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PLASMODIUM
 An ancient disease : substantial toll of human life and
sufferings
 Originated from Italian word Mala (bad) and ayia(air)
 Latin word : Marshy
 So
 , malaria : disease caused by heat humidity and marshy areas
 1st discovered : Alphonse Laveran : 1880 in RBC of patient
(Algeria)
 Transmitted by female Anopheles mosquito
 Common insect borne infection
 Most deadly vector borne disease in the world
 Life threatening parasitic problem : global problem worldwide
 40% of world’s population : (2.4) billion risk
 400-900 million people are affected
 Kingdom: Animalia
 Phylum: Apicomplexa
 Class: Coccidia
 Order: Haemosporidia
 Genus: Plasmodium
 Species: vivax, falciparum, ovale , malariae
 P.vivax and P.falciparum : account 95% of infection
 Some estimate : P.vivax : accounts 80% of infections : widely
distributed in tropics, subtropics and temperate zones
Different stages:
1.Pre-Erythryocytic Schizogony
2. Erythrocytic Schizogony
3. Gametogony
4. Exo-Erythryocytic Schizogony (P.vivax, P.ovale)
1.Pre-erythrocytic Schizogony
 1st stage of human cycle
 Sporozoites : doesn’t directly enter into RBC : so k/as PES
occurs : inside parenchyma of cells
 Fully developed schizont measures 42µm : contains large no of
merozoites
 Smaller micromerozoites : enter into circulation : to start ES
 Larger macromerozoites : Re-enter liver cells : to start Exo ES
 Some sporozoites : remain dormant in liver : Hypnozoites : cause relapse
Duration of this phase :
 P.falciparum : 6 days
 P.vivax : 8 days
 P.ovale : 9days
 P. malariae :13-16days
2. Erythrocytic schizogony
 Infected liver ruptures : merozoites release : invade RBC’s
 Parasite reside the RBC and passes through :
RBC Trophozoite Schizont Merozoite
 P.vivax : greater tendency for younger erythrocytes and
reticulocytes
 P.falciparum : any age
 P.malariae : old
 P.ovale : Young
 Parasitised red cells : enlarged : cells mature with parasites :
show stippling(formation of small dot)
 P.vivax : Schuffner’s dot
 P.falciparum : Maurer’s dot (large red spots)
 P.malariae : Ziemann’s dot (few tiny dots)
 P.ovale : Schuffner’s dot
 Trophozoite : Have active amoeboid
 2 forms :
1. Ring form (early trophozoite)
 Nucleus : thinner side of ring
2. Amoeboid form (late trophozoite)
 Presence of pseudopodia
 Contains malarial pigment
Schizont
 Appears after a period of 36-40 hours
 Full grown trophozoite : ready to divide
 Round in shape
 Lost all amoeboid activities
 Nucleus is large and lie at periphery
 2 form :
Immature schizont (Nucleus not divided)
Mature schizont : Nucleus divided
3.Gametogony
 After ES : some merozoites : give rise to gametocytes : sexual function
after leaving man host
 Occurs inside capillaries of bone marrow and spleen
 Mature gametocytes : appears in peripheral blood
 Microgametocyte (male) : boarder, shorter with blunt ends
 Macrogametocyte (female): longer, narrower, pointed ends
 Changes in infected RBC’s (increase in size, pallor and different dots )
4.Exo-erythrocytic Schizogony
 Resembles PE form in morphology
 Maintained upto 3 yrs and independent of ES
 Short term and long term relapses (deteriorate after a period of
improvement)
Sporozoite PES Development of hypnozoites
ES EES
primary malaria Relapses
Relapse :
 in case of P.vivax and P.ovale
due to the presence of hypnozoites
Recrudescence :
 situation : RBC infection is not eliminated by the immune
system or by therapy
 No of RBCs begin to increase again with subsequent clinical
symptoms
 All species may cause
 Sexual cycle of malarial parasite
 Starts in human body : formation of gametocytes
 Mosquito : blood meal : ingests both sexual and asexual forms
 Asexual forms : digested
 Sexual forms (gametocyte) : undergo further development
