Lecture : Introduction to Bacteria part3

Instructor: Roy P. Maribojoc MD, MPH

Bacterial Reproduction

1. Binary Fission: bacterial cell divides to form two identical cells

o A form of Asexual Reproduction since it involves only one parent

o offspring are identical to the parent

2. Conjugation

o A form of sexual reproduction since it involves two bacterial parents

o genetic information is exchanged between organisms
o does not increase the number of bacteria
o results in bacteria that are genetically different

1
Spore formation
o Spore: a small, rounded, thick-walled, resting cell that forms inside a bacterial
cell when conditions are not good enough to reproduce.
o Spore can respond quickly to changes in the environment returning to the
vegetative state immediately if condition becomes favorable
o Helps bacteria survive harsh conditions

Example : Clostridium tetani (tetanus)

Bacillus anthracis (anthrax)

Tetanus
is a serious illness caused by Clostridium tetani bacteria. Bacterial spores are found in
soil, which can remain infectious for more than 40 years. They also exist in
environments as diverse as animal saliva, manure, house dust, and the human colon. If
the spores enter the body through a deep cut, like those you might get from stepping on
a nail, or through a burn, they germinate and produce a toxin that enters the
bloodstream.
This toxin known as tetanospasmin is taken up from the blood by the outermost nerves
and moves inward toward the spine. After approximately eight days (ranging from three
to 21 days), it begins to short-circuit nerve signals and block the relaxation of muscles.
The neurotoxin disrupts the normal workings of the nerves, causing symptoms such as
stiffness and muscle spasms usually all over the body. It can lead to "locking" of the jaw
(the disease is also known as “lockjaw”). This makes it impossible to open your mouth
or swallow. Spasms of the jaw or facial muscles may follow, spreading to the hands,
arms, legs, and back and blocking the ability to breathe. Spasms are often precipitated
by noise or touch. Once tetanus has spread, the mortality rate is approximately 30%,
even in modern medical facilities. Tetanus is a medical emergency. You need to get
treatment in a hospital. A vaccine can prevent tetanus.
Neonatal tetanus is a form of generalized tetanus in newborn infants that do not have
protective passive immunity because the mother is not immune. It usually occurs

2
through infection of the unhealed umbilical stump, particularly when the stump is cut
with an unsterile instrument. Neonatal tetanus is estimated to kill over 200,000
newborns each year; almost all these deaths occur in developing countries
Anthrax is a disease caused by Bacillus anthracis, bacteria that live in soil. It affects
animals such as cattle, sheep, and goats more often than people. People can get anthrax
from contact with infected animals, wool, meat, or hides. It can cause three forms of
disease in people. They are

● Cutaneous, which affects the skin. People with cuts or open sores can get it if
they touch the bacteria.
● Inhalation, which affects the lungs. You can get this if you breathe in spores of
the bacteria.
● Gastrointestinal, which affects the digestive system. You can get it by eating
infected meat.

Symptoms of anthrax differ depending on the type of anthrax.

Symptoms of cutaneous anthrax start 1 to 7 days after exposure:

● An itchy sore develops that is similar to an insect bite. This sore may blister and
form a black ulcer (sore or eschar).
● The sore is usually painless, but it is often surrounded by swelling.

● A scab often forms, and then dries and falls off within 2 weeks. Complete healing
can take longer.

Symptoms of inhalation anthrax:

● Begins with fever, malaise, headache, cough, shortness of breath, and chest pain
● Fever and shock may occur later

Symptoms of gastrointestinal anthrax usually occur within 1 week and may include:

● Abdominal pain
● Bloody diarrhea
● Diarrhea
● Fever
● Mouth sores
● Nausea and vomiting (the vomit may contain blood)

3
Many people know about it from the 2001 bioterror attacks. In the attacks, someone
purposely spread anthrax through the U.S. mail. This killed five people and made 22 sick.

Gram Staining
o Developed by Hans Christian Gram (a Danish doctor) in 1884
use to differentiate two large groups of bacteria based on the chemical and physical
properties of their cell walls: Gram (+) and Gram ( -) bacteria
o Procedure:
Step 1 Crystal Violet 60 seconds
Step 2 Water Rinse
Step 3 Gram’s Iodine 60 seconds
Step 4 Water Rinse
Step 5 95% Ethanol 10 seconds
Step 6 Water Rinse
Step 7 Safranin (counter stain) 60 seconds
Step 8 Water Rinse
Step 9 Dry The Slide

o Result:
o Gram (+) bacteria= PURPLE
o Gram ( -) bacteria= PINK

Gram reaction is based on the structure differences of the bacterial cell wall:

4
 o In Gram-positive bacteria, the purple crystal violet stain is trapped in the THICK
layer of peptidoglycan which forms the outer layer of the cell; thus staining it
purple

o In Gram-negative bacteria, it has an additional outer membrane of

lipopolysaccharides that prevents the stain from totally penetrating the THIN
peptidoglycan layer. The outer membrane is then permeabilized or “destroyed”
by alcohol treatment, and allowing the pink safranin counterstain to be trapped
in the peptidoglycan layer; thus staining it PINK

Gram-negative bacteria commonly cause more serious infection than gram-positive

bacteria due to their lipopolysaccharide in their outer membrane which produces the
endotoxin that increases the severity of inflammation, and they are more resistant to
common antibiotic like penicillin because of its cell wall structure. This inflammation
may be so severe that septic shock may occur. Gram-positive infections are generally
less severe because humans can produce an enzyme (lysozyme) that attacks the open
peptidoglycan layer of Gram-positive bacteria. Gram-positive bacteria are also
frequently much more susceptible to beta-lactam antibiotics, such as penicillin.

5
6