1. UNIT 2 SEMINAR: Observation of Microorganisms
Fundamentals of Microbiology
Microbiology
4/25/2017
UNIT 2 SEMINAR: Observation of Microorganisms
Hi everyone! Welcome to our first seminar! It is great to have you all here today. We will be starting up in a few minutes.
While you are waiting to get started this evening, please take this opportunity to chat and adjust your speaker volumes. Think about the topic for today and the questions assigned in the seminar page.
If you encounter any technical issues accessing a KHE Seminar, please call KU ACE Help , option 2, then option 1. You may also KU ACE Help at
Evelyn I. Milian
Instructor
2011
Evelyn I. Milian - Instructor

2. Evelyn I. Milian - Instructor
Microbiology
4/25/2017
Agenda
Discussion of unit 2 seminar topic:
Observation of microorganisms
Question and answer session (~last 5 minutes):
Course syllabus, assignments, grading, requirements.
* Students in sections with other instructors are also invited to stay for this session; however, specific questions about assignments or grading should be addressed to your instructor.
Hi everyone! Welcome to our Unit 2 seminar! I’m glad you were able to join us! We will start up in a few minutes.
While you are waiting to get started this evening, please take this opportunity to chat and adjust your speaker volumes. In addition, think about the topic for today and the questions assigned in the seminar page.
If you encounter any technical issues accessing a KHE Seminar, please call KU ACE Help , option 2, then option 1. You may also KU ACE Help at
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

3. Questions Assigned for Discussion (Unit 2 Seminar Page)
Microbiology
4/25/2017
Questions Assigned for Discussion (Unit 2 Seminar Page)
Give 3 examples of safety procedures you should always follow in the microbiology lab.
Why do you stain cells before observing them under the microscope?
Name some common shapes and arrangements observed when viewing bacteria under a microscope.
In a Gram stain, one step could be omitted and still allow differentiation between gram positive and gram negative cells. What is that one step?
Assume that you are viewing a Gram-stained sample of vaginal discharge. Large (10um) nucleated red cells are coated with small (0.5 um X 1.5 um) blue- purple cells on their surfaces. What is the most likely explanation for the red cells and blue cells?
A sputum sample from Calle, a 30-year-old Asian elephant, was smeared onto a slide and air dried. The smear was fixed, covered with carbolfuchsin, and heated for 5 minutes. After washing with water, acid alcohol was placed on the smear for 30 seconds. Finally, the smear was stained with methylene blue for 30 seconds, washed with water, and dried. On examination at 1000X, the zoo veterinarian saw red rods on the slide. What infection do the results suggest? (Calle was treated and recovered.)
Discuss 2 other examples of staining techniques, and when they are used.
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

4. Basic Safety Procedures in the Microbiology Lab
The microbiology laboratory is unique in many ways compared to other biological laboratories. While safety is paramount in any lab, the added complication of unseen contaminants and potential hazards must be addressed.
Microorganisms are normally present in the environment, and many kinds are also found on and in the human body.
Improper handling of chemicals, equipment and/or microbial cultures is dangerous and can result in injury or infection.
Avoiding contamination and possible infection warrants stringent safety practices at all times.
Give 3 examples of safety procedures you should always follow in the microbiology lab.
2011
Evelyn I. Milian - Instructor

5. Safety in the Microbiology Lab is Priority!
2011
Evelyn I. Milian - Instructor

6. Basic Safety Procedures in the Microbiology Lab
Do not eat, drink, smoke, or bring food or drinks into the lab room.
Do not apply cosmetics or handle contact lenses in the lab.
Wear protective clothing (i.e., a lab coat) and closed-toed shoes in lab.
Tie back long hair, as it is a potential source of contamination as well as a likely target for fire.
Do not work with an uncovered open cut or wound. Protect it with a bandage and wear plastic gloves.
Wipe your work area or bench with disinfectant before and after work.
Keep all sources of possible contamination out of your mouth and hands: pencils, laboratory ware and utensils, and other items.
Discard contaminated items—pipettes, Petri dishes, test tubes, and other items—in the designated containers (many times labeled “biohazard”).
Practice aseptic technique at all times when dealing with microbial cultures.
Wash your hands thoroughly for 20 seconds with soap and water after handling living microbes and before leaving the laboratory.
2011
Evelyn I. Milian - Instructor