 Blood of human carrier : must contain 12 gametocytes/mm3
 No of female gametocytes more than male gametocytes
 1st phase : mid-gut of stomach
 Nucleus of each male gametocyte : 8 long flagellates(microgamete)
: highly motile
 Process : observed : outside mosquito : thick film: exflagellation
 Female gametocyte : don't divide : Macrogamete
 Fertilizes : Zygote : motion less: later becomes motile : Ookinite
 Ookinite : migrates to stomach wall : oocyst
 Large no of sporozoites inside oocyst
 When fully mature : oocyst ruptures : liberates sporozoites :
spread all parts : salivary gland
 Ready to be transmitted : when it takes blood meal
MOT : bite of Anopheles mosquito
Extrinsic Incubation period :
 different periods for the development of sexual cycle at given
temp
 Varies : 8 to 21 days
Incubation period
 P. falciparum : 12 days(9-14 days)
 P. vivax : 14 days (8-17 days)
 P. malariae : 28 days (18-40 days)
 P. ovale :17 days (16-18days)
Main features : fever peaks followed by anemia and splenomegaly
Mild to severe and complicated :
 According to species of parasite present
 Patient’s state of immunity
 Certain disease like : malnutrition and other disease
 Severe in children and pregnancy
Main clinical features
1.Prodromal period
 Malarial paroxysm :preceed by prodromal period
 Non-specific symptoms : malaise, myalgia, headache and
fatigue
 Some localized symptoms :chest pain, abdominal pain and
arthalgia
2. Malarial paroxysm
 Classical manifestation of acute malaria
 Characterised by fever chills and rigors
Primary fever
 Typical attack 3 distinct stages: cold stage, hot stage and
sweating stage
a. Cold stage :
 Onset with lassitude (lethargy), headache, nausea and chilly
sensations followed in an hour or so by rigors
b. Hot stage :
 Patient feels hot and the skin is hot and dry to touch
 Headache intense
 Lasts for 30 min to 6 hrs
c. Sweating stage
 Profuse sweating follows the hot stage
 Continues for hour or so
 Temp drops rapidly to normal
 Skin is cool and moist
So, primary attack follows a febrile interval of 48-72hrs
3.Anemia
 Normocytic normochromic anemia
 Severe in falciparum malaria
4. Hepatospleenomegaly
 Spleen : palpable after 2nd weeks of fever
 Severe in P.falciparum : so K/as malignat malaria
5. Malaria in pregnancy
 Miscarriage or abortion
6.Malaria in children
 More severe than as in adults
 May develop convulsion (muscular contarction) during malarial
attack
 Dehydration: as a result of vomiting and sweating.
 P.vivax : Benign tertiary malaria : 48hrs
 P.falciparum : Malignant tertiary malaria:36-48hrs
 P.malariae : Benign Quartan Malaria : 72 hours
 P.ovale : Benign Tertian Malaria : 48 hrs
1.Black water fever
 Repeated infection of P.falciparum: inadequately treated
with quinine
 Massive hemolysis followed by fever and
haemoglobinuria(black coloured urine),hyperbilirubenemia
 Complication : uraemia (blood poisoning), renal failure
,anemia, pigment calculi
2.Pernicious anemia (Cerebral malaria or algid malaria)
May be different forms :
a. Pernicious malaria affecting nervous system : cerebral malaria
b. Pernicious malaria affecting GIT system (algid malaria)
c. Pernicious types affecting cardiovascular, respiratory and
genitourinary tract
Specimen :
 Blood (before antimalarial drug)
 Earlobe or finger in adults
 Toe in infants
 Collected : peak fever
 More imp : frequently examination of blood smear
1.Light microscopy
2. Fluorescence microscopy
3. Quantitative buffy coat
1.Light microscope
 Blood smear
 Gold standard method
 Most commonly used
 Depends upon : demonstration of parasite in stained PBS
 Ring forms and gametocyte : commonly seen in PBS
1.Thick smear
 Smear preparation
 Dehaemoglobinisation with d/w
 Dried and stained with Romanowsky’s stain : Leishman stain.
