MICROBIOLOGY.

OLD PAPERS 2 MARK QUESTION & ANSWERS.

2017(JAN) – 2002
D.N.S.SAMHARSHA
B.R.V.N. SAI MANOHAR

 LOUIS PASTEUR:
a) Father of microbiology.
b) Established that fermentation is caused by microbial agents.
Contributions:
1. Development of methods & techniques of bacteriology.
2. Proved all forms of life , even microbes arose from their like and not de novo.
3. Disproved spontaneous generation therory by swan neck flask experiments.
4. Introduction of sterilization techniques , development of steam sterilizer , autoclave & hot air owen.
5. Studies on anthrax , chicken cholera & hydrophobia.
6. Live Vaccine : introduced attenuated live vaccine for prophylactic use. Pasteur coined the term vaccine for prophylactic
preparations.

 ROBERT KOCH:
a) Father of bacteriology.
Contributions :
 Introduced methods for isolation of pure strains of bacteria.
 Introduced methods obtaining bacteria in pure culture using solid media.
 Introduced staining techniques.
 Dicovered anthrax bacillus , tubercle bacilli & cholera vibrio’s.
Koch’s postulates :
 Microorganism is accepted as causative agent of infectious disease , if following conditions are satisfied.
1. Organisms should constantly associated with lesions of diseasa.
2. It should be possible to isolate organism in pure culture from lesions of the disease.
3. The isolated organism (in pure culture) when inoculated in suitable lab animals s hould produce a similar
disease.
4. It should be possible to re isolate the organism in pure culture from lesions produced in experimental
conditions.

 NAME 4 STAINING TECHNIQUES IN MICROBIOLOGY:

1) Simple stains:
 Basic dyes such as methylene blue / basic fuscin are used as simple stains.They provide colour contrast but impart same
colour to all the bacteria in the smear.
2) Negative staining:
 Bacteria are mixed with dyes such as indian ink / nigrosin.The background gets stained and unstained bacteria stand out
in contrast.This is very useful in demonstration of bacterial capsules which do not take simple stains.
3) Impregnation methods:
 Bacterial cells and structures that are too thin to be seen under the light microscope , are thickened by impregnation of
silver on the surface to make them visible . , e. g: Demonstration of bacterial flagella and spirochates.
4) Differential stains:
 They impart different colours to different bacteria or bacterial structures. The two most commonly employed differential
stains are the Gram stain and the Acid – fast stain.

 NAME CAPSULATED BACTERIA:

1) Streptococcus pneumonia,
2) Klebsiella species . Bacillus anthracis,
3) Cryptococcus neoformans (fungus).

 DEMONSTRATION OF CAPSULE:
 A capsule is a gelatinous outer layer secreted by bacterial cell and that surrounds and adheres to the cell wall.
 Most capsules are composed of polysaccharides, but some are composed of polypeptides.
 The capsule differs from the slime layer that most bacterial cells produce in that it is a thick, detectable, discrete layer outside the
cell wall.
 The capsule has little affinityfor basic dyes,therefore,it can’t be stained by Gram staining.
 The following methods have been used for demonstration of capsule.
I. India Ink Staining(negative staining);
Capsule appears as a clear halo around bacterium as the ink can’t penetrate the capsule.
II. Serological methods;
When a Suspension of capsulated bacterium is mixed with its specific anticapsular serum and examined under the
microscope,the capsule appears ‘Swollen’ due to increase in its refractivity.
This phenomenon is called capsule swelling reaction (or) Quelling phenomenon(or) Neufeld reaction.

 INCLUSION BODIES:
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  These are sources of stored energy and present in some species of bacteria.
 These are membrane less.
 These are lie freely in cytoplasm.
 Their function and significance are uncertain.

 BACTERIAL FLAGELLA:
 Flagella are the complex filamentous cytoplasmic structure protruding through cell wall.
 These are unbranched, long, thread like structures, mostly composed of the protein flagellin, intricately embedded in the cell envelope.
 They are about 12-30 nm in diameter and 5-20 µm in length.
 They are responsible for the bacterial motility.
 Motility plays an important role in survival and the ability of certain bacteria to cause disease.
 There are 4 types of flagellar distribution on bacteria
1. Monotrichous
– Single polar flagellum
– Example: Vibrio cholerae
2. Amphitrichous
– Single flagellum on both sides
– Example: Alkaligens faecalis
3. Lophotrichous
– Tufts of flagella at one or both sides
– Example: Spirillum
4. Peritrichous
– Numerous falgella all over the bacterial body

– Example: Salmonella Typhi

 Each flagellum consists of three distinct parts- Filament, Hook and Basal Body.
 The filament lies external to the cell.
 Hook is embedded in the cell envelope.
 Basal Body is attached to the cytoplasmic membrane by ring-like structures.

 Funtions of flagella are ;

 Movements
 Sensation
 Signal transduction
 Adhesion

 TRANSPORT MEDIA:
 These are used in the case of delicate organisms (e.g ; gonococci) which may not survive the time taken for the transit or may be
overgrown by the non – pathogenic bacteria (e.g ; cholera organisms).
 For transport of specimens to the laboratory , special media are devised and these are termed transport media.
Examples of transport media are:

a) Stuart’s transport medium is a non – nutrient soft agar gel containing a reducing agent to prevent oxidation , and charcoal to
neutralise bacterial inhibitors.It may be used for organisms such as gonococci.
b) Buffered glycerol saline transport medium for enteric bacilli.

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 NAME 4 ANAEROBIC BACTERIA:
1) Peptostreptococci
2) Actinomycetes
3) E.coli
4) Klebsiella pneumonia.

 DIFFERENT MORPHOLOGICAL FORMS OF BACTERIA:

Depending upon shapes ;
a) Cocci: oval / spherical cells
In pairs – Diplococci
In chains – Streptococci
In clusters – staphylococci.
b) Bacilli : rod shaaped cells ., some have peculiar arrangement or shape.
i. Coccobacilli . , e.g ; Brucella.
ii. Streptococci : arranged in chain . , e.g ; Streptobacillus.
iii. Chinese letter / cunie form ., e.g ; corynebacterium.
iv. Comma shaped . , e.g ; vibrio
v. Spirilla ., rigid spiral forms. E.g ; spirellum.
c) Spirochaetes : slender flexous spiral forms . , e.g ; Treponema.
d) Actinomycetes : branching filamentous
Bacteria rembling fungi.
e) Mycoplasms : cell wall deficient
Do not posses a stable shape.

 DRAW A DIAGRAM OF SPORE AND LABEL :

 GROWTH CURVE:

 When bacterium is inoculated into suitable culture media is incubated , growth follows definite course.
a) Lag phase:

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 Period between inoculation & beginning of multiplication is called Lag phase.There is increase in size of the cells , no
appreciable increase in numbers.
b) Log (Logarithm) or Exponential phase :
Cell division starts & their number increases.,Exponentially or by geometric progression with time.
c) Stationary phase:
Bacterial growth ceases almost completely due to exhaustion of nutrients and accumulation of toxic products .,number of vaible
cells remains stationary as there is balance between dying cells & newly formed cells.
d) Phase of decline :
Bacterial cell population decreases due to death of cells. This phase starts due to exhaustion of nutreints accumulation of toxic
products and autolytic enzymes .There is decline in viable count not total count.

 USES OF HOT AIR OVEN:

 It is used for sterilisation of ;
i. Glass – wares like glass syringes , petridishes , flasks , pipettes and test tubes.
ii. Sugical instruments like scapels , scissors , forceps etc.
iii. Chemicals such as liquid paraffin , fats , sulphonamide powders etc.

