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Leishmania
Presented by:
Mahesh Yadav
M.Sc. IIYr.
Central Department of Microbiology,T.U
INTRODUCTION
• Leishmania is a genus of trypanosomatid
protozoa, which causes a fatal vector-borne parasitic
disease called Leishmaniasis .
• It is spread by the bite of sandflies of the genus
Phlebotomus in the Old World, and of the genus
Lutzomyia in the New World.
• Leishmaniasis is the second-largest parasitic killer in
the world (after malaria) and is endemic in many
parts of Africa, Asia and South America.
HISTORY
• The parasite was named by Ronald
Ross in 1903 after the Scottish
pathologist William Boog Leishman.
• In 1901, Leishman identified the
organism in smears taken from the
spleen of a patient who had died
from "dum-dum fever“.
CLASSIFICATION
• Kingdom
• Subkingdom
• Phylum
• Subphylum
• Class
• Order
• Genus
• Species
Protista
Sarcomastigophora
Protozoa
Mastigophora
zoomastigophora
Kinetplastida
Leishmania
donovani, tropica, mexicana
, braziliensis, etc.
IMPORTANT SPECIES
• L. donovani
• L. tropica
• L. mexicana
• L. braziliensis
• L.major
• L.guyanensis
• L.lainsoni
• L.naiffi
• L.aethiopica, etc
HABITAT
(L.donovani)
 Are essentially the parasites of visceral organs.
 Promastigote forms found in sand fly and in culture.
 Amastigote forms found in man in
reticuloendothelial cells of
spleen,
bone marrow,
liver,
intestinal mucosa,
mesentric lymph node.
L.donovani L. tropica L.mexicana L. braziliensis
Parasites of Visceral
organs
Skin Skin Skin and mucus
membrane of
nose and
buccal cavity
Amastigote
form found in
Human
Reticuloendothelial
cells of
•spleen,
• bone marrow ,
• liver
•intestinal
mucosa
Human
•Reticuloendothelial
cells of skin
Human
•Reticuloendothelial
cells of skin
Human
•Macrophage of skin
•Mucous membrane
of nose and buccal
cavity
Promastigote
form found in
Sand fly and
culture
Sand fly and
culture
Sand fly and
culture
Sand fly and
culture
HABITAT OF OTHER SPECIES
MORPHOLOGY
(same in all species)
• The parasite exists in
2 forms;-
1. Amastigotes –
aflagellar stage
2. Promastigotes-
flagellar stage
Morphological Differences
Amastigotes
• Aflagellar stage
• Occurs in the vertebrate host
• divides by binary fission at 37oC.
• There are round or oval ;2-4µm along longitudinal
axis.
• Nucleus relatively larger and situated centrally.
• Kinetoplast situated right angle to nucleus.
Promastigotes
• Flagellar stage
• Occurs in the sand fly
• divides by binary fission at 27oC.
• They are spindle shaped ;15-20 µm in length & 1-
2µm in width.
• Nucleus smaller and situated in the middle of
the cell or along the side of cell-wall.
• Kinetoplast lies transversely near the anterior
end.
LIFE CYCLE (L.donovani)
Life cycle of other species of Leishmania are similar to
L.donovani except that
In L.tropica
• amastigotes reside in the large mononuclear cells
of the skin
In L.mexicana
• Amastigotes found in reticuloendothelial cells
and lymphatic tissues of skin
In L.braziliensis
• amastigotes are found in reticuloendothelial cells
and lymphatic tissues of skin and mucus
membrane
MODE OFTRAMSMISSION
(L.donovani)
1. Mainly by the bite of sand fly (vector) Phlebotomus
argentipus
2. Les frequently by
• blood transfusion,
• congenital infection,
• accidental inoculation of cultured promastigotes in the lab.
workers, and
• sexual intercourse.
 Males are affected more (due to increased exposure to sand
flies through the occupation and leisure activities).