Geimsa stain
Uses
 To detect parasite
 Demonstrating malarial pigment
P. falciparum : (only ring and crescent form)
 Many ring forms
 Crescent forms gametocyte
 Malarial pigments : inside the blood
P. vivax
 Trophozoites, Schizont and Gametocytes can be seen in PBS
 Ring form : nucleus more thicker
 Gametocyte : spherical or globular
 Schufnner’s dot
Thin smear
 Rapidly dried
 Fixed in alcohol and stained
Uses
 Detecting parasites
 Identify species
P. falciparum
 Ring form alone or along with gametocytes
 Multiple rings in individual RBC’s
 Presence of Maurer’s dot
 Banana shaped gametocytes
The rings are small:
1/3 or less of the rbc
Ameboid trophozoite, enlarged RBC
, Schüffner’s dots
Gametocyte
More than 20 merozoites
2. Fluorescence microscope
 Kawamoto technique : fluorescent staining methods : malarial
parasite
 Blood smear : slide : stain with Acridine orange
 Nuclear DNA : green
 Cytoplasmic RNA : Red
 Examined under fluorescence microscope
3. Quantitative buffy coat (QBC)method
 Sensitive method
 Based on : ability of acridine orange to stain
nucleic acid containing parasites
 Blood : capillary tube : coated with fluorescent
dye : microhaematocrit centrifugation
 Buffy coat : observed under fluorescent
microscope
 Acridine orange stained malarial parasite :
brilliant green
4.Serodiagnosis
 IHA
 ELISA
 IFA
5. Rapid diagnostic kit
6. Dipstick method
 An enzyme immunoassay : which detects histidine rich protein 2
Pf(HRP-2) : metabolic product produced by P.falciparum
7. Molecular diagnosis
 DNA and RNA : PCR
 Chloroquine
 Amodiquine
 Proguanil
 Quinine
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Ancient Parasitic Disease: The Global Burden of Malaria

  • 2.  An ancient disease : substantial toll of human life and sufferings  Originated from Italian word Mala (bad) and ayia(air)  Latin word : Marshy  So  , malaria : disease caused by heat humidity and marshy areas  1st discovered : Alphonse Laveran : 1880 in RBC of patient (Algeria)
  • 3.  Transmitted by female Anopheles mosquito  Common insect borne infection  Most deadly vector borne disease in the world  Life threatening parasitic problem : global problem worldwide  40% of world’s population : (2.4) billion risk  400-900 million people are affected
  • 4.  Kingdom: Animalia  Phylum: Apicomplexa  Class: Coccidia  Order: Haemosporidia  Genus: Plasmodium  Species: vivax, falciparum, ovale , malariae
  • 5.
  • 6.  P.vivax and P.falciparum : account 95% of infection  Some estimate : P.vivax : accounts 80% of infections : widely distributed in tropics, subtropics and temperate zones
  • 7.
  • 8.
  • 9.
