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GAB 23 october 2007

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GAB 23 october 2007

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  1. GAB 23 october 2007

  2. Parasitology, 2007? • M. Lontie • MCH, Leuven

  3. Ocular micrometer disk • each objective must be calibrated with reference material • can be roughly checked with a counting chamber, with RBCs ...

  4. Eosinophilia > 10% With helminths, insects (myasis), not with protozoa excepting Isosporabelli and Dientamoebafragilis (with pinworms?)

  5. Auto-infectious Enterobius vermicularis Hymenolepis nana Strongyloides stercoralis

  6. Threats • Global warming • El Nino(little child in Spanish, arriving around Christmas). • Dams • Diama dam in Senegal and large outbreak of Schistosoma mansoni infections. 1996. Kongs et al. Trop Med Int Health. 191-8. • The Lancet. 2001. 570-1.

  7. IatrogenicHepatitis C in Egypt and mass treatment of schistosomiasis • Rao M. et al. 2002. BMC Infect Dis: 29. Nile delta of Egypt: 1950-1980. Injections of tartar emetic (antimony potassium tartrate) with inadequately sterilized needles and syringes.

  8. M. Wéry, 1995.

  9. Kovats R. et al. 2003. The Lancet . El Nino and health

  10. Kovats R. et al. 2003. The Lancet . El Nino and health

  11. Specific gravities • S.G. of Zn-sulfate 33 %: 1.180 • S.G. of formol-solution 10 %: 1.019 • S.G. of ether: 0.714 • S.G.of parasites: Ancylostoma 1.055; Giardia 1.060; Entamoebahistolytica (coli) and Endolimaxnana 1.065 - 1.070; Ascaris 1.110; Trichuris 1.150, Chilomastixmesnili 1.180; Ascaris (unfertilized) 1.200 (Bailenger, 1965).

  12. Entamoeba histolytica • Trophozoite (magna variety) in faeces. Diameter approximately 30 m. Nucleus with typical fine chromatin picture (iron-hematoxylin stain).

  13. Entamoeba histolytica • Trophozoite (magna variety) in faeces. Diameter approximately 30 m. Nucleus with typical fine chromatin picture and central karyosome (Lugol stain).

  14. Entamoeba histolytica • Trophozoite (minuta variety) in faeces. Diameter approximately 15 m. Nucleus with typical fine chromatin picture (iron-hematoxylin stain).

  15. Entamoeba histolytica immature cyst mature cyst trophozoite

  16. Entamoeba histolytica • Cyst in faeces with three visible nuclei. In the nucleus at the right we clearly see the central karyosome (Lugol stain).

  17. Entamoeba histolytica • Cyst in faeces with two visible nuclei. In the nucleus at the left we clearly see the central karyosome (Lugol stain).

  18. Entamoeba histolytica • Cyst in faeces with one visible nucleus with a central karyosome (Lugol stain).

  19. Entamoeba histolytica • Cyst in faeces with one visible nucleus with a central karyosome (Lugol stain).

  20. Entamoeba histolytica • Cyst in faeces with one visible nucleus with a central karyosome (Lugol stain).

  21. Entamoeba histolytica • Cyst in faeces with one visible nucleus with a central karyosome (Lugol stain).

  22. Entamoeba histolytica • Cyst with one visible nucleus and one cylindrical chromatoidal body (Lugol stain).

  23. Entamoeba histolytica • Cyst with two nuclei and one cylindrical chromatoidal body (Lugol stain).

  24. Entamoeba histolytica • Cyst with one visible nucleus and one cylindrical chromatoidal body (Lugol stain).

  25. Entamoebahistolytica - dispar • Entamoebapolecki • Entamoebahartmanni (small race E.histolytica) • Entamoebahistolytica Laredo strain • Entamoebadispar • non pathogenic strains grow betweeen 20 and 37°C, pathogenic only at 37°C • isoenzyme analysis: only 9 zymodemes are pathogenic

  26. Entamoebahistolytica - dispar • PCR, isoenzyme analysis, and antigen detection (JCM, 1998, 449). • Monoclonal antibodies (JCM, 2001, 716). • ITG-Antwerp: PCR under evaluation (T. Vervoort): fecal material in 3.5% formalin.

  27. Entamoeba coli • Large cyst (> 20m) in faeces with four visible nuclei (Lugol stain).

  28. Dientamoeba fragilis • Two nuclei. • “The unflagellated human flagellate “. • Only (very labile) trophozoites, no cysts. • Questionable enteric pathogen. • Doxycycline, paromomycin, metronidazole (Sanford et al., 2005).

  29. Courtesy CDC

  30. Dientamoeba fragilis • SAF fixative (sodium acetate acetic acid formalin) and iron hematoxylin stain have replaced the PVA (polyvinyl alcohol fixative with HgCl2) and trichrome stain.

  31. Sodium acetate acetic acid formalin (SAF) • Sodium acetate 1.5 g • Acetic acid, glacial 2.0 ml • Formaldehyde, 37-40 % 4.0 ml • Distilled water 92.0 ml

  32. Dientamoeba fragilisIn saline (unstained).

  33. Dientamoeba fragilisIn saline (unstained).

  34. Dientamoeba fragilisIn saline (unstained).

  35. Dientamoeba fragilis • In The Netherlands. • In 247 unpreserved stool specimens: none. • In 247 SAF-preserved stool specimens: 24. • (Mank T. 1997. Thesis) Stained with hematoxylin.

  36. Dientamoeba fragilis • In Brussels, Belgium. • SAF-preserved stool specimens used. • D. fragilis (6.3%) and G. lamblia (7.1%) in 448 patients. Stained with hematoxylin. Vandenberg O. et al. 2006. Int J Infect Dis:221, 2.

  37. Dientamoeba fragilis MCH, Leuven • (2005-2007) • 6 + / 160 SAF Stained with hematoxylin.

  38. Blastocystis hominis • Cyst in faeces (Hematoxylin stain).

  39. AntoonvanLeeuwenhoekGiardialamblia1681

  40. Giardia lamblia • Trophozoite with two nuclei and several flagella (May-Grünwald-Giemsa stain)

  41. Giardia lamblia: antigen detection by IF and ELISA • monoclonal antibodies: Merifluor (MERIDIAN) (Cryptosporidium and Giardia). • 8/9 Giardia ELISAs are OK. (JCM, 1998, 1338). • Triage parasite panel (BIOSITE) useful. (JCM, 2000, 3337; JCM, 2001, 334). • one ELISA almost as sensitive as two microscopic examinations. (Mank T. 1997).

  42. Antigen detection • Good sensitivity and specificity • In combination with conventional tests • Problem = COST (Triage = $ 19.44/test)

  43. Leishmaniasis • Leishmaniainfantum, Leishmaniadonovani, … in macrophages of man. • Female sandflies: Phlebotomus spp., Lutzomia spp.,... • Cutanous, visceral leishmaniasis (Kala-azar). • Fever, hepatosplenomegaly, pancytopenia. • Serology - parasitological examination of bone marrow aspiration. • (A. Van Gompel et al., T v G, 1997)

  44. Leishmania sp. • Ovoid small (2-6m) intracellular parasite in a bone marrow aspirate. The typical rod shaped kinetoplast is seen besides the nucleus (May-Grünwald-Giemsa stain).