Kingdom (Eu)bacteria and Simple Eukaryotes (Former K. Protista): Microscopic Identification Goals:

(1) Recognize microscopically and sometimes macroscopically local aquatic microorganisms to Kingdom and Phylum level (2) Identify to family or genus some important local members: e.g., Nostoc, Oscillatoria, Spirogyra, Desmids, Paramecium, Euglena. (3) Understand key characteristics of groups (e.g. prokaryote vs. eukaryote, cell wall characteristics, etc.)

I. KINGDOM (EU)BACTERIA prokaryotic=no nucleus, no membrane-bound organelles

A. Phylum Cyanobacteria cyano =bluegreen (color is very help to ID a cyanobacterium)

1. Examine macroscopic forms of the bacteria Nostoc, Oscillatoria, and Rivularia (if available). Rivularia looks like tiny (1 mm or less) beads of jelly on aquatic plants, especially chara

2. Make wet mounts of Nostoc, Oscillatoria, and Rivularia and examine them using a compound microscope.

3. Nostoc (below)composed of filaments of unicellular bacteria (strings of pearl). Larger football-shaped cells called heterocysts (for fixing N) .

4. Rivularia these look like miniature Nostoc balls (tapioca size) attached to aquatic plants. Under the microscope the filaments radiate outward like spokes..

5. Oscillatoria (right)the filaments are composed of numerous rectangular cells that are barley visible under high power. Sometimes the cells oscillate back and forth like an accordion.

6. Other Cyanobacteria to look for: (a) Spirulinablue green spiral

filaments (b) Komvophoron---like thickened

ms linked together (c) Chlorococcustwo half circles

linked, dark olive 7. Where are the photosynthetic pigments found in

prokaryotes?

B. Miscellaneous heterotrophic bacteriaunder high power, look for the various shapes of bacteria (rods, spirilla, cocci) in water samples from Harlow Lake (next week). Use high power and reduce the light. They will be barely visible.

Historically, many relatively simple, unicellular, eukaryotic (i.e. having a nucleus) organisms were often lumped into an artificial kingdom called Protista. With the many

Kingdom (Eu)bacteria Phylum Cyanobacteria

Nostoc grape- like, gelatinous balls

high magnif. pearl-like chains

Rivularia tiny tapioca like balls on veg

radiating filaments In jelly

Oscillatoria

dark green slime

fine filaments

accordion arrangement

Tiny, narrow rect. cells

Spirulina

Gleocapsa tiny colonies w/mucilage sheath

ALL dark-green (cyan or olive), small (need 400x)

advances in cell and molecular biology in the past decade, many of the protistan groups have been reclassified: some have been absorbed into existing kingdoms, while others have been combined into completely new kingdoms. Many scientists dont bother fitting new groups into the old Linnean hierarchy (Kingdom, Phylum, Class, etc) but rather they simply become clades (literally branches) in an evolutionary tree of life. The following covers these simple eukaryotic organisms formerly classified as protists. II. KINGDOM PLANTAE

A. Phylum Chlorophyta green algae

1. Examine macroscopic form of filamentous green algae. Note how they differ in color from macroscopic Oscillatoria? 2. Prepare a wet mount of the filamentous green algae. Note the size of the cells (relative to Cynanobacteria) and the presence of chloroplasts. Some filamentous green algae have conspicuously shaped chloroplasts, such as Spirogyra (spiral shaped) and Zygnema (star-shaped). Cladophora is a common branched filamentous form on rocks in local streams. 3. Desmids are green algae composed of 2 semi-cells (mirror image halves). Cosmarium (below, left) resembles tiny, bright green butterflies. Micrasterias (below, mid) resembles a snowflake. Closterium (right) which is common in Harlow Lake, is shaped like a bright green banana.

4. Tiny, green Carteria (right) or Chlamydomonas (similar but with 2 flagella) may be seen moving rapidly across the slide. These are flagellated green algae, but the flagella are hard to see in live preparations. 5. Often Chlamy-like cells are organized into spherical colonies, such as Volvox (1000s of cells), or Gonium (few cells)

6. Chara (stonewort) looks like an aquatic plant, but is actually a green alga with huge cells (some species have cells >2 inches long!).

