermediate between a chain of budding cells and a true hypha. Is this organism an opportunistic pathogen? Yes, C. albicans can cause thrush in the oral cavity, vulvovaginitis or cutaneous infection of skin. b) Molds Representatives: (1) Rhizopus sp. (two prepared slides) Identify asexual spores, sexual spore (zygospore) On what type of food is this genus of mold commonly found? Bread (2) Aspergillus sp. (prepared slide) Identify hyphae, conidia (asexual spores)

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Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ What commercial products are manufactured using this organism? Soy paste and soy sauce from soybeans (A. oryzae & A. soyae) & Citric acid (Aspergillus niger) (3) Penicillium sp. (prepared slide) Identify hyphae, conidia (asexual spores) What food product is manufactured using this organism? Blue-veined cheeses (ex P. roquefortii and P. camemberti in Roquefort, Camembert and Brie) Sausage and ham (ex P. nalgiovense to improve flavor and to prevent colonization by other molds and bacteria) ******On the next three pages draw and label pictures of each using your lab manual and/or PPT****** Domain Eukarya Domain Eukarya Lab slides are poor refer to picture in your book Supergroup Excavata Supergroup Excavata Subgroup Parabasalids Subgroup Diplomonads Name Trichomonas vaginalis Name Giardia lamblia Identify nucleus & flagella Identify Trophozoite- paired nuclei Total Magnification 1,000x (oil) Total Magnification 1,000x (oil) Trophozoite: Vegetative “feeding” stage of life cycle Domain Eukarya Domain Eukarya Supergroup Excavata Supergroup Excavata Subgroup Euglenozoans Subgroup Kinetoplastids Name Euglena Name Trypanosoma Cruzi Identify Identify nucleus & kinetoplast Total Total 1,000x (oil)

Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ Magnification Lab slides are poor refer to picture in your book Magnification Domain Eukarya Domain Eukarya Small individuals make up the spheres Supergroup Archaeplastida Supergroup Archaeplastida Subgroup Chlorophyta (Green algae) Subgroup Chlorophyta (Green algae) Name Chlamydomonas Name Volvox Identify Flagella Identify daughter colonies Total Magnification 1,000x (oil) Total Magnification 100x (low) Domain Eukarya Domain Eukarya Supergroup Archaeplastida Supergroup Archaeplastida Subgroup Charophytes Subgroup Charophytes Name Spirogyra † Name Spirogyra † Conjugation Identify cell wall, chloroplasts, nucleus Identify zygospore Total Magnification 100x (low) Total Magnification 100x (low) † Note there are two slides for spirogyra

Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ Domain Eukarya Domain Eukarya Note the “kidney - shaped” nucleus best seen at 400x Supergroup Chromalveolata Supergroup Chromalveolata Subgroup Alveolates Subgroup Alveolates Name Paramecium Name Balantidium coli Identify macronucleus & cilia* Identify Trophozoite - nucleus & cilia* Total Magnification 100x (low) Total Magnification 100x (low) *If you fine focus up and down you can sometimes see the cilia around the edge of the cell Domain Eukarya Lab slides are poor refer to picture in your book Domain Eukarya Notice the organisms are found inside and outside the red blood cells Supergroup Chromalveolata Supergroup Chromalveolata Subgroup Alveolates Subgroup Alveolates Name Stentor Name Plasmodium falciparum Identify beaded macronucleus Identify Red blood cells & parasites Total Magnification 100x (low) Total Magnification 1,000x (oil) Domain Eukarya best seen at 400x Supergroup Chromalveolata Subgroup Stramenophiles Name Diatoms Identify centric and pennate forms Total Magnification 100x (low) Domain Eukarya Domain Eukarya Supergroup Unikonta Supergroup Unikonta Subgroup Gymnamoebas Subgroup Entamoebas Name Amoeba Name Entamoeba histolytica

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Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ Identify nucleus & pseudopods Identify Trophozoite nucleus Notice the unique “bullseye” nucleus Lab slides are poor refer to picture in your book Total Magnification 100x (low) Total Magnification 1,000x (oil) Pseudopods: cytoplasmic extensions used to move Domain Eukarya Domain Eukarya Pseudohyphae*: look like hyphae but are not true hyphae. They form when budding cells fail to separate. Supergroup Unikonta Supergroup Unikonta Subgroup Fungi (unicellular yeast) Subgroup Fungi (unicellular yeast) Name Saccharomyce s cerevisiae Name Candida albicans Identify vegetative cells, budding cells Identify Vegetative, budding cells, & pseudohyphae Total Magnification 1,000x (oil) Total Magnification 1,000x (oil) *seen only on the old slides (round cover slip) Domain Eukarya Domain Eukarya Supergroup Unikonta Supergroup Unikonta Subgroup Fungi (filamentous molds) Subgroup Fungi (filamentous molds)

Lab 2 (Examination of Eukaryotic Microbes) N ame:_____________________________ Name Aspergillus (wm) Not pictured in your book as whole sample Name Aspergillus (sec) Identify conidia (asexual spores) Hyphae Identify Conidia (asexual spores) Total Magnification 100x (low) Total Magnification 100x (low) whole (wm) cross section slide (Sec) As seen in your book Domain Eukarya Domain Eukarya Supergroup Unikonta Supergroup Unikonta Subgroup Fungi (filamentous molds) Subgroup Fungi (filamentous molds) Name Rhizopus Name Rhizopus Identify asexual sporangia (sporangiospore s, columella) Identify sexual conjugation (zygospore) Total Magnification 40x (scanning) Total Magnification 100x (low) Domain Eukarya Domain Eukarya Supergroup Unikonta Supergroup Unikonta Subgroup Fungi (filamentous molds) Subgroup (filamentous molds) Name Penicillium Name Unknown‡ Identify Hyphae*, conidia (asexual spores) Identify hyphae Total Magnification 100x (low) Total Magnification ‡ This specimen is from an agar plate; look at the plate then look at the sample under the microscope. (Notice how the hyphae extend throughout the agar. Above the agar you see aerial hyphae upon which asexual reproductive structures will form.)

LAB 2 HOMEWORK AMBER M - FINAL

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