CONTROLLING THE GROWTH OF MICROBES

DEFINITION OF TERMS
Before discussing the various methods used to destroy or inhibit microbes, a number of terms should
be understood as they apply to microbiology.
Sterilization
The complete destruction of all living organisms, including cells, viable spores, and viruses, is called
sterilization. When something is sterile, it is devoid of microbial life. Sterilization of objects can be
accomplished by heat, autoclaving (steam under pressure), gas (ethylene oxide), various chemicals
(such as formaldehyde), and certain types of radiation (e.g., ultraviolet light and gamma rays). These
procedures arc discussed later in this chapter.
Disinfection
Disinfection is the destruction or removal of pathogens from nonliving objects by physical or
chemical methods. The heating process developed by Pasteur to disinfect beer and wines is called
paste14rization. It is still used to eliminate pathogenic microorganisms from milk and beer. It should
be remembered that pasteurization is not a sterilization procedure, because not all the microbes are
destroyed. Chemical agents are also used to eliminate pathogens. Chemicals used to disinfect inanimate objects, such as bedside equipment and operating rooms, are called disinfectants. Disinfectants
are strong chemical substances that cannot be used on living tissue. Antiseptics are solutions used to
disinfect skin and other living tissues. Sanitization is the reduction of microbial populations to levels
considered safe by public health standards, such as those applied to restaurants.
Microbicidal Agents
The suffix "-cide" or "-cidal" refers to "killing," as in the words homicide and suicide. General terms
like germicidal agents (germicides), biocidal agents (biocides), and microbicidal agents
(microbicides) are disinfectants that kill microbes; such agents might be used in sanitization
procedures. Bactericidal agents (bactericides) are disinfectants that kill bacteria specifically, but not
necessarily bacterial endospores. Because spore coats are thick and resistant to the effects of many
disinfectants, sporicidal agents are required to kill bacterial endospores. Fungicidal agents
((fungicides) kill fungi, including fungal spores. Algicidal agents (algicides) are used to kill algae in
swimming pools and hot tubs. Viricidal (or virucidal) agents destroy viruses. Pseudomonicidal agents
kill Pseudomonas species and tuberculocidal agents kill Mycobacterium tuberculosis.
The mechanism by which various biocides kill cells varies from one disinfectant to another.
Some disinfectants target and destroy cell walls, whereas others attack cell membranes. Others
destroy enzymes or structural proteins or nucleic acids. Various factors affect the effectiveness of a
disinfectant and must be taken into consideration whenever a disinfectant is used. These factors
include the concentration of the disinfectant, the amount of time the disinfectant remains in contact
with the organisms, temperature, pH, and the presence of organic materials (such as blood, pus, etc.).

Microbistatic Agents
A microbistatic agent is a drug or chemical that inhibits growth and reproduction of microorganisms,
whereas a bacteriostatic agent is one that specifically~ly inhibits the metabolism and reproduction of
bacteria. Some of the drugs used to treat bacterial diseases are bacteriostatic, whereas others are
bactericidal. chapter). Freeze-dr~ng (lyophilizafl~on) and rapid freezing (using liquid nitrogen) are
microbistatic techniques that are uscd to prese~~ microbes for ruture use or studv.

Asepsis
Sepsis refers to the growth of pathogens on living tissues, whereas asepsis means the absence of
pathogens on h.ving tissues. Aseptic technique is designed to eliminate and exclude all pathogens by
sterilization of equipment, disinfection of the environment, and cleansing of bodv tissues with
antiseptics. Antiseptic technique, developed by Lister in 1867, is a V\~e of aseptic technique. Lister
used dilute carbolic acid (phenol) to cleanse surgical wounds and equipment and a carbolic acid

aerosol to prevent harmfil microorganisms from entering the surgical field or contaminating the
patient. Antisepsis is the prevention of infection.
Sterile Technique
When it is necessa~ to prevent all microorganisms from gaining entrance into a laborato~ or on to a
surgical field, sterile technique is followed.
FACTORS INFLUENCING MICROBIAL GROWTH
There are many environmental factors that enhance or inhibit the growth of microorganisms,
including temperature, moisture, osmotic pressure, pH, barometric pressure, gases, radiation,
chemicals, and the prescnce of neighboring microbes. Many concepts involving these factors may be
applied to our eve~~day lives and to laborator~ and hospital environments.