RESERVOIR
(L.donovani)
• Human:- in Indian subcontinent
• Rodents:- in Africa
• Foxes:- in Brazil and Central Asia
• Dogs :- In Mediterranean and China
L.donovani L.tropica L.mexicana L.braziliensis
Reservoir Man, rodents,
foxes, dogs
Man,
Dog
Sloth, ant
eater, rat, dog
Sloth, ant
eater, rat, dog
Vector Sand fly
Phlebotomus
argentipus
Sand fly
Phlebotomus
argentipus
Sand fly
Lutzomyia spp.,
Sand fly
Lutzomyia spp.,
Mode of
transmission
•Bite of sand
fly
•blood transfusion
•Congenital
infection
•sexual intercourse
Bite of sand fly •Bite of sand
fly,
•Bite of ticks ,
•autoinfection
•Bite of sand
fly,
•Bite of ticks ,
•autoinfection
Individual at
risk
Males are
affected more
Adolescents and
young adults
Persons working
at the edge of
forest and in the
people staying in
rural areas.
Persons working
at the edge of
forest and in the
people staying in
rural areas.
Reservoir, vector and transmission of other species
VECTOR
(Sand fly)
• Phlebotomas • Lutzomyia
CLINICAL MANIFESTATIONS
1. Pyrexia
2. Spleen enlargement
3. Lymphadenopathy
4. Darkening of the skin (KALA AZAR, MEANING “BLACK FEVER” IN HINDI, BECAUSE OF ITS
TENDENCY TO DISCOLOR ITS VICTIM’S COMPLEXION DURING ADVANCED STAGES)
5. Others:- kala-azar with HIV co-infection
Post kala-azar dermal leishmaniasis(PKDL)
 Complications:- pneumonia, TB, dysentery, uncontrolled haemorrhage
 Prognosis:- With an early treatment, cure rate >90%
If not treated, death occurs within 2 years.
CLINICAL MANIFESTATIONS OTHER SPECIES
L.tropica
• Oriental sore
• Acute necrotizing lesion
• scar
L.mexicana
• Chiclero ulcer
• Indolent nodular lesion
L.braziliensis
• Espundia
• Uta
• Pian bois
TYPES OF
LEISHMANIASIS
Leishmaniasis is divided into clinical syndromes
according to what part of the body is affected most.
Visceral Leishmaniasis(VL)
Cutaneous Leishmaniasis(CL)
Mucocutaneous leishmaniasis(MCL)
Continued....
1. Visceral Leishmaniasis (VL)
or Kala-azar
 caused by L.donovani
 part of the body affected most is
internal organs
Spleenomegaly
Continued....
2. Cutaneous
Leishmaniasis(CL)
( most common type)
a) Old world CL:- caused by
L.tropica, L. aethiopica
b) New world CL:- caused by
L.mexicana, L.braziliensis, L.g
uyanensis
c) Dermal leishmanoid or Post
kala-azar dermal
leishmaniasis(PKDL):- caused
by L.donovani
 Part of the body most affected
is skin
....continued
3. Mucocutaneous
leishmaniasis(MCL)
 Caused by L. braziliensis
and occasionally by
L.panamensis
 Part of the body affected
most is skin and mucous
membrane of nose and
pharynx
SYNONYMS OF LEISHMANIASIS
Cutaneous
leishmaniasis
Aleppo boil,
Baghdad boil,
Delhi boil,
Kandahar sore,
Lahore sore,
Oriental sore,
Visceral
leishmaniasis
Kala-azar,
Black fever
Dum-Dum fever,
Sahib’s disease
Kala Dukh
White leprosy
Mucocutaneous
Leishmaniasis
Breda's disease
bosch yaws,
bush yaws
forest yaws
LABORATORY DIAGNOSIS
Direct Evidences
Peripheral blood by thick
film
method.(Amastigote
form)
Blood culture in N.N.N.
Medium. (Promastigote
form)
Biopsy material obtained by
lymph node
puncture, sternal or iliac
crest puncture(marrow)
and spleen puncture(spleen
pulp) only for L. donovani
Indirect
evidences
Blood count
Serum Tests
Other
methods
Animal
inoculation
Leishmanin or
Montenegro
Test
Adler’s test
Direct Evidences (contd......)
1. Peripheral blood by thick
film method.(Amastigote
form)
Amastigotes in a macrophage
Direct Evidences (contd......)
2. Blood culture in N.N.N.
Medium. (Promastigote
form)
Promastigote from culture in NNN medium
Direct Evidences (....contd)
3. Biopsy material obtained
by
• lymph node puncture,
• sternal or iliac crest
puncture(marrow) and
• spleen puncture(spleen
pulp)
Amastigote form in a stained smear
Promastigote in culture in NNN medium
Amastigotes of L. donovani.
