Bacterial Spores- are highly resistant, dormant structures (i.e.

, no metabolic activity) formed in

response to adverse environmental conditions. They help in the survival of the organisms
during adverse environmental conditions; they do not have a role in reproduction.

Spore formation (sporulation) occurs when nutrients, such as sources of carbon and nitrogen

are depleted.  Bacterial spores are highly resistant to

 Heat
 Dehydration
 Radiation and
 Chemicals.

An endospore is structurally and chemically more complex

than the vegetative cell.  It contains more layers than
vegetative cells. Resistance of Bacterial spore may be
mediated by dipicolinic acid, a calcium ion chelator found
only in spores.  When the favorable condition prevails,
(i.e., availability of water, appropriate nutrients) spores
germination occurs which forms vegetative cells of
pathogenic bacteria.

Following factors/constituents plays major role for the

resistance of Bacterial Spore: 

 Calcium dipicolinate in core

 Keratin spore coat
 New enzymes (i.e., dipicolinic acid synthetase, heat-resistant catalase)
 Increases or decreases in other enzymes.

A mature endospore contains a complete set of the

genetic material (DNA) from the vegetative cell, ribosomes
and specialized enzymes.

The shape and the position of spores vary in different

species and can be useful for classification and
identification purposes. Endospores may be located in
the middle of the bacterium (central), at the end of the
bacterium (terminal) and near the end of the bacteria
(subterminal) and may be spherical or elliptical.
Spores may be:

1. Central or equatorial, giving the bacillus a spindle shape (eg. Clostridium

bifermentans)
2. Sub-terminal, the bacillus appearing Club shaped (eg. Clostridium perfringens)
3. Oval and terminal, resembling a tennis racket (eg. Clostridium tertium)
4. Spherical and terminal, giving a drumstick appearance (Clostridium tetani)

Mature endospores are released from the vegetative cell to become free endospores.  When
the free endospores are placed in an environment that supports growth, the endospores will
revert back to a vegetative cell in a process called germination.  It should be noted that unlike
the process of binary fission observed with vegetative cells, endospore formation is not a
reproductive process but a process of differentiation that provides the bacteria with a
mechanism for survival.

Constituents of Bacterial Spores:

1. Thick keratinlike coat

2. Peptidoglycan
3. Cell membrane
4. A small amount of cytoplasm
5. Very little water
6. Bacterial DNA

Medical Importance of Bacterial Spores

Important features of Spores Medical Implications

Spores are highly resistant to heating; spores are
not killed by boiling (100°C) but are killed at
121°C.
Spores are highly resistant to many chemicals,
including most disinfectants.
Spores can survive for many years in soil and
Medical supplies must be heated to 121°C for at
least 15 minutes to be sterilized.
Only solution designated as sporicidal will kill
spores.
Wound contaminated with soils can be infected
 with spores and cause diseases such as tetanus,
other inanimate objects. gas gangrene.
Spores do not exhibit measurable metabolic
activity. Antibiotics are ineffective against spores.
Spores formed only when nutrients are Spores are not often found at the site of
insufficient. infection because nutrients are not limiting.

Examples of Spore forming Bacteria- Spores formed by only two genera of Gram-positive rods
are of medical importance.

 Bacillus spp
 Clostridium spp

Sterilization refers to any process that removes, kills, or deactivates all forms of life (in
particular referring to microorganisms such
as fungi, bacteria, spores, unicellular eukaryotic organisms such as Plasmodium, etc.) and
other biological agents like prions present in a specific surface, object or fluid, for example food
or biological culture media. Sterilization can be achieved through various means,
including heat, chemicals, irradiation, high pressure, and filtration. Sterilization is distinct
from disinfection, sanitization, and pasteurization, in that those methods reduce rather than
eliminate all forms of life and biological agents present. After sterilization, an object is referred
to as being sterile or aseptic.
Disinfection:
Thermal or chemical destruction of pathogenic and other types of microorganisms. Disinfection
is less lethal than sterilization because it destroys most recognized pathogenic microorganisms
but not necessarily all microbial forms (e.g., bacterial spores).

Autoclave:
Device that sterilizes instruments or other objects using steam under pressure. The length of
time required for sterilization depends on temperature, vacuum, and pressure.