Temperature
For ever~ microorganism, there is an optimal temperature at which the organism grows best, a
minimal temperature below which it ceases to grow, and a maximal temperature above which it is
destroved. These temperature ranges differ greatly among organisms. In general, rate of growth and
metabolism are slower at low temperatures and faster at higher temperatures. The effect of
temperature changes vanes from one species to another.
Microorganisms that grow best at high temperatures are called thermophiles. These heat-loving
microbes may be found in hot springs, compost pits, silage, and near hydrothermal vents at the bottom
of the ocean. Thenuophilic cyanobacteria,
Physical Antimicrobial Methods
The physical methods commonlv used in hospitals, clinics, and laboratories to destroy or control
pathogens are heat, pressure, desiccation, radiation, sonic disruption, and filtration.
HEAT
Heat is the most practical, efficient, and inexpensive method of disinfection and sterilization of those
inanimate objects and materials that can withstand high temperatures. Because of these advantages, it
is the means most frequently employed.
Two factors, temperature and time, determine the effectiveness of heat for sterilization. There is
considerable vanation from organism to organism in susceptibilityT to heat; pathogens usua1~ are
more susceptible than nonpathogens. A]so, the higher the temperature, the shorter the time required to
lull the organisms. The thermal death point (TDP) of any particular species of microorganism is the
lowest temperature that will kill all the organisms in a standardized pure culture within a specified
period of time. The thermal death time (TDT) is the length of time necessar~T to sterilize a pure
culture at a specified temperature.
In practical applications of heat for sterilization, one must consider the material in which a
mixture of organisms and their spores may be found. Pus, feces, vomitus, mucus, and blood contain
proteins that sen~ as a protective coating to insulate the pathogens; when these substances are present
on bedding, bandagc5, surgical instruments, and syringes, very high heat is required to destroy
vegetative
(growing) microorganisms and spores. In practice, the most effective procedure is to wash away the protein debris
with strong soap, hot water, and a disinfectant, and then sterilize the equipment with heat.
Heat applied in the presence of moisture, as boiling or steaming, is more effective than dry
heat because moist heat causes proteins to coagulate. Because cellular enzymes are proteins, they
are also inactivated. This is exactly what happens when an egg is hard-boiled: the combination of
heat and moisture causes the proteins to coagulate. Moist heat sterilization is faster than dry heat
sterilization and can be done at a lower temperature; thus, it is less destructive to many materials
that othenvise would be damaged by higher temperatures.
The vegetative forms of most pathogens are quite easily destroyed by boiling; however,
bacterial endospores are particularly resistant to heat and drying. The autoclave, which combines
heat and pressure, offers the most effective yet inexpensive means of destroying spores. Two
examples of sporeformers are Clostridium tetani, the causative agent of tetanus, and C. botulinum,

which causes a severe form of food poisoning; their spores are usually found in contaminated dirt
and dust. Botulism food poisoning is preventable by properly washing and pressure cooking (autoclaving) the food.
Certain viruses are remarkably resistant to heat. A case in point is the hepatitis virus, which is
frequently transferred from one person to another by the reuse of contaminated syringes and
needles that have not been adequately sterilized. It is recommended that all equipment used in the
transfer of blood be sterilized in an autoclave at 1210C for 20 minutes or boiled for 30 minutes or
baked in an oven at 1800C for 1 hour.
Dry Heat.
Dry heat baking in a thermostatically controlled oven provides effective sterilization of metals, glassware, some
powders, oils, and waxes. These items must be baked at 1600 to 1650C for 2 hours or at 1700 to 1800C for 1 hour.
An ordinary oven of the type found in most homes may be used if the temperature remalns constant. The
effectiveness of dry heat sterilization depends on how deeply the heat penetrates throughout the material, and the
items to be baked must be placed so that the hot air circulates freely among them.
Incineration, or burning, is an effective means of destroying contaminated disposable
materials. An incinerator must never be overloaded with moist or protein-laden materials, such as
feces, vomitus, or pus, because the contaminating microorganisms within these moist substances
may not be destroyed if the heat does not readily penetrate and burn them. Flaming the surface of
heat-resistant material is an effective way to kill microorganisms on forceps and bacteriological
loops and, for many years, was a common laboratory procedure. Flaming is accomplished by holding the end of the loop or forceps in the yellow portion of a gas flame (Fig. 7-3). Open flames are
dangerous and, therefore, rarely used in modern laboratories, where either sterile, disposable,
plastic inoculating loops are used or heat sterilization of wire inoculating loops is accomplished
using electrical hcating devices (Fig 7-3).
