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The Protozoans
Mr. F. V. Po, RMT
Taxonomic Classification of Protozoans
                                   Subphylum        Class
                                   Sarcodina        Lobosea (Amebas)
               Subphylum
               Sarcomastigophora

                                                        Class
                                    Subphylum           Zoomastigophora
                                    Mastigophora        (Hemoflagellates/
                                                        flagellates



Subkingdom   Phylum                            Class
             Ciliophora                        Kinetofragminophorea
Protozoa                                       (Ciliates)




             Phlyum                      Class
             Apicomplexa                 Sporozoa
The Intestinal Amebae
General Characteristics of Amebae
 The most important feature that separates amebae from
 other groups of Protozoans us the presence of
 pseudopods in their trophozoite form.


 Amebae have two morphologic forms: The cyst
 and the trophozoite form.

 Trophozoites are delicate, fragile and
 motile.

 The life cycles of all the intestinal amebae
 are similar.

 Replication is by binary
 fission.
Entamoeba coli




                                  Entamoeba coli
   Common Name            none
   Infective Stage        Mature cyst (Commensal; non-pathogenic)
   Diagnostic Specimen    Stool
   Mode of Transmission   Ingestion of cyst via contaminated food/ water
   Disease Caused         None (non-pathogenic)
Cysts of Entamoeba coli




            Entamobea coli cysts
Entamoeba histolytica




                              Entamoeba histolytica
  Common Name            none
  Infective Stage        Mature Quadronucleated cyst
  Diagnostic Specimen    Stool
  Mode of Transmission   Ingestion of cyst via contaminated food/ water
Trophozoites of Entamoeba histolytica
Differentiation of E. coli and E. histolytica

      Characteristics       E. histolytica cyst               E. coli cyst
        Size range              8 to 22 um                    8 to 35 um
          Shape             spherical to round            Spherical to round
     Number of nuclei          one to four                   one to eight
                                                        large, irregular shape,
       Karyosome            small and central                  eccentric
       Peripheral                                        coarse and unevenly
       Chromatin        fine and evenly distributed           distributed
        Cytoplasm            Finely granular                      coarse
       Cytoplasmic      chromatoid bars, rounded      Thin, splintered chromatoid
        inclusions                ends                             bars
                          Diffuse glycogen mass         Diffuse glycogen mass
Iodamoeba butschlii




                             Iodamoeba butschlii
 Common Name            none
 Infective Stage        Mature cyst
 Diagnostic Specimen    Stool
 Mode of Transmission   Ingestion of cyst via contaminated food/ water
Iodamoeba butschlii trophozoites, trichrome stained
Endolimax nana




                               Endolimax nana
 Common Name            none
 Infective Stage        Mature cyst
 Diagnostic Specimen    Stool
 Mode of Transmission   Ingestion of cyst via contaminated food/ water
Trophozoites of Endolimax nana
The Free Living Amebae
Acanthamoeba species




                             Acanthamoeba species
 Common Name            none
 Infective Stage        Mature cyst
 Diagnostic Specimen    CSF, brain tissue, and corneal scrapings
 Mode of Transmission   Inhalation of infective cyst or penetration through mucosa
 Disease Caused         Granulomatous Amebic Encephalitis (GAE)
Naegleria fowleri




                               Naegleria fowleri
 Common Name            none
 Infective Stage        Mature cyst
 Diagnostic Specimen    CSF
                        Penetration of ameboid trophozoite to nasal mucosa in
 Mode of Transmission   contaminated water or inhalation of cyst in dust particles
 Disease Caused         Primary Amebic Meningocencephalitis (PAM)
The Flagellates
Trichomonas vaginalis




                              Trichomonas vaginalis
  Common Name            none
  Infective Stage        Trophozoite (there is no known cyst stage)
  Diagnostic Specimen    Vaginal discharge, Urine sediments, prostatic secretion
  Mode of Transmission   Sexual intercourse
  Disease Caused         Vaginitis, urethritis (of amebic origin)
A Closer Look (Trichomonas vaginalis)
Chilomastix mesnili




                               Chilomastix mesnili
  Common Name            none
  Infective Stage        Cyst
  Diagnostic Specimen    Stool
  Mode of Transmission   Ingestion of cyst via contaminated food/ water
  Disease Caused         Not pathogenic
A Closer Look (Chilomastix mesnili)


