Microbial Control

Outside the Body

Physical Methods

I.  Heat
 Generally elevated temperatures are microbicidal and lower temperatures are microbistatic
 Can be moist (steam) or dry heat (changing temperature of air)

1. Moist Heat
 Pressurized Steam
o Pressure raises the temperature of steam
o Most efficient pressure-temperature combination for sterilization--kills spores as well
 Autoclave is an example
 Pasturization
o Used to disinfect beverages
 Does not kill endospores or thermoduric microbes
o Heat is applied to liquids to kill potential agents of infection, while retaining the liquid's flavor and
food value
 Boiling Water
o For disinfection
2. Dry Heat
 Incineration
o Ignites and reduces microbes to ashes and gas
o Common practice in microbiology lab—bacincinerators
o Sterilization
  Dry Oven
o Used for heat-resistant items that do not sterilize well with moist heat such as electronic
equipment
o Usually an electric oven
o Sterilization
3. Cold
 Slows down the growth of cultures and microbes in food during processing and storage
 Does not kill microbes, only preserves them (microbistatic)
 Desiccation is the dehydration of vegetative cells that might direct the cell into an endospore state
 Lyophilization is a combination of freezing and drying that is used to preserve microorganisms in a viable
state for many years 

II. Radiation
 Energy emitted from unstable atoms that are used for microbial control
o Gamma rays, X rays, UV 
 Usually disinfection (nonionizing UV) rather than sterilization (ionizing xray, gamma ray)
 Hospital rooms, operating rooms, schools, food
 Drinking water or purifying liquids

Chemical Methods

 Occur in liquid, gaseous, or solid state

 Tinctures are solutions dissolved in pure alcohol or water-alcohol mixtures

1. Halogen Antimicrobial Agents

 F, Cl, Br, I
 Microbicidal and sporicidal with longer exposure
 Chlorine compounds can be liquid, gaseous, hypochlorites, or chloramines
o Kills bacteria, endospores, fungi and viruses
o Household bleach
 Iodine compounds--free iodine and iodophors (diluted with water)
o Can be topical antiseptic and/or disinfectant
o Betadine is the most common formulation
2. Phenol and its derivatives
 Great against gram positives because they denature proteins, disrupt lipids and enzyme function
 At high concentrations they are poisonous for cells because:
o Disrupt cell membranes
o Percipitate proteins
 At lower concentrations they inactivate certain critical enzyme systems
 Microbicidal against vegetative microbes including viruses
o NOT serilizing
 At concentrations below 1% they are used as throat lozenges due to tis anesthetic properties
 General disinfectants include
o Lyson
o Triclosan 
 Bisphenol that is a popular component of antibacterial soaps.
 Inhibits an enzyme needed for the synthesis of fatty acids
 Effective against gram negatives and yeast
 Phenolics are derivatives that have been modified to reduce the irritating properties and increase
antimicrobial activity in the presence of soaps or detergents
3. Chlorhexidine
 Complex base containing chlorine and two phenolic rings (biguanide)
 Targets cell membranes and protein structure
  At moderate to high concentrations it is bactericidal for both gram positve and gram negative
 Inactive against spores
 Mild, low toxicity and rapid action
 Disinfecting
 When combined with a detergent it is used as a surgical scrub
4. Alcohols 
 Kills bacteria and fungi but not endospores and nonenveloped viruses
 Mechanism of action depends on its concentration
o 50% and higher dissolve membrane lipids, disrupt cell surface tension and compromise membrane
integrity
o 50% - 90% denature proteins through coagulation
o 100% dehydrates cells and inhibits growth
 Ethyl and isopropyl alcohols
 Not good for degerming wounds because they cause coagulation under which bacteria still grow 
 Can be used for antisepsis
5. Hydrogen Peroxide and Ozone
 Germicidal effects due to the direct and indirect actions of oxygen
 Bactericidal, viricidal and fungicidal
 In higher concentrations it is sporicidal
 Can be sterilizing as in contact lens solutions
 Ozone has similar effects to hydrogen peroxide
o used to disinfect water
6. Heavy Metal Compounds
 Hg, Ag, Au, Cu, As, Zn
o Hg and Ag are germicides
 Have oligodynamic action
o have antimicrobial effects in small amounts
o Bind onto functional groups of proteins and inactivate them
7. Detergents
 Act as surfactants
 Cationic detergents are more effective because the positively charged end binds well with the
predominantly negatively charged bacteria surface proteins
 Weak microbicides but gain germicidal value when mixed with chlorohxidine or iodine
8. Aldehydes
 Germicides
 Gluteraldehyde
o Liquid
o Cross-links proteins on cell surface
o Inactivates enzymes within cells
o Does not destroy endospores
o Toxic to humans
 Formaldehyde
o Gas--can be used to disinfect a whole room
o Toxic to humans
o can be used as a perservative of tissue samples at 40% and higher concentrations
9. Gaeous mixtures
 Chlorine dioxide and ethylene dioxide are sterilants
10. Acids and Alkalis
 Low or high pH can inhibit microbial cells
 Limited in application due to corrosive nature
 Often used as a preservative in food
o benzoic , Lactic acids
 Mechanical Methods