7. Laboratory Safety Items and Equipment
2010
Biology I - Lab Test 1 - Prof. Evelyn I. Milian

8. Staining Techniques to Observe Microorganisms
Observing microorganisms also presents a special challenge to microbiologists, and to answer that challenge there are a vast array of staining techniques available to enhance observations. There are special staining techniques that take the physiology of the target cells into account.
Why do you stain cells before observing them under the microscope?
Name some common shapes and arrangements observed when viewing bacteria under a microscope.
In a Gram stain, one step could be omitted and still allow differentiation between gram positive and gram negative cells. What is that one step?
2011
Evelyn I. Milian - Instructor

9. Staining Techniques to Observe Microorganisms
Medical Microbiology
2010
Staining Techniques to Observe Microorganisms
Staining increases contrast.
Contrast is the effect of a striking difference, as in color or tone, of adjacent parts (for example, in a photograph or image). It is based on the differential absorption of light by parts of the specimen.
Microscopists improve contrast by coloring specimens with stains (dyes) that bind to cellular structures and absorb light to provide contrast.
Live or unstained cells have little contrast with the surrounding medium. However, researchers do make discoveries about cell behavior looking at live specimens.
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

10. Bacterial Shapes and Arrangements
Microbiology
4/25/2017
Bacterial Shapes and Arrangements
Most common shapes
Bacillus (rod-shaped; bacilli in plural)
Coccus (spherical; cocci in plural)
Spiral (corkscrew or curved)
There are variations to these basic shapes.
Coccobacillus (oval-shaped)
Other less common shapes:
Square
Star-shaped
Cell arrangements: The cells may form groups.
Pairs
Chains
Clusters
Average size: µm wide  2-8 µm long
Bacillus: rodlike
Coccus: spherical or ovoid
Spiral: corkscrew or curved
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

11. Prokaryotic Cells: Bacterial Shapes
BIOLOGY II
4/25/2017
Prokaryotic Cells: Bacterial Shapes
Bacillus (plural: bacilli)
Rod-shaped cells.
Arrangements: pairs, chains.
FIGURE 21.11
a. Spirillum, a spiral-shaped bacterium. b. Bacilli, rod-shaped bacteria. c. Cocci, round bacteria.
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

12. Prokaryotic Cells: Bacterial Shapes
BIOLOGY II
4/25/2017
Prokaryotic Cells: Bacterial Shapes
Coccus (plural: cocci)
Spherical cells
Groups arrangements:
Diplococci (pairs)
Streptococci (chains)
Tetrad (4 cells in cube)
Sarcinae (8 cells in cube)
Staphylococci (grapelike clusters of many cells)
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

13. Evelyn I. Milian - Instructor
Microbiology
4/25/2017
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

14. The Gram Stain: A Differential Stain
Microbiology
4/25/2017
The Gram Stain: A Differential Stain
In a Gram stain, one step could be omitted and still allow differentiation between gram positive and gram negative cells. What is that one step?
THE GRAM STAIN (Tortora, 10 Ed., Chapter 3)
Differential Stains
Unlike simple stains, differential stains react differently with different kinds of bacteria and thus can be used to distinguish them. The differential stains most frequently used for bacteria are the Gram stain and the acid-fast stain.
Gram Stain
The Gram stain was developed in 1884 by the Danish bacteriologist Hans Christian Gram. It is one of the most useful staining procedures because it classifies bacteria into two large groups: gram-positive and gram-negative.
In this procedure (Figure 3.12a),
A heat-fixed smear is covered with a basic purple dye, usually crystal violet. Because the purple stain imparts its color to all cells, it is referred to as a primary stain.
After a short time, the purple dye is washed off, and the smear is covered with iodine, a mordant.When the iodine is washed off, both gram-positive and gram-negative bacteria appear dark violet or purple.
Next, the slide is washed with alcohol or an alcohol-acetone solution. This solution is a decolorizing agent, which removes the purple from the cells of some species but not from others.
The alcohol is rinsed off, and the slide is then stained with safranin, a basic red dye. The smear is washed again, blotted dry, and examined microscopically.
Counterstain: Stains that have a contrasting color to the primary stain; for example: safranin.
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