Sanitization can refer to Cleaning and Disinfection of an area or an item. The sanitizing is mostly
used in the food preparation areas and refers to eliminating or reducing bacteria by cleaning
and disinfecting.
fire extinguisher is an active fire protection device used to extinguish or control small fires,
often in emergency situations. It is not intended for use on an out-of-control fire, such as one
which has reached the ceiling, endangers the user (i.e., no escape route, smoke, explosion
hazard, etc.), or otherwise requires the expertise of a fire brigade. Typically, a fire extinguisher
consists of a hand-held cylindrical pressure vessel containing an agent that can be discharged to
extinguish a fire. Fire extinguishers manufactured with non-cylindrical pressure vessels also
exist but are less common.
There are two main types of fire extinguishers: stored-pressure and cartridge-operated. In
stored pressure units, the expellant is stored in the same chamber as the firefighting agent
itself. Depending on the agent used, different propellants are used. With dry chemical
extinguishers, nitrogen is typically used; water and foam extinguishers typically use air. Stored
pressure fire extinguishers are the most common type. Cartridge-operated extinguishers
contain the expellant gas in a separate cartridge that is punctured prior to discharge, exposing
the propellant to the extinguishing agent. This type is not as common, used primarily in areas
such as industrial facilities, where they receive higher-than-average use. They have the
advantage of simple and prompt recharge, allowing an operator to discharge the extinguisher,
recharge it, and return to the fire in a reasonable amount of time. Unlike stored pressure types,
these extinguishers use compressed carbon dioxide instead of nitrogen, although nitrogen
cartridges are used on low temperature (-60 rated) models. Cartridge operated extinguishers
are available in dry chemical and dry powder types in the U.S. and in water, wetting agent,
foam, dry chemical (classes ABC and B.C.), and dry powder (class D) types in the rest of the
world.

The Main Routes Infection can Enter the Body are:

Body fluids – A body fluid e.g. blood, urine, pus, saliva from one person enters the body of
another e.g. through cuts or other means such as sexual intercourse (e.g. syphilis, HIV etc.).
Through saliva (e.g. glandular fever). Through contaminated substances entering the
bloodstream e.g. tetanus can be contracted through wounds when handling contaminated soil.

Through the air – e.g. tuberculosis. Droplet infections are carried through the air through
coughs and sneezes (e.g. common cold, respiratory viruses).

Through touch – Those caught by directly touching the skin/skin to skin contact (these are
often referred to as contagious) of someone who has a particular kind of infection such as
chickenpox.

Touching an infected object which an infected person has touched e.g. door handles, paper and
cooking implements – e.g. Norovirus (winter vomiting infection) can be caught this way.

Through ingestion – contaminated food causes food poisoning (e.g. salmonella and
campylobacter) or water (e.g. cholera)
Through bites from other creatures – e.g. infections from dog and cat bites, (e.g. blood
poisoning/sepsis) or through parasites e.g. insect bites/stings (e.g. malaria). Insect bites and
stings can also cause allergic reactions which are not due to infections but due to toxins which
exist in the venom which has entered the body.

Communicable disease is an illness transmitted through contact with microorganisms .

Safety Station
emergency eyewash and safety shower station are essential equipment for every laboratory
that uses chemicals and hazardous substances. Emergency eyewash and safety shower stations
serve the purpose of reducing workplace injury and keeping workers away from various
dangers.[1]

Types
Safety Shower
A safety shower is a unit designed to wash an
individual's head and body which has come into contact with hazardous
chemicals. Large volumes of water are used and a user may need to take off
any clothing that has been contaminated with hazardous chemicals. Safety
showers cannot be used for flushing an individual's eyes, due to the high
pressure of water from the shower, which can damage a user's eyes.

Eyewash Station
An eyewash station is a unit for washing chemicals or substances
that might splash into an individual's eyes before he or she can seek
further medical attention. The user needs to wash their eyes for at
least 15 minutes.[2]

Drench Hose
A drench hose is an equipment that can spray water to a specific spot of the
chemical exposure on individual's body. The benefit of a drench hose is that it can
be applied to an individual who cannot reach a normal eyewash or shower station or
in the case where the eyewash and shower station are unavailable.
Combination Unit
A combination unit is where other units such as a shower station, eyewash station, and drench
hose share the same water supply plumbing. This unit is useful in laboratory where hazardous
chemicals with different properties are used.
Eyewash Bottle
Also known as a personal eyewash unit, it is a supplementary for eyewash stations. However,
eyewash stations cannot be replaced by eyewash bottles since they do not meet safety
standards. Eyewash bottles allow an individual to flush the injured area immediately, or until
the individual can reach the fixed eyewash station. Early eyewashes were designed with a single
rinsing stream, but recent advancements have made eyewashes capable of flushing both eyes
simultaneously. A pH neutral solution for emergency eyewash[3] may also be chosen to reduce
the danger from contaminants if strong acids or alkali chemicals are presented.
Safety Shower
The path from the hazard to the safety shower shall be free of obstructions
and tripping hazards.