Moist Heat.
As previously stated, moist heat causes cellular proteins (including enzymes) in the microorganisms to become
inactivated, and the cells die. Boiling water and steam are favored in disinfection, because no expensive equipment
is necessary and the required time is short. Most pathogens die after 10 minutes of steaming at 700C; also, boiling
for 10 to 30 minutes at 900 to 1000C, depending on the altitude, destroys most viable bacteria, fiangi, and viruses.
Clean articles made of metal and glass, such as syringes, needles, and simple instruments, may be disinfected by
boiling for 30 minutes. However, ~s technique is not always effective, because heat-resistant bacterial endospores,
mycobacteria, and viruses may be present. As mentioned in Chapter 2, the endospores of the bacteria that cause
anthrax, tetanus, gas gangrene, and botulism, as well as hepatitis viruses, are notably heat resistant and often survive
normal disinfection procedures. Because the temperature at which water boils is lower at higher altitudes, water
should always be boiled for longer times at high altitudes.
An effective way to disinfect clothing, bedding, and dishes is to use hot water, above 600C,
with detergent or soap and to agitate the solution around the items. This combination of heat,
mechanical action, and chemical inhibition is deadly to most pathogens.
Pressurized Steam An autoclave is a large metal pressure cooker that uses steam under pressure to completely
destroy all microbial life. Pressure raises the temperature of the steam and shortens the time necessary to sterilize
materials that can tolerate the high temperature and moisture. Autoclaving at a pressure of 15 p.s.i. at a tempera ture
ofl2l .50C for 20 minutes kills viable microorganisms, viruses, and exposed bacterial endospores, if they are not
protected by pus, feces, vomitus, blood, or other proteinaceous substances. Some types of equipment and certain
materials, such as
Moist Heat. As previously stated, moist heat causes cellular proteins (including enzymes) in the microorganisms to
become inactivated, and the cells die. Boiling water and steam are favored in disinfection, because no expensive
equipment is necessary and the required time is short. Most pathogens die after 10 minutes of steaming at 700C;
also, boiling for 10 to 30 minutes at 900 to 1000C, depending on the altitude, destroys most viable bacteria, fiangi,
and viruses. Clean articles made of metal and glass, such as syringes, needles, and simple instruments, may be
disinfected by boiling for 30 minutes. However, this technique is not always effective, because heat-resistant
bacterial endospores, mycobacteria, and viruses may be present. As mentioned in Chapter 2, the endospores of the
bacteria that cause anthrax, tetanus, gas gangrene, and botulism, as well as hepatitis viruses, are notably heat

resistant and often survive normal disinfection procedures. Because the temperature at which water boils is lower at
higher altitudes, water should always be boiled for longer times at high altitudes.
An effective way to disinfect clothing, bedding, and dishes is to use hot water, above 600C,
with detergent or soap and to agitate the solution around the items. This combination of heat,
mechanical action, and chemical inhibition is deadly to most pathogens.
Pressurized Steam. An autoclave is a large metal pressure cooker that uses steam under pressure to completely
destroy all microbial life. Pressure raises the temperature of the steam and shortens the time necessary to sterilize
materials that can tolerate the high temperature and moisture. Autoclaving at a pressure of 15 p.s.i. at a tempera ture
of 121.50C for 20 minutes kills viable microorganisms, viruses, and exposed bacterial endospores, if they are not
protected by pus, feces, vornitus, blood, or other proteinaceous substances. Some types of equipment and certain
materials, such as
rubber, which may be damaged by high temperatures, can be autoclaved at lower temperatures for longer periods.
The timing must be careftilly determined based on the contents and compactness of the load. All articles must be
properly packaged and arranged within the autoclave to allow steam to penetrate each package. Cans should be
open, bottles covered looselv with foil or cotton, and instruments wrapped in cloth. Sealed containers should not be
autoclaved. Pressure-sensitive autoclave tape (Fig. 7-4) and commercially available solutions containing bacterial
spores can be used as quality control measures to ensure that autoclaves are flinctioning properly.