C. mesnili cyst




C. mesnili
trophozoite
Giardia lamblia




                                Giardia lamblia
 Common Name            None
 Infective Stage        Mature cyst
 Diagnostic Specimen    Stool
                        Ingestion of mature infective cyst via contaminated food/
 Mode of Transmission   water
 Disease Caused         Traveler’s Diarrhea (Giardiasis)
A Closer Look (Giardia lamblia)
G. lamblia
Cyst
The Hemoflagellates
An Introduction to the Hemoflagellates
Leishmania species




                               Leishmania species
  Common Name            None
  Infective Stage        Promastigotes
  Diagnostic Specimen    Giemsa stained blood smear from infected area
  Mode of Transmission   Bite of infected sandfly
Leishmania species Differentiation
     Leishmania species             Vector                Disease Caused            Geographic location
                            Sandflies (Lutzomyia,   Mucocutaneous                 South American
Leishmania braziliensis     Psychodopygus)          Leishmaniasis                 Countries
                            Sandflies (Lutzomyia,   Visceral Leishmaniasis (Kala-
Leishmania donovani Complex Phlebotomus)            azar)
                                                                                  Central and South
   L. donovani chagasi                                                            America
                                                                                  Africa, India, Thailand,
   L. donovani donovani                                                           Pakistan
                                                                                  Europe, Africa,
   L. donovani infantum                                                           Northern China
                             Sandflies
Leishmania tropica Complex   (Phlebotomus)          Cutaneous Leishmaniasis/
                                                                                  Highlands of Ethiopia,
   L. aethiopica                                    Old World Leishmaniasis/      Kenya
                                                                                  USSR, Iran, Israel,
   L. major                                         Oriental Sore/                Jordan etc..
                                                                                  Afghanistan, India,
   L. tropica                                       Baghdad/ Deli Boil            Kenya etc..
Trypanosoma cruzi




                               Trypanosoma cruzi
  Common Name            none
  Infective Stage        Trypomastigotes
  Diagnostic Specimen    Blood
                         Bite from defecating reduviid bug, blood transfusion, sexual
  Mode of Transmission   intercourse, transplacental transfer
  Disease Caused         Chagas’ Disease
Miscellaneous Protozoa
Balantidium coli




                                Balantidium coli
  Common Name            None
  Infective Stage        Cyst
  Diagnostic Specimen    Stool
  Mode of Transmission   Ingestion of cyst via contaminated food/ water
  Disease Caused         Balantidiasis
Toxoplasma gondii




                               Toxoplasma gondii
  Common Name            None
  Infective Stage        Oocyst
  Diagnostic Specimen    Serum (serologic tests)
                         Ingestion of oocyst, ingestion of undercooked contaminated
  Mode of Transmission   cattle/ pig/sheep, transplacental transfer, blood transfusion
                         Toxoplasmosis/ Cerebral Toxoplasmosis/ Congenital
  Disease Caused         Toxoplasmosis
Life Cycle of Toxoplasma gondii
Blastocystis hominis




                              Blastocystis hominis
  Common Name            None
  Infective Stage        Thick walled cyst
  Diagnostic Specimen    Stool
  Mode of Transmission   Ingestion of cyst
  Treatment              Abdominal cramping, diarrher, vomiting, nausea, fever
The Plasmodia
General Characteristics of Plasmodia
  The typical life cycle of each of the Plasmodium organism involves over
  two dozen morphologic forms. However, only 6 of the most commonly
  encountered forms in human specimens will be discussed and are listed
  chronologically:


                     Ring Forms (Early Trophozoites)