 PRINCIPLE OF AUTOCLAVE:
 Steam above 100˚ C or saturated steam has better killing power than dry heat . Bacteria are more susceptible to moist heat as
bacterial protein coagulates rapidly.
 When steam comes in contact with cooler surface it condenses to water andliberates its latent heat to that suface.
 This process continues till the temperature of that article is raised to that of steam . The condensed water produces moist
conditions for the killing microbes present.

 NAME 2/3 ENRICHED (2) AND 2 TRANSPORT MEDIA AND USES(1):

ENRICHED MEDIA :
 When basal medium is added with some nutrients such as blood , serum or egg , it is called Enriched medium.Some examples of
enriched media are:
1) Blood agar – Blood is added to nutrient agar.It may be used for growing a number of bacteria but one specific example is
Streptococcus which requires blood for its growth.
2) Chocolate agar – It is heated blood agar used for isolation of Neisseria and Haemophilus influenzae.
3) Loeffler’s serum slope – Serum is added for enriching the medium . The medium is used for isolation of Corynebacterium
diphtheriae.
TRANSPORT MEDIA:
 These are used in the case of delicate organisms (e.g ; gonococci) which may not be survive the time taken for transit or may be
overgrown by non – pathogenic bacteria (e.g ; cholera organisms) .
 For transport of specimens to the laboratory, special media are devised and these are termed Transport Media.examples are:
1) Stuart’s transport medium is a non – nutrient soft agar gel containing a reducing agent to prevent oxidation , and charcoal
to neutralise bacterial inhibitors.It may be used for organisms such as gonococci.
2) Buffered glycerol saline transport medium for enteric bacilli.

 MENTION 4 SELECTIVE MEDIA(4)AND USES:

1) Selective media contain substances that inhibit all but few types of bacteria & facilitate the isolation of particular specie s.
2) This is used to isolate particular bacteria from species where mixed bacteria flora is expected.
Examples:
1. Deoxycholate citrate agar (DCA) – Addition of DC acts as selective agent for enteric bacilli (Salmonella , Shigella).
2. Bile salt agar (BSA) – bile salts is added as selective agent .It favours growth of the vibrio cholerae.

 EXOTOXIN :
1) Thet are heat labile proteins which a secreted by certain species of bacteria.
2) Diffuse readily to surrounding media.
3) Highly potent even in minute amounts.
4) When treated with formaldehydes, coverted to toxoids .These toxoids lack toxicity but retain antigenicity thus induce protective
immunity whrn used as vaccines.
e.g ; Tetanus toxoid.
5) Highly antigenic and stimulate formation of antitoxin ehich neutralises toxin.
6) Highly specific for particular disease.
e.g. ; Tetanus for CNS.
7) Have specific pharmological activities and donot produce fever in the host.
8) Mainly produced by gram +ve , may also by gram –ve (e.g ; Vibrio cholerae) and enterotoxigenic E.coli

 DEFINE ANTIGEN:
Antigen is a substance introduced into the body evokes immune response to produce a specific antibody with which it reacts in a observable
manner.
Types:
1. Complete antigen,
2. Haptens / incomplete antigens : Complex haptens , Simple haptens

 FOUR FUNCTIONS OF Ig G:
1. Ig G is the major serum immunoglobulin (80% of total amount) .Normal serum concentration – 8 – 16 mg/ml.
2. Only immunoglobulin that is transported through placenta and provides natural passive immunity to the newborn.

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 3. It appears late but persists for longer period.It appears after initial immune response which is ig M in nature.
4. It participates in precipitation , complement fixation and neutralisation of toxin & virus.
5. It is protective against those microorganisms which are active in the blood and tissues.

 NAME AGGLUTINATION TESTS USED FOR DIAGNOSIS :

1) Slide agglutination test,
2) Tube agglutination test,
3) Coomb’s test(anti globulin test)
4) Heterophilic agglutination test
5) Passive agglutination test
i. Latex agglutination test
ii. Haemoagglutination test
iii. Coagglutination.

 WRITE 4 DIFFERENCES B/W EXOTOXIN AND ENDOTOXIN(2):

EXOTOXINS ENDOTOXINS

1) Heat labile (>60ºC) 1) Heat stable.

2) Protein M.W 10,000 – 9,00,000 2) Lipopolysaccharide in nature.
3) Converted to toxoid by formaldehyde. 3) Cannot be toxoided.
4) Enzyme in action. 4) No enzyme action.
5) Very high potency. 5) Low potency.
6) Do not produce fever in host. 6) Usually produce fever.
7) Highly specific. 7) Non – specific.
8) Specific pharmocological effect for each 8) Non – specific action of all endotoxins.
exotoxin.
9) Highly antigenic. 9) Weakly antigenic.
10) Produced by gram +ve also by some 10) Produced by gram – ve bacteria.
gram –ve bacteria.

 FOUR MODES OF TRANSMISSION OF INFECTION:

a) Contact:
1. Direct – STD’s –syphilis , gonorrhoea , AIDS are acquired by direct contact.
2. Indirect – May be through agency of formites, such as clothing , toys etc.,that are contaminated by pathogen and act as
vehicle for transmission.
b) Inhalation:
 Respiratory infections such as common cold , influenza , whooping cough and tuberculosis are acquired by inhalation.
 These organisms are shed into environment by patients in secretion from nose / throat during sneezing , coughing or
speaking.
c) Ingestion :
 Intestinal infections like cholerae , dysentry , food poisioning and most of the parasitic infections are acquired by ingestion
of food or drink contaminated by pathogenes.
d) Vectors:
 Vectors are arthropods or other invertebrate hosts.
 E.g ; mosquitoes , flies , ticks , mites , and lice.Transmission by vector may be either mechanical or biological.
e) Inoculation:
 Pathogenes in some instances, may be inoculated directly into tissue of the host , for example Rabies virus is inoculated
directly by bite of rabid animal.
f) Transplacental
g) Iatrogenic and laboratory infections.

 TYPES OF ACTIVE IMMUNITY WITH EXAMPLES :

Active immunity is subdivided into 2 types:
1) Natural – through clinical / sub clinical infection.
2) Artificial – induced by vaccination.
NATURAL ARTIFICIAL
 Acquired by natural clinical / sub clinical  Inducced by vaccination prepared from live ,
infections. attenuated or killed Microorganisms.
 Long lasting  E.g ; Live vaccine
 Persons recovering from small pox development BCG for tuberculosis,
of natural active immunity. Sabin vaccine for poliomyelitis.
Killed vaccine
Hepatitis B vaccine,
Killed cholerae vaccine,
TAB for enteric fever,
Bacterial products
Tetanus toxoid – Tetanus,
Diptheria toxoid – Diptheria.

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 FOUR DIFFERENCES BETWEEN ACTIVE AND PASSIVE IMMUNITY :
ACTIVE IMMUNITY PASSIVE IMMUNITY
1) Produced actively by immune system. 1) Received passively by the host.The host’s immune
system does not participate.
2) Induced by infection / by contact with immunogens. 2) Conferred by administration of ready – made
antibodies.
3) Long-lasting and effective protection . 3) Protection short lived & less effective.
4) Not applied in immuno deficient persons. 4) Applicable in immunodeficient persons.
5) Used for prophylaxis to increase body resistance e.g ; 5) Used for treatment of acute infections.
BCG vaccine.
6) Negative phase may occur 6) No negative phase.
7) Immunological memory present 7) No immunological memory.