Splenic aspirate.
Indirect evidences
1. Blood count:-
• Leucopenia (progressive)
• Anaemia (raised ESR)
2. Serum Tests
• Aldehyde test- positive after 3
months.
• Antimony test- less reliable. Not
used now.
• Complement fixation test with
W.K.K. antigen. Not used now.
• Demonstration of antibodies
(ELISA, DAT, IHA, IFA with specific
antigen etc.)
• Molecular diagnosis:- DNA
Probes, PCR, etc.
Other methods
• Animal inoculation Wherever in vitro facilities
are not there, the material from patients can be injected
intraperitoneally in hamster or mice and the parasite is recovered
from the animal. In positive cases, the amastigotes can be
demonstrated in the stained impression smears of spleen from
animals.
• Leishmaninor Montenegro Test
It is a delayed hypersensitivity test. 0.2 ml of leishmania antigen
is injected intradermally. The test is read after 48-72 hrs. Positive
result is indicated by an induration of 5mm or more. In kala-azar
(visceral leishmaniasis), this test is negative
• Adler’s test:- It is a serological method. The
development of promastigote forms of Leishmania in Locke’s serum
agar can be inhibited by a immune serum specific to
L.donovani, L.tropica and L.braziliensis.
EPIDEMIOLOGY
• Found in more than 88 countries.
• Found on every continent except Australia and Antarctica.
• For cutaneous leishmaniasis, number of cases range from 0.7 million to 1.2 million .
• For visceral leishmaniasis, number of cases range from 0.2 million to 0.4 million.
• Annual incidence of disease= 600,000 cases per year.
• People infected worldwide=12 million.
• People at risk=350 million.
GEOGRAPHICAL DISTRIBUTIONS
• Present worldwide.
• Most of the affected countries are in the tropics and subtropics.
• More than 90 percent of the world's cases of visceral leishmaniasis are in
India, Bangladesh, Nepal, Sudan, and Brazil.
Leishmaniasis is found in
• Asia (not Southeast Asia),
• Central America,
• South America (not in Uruguay, Chile, or Canada),
• Southern Europe (not common in travelers to southern Europe),
• The Middle East, and
• Africa (particularly East and North Africa, )
Current Geographic Distribution of
Leishmaniasis (....contd)
Leishmaniasis in Nepal
• Visceral leishmaniasis (kala azar) is common in the Terai region.
• The first confirmed case of VL was recorded in 1980.
• A total of 25890 cases with 599 deaths were reported during 1980-2006.
• During 2003, highest incidence (per 100,000) was in Mahottari district (184), followed by
Sarlahi (100) and Sunsari (96).
• Highest case fatality rate (CFR) was in Dhanusha (2.9%) followed by Bara (2.4%) and Saptari
(2.0%).
• Cutaneous leishmaniasis is rare in Nepal.
• First case of cutaneous leishmaniasis was reported in the year 2006 in Nepal.
Leishmaniasis in Nepal
(...contd.)
Reduction of sand
fly population
by insecticides mainly DDT,
dieldrin, malathion
Reduction of
reservoir
by killing all the infected dogs in the
cases of zoonotic kala-azar.
Education in the
community
About the causes and modes of
transmission of leishmaniasis.
Prevention of
exposure to sand fly
using insect repellent, bed nets and window
mess as needed.
PREVENTION AND
CONTROL
PREVENTION AND CONTROL
There are No Vaccines to prevent leishmaniasis.
PREVENTION AND CONTROL
(.....contd.)
TREATMENT
Drugs
Sodium
stibogluconate
solution
Inhibits glycolytic enzymes and fatty acid
oxidation
Amphotericin
B
Binds with ergosterol leading to the altered
permeability to cations, water, glucose and affect
membrane-bound enzymes.