                     Developing Trophozoites

                     Immature Schizonts

                     Mature Schizonts

                     Microgametocytes

                     Macrogametocytes
Identification of Plasmodia
Identification of Plasmodia
Identification of Plasmodia
Identification of Plasmodia
Identification of Plasmodia
Plasmodium vivax
Plasmodium ovale
Plasmodium malariae
A Recaptiulation
                      Plasmodium vivax Typical Characteristics at a Glance
   Relative age of infected
             RBC            Young and Immature RBC
   Appearance of Infected
             RBC            Enlarged, distorted
     Morphologic Form       Typical characteristics (based on Giemsa Stain)
          Ring Form         Delicate cytoplasmic ring measuring 1/3 RBC diameter
                            single chromatin dot
                            ring surrounds a vacuole
   Developing Trophozoite Irregular ameboid appearance
                            Ring remnants common
                            brown pigment
     Immature Schizont      Multiple chromatin bodies
                            brown pigment
      Mature Schizont       12 to 24 merozoites occupying majority of the RBC's
                            merozoites surrounded by cytoplasmic material
                            brown pigment may be present
                            large pink to purple chromatin mass surrounded by colorless to
     Microgametocytes       pale halo
                            brown pigment common
     Macrogametocytes       round to oval cytoplasm
                            eccentric chromatin mass
A Recapitulation cont’d
                  Plasmodium ovale Typical Characteristics at a Glance
  Relative age of Infected
            RBC            young and immature RBC's
  Appearance of Infected
            RBC            oval and enlarged, distorted with ragged cell membranes
    Morphologic Form       Typical characteristics (based on Giemsa Stain)
         Ring Form         Resembles that of P. vivax
                           Ring larger in size than P. vivax
                           Ring often thick and somewhat ameboid in appearance
                           Ring appearance usually maintained until late in
  Developing Trophozoite development
                           Ameboid tendencies not as evident as in P. vivax
                           Progressive dividing chromatin surrounded by cytoplasmic
    Immature Schizont      material,
                           often maintains circular shape early in development
     Mature Schizont       Parasites occupy three quarters of the RBC
                           Rosettes of an average of 8 merozoites
          Micro/
    Macrogametocytes       Similar to P. vivax, only smaller in size
A Recapitulation cont’d
                       Plasmodium malariae Typical Characteristics at a Glance
  Relative age of infected RBC's mature RBC's
  Appearance of infected RBC's normal size; no distortion (mature cells are not pliable anymore)
       Morphologic Form          Typical Characteristics (based on Giemsa stain)
            Ring Form            smaller in size than P. vivax
                                 occupies 1/6 of the RBC
                                 heavy chromatin dot
                                 vacuole may appear "filled in"
                                 pigment forms early
    Developing Trophozoite       Non-ameboid soild cytoplasm that may assume a roundish, oval
                                 band, or bar shape
                                 Cytoplasm contains coarse dark brown pigment that may mask
                                 chromatin material
                                 vacuoles are absent in mature stages
       Immature Schizont         Similar to that of P. vivax, only smaller and may contain large and
                                 dark peripheral or central granules
         Mature Schizont         6-12 merozoites arranged in rosettes or irregular clusters
                                 central arrangement of brown-green pigment may be visible
   Macro/ Microgametocyte        Similar to P. vivax, only smaller in size and pigment is usually
                                 darker and more coarse
                                 Older forms assume an oval shape
A Recapitulation cont’d
                     Plasmodium falciparum Typical Characteristics at a Glance
   Relative age of Infected RBCs   May infect cells of all ages
   Appearance of infected RBCs     Normal size; no distortion
        Morphologic Form           Typical characteristics (based on Giemsa stain)
            Ring Form              Circle configuration (one chromatin dot) or headphone
                                   configuration (two chromatin dots)
                                   Scanty cytoplasm and small vacuole
                                   Multiple rings common
                                   Accole forms common
     Developing Trophozoite        "heavy rings" common
                                   Fine pigment granules
                                   Mature forms only seen in severe infections
        Immature Schizont          Multiple chromatin bodies surrrounded by cytoplasm
                                   Only detected in severe infections
        Microgametocyte            Sausage or crescent shaped
                                   Dispersed central chromatin with nearby black pigment visible
        Macrogametocyte            Sausage or crescent shaped
                                   Compact chromatin
                                   Black pigment visible