1.  Applications of Filtration
  Prepare liquids that can’t withstand heat—can decontaminate beverages without altering their flavor—
antibiotics can go through an IV
 Water purification
   Removing airborne contaminants (HEPA filters)

Inside The Body

Antibacterials
Target: Cell Wall--Mechanism: Blocks Synthesis and Repair

I. Beta-Lactam ring 
  1. Penicillin--extracted from the fungi Penicillium Notatum
  Consists of three basic parts—a thiazolidine ring, a beta-lactam ring, and a variable side which dictates its
anti-microbial activity
 Affects gram positive bacteria
 Bactericidal 
 Generally well tolerated and has low toxicity to host
 Mode of Action: Disrupts the formation of the cell wall by inhibiting enzymes essential for peptidoglycan
synthesis, which causes the cell wall to have weak points, leading to lyses
 o Specifically binds to the enzyme that is responsible for cross linking glucosamine and muramic
acid.
 The only requirement for them to work is that the cell must be actively growing and therefore making
peptidoglycan (exponential phase)
o Mycobaacterium, though it is a gram positive bacteria is not damaged by penicillin group because
it is not very active—its growth phase is very long and it contains Myacolic acids.
 Penicillinase and/or beta-lactamase are enzymes produced by bacteria which destroy the beta-lacatam ring
of the penicillin molecule, and therefore deactivate the drug
 Penicillin Group
o G and V
 Most important, natural forms
o Ampicillin, carbenicillin and amoxicillin
 Semi-Synthetic
 Broader spectrum and more stable then first generation
o Methicillin, nafcillin, and cloxacillin
   Penicillinase-resistant penicillins
o Mezlocillin and azlocillin
 Extended spectrum and can be sutsituted for combinations of antibiotics
o Combinations—used for systemic infections, contain compounds which inhibit beta-lactamase and
are used for systemic infections
 Augmentin
 Combination called Clavamox: amoxicillin and clavulanic acid
 Claculanic acid inhibits beta-lactamase enzymes and longevity
  Zosyn
 Combination of Tazobactam and piperacillin
 Tazobactam inhibits beta-lactamase emzymes
  2. Cephalosporin--extracted from the fungus cephalosporium
 Have beta-lactam ring structure that can be synthetically altered and so has similar mode of action to
penicillin and is resistant to most penicilinases
 Mode of Action: Bind to penicillin-binding proteins that interfere with the synthesis of peptidoglycan
 Relatively broad-spectrum
 Cause fewer allergic reactions then penicllins
 3. Monobactams
 Aztreonam narrow spectrum drug—aerobic gram negatives that is useful for treating penicillin-allergic
patients because of the different chemical structure and so it is resistant to beta-lactamases
 Binds primarily to one of the penicillin-binding proteins in gram-negative aerobe
 4. Carbapenems 
 Widest spectrum of antibacterial activity
 Binds to penicillin-binding proteins and cause cell elongation and lysis
 Stable against beta-lactamase
 Bacteria produce resistance by the production of carbapenemase and can alter the porin channels, which
reduces the permeability of the drugs

