GROWTH AND NUTRITION OF MICROBES

Main constituent of bacteria- water (80% of total weight). Rest of them are made of
proteins, lipids, polysaccharides, nucleic acids, mucopeptides etc.

 Minimum nutrition for growth is water for growth and multiplication of

bacteria.
Water is the vehicle for the entry of all nutrients in to cells and for elimination
of all waste products.

Nutritional classification of bacteria based on their energy requirements and on

their ability to synthesis essential metabolites.

1. Phototrophs
Derive energy from sunlight.
2. Chemotrophs
Obtain energy from chemical reactions.
3. Autotrophs
 Able to synthesis all their organic compounds.
 Able to utilise atmospheric carbon dioxide and nitrogen.
 They are capable of independent existence in water and soil.
4. Heterotrophs
 Unable to synthesis all their own metabolites and depend on pre
formed organic compounds.
 Some organisms may require only a single organic substance like
glucose, while others may need a large amount of different
compounds like amino acids, nucleotides, lipids, carbohydrates and
coenzymes.

Bacteria require a supply of inorganic salts like sodium, potassium, magnesium etc.
which are normally present in the environment. Some bacteria require certain organic
compounds in minimum quantities. Growth does not occur in their absence, for some
bacteria these are accessory which enhance growth without being absolutely necessary
for them.
 FACTORS AFFECTING GROWTH OF MICROBES

1. Temperature requirements.
 Most bacteria grow best at moderate temperatures. These are called
mesophilic
 Cold loving bacteria thrive in temperature between 0° to 30°C. These
are called psychotrophs.
 Those that thrive in high temperatures between 40° C and 70° C are
called thermophilic.

Optimum temperature for most saprophytes is around 25°C and for most pathogen is
37° C.

2. Moisture and drying

Water is essential ingredient of bacterial protoplasm and hence drying is lethal
to bacterial cell like other living organisms.
But effect of drying varies in different organisms.
Eg. Treponema pallidum – highly sensitive.
Staphylococci - withstand drying for months.
Spores -resistant and may survive in the dry state for several
decades.
3. Food requirements.
Bacteria do not have chlorophyll so they cannot carry out photosynthesis.
 Few obtain energy from inorganic substances. These are termed as
autotrophic. Eg. Many soil bacteria.
 Majority derive energy from organic material, which are called as
heterotrophic.
 If they live on living organisms, they are called as parasites.
 If their food is from non living organic matter, they are called
saprophytes.
 COLLECTION OF SPECIMENS.

SPECIMEN

A specimen may be defined as a small quantity of a substance or object which shows

the kind and quality of the whole.

 For successful isolation and identification of pathogens, accuracy is required in

 Selection of specimen
 Collection time
 Storage
 Method of dispatch to laboratory.
 Aseptic handling during all steps of specimen collection.
 Precautions of specimen collection

 Specimen should be collected under strict aseptic conditions.

 Specimen should contain only those organisms from the site where it is
collected.
 Avoid contaminating discharges or ulcer material with skin commensals.
 Should be collected in dry, sterile, leak proof containers free from all traces
of disinfectants.
 Each specimen must be clearly labelled with patient’s name and
identification number.
 Every specimen should be accompanied by request form which includes
the information like patient’s name, date and time of collection, ward
details if it is collected in hospital, investigation required, clinical notes etc.
 High risk specimens like HIV, Hepatitis B must be handled with extra care.
For easy recognition of this type of specimens for taking extra care, the
words like HIGH RISK SPECIMEN should be mentioned on the label.

COLLECTION OF SPECIMENS - PURPOSES:

1 To detect any pathological changes

2 To identifies the infective organisms in different disease condition.
3 To conform the diagnosis.
 4 To monitor response to the treatment.
5 To identify the high risk cases.
6 To find out the possible complication.

TYPES OF SPECIMEN COLLECTION FOR INVESTIGATIONS:

1 Urine (mid stream)

2 Blood
3 Stool
4 Rectal swab
5 Sputum
6 Throat swab
7 Bone marrow
8 CSF (cerebrospinal fluid)
9 Body fluids (amniotic fluid, peritoneal fluid, synovial fluid, bile, pericardial
fluid, pleural fluid)
10 Skin scrapings

SPECIMEN COLLECTION OF URINE.