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The protozoans

  • 2. Taxonomic Classification of Protozoans Subphylum Class Sarcodina Lobosea (Amebas) Subphylum Sarcomastigophora Class Subphylum Zoomastigophora Mastigophora (Hemoflagellates/ flagellates Subkingdom Phylum Class Ciliophora Kinetofragminophorea Protozoa (Ciliates) Phlyum Class Apicomplexa Sporozoa
  • 4. General Characteristics of Amebae The most important feature that separates amebae from other groups of Protozoans us the presence of pseudopods in their trophozoite form. Amebae have two morphologic forms: The cyst and the trophozoite form. Trophozoites are delicate, fragile and motile. The life cycles of all the intestinal amebae are similar. Replication is by binary fission.
  • 5. Entamoeba coli Entamoeba coli Common Name none Infective Stage Mature cyst (Commensal; non-pathogenic) Diagnostic Specimen Stool Mode of Transmission Ingestion of cyst via contaminated food/ water Disease Caused None (non-pathogenic)
  • 6. Cysts of Entamoeba coli Entamobea coli cysts
  • 7. Entamoeba histolytica Entamoeba histolytica Common Name none Infective Stage Mature Quadronucleated cyst Diagnostic Specimen Stool Mode of Transmission Ingestion of cyst via contaminated food/ water
  • 9.
  • 10.
  • 11. Differentiation of E. coli and E. histolytica Characteristics E. histolytica cyst E. coli cyst Size range 8 to 22 um 8 to 35 um Shape spherical to round Spherical to round Number of nuclei one to four one to eight large, irregular shape, Karyosome small and central eccentric Peripheral coarse and unevenly Chromatin fine and evenly distributed distributed Cytoplasm Finely granular coarse Cytoplasmic chromatoid bars, rounded Thin, splintered chromatoid inclusions ends bars Diffuse glycogen mass Diffuse glycogen mass
  • 12. Iodamoeba butschlii Iodamoeba butschlii Common Name none Infective Stage Mature cyst Diagnostic Specimen Stool Mode of Transmission Ingestion of cyst via contaminated food/ water
  • 14. Endolimax nana Endolimax nana Common Name none Infective Stage Mature cyst Diagnostic Specimen Stool Mode of Transmission Ingestion of cyst via contaminated food/ water
  • 16. The Free Living Amebae
  • 17. Acanthamoeba species Acanthamoeba species Common Name none Infective Stage Mature cyst Diagnostic Specimen CSF, brain tissue, and corneal scrapings Mode of Transmission Inhalation of infective cyst or penetration through mucosa Disease Caused Granulomatous Amebic Encephalitis (GAE)
  • 18. Naegleria fowleri Naegleria fowleri Common Name none Infective Stage Mature cyst Diagnostic Specimen CSF Penetration of ameboid trophozoite to nasal mucosa in Mode of Transmission contaminated water or inhalation of cyst in dust particles Disease Caused Primary Amebic Meningocencephalitis (PAM)
  • 20. Trichomonas vaginalis Trichomonas vaginalis Common Name none Infective Stage Trophozoite (there is no known cyst stage) Diagnostic Specimen Vaginal discharge, Urine sediments, prostatic secretion Mode of Transmission Sexual intercourse Disease Caused Vaginitis, urethritis (of amebic origin)
  • 21. A Closer Look (Trichomonas vaginalis)
  • 22. Chilomastix mesnili Chilomastix mesnili Common Name none Infective Stage Cyst Diagnostic Specimen Stool Mode of Transmission Ingestion of cyst via contaminated food/ water Disease Caused Not pathogenic
  • 23. A Closer Look (Chilomastix mesnili) C. mesnili cyst C. mesnili trophozoite
  • 24. Giardia lamblia Giardia lamblia Common Name None Infective Stage Mature cyst Diagnostic Specimen Stool Ingestion of mature infective cyst via contaminated food/ Mode of Transmission water Disease Caused Traveler’s Diarrhea (Giardiasis)
  • 25. A Closer Look (Giardia lamblia) G. lamblia Cyst
  • 27. An Introduction to the Hemoflagellates
  • 28.
  • 29.
  • 30. Leishmania species Leishmania species Common Name None Infective Stage Promastigotes Diagnostic Specimen Giemsa stained blood smear from infected area Mode of Transmission Bite of infected sandfly
  • 31. Leishmania species Differentiation Leishmania species Vector Disease Caused Geographic location Sandflies (Lutzomyia, Mucocutaneous South American Leishmania braziliensis Psychodopygus) Leishmaniasis Countries Sandflies (Lutzomyia, Visceral Leishmaniasis (Kala- Leishmania donovani Complex Phlebotomus) azar) Central and South L. donovani chagasi America Africa, India, Thailand, L. donovani donovani Pakistan Europe, Africa, L. donovani infantum Northern China Sandflies Leishmania tropica Complex (Phlebotomus) Cutaneous Leishmaniasis/ Highlands of Ethiopia, L. aethiopica Old World Leishmaniasis/ Kenya USSR, Iran, Israel, L. major Oriental Sore/ Jordan etc.. Afghanistan, India, L. tropica Baghdad/ Deli Boil Kenya etc..
  • 32. Trypanosoma cruzi Trypanosoma cruzi Common Name none Infective Stage Trypomastigotes Diagnostic Specimen Blood Bite from defecating reduviid bug, blood transfusion, sexual Mode of Transmission intercourse, transplacental transfer Disease Caused Chagas’ Disease