 IgA:
 It is the second major serum immunoglobulin . The normal serum concentration is 0.6 – 4.2 mg / ml.
 Half – life is about 6 -8 days.
 It occurs in two forms serum Ig A.,and secretory Ig A.
 Serum Ig A is monomeric 7S molecule ,while Ig A found on mucosal surfaces and in secretions is a diamer formed by two monomer
units joined together by a glycoprotein named the J chain (J for joining).
 Secretory Ig A contains another polypeptide called secretory piece or secretory component. The S piece is believed to protect IgA
from denaturation by bacterial proteases in sites such as the intestinal mucosa while is rich in bacterial flore.
 These are present in secretions such as milk , saliva , tears , sweat , nasal fluids , colostrum and in secretions of respiratory ,
intestinal and genital systems.It protects the mucous membranes against microorganisms.

 SECRETORY IgA:
 It is dimer formed by 2 monomeric units joined together by glycoprotein named the J chain.
 It contains another polypeptide chain called secretory piece / secretory component.
 The S piece is believed to protect ig A from denaturation by bacterial proteases in the sites such as intestinal mucosa whic h is rich
in bacterial flora.
 Ig A is principle ig present in secretions such as milk , saliva , tears , sweat , nasal fluids etc.,
 It is synthesized locally by plasma cells and little serived from serum.

 DRAW A NEAT LABELLED DIAGRAM OF Ig A ANTIBODY :

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 TYPE III HYPERSENSITIVITY:
 Also called immune complex reaction.
 It is characterised by deposition of Ag – Ab complexes in tissues , activation of complement and infiltration of polymorphonuclear
leucocytes leading to tissue damage.
 It includes
i. Arthus reaction,
ii. Serum sickness
Arthus reaction Serum sickness
 Localised  Generalised
 Due to relative antibody excess.  Due to relative antigen excess.
e.g ; post streptococcal glomerulonephritis ,
dengue haemorrhagic fever and malaria.

 ENUMERATE ANTIGEN ANTIBODY REACTIONS:

 Antigen – antibody reactions Invitro are called serological reactions.They are useful in laboratory diagnosis of various diseases and
in identification of infectious agents.
 Various antigen – antibosy reactions are as follows:
1) Precipitaion:
 When soluble antigen reacts with antibody in presence of electrolytes at optimum pH & temperature , the
antigen – antibody complex forms an insoluble precipitate , it is called precipitation.
e.g ; VDRL test for syphilis.
2) Agglutination:
 Ag – Ab reactions , particular Ag combines with its Ab in the presence of electrolytes , at optimum temperature ,
pH resulting in visible clumping of particles.
e.g ; WIDAL test for typhoid , Coomb’s test for detection of incomplete antibodies , Haemoagglutination testfor blood
grouping & typing.
3) Complement fixation:
 Test for kala azar , amoebiasis , viral disease.
4) Neutralisation test:
 Toxigenic test for C.diptheriae,
 Shick test – Diptheriae,
 Virus neutralisation tests.
5) Immunoflourescence
 Direct and Indirect
6) RIA(radio immunoassay)
7) ELISA
8) Immunochromatography.

 VDRL TEST:
 It is veneral diseases research laboratory test.
 It is a serological test and non treponemal test used for laboratory diagnosis of syphilis.
 Veneral disease research laboratory test.
 Most widely used simple & rapid test . It is a slide flocculation test.,The VDRL antigen (cardiolipin) must be prepared fresh dialy.

 COAGULASE TEST:
 Coagulase converts fibrinogen into fibrin.
 The test for coagulase is done by the Slide and the Tube method .It is done to differentiate pathogenic(Staph.aureus) strain from
non – pathogenic strains.
i. Slide coagulase test :-
 It detects bound coagulase.
 A few colonies of bacteria are emulsified in a drop of normal saline on a clean glass slide and mixed with a drop
of undiluted rabbit or human plasma.
 More clumping of the suspension occurs with coagulase positive strains.

ii. Tube coagulase test :-

 It is done for detection of extracellular free coagulase.
 0.1 ml of an overnight broth culture (or) an agar culture suspension of the organism is mixed with 0.5 ml of a 1
in 5 dilution of human or rabbit plasma.
 Diluted plasma alone in a second tube serves as a control.The tubes are incubated in water bath for 3-6 hours
at 37ºC.
 In case of a positive test , the plasma clots and does not flow when the tube is inverted.

 DEFINE DEFINITIVE HOST WITH EXAMPLES:

 It is the host that harbours the adult stage of the parasite or where the parasite replicates sexually .
 Example ,
1. For plasmodium vivax definitive host is Female anopheles mosquito.
2. For taenia solium definitive host is man.

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 3. For Wuchereria bancrofti definitive host is man.

 DEFINE INTERMEDIATE HOST WITH EXAMPLES :

 It is the host that harbours the larval stages of the parasite or where the parasite replicates asexually.
 Examples,
1. For plasmodium vivax intermediate host is man.
2. For taenia solium intermediate host is pig.
3. For wuchereria bancrofti intermediate host is female culex mosquito.

 HIV :
 The human immunodeficiency virus (HIV) , the causative agent of AIDS , belongs to retrovirus.
 HIV is a spherical enveloped virus containing two identical copies of single stranded genome.In association with viral RNA is the
reverse transcriptase enzyme. The virus core is surrounded by a nucleocapsid composed of protein.The virus contains a lipoprotein
envelope.
 There are three modes of transmission of HIV infection ., These are sexual , contact , parental and perinatal.
 ELISA is the method most commonly used test.
 Postexposure prophylaxis (PEP) may be required when there is exposure to blood , body fluids , other potentially infected material
or an instrument contaminated with HIV.

 WRITE A NEAT LABELLED DIAGRAM OF HIV(1) / DIAGRAM OF AIDS VIRUS(1):

 ROUTES OF TRANSMISSION OF HIV INFECTION(1) / MENTION 4 MODES OF INFECTION OF HIV(2):

 Sexual contact : This is the most important mode of transmission . Sexual transmission occurs among both homosexual as well a s
heterosexual individuals.
 Parental transmission : It may occur through blood after receiving infected blood transfusions , blood products , sharing
contaminated syringes and needles as in intravenous drug abusers or accidental inoculation.
 Perinatal transmission : Infection may be transmitted from an infected mother to her child either transplacentally or perinatally.

 MENTION FOUR OPPORTUNISTIC INFECTIONS IN HIV:

I. Bacterial
1. Mycobacterial infections – Tuberculosis & non – tuberculosis infections.
2. Salmonellosis.
II. Viral
1. Herpes simplex
2. CMV(cytomegalo virus)
3. Varicella – zoster
III. Mycotic
1. Candidiasis
2. Histoplasmosis
3. Cryptococcosis
4. Coccidiodomycosis
IV. Parasitic
1. Toxoplasmosis
2. Isosporiasis
3. Cryptosporidiosis
V. Malignancies
1. B – cell lymphoma or non – hodgkin’s lymphoma.
2. Kaposi’s sarcoma.