15. Keiser University - Fort Lauderdale, Florida
4/25/2017
GRAM STAIN (Tortora, Chapter 3)
The purple dye and the iodine combine in the cytoplasm of each bacterium and color it dark violet or purple. Bacteria that retain this color after the alcohol has attempted to decolorize them are classified as gram-positive; bacteria that lose the dark violet or purple color after decolorization are classified as gram-negative (Figure 3.12b). Because gram-negative bacteria are colorless after the alcohol wash, they are no longer visible. This is why the basic dye safranin is applied; it turns the gram-negative bacteria pink. Stains such as safranin that have a contrasting color to the primary stain are called counterstains. Because gram-positive bacteria retain the original purple stain, they are not affected by the safranin counterstain.
As you will see in Chapter 4, different kinds of bacteria react differently to the Gram stain because structural differences in their cell walls affect the retention or escape of a combination of crystal violet and iodine, called the crystal violet–iodine (CV–I) complex. Among other differences, gram-positive bacteria have a
thicker peptidoglycan (disaccharides and amino acids) cell wall than gram-negative bacteria. In addition, gram-negative bacteria contain a layer of lipopolysaccharide (lipids and polysaccharides) as part of their cell wall (see Figure 4.13, page 86). When applied to both gram-positive and gram-negative cells, crystal violet and then iodine readily enter the cells. Inside the cells, the crystal violet and iodine combine to form CV–I. This complex is larger than the crystal violet molecule that entered the cells, and, because of its size, it cannot be washed out of the intact peptidoglycan layer of gram-positive cells by alcohol. Consequently, gram-positive cells retain the color of the crystal violet dye. In gram-negative cells, however, the alcohol wash disrupts the outer lipopolysaccharide layer, and the CV–I complex is washed out through the thin layer of peptidoglycan. As a result, gram-negative cells are colorless until counterstained with safranin, after which they are pink.
2011
Evelyn I. Milian - Instructor
MCB-2000C: Microbiology - Prof. Evelyn I. Milian

16. Differential Stains: Gram Stain
Microbiology
2011
Differential Stains: Gram Stain
In a Gram stain, one step could be omitted and still allow differentiation between gram positive and gram negative cells. What is that one step?
Answer:
The counterstain (or secondary stain) = safranin.
GRAM STAIN (Tortora, Chapter 3)
In summary, gram-positive cells retain the dye and remain purple. Gram-negative cells do not retain the dye; they are colorless until counterstained with a red dye. The Gram method is one of the most important staining techniques in medical microbiology. But Gram staining results are not universally applicable, because some bacterial cells stain poorly or not at all. The Gram reaction is most consistent when it is used on young, growing bacteria.
The Gram reaction of a bacterium can provide valuable information for the treatment of disease.Gram-positive bacteria tend to be killed easily by penicillins and cephalosporins. Gram-negative bacteria are generally more resistant because the antibiotics cannot penetrate the lipopolysaccharide layer. Some resistance to these antibiotics among both gram-positive and gram-negative bacteria is due to bacterial inactivation of the antibiotics.
2011
Evelyn I. Milian - Instructor
Prof. Evelyn I. Milian

17. The Gram Stain and the Bacterial Cell Wall
Microbiology
4/25/2017
The Gram Stain and the Bacterial Cell Wall
In the Gram staining technique, samples are first stained with crystal violet dye and iodine (a mordant that helps retain the stain), then rinsed with alcohol, and finally counterstained with a red dye such as safranin. * The structure of a bacterium’s cell wall determines the staining response. *
CELL WALL
It surrounds the underlying plasma (cytoplasmic) membrane (the plasma membrane is involved in passage of materials into and out of the cell, among other functions).
STEP
Color of
Gram + cells
Gram – cells
Primary stain:
Crystal violet
Purple
Mordant:
Iodine
Decolorizing agent:
Alcohol-acetone
Colorless
Counterstain:
Safranin
Red
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