COLD
Most microorganisms are not killed by cold temperatures and freezing, but their metabolic activities are slowed,
greatly inhibiting their growth. Thus, freezing is a microbistatic method of preservation in which the
microorganisms are in a state similar to suspended animation. when the temperature is raised above the freezing
point, the metabolic reactions speed up and the organisms slowly begin to reproduce again. Refrigeration merely
slows the growth of microorganisms; it does not altogether inhibit them. Many foods, biologic specimens, and
bacterial cultures are preserved by rapid freezing to very low temperatures, using liquid nitrogen. It should be noted
that slow freezing causes ice crvstals to form within cells and may rupture the cell membranes and cell walls of
some bacteria; hence, if it is important to preserve a pure culture of bacteria, such slow freezing should be avoided.
Persons who are involved in the preparation and preservation ~ foods must be aware that
thawing to room temperature allows bacterial spores to germinate and microorganisms to resume
growth. Consequently, refreezing of thawed foods is an unsafe practice, because it preserves the
millions of microbes that might be present and the food deteriorates quickly when it is rethawed.
Also, if bacterial endospores
of C. hotuhunin or C. penfringens were present, the viable bacteria would begin to produce toxins that would cause
food poisoning.
DRYING
For manv centuries, foods have been preserved by dr~~ng. Whcn moi~ture and nutrients are lacking, many dried
microorganisms remain viable, although they cai~
not
, , antitoxins, antibiotics, and pure cultures of
reproduce. Foods, antisera toxins
microorganisms are often preser\~d by lyophilization (discussed previously).

are professionals should keep in mind that dried viable pathogens may be lurking in dried matter, including blood, pus, fecal material,
and dust that are found on floors, in bedding, on clothing, and in wound dressings. Should these dried materials be
disturbed, such as by dry dustbv con
mg, the microbes would be easily transmitted through the air or
tact. They
would then grow rapidly if thev settled in a suitably moist, warm, nutrient environment such as a wound or a burn.
Therefore, important precautions must be obse~~d, including the following: wet mop and damp dust floors and
flirniture, roll the bed linens and towels carefially, and properly dispose of wound dressings.
RADIATION
The sun is not a particularly reliable disinfecting agent because it kills onlv those microorganisms that are exposed to
direct sunlight. The ravs of the sun include the long infrared (heat) rays, the visible light rays, and the shorter
ultraviolet (UV) rays. The UV rays, which do not penetrate glass and building materials, are effective only in the air
and on the surface of equipment. They do, however, penetrate cells and, thus, can cause damage to DNA. when this

occurs, genes may be so severely damaged that the cell dies (especially unicellular microorganisms) or is drastically
changed.
In practice, a UV lamp is useful for reducing the number of microorganisms in the air. A UV
lamp is often called a germicidal lamp. Its main component is a low-pressure mercury vapor tube.
Such lamps are found in newborn nurseries, operating rooms, elevators, entry ways, cafeterias,
and classrooms, where they are incorporated into louvered ceiling fixtures designed to radiate
across the top of the room without striking persons in the room. The sterility of an area may also
be maintained by having a UV lamp placed in a hood or cabinet containing instruments, paper and
cloth equipment, liquid, and other inanimate articles. Many biologic materials, such as sera,
antisera, toxins, and vaccines, are sterilized with UV rays.
Those whose work invok'es the use of UV lamps must be particularlv careful not to expose
their eves or skin to the rays, because they can cause serious burns and cellular damage. Because
UV rays do not penetrate cloth, metals, and glass, these materials may be used to protect persons
working in a UV environment. It has been shown that skin cancer can be caused bv excessive
exposure to the UV rays of the sun; thus, extensive suntanning is harmful.
X-rays and gamma and beta rays of certain wavelengths from radioactive materials may be
lethal or cause mutations in microorganisms and tissue cells, becatise
thcv damage DNA and proteins within those cells. Studies performed in radiation research
laboratories have demonstrated that these radiations can be used for the prevention of food spoilage,
sterilization of heat-sensitive surgical e4~ipment, preparation of vaccincs, and treatment of some
chronic discases such as C~&Ct~ all of which are very practical applications for lab()rator~ research.