 POSTEXPOSURE PROPHYLAXIS (PEP) FOR HIV:

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  Exposure to blood,body fluids,other potentially infected material or an instrument contaminated with one of these materials may lead
to risk of acquiring HIV infection.
 The risk of infection varies with type of exposure and other factors.
 PEP (post-exposure prophylaxis) means taking antiretroviral medicines (ART) after being potentially exposed to HIV to prevent
becoming infected.
 It is depending upon the category of exposure and HIV sataus of exposure source.,Zidovudine 300 mg BD and Lamivudine 150 mg
BD are used in basic two drug regimen.
 The drugs must be started within the first 72 hours and ideally within 2 hours..,and continued for a period of four weeks.
 Besides PEP ,injured site on the wound should be thoroughly washed with soap and water.Antiseptics may also be used.

 NAME FOUR DNA VIRUSES:

 Vaccina virus.
 Cowpox virus.
 Herpes simplex virus.
 Varicella – zoster virus.
 Variola virus.

 NAME 3 RNA VIRUSES:

 Polio virus
 Rhabdo virus
 Rota virus
 Paramyxovirus.

 TWO IMPORTANT METHODS FOR DIAGNOSING AIDS INFECTION:

I. Specific tests for HIV infections
1. Antigen detection – ELISA.
2. Virus isolation – Important test for diagnosis in window period when antibodies are absent in serum of patients.
3. Detection of viral nucleic acid – PCR.
4. Antibody detection –
Screening tests – ELISA , Rapid tests , Simple tests.
Supplemental tests – Western blot test, Indirect immunoflourescent test.
II. Non – specific tests
1. Total and differential leucocytic count.
2. T lymphocyte subset assays.

 4 MARKERS OF HEPATITIS B VIRUS :

Marker Definition Significance
Hbs AG HBV surface antigen Positive in most cases of acute or chronic
infections.
Hbe AG E antigen ; a component of HBV core Transiently positive in acute hepatits B,
May persist in chronic infection
Reflects presence of viral replication , whole
dane particles in serum and high infectivity.
Anti – Hbc Antibody to C Antigen Transiently positive in convalescence
Maybe persistently present in chronic cases.
Usually reflects low infectivity.
Anti – Hbc (Ig M or Ig G) Antibody to HBV core antigen Positive in all acute and chronic cases.
Ig M anti – Hbc reflectts active viral
replication
Not protective

Anti – Hbs Antibody to HBV surface antigen Positive in late covalscence in most acute
cases confers immunity.

 HERPES SIMPLEX VIRUS INFECTIONS(1)&NAME 3 LESIONS:

 Man is the only natural host.There are two types of Herpes simplex virus , type 1 and type 2.
 HSV type -1 scientifically named Human herpes virus 1 (HHV -1 )
 HSV type -2 scientifically named Human herpes virus 2 (HHV -2)
 HSV – 1 infections :
 Herpes labialis,
 Keratoconjunctivitis,
 Encephalitis,
 Dendritic keratitis.
 HSV – 2 infections:
 Genital herpes(penis , urethra , cervix , vulva , vagina )
 Neonatal hepes
 Aseptic meningitis.
 HSV – 1 is primarily associated with lesions of the mouth , face , eyes and cns.
 HSV – 2 is primarily associated with lesions of the anogenital region , although both the viruses can infect any area.

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 MICROBIAL FLORA IN DENTAL PLAQUE:
 Streptococcus mutants and Streptococcus sanguis initially colonise the dental pellicle through adhesins.
 Later adiitional plaque area is such as actinomycetes can also bind.This may be followed by colonisation by other bacteria
prevotella intermedia.,Fusobacterium nucleatum., Porphyromonaas gingivalis.
 Other bacteria may be found are Mycoplasma species ., yeast & virus.
 If plaque calcifies , it is reffered to as ‘Calculus’ or ‘Tartar’.
 1 gram dental plaque = 2 x 1011 bacteria.
 In addition , It may also contain Epithelial cells , leukocytes , macrophages.

 DEFINE SEPTICAEMIA AND BACTERAEMIA:

 Septicaemia : The condition in which bacteria circulate and actively multiply in the blood stream.
Example:- salmonella typhi , nesseria meningitis.
 Bacteraemia: presence of bacteria in blood without any multiplication.

 BACTERIA CAUSING MENINGITIS(2)&MENTION FOUR BACTERIA(2):

I. Children &adults
1) Neisseria meningitis ( meningococcus)
2) Streptococcus pneumonia (pneumococcus)
3) Haemophilus influenzae
4) Staphylococcus aureus
5) Listeria monocytogenes
II. Neonates&infants
1) E.coli
2) Group B streptococci
3) Staph.aureus
4) H.influenzae
5) Strep.Pneumonia
6) Klebsiella species

 NAME 4 TAPE WORMS:

1. Taenia solium
2. Taenia saginata
3. Echinococcus granulosus
4. Echinococcus multilocularis
5. Bertiella mucronata
6. Bertiella studeri

 PROPHYLAXIS OF TETANUS :
 Available methods are;
a) Sugical methods – Depending on type of wound , may vary from simple cleansing to radical excision.
b) Antibiotics – long acting pencillin ijection or erythromycin.
c) Immunisation :
I. Active – most effective.
Tetanus toxoid available as plain toxoid or absorbed on aluminium hydroxide or phosphate (APT) commonly used.
Given along with DPT in children.
II. Passive – anti tetanus serum (ATS) prepared by immunising horses with toxoid given 1500 IU .,i.m Immediately after
wounding ,it produces hypersensitivity reactions.
Homologous serum prepared from humans ,human antitetanus immunoglobulins(HTIG)250 units.
III. Combined prophylaxis – In non – immune person , At one site immune with first dose of tetanus toxoid,
Another site immune with ATS or HTIG.,
Followed by 2 nd & 3rd doses of tetanus toxoid at monthly interval.

 CASTANEDO’S METHOD OF CULTURE

 The need of frequent subcultures can be avoided by the use of Castanedo’s method of culture.
 It contains both liquid(trypticase soy bath) and solid(trypticase soy agar) medium in the same bottle.
 The blood is inoculated into the broth (liquid medium) and the bottle is inoculated in upright position .
 For subculture the bottle is tilted so that the broth flows over the surface of solid agar slant.It is again inc ubated in the unright
position.
 In case of positive blood culture , colonies appear on the agar slant.
 This method reduces the chances of contamination and risk of infection to laboratory workers.

 MENTION 4 IMPORTANT BACTERIA CAUSING URINARY TRACT INFECTION AND ALSO WRITE WHAT IS
SIGNIFICANT BACTERIURIA:
I. Gram –ve bacilli
Most common infecting agents
1) E.coli – commenest cause of urinary tract infections .,responsible for 70 – 80% of infections in general populations.
2) Klebsiella species,
3) Proteus species .,especially P.mirabilis,
4) Enterobacter,

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 5) Pseudomonas aeruginosa
6) Serratia
II. Gram +ve cocci
1) Enterococci e.g ; E.faecalis,
2) Staph.saprophyticus,
3) Staph.aureus,
4) Staph.epidermidis – It is found in urine as a contaminant but may occur as pathogen in immunocompromised
individuals.
III. Miscellaneous
1) M.tuberculosis,
2) Citrobacter,
3) Salmonellae,
4) Gardnerella vaginalis,
5) Str.pyogenes,
6) Str.agalactiae.
 Significant bacteriuria : when bacteria count is more than 10 5 / ml of a single species.