  • 10. Different stages: 1.Pre-Erythryocytic Schizogony 2. Erythrocytic Schizogony 3. Gametogony 4. Exo-Erythryocytic Schizogony (P.vivax, P.ovale)
  • 11. 1.Pre-erythrocytic Schizogony  1st stage of human cycle  Sporozoites : doesn’t directly enter into RBC : so k/as PES occurs : inside parenchyma of cells  Fully developed schizont measures 42µm : contains large no of merozoites  Smaller micromerozoites : enter into circulation : to start ES  Larger macromerozoites : Re-enter liver cells : to start Exo ES  Some sporozoites : remain dormant in liver : Hypnozoites : cause relapse
  • 12. Duration of this phase :  P.falciparum : 6 days  P.vivax : 8 days  P.ovale : 9days  P. malariae :13-16days
  • 13. 2. Erythrocytic schizogony  Infected liver ruptures : merozoites release : invade RBC’s  Parasite reside the RBC and passes through : RBC Trophozoite Schizont Merozoite  P.vivax : greater tendency for younger erythrocytes and reticulocytes  P.falciparum : any age  P.malariae : old  P.ovale : Young
  • 14.  Parasitised red cells : enlarged : cells mature with parasites : show stippling(formation of small dot)  P.vivax : Schuffner’s dot  P.falciparum : Maurer’s dot (large red spots)  P.malariae : Ziemann’s dot (few tiny dots)  P.ovale : Schuffner’s dot
  • 15.  Trophozoite : Have active amoeboid  2 forms : 1. Ring form (early trophozoite)  Nucleus : thinner side of ring 2. Amoeboid form (late trophozoite)  Presence of pseudopodia  Contains malarial pigment
  • 16. Schizont  Appears after a period of 36-40 hours  Full grown trophozoite : ready to divide  Round in shape  Lost all amoeboid activities  Nucleus is large and lie at periphery  2 form : Immature schizont (Nucleus not divided) Mature schizont : Nucleus divided
  • 17. 3.Gametogony  After ES : some merozoites : give rise to gametocytes : sexual function after leaving man host  Occurs inside capillaries of bone marrow and spleen  Mature gametocytes : appears in peripheral blood  Microgametocyte (male) : boarder, shorter with blunt ends  Macrogametocyte (female): longer, narrower, pointed ends  Changes in infected RBC’s (increase in size, pallor and different dots )
  • 18. 4.Exo-erythrocytic Schizogony  Resembles PE form in morphology  Maintained upto 3 yrs and independent of ES  Short term and long term relapses (deteriorate after a period of improvement) Sporozoite PES Development of hypnozoites ES EES primary malaria Relapses
  • 19. Relapse :  in case of P.vivax and P.ovale due to the presence of hypnozoites Recrudescence :  situation : RBC infection is not eliminated by the immune system or by therapy  No of RBCs begin to increase again with subsequent clinical symptoms  All species may cause
  • 20.  Sexual cycle of malarial parasite  Starts in human body : formation of gametocytes  Mosquito : blood meal : ingests both sexual and asexual forms  Asexual forms : digested  Sexual forms (gametocyte) : undergo further development  Blood of human carrier : must contain 12 gametocytes/mm3
  • 21.  No of female gametocytes more than male gametocytes  1st phase : mid-gut of stomach  Nucleus of each male gametocyte : 8 long flagellates(microgamete) : highly motile  Process : observed : outside mosquito : thick film: exflagellation  Female gametocyte : don't divide : Macrogamete  Fertilizes : Zygote : motion less: later becomes motile : Ookinite
  • 22.  Ookinite : migrates to stomach wall : oocyst  Large no of sporozoites inside oocyst  When fully mature : oocyst ruptures : liberates sporozoites : spread all parts : salivary gland  Ready to be transmitted : when it takes blood meal
  • 23. MOT : bite of Anopheles mosquito Extrinsic Incubation period :  different periods for the development of sexual cycle at given temp  Varies : 8 to 21 days Incubation period  P. falciparum : 12 days(9-14 days)  P. vivax : 14 days (8-17 days)  P. malariae : 28 days (18-40 days)  P. ovale :17 days (16-18days)
  • 24. Main features : fever peaks followed by anemia and splenomegaly Mild to severe and complicated :  According to species of parasite present  Patient’s state of immunity  Certain disease like : malnutrition and other disease  Severe in children and pregnancy
  • 25. Main clinical features 1.Prodromal period  Malarial paroxysm :preceed by prodromal period  Non-specific symptoms : malaise, myalgia, headache and fatigue  Some localized symptoms :chest pain, abdominal pain and arthalgia 2. Malarial paroxysm  Classical manifestation of acute malaria  Characterised by fever chills and rigors
  • 26. Primary fever  Typical attack 3 distinct stages: cold stage, hot stage and sweating stage a. Cold stage :  Onset with lassitude (lethargy), headache, nausea and chilly sensations followed in an hour or so by rigors b. Hot stage :  Patient feels hot and the skin is hot and dry to touch  Headache intense  Lasts for 30 min to 6 hrs
  • 27. c. Sweating stage  Profuse sweating follows the hot stage  Continues for hour or so  Temp drops rapidly to normal  Skin is cool and moist So, primary attack follows a febrile interval of 48-72hrs
  • 28. 3.Anemia  Normocytic normochromic anemia  Severe in falciparum malaria 4. Hepatospleenomegaly  Spleen : palpable after 2nd weeks of fever  Severe in P.falciparum : so K/as malignat malaria 5. Malaria in pregnancy  Miscarriage or abortion 6.Malaria in children  More severe than as in adults  May develop convulsion (muscular contarction) during malarial attack  Dehydration: as a result of vomiting and sweating.