18. GRAM-POSITIVE BACTERIA GRAM-NEGATIVE BACTERIA
Microbiology
4/25/2017
The Prokaryotic Cell Envelope: Bacterial Classification According to Their Cell Wall Structure and Gram Stain Properties
GRAM-POSITIVE BACTERIA
GRAM-NEGATIVE BACTERIA
Envelope with two layers: cell wall and cell (cytoplasmic) membrane.
Envelope with three layers: outer membrane, cell wall and cell membrane.
Retain the Gram stain (crystal violet) due to their cell wall structure; purple-colored (which is applied first in the procedure).
Do not retain the Gram stain (crystal violet) and will appear pink-red after the counterstain (safranin) is applied.
Cell wall with thick (multilayered) peptidoglycan, containing teichoic acids.
Cell wall with thin (single-layered) peptidoglycan layer. No teichoic acids.
No outer membrane; no LPS, low lipid and lipoprotein content.
Protective outer membrane with LPS (lipopolysaccharides), lipoproteins, and phospholipids.
Their cell wall is almost completely destroyed by lysozyme (a digestive enzyme in eukaryotic cells, found in mucus, saliva and tears).
Their cell wall usually is not destroyed by lysozyme to the same extent as in gram-positive cells; some of the outer membrane also remains.
Highly susceptible to penicillin and sulfonamide (antimicrobial agents).
Low susceptibility to penicillin and sulfonamide.
Peptidoglycan = murein: Un polímero que contiene cadenas alternantes de dos polisacáridos: N-acetylglucosamine (NAG) y N-acetylmuramic acid (NAM) y varios amino ácidos.
GRAM STAIN MECHANISM
Crystal violet-iodine crystals form in cell
Gram-positive
Alcohol dehydrates peptidoglycan
Crystal-violet stain/iodine (CV-I) complex crystals do not leave
Gram-negative
Alcohol dissolves outer membrane and leaves holes in peptidoglycan
CV-I washes out
GRAM POSITIVE CELL WALL
Teichoic acids:
Lipoteichoic acid links to plasma membrane
Wall teichoic acid links to peptidoglycan
May regulate movement of cations
Polysaccharides provide antigenic variation
GRAM NEGATIVE CELL WALL
Lipopolysaccharides, lipoproteins, phospholipids.
Forms the periplasm between the outer membrane and the plasma membrane.
Protection from phagocytes, complement, antibiotics.
O polysaccharide antigen, e.g., E. coli O157:H7.
Lipid A is an endotoxin.
Porins (proteins) form channels through membrane
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

19. Staining Techniques: Clinical Applications (Chapter 3)
Assume that you are viewing a Gram-stained sample of vaginal discharge. Large (10um) nucleated red cells are coated with small (0.5 um x 1.5 um) blue-purple cells on their surfaces.
What is the most likely explanation for the red cells and blue cells?
2011
Evelyn I. Milian - Instructor

20. Staining Techniques: Clinical Applications (Chapter 3)
Microbiology
4/25/2017
Staining Techniques: Clinical Applications (Chapter 3)
The large red cell is an epithelial cell from the vaginal tissue.
Blue-purple cells are gram- positive bacteria; there are also some gram-variable bacilli (Gardnerella vaginalis).
This is bacterial vaginosis (BV), an infection caused by Gardnerella vaginalis.
Vaginosis is most likely a result of a shift from a predominance of “good bacteria” (lactobacilli) in the vagina to a predominance of “bad bacteria”.
BACTERIAL VAGINOSIS – Gardnerella species (Ref. Cowan’s book, Ch. 23)
Infection called vaginosis rather than vaginitis because inflammation in the vagina does not occur
Vaginal discharge with a very fishy odor, especially fater sex
Itching is common
VAGINITIS - Infection of the vagina, with inflammation
Most common cause: Candida albicans – a yeast (fungus) – yeast infection
Other causes: bacterial, as in the case of Gardnerella; or even protozoal, as in the case of Trichomonas vaginalis.
Gram stain of sample of vaginal discharge
Note that there are 2 white blood cells (left bottom).
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