 KOPLIC SPORTS OR BACTERIA IN ORAL CAVITY:

Gram positive Gram negative
Cocci Streptococcus mutants Veillonella species
Strept. sanguis Neisseria species
Salivarius Branhamella species
Mitior
Milleri
Enterococcus species
Peptostreptococcus species
Micrococcus species
Staphylococcus species.
Bacilli Lactobacillus species Actinobacillus species
Actinomycetes species Fusobacterium species
Propionibacterium species Bacteriodes species
Arachnia species Leptotricha species
Bifidobacterium species Eikenella species
Bacterionema species Selenomonas species
Eubacterium species Wollinella species
Troponema species
Haemophilus species

 LABORATORY DIAGNOSIS OF MALARIA:

1. DNA probes – sensitive & specific diagnostic methods for diagnosis of malaria.
2. Serological tests – indirect immunoflourescent test , Indirect haemoagglutination assay (IHA) , ELISA.
3. Microscopic examination – peripheral blood smear is most diagnostic (examination) procedure in malaria.Malarial parasit es quickly
detected but species identification is difficult.
Examination Organisms
All sexual erythrocytic stages gametocytes P.Vivax , malariae , Ovale , falciparum.
Multiple rings in individual RBC with accole forms P.falciparum
Schuffner’s dots (RBC) P.vivax , P.ovale
Maurer’s dots P.falciparum
Ziemans dots P.malariae
RBC enlarged P.vivax
RBC not enlarged P.falciparum

 MENTION 4 LIVE VACCINES(1) / NAME FOUR VACCINES(1) :

a) Live vaccines
 BCG for tuberculosis
 Sabins vaccine for poliomyelitis
 MMR vacccine – measels , mumps , rubella.
b) Killed vaccine
 TAB for enteric fever
 Neural & Non – Neural vaccines for rabies.
 Hepatits B vaccines
 Salk vaccine for poliomyeletis.
 Killed cholera vaccine
c) Bacterial products
 Tetanus toxoid for tetanus
 Dipthera toxoid for diptheria.

 MMR VACCINE:
 It is a combined Measles – mumps – rubella (MMR) vaccine.
 It is recommended for all infants at the age of 15 months , followed bya booster at the age of 4 – 6 years.

11
  it is also recommended for use in some cases of adults with HIV.

 MENTION 2 IMPORTANT LIVE VIRAL VACCINES:

 Live vaccines
 BCG for tuberculosis
 Sabins vaccine for poliomyelitis
 MMR vacccine – measels , mumps , rubella.
 Rota virus vaccine
 Rabies vaccine
 Chicken pox vaccine
 Small pox vaccine

 DPT VACCINE:
 DPT refers to a class of combination vaccines against three infectious diseases in humans: diphtheria, pertussis (whooping cough),
and tetanus.
 The vaccine components include diphtheria and tetanus toxoids and killed whole cells of the organism that cause pertussis .
 Three doses are given by intramuscular route at an interval of 4 – 6 weeks.Booster doses are given at 18 months and at 5 years.
 It is the one of the most common vaccine preventable childhood disease.

 BCG VACCINE:
1. Calmette and guerin prepared an attenuated strain of M.Bovis by growing it on potato medium.The strain is attenuated by repeated
subcultures.When the strain becomes incapable of producing tuberculosis in a susceptable Guinea pig.
2. BCG strains grow well on glycerol containing medium.
3. They are aerobic and resistant to cycloserine.
4. BCG vaccine is available in liquid & freeze dried (lyophilised) form.The freeze dried vaccine is reconstituted by sterile physiological
saline to make a concentrarion of 0.1 mg in 0.1ml vaccine.It should be utilised in 3 – 6 hours.
5. Vaccine is given intradermally in dose of 0.1ml
6. It is given soon after birth / any time during first year.
7. A small nodule develops at the site of infection , 2- 3 weeks after injection increases slowly and attains diameter 4 – 8 mm after
about 5 weeks.It breaks into shallow ulcer heals spontaneously leaving a 4 – 8 mm diameter permanent round scar.such individuals
become tuberculine +ve in 4 – 6 weeks.
8. Immunity is reported as last for about 10 years.
9. It is believed to protect severe serious forms of TB such as meningeal , skeletal & miliary forms of disease.
10. Contraindicated in patients of AIDS , Eczema , Pertusis , Measels and patients on steroids.

 WRITE ABOUT OPPORTUNISTIC FUNGI CAUSING INFECTION(1)NAME 3 (1) /

MENTION THE OPPORTUNISTIC FUNGAL INFECTIONS(1) :
 Some fungi usually donot produce disease but may cause infection under special conditions.The incidence of this infections is
increased due to widespread use of antibiotics , corticosteroids and immunosuppressive drugs.They are called opportunistic fungi.
 Aspergillus and pencillin grow on damp bread and other organic matter.
 The fungi can produce serious & even total infections in persons who are otherwise debilitated
 Candida albicans ,Aspergillus fumigatus , Aspergillus niger , Penicillium species , Rhizopus and Mucor are some examples of
opportunistic fungi.

 MENTION 4 SEXUALLY TRANSMITTED INFECTIONS/BACTERIAL DISEASES(3):

Painless Genital ulcers.
 Syphilis  Treponema pallidum
 Lymphogranuloma venerum (LGV)  Chlamydia trachomatis.
 Donovanosis  Klebsiella granulomatis

Painful genital ulcers.

 Chancroid  Haemophilus ducreyi
 Herpes genitalis  HSV – 1,2

Urethral discharge.
 Gonorrhaea  Neisseria gonorrhea.
 Non – gonococcal urethritis  Chlamydia trichomatis
Ureaplasma urealyticum
Mycoplasm genitalium
M.hominis
Vaginal discharge.
 Gonorrhaea  Neisseria gonorrhea.
 Trichomoniasis  T.vaginalis.
 Vaginitis  Gardnerella vaginalis
Mobiluncus species.
 Vulvovaginal candidiasis  Candida albicans.

Genital warts. Human papilloma virus.

No genital lesions but only systemic manifestations. HIV – 1 & HIV – 2 ,HBV , HCV.
Miscellaneous  Group B strepto cocci
 Cytomegalo virus

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  Shigella species
 Camplobacter species
 Giardia lamblia
 E.hystolytica.

 Bacterial sexually transmitted list:

 DIFFERENT TYPES OF SPORES:

a) Sexual spore:
4 types – Oospore,
Ascospore,
Zygospore,
Basidiospore.
b) Vegetative spore:
Blastospore – formed by budding from parent cell , as in yeasts.
Arthrospores
Chlamydospores.
c) Aerial spores:
Conidiospores – spores borne externally on sides or tips of hyphae.
Microconidia – conidia are small and single.
Sporangiospores – spores formed within sporangium e.g ; mucor , rhizopus.
Macroconidia – large & septate conidia & are often multicellular.

