Introduction to Protozoan parasites

2. ....much of which is parasitic The majority of eukaryotic life is protozoan...

3. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar Phylogeny of protozoan parasites kinetoplastids

4. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum us Naegleria Giardia Insect vector ToxoplasmaPlasmodium brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar Kinetoplast - deﬁning feature of all kinetoplastids kinetoplastids

5. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum us Naegleria Giardia Insect vector ToxoplasmaPlasmodium brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar Kinetoplast - deﬁning feature of all kinetoplastids kinetoplastids

6. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum us Naegleria Giardia Insect vector ToxoplasmaPlasmodium brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar Kinetoplast - deﬁning feature of all kinetoplastids kinetoplastids Lukes, Euk. Cell, 2002

7. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum us Naegleria Giardia Insect vector ToxoplasmaPlasmodium brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar kinetoplastids Lukes, Euk. Cell, 2002 Hajduk, FASEB, 1993 Kinetoplast - deﬁning feature of all kinetoplastids

8. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar deﬁning features of alveolates

9. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar deﬁning features of alveolates

10. The Inner Membrane Complex (IMC)  underlies the parasite plasma membrane IMC: ﬂattened vesicles + cytoskeleton Toxoplasma

11. ER Mt DG PM IMCPM IMC The Inner Membrane Complex (IMC)  underlies the parasite plasma membrane IMC: ﬂattened vesicles + cytoskeleton Toxoplasma

12. ER Mt DG PM IMCPM IMC The Inner Membrane Complex (IMC)  underlies the parasite plasma membrane IMC: ﬂattened vesicles + cytoskeleton Gliding machinery: - MyoA - MLC1 - GAP45 - GAP50 - GAP40 Toxoplasma

13. ER Mt DG PM IMCPM IMC The Inner Membrane Complex (IMC)  underlies the parasite plasma membrane IMC: ﬂattened vesicles + cytoskeleton Cytoskeletal elements: - Microtubules - Alveolins - GAPMs - ISPs Gliding machinery: - MyoA - MLC1 - GAP45 - GAP50 - GAP40 Toxoplasma

14. Spatial-temporal organization of the IMC is critical for Plasmodium differentiation MD Lazarus et al, J Cell Sci 121:1937-‐49; 2008 LG Tilney, unpublishedMD Lazarus et al, J Cell Sci 121:1937-‐49; 2008 Parasitophorous vacuole (PV) Parasite Plasma Membrane Inner Membrane Complex (IMC) Merozoite Trophozoite Schizont

15. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar deﬁning features of apicomplexa

19. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar deﬁning features of apicomplexa > 5000 species of parasites

24. Interpreting parasite life-cycles

25. Interpreting parasite life-cycles • What is the deﬁnitive host (sexual reproduction)?

26. Interpreting parasite life-cycles • What is the deﬁnitive host (sexual reproduction)? • What is the intermediate host (asexual reproduction)?

27. Interpreting parasite life-cycles • What is the deﬁnitive host (sexual reproduction)? • What is the intermediate host (asexual reproduction)? • Can there be more than one host species?

28. Interpreting parasite life-cycles • What is the deﬁnitive host (sexual reproduction)? • What is the intermediate host (asexual reproduction)? • Can there be more than one host species? • How are the host species geographically distributed?

29. Interpreting parasite life-cycles • What is the deﬁnitive host (sexual reproduction)? • What is the intermediate host (asexual reproduction)? • Can there be more than one host species? • How are the host species geographically distributed? • How does transmission occur?

30. Interpreting parasite life-cycles • What is the deﬁnitive host (sexual reproduction)? • What is the intermediate host (asexual reproduction)? • Can there be more than one host species? • How are the host species geographically distributed? • How does transmission occur? • What cell type(s) does the parasite infect?

31. Interpreting parasite life-cycles • What is the deﬁnitive host (sexual reproduction)? • What is the intermediate host (asexual reproduction)? • Can there be more than one host species? • How are the host species geographically distributed? • How does transmission occur? • What cell type(s) does the parasite infect? • What are the major developmental stages?

32. gut gut Redblood cells Sporozoites intohost Sexualstages invector Intestinal epithelium Salivary glands Intestinal epithelium Merozoites Toxoplasma Crypto- sporidium Plasmodium Theileria Tick Mosquito Cat(Muscle, neurons) Mammalian Host Hepatocytes Lymphocytes Anycell (Bradyzoites) Roos, Science 309:72-74 (2005) Apicomplexa share similar life-cycle strategies

33. Toxoplasma life-cycle Hunter & Sibley. Nature Rev. Micro. 2012

34. Toxoplasma life-cycle Hunter & Sibley. Nature Rev. Micro. 2012 Intermediate host! (asexual reproduction)

35. Toxoplasma life-cycle Hunter & Sibley. Nature Rev. Micro. 2012 initial infection leads rapid cell invasion and parasite replication from Gary Ward! Univ. of Vermont Intermediate host! (asexual reproduction)