21. Staining Techniques: Clinical Applications (Chapter 3)
Microbiology
4/25/2017
Staining Techniques: Clinical Applications (Chapter 3)
A sputum sample from Calle, a 30-year-old Asian elephant, was smeared onto a slide and air dried. The smear was fixed, covered with carbolfuchsin, and heated for 5 minutes. After washing with water, acid alcohol was placed on the smear for 30 seconds. Finally, the smear was stained with methylene blue for 30 seconds, washed with water, and dried. On examination at 1000X, the zoo veterinarian saw red rods on the slide.
What infection do the results suggest? (Calle was treated and recovered.)
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

22. Staining Techniques: Clinical Applications (Chapter 3)
Microbiology
4/25/2017
Staining Techniques: Clinical Applications (Chapter 3)
The acid-fast stain binds strongly only to bacteria that have mycolic acid in their cell walls, a waxy material. This stain is used to identify all bacteria in the genus Mycobacterium, including Mycobacterium tuberculosis, the causative agent of tuberculosis, and Mycobacterium leprae, cause of leprosy. This stain is also used to identify the pathogenic strains of the genus Nocardia. Bacteria in the genera Mycobacterium and Nocardia are acid-fast.
ACID-FAST STAIN – Chapter 3 – Tortora, 10 Ed.
Figure 3.13 Acid-fast bacteria. The Mycobacterium leprae bacteria that have infected this tissue have been stained red with an acid-fast stain. Non–acid-fast cells are stained with the methylene blue counterstain.
Another important differential stain (one that differentiates bacteria into distinctive groups) is the acid-fast stain, which binds strongly only to bacteria that have a waxy material in their cell walls.Microbiologists use this stain to identify all bacteria in the genus Mycobacterium, including the two important pathogens Mycobacterium tuberculosis, the causative agent of tuberculosis, and Mycobacterium leprae (leprı), the causative agent of leprosy. This stain is also used to identify the pathogenic strains of the genus Nocardia (no -karde-a). Bacteria in the genera Mycobacterium and Nocardia are acid-fast.
In the acid-fast staining procedure, the red dye carbolfuchsin is applied to a fixed smear, and the slide is gently heated for several minutes. (Heating enhances penetration and retention of the dye.) Then the slide is cooled and washed with water. The smear is next treated with acid-alcohol, a decolorizer, which removes the red stain from bacteria that are not acid-fast. The acid-fast microorganisms retain the red color because the carbolfuchsin is more soluble in the cell wall lipids than in the acid-alcohol (Figure 3.13). In non–acid-fast bacteria, whose cell walls lack the lipid components, the carbolfuchsin is rapidly removed during decolorization, leaving the cells colorless. The smear is then stained with a methylene blue counterstain. Non–acid-fast cells appear blue after application of the counterstain.
CHAPTER 3 – COWAN’S BOOK
The acid-fast stain, like the Gram stain, is an important diagnostic stain that differentiates acid-fast bacteria (pink) from non-acid-fast bacteria (blue). This stain originated as a specific method to detect Mycobacterium tuberculosis in specimens. It was determined that these bacterial cells have a particularly impervious outer wall that holds fast (tightly or tenaciously) to the dye (carbol fuchsin) even when washed with a solution containing acid or acid alcohol. This stain is used for other medically important mycobacteria such as the Hansen’s disease (leprosy) bacillus and for Nocardia, an agent of lung or skin infections.
Figure 3.13 Acid-fast bacteria.
The Mycobacterium bacteria that have infected this tissue have been stained red with an acid-fast stain. Non–acid-fast cells are stained with the methylene blue counterstain.
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

