1. Parasitology - protosoology (protosoa), helmintology(worms), enthomology(insects)
Laboratory diagnosis: - life cycle of parasits, material v laboratory dg
Protozoa – intestinal, genital, urinary, blood, tissue
Worms - Helmints: Nematodes, Cestodes,Trematodes
Ectoparasits: louse, ticks, flies – important as vectors

2. Diagnosis Problematic, not ususal outside endematic areas
Nonspecific clinical manigestation
eosinophilia in helmintoses – not constant sign
Importance of history – personal, travellers, social, economic, food, therapy
Conditions for successful dg:
- think on parasitosis,
- correct sampling – right sample, with good method at right time, sent in appropriate conditions to the lab that is able to identify the parasit, good interpretation:
Knowledge of life cycle is principal

3. Life cycle of parasits -terminology
Complex life cycle – key to diagnosis
Host – hosts
Definitive host – parasit finishes the growing cycle and is becoming adult in it
Not typical host - parasit cannot develop in it
Transient host – larves are developing in it but sexual multiplication is not performed
Helmints : egg - larva - (cyste) – adult worm Protozoa :trofozoit - motile, cyste – non motile sexual multiplication - zygota asexual- schizonts, sporogons

4. Patogenesis Chronic infections
Protosoa – In strong immunity reaction – imunopathologic signs – symptoms hypersensitivity, cross reacting antigens (tissue devastation) and autoimunity
Worms – mechanic, big, metabolism – direct injury (elefantiosis)
- imunity and inflamation – indirect immunopathological reaction

5. Imunity production of IgE
Worms stimulates CD4Th lymfocytes – they secrete IL-4, IL-5
ADCC – via eosinophils and IgE – elimination of worms – basic protein in granulesof eosinophils is toxic for worms
CD4 - activation of macrophages – elimination of parasits = Production of granuloms in organs, fibrosis, tissue devastation
I.c. parasits activate cytotoxic lymphocytes – interference of disseminations

6. How parasits escape human immunity
= chronicity
In-borne nonspecific mechanisms – faibly efficient
Specific mechanisms – antigenic mimicry – the surface of the parasit is covered with host antigens, komplexes of Ag-Ab, i.c.localisation of parasits, antigenic variation during infection
- size, motility, cover of albunim, extreme production of antigen imunity overload, imnosuression

7. Therapia Less structutres Toxic
Knowledge of the life cycle of parasit is primary
Eucaryotic cells of parasits – toxicita – selective toxicity can be achieved by changes of the molecule by metabolising of it in the body of parasit, acumulation of the molecule in the body of parasit
Chronicity – long term therapy
Tissue localisation, encystation
Socioeconomic conditions, climatic factors

8. Stool for parasitological examination
Macroscopy – blood, mucous, adult worms
Microscopy - native smear - FS, iode – motility, eggs of helmints, cysts of protosoa, ery, leu
Concentration methods – separation of cysts of protosoa and eggs of helmints from other material in the stool
Stainning - identification – smear of native stool + hematoxylin eosin, trichrome

9. Other material acc.to clinical manifestation
Perianal - Enterobius vermicularis
Sigmoidoscopy - Entamoeba histolytica
Duodenal aspiration - Giardia lamblia
Biopsia of abscesse of liver - Entamoeba histolytica
Sputum - Ascaris lumbricoides, Strongyloides,
Urine - Schistosoma
Urogenital sample - Trichomonas
Blood - (malaria, trypanosomiosis, leishmaniosis, filariosis) smear, thick drop - staining acc. Giemsa, HeO,
Serum

10. Protosoa
Amoeba - Entamoeba histolytica, Entamoeba coli, Naegleria fowleri, Acanthamoeba, Endolimax nana
Flagelata - Giardia lamblia, Trichomonas vaginalis, Leishmania, Trypanosoma
Ciliata - Balantidium coli
Coccidia a Sporosoa - Cryptosporidium, Blastocystis, Microsporidia, Plasmodia, Babesia,, Toxoplasma

11. Nematodes -worms
Enterobius vermicularis, Ascaris lumbricoides, Toxocara canis,cati, Trichuris trichiura, Ancylostoma duodenale, Strongyloides stercoralis, Trichinella spiralis, Wuchereria bancrofti, Dracunculus medinensis – aesculape´s bat
Nonsegmented body, adult worms living in the GIT - diagnosis: identification of eggs in the stool (morphology of eggs)
Filariae – thin worms parasiting in eye, skin, tissues, transmitted by insects. Larval stage - microfilariae penetrate to blood and are transmitted by suckling insect