 ENUMERATE 4 NON SPORING ANAEROBES :

A. Cocci ;
Gram +ve ;
 Peptococcus
 Peptostreptococcus
Gram –ve ;
 Veillonella.
B. Bacilli ;
Gram +ve;
 Eubacterium
 Lactobacillus
 Bifidobacterium
 Propionibacterium
 Actinomycetes , Mobiluncus.
Gram –ve ;
 Bacteroides

13
  Prevotella
 Fusobacterium
 Leptotricha
C. Spirochaetes ;
 Troponema
 Borrelia.

 NAME 4 DISEASES CAUSED BY STAPHYLOCOCCUS AUREUS(1) / 4 INFECTIONS CAUSED BY

STAPHYLOCOCCUS(1) :
Staphylococcal diseases are classified as
A. Cutaneous infections.
 Superficial infections include boils , carbuncles , pustules , abscesss , styes , impetigo , pemphigus , neonatorum , wound and
burn infections.
B. Deep infections.
 Osteomyelitis , tonsillitis , pharyngitis , sinusitis , pneumonitis , emphyma , endocarditis , meningitis , bacteriameia septicemia ,
and pyaemia.
C. Food poisioning.
 Staphylococcal food poisoning may follow 2 – 6 hrs after ingestion of contaminated food which contains performed enterotoxin
of staphylococcus aureus.
D. Nosoconial infections.
 They are important cause of hospital acquired infections.
E. Skin exofoliative diseases.
 Staphylococcal scaled skin syndrome (SSSS) is one example of exfoliative disease in which toxin spreads systematically.It is
seen but not exclusively in small children.
F. Toxic shock syndrome (TSS)
 Caused by TSST – 1(Toxin shock syndrome toxin) characterised by high fever , hypotension , vomiting , diahhorea and
scarlatiniform rash.

 LESIONS CAUSED BY STAPHYLOCOCCUS AUREUS :

s.aureus is an important pyrogenic organism nad lesions are organised in contrast to streptococcal lesions which are spreading in nature.
 Coagulase enhances virulence of s.aureus by inhibiting phagocytosis.
 It forms wall of fibrin clot around the lesion thick creamy pus is formed in staphylococcal infections.
Staphylococcal diseases are classified as
1) Cutaneous infections.
 Superficial infections include boils , carbuncles , pustules , abscesss , styes , impetigo , pemphigus , neonatorum , wound a nd
burn infections.
2) Deep infections.
 Osteomyelitis , tonsillitis , pharyngitis , sinusitis , pneumonitis , emphyma , endocarditis , meningitis , bacteriameia septicemia ,
and pyaemia.
3) Food poisioning.
 Staphylococcal food poisoning may follow 2 – 6 hrs after ingestion of contaminated food which contains performed enterotoxin
of staphylococcus aureus.
4) Nosoconial infections.
 They are important cause of hospital acquired infections.
5) Skin exofoliative diseases.
 Staphylococcal scaled skin syndrome (SSSS) is one example of exfoliative disease in which toxin spreads systematically.It is
seen but not exclusively in small children.
6) Toxic shock syndrome (TSS)
 Caused by TSST – 1(Toxin shock syndrome toxin) characterised by high fever , hypotension , vomiting , diahhorea and
scarlatiniform rash.

 NAME 4 DISEASES CAUSED BY STREPTOCOCCUS PYOGENS

Pyrogenic infections include
a) Respiratory infections – sore throat (acute tonsillitis / pharyngitis).
b) Skin infections – lymphangitis & cellulitis.
c) Streptococcal toxic shock syndrome (TSS)
d) Pereperal sepsis, sepsis , pyaemia , septicemia abscess in internal organs.

 MENTION 3 DIFFERENCES B/W STREPTOCOCCAL VIRIDANS AND PNEUMOCOCCI:

Strptococcal viridians Streptococcal pneumococci

1. Morphology Capsulated,Lanceolate diplococci. Non – capsulated , oval / round cocci in
chains.
2. Colonies in blood agar Intially dome shape with α – haemolysis Dome shaped with α – haemolysis.
, later draughtman colonies.
3. Colonies in liquid medium Uniform turbidity. Granulat turbidity , powdery deposit.
4. Bile solubility
5. Inulin fermentation +ve -ve
6. Optochin sensitivity
7. Animal pathogenicity Fatal infection Non pathogenic.

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 MENTION 2 IMPORTANT NON – SUPPURATIVE COMPLICATIONS OF POST STREPTOCOCCAL PYOGENIC
INFECTION(1) / NON SUPPURATIVE LESIONS,CAUSED BY STREPTOCOCCUS PYOGENES(1) :
 Streptococcus pyrogens are sometimes followed by 2 important non – suppurative sequelae , acute rheumatic fever and acute
glomerulonephritis.They occur one to four weeks after acute infection.
 Strep.pyrogens is no longer detectable when complications are set in .
 Rheumatic fever is preceeded by sore throat while acute glomerulonephritis is by skin infection.
 These sequelae or complications are believed to be result of hypersensitivity to some streptococcal components.
 Rheumatic fever may follow infection with any serotype of strept.pyrogenes while acute glomerulonephritisis caused by only a
few nephritogenic types.

 ALPHA HEMOLYTIC STREPTOCOCCI :

 They produce a greenish discolouration around the colonies.,due to partial haemolysis.
 The zone of lysis is small (1 or 2 mm wide) with presence of unlysed erythrocyteswhich are detectable micro – scopically.
 Alpha haemolytic streptococci are seen in ;
 Streptococcus viridians
 Streptococcus.mitis,
 Streptococcus.anginosus,
 Streptococcus.bovis.
 Streptococcus Pneumoniae.

 NAME 2 MEDIA USED BY CULTIVATION OF CORYNEBACTERIUM DIPTHERIAE :

 These are best grown on medium enriched with blood , serum , egg.
 Growth is scanty on ordinary media
Optimum temperature - 37 c , PH = 7.2
1) Hiss serum water – liquid medium containing serum growth is seen as turbidity and pellicle formation.
2) Loffler’s serum slope - diptheria grow very rapidly.
Colonies appear after 6 – 8 hrsof incubation & colonies are small circular white creamy and glisterning.
3) Tellurite blood agar medium
Contains potassium tellurite (0.04%) inhibits other bacteria.organisms grow slowly on this medium and form grey and
black colonies due to reduction of Potassium tellurite to tellurium. Based on colony morphology on medium , 3 different
species are identified Gravis ,intermedius and mitis are distinguished.

 NAME 4 GROUPS OF E-COLI CAUSING DIARRHEAL DISEASES MORE COMMON IN INFANTS :

1) Enteropathogenic E.coli
2) Enterotoxigenic E.coli
3) Enteroinvasive E.coli
4) Enterohaemorrhagic E.coli / Verocytotoxin producing E.coli
5) Enteroaggressive E.coli
 Enterohaemorrhagic E.coli or Verocytotoxin producing E.coli (VTEC) causes haemorrhagic colitis (HC) and haemolytic uraemic
syndrome(HUS) .It is most common in infants & young children but can occur in all ages.It is characterised by marked haemorrhage
but fever is not always present.Toxins responsible are called “verotoxin” because of its effect on vero cells in culture.

 WRITE 2 IMPORTANT ORGANISMS CAUSING NOSOCOMIAL INFECTION :

 An infection acquired in a hospital or other medical facility called nasocomial infections.
 Common causative organisms for nosocomial infections are;
1) E.coli
2) Staphylococcus aureus
3) Streptococcus
4) Klebsiella
5) Pseudomonas
6) Enterobacter
7) Candida
8) Staphylococcus epidermidis
9) Bacteroids
10) Other pathogens

 ORAL LESIONS IN SYPHILIS(3) :

 Oral lesions may occur in primary , secondary , tertiary and congential syphilis.
i. Extragenital chancre of primary syphilis occurs most commonly on the lips.
ii. Secondary syphilis shows maculopapular eruptions and mucous patches in the mouth.
iii. In the tertiary syphilis , gummas or diffuse fibrosis may be seen on the hard palate and tongue.
iv. Oral lesions of the congential syphilis are fissures att the angles of mouth and characteristic peg – shaped notched
hutchinson’s incisors.