Pentamidine Inhibits DHFR and interferes with aerobic glycolysis
in protozoa, also inhibits protein synthesis
Miltefosine Effects cell-signaling pathways and synthesis of the
cell-membrane
Interferon
macrophage activation
Specific therapy
supplemented with
treatment of secondary
microbial infections
high-calorie-high protein
diet
Blood transfusion in
severe anaemia
TREATMENT (....contd)
REFERENCES
• www.who.int
• www.cdc.gov
• www.Leishinfonet.com
• Chatterjee KD,(2009), Parasitology protozoology and
helminthology, 13th edition, CBS publishers and
distributers pvt. Ltd. New Delhi, India Page no.64-89
• Parija S.C., (2004), Textbook of Medical Parasitology, 2nd
edition, All India publishers and Distributers. Page No.81-
103
THANK YOU

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Leishmania presented by Mahesh

  • 1. Leishmania Presented by: Mahesh Yadav M.Sc. IIYr. Central Department of Microbiology,T.U
  • 2. INTRODUCTION • Leishmania is a genus of trypanosomatid protozoa, which causes a fatal vector-borne parasitic disease called Leishmaniasis . • It is spread by the bite of sandflies of the genus Phlebotomus in the Old World, and of the genus Lutzomyia in the New World. • Leishmaniasis is the second-largest parasitic killer in the world (after malaria) and is endemic in many parts of Africa, Asia and South America.
  • 3. HISTORY • The parasite was named by Ronald Ross in 1903 after the Scottish pathologist William Boog Leishman. • In 1901, Leishman identified the organism in smears taken from the spleen of a patient who had died from "dum-dum fever“.
  • 4. CLASSIFICATION • Kingdom • Subkingdom • Phylum • Subphylum • Class • Order • Genus • Species Protista Sarcomastigophora Protozoa Mastigophora zoomastigophora Kinetplastida Leishmania donovani, tropica, mexicana , braziliensis, etc.
  • 5. IMPORTANT SPECIES • L. donovani • L. tropica • L. mexicana • L. braziliensis • L.major • L.guyanensis • L.lainsoni • L.naiffi • L.aethiopica, etc
  • 6. HABITAT (L.donovani)  Are essentially the parasites of visceral organs.  Promastigote forms found in sand fly and in culture.  Amastigote forms found in man in reticuloendothelial cells of spleen, bone marrow, liver, intestinal mucosa, mesentric lymph node.
  • 7. L.donovani L. tropica L.mexicana L. braziliensis Parasites of Visceral organs Skin Skin Skin and mucus membrane of nose and buccal cavity Amastigote form found in Human Reticuloendothelial cells of •spleen, • bone marrow , • liver •intestinal mucosa Human •Reticuloendothelial cells of skin Human •Reticuloendothelial cells of skin Human •Macrophage of skin •Mucous membrane of nose and buccal cavity Promastigote form found in Sand fly and culture Sand fly and culture Sand fly and culture Sand fly and culture HABITAT OF OTHER SPECIES
  • 8. MORPHOLOGY (same in all species) • The parasite exists in 2 forms;- 1. Amastigotes – aflagellar stage 2. Promastigotes- flagellar stage
  • 9. Morphological Differences Amastigotes • Aflagellar stage • Occurs in the vertebrate host • divides by binary fission at 37oC. • There are round or oval ;2-4µm along longitudinal axis. • Nucleus relatively larger and situated centrally. • Kinetoplast situated right angle to nucleus. Promastigotes • Flagellar stage • Occurs in the sand fly • divides by binary fission at 27oC. • They are spindle shaped ;15-20 µm in length & 1- 2µm in width. • Nucleus smaller and situated in the middle of the cell or along the side of cell-wall. • Kinetoplast lies transversely near the anterior end.
  • 11. Life cycle of other species of Leishmania are similar to L.donovani except that In L.tropica • amastigotes reside in the large mononuclear cells of the skin In L.mexicana • Amastigotes found in reticuloendothelial cells and lymphatic tissues of skin In L.braziliensis • amastigotes are found in reticuloendothelial cells and lymphatic tissues of skin and mucus membrane
  • 12. MODE OFTRAMSMISSION (L.donovani) 1. Mainly by the bite of sand fly (vector) Phlebotomus argentipus 2. Les frequently by • blood transfusion, • congenital infection, • accidental inoculation of cultured promastigotes in the lab. workers, and • sexual intercourse.  Males are affected more (due to increased exposure to sand flies through the occupation and leisure activities).