II. Non-Beta Lactam Ring

    1. Bacitracin--derived from bacterium Bacillus
 Often combined with polymyxins in topical application
 Active against gram-positive bacteria
 Very toxic to Kidneys
o Bacitracin is administered only topically
 Mode of Action: Inhibits bacterial cell wall synthesis and disrupts cell membrane
    2. Fosfomycin
 Used as a single-dose therapy for UTI's
o Multi-use can cause bacterial resistance to develop quickly
 Broad-spectrum of activity
  Mode of Action: Inhibits pyruvyl transferase which is necessary in the first steps of peptidoglycan synthesis
o Bacteriacidal
    3. Vancomycin--produced by streptomyces
 Active against aerobic and anaerobic gram positive organisms including Methicillin Resistant staplococci
(MRSA)
o Not active against gram negative or mycobacteria because of the molecule's large size
 Can't penetrate the outer membrane
 Mode of Action: Inhibits peptidoglycan synthesis in the cell wall
o Bactericidal 

Target: Protein Synthesis Inhibitors

I. 30S Ribosomal Subunit Inhibitors (-mi(y)cin and -cyclins)

    1. Aminocyclitols 
 Mode of Action: Binds reversibly to the 30S ribosomal subunit and inhibits protein synthesis
o Bacteriostatic
 Has a very limited spectrum--specifically used to treat Neisseria gonorrhoeae
    2. Aminoglycosides
 Mode of Action: Binds irreversibly to the bacterial 30S subunit and causes misreading of the genetic code
that results in the production of nonsense proteins
o Bactericidal 
 Medications include those that end in -mycin
o Streptomycin, Neomycin, Kanaycin, Tobramycin, and Gentamicin 
 Has considerable toxicity
o Nephrotoxicity and Auditory toxicity 
o Neomycin can't be administered via IV because it is so toxic--mainly for topical treatment and
rarely oral
    3.  Tetracyclines
 Mode of action: Binds reversibly to the 30S ribosomal subunit 
o Bactericidal
 Broad spectrum
 Administered orally
 Medications include
o Doxycycline (used to treat anthrax), Minocycline and Tetracycline (blocks tRNA docking site)
    4. Glycylcyclines
 Synthetic derivatives of tetracyclines
 Mode of Action: Binds reversibly to the 30S subunit
o Bacteriastatic
 Tigecycline is only approved drug so far 
 Resistant to most resistant mechanisms that effect tetracyclines
II. 50S Ribosomal Subunit inhibitors
    1. Macrolides (-ithromycin)
 Mode of Action: Inhibits bacterial ribosomal RNA 
o bacteriostatic
 Bactericidal against streptococci
o strep. pyogenus (strep throat)
 Broad spectrum 
 Medications include
o Azithromycin, Clarithromycin 
 Legionella, Mycoplasma
o Dirithromycin, 
o Erythromycin is not broad spectrum because it can't penetrate gram negative walls
    2. Ketolides
  Mode of Action: Inhibits the translation at the level of the 50S ribosomal subunit
 Medication includes Telithromycin a derivative of Erythromycin
 Spectrum of Activity
o Good against respiratory pathogens
 Haemophilus influenzae, Streptococcus pneumoniae
o Good against intracellular bacteria
 Legionella, Mycoplasma, Chlamydia
o Designed specifically for the treatment of community aquired respiratory tract infections
    3. Lincosamides
 Mode of Action: Binds to the 40S ribosomal subunit
 Can be bactericidal or static depending on drug concentration, bacterial species, and bacterial density
 Broad Spectrum
 Medications include
o Clindamycin and Lincomycin
 Clindamycin has better antibacterial activity and improved absorption after oral
administration
    4. Oxazolidones
 Mode of Action: Binds to the 50S subunit, inhibits the formation of a functional 70S complex that prevents
the initiation of translation
 Medication Include Linezolid
 Spectrum of Activity
o Generally bacteriostatic
o Bactericidal against most strains of pneumococci
o Used for MRSA and vancomycin-resistant enterococci
    5. Phenicols
 Mode of Action: Inhibits protein synthesis by binding reversibly to the 50S ribosome
o Bacteriostatic
 Medication Includes Chloramphenicol
 Has a broad spectrum of activity
 Limited to seriously ill patients due to the side effect that causes irreversible damage to the bone marrow
which leads to aplastic anemia
    6. Streptogramins
 Mode of Action: Inhibits bacterial protein synthesis, binds irreversibly to the 50S subunit
o Bactericidal
 Medication Include a combination drug called Synercid made up of Quinupristin-Dalfopristin (30:70)
 Used to treat seriously ill or life threatening infections accociated with vancomycin-resistant Enterococcus
faecium (VREF)