Indications for collection of urine:

1 Detection of uncontrolled Diabetics Mellitus as indicated by the presence of
glucose and ketones.
2 Detection of possible complication of pregnancy an indicated by proteinuria.
3 Detection of various types of renal diseases.
4 Detection of liver diseases.
5 Detection of obstruction within the biliary tract.
COLLECTING A CLEAN CATCH MID STREAM SPECIMEN

EQUIPMENT
1 Soap and water
2 Cleaning swab
3 Sterile specimen container (wide mouth bottle)
4 Label
5 Clean gloves.
PROCEDURE
1 Gather equipment
2 Wash the hands
3 Identify the client.
 4 Explain the procedure.
5 Provide perineal care as needed.
For male:
1. Cleanse the end of the penis with soap and water.
2. Collect mid stream urine after passing several amount of ml.

For female:
1. Clean the area around the urethral opening with water.
2. Hold labia apart and collect mid stream urine after passing several amounts of
ml.

Transport to laboratory: within 2 hours at 2-8° C.

Storage prior to processing: 24 hour at 2-8° C.

COLLECTING A 24 HOUR URINE SPECIMEN:

EQUIPMENT:
Specimen container.
Label
PROCEDURE
1. Explain the procedure to client
2. Collect urine specimen and discard it (first specimen is considered “old urine).
3. Record date and time of first specimen and label it.
4. Place all urine voided in specific container
5. Request client to void exactly 24 hrs , after first specimen was obtained place
voided urine in container.
6. After the last voided specimen is placed in a bottle , cover and send entire
specimen to the lab .

COLLECTING AN INDWELLING CATHETER SPECIMEN

EQUIPMENTS;

20 ml syringe
21 G -22 G needle
Tube clamp.
Sterile specimen bottle.
Gloves.
 Alcoholic sponge
Label.
Lab form.

STEPS OF PROCEDURE
1. About 30 min before collecting the specimen, clamp the drainage tube to
allow urine to accumulate.
2. Wash hand and put gloves.
3. If the drainage tube has a sampling port, aspirate the specimen into syringe.
4. If drainage tube does not have sampling port obtain specimen from catheter.
5. Transfer the specimen to a sterile container label it and send it into lab.
6. If the catheter is not made of rubber or has no sampling port, wipe the area
where the catheter joints with a drainage tube with alcoholic sponge.
7. Disconnect the catheter and allow urine to drain into the sterile specimen
container.
8. Avoid catheter touching inside the sterile container.
9. Wipe the container and cap it.

SPUTUM COLLECTION

INDICATION:

1 Differentiation of gram positive from gram negative bacteria.

2 Diagnosis of respiratory infection.
3 Confirmatory diagnosis of tuberculosis.
4 Monitoring the response to treatment of respiratory infection.

METHOD OF COLLECTION:

1. Explain the procedure to patient.

2. Wash hands.
3. Collect the following equipments.
 Sputum specimen container with lid - 1
 Label
 Tissues – 4-5 pieces
 Paper bag – 2
4. Tell the client to rinse the mouth before coughing.
5. Tell the client to take several deep breathing and to cough up sputum
directly into the container.
6. Obtain sputum in container and close and seal it.
7. If the client is unable to produce sputum assist the client by placing the
palms of the hands or a rolled pillow over the incisional area if pain.
 8. Evaluate the client status after procedure.
9. Deliver the sputum to lab within 30 min.

STOOL SPECIMEN

INDICATION
1 Determine the presence of blood, parasites, bile, pathogen or substance as
ingested drugs.
2 Gross examination of stool to determine the characteristics such as colour,
consistency and odour.
PROCEDURE:
1 Explain the procedure to client.
2 Before collecting the specimen, ask to the client to void and not to void on
specimen container.
3 Wear gloves.
4 Clean out all urine from bed pan.
5 Raise the head of the bed, so that the client assumes the squatting position on
bed pan.
6 Provide privacy while passing stool.
7 Remove the bed pan.
8 Use tongue blade to obtain small portion of stool in a plastic container.
9 Discard remaining stool and clean the bed pan.
10 Remove the gloves and wash hands.
11 Label it and send to lab immediately.
12 Maximum storage time prior to processing is 72 hours at 2-8º C.

