Invertebrate Zoology: Protista

How do single celled organisms make a living?

Thomas Sobat, PIE Fellow, Biology Department, Ball State University, Muncie, Indiana 47306

PRIMARY INDIANA STANDARD ADDRESSED:

5.4.2 Observe and describe that some living things consist of a single cell that needs food, water, air, a way to dispose of waste, and an environment in which to live. Inclusions: 5.4.4, 5.4.5, 5.4.6, 5.5.9, 5.5.10, 5.6.4

OBJECTIVES
Students will understand that there are single celled organisms, and that these organisms are not defined as animals. Students will know that single celled organisms require food, water, air and an environment suitable to maintain life. Students will generate hypotheses regarding the maintenance of protist life in variable environments. Students will have a research experience in which they will test their hypotheses and report their findings

MATERIALS
5 beakers (500-1000 ml) Aquarium pump Aquarium hose 4 air stones 1 four-way gang valve Distilled water Microscope Microscope slides and cover slips Eyedroppers (1/treatment) Petri dishes (1/treatment) Single edged razorblade Club soda or Protoslow Dark box (to cover light beaker)

INTRODUCTION
Background information The protists (Kingdom Protista) are the simplest eukaryotes, yet they represent an incredibly diverse group. Most are unicellular, while others are colonial and closely related to single protist cells. It is the unicellular character that separates protists from the Kingdom Animalia, whose members are multicellular by definition. One group of protists, the algae, are autotrophic (photosynthesizers), while the rest eat bacteria, other protistan cells, or small organic particles suspended or dissolved in water As with all eukaryotic cells, protists contain membrane-bound nuclei and endomembrane systems, as well as numerous organelles. Movement is often provided by one or more flagella, cilia, or by cellular extension (psudopodia), and cilia are often present on the plasma membrane as sensory organelles. Unlike

prokaryotes, protistan nuclei contain multiple DNA strands. Although they are significantly less complex than other eukaryotes, protists can reproduce asexually, and some are capable of meiosis and sexual reproduction. Cellular respiration in the kingdom is primarily an aerobic process (with oxygen), but some protists, including those that live in mud below ponds or in animal digestive tracts, are anaerobes (without oxygen). Protists represent an important step in early evolution, evolving from prokaryotes and eventually giving rise to the entire line of eukaryotes. The first protists probably evolved over 1.7 billion years ago, 2.3 billion years after the origin of life, from simple communities of prokaryotic cells.

The Kingdom Protista is divided into three groups, grouped by lifestyle: the protozoans, the slime molds and algae. Protozoans include all protists that ingest their food and live in aquatic habitats, such as ponds, drops of water in soil, or the digestive tracts of animals.

Slime molds have both unicellular and colonial life stages. When sufficient bacteria (food) are present, cellular slime molds are single cells; however, when food becomes scarce, they aggregate into slug like colonies, which become large reproductive structures.

The third group of protists, the algae, contain chloroplasts and photosynthesize like plants; these can be unicellular or colonial. The algae

inhabit all aquatic environments, and form symbiotic relationships with fungi in terrestrial environments (lichens).

Lesson information The premise by which this lesson has been developed involves the students attempting to determine the requirements of life (oxygen, food, light, etc.) for these seemingly simple organisms. Initially students should be exposed to instructional material that describes the distinguishing characteristics within the protists, and between the protists and the animals. It is imperative that throughout this lesson students are reminded that protists are single celled organisms and not animals. Students will be responsible for developing hypotheses regarding protist life maintenance, by relating their knowledge of plant and animal life to the simpler protist analogues (algae and protozoans). Using simple beaker sized microcosms, students will test their hypotheses with protist farm raised mixed cultures colonized on microscope slides. In the light of good (fair) science students will compare organisms maintained in a control setting to various treatment groups exploring the effect of single variable manipulations. In keeping with the scientific method students will report the results of their experiments in the form of a written manuscript or presentation. This form of assessment will require them to research the topic organisms, manage data, and develop graphics.

PROCEDURE
1.) Following the presentation of pertinent introductory material students should be encourage to develop hypotheses regarding protist life requirements relative to student observations of the requirements of plants and animals.

2.) Requirements (variables) should be tested individually by comparing the effects of the absence of one variable in a treatment microcosm to its presence in the control microcosm following a defined period of time (seven days). Microcosms will be beaker sized, and the control will include protist farm water, an air stone with air supplied through it, and an ambient light source. Variables included (water, food, air, and light) are the fundamental requirements for life in plants, animals and protists, and should be the targets of student hypotheses (variations on the theme can be attempted).

Experimental design Each Microcosm (control and treatments) will have one colonized microscope slide placed inside of it, and allowed to incubate for a given period of time (in excess of six days). Microcosms should be set up as follows (see figure): control - will include protist farm water, an air stone with air supplied through it, and an ambient light source. Food to test the effects of the absence of food, place one colonized slide in a microcosm that contains an air stone and fill it with distilled water.