23. Special Staining Techniques
Microbiology
4/25/2017
Special Staining Techniques
Negative staining – capsules: using India ink or nigrosin for background and safranin; capsules appear as halos surrounding each stained bacterial cell (red).
Endospore staining: Schaeffer- Fulton: malachite green and heat to help stain penetrate endospore; safranin for other cellular parts.
Flagella staining: carbolfuchsin and a mordant to make flagella wide enough to be seen.
QUESTION: Discuss 2 other examples of staining techniques, and when they are used.
SPECIAL STAINING TECHNIQUES
Negative Staining for Capsules
Many microorganisms contain a gelatinous covering called a capsule, which we will discuss in our examination of the prokaryotic cell in Chapter 4. In medical microbiology, demonstrating the presence of a capsule is a means of determining the organism’s virulence, the degree to which a pathogen can cause disease.
Capsule staining is more difficult than other types of staining procedures because capsular materials are soluble in water and may be dislodged or removed during rigorous washing. To demonstrate the presence of capsules, a microbiologist can mix the bacteria in a solution containing a fine colloidal suspension of colored particles (usually India ink or nigrosin) to provide a contrasting background and then stain the bacteria with a simple stain, such as safranin (Figure 3.14a). Because of their chemical composition, capsules do not accept most biological dyes, such as safranin, and thus appear as halos surrounding each stained bacterial cell.
Endospore (Spore) Staining
An endospore is a special resistant, dormant structure formed within a cell that protects a bacterium from adverse environmental conditions. Although endospores are relatively uncommon in bacterial cells, they can be formed by a few genera of bacteria. Endospores cannot be stained by ordinary methods, such as simple staining and Gram staining, because the dyes do not penetrate the wall of the endospore.
The most commonly used endospore stain is the Schaeffer- Fulton endospore stain (Figure 3.14b). Malachite green, the primary stain, is applied to a heat-fixed smear and heated to steaming for about 5 minutes. The heat helps the stain penetrate the endospore wall. Then the preparation is washed for about 30 seconds with water to remove the malachite green from all of the cells’ parts except the endospores. Next, safranin, a counterstain, is applied to the smear to stain portions of the cell other than endospores. In a properly prepared smear, the endospores appear green within red or pink cells. Because endospores are highly refractive, they can be detected under the light microscope when unstained, but they cannot be differentiated from inclusions of stored material without a special stain.
Flagella Staining
Bacterial flagella (singular: flagellum) are structures of locomotion too small to be seen with a light microscope without staining. A tedious and delicate staining procedure uses a mordant and the stain carbolfuchsin to build up the diameters of the flagella until they become visible under the light microscope (Figure 3.14c).Microbiologists use the number and arrangement of flagella as diagnostic aids. Animation Staining.
2011
Evelyn I. Milian - Instructor
Evelyn I. Milian - Instructor

24. SC300: Big Ideas in Science
4/25/2017
Questions???
INSTRUCTOR INFORMATION
Instructor and Credentials
Evelyn I. Milian, M.S.; Microbiology
Kaplan Address
AIM (AOL Instant Messenger) Name
milianevelyn
AIM Office Hours
By appointment. Contact me by or AIM at the above addresses to set one up.
Please keep your Course Syllabus handy and use it as your guide throughout the entire term.
Read all sections of the syllabus and make sure that you understand them.
Do not hesitate to contact me by or AIM as soon as you have a question.
* Students in other sections: Please contact your instructor for specific questions about assignments or grading.
2011
Evelyn I. Milian - Instructor
Prof. Evelyn I. Milian

25. Some Recommendations to be Successful in the Course
SC300: Big Ideas in Science
4/25/2017
Some Recommendations to be Successful in the Course
Read your s and course announcements every day.
Work consistently and stay caught up; make a plan and stay organized.
Post early and often to the Discussion Board.
Keep up with the assignments and projects.
Come prepared to the weekly seminars and participate meaningfully throughout the entire hour.
Always support all your work with complete APA style references and in-text citations and rely on quality literature resources.
Avoid copying and pasting anything longer than a line or two from any given source; posts and projects must always be made up of your own words, supported by literature sources properly cited.
Make sure that you meet the learning activity submission deadlines in each Unit. Units start on Wednesdays each week and end at 11:59 PM Eastern Time the following Tuesday.
Use all Kaplan resources available to help you in your courses.
2011
Evelyn I. Milian - Instructor
Prof. Evelyn I. Milian

