Overview for this class:

Plan of action:
Review from last class
QUIZ 2 Due today at 5pm on Sakai site

Lecture on Bacteria
Robert Koch father of microbiology Define cell shape and structure Contrasts modes of bacterial motion Bacterial growth and division Contrast oddball bacteria with normal bacteria

Preview for Virus

Kochs Postulates

http://www.daviddarling.info/images/Koch_Robert.jpg

Kochs Postulates

- Association - Isolation - Causation - Re-Isolation

http://www.daviddarling.info/images/Koch_Robert.jpg

Cell Shapes and Patterns

Morphology: bacteria are found in several common shapes (and arrangements), which are useful in species identification
Bacillus, (bacilli, pl.) rod shaped Coccus, (cocci, pl.) spherical Curved or Spiral, (Vibrio, curved, Spirilla, rigid helix or wavy, and
Spirochete, flexible helix)

Figure 01: Bacterial cell shapes and patterns

Naming Bacteria
Bacteria are named using Linnaeuss binomial classification system (having two Latinized names)
genus name, capitalized (Escherichia) species name, not capitalized (coli) both names are italicized: Escherichia coli

Bacteria names are frequently instructive Escherichia is named for Theodor Escherich coli indicates its habitat (large intestine) Streptococcus indicates shape and arrangement

Anatomy of the Bacterial Cell

Figure 02: A composite bacterial cell

Envelopeis external to the cytoplasm

Plasma membrane, selectively permeable Cell wall (gram positive vs. gram negative)
G+: thick layer of rigid polymer, peptidoglycan G: thin layer of peptidoglycan Prevents osmotic rupture of cell membrane

Outer membrane (in gram negatives only)

Contains fever inducing endotoxin

Capsule not integral to the cell

Is a virulence factor, anti-phagocytic

The Gram stain is an important first step in the identification of bacteria pathogens.
Choice of antibiotics is influenced by the Gram stain reaction
Broad spectrum antibiotics work against g+ and g Narrow spectrum antibiotics work against g+ or g-

Figure 03: Gram Stain

Cytoplasmis all of a cells contents enclosed within the plasma membrane

Nucleoid, region of cytoplasm containing chromosomal DNA
Double stranded DNA (dsDNA) One or more circular and/or linear chromosomes E. coli, one circular Vibrio cholerae, 2 circular Borrelia spp., linear and circular

Figure 05: Binary fission

DNA super coiling and blown up DNA

Plasmids, small, circular, independently replicating, dsDNA, Encode limited number of genes (few to many) Expand genetic capability of host cell

http://cmgm.stanford.edu/biochem201/Slides/DNA%20Topology/072%20Supercoiled%20DNA%20jpeg.JPG

Cytoplasm (cont.)
Spores (endospores, formed within the cell)
viable for long periods (perhaps centuries or longer) resistant to heat, boiling, drying, radiation, and various chemical compounds, including alcohol Important pathogens Bacillus anthracis, anthrax, possible bioweapon Clostridium spp., tetanus, botulism, gas gangrene (all anaerobes)
Figure 07: Bacillus anthracis
Courtesy of Dr. Sherif Zaki and Elizabeth White/CDC

The spore cycle: vegetative cells sporulate to produce spores, and spores germinate into vegetative cells.

Figure 08: The spore cycle

Bacterial Growth
Limits to growth/multiplication of bacteria
Abiotic, temperature, the availability of oxygen and water, etc. Biotic, disease, competition, and predation

Figure 10: Overnight growth in a broth tubeclear to turbid

Examination of the dynamics of growth reveals the presence of four major growth phases.
The Four Growth Phases
Lag, period of adaptation to new conditions Exponential/logarithmic, the cell population doubles with each generation (23 = 2 x 2 x 2 = 8; the exponent 3 equals the number of generations) Stationary, rate of cell division is about equal to the rate of death (nutrients are depleted and toxins accumulate) Death, the number of cells dying exceeds the rate of cell division.

A typical growth curve for E. coli

Figure 11: Growth curve of E. coli

Significance of Bacterial Growth

Growth during an infection mirrors the 4 stages Lag, a few bacteria try and establish themselves at site of infection Log, numbers of bacteria rising exponentially, see first signs and symptoms for that infection Stationary, precipitated by success of host immune system, or success of antibiotic treatment Death, bacterial numbers drop precipitously, due to effects of immune system or antibiotics

Me

You

Time to get interactive

http://www.hhmi.org/biointeractive/media/b acterial_growth-lg.wmv

At what time would we realize that resources are running out?

100%

Percent of container filled

90% 80% 70% 60% 50% 40% 30% 20% 10% 0% 24.00 23.59 23.58 23.57 23.55 23.50
Empty Filled

Time

Appendages
Flagella, used for motility
Rotates like propeller Chemotaxis
Move toward attractant Move away from repellant

Long, hollow, filament made of subunits of flagellin Many arrangements (single,

polar, bipolar, dispersed, etc.)
Figure 09: Structure and arrangement of flagella

Appendages (cont.)
Pili, (sing. pilus)
Shorter, straighter, thinner than flagella Filaments made of subunits of pilin protein Function
Adhesin, anchor for colonization of host cells and other surfaces pili, forms a bridge between donor and recipient bacterial cells, for transmission of DNA

http://www.youtube.com/watch?v=6p9e0oo lbmE&feature=BFa&list=PL1BC51C672B2 2EDD2&lf=results_video

Culturing Bacteria: Diagnostics

A clinical specimen is obtained to grow in culture to identify the suspected cause of the infection
throat swab, urine or blood culture, etc.

Inoculated into growth medium Streaked across agar plate media to aid identification Colonies may have identifiable morphology or properties Characteristic, texture, size, pigment, hemolysis, etc.

Courtesy of Robert Krasner Courtesy of Dr. Richard Facklam/CDC

Culturing Bacteria: Diagnostics (cont.)

A variety of media and diagnostic tests are available Type selected depends on source of the specimen (bacterial pathogens of the skin may differ from those typically found in a vaginal swab, etc.) Some bacteria cant be grown, or they grow too slowly, requiring alternative approaches Detect specific anti-bacterial antibodies in patients blood Amplification of pathogens DNA

Oddball (Atypical) Bacteria

Mycoplasmas
Have no cell wall, thus, have no shape Very small and require specialized media for growth Disease: walking pneumonia

Chlamydiae
Obligate intracellular parasites Disease: urethritis, trachoma, lymphogranuloma venereum

Rickettsiae
Obligate intracellular parasites Transmitted by arthropods (except for Q fever)

Overview for this class:

Plan of action:
Review from last class Lecture on Virus
Virus as infectious disease - historical Discovery of viral particles Viral structure and replication Culture of virus

Preview for Bacterial Genetics 1