12. Cestodes Head - scolex, segmented body
- Hermafrodit, male and female organs are present in every segment - dif.dg.
- They have not GIT, absorbtion of food.
- Complex life cycle with transient host (sometimes - human – larval stage of cysticercosis, echinococcosis)
Taenia solium, Taenia saginata, Diphylobotrium latum, Echinococcus granulosus, Echinococcus multiloculare Hymenolepsis nana

13. Trematodes - Usually hermafrodits (exception - Schistosoma)
Need transient host (hosts)
Fasciolopsis, Clonorchis, Paragonimus, Schistosoma

14. Nematodes -worms
Enterobius vermicularis, Ascaris lumbricoides, Toxocara canis,cati, Trichuris trichiura, Ancylostoma duodenale, Strongyloides stercoralis, Trichinella spiralis, Wuchereria bancrofti, Dracunculus medinensis – aesculape´s bat
Nonsegmented body, adult worms living in the GIT - diagnosis: identification of eggs in the stool (morphology of eggs)
Filariae – thin worms parasiting in eye, skin, tissues, transmitted by insects. Larval stage - microfilariae penetrate to blood and are transmitted by suckling insect

15. Enterobius vermicularis
Definitive host - human
transient - none
dg. – perianal sample – microscopy of eggs
fecal oral transmission - autoinfection

16. Enterobius vermicularis

17. Ascaris lumbricoides Definitive host: human
Larva migrans: intestin, colon - muc.membrane - blood- lung - cough - mouth - colon
dg. Egg in stool
Infection via contaminated food
Symptoms acc.to localisation of the larva

18. Ascaris lumbricoides

19. Dracunculus

20. Cestodes Head - scolex, segmented body
- Hermafrodit, male and female organs are present in every segment - dif.dg.
- They have not GIT, absorbtion of food.
- Complex life cycle with transient host (sometimes - human – larval stage of cysticercosis, echinococcosis)
Taenia solium, Taenia saginata, Diphylobotrium latum, Echinococcus granulosus, Echinococcus multiloculare Hymenolepsis nana

21. Taenia solium Definitive: pig transient: rat
incidental: human in small intestin
dg. serology
Contaminated food

23. Echinococcus granulosus
definitive: dog
transient:sheep
incidental: human
dg serology
transmission: cyste in meat
infection: mechanic pressure from expanded cyst, prolonged growing, rupture of the cyst and dissemination

24. Echinococcus granulosus

25. Intestinal,brain and UG protosoa
entamoeba histolytica
entamoeba coli
giardia lamblia
naegleria fowleri, acanthamoeba
trichomonas vaginalis

26. Entamoeba histolytica
definitive: human
dg.trofosoits in stool, serology
dysenteria – diarhea with blood

27. Naegleria fowleri, Acantamoeba
Free Living in water
Human (via nose)
dg.microscopy in CSF- identification of invasive strains)
disease: purulent peracute meningitis

29. Trichomonas vaginalis
Definitive: human
transient: none
dg. Cultivation - microscopy trophosoit - from vagina, urine
Sexual transmission
Therapy of both (all) partners

31. Blood and tissue protosoa
Plasmodium vivax – tertiana
Plasmodium ovale – tertiana
Plasmodium malariae – quartana
Plasmodium falciparum – maligna tertiana
Toxoplasma gondii
Pneumocystis carinii – jiroveci
Leishmania donovani – kala azar
Trypanosoma brucei gambiense – sleeping disease
rhodesiense

32. Toxoplazma gondii Definitive: cat transient: rat incidental: human
dg. serology KFR, IgA, IgG, IgM
transmission: food borne, hand, annimal
disease: - intrauterine primoinfection generalised lymfadenopathy, encystation in organs – abortion, eye……..

34. Life cycle
Cat swallow tissue cyst (in food) or oocyst (from excrements) – infection of epitelial cells of small intestin – asexual and sexual multiplication – formation of oocysts – excremetion during 1-2 weeks – cyst is nonsporulated = not infectious – outside the body after 1-5 days cysts sporulate – infectious during months, resistent to desinfection, freezing, drying, heating to 70 dg C for10 mins.