26. Evelyn I. Milian - Instructor
Microbiology
2010
References
Alters, Sandra & Alters, Brian. (2006). Biology, Understanding Life. John Wiley & Sons, Inc. NJ, USA.
Audesirk, Teresa; Audesirk, Gerald & Byers, Bruce E. (2005). Biology: Life on Earth. Seventh Edition. Pearson Education, Inc.-Prentice Hall. NJ, USA.
Black, Jacquelyn G. (2005). Microbiology, Principles and Explorations. Sixth Edition. John Wiley & Sons, Inc. NJ, USA.
Campbell, Neil A.; Reece, Jane B., et al. (2008). Biology. Eighth Edition. Pearson Education, Inc.-Pearson Benjamin Cummings. CA, USA.
Cowan, Marjorie Kelly; Talaro, Kathleen Park. (2009). Microbiology A Systems Approach. Second Edition. The McGraw-Hill Companies, Inc. NY, USA.
Dennis Kunkel Microscopy, Inc. (2010).
Mader, Sylvia S. (2010). Biology. Tenth Edition. The McGraw-Hill Companies, Inc. NY, USA.
Tortora, Gerard J.; Funke, Berdell R.; Case, Christine L. (2010). Microbiology An Introduction. Tenth Edition. Pearson Education, Inc.-Benjamin Cummings; CA, USA.
OTHER REFERENCES:
Alters, Sandra & Alters, Brian. (2006). Biology, Understanding Life. John Wiley & Sons, Inc. NJ, USA.
Belk, Colleen; Borden, Virginia. (2007). Biology, Science for Life. Second Edition. Pearson Education, Inc- Prentice Hall. NJ, USA.
Brooker, Robert J.; Widmaier, Eric P.; Graham, Linda E.; Stiling, Peter D. (2008). Biology. The McGraw-Hill Companies, Inc. NY, USA.
Dennis Kunkel Microscopy, Inc. (2010).
Mader, Sylvia S. (2007). Essentials of Biology. The McGraw-Hill Companies, Inc. NY, USA.
Marieb, Elaine N. (2006). Essentials of Human Anatomy & Physiology. Eighth Edition. Pearson Education, Inc., publishing as Benjamin Cummings. CA, USA.
Presson, Joelle & Jenner, Jan. (2008). Biology, Dimensions of Life. The McGraw-Hill Companies, Inc. NY, USA.
Solomon, Eldra; Berg, Linda; Martin, Diana W. (2008). Biology. Eighth Edition. Cengage Learning. OH, USA.
Tortora, Gerard J.; Derrickson, Bryan. (2006). Principles of Anatomy and Physiology. Eleventh Edition. John Wiley & Sons, Inc. NJ, USA.
Tortora, Gerard J.; Funke, Berdell R.; Case, Christine L. (2007). Microbiology An Introduction. Ninth Edition. Pearson Education, Inc.-Benjamin Cummings; CA, USA.
OLD REFERENCES:
Atlas, Ronald M. & Bartha, Richard. (1998). Microbial Ecology – Fundamentals and Applications. Fourth Edition. Benjamin/Cummings Publishing Company, Inc. CA, USA.
Benson, Harold J. (2002). Microbiological Applications: Laboratory Manual in General Microbiology. Eighth Edition. The McGraw-Hill Companies; NY, USA.
Maier, Raina M.; Pepper, Ian L. & Gerba, Charles P. (2000). Environmental Microbiology. Academic Press- Elsevier. CA, USA.
Prescott, Lansing M.; Harley, John P.; Klein, Donald A. (1996). Microbiology. Third Edition. Wm. C. Brown Publishers; IA, USA.
2011
Evelyn I. Milian - Instructor
Prof. Evelyn I. Milian