Light to test the effect of the absence of light, place one colonized slide in a microcosm filled with protist farm water that contains an air stone, and cover it with a light restrictive box. Air - to test the effect of the absence of air, place one colonized slide in a microcosm filled with protist farm water without an air stone. Water to test the effect of the absence of water, place one colonized slide in a microcosm without water

Slide preparation Slides are to be incubated in a protist farm following the design put forth by Wood (1996) for a period in excess of 10 days, and it is imperative that all slides are treated equally. To view colonized slides: 1. Scrape the material from both sides of the slide into a dry Petri dish. 2. Rinse both sides of the slide over the Petri dish using 10 ml of distilled water and mix thoroughly (steps 1 and 2 may be completed up to 24 hours in advance). 3. Place one drop of the protist rich mixture on a clean dry microscope slide, and cover it with a cover slip. 4. Place slide on microscope and view using at least 100x magnification.

3.) Prior to experiment setup one slide should be scraped, viewed and initial counts should be made. Multiple students can complete this, each making and counting their own slides. Individual counts can be used as replicates, and be averaged. Slide counts should proceed as follows: 1. All counts should be completed using the same magnification. 2. Start at one edge of the cover slip and move to the other edge counting all organisms in the field of view (the number of passes may vary based on colonization time. If initial slide counts require x number of passes across the slide to yield >100 organisms, then all counts should be made using x passes.). 3. Students should distinguish between green immobile and non-green mobile cells, and record these counts separately.

4.) After the experiment has been set up and allowed to incubate for a period of at least seven days. Counts should be made for the control and all of the treatments (students can be divided into groups where it is the responsibility of each group to perform counts on a particular treatment). Comparisons can be made between the initial and the control, and the control and individual treatments.

5.) Upon completion of counts, all replicate counts should be averaged, and data should be entered into a Microsoft Excel spreadsheet (this can be done in a number of ways, 1. use calculators, 2. students can calculate averages using their math skills, or 3. all data points can be entered into the spreadsheet and

students can use the CALC AVG function of the software). Students can develop graphs using the graphing function associated with the spreadsheet software (see Excel help). Once graphs have been generated the class should discuss trends in the data so that students have a clear understanding of their results prior to writing their reports.

ASSESSMENT
The experiments can be run to a number of different levels, each step could be considered a unit of assessment. If students are held responsible for hypotheses, experiments and graphs, points could be assigned for each stage. Add report writing or a quiz and the level of assessment increases (see attached quiz).

EXTENSIONS
One variation of this lesson might include adding multiple slides to the control and treatment microcosms upon setup, and continuing the experiment for an extended period. Each week an additional series of slides could be processed, and temporal aspects of this study could be added to the hypotheses.

CONNECTIONS
This lesson would be an outstanding continuation of a microscopy lesson, or a precursor to curriculum dealing with plants and animals.

References Wood, T.S. 1996. Aquarium culture of freshwater invertebrates. The American Biology Teacher. 56(1) 46-50.

Protist Quiz 15 points Name_____________ Match the following organisms with the proper Kingdom 1. 2. 3. 4. 5. ___Algae ___Mouse ___Protozoan ___Oak tree ___Human A. Animalia B. Protista C. Plantae

True False: 6. ___ Protists are multicellular animals 7. ___ Algae are protists 8. ___ Bacteria are protists 9. ___ Some protists make food using the suns energy 10. ___ Some protists eat bacteria 11. The following are all protists except: a. Algae b. Slime mold c. Protozoans d. Worms 12. Protozoans can be found in: a. Ponds b. Soil water c. Digestive tract of an animal d. All of the above 13. Algae a. eat bacteria b. get energy from the sun c. are animals d. are plants 14. ____________ are unicellular, and animals are _______________

GLOSSARY
Aerobic - requiring the presence of air or free oxygen to sustain life. Anaerobic - living in the absence of air or free oxygen. Animalia - the taxonomic kingdom comprising all animals. Autotrophic - any organism capable of self-nourishment by using inorganic materials as a source of nutrients and using photosynthesis or chemosynthesis as a source of energy, as most plants and certain bacteria and protists. Cilia - short, hairlike, rhythmically beating organelles on the surface of certain cells that provide mobility, as in protozoans, or move fluids and particles along ducts in multicellular forms. Eukaryote - any organism with a fundamental cell type containing a distinct membrane-bound nucleus. Flagella - a long lashlike appendage serving as an organ of locomotion in protozoa, sperm cells, etc. Invertebrate - a. without a backbone or spinal column; not vertebrate. b. of or pertaining to creatures without a backbone. Kingdom - a taxonomic category of the highest rank, grouping together all forms of life that share fundamental characteristics Lichen- any complex organism of the group Lichenes, composed of a fungus in symbiotic union with an alga, most commonly forming crusty patches on rocks and trees. Microcosm - a little world; a world in miniature Organelle - a specialized cell structure that has a specific function; a cell organ. Prokaryote - any one-celled organism that lacks a distinct membrane-bound nucleus and has its genetic material in the form of a continuous strand forming loops or coils: characteristic of monerans. Protista - a taxonomic kingdom comprising the protists. Protozoa - any of various one-celled protist organisms that obtain nourishment by ingesting food particles