The Food and Drug Administration approved the usc of gamma rays (from cobalt-60) to process
chickens and red meat in 1992 and 1997, respectively. Since then, gamma rays have been used by
some food processing plants to kill pathogcns (like Salmonella and Campyh)bactcr spp.) in chickens,
which are labeled "irradiated" and bear the green inter-national symbol for radiation. When radiations
are used in the treatment of disease, care must be taken to focus the rays precisely on the specific area
being treated to minimize damage to surrounding normal cells.
ULTRASONIC WAVES
In hospitals and clinics, ultrasonic waves are a frequentlv used means of cleaning and sterilizing
delicate equipment. Ultrasonic cleaners consist of tanks filled with liquid solvent (usually water); the
short sound waves are then passed through the liquid. The sound waves mechanic~v dislodge organic
debris on instruments and glassware.
Glassware and other articles that have been cleansed in ultrasonic equipment must be then
washed to remove the dislodged particles and solvent, and then sterilized by another method before
they are used.
FILTRATION
Filters of various pore sizes are used to filter or separate cells, larger viruses, bacteria, and certain
other microorganisms from the liquids or gases in which they are suspended. The filtered solution
(filtrate) is not necessarily sterile, because small viruses mav not be filtered out. The variety of filters
is large and includes sintered glass (in which uniform particles of glass are fused), plastic films,
unglazed porcelain, asbestos, diatomaceous earth, and cellulose membrane filters. Small quantities of
liquid can be filtered through a syringe; large quantities require larger apparatuses.
A cotton plug in a test tube, flask, or pipette is a good filter for preventing the entrv of
microorganisms. Dry gauze and paper masks prevent the out\~~ard passage of microbes from the
mouth and nose, at the same time protecting the wearer from inhaling airborne pathogens and foreign
particles that could damage the lungs. Biological safety cabinets and laminar flow hoods contain high
efficiency particulate air (HFPA) filters to protect workers from contamination.

Chemical Antimicrobial Methods

Chemical disinfection means the use of chemical agents to inhibit the growth of microorganisms,
either temporarily or permanently. The effectiveness of a chemical disinfectant depends on many
factors: the concentration of the chemical; the

time allowed for the chemical to work (called contact time); the pH of the solu tion; the temperature; and the
presence ofprote ins, blood, pus, feces, mucous secretions, and vomitus. Directions for the preparation and dilution
of the disinfectant must be carefully ft)llowed, and the proper concentration, pH, and temperature must be
maintained for the specified time period to ensure the best results. The items to be disinfected must first be washed
to remove any proteinaceous material in which pathogens may be hidden. Although the washed article may then be
clean, it is not safe to use until it has been properly disinfected. Healthcare personnel need to understand an
important limitation of chemical disinfection: many disinfectants that are effective against pathogens in the
controlled conditions of the laboratory become ineffective in the actual hospital or clinical environment.
Furthermore, the stronger and more effective antimicrobial chemical agents are of limited usefulness because of
their destructiveness to human tissues and certain other substances.
Almost all bacteria in the vegetative state as well as fungi, protozoa, and most viruses are
susceptible to many disinfect ants, although the mycobacteria that cause tuberculosis and leprosy,
bacterial endospores, pseudomonads, fungal spores, and hepatitis viruses are notably resistant.
Therefore, chemical disinfection should never be attempted when it is possible to use proper
physical sterilization techniques.
The disinfectant most effecti~~ for each situation must be carefully chosen. Chemical agents
used to disinfect respiratory therapy equipment and thermometers must destroy all pathogenic
bacteria, fungi, and viruses that may be found in sputum and saliva. One must be particularly
aware of the oral and respiratory pathogens, including Mycobacterium tuberculosis species of
Pseudomonas, Staphylococcus, and Streptococcus, the various fungi that cause candidiasis,
blastomycosis, cocci dioidomycosis, and histoplasmosis, and all of the respiratory viruses.
Because most disinfection methods do not destroy all bacterial endospores that are present,
any instrument or dressing used in the treatment of an infected wound or a disease caused by
spore-formers must be autoclaved or incinerated. Gas gangrene, tetanus, and anthrax are examples
of diseases caused by spore-formers that require the health worker to take such precautions.
Formaldehyde and ethylene oxide, when properly used, are highly destructive to spores,
mycobacteria, and viruses. Certain articles are heat sensitive and cannot be autoclaved or safely
washed before disinfection; such articles are soaked for 24 hours in a strong detergent and disin fectant solution, washed, and then sterilized in an ethylene oxide autoclave. The use of disposable
equipment whenever possible in these situations helps to protect patients and healthcare team
members.