Kingdom Plantae) Phylum Chlorophyta (green algae)

yellow green slime- filaments visible to naked eye

cells lined up in long filaments

cell detail easily visible under low power

Spirogyra

Zygnema

Pediastrum Chlamydomonas

tiny, green flagellated highly motile! Chara

(stonewort) looks like pond- weed

super cells 1-2 long

Desmids: the 3 above are green algae w/ mirror image semi-cells. They are all bright green unlike diatoms, which are golden brown inside

Closterium crescent moon

Micrasterias

Cosmarium

Filamentous green algae (various species)

Phylum Chlorophyta (continued)

Volvox aureus

Gonium formosum

Eudorina

Gonium sociale

Gonium pectorale

Pleodorina illinoisensis

Mastigosphaera

Pandorina morum

Stephanospaera

III. KINGDOM (or Clade) STRAMENOPILA (strameno=straw pila=hair) Stramenopila is new clade composed of very different looking organisms: diatoms, brown algae, and water molds/blights. In some reproductive stages, they all possess a biflagellated spore with one flagellum being hairy. Diatoms, which are among the smallest eukaryotes seem to have little in common with kelp seaweeds (among the largest eukaryotes), but they both possess the same unique photosynthetic pigments and also share a similar, unique storage carbohydrate.

A. PHYLUM CHRYSOPHYTA (DIATOMS) chrys=gold

1. Look for diatoms such as Amphora (fig. 516), Asterionella (fig. 504), Cyclotella (fig. 489), Rhopalodia (fig. 526), Navicula (fig. 542), & Tabellaria (fig. 508). Be sure to be able to distinguish diatoms from Desmids (see Green Algae) on the basis of chloroplast color. Diatom chloroplasts are golden brown. Diatom cell walls are composed of silica (literally glass). Their clear, often sculptured shells make diatoms easy to spot. Some diatoms move slowly by jet propulsion (expelling water through holes in their shell) and resemble tiny ships.

arms usually fall off and are seen separate

IV. KINGDOM (or Clade) ALVEOLATA The new clade Alveolata is composed of dinoflagellates, ciliates, and apicomplexa

(malaria, etc.). All have little sacks (alveoli) just underneath their cell membranes. Surprisingly, the function of alveoli in these organisms is poorly known.

A. PHYLUM DINOFLAGELLATA (dinoflagellates) dyno refers to

spinning/moving in a circle. Under high power, looks for small reddish brown spherical algae, possibly moving with a spinning action.

Kingdom Alveolata Phylum Pyrophyta (Dinoflagellates)

All very tiny, reddish brown; SPINNING motion is good for ID!

8-9 Ceratium

4-6=Gymnodium spp. 1-3 =Glenodinium spp.

7-8 Peridinium

B. KINGDOM ALVEOLATA: PHYLUM CILIOPHORA=bearing cilia

1. Prepare wet mounts of stock solutions or soup samples collected from Harlow Lake and search for ciliates, named for rows of cilia covering their bodies. The most common forms will include Parmecium, Coleps, Euplodes, and stalked forms, such as Vorticella and Epistylis, which attach to weed or sand.

Kingdom Alveolata Phylum Ciliophora (Ciliates)

Euplotes

Vorticella

colonial ciliate-Epistylis

Coleps

Paramecium spp.

Stentor

V. Kingdom (or Clade) Euglenozoa This new clade includes most of the protozoans that possess one to many long flagella, and usually have a flexible proteins covering called a pellicle. Within this kingdom there are two main phyla: Phy. Euglenophyta, many of which are green and possess chloroplasts (see examples below), and Phy. Kinetoplastida, a diverse group of heterotrophic (non-photosynthetic) flagellates, some of which are important parasites.

VI Kingdom or Clade Amoebozoa .

Many amoeboid (amoeba-like) organisms were lumped together in the past, but molecular evidence indicates that the amoeboid like morphology probably arose more than once and some amoebas once lumped to together, really belong in separate clades that may only be distantly related. Rhizopods (common garden variety of Amoeba) and slime molds (once considered fungi) belong to the clade Amoebozoa.

VII Kingdom or Clade Rhizaria

These are amoeboid organisms that have very fine threads of protoplasm (pseudopodia) that extend into the environment through pores in their shells, which are made of silica or calcium carbonate. Actinopods (lower right) look like rays of light radiating from a glass sphere, and are sometimes called Heliozoans or sun animals

Ameoba sp. (Rhizopod) Difflugia (rhizopod w/shell) Actinosphaerium (Rhizaria)

Kingdom (Eu)bacteria and Simple Eukaryotes (Former K. Protista): Microscopic IdentificationGoals:(1) Recognize microscopically and sometimes macroscopically local aquatic microorganisms to Kingdom and Phylum level(2) Identify to family or genus some important local members: e.g., Nostoc, Oscillatoria, Spirogyra, Desmids, Paramecium, Euglena.(3) Understand key characteristics of groups (e.g. prokaryote vs. eukaryote, cell wall characteristics, etc.)