 4 SEROLOGICAL TESTS IN SYPHILIS :

1) VDRL and RPR tests are used for screening or for diagnostic purposes of large number of sera .They are also used for quantitative
measurement of reagin titre for assessment of clinical activity of syphilis.
2) Troponemal tests (TPHA or FTA – ABS ) are used to confirm the diagnosis with a positive reagin test.
a) VDRL – Veneral disease research laboratory test,
b) RPR – Rapid plasma reagin test,

15
 c) TPHA – Troponema palladium haemoagglutination assay,
d) FTA – ABS – Flourescent troponemal antibody – absorption test.

 NAME 3 GENERAL CHARACTERISTICS OF VIRUSES :

 Viruses are small obligative intracellular infective agents containing only one type of nuc leic acid as their genome.
 They have no metabolic activity outside the living cells
 They do not possess cellular organisation and lack the enzymes necessary for protein an nucleic acid synthesis.
 They multiple by a complex process not by binary fission.
 They do not grow in inanimate media ,.They are resistant to antibiotics.

 PROPHYLAXIS AGAINST HEPATITS B :

 Genral preventive measures :-
a) Screening for HBS Ag HBC Ag in blood donors.
b) Use of unsterile needles,syringes and other materials must be avoided.
 Immunisation
a) Passive immunisation:
 It is employed following any accidental exposure of HB infection.
 It may not prevent infection but protects against illness and development of carrier state.
 NOTE: HBIG is prepared from donors with high titres of anti HB’s .It can be given in doses of 300 – 500 IU.,i.m
b) Active immunisation:
 Plasma derive vaccine
 Recombinant yeast Hepatitis B vaccine(free from side effects)
 These both are immunogenic and safe.
Both vaccines are adsorbed with Al(OH)2 as adjuvant and stored in cold but not frozen.Three doses at 0,1 and 6 months are
administered i.m.,into deltoid muscle.

 FOUR INFECTIONS CAUSED BY CANDIDA(2):

1) Skin and nail infections – axillae , groin , perineum , submammary glands.
2) Nails – paronychia , onychia – seen in person who frequently immerse hands in water.
3) Systemic candidiasis – Urinary tract infections , intestinal candidiasis.
4) Oral manifestaions – Thrush , chronic oral candidiasis , chronic mucocutaneous candidiasis , angular stomatitis , circumoral
candidal dermatits.

 SYSTEMIC MYCOSIS:
 These are caused by fungi of soil and is acquired by inhalation.
 Fungus may be disseminate to CNS,bones and other internal organs.
 Systemic mycosis include blasto-mycosis, paracoccidioidomycosis, coccidioidomycosis, histoplasmosis and cryptococcosis.
 Systemic mycoses and sub cutaneous mycoses collectively are also names as ‘Deep mycoses’.

 CANDIDA ALBICANS:
 It is the causative fungus for CANDIDIASIS.
 It is an oviod (or)spherical budding yeast cell,3 – 5 µm in diameter.
 Candidiasis is an infection of skin,mucosa and internal organs,caused by yeast like fungus Candida albicans,and occasionally by
other candida species.
 It is the normal inhabitant of skin,gastrointestinal tract,oral and vaginal cavities.

 CANDIDOSIS:
 Causative organism – candida albicans (80 – 90% cases)
 It is infection of skin , mucosa and internal organs caused by yeast like fungus – candida albicans.
 It is ovoid or spherical budding yeast in diameter of 3 - 5 µm
 It is an opportunisitc endogenous infection
 Lesions caused are,
a) Mucocutaneous infections – oral thrush , vulvovaginitis.,conjuctivitis , keratitis.
b) Skin and nail infections – axillae , groin , paronychia , onychia
In infants – napkin dermatitis.
c) Systemic candidiasis – Urinary tract infections , intestinal candidiasis , meningitis , septicemia
d) Oral manifestations – Thrush , chronic oral candidiasis , Chronic mucocutaneous candidiasis.

 P.VIVAX :
 These are present in different forms in peripheral blood as Trophozoites , schizonts and gametocytes.
 Trophozoite are irregular , amoeboid , vacuole present.
 Gametocytes are spherical in shape and much larger than a red blood cell.
 Female gametocyte (Macrogametocyte) are spherical , larger than male gametocyte cytoplasm stains deep blue , nucleus is small
and compact.
 Male gametocytes (Micro gametocyte) cytoplasm stains light blue or pale blue , nucleus is large and diffuse.
 Infected erythrocytes of Plasmodium vivax are enlarged , pale , schuffner’s dots present.
 Duration of erythrocytic schizogony is 48 hours.

 TELL ABOUT 4 SPECIES OF PARASITES PRODUCING MALARIAL FEVER:

 P.vivax,
16
  P.falciparum,
 P.malariae,
 P.ovale.
 These are protozoa causing malaria in man.

 DIAGRAM OF FERTILIZED EGG OF ASCARIS LUMBRICOIDES :

 COMPLICATIONS OF PLASMODIUM FALCIPARUM :

 It is the most pathogenic of the plasmodium species infecting man.There are no relapses in p. falciparuminfection . It invades
erythrocytes of all ages(complications include pernicious anemia & Black water fever)
1) Pernicious anemia :
Life threatening complication.,it is due to heavy parasitization.Manifestations include
 Cerebral malaria : characterised by hyperpyrexia , coma , paralysis.Brain is congested.
 Algid malaria : characterised by cold clammy skin leading to peripheral circulatory failure.
2) Black water fever :
 It is manifestation of infection with P.falciparaum.occuring in those persons previously affected & has
inadequate dose of quinine.
 It is characterised by intravascular haemolysis , fever and haemoglobinuria.
 When subsequent infection infection & treatment with quinine occurs , there is massive destruction of
erythrocytes because of Antigen – Antibody reaction.

 MENTION 2 IMPORTANT MEDIA FOR CULTURING FUNGI :

 Sabouraud’s dextrose agar (SDA) and SDA medium with antibiotics are inoculated and incubated at 25 C and & 37C for 3 weeks.
 Chloramphenicol is added to supress the growth of contaminating bacteria withcycloheximide (actidione) is incorporated to supress
contaminated fungi.
 Blood heart infusion (BHI) agar with blood and antibiotics is another medium used for primary isolation of fungi.

 MENTION 4 BACTERIA CAUSING RESPIRATORY INFECTION :

 Strep.pneumoniae – causes Pneumonia in all ages,
 Strep.pyogenes – causes acute pharyngitis and Tonsilitis in all ages ,
 Corynebacterium diptheriae – causes diptheria in children,
 Klebsiella pneumoniae – causes Lobar pneumoniae in adults ,
 Bordetella pertussis – causes poroxysmal cough in children.

 WIDAL TEST :
 This test is for detection of agglutinins (H and O) in patients with enteric fever.Salmonella antibodies sharply appearing and rise at
end of first week to third week.
 Two serum specimens obtained at intervals of 7 – 10 days to read the raise of antibodies.
 Serial dilutions of unknown sera are tested against the antigens for the respective salmonella.
 Control tubes containing the antigen and normal saline are included to check for agglutination.
 The highest dilution of the serum showing agglutination (carpet formation) indicates the antibody titre against that particular
antigen.Control tubes show a compact deposit (button formation).
 Test may be positive in many healthy carriers.