Target: DNA & RNA

    1. Rifampin
 Related to the Macrolides
o Medication is Ansamycin
 Limited spectrum because molecule can't pass through the cell envelope of many gram-negative bacilli
 Mode of Action: Forms a stable complex with RNA polymerase preventing transcription
 Rifampin-resistant bacteria possesses an altered RNA polymerase that comes from mutations during
monotherapy with Rifampin
o Important when treating mycobacteria
 A side effect is that all secretions become orange
    2. Quinolones and Fluroquinolones
  Mode of Action: Targets DNA gyrase, an enzyme essential for replication, recimbination and repair.
Inhibition of these enzymes causes the relaxation of the supercoiled DNA
o This terminates chromosomal replication and interferes with cell division and gene expression
o bactericidal
 Fluroquinolones are synthetics that have the addition of a flourine
o Have the suffix –floxacin
Quinolones and Fluroquinolones
Subclass Generic Names
Quinolone (1st generation) Narrow
Cinoxacin, Niladixic acid
spectrum active against gram
(UTIs)
positive cocci
Fluroquinolone (2nd generation) Ciprofloxacin(anthrax),
Broad-spectrum against both gram Enoxacin, Fleroxacin,
positive and gram negative. Less Levofloxacin, Lomefloxacin,
toxic than first generation Norfloxacin, Ofloxacin
Fluroquinolone (3rd generation)
Increased activity against gram- Sparfloxacin
positive cocci
Fluroquinoline (4th generation)
Gatifloxacin, Gemifloxacin,
Increased activity against gram-
Moxifloxacin, Trovafloxacin
positive cocci

    3. Metronidazole (Flagyl)

 A type of prodrug that must first be metabolized by the body before it can be used
 Mode of Action: When the nitro group enteres the cell, it is reduced in the cytoplasm and forms cytotoxic
compounds that disrupt the bacterial or protozoan DNA
o Bactericidal 
o Only against obligate anaerobes like Clostridum
 Medications include
o Nitrazoximide has anti-helminthic activity
o Fasigyn

Target: Cell Membrane

    1. Polypeptides--Polymyxin B
 Mode of Action: Interact with the phospholipids in the cell membrane and increase the cell's permeability
o Bactericidal
 Very toxic to the kidneys and neurotoxic 
 Often combined with bacitracin (attacks the cell wall) in topical applications
o Polymyxin B is active against gram-negative bacilli like Psudomonas and Bacitracin is active
against gram-positive bacteria
 Together have broad spectrum of activity
 Currently being used to treat  Acinetobacter baumannii, which is multi-drug resistant (MDRAB)
    2. Lipopeptides
 Mode of Action: concentration-dependent bactericidal activity
o Causes depolarization of the cell membrane
 Medications include Daptomycin (cubicin) a European medication
 Spectrum of Activity
o Gram positive organisms including methicillin-susceptible and resistant Staphylococci,
Streptococci, Enterococci, and Bacillus
 o Not active against gram-negative organisms or mycobacteria
o Can work synergistically with aminoglycosides or rifampin
 This allows you to lower the concentration of the other medications