The effectiveness of a chemical agent depends to some extent on the physical characteristics
of the article on which it is used. A smooth, hard surface is readily disinfected, whereas a rough,
porous, or grooved surface is not. Thought must be given to selection of the most suitable
germicide for cleaning patient rooms and all other areas where patients are treated.
The most effective antiseptic or disinfectant should be chosen for the specific
purpose, environment, and pathogen or pathogens likely to be present. The characteristics of a good chemical
antimicrobial agent are as follows:
It must kill pathogens within a reasonable period and in specified concentrations.
It must be nontoxic to human tissues and noncorrosive and nondestructive to materials on which it is
used.
It must be soluble in water and easy to apply. If a tincture (e.g., alcohol-water solution) is used, the proper
concentration must be used. Evaporation of the alcohol solvent can cause a 1% solution to increase to a 10%
solution, and at this concentration, it may cause tissue damage.
It should be inexpensive and easy to prepare for use with simple, specific directions.
It must be stable in the dissolved or solid form so that it can be shipped and stored for a reasonable
period.
It should be stable to pH and temperature changes within reasonable limits.
ANTISFPSIS
Most antimicrobial chemical agents are too irritating and destructive to be applied to mucous membranes and skin.
Those that may be safely used on human tissues are called antiseptics. An antiseptic merely reduces the number of
organisms on a surface but does not penetrate the pores and hair follicles to destroy microorganisms residing there.
To remove organisms lodged in pores and folds of the skin, health personnel use an antiseptic soap and scrub with a

brush. To prevent resident indigenous microfiora from contaminating the surgical field, surgeons wear sterile gloves
on freshly scrubbed hands and masks and hoods to cover face and hair. Also, an antiseptic is applied at the site of the
surgical incision to destroy local microorganisms.
HOW ANTIMICROBIAL CHFMICALS WORK
Injury of Cell Membranes. Soap and detergents are referred to as surfactants; this means that they are surface-active
agents that help to disperse bacteria, allowing them to more readily be rinsed away. These agents concentrate on the
surface and, thus, reduce the surface tension; this characteristic makes them good wetting and dispersing agents.
Some agents, such as Dial and 5afeguard soaps, contain disinfectants, which also aid in killing bacteria. Certain
concentrations ofweak acids such as acetic and benzoic acids may also be used in disinfectant soaps.
Inactivation ofEnzymes. Alcohols, such as ethyl and isopropyl, are good skin antiseptics at 70% solution. Ethyl
alcohol has a low toxicity for humans; hence, it is frequently used to disinfect clinical thermometers and other
instruments. However, when taken internally, isopropyl alcohol causes severe gastrointestinal upset and methanol
causes brain damage. Alcohols are tuberculocidal (destructive to tuberculosis-causing organisms), but not sporicidal
(destructive to spores).
The phenolics including phenol, carbolic acid, xylenols, orthophenylphenol, and cresol are used as disinfectants in
hospitals and laboratories. However, they are too irritating and toxic to be used on skin. The commercial mixture of
phenolics, Lysol, is an effective germicide because it works in the presence of organic material and remains acuve
on hard surfaces for extended periods. These chemicals are tuberculocidal, but not sporicidal.
The effectiveness of phenol was demonstrated by Joseph Lister in 1867, when it was used to reduce the incidence
of infections ft)llowing surgical procedures. The effectiveness of other disinfectants is compared with that of
phenol using tile phenol coefficient test. To perform this test, a series of dilutions of phenol and the experimental
disinfectant are inoculated with the test bacteria, Salmonella typhi and Staphylococcus aureus, at 370 ~ The
highest dilutions (lowest concentrations) that lull the bacteria after 10 minutes are used to calculate the phenol
coeflicient.
Salts of hea~~ metMs such as mercury chloride (Meithiolate, Mercuroebrome, Metaphen~genen.c names:
thimerosal, merbromin, nitrom ersol, respectively) and silver nitrate (Argyrol, protargol) are bacteriostatic
antiseptics, but thev are not sporicidal and are ineffective against many pathogens. In the past, silver nitrate in
low concentrations was used in the eyes of newborns to lull Neisseriagonorrhoeae, thus prevenring gonococcal
eve infections which could cause blindness.