COLLECTION OF VOMITUS:

VOMITUS:

It is the material, which is excreted through the forcible ejection of the contents of
the stomach.

COLLECTION OF SPECIMEN:

1 The vomitus should be collected within 48-72 hours of vomiting.

2 It should be collected in a sterile container.
3 It should be transported to lab as early as possible with label on it.

COLLECTION OF BLOOD CULTURE SPECIMEN:

 Prepare the patient same as venipucture.
 After withdrawing wipe the top of culture tube with spirit and change the
needle on the syringe, and use to withdraw the blood.
 Label the culture bottle and send it immediately.
  Transport to laboratory should be within 2 hours at room temperature.
 Storage prior to processing should be only incubation at 37°C.

CSF
 Collect CSF in a sterile dry container.
 Disinfect skin before aspirating CSF.
 Send immediately to lab after labeling.
 Storage prior to processing should be 6 hours at 37°C

BODY FLUIDS
 Include Amniotic, abdominal, synovial, bile, pericardial, pleural fluids.
 Disinfect skin before aspirating.
 Take specimen in a sterile container.
 Transport immediately to lab after labeling.

SKIN SCRAPINGS
 Take the specimen in a sterile container.
 Do not allow specimen to dry out, if necessary moisten with sterile distilled
water.
 Transport to lab within 24 hours at room temperature.

IMMUNITY
 Term
Resistance exhibited by the host towards injury caused by microorganisms.

TYPES OF IMMNUNITY
1. INNATE IMMUNITY

a) Non specific Species

Racial
b) Specific Individual

2. ACQUIRED IMMUNITY
Natural
a) Active
Artificial

Natural
b) Passive
Artificial

INNATE OR NATIVE IMMUNITY

Resistance to infection which an individual possesses by virtue of his genetic and

constitutional make up.
 Not affected by prior contact with micro-organisms or immunisation.
  May be nonspecific or specific.
 Non specific means a degree of resistance to infection in general.
 Specific means resistance to a particular pathogen.
 Innate immunity may be considered at the level of species, race or individual.

Species immunity
Refers to the total or relative refractoriness to a pathogen, shown by all members of a
species.
Eg. All human beings are totally unsusceptible to plant pathogens and to many animal
pathogens.
Racial immunity
Within a species different races may show differences in susceptibility to infections.
Eg. High resistance of Algerian sheep to anthrax.
People of Negroid origin are more susceptible than Caucasians to TB.
Individual immunity
Differences in innate immunity exhibited by different individuals in a race is known
as Individual immunity.
 It is mainly due to genetic basis.
 Eg. Homozygous twins exhibit similar degrees of resistance or susceptibility to
lepromatous leprosy and TB.

Factors influencing level of innate immunity in an individual.

1. Age
 Two extremes of life carry higher susceptibility to infectious diseases as
compared with adult.
 Fetus in utero is normally protected from maternal infection by the
placental barrier.
 Some pathogen cross this barrier causing infections resulting in fetal
death.
 Eg. Rubella, Herpes, Cytomegalo Virus, Toxoplasma gonadii lead to
congenital malformations.
 High risk for foetus to infection is related to immaturity of its immune
apparatus.
 Old persons are highly susceptible to infections due to waning of
immune responses and other infirmities like enlarged prostate leading to
urinary stasis.
 2. Hormonal influences
Endocrine disorders diabetics, hypothyroidism and adrenal function are
associated with an enhanced susceptibility to infections.
 High incidence of staphylococcal sepsis in diabetics may be related to
increased level of carbohydrates in tissues.
 Corticosteroids in our body depress host resistance by suppression of
antibody formation.
3. Nutrition
Both humoral and cell mediated immunity are lowered in malnutrition.

Mechanism of innate immunity

1. Epithelial surfaces
Intact skin and mucous membranes covering the body protect against invasion
of microorganisms.
2. Antibacterial substances in blood and tissues
3. Microbial antagonisms.
4. Cellular factors in innate immunity.
5. Inflammation
6. Fever
7. Acute phase proteins