  • 29.  P.vivax : Benign tertiary malaria : 48hrs  P.falciparum : Malignant tertiary malaria:36-48hrs  P.malariae : Benign Quartan Malaria : 72 hours  P.ovale : Benign Tertian Malaria : 48 hrs
  • 30. 1.Black water fever  Repeated infection of P.falciparum: inadequately treated with quinine  Massive hemolysis followed by fever and haemoglobinuria(black coloured urine),hyperbilirubenemia  Complication : uraemia (blood poisoning), renal failure ,anemia, pigment calculi
  • 31. 2.Pernicious anemia (Cerebral malaria or algid malaria) May be different forms : a. Pernicious malaria affecting nervous system : cerebral malaria b. Pernicious malaria affecting GIT system (algid malaria) c. Pernicious types affecting cardiovascular, respiratory and genitourinary tract
  • 32. Specimen :  Blood (before antimalarial drug)  Earlobe or finger in adults  Toe in infants  Collected : peak fever  More imp : frequently examination of blood smear
  • 33. 1.Light microscopy 2. Fluorescence microscopy 3. Quantitative buffy coat 1.Light microscope  Blood smear  Gold standard method  Most commonly used  Depends upon : demonstration of parasite in stained PBS  Ring forms and gametocyte : commonly seen in PBS
  • 34. 1.Thick smear  Smear preparation  Dehaemoglobinisation with d/w  Dried and stained with Romanowsky’s stain : Leishman stain. Geimsa stain Uses  To detect parasite  Demonstrating malarial pigment
  • 35.
  • 36. P. falciparum : (only ring and crescent form)  Many ring forms  Crescent forms gametocyte  Malarial pigments : inside the blood P. vivax  Trophozoites, Schizont and Gametocytes can be seen in PBS  Ring form : nucleus more thicker  Gametocyte : spherical or globular  Schufnner’s dot
  • 37.
  • 38.
  • 39.
  • 40.
  • 41. Thin smear  Rapidly dried  Fixed in alcohol and stained Uses  Detecting parasites  Identify species P. falciparum  Ring form alone or along with gametocytes  Multiple rings in individual RBC’s  Presence of Maurer’s dot  Banana shaped gametocytes
  • 42.
  • 43.
  • 44. The rings are small: 1/3 or less of the rbc
  • 45.
  • 46. Ameboid trophozoite, enlarged RBC , Schüffner’s dots Gametocyte
  • 47. More than 20 merozoites
  • 48.
  • 49. 2. Fluorescence microscope  Kawamoto technique : fluorescent staining methods : malarial parasite  Blood smear : slide : stain with Acridine orange  Nuclear DNA : green  Cytoplasmic RNA : Red  Examined under fluorescence microscope
  • 50. 3. Quantitative buffy coat (QBC)method  Sensitive method  Based on : ability of acridine orange to stain nucleic acid containing parasites  Blood : capillary tube : coated with fluorescent dye : microhaematocrit centrifugation  Buffy coat : observed under fluorescent microscope  Acridine orange stained malarial parasite : brilliant green
  • 51. 4.Serodiagnosis  IHA  ELISA  IFA 5. Rapid diagnostic kit 6. Dipstick method  An enzyme immunoassay : which detects histidine rich protein 2 Pf(HRP-2) : metabolic product produced by P.falciparum 7. Molecular diagnosis  DNA and RNA : PCR
  • 52.
  • 53.  Chloroquine  Amodiquine  Proguanil  Quinine
  • 54.