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What proportion of species are parasites? Parasitic infections I.

RNDr. et M.Res Lenka Richterová Ph.D. NRL for diagnosis of tropical parasitological infections

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What proportion of species are 2017 parasites?

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Parasites as seen by medicine Parasites as seen by medicine

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Parasites as seen by medicine Tropical tissue parasites protozoans

Plasmodium Leishmania sp. falciparum P. falciparum T. cruzi

Trypanosoma brucei Babesis sp. P. malariae P. ovale S https://www.cdc.gov/dpdx/

Intestinal parasites Tissue helmints

Cyclospora Echinococcus Trichuris spp. Microsporidia cayetanensis Entamoeba Toxocara sp. multilocularis Taenia solium O. volvulus

Enterobius Fasciola Taenia saginata vermicularis Cryptosporidium G. intestinalis Trichinella sp. hepatica Schistosoma sp. Paragonimu ssp.

https://www.cdc.gov/dpdx/ https://www.cdc.gov/dpdx/

Tropical tissue parasites protozoans

Plasmodium Leishmania sp. falciparum P. falciparum T. cruzi

Trypanosoma brucei Babesis sp. P. malariae P. ovale

https://www.cdc.gov/dpdx/

2 5/10/2021

World malaria report 2018 Plasmodium falciparum - life cycle 2016 for the first time in 20 years number of cases increased!!!

Venezuela Burundi 1mil 2018 7,2 mil 2019

S 219mil cases

S 200,5mil in Africa

S 7,51mil P. vivax

S 435 000 deths (403 000 Afrika)

http://magazine.jhsph.edu/2011/malaria/online_extras/galleries/malaria_life_cycle/ https://www.who.int/malaria/publications/world-malaria-report-2018/en/

Malaria Clinical presentation

S symptoms of uncomplicated malaria can be rather non-specific

S untreated malaria can progress to severe forms that may be rapidly (<24 hours) fatal

S most frequent symptoms include fever and chills, which can be accompanied by headache, myalgias, arthralgias, weakness, vomiting, and diarrhoea

S Other clinical features include splenomegaly, anemia, thrombocytopenia, hypoglycemia, pulmonary or renal dysfunction, and neurologic changes

Malaria Malaria RDT

S HRP – 2 S RDT antigen – P. falciparum/ P.spp Plasmodium falciparum S Microscopy – species determination, parasitemia Sequence polymorphysm S PCR – detection of sub microscopical parasitemia, mixed infections, damaged plasmodia S Aldolase Plasmodium spp. High false negativity P. ovale a P.malariae

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Malaria–RDT – HRP2 Thin and thick smear

Pfhrp2 mutace 2000 Pfhrp2 mutace 2016

Malaria Thin smear microscopy Plasmodium falciparum Plasmodium vivax

Thick smear Thin smear

Plasmodium falciparum (GI) Plasmodium falciparum (GI)

10x senzitivity Morfology

Thin smear Thin smear

Plasmodium falciparum Plasmodium vivax Plasmodium ovale Plasmodium malariae/knowlesi

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Malaria Malaria microscopy PCR

S Plasmodium falciparum S Esential for parasitaemia S Plasmodium vivax

S Detection minimum S Plasmodium ovale parasitaemia 0.001% S Plasmodium malariae S Species identification S (Plasmodium knowlesi)

Detection of mixed infections

Malaria and other parasites in blood Malaria treatment microscopy

Intermitent preventive Thick smear Thin smear ACT – first line treatment treatment in pregnancy (IPTp)

Plasmodium vivax (GI) Trypanosoma brucei spp. (GI) Plasmodium vivax (GI) Trypanosoma brucei spp. (GI)

•2010-2015 6% - 31% increase IPTp (2 doses sulfadoxine- pyrimethamine) female15-49 last pregnancy Babesia sp. (GI) Brugia malayi (GI) Babesia sp. (GI) Brugia malayi (GI) 10x sensitivity Morfology

What else can be found in blood?

S Histoplasma S 12-15 mil cases / year

S 40 000 deaths S Borelia S Over 20 species of Leishmania

S 30 species of sandfly can transmit S Candida

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Leishmaniasis Status of endemicity of :

Number of cases India/ World visceral cutaneous 2005-2013 over 60 000

http://apps.who.int/neglected_diseases/ntddata/leishmaniasis/leishmaniasis.html

Status of endemicity of Status of endemicity of visceral leishmaniasis: :

Number of cases 2016 Number of cases 2017 Eastern Mediterranean/ World 219 200 World 22 145

- Syria 2017

South-East Asia 6 176 1. Syria 47 377 Africa 6 050 58% Eastern Mediterranean 5 127 2. Afghanistan 35 184 http://apps.who.int/neglected_diseases/ntddata/leishmaniasis/leishmaniasis.html http://apps.who.int/neglected_diseases/ntddata/leishmaniasis/leishmaniasis.html 3. Pakistan 27 151 Americans 4 422 71% 4. Iraq 18 258 1990-2016 Brazil increase by 52,9%

Leishmaniasis Clinical presentation Leishmaniosis - Leishmania spp.