  • 34. Balantidium coli Balantidium coli Common Name None Infective Stage Cyst Diagnostic Specimen Stool Mode of Transmission Ingestion of cyst via contaminated food/ water Disease Caused Balantidiasis
  • 35. Toxoplasma gondii Toxoplasma gondii Common Name None Infective Stage Oocyst Diagnostic Specimen Serum (serologic tests) Ingestion of oocyst, ingestion of undercooked contaminated Mode of Transmission cattle/ pig/sheep, transplacental transfer, blood transfusion Toxoplasmosis/ Cerebral Toxoplasmosis/ Congenital Disease Caused Toxoplasmosis
  • 36. Life Cycle of Toxoplasma gondii
  • 37. Blastocystis hominis Blastocystis hominis Common Name None Infective Stage Thick walled cyst Diagnostic Specimen Stool Mode of Transmission Ingestion of cyst Treatment Abdominal cramping, diarrher, vomiting, nausea, fever
  • 38.
  • 40.
  • 41. General Characteristics of Plasmodia The typical life cycle of each of the Plasmodium organism involves over two dozen morphologic forms. However, only 6 of the most commonly encountered forms in human specimens will be discussed and are listed chronologically: Ring Forms (Early Trophozoites) Developing Trophozoites Immature Schizonts Mature Schizonts Microgametocytes Macrogametocytes
  • 50. A Recaptiulation Plasmodium vivax Typical Characteristics at a Glance Relative age of infected RBC Young and Immature RBC Appearance of Infected RBC Enlarged, distorted Morphologic Form Typical characteristics (based on Giemsa Stain) Ring Form Delicate cytoplasmic ring measuring 1/3 RBC diameter single chromatin dot ring surrounds a vacuole Developing Trophozoite Irregular ameboid appearance Ring remnants common brown pigment Immature Schizont Multiple chromatin bodies brown pigment Mature Schizont 12 to 24 merozoites occupying majority of the RBC's merozoites surrounded by cytoplasmic material brown pigment may be present large pink to purple chromatin mass surrounded by colorless to Microgametocytes pale halo brown pigment common Macrogametocytes round to oval cytoplasm eccentric chromatin mass
  • 51. A Recapitulation cont’d Plasmodium ovale Typical Characteristics at a Glance Relative age of Infected RBC young and immature RBC's Appearance of Infected RBC oval and enlarged, distorted with ragged cell membranes Morphologic Form Typical characteristics (based on Giemsa Stain) Ring Form Resembles that of P. vivax Ring larger in size than P. vivax Ring often thick and somewhat ameboid in appearance Ring appearance usually maintained until late in Developing Trophozoite development Ameboid tendencies not as evident as in P. vivax Progressive dividing chromatin surrounded by cytoplasmic Immature Schizont material, often maintains circular shape early in development Mature Schizont Parasites occupy three quarters of the RBC Rosettes of an average of 8 merozoites Micro/ Macrogametocytes Similar to P. vivax, only smaller in size
  • 52. A Recapitulation cont’d Plasmodium malariae Typical Characteristics at a Glance Relative age of infected RBC's mature RBC's Appearance of infected RBC's normal size; no distortion (mature cells are not pliable anymore) Morphologic Form Typical Characteristics (based on Giemsa stain) Ring Form smaller in size than P. vivax occupies 1/6 of the RBC heavy chromatin dot vacuole may appear "filled in" pigment forms early Developing Trophozoite Non-ameboid soild cytoplasm that may assume a roundish, oval band, or bar shape Cytoplasm contains coarse dark brown pigment that may mask chromatin material vacuoles are absent in mature stages Immature Schizont Similar to that of P. vivax, only smaller and may contain large and dark peripheral or central granules Mature Schizont 6-12 merozoites arranged in rosettes or irregular clusters central arrangement of brown-green pigment may be visible Macro/ Microgametocyte Similar to P. vivax, only smaller in size and pigment is usually darker and more coarse Older forms assume an oval shape
  • 53. A Recapitulation cont’d Plasmodium falciparum Typical Characteristics at a Glance Relative age of Infected RBCs May infect cells of all ages Appearance of infected RBCs Normal size; no distortion Morphologic Form Typical characteristics (based on Giemsa stain) Ring Form Circle configuration (one chromatin dot) or headphone configuration (two chromatin dots) Scanty cytoplasm and small vacuole Multiple rings common Accole forms common Developing Trophozoite "heavy rings" common Fine pigment granules Mature forms only seen in severe infections Immature Schizont Multiple chromatin bodies surrrounded by cytoplasm Only detected in severe infections Microgametocyte Sausage or crescent shaped Dispersed central chromatin with nearby black pigment visible Macrogametocyte Sausage or crescent shaped Compact chromatin Black pigment visible