  • 13. RESERVOIR (L.donovani) • Human:- in Indian subcontinent • Rodents:- in Africa • Foxes:- in Brazil and Central Asia • Dogs :- In Mediterranean and China
  • 14. L.donovani L.tropica L.mexicana L.braziliensis Reservoir Man, rodents, foxes, dogs Man, Dog Sloth, ant eater, rat, dog Sloth, ant eater, rat, dog Vector Sand fly Phlebotomus argentipus Sand fly Phlebotomus argentipus Sand fly Lutzomyia spp., Sand fly Lutzomyia spp., Mode of transmission •Bite of sand fly •blood transfusion •Congenital infection •sexual intercourse Bite of sand fly •Bite of sand fly, •Bite of ticks , •autoinfection •Bite of sand fly, •Bite of ticks , •autoinfection Individual at risk Males are affected more Adolescents and young adults Persons working at the edge of forest and in the people staying in rural areas. Persons working at the edge of forest and in the people staying in rural areas. Reservoir, vector and transmission of other species
  • 16. CLINICAL MANIFESTATIONS 1. Pyrexia 2. Spleen enlargement 3. Lymphadenopathy 4. Darkening of the skin (KALA AZAR, MEANING “BLACK FEVER” IN HINDI, BECAUSE OF ITS TENDENCY TO DISCOLOR ITS VICTIM’S COMPLEXION DURING ADVANCED STAGES) 5. Others:- kala-azar with HIV co-infection Post kala-azar dermal leishmaniasis(PKDL)  Complications:- pneumonia, TB, dysentery, uncontrolled haemorrhage  Prognosis:- With an early treatment, cure rate >90% If not treated, death occurs within 2 years.
  • 17. CLINICAL MANIFESTATIONS OTHER SPECIES L.tropica • Oriental sore • Acute necrotizing lesion • scar L.mexicana • Chiclero ulcer • Indolent nodular lesion L.braziliensis • Espundia • Uta • Pian bois
  • 18. TYPES OF LEISHMANIASIS Leishmaniasis is divided into clinical syndromes according to what part of the body is affected most. Visceral Leishmaniasis(VL) Cutaneous Leishmaniasis(CL) Mucocutaneous leishmaniasis(MCL)
  • 19. Continued.... 1. Visceral Leishmaniasis (VL) or Kala-azar  caused by L.donovani  part of the body affected most is internal organs Spleenomegaly
  • 20. Continued.... 2. Cutaneous Leishmaniasis(CL) ( most common type) a) Old world CL:- caused by L.tropica, L. aethiopica b) New world CL:- caused by L.mexicana, L.braziliensis, L.g uyanensis c) Dermal leishmanoid or Post kala-azar dermal leishmaniasis(PKDL):- caused by L.donovani  Part of the body most affected is skin
  • 21. ....continued 3. Mucocutaneous leishmaniasis(MCL)  Caused by L. braziliensis and occasionally by L.panamensis  Part of the body affected most is skin and mucous membrane of nose and pharynx
  • 22. SYNONYMS OF LEISHMANIASIS Cutaneous leishmaniasis Aleppo boil, Baghdad boil, Delhi boil, Kandahar sore, Lahore sore, Oriental sore, Visceral leishmaniasis Kala-azar, Black fever Dum-Dum fever, Sahib’s disease Kala Dukh White leprosy Mucocutaneous Leishmaniasis Breda's disease bosch yaws, bush yaws forest yaws
  • 23. LABORATORY DIAGNOSIS Direct Evidences Peripheral blood by thick film method.(Amastigote form) Blood culture in N.N.N. Medium. (Promastigote form) Biopsy material obtained by lymph node puncture, sternal or iliac crest puncture(marrow) and spleen puncture(spleen pulp) only for L. donovani Indirect evidences Blood count Serum Tests Other methods Animal inoculation Leishmanin or Montenegro Test Adler’s test
  • 24. Direct Evidences (contd......) 1. Peripheral blood by thick film method.(Amastigote form) Amastigotes in a macrophage
  • 25. Direct Evidences (contd......) 2. Blood culture in N.N.N. Medium. (Promastigote form) Promastigote from culture in NNN medium
  • 26. Direct Evidences (....contd) 3. Biopsy material obtained by • lymph node puncture, • sternal or iliac crest puncture(marrow) and • spleen puncture(spleen pulp) Amastigote form in a stained smear Promastigote in culture in NNN medium Amastigotes of L. donovani. Splenic aspirate.