S Cutaneous leishmaniasis is characterized by one or more cutaneous lesions on areas where sandflies have fed. S Microscopy – scrapings of the ulcer, bone marrow, S sores can be painless or painful histological slides S Some people have swollen glands near the sores S Serology – semiquantitative haemaglutation IgG S Visceral leishmaniasis usually have fever, weight loss, and an enlarged spleen and liver

S Some patients have swollen glands. S Cultivation + Sequencing – species determination S low blood counts, including a low red blood cell count (anemia), low white blood cell count, and low platelet count.

S Some patients develop post kala-azar dermal leishmaniasis.

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Sleeping sickness- Trypanosoma Sleeping sickness gambiense,T. rhodesiense T. gambiense, T. rhodesiense

S Microscopy – ulcer, aspirate from lymph nodes, blood, CSF,

S PCR – kinetoplastid DNA Number of new cases

S Serology – 1442 (2017) Melasporol 1/20 killed 977 (2018)  95% (2000-2018) Eflornithine difficult admin. S HAT Ag for T. gambiense  2009 Nifurtimox-eflornithine  2018 Fexnidazole

Nitroimidazole derivatives - Fexinidazole

Fexinidazole Winthrop, a Sanofi-Aventis product developed with the DNDi, received a positive endorsement from the European Medicines Agency on November 15th, 2018, for use in non- European markets S (metabolit=sulfoxid a sulfon) antiparazitikum s aktivitou S Estimate 6-7 mil people, mainly in proti latin america are infected • enzyme-mediated reduction by nitroreductases to S generate cytotoxic species that cause DNA, lipid and protein damage S

S Entamoeba histolytica

S Trypanosoma brucei

S Leishmania donovani N. Of cases 2013 WHO

Chagas disease Chagas disease Trypanosoma cruzi

S Microscopy – acute – blood, CSF chronic – histological slides from biopsy

S Serology – quantitative IgG Trypanosoma cruzi

S PCR – kinetoplastid DNA

S Diagnostics – kcongenital Chagas disease

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Pneumocystosis – Clinical Pneumocystis jiroveci presentation

S These fungi are found in the lungs of mammals S dyspnea, nonproductive cough, and fever S until the host’s immune system becomes debilitated - oftentimes S Chest radiography demonstrates bilateral infiltrates lethal pneumonia S Extrapulmonary lesions occur in a minority (<3%) of S Pneumocystis jirovecii cysts are thick- patients, involving most frequently the lymph nodes, walled, rounded and approximately 5-8 spleen, liver, and bone marrow µm in size S Typically, in untreated PCP increasing pulmonary involvement leads to death.

Pneumocystosis – Toxoplasma gondii diagnostics and treatment tissue cysts: skeletal muscle, myocardium, brain, and eyes

(Tropical infectious disease 2011) S Up to 80% of population have Ab S 1) oocysts

S Diagnosis from BAL, sputum or oral wash, S 2) tachyzoits

S 3) bradyzoits (tissue cyst) S microscopy or PCR S Eating undercooked meat with bradyzoites

S Food or water contaminated with cat S Treatment: Trimetoprim-sulfamethoxazol (TMP-SMX, feces with oocysts Co-trimoxasol), Pentamidin S Blood transfusion or organ transplantation S HIV+ 21 days, non-HIV 14-21 days • serology, stained biopsy, amniotic fluid PCR S Transplacentally from mother to fetus

Toxoplasmosis Congenital toxoplasmosis clinical presentation

S from an acute primary infection acquired by S in immunocompetent persons is generally an the mother during pregnancy asymptomatic S incidence and severity vary with the S 10% to 20% with acute infection cervical trimester during which infection was acquired retionochoroiditis lymphadenopathy and/or a flu-like illness MR Lesion and edema S treatment of the mother reduce sequelae in S Immunodeficient patients - CNS disease but may have the infant (pyrimethamin + sulfadiazin + retinochoroiditis, pneumonitis, or other systemic disease acidum folinicum) S AIDS, toxoplasmic encephalitis is the most common S Ocular Toxoplasma infection – cause of intracerebral mass lesions (reactivation of retinochoroiditis, hydrocephalus, psychomotor chronic infection) retardation Calcification and mild hydrocephalus

(Tropical infectious disease 2011) (Tropical infectious disease 2011)

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Intestinal parasites Main population in Košice

0,9% Cyclospora 0,9% Trichuris spp. 1,9% Microsporidia cayetanensis Entamoeba

15,2%

Enterobius Taenia saginata vermicularis Cryptosporidium G. intestinalis Ascaris lumbricoides Enterobius vermicularis Taenia spp. Hymenolepis nana

G. Štrkolcová1, M. Goldová1, M. Maďar2 1 Ústav parazitológie, UVLF, 2Katedra mikrobiológie a imunológie, UVLF v Košiciach https://www.cdc.gov/dpdx/

Getho near Košice Parasitological examination of stool

0,31% Giardia duodenalis 0,62% 0,31% Cryptosporidium spp. 14,40% 16,10% 7,30% Ascaris lumbricoides 15,04% Trichuris trichiura

51,70% Enterobius vermicularis Hymenolepis nana

č. Ancylostomatidae

Taenia sp.