Chemical oxidizing agents are usefal disinfectants. Two of these are hydrogen peroxide and sodium perborate,
which destroy bacteria and tissue debris and pre~~nt growth of anaerobes in damaged tissues. A third, potassium
permanganate, is used in weak solutions to treat urethral infections and fungal infections of the skin. Another
agent in this group is ethylene oxide (Carboxide, Cr,Toxide, Oxygume), which is used as a sterilant in gas
autoclaves to sterilize heat-sensitive materials. Although it is a good microbicide and sporicide, this gas must be
used with great care because it is flammable and toxic to humans.
The many Compounds of chlorine, iodine, bromine, and fluorine are also useful disinfectants. For example,
chlorine compounds (CIorox, Halozone, hypochlorites, Warexin) are used to disinfect water and sewage
and f~)r sanitization of dishes, floors, and plumbing fixtures. It has been found that human immunodeficiency
virus (HIV) can be destroyed on syringes and needles by soakmg them in a 1:10 Clorox (chlorine bleach)
solution for 10 minutes. Iodine compounds, such as Wescodyne, Betadine, Isodine, and tincture of iodine
are effective skin antiseptics and disinfectants. However, these compounds can be dangerous. If an alcohol
solution of iodine is left open to the air, allowing the alcohol to evaporate, the solution mav become too
concentrated, and an iodine burn may result if it is used on skin. Most compounds of bromine and fluorine are
too toxic at the effective concentrations to be used as antiseptics. All of these compounds are viricidal,
bactericidal, and tuberculocidal; however, none is sporicidal.
Damage to Genetic Material. The DNA of cells is inactivated by caustic compounds such as formalin. Formalin is a
37% aqueous solution of gaseous formaldehyde that in-activates proteins and nucleic acids. It is one of the few
antimicrobial agents that are also sporicidal; however, it is so irritating to skin and muc~~us membranes that it
CHEMOTHERAPY

Chemotherapeutic agents are chemical substances (drugs) used to treat diseases, including infectious
diseases. For thousands of years, people have been finding and using herbs and chemicals to cure
diseases. Native witch doctors in Central and South America long ago discovered that the herb,
ipecac, aided in the treatment of dysentery and that a quinine extract of cinchona bark was effective
in treating malaria. During the 16th and 17th centuries, the alchemists of Europe searched for wavs to
cure smallpox, syphilis, and many other diseases that were rampant during that period of history.
Many of the mercury and arsenic chemicals that were used frequently caused more damage to the
patient than to the pathogen. Chemotherapeutic agents used to treat infectious diseases are called
antimicrobial agents.
Characteristics of Antimicrobial Agents
An antimicrobial agent is any chemical (drug) used to treat an infectious disease by inhibiting or
killing pathogens in vivo (in the living animal). Some antimicrobial agents are antibiotics. An
antibiotic is a substance produced by a microorganism that is effective in killing other
microorganisms. Antibiotics are produced by certain molds and bacteria, usually those that live in
soil. Penicillin is an example of an antibiotic produced by a mold and bacitracin is an example of an
antibiotic produced by a bacterium. Although originally produced by microorganisms, many
antibiotics are now synthesized or manufactured in pharmaceutical laboratories. Also, many
antibiotics have been chemically modified to kill a wider variety of pathogens; these modified
antibiotics are called semisynthetic antibiotics.
The ideal antimicrobial agent should (1) kill or inhibit the growth of pathogens, (2) cause no
damage to the host, (3) cause no allergic reaction in the host, (4) be stable when stored in solid or
liquid form, (5) remain in specific tissues in the body long enough to be effective, and (6) kill the
pathogens before they mutate and become resistant to it. However, most antimicrobial agents have
some side effects, produce allergic reactions, or permit development of resistant mutant pathogens.
How Antimicrobial Agents Work
To be acceptable, an antimicrobial agent must inhibit or destroy the pathogen without damaging the
host. To accomplish this, the agent must target a metabolic process or structure possessed by the
pathogen, but not possessed by the host. The following examples illustrate this principle.