 Water supply should be enough to provide at least 20 gallons per

minute of water for 15 minutes (Section 4.1.2, 4.5.5).
 Hand free valve should be able to open within one second and
remain open until it is manually closed (Section 4.2, 4.1.5).
 The top of the water column shall not be lower than 82"
(208.3 cm) and no higher than 96" (243.8 cm) above the surface
floor the user is standing on(Section 5.1.3, 4.5.4).
 Center of the water column should be at least 16" (40.6 cm) away from any
obstruction (Section 4.1.4, 4.5.4).
 Actuator should be easily accessible and easily located. It should be no more than
69" (173.3 cm) above the surface floor the user is standing on (Section 4.2).
 At 60" (152.4 cm) above the floor, the water pattern should be 20" (50.8 cm) in
diameter (Section 4.1.4).
 If shower enclosure is provided. It should provide 34" in diameter of unobstructed
space (86.4 cm) (Section 4.3).
 Water temperature of safety shower station should be within 60 °F - 100 °F (16 °C -
38 °C).
 Safety shower stations should have highly visible and well lit signage.
Eyewash Station
The path from the hazard to the Eyewash or Eye/Face wash shall be free of obstructions and
tripping hazards.

 Eyewash station shall flush both eyes simultaneously within gauge guidelines
(Eyewash gauge detailed in ANSI/ISEA Z358.1-2014) (Section 5.1.8).
  Eye or Eye/Face wash shall provide a controlled flow of water that is non-injurious
to the user (Section 5.1.1).
 Nozzles and flushing fluid shall be protected from airborne contaminants (dust
covers), and shall not require a separate motion by the operator when activating the
equipment (section 5.1.3).
 Eyewashes must deliver 0.4 gpm for 15 minutes, Eye/Face washes must provide 3
gpm for 15 minutes.
 The top of the Eye or Eye/Face wash water flow must not fall below 33" (83.8 cm)
and can be no higher than 53" (134.6 cm) from the floor surface floor the user is
standing on (Section 5.4.4).
 The head or heads of the Eyewash or Eye/Face wash must be 6" (15.3 cm) away
from any obstructions (Section 5.4.4).
 The valve must allow for 1 second operation and the valve shall remain open
without the use of the operator's hands until intentionally closed. (Section 5.1.4,
5.2).
 Manual or automatic actuators shall be easy to locate and readily accessible to the
user (Section 5.2).
 Water temperature of Eye or Eye/Face wash station should be within 60 °F - 100 °F
(16 °C - 38 °C).
 Eye or Eye/Face wash stations should have highly visible and well lit signage.

Location

Safety showers and eyewash stations should be within 10 seconds walking distance or 55 feet
(appendix B) from the hazard and must be located on the same level as the hazard, so the
individual does not have to go up or down the stairs when an accident occurs. Moreover, the
path way should be clear and free of obstructions.
Venipuncture or venepuncture is the process of obtaining
intravenous access for the purpose of venous blood sampling (also
called phlebotomy) or intravenous therapy. In healthcare, this
procedure is performed by medical laboratory scientists, medical
practitioners,
some EMTs, paramedics, phlebotomists, dialysis technicians, and
other nursing staff.[1] In veterinary medicine, the procedure is
performed by veterinarians and veterinary technicians.
It is essential to follow a standard procedure for the collection of
blood specimens to get accurate laboratory results. Any error in
collecting the blood or filling the test tubes may lead to erroneous
laboratory results.[2]
Venipuncture is one of the most routinely performed invasive procedures and is carried out for
any of five reasons:
 1. to obtain blood for diagnostic purposes;
2. to monitor levels of blood components;[3]
3. to administer therapeutic treatments including medications, nutrition, or
chemotherapy;
4. to remove blood due to excess levels of iron or erythrocytes (red blood cells); or
5. to collect blood for later uses, mainly transfusion either in the donor or in
another person.
Vacutainer blood collection tube is a sterile glass or plastic test tube with
a colored rubber stopper creating a vacuum seal inside of the tube,
facilitating the drawing of a predetermined volume of liquid. Vacutainer
tubes may contain additives designed to stabilize and preserve the
specimen prior to analytical testing. Tubes are available with a safety-
engineered stopper, with a variety of labeling options and draw volumes.
The color of the top indicates the additives in the vial.
Vacutainer tubes were invented by Joseph Kleiner and Becton
Dickinson in 1949. Vacutainer is a registered trademark of Becton
Dickinson, which manufactures and sells the tubes today.

TYPES
OF ANTICOAGULANTS AND PURPOSE

Vacutainer/sample tube types for venipuncture/phlebotomy   

Tube cap
color or Additive Usage and comments
type

Usually drawn first for minimal risk

of contamination.[8] Two bottles are
Blood Sodium polyanethol
typically collected in one blood
culture bott sulfonate (anticoagulant) and growth
draw; one for aerobic
le media for microorganisms
organisms and one for anaerobic
organisms.[9]
 Coagulation tests such
as prothrombin time (PT)
Light blue Sodium citrate (anticoagulant) and partial thromboplastin
time (PTT) and thrombin time (TT).
Tube must be filled 100%.