G. Štrkolcová1, M. Goldová1, M. Maďar2 1 Ústav parazitológie, UVLF, 2Katedra mikrobiológie a imunológie, UVLF v Košiciach

Parasitological examination of Parasitological examination of stool stool

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Parasitological examination of Parasitological examination of stool stool

Gomoriho-trichrom

Parasitological examination of PCR multiplex – stool parasites stool

S Stain (Trichrom, Miláček, S - Giardia lamblia Heidenheim, Calcofluor white) S - Entamoeba histolytica

Entamoeba hartmanni (TRI) Cryptosporidium parvum (MI) S - Cryptosporidium spp.

S - Blastocystis hominis

S - Dientamoeba fragilis (HHdD) Encephalitozoon sp. (CAL) S - Cyclospora cayetanensis

Diagnostical-RDT - Entamoeba histolytica

S Microscopy

Differencitation Entamoeba Entamoeba his./dis. (TRI) Entamoeba coli (TRI) histolytica from non pathogenic

Entamoeba hartmanni (TRI) Entamoeba polecki (TRI)

Endolimax nana (TRI) Iodamoeba buetschlii (TRI)

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Amoebiasis - Entamoeba histolytica Amoebiasis – Clinical presentation

S Microscopy S from asymptomatic infection (“luminal amebiasis”) Differentiation Entamoeba histolytica from nonpathogenic S to invasive intestinal amebiasis (, colitis, appendicitis, amoebs Entamoeba his./dis. (TRI) Entamoeba coli (TRI) toxic megacolon, amebomas)

S to invasive extraintestinal amebiasis (liver abscess, peritonitis, pleuropulmonary abscess, cutaneous and genital amebic lesions)

Entamoeba hartmanni (TRI) Entamoeba polecki (TRI)

Endolimax nana (TRI) Iodamoeba buetschlii (TRI) Mucosal ulcer colon Liver abscess (Tropical infectious disease 2011)

Amoebiasis - Entamoeba histolytica Amoebiasis - Entamoeba histolytica

100 000 deaths worldwide annually

S Microscopy – stool, cyst aspirate, histological slides from biopsy

S Serology – Quantitative detection of IgG Entamoeba histolytica

S PCR – stool, cyst aspirate, histological slides from biopsy 9 E.h.

Giardia intestinalis (syn. Amoebiasis - Entamoeba histolytica G.lamblia, G.duodenalis)

• The cysts are hardy and can S Microscopy survive several months in cold Differentiation Entamoeba water

histolytica from nonpathogenic Entamoeba his./dis. (TRI) amoebs • lumen of the proximal small bowel where they can be free or S PCR S Serology attached to the mucosa by a ventral sucking disk Differentiation Entamoeba S 95% extraintestinal am. histolytica from E. dispar and E. 70% activ intestinal inf. • cysts are infectious when passed moshkowskii 10% asymptomatic inf. in the stool or shortly afterward (stool, aspitrate, biopsia)

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Giardia intestinalis Giardiosis Clinical Presentation

S from asymptomatic carriage to severe diarrhoea S Most prevalent GI pathogenic estimate 200 mil./year and malabsorption

S Acute develops after an incubation period of 1 to 14 days

S usually lasts 1 to 3 weeks

S Symptoms include diarrhoea, abdominal pain, bloating, nausea, and vomiting.

S In chronic giardiasis the symptoms are recurrent

Giardia intestinalis/duodenalis/lamblia Diagnostics

S Microscopy S Giardia lamblia A (humans, primates, dogs, cats, cows, sheep, deer, rodents) S Special stain Trichrom (50-70% 1stool., 90% 3stool.) S Giardia lamblia B (humans, primates, dogs, cats, horses, beavers) S PCR detekction (for therapy monitoring) S Giardia lamblia C,D (digs) S Cysts 6-10um x 8-12um S Giardia lamblia E (cows, sheep, goats, pigs) S Trofozoites 5-15um x 9-21um (duoden st.) S Giardia lamblia F (cats) S Giardia lamblia G (pigs) S G.agilis, G. ardae, G.psittaci, G.microti, G. muris

Cryptosporidiosis - Clinical Cryptosporidium sp. presentation

S Sporulated oocysts, containing 4 S from asymptomatic infections to sporozoites, are excreted by the infected host severe, life-threatening illness through feces (contaminated water) S incubation period is an average of 7 S sporozoites are released and parasitize days epithelial cells S Watery diarrhoea is the most frequent S Two different types of oocysts are produced, symptom the thick-walled, which is commonly S chronic and more severe in excreted from the host S dehydration, weight loss, abdominal immunocompromised pain, fever, nausea and vomiting patients S and the thin-walled oocyst which is primarily involved in autoinfection S Immunocompetent 1 to 2 weeks S CD4 counts < 200/µl

(Tropical infectious disease 2011)

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http://www.cdc.gov/dpdx/ http://mikrobiologie.lf3.cuni.cz/

Tropical infectious disease 2011 S

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