36. Clinical manifestation
Sabin – meningoencefalitída
- Pinkerton a Henderson – tyfoidné príznaky
Siim – generalised lymphadenopathy -
lymphocyto-monocytes proliferation in periferal blood
gained toxoplasmosis
gained or reactivated toxoplasmosis in IDS
In borne – congenital toxoplasmosis
Eye toxoplasmosis

37. Gained toxoplasmosis Usually asymptomatic
10% - 20% cervical lymphadenopathy + flue like symptoms
Clinicallly selfliminting – reactivation in IDS, in gravidity

38. Gained toxoplasmosis in IDS
Primary infection or reactivation of anamnestic infection
CNS , myocarditis, pneumonia
AIDS pacients – encephfalitis, intracerebral lesions

39. Congenital toxoplasmosis
In acute primoinfection of mother during pregnancy
Symptoms depends on the lenghth of gravidity during primoinfection
Therapy can decrease the symptomatology
Acute diagnosis is important
New borne can have subclinical symptoms – without therapy usually getting worse sy i.u. toxoplasmosis – hydrocefalus, calcifications in brain and liver, cataracta, microcephalus

40. Eye toxoplasmosis Ofthen asymptomatic untill the 2nd-3rd decenium
Symotoms – rupture of the cyst in eye, tachysoits and bradysoits are released
Chorioretinitis – unilateral – after gained infection – bilateral – after i.u. infection

41. Toxoplasma gondii protozoa i.c. parasit
Infiects different warm-blood annimals
Cat is host for sexual stages of Toxoplasmy gondii (schisonts) – main source
3 stages – tachysoits (trofosoits) – quick multiplication and destruction of the invaded tissue – bradysoits – slowly multipliing in tissue cysts – sporosoits (male and female gamonts) in oocysts – in cat excrements

42. developmental stages of Toxoplasma gondii
Neonsporulating adn sporulating cyst from cat stool
Tachysoits free and i.c.
Bradysoits in tissue cysts

43. Infection in men
1) food borne with inappropriately boiled food containing tissue cysts
2) transplantacentarly
3) accidentally by innoculation of tachysoits
4) swallowing of oocysts (sandy playgrounds., contaminated hands)
5) kontaminated trasfusion or transplantation

44. Pathogenesis Swallowing of the cyst (bradysoit) or oocyst (sporosoit)
Releasing of microorganism
Invasion to the small intestin epitelium
Dissemination and multiplication intracelularly
Death of infected cells, releasing od tachysoits that invade other cells
Reaction of immunity system, change of tachy to bradysoits, formation of tissue cysts (in muscles, heart, brain)
Reactivation during IDS – rupture of cysts, releasing of parasits

45. Direct proof Seldom positive Staining acc. Giemsa Immunofluorescence
ELISA for antigen detection
Tissue cultures
Innoculation of mouses
PCR

46. Laboratory results IgG IgM specific antibodies:
– – without serological proof
– probable acute infection (+IgA from the same
sample) or false positive IgM
reaction (repetition of IgG and IgM
from neuw sample – no changes)
probable infection 6 – 12 mnths ago.
– probable infection 1 year ago
KFR – total antibody detection

47. CFR titres above 8 are suspect for infection dynamicity
– increase of the titre after 14 days - acute – maps decrease of the infection after therapy and possible reactiovation of the infection CFR antibodies are life long persistent

48. Serology of toxoplasma gondi in pregnancy
I. trimestre – test for IgM antibodies – detection of i.u. infection – positivity – indicate recetn infection current IgA positivity – acute infection – therapy of mother and screening of the baby
IgG antibodies positive (seldome with IgM positivity) – not indicating acute infection, can result in reactivation of possible tissue cyst, thah has no relation to pregnancy and cannot be eliminated by therapy
Increase of IgG antibodies – reactivation of past infection without threat of fetus health

49. Antenatal diagnosis of inborne toxoplasmosis
Acute infection in pregnant mother
Therapia
Ultra sonography
Amnionic fluid + fetal blood: PCR innoculation do mouses and tissue cultures
Fetal blood: Toxo IgM and IgA, activity of liver ensyms

50. Newborne infection dg Isolation from placenta, umbilicus leucocytes
Clinical and laboratory test for tissue injury detection
Špecific antibodies – newborne serum CSF – mother serum
IgG,M,A IgM,IgA
PCR
tachysoit

51. Sequelae of toxoplasma infection in women
primary infection – possibility of fetal death – lethal infection of the fetus
intrauterine infection and birth of ill baby
Previous infection with cysts in tissue of gential tract – problems with conception and succesfull pregnancy (habitual abortions)

52. Plasmodium-malariae, falciparum
Definitive host:Anopheles
Transient host: human, monkey
dg.microscopy thick drop
Transmission insect bite
disease: malaria acc.to the rate of schizogonia- clinically as fever attacks - tercianna, quartana,