36. Dinkorma Ouologuem 14

40. 15

41. 15

45. Toxoplasma life-cycle Hunter & Sibley. Nature Rev. Micro. 2012 Asexual reproduction occurs in intermediate hosts

46. Toxoplasma life-cycle

47. Toxoplasma life-cycle

48. Toxoplasma life-cycle Sexual reproduction occurs in Felidae (deﬁnitive host)

49. Toxoplasma life-cycle Sexual reproduction occurs in Felidae (deﬁnitive host)

50. 18

51. 18 Kreuder, et al., J. Wildlife Diseases, 2003

52. 18 Conrad, et al., IJP, 2005 Kreuder, et al., J. Wildlife Diseases, 2003

53. 18 Conrad, et al., IJP, 2005 Kreuder, et al., J. Wildlife Diseases, 2003

54. 18 Conrad, et al., IJP, 2005 Kreuder, et al., J. Wildlife Diseases, 2003 Torey & Yolken, TrePar, 2013

57. Cryptosporidium: sexual and asexual reproduction occur in the same tissue of the same host very resilient! common contaminant of water

58. ID50 = 100-300 cysts Cryptosporidium = ‘hidden spore’ NEJM, March 1995

59. Cryptosporidium = ‘hidden spore’ NEJM, March 1995

60. Cryptosporidium = ‘hidden spore’ ~400,000 people affected! 69 people died (93% had HIV/AIDS)

62. • GEMS set out to identify infectious causes of mild to severe cases of pediatric diarrhea in developing countries ! • 7 study sites in Africa and SE Asia; 3 year study! • ~10,000 children with MSD and ~13,000 controls (0-5yrs)! • multiplex diagnostics leveraging a variety of molecular and standard microbiological assays.

63. The Lancet, July 2013

64. The Lancet, July 2013

67. Plasmodium life-cycle

70. Malaria

71. Malaria Ranks as one of the ‘big three’ in total mortality for infectious diseases

72. Malaria Ranks as one of the ‘big three’ in total mortality for infectious diseases 220 million people infected in 2010

73. Malaria Ranks as one of the ‘big three’ in total mortality for infectious diseases 220 million people infected in 2010 most are in sub-Saharan Africa

74. Malaria Ranks as one of the ‘big three’ in total mortality for infectious diseases 220 million people infected in 2010 most are in sub-Saharan Africa over 600,000 deaths in 2010 alone

75. Malaria Ranks as one of the ‘big three’ in total mortality for infectious diseases 220 million people infected in 2010 most are in sub-Saharan Africa over 600,000 deaths in 2010 alone 90% of these are in children under age 5

76. Malaria Ranks as one of the ‘big three’ in total mortality for infectious diseases 220 million people infected in 2010 most are in sub-Saharan Africa over 600,000 deaths in 2010 alone 90% of these are in children under age 5 global prevalence has dropped 25% since 2000

77. Why is malaria so deadly? Miller, et al., Science, 1994 vol. 264: 1878-1883.

78. Why is malaria so deadly?

82. P. vivax

85. Leishmaniasis

86. cutaneous Leishmaniasis

87. cutaneous mucocutaneous Leishmaniasis

88. cutaneous mucocutaneous visceral Leishmaniasis

89. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar kinetoplastids African Trypanosomes" T. bruci rhodesiense (East African Sleeping Sickness) - only 2% of disease, acutely progressive! T. bruci gambiense (West African Sleeping Sickness) - accounts for 98% of disease

90. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar kinetoplastids African Trypanosomes" T. bruci rhodesiense (East African Sleeping Sickness) - only 2% of disease, acutely progressive! T. bruci gambiense (West African Sleeping Sickness) - accounts for 98% of disease

91. tsetse ﬂy humans! buffaloes! cattle T. brucei (African sleeping sickness)

97. Only 1% of tsetse ﬂies are infected

98. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar kinetoplastids American Trypanosomiasis" T. cruzi (Chagas disease)

99. Protozoa alveolates Fecal-oral Foodborne Mode of transmission Coccidians Cryptosporidium Excavates LeishmaniaTrypanosoma ApicomplexaCiliates DinoﬂagellatesHeterolobosia Euglenoza Fornicata major subgroup Phylum Genus Naegleria Giardia Insect vector ToxoplasmaPlasmodium 'brain-eating amoeba' 'traveler's diarrhea' 'traveler's diarrhea' malariaChagas disease African sleeping sickness Kala azar kinetoplastids American Trypanosomiasis" T. cruzi (Chagas disease)

100. T. cruzi is transmitted by reduviid bugs

101. T. cruzi life-cycle

102. T. cruzi life-cycle

Introduction to Protozoan parasites

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