Target: Folic Pathway Inhibitors

 Medications include
o Sulfonamides, Trimethoprim (TMP)
o Trimethoprim-sulfamethoxazole  (TMP-SMX; trade names Bactrim or Septra)
 Mode of Action: Block the bacterial folic acid metabolic pathway at different sites
 TMP-SMX acts synergisticallyd
 First synthetic antibiotics 

Antifungals
Because fungal cells are eukaryotic, anti-fungals can also be toxic to human cells

1. Macrolide polyenes (antibiotics)

 Mode of Action: Bind to fungal cells and cause loss of selective permiability
o Binds to a sterol in the fungal cell membranes called ergosterol which human cells do not contain;
we contain cholesterol
 Medications Include
o Amphotercin B--most common; used to treat streptomyces 
o Nystatin
 Used to treat candidias (yeast infections) of the skin and mucous membrane
 Developed at Albany, NY
 Has nephrotoxicity 
2. Synthetic Azoles
 Mode of Action: Inhibits ergosterol synthesis which weakens the fungal cell membrane
 Broad-spectrum
 Medications include
o Clotrimazole
o Fluconazole
o Miconazole
o Ketoconazole
 Newer, broader spectrum, orally

Antiparasitics
Drugs used to treat protozoan and helminth infections
1. Quinine
 Extracted from the bark of the cinchona tree
 Was the principle treatment for malaria for hundereds of years
o Replaced by the synthetic quinolines
 NOT TO BE CONFUSED WITH QUINOLONES
o Chloroquine, Primaquine, Mefloquine (has psychiatric side effects)
2. Metronidazole
 Used to treat mild to severe intestinal infections and hepatic disease caused by Entameoba histolytica
 Can also be used to treat Giardia lamblia and Trichomonas vaginalis
 Metabolized
 3. The antibacterial groups of sulfonamides and tetracyclines also have antiprotozoan activities 

Antivirals
1. Inhibition of Virus Entry
 Receptor binding
 Fusion
o Fuzeon, enfuviritide, very costly and mimics region on virus that is responsible for fsion
 Uncoating
o Amantide (H3N2, H1N1, Avian, Influenza B are now all resistant)
o Tamiflu, Relenza
2. Inhibition of Nucleic Acid Synthesis
 Acyclovir--herpes
 Nucleotide analog reverse transcriptase (RT) inhibitors--retroviral, specifically HIT
o Zidovudine (AZT)
 Non-nucleoside reverse transcriptase inhibitors
o Nevirapine 
3. Inhibition of Viral Assembly/Release
 Protease inhibitors 
o Saquinavir
o Amantidine
 
Interferon, a glycoprotein produced by fibroblasts and leukocytes in response to immune stimuli can also be used to
treat viral infections. It has anti-viral and anti-cancer properties and plays a major role in natural immunities 

Antiviral Intended Virus

Amantadine & Rimantadine
uncoating
Zanamivir & Oseltamivir NA
Acyclovir & Famciclovir
replication
Influenza A
Influenza A & B
Herpes simplex virus (HSV)
Varicella-zoster virus (VZV)
(chickenpox, shingles)
Ribavirin RNA
Ganciclovir & Foscarnet
nucleotide analogs
Nucleoside reverse transcriptase
inhibitors (i.e. Zidovudinez (AZT)
and Didanosine)
Non-nucleoside reverse
transcriptase inhibitors (i.e.
Nevirapine and Delavirdine)
Protease inhibitors (i.e. Ritonavir
and Saquinavir)
Lamivudine nucleocide analog
Interferon-α
Respiratory syncytial virus,
hepatitis C virus, others
Cytomegalovirus (CMV)
HIV Human immunodeficiency
virus
HIV
HIV
Hepatitis B
Hepatitis B and hepatitis C