Sulfonamide drugs inhibit production of folic acid in those bacteria that require paraaminobenzoic acid (PABA) to synthesize folic acid. Folic acid (a vitamin) is Cssential to these
bacteria. Because the sulfonamide molecule is similar in shape to the PABA molecule, bacteria
attempt to metabolize sulfonamide to produce folic acid (Fig. 7-6). However, the enzymes that convert
PABA to folic acid cannot produce folic acid from the sulfonamide molecule. Without folic acid,
bacteria cannot produce certain essential proteins and finally die. Sulfa drugs, therefore, are called
competitive inhibitors; that is, by competing with an enzyme that metabolizes an essential nuttient,
they inhibit growth of microorganisms. They are, therefore, bacteriostatic. Cells of humans and
animals do not synthesize folic acid from PABA; they get folic acid from the food they eat.
Consequently, they are unaffected bv sulfa drugs.
In most Gram-positive bacteria, including streptococci and staphylococci, penicillin interferes
with the synthesis of the peptidoglycan that is required in bacterial
cell walls. Thus, by inhibiting cell wall synthesis, penicillin destroys the bacteria.
~y doesn't penicillin also destroy human cells? Because human cells do not have
a cell wall.
There are other antimicrobial agents that have similar action; they inhibit a specific step that is
essential to the microorganism's metabolism and thereby cause its destruction. Antibiotics are used in
this way against bacteria and they are highly effective. Some specifically destroy Gram-positive
bacteria; others specifically destroy

Gram-negative bacteria. Those that are destructive to both Gram-positive and Gram-negative bacteria are called
broad-spectrum antibiotics. Examples of broad-spectrum antibiotics are ampicillin, chloramphenicol, and
tetracvcline. Table 7-2 lists some of the antimicrobial drugs most frequently used against many common
pathogens.
Antimicrobial agents work well against bacteri~ pathogens because the bacteria (being procarvotic) have different
cellular structures and metabolic pathways that can be disrupted or destroyed by drugs that do not damage the host's
(eucaryotic) cells. Bactericidal agents kill bacteria, whereas bacteriostatic agents stop them from growmg and
dividing. Bacteriostatic agents should olily be used in patients whose host defense mechanisms (Chapter 11) are
frinctioning properly. Some of the mechanisms by which antibacterial agents kill or inhibit bacteria are shown in
Table 7-3.
Frequenily, a single antimicrobial agent is not sufficient to destrov all the pathogens that develop during the course
of a disease; thus, ~vo or more drugs ma~ be used simultaneously to kill all of the pathogens and to prevent resistant
mutant pathogens from emerging. In tuberculosis, for example, four drugs (isoniazid, rif ampin, pyrazinamide, and
either ethambutol or streptomycin) are routinely prescribed, and as many as 12 drugs may be required for especiallv
resistant strains. Many urinary, respiratory, and gastrointestinal infections respond particularlv well to a combination
of trimethoprim and sulfamethoxazole, a combination referred to as co-trimoxazole.
It is much more difficult to use antimicrobial drugs against fungal and protozoal pathogens because they are
eucaryotic cells; thus, the drugs are much more toxic to the host. Most antiflingal agents work in one of three ways:
(1) by binding with cell membrane sterols; e.g., nystatin and amphotericin B; (2) by interfering with sterol synthesis;
e.g., clotrimazole and miconazole; or (3) by blocking mitosis or nucleic acid synthesis; eg., griseoflilvin and 5flucytosine. Antiprotozoal drugs are usually quite toxic to the host and work by (1) interfering with DNA and RNA
synthesis; eg., chloroquine, pentamidine, and quinacrine; or (2) interfering with protozoal metabolism; e.g.,
metronidazole (Flagyl).
Antiviral chemotherapeutic agents are particularly difficult to develop and use because viruses are produced within
host cells. A frw drugs have been found to be effective in certain viral infections; these work by inhibiting viral
replication within cells. Some antiviral chemotherapeutic agents are listed in Table 7-4.
In cancer, in which malignant cells are dividing more rapidly than normal cells, chemical agents that inhibit DNA
and BNA synthesis can be used, provided the dosage and total period of administration are careni fly controlled.
Cancer drugs interfere with normal DNA fanction in rapidly dividing cells, regardless of whether the cells are
normal or malignant. Thus, normal cells that are rapidly dividing, including skin cells, erythroblasts that later
become red blood cells, and sperm cells, are damaged along with the malignant cells. This is why blood cell counts
are performed frequenthi in cancer patients; the physician must be able to determine at