 ANTI NUCLEAR ANTIBODIES(ANA) :

 Antibodies that attack healthy proteins in the nucleus (brain) of your cells are calledantinuclear antibodies (ANA).
 When the body receives signals to attack itself, autoimmune diseases such as lupus (SLE), scleroderma, mixed connective tissue
disease, and others can occur.
 An antinuclear antibodies (ANA) test is done to help identify problems with the immune system, such as: Rheumatoid arthritis.
Systemic lupus erythematosus (SLE)., Polymyositis.
 The negative test means that an active autoimmune disease is unlikely.
17
  A positive test means contains high levels of ANA in blood.

 QUICK METHOD OF DIAGNOSING CRYPTOCOCCUS NEOFORMONS:

Laboratory diagnosis specimens ;
 CSF
 Sputum
 Pus
 Brain Tissue.
 Under the microscope, the India ink stain is used for easy visualization of the capsule in cerebral spinal fluid.
 The particles of ink pigment do not enter the capsule that surrounds the spherical yeast cell, res ulting in a zone of
clearance or "halo" around the cells. This allows for quick and easy identification of C. neoformans.
 The histopathological examination of tissue can be done by staining with H & E , PAS and mucicarmine stains.

 NAME 3 GRAM POSITIVE BACILLI :

 Corynebacterium
 Clostridium
 Bacillus
 Listeriae species.

 NAME 4 DIFFERENT CLASSES OF DISINFECTANTS (1) / NAME 4 CHEMICAL DISINFECTANTS(1) :

1) Alcohols – Ethyl alcohol ,isopropyl alcohol , methyl alcohol.
2) Aldehydes – Formaldehyde , glutaraldehyde , ortho – phthalaldehyde.
3) Phenols derivatives – Cresol , chlorhexidine , chloroxylenol , hexachlorophane.
4) Halogens – Chlorine and iodine.
5) Oxidising agents – Hydrogen peroxide , peracetic acid , plasma sterilisation.
6) Salts – Salts of copper , silver , mercury .
7) Dyes – Aniline dyes , Acridine dyes.
8) Gases – Formaldehyde gas , Ethylene oxide (ETO) , Betapropiolactone(BPL).

 MENTION ALDEHYDES GROUP OF DISINFECTANTS:

 Formaldehyde ;
 It is bactericidal , sporicidal & virucidal.,used in both liquid and gaseous form.
 Uses in sterilise bacterial vaccines , prepare toxoid from toxin and preservation of tissue for histological examination.
 Glutaraldehyde ;
 It is effective against bacteria , virus , fungi , HIV , hepatitis B .
 Commercially available as ‘Cidex’ .
 Uses for sterilization of cystoscopes , endoscopes and bronchoscopes.,& plastic endotracheal tubes , face masks ,
corrugated rubber anaesthetic tubes and metal instruments.
 Ortho – phthalaldehyde ;
 High level disinfectant ., more stable during storage and rapidly mycobactericidal than glutaraldehyde.
 This is used for glutaraldehyde resistant mycobacteria.

 ZIEHL – NEELSEN STAIN:

 The acid – fast staining of mycobacteria (usually tubercle and lepra bacilli) is done for this technique.
 Reagents are : zeihl – neelsen’s carbol fuchsin , methylene blue.
 Principle – acid – fastness is due to high content of lipids , fattyacids and higher alcohols found in the cell wall of mycobacterium .
Mycolic acid (a wax) , acid – fast in the free – state , is found in all acid – fast bacteria.Besides lipid content , acid – fastness
depends also on the integrity of the cell wall.
 On microscopic examination of the smear Acid – fast bacilli appear red (colour of carbol fuchsin ) in blue(colour of methylene blue)
background of pus cells and epithelial cells.

 GRAM STAIN(1) / IMPORTANCE OF GRAMS STAIN(1) :

 Gram staining is done for identification of bacterial culture provided to you.It is the most widely used stain in bacteriology.
 Reagents are: Crystal violet solution , Gram’s iodine , Acetone , Safranin.
 There are gram positive and gram negative stains .,
Gram positive for resist decolouration and retain the colour of primary stain i.e; violet.
Gram negative are decolourised by acetone / alcohol and , therefore , take counterstain and appear red.
 Method:
 Heat fixed smear of specimen or bacterial culture is stained with crystal violet (1o stain) for one minute.Other
pararosaniline dyes such as gentian violet or methyl violet may also be used as primary stain.
 Pour gram’s iodine over the slide for one minute.
 Decolourise with acetone for 10 – 30 seconds.Alcohol can be substituted for acetone.
 Counterstain with a dye safranin for 30 seconds.Dilute carbal fuchsin or neutral red may be also used as counterstain.

 MENTION 3 IMPORTANT ACTINOMYCETIC LESIONS IN MAN :

 The actionomyces causes a disease known as actinomycosis.
 The disease occurs in four clinical forms : cervicofacial , thoracic , abdominal and pelvic actinomycosis.
 Nocardiae produce opportunistic pulmonary disease known as nocardiosis in immunocompromised individuals. It is also called
mycetoma.

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  Mycetoma is a localized chronic granulomatous disease of the subcutaneous and deeper tissues affecting commonly the foot and
presenting as a tumour with multiple discharging sinuses.

 NAME THE MICRO ORGANISMS CAUSING DENTAL CARIES:

 Many species of viridians streptococci (e.g ; Strepto.mutans) present in the oral cavity have emerged as etiological agents of dental
caries.
 Other important bacteria are Lactobacillus acidophilus and Actinomyces viscosus.
 Streptococcus mutans , Strept . sobrinus and other members of Strept . mutans causes dental plaque.
 Once the caries is intiated by Strept . mutans , the process of progression is taken up by other bacteria such as
 Lactobacilli.Strept. sanguis,
 Strept.salivarius,
 Strept.milleri,
 Strept.mitior,
 Lactobacillus casei,
 L.acidophilus,
 L.fermentum,
 Actinomyces viscosus,
 A.israelii and
 A.naeslundii are the other bacteria incriminated in the pathogenesis of dental caries.

 MENTION 2 ORAL FUNGAL INFECTIONS :

 Various manifestations of oral candidiasis are as follows:
 Oral Thrush (Pseudomembranous candidiasis),
 Chronic oral candidiasis,
 Chronic mucocutaneous candidiasis ,
 Angular stomatitis (angular chelitis) ,
 Circumoral candidal dermatits.

 MENTION THE PARASITES FOUND IN PERIPHERAL BLOOD SMEAR :

 All asexual erythrocytic stages (Ring forms , trophozoite forms , schizonts) as well as gametocytes can be found in peripheral blood
smear in infection with P.vivax ,P.malariae , and P.ovale.
 But in P.falciparum infection , only the ring forms and gametocytes (cresent shaped) can be seen.

 PERIAPICAL ABSCESS(1)&NAME MICROORGANISMS CAUSING IT :

 It is either an acute or chronic suppurative of the periapical tissues of the periodontium following pulp infection , traumatic injury of
the teeth , or irritation of the apical tissues by the mechanical or chemical manipulation.
 It is also known as Dentoalveolar abscess.
 Acute dental abscess is polymicrobial , comprising of strict anaerobes , such as anaerobic cocci , prevotella , fusobacterium
species , and facultative anaerobes , such as viridans group streptococci and streptococcus anginosus group.

 TELL ABOUT 2 IMPORTANT METHODS TRIED FOR ISOLATING MYCOBACTERIUM LEPRAE :

 The specimens from skin are obtained by slit and scrape methods.
 Skin and nerve biopsy useful In diagnosis and accurate classification of leprosy lesion.
 Animal inoculation.
 Lepromin test.
 Serological test.

 MENTION 2 ORAL FUNGAL FALCIPARUM

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