Serum: Total complement
Plain red No additive
activity, cryoglobulins

Serum-separating tube: Tube

inversions promote clotting. Most
Gold Clot activator and serum separating gel[10]
chemistry, endocrine and serology
tests, including hepatitis and HIV.

Chromosome
Dark green Sodium heparin (anticoagulant) testing, HLA typing, ammonia, lactat
e

Plasma. Tube inversions prevent

Mint green Lithium heparin (anticoagulant)
clotting

Whole blood: CBC, ESR, Coombs
Lavender test, platelet antibodies, flow
EDTA (chelator / anticoagulant)
("purple") cytometry, blood levels
of tacrolimus and cyclosporin

Blood typing and cross-matching,
Pink EDTA (chelator / anticoagulant)
direct Coombs test, HIV viral load

Trace elements, heavy metals, most

Royal blue EDTA (chelator / anticoagulant)
drug levels, toxicology

Tan EDTA (chelator / anticoagulant) Lead

  Sodium fluoride (glycolysis inh
ibitor)
Gray Glucose, lactate[12]
 Potassium oxalate (anticoagul
ant)[11]
Tissue typing, DNA
Yellow Acid-citrate-dextrose A (anticoagulant)
studies, HIV cultures
Pearl PCR for adenovirus, toxoplasma and 
Separating gel and (K2)EDTA
("white") HHV-6

SITE OF COMON VEIN PUNCTURE

Median Cubital Vein
The most commonly used site for a venipuncture is at the bend
of the elbow, also called the antecubital fossa. Several veins in
the antecubital fossa provide excellent access. The median
cubital vein, a superficial vein that forms a connection point
between the cephalic and basilic veins of the arm is large, and
therefore easier to see and feel. Risks associated with using this
vein include penetration of the biceps tendon and the brachial
artery. These tissues lie just below the median cubital vein so
care must be taken not to puncture too deeply.

Cephalic Vein
The cephalic vein runs superficially under the skin along the
outer side of the upper and lower arm, eventually dipping
under the shoulder to join the axillary vein in the armpit. This is
another acceptable venipuncture site if scarring is present in
the antecubital space from previous venipunctures or if the
antecubital veins are not palpable. The cephalic vein is more commonly accessed on the
forearm rather than the upper arm. You can also access the cephalic sign in the inside of the
wrist as it runs down to the thumb.

Basilic Vein
The basilic vein, another main superficial vein of the arm, runs along the inner side of the upper
arm and forearm. The basilic vein is also commonly used for a venipuncture in the forearm
rather than the upper arm. This is because halfway up the upper arm, the vein turns inwards to
become a deep vein. Deep veins run along arteries and present a risk for puncture to the artery.

Dorsal Metacarpal Veins

The veins that run down the middle of the hand, called dorsal
metacarpal veins, can also be used for intravenous infusions
as well as for drawing blood if the antecubital vein isn't
accessible. Hand veins aren't as large as the veins in the
forearm and antecubital fossa. In some hospitals, it's policy to
start with the veins on the hand as a first choice for an
intravenous line and work your way up if you can't access a
lower vein.

Hemolysis: The destruction of red blood cells which leads to the release of hemoglobin from

within the red blood cells into the blood plasma.

Etymology: The word "hemolysis" is made up of "hemo-", blood + "lysis", the disintegration of

cells.

Icterus (or the icterus index) is a measure of the yellow colour of serum. This colour is normally
due almost exclusively to the presence of bilirubin, a hemoglobin waste product from the red
blood cells. The icterus index is expressed as a number of “plus” signs (from zero to ++++). An
icterus index of zero is normal. A non-negative result (+ to ++++) indicates a concentration of
bilirubin above the normal value. To better understand what these results mean, see your
bilirubin result. Very high results (++++) can interfere with certain laboratory tests. More rarely,
a high icterus index can be caused by excessive consumption of foods like carrots or vitamin
supplements containing beta carotene.
Lipemia is a measure of serum transparency. High levels of blood lipids, mostly triglycerides,
increase serum turbidity. The lipemia result is expressed in “number of plus signs,” ranging
from negative to four pluses (++++). Too high of a concentration of triglycerides in the serum (+
+++) can interfere with some laboratory tests, particularly the measurement of sodium levels.
Some individuals have particularly high triglyceride levels after meals, and in some cases the
triglycerides will remain present for more than 24 hours. To better understand what these
results mean, see your triglyceride result.
Complications that can arise from venipuncture include:

 haematoma formation
 nerve damage, pain
 haemaconcentration
 extravasation
 iatrogenic anaemia
 arterial puncture
 petechiae
 allergies
 fear and phobia
 Infection
 syncope and fainting
 excessive bleeding
 edema and thrombus.