  • 27. Indirect evidences 1. Blood count:- • Leucopenia (progressive) • Anaemia (raised ESR) 2. Serum Tests • Aldehyde test- positive after 3 months. • Antimony test- less reliable. Not used now. • Complement fixation test with W.K.K. antigen. Not used now. • Demonstration of antibodies (ELISA, DAT, IHA, IFA with specific antigen etc.) • Molecular diagnosis:- DNA Probes, PCR, etc.
  • 28. Other methods • Animal inoculation Wherever in vitro facilities are not there, the material from patients can be injected intraperitoneally in hamster or mice and the parasite is recovered from the animal. In positive cases, the amastigotes can be demonstrated in the stained impression smears of spleen from animals. • Leishmaninor Montenegro Test It is a delayed hypersensitivity test. 0.2 ml of leishmania antigen is injected intradermally. The test is read after 48-72 hrs. Positive result is indicated by an induration of 5mm or more. In kala-azar (visceral leishmaniasis), this test is negative • Adler’s test:- It is a serological method. The development of promastigote forms of Leishmania in Locke’s serum agar can be inhibited by a immune serum specific to L.donovani, L.tropica and L.braziliensis.
  • 29. EPIDEMIOLOGY • Found in more than 88 countries. • Found on every continent except Australia and Antarctica. • For cutaneous leishmaniasis, number of cases range from 0.7 million to 1.2 million . • For visceral leishmaniasis, number of cases range from 0.2 million to 0.4 million. • Annual incidence of disease= 600,000 cases per year. • People infected worldwide=12 million. • People at risk=350 million.
  • 30. GEOGRAPHICAL DISTRIBUTIONS • Present worldwide. • Most of the affected countries are in the tropics and subtropics. • More than 90 percent of the world's cases of visceral leishmaniasis are in India, Bangladesh, Nepal, Sudan, and Brazil. Leishmaniasis is found in • Asia (not Southeast Asia), • Central America, • South America (not in Uruguay, Chile, or Canada), • Southern Europe (not common in travelers to southern Europe), • The Middle East, and • Africa (particularly East and North Africa, )
  • 31. Current Geographic Distribution of Leishmaniasis (....contd)
  • 32. Leishmaniasis in Nepal • Visceral leishmaniasis (kala azar) is common in the Terai region. • The first confirmed case of VL was recorded in 1980. • A total of 25890 cases with 599 deaths were reported during 1980-2006. • During 2003, highest incidence (per 100,000) was in Mahottari district (184), followed by Sarlahi (100) and Sunsari (96). • Highest case fatality rate (CFR) was in Dhanusha (2.9%) followed by Bara (2.4%) and Saptari (2.0%). • Cutaneous leishmaniasis is rare in Nepal. • First case of cutaneous leishmaniasis was reported in the year 2006 in Nepal.
  • 34. Reduction of sand fly population by insecticides mainly DDT, dieldrin, malathion Reduction of reservoir by killing all the infected dogs in the cases of zoonotic kala-azar. Education in the community About the causes and modes of transmission of leishmaniasis. Prevention of exposure to sand fly using insect repellent, bed nets and window mess as needed. PREVENTION AND CONTROL PREVENTION AND CONTROL There are No Vaccines to prevent leishmaniasis.
  • 36. TREATMENT Drugs Sodium stibogluconate solution Inhibits glycolytic enzymes and fatty acid oxidation Amphotericin B Binds with ergosterol leading to the altered permeability to cations, water, glucose and affect membrane-bound enzymes. Pentamidine Inhibits DHFR and interferes with aerobic glycolysis in protozoa, also inhibits protein synthesis Miltefosine Effects cell-signaling pathways and synthesis of the cell-membrane Interferon macrophage activation
  • 37. Specific therapy supplemented with treatment of secondary microbial infections high-calorie-high protein diet Blood transfusion in severe anaemia TREATMENT (....contd)
  • 38. REFERENCES • www.who.int • www.cdc.gov • www.Leishinfonet.com • Chatterjee KD,(2009), Parasitology protozoology and helminthology, 13th edition, CBS publishers and distributers pvt. Ltd. New Delhi, India Page no.64-89 • Parija S.C., (2004), Textbook of Medical Parasitology, 2nd edition, All India publishers and Distributers. Page No.81- 103