24TH HELMINTHOLOGICAL DAYS 2018 abstract book... · 2018. 10. 11. · 24th Helminthological Days...

Czech Society for Parasitology

Charles University, Faculty of Science

24TH HELMINTHOLOGICAL DAYS

2018

Programme & Abstracts

Editors: Jana Bulantová & Tomáš Macháček

Prague 2018

24th Helminthological Days

Organized by: Czech Society for Parasitology, Helminthological Section

Venue & date: Hotel Kouty, Rejčkov 20, Czechia, 7–11 May 2018

Organising committee: Tomáš Macháček, Jana Bulantová, Libuše Turjanicová, Oldřich

Vondráček, Martin Majer (Department of Parasitology, Faculty of

Science, Charles University, Prague, Czechia)

Conference website: https://hd2018.natur.cuni.cz

Conference email: [email protected]

© Czech Society for Parasitology, 2018

Cover illustration © Lucie Jedličková, 2018

ISBN 978-80-7444-057-1

Programme

1

CONFERENCE PROGRAMME

Invited talk: 25 min + 5 min for discussion

Regular talk: 10 min + 5 min for discussion

Poster talk: 2 min (+ individual discussion at the poster)

BSc/MSc or PhD Presentations included in a student com-

petition evaluated by a conference com-

mittee. The winners will be announced

and awarded during the closing ceremony.

MONDAY, MAY 7

15:00 – 18:00 Arrival and registration of participants

18:00 – 00:00 Get-together-evening with barbecue party

TUESDAY, MAY 8

08:00 – 08:45 Breakfast

09:00 – 09:15 Opening ceremony (T. Macháček)

Session I Biochemistry & diagnostics (Chairman: T. Macháček)

09:15 – 09:45 P. Matoušková, I. Vokřál, L. Raisová Stuchlíková, J. Lamka, B. Szo-

táková, L. Skálová: Metabolism of anthelmintics and drug resistance

in helminths (invited talk)

09:45 – 10:00 I. Vokřál, P. Matoušková, H. Danková, B. Szotáková, L. Skálová:

Efflux transporters in Haemonchus contortus – role in drug resistance

development

10:00 – 10:15 K. Skulinová, J. Novák, L. Kolářová, M. Kašný: Identification of im-

munogenic proteins in excretory-secretory products of Toxocara canis

larvae (PhD)

10:15 – 10:30 M. Ježková, R. Leontovyč, M. Kašný: In silico design, construction

and evaluation of multi-epitope chimeric antigens for serodiagnosis of

fasciolid flukes (BSc/MSc)

10:30 – 10:45 V. Šteiger, P. Horák, M. Kašný: Molecular determination of Tricho-

bilharzia spp. DNA in the body fluids of their hosts (BSc/MSc)

10:45 – 11:15 Coffee break

Programme

2

Session II Novel approaches in helminthology (Chairman: R. Leontovyč)

11:15 – 11:30 P. Roudnický, T. Loja, D. Potěšil, Z. Zdráhal, M. Gelnar, M. Kašný:

Laser microdissection and mass spectrometry: protein analysis of

strictly selected tissue from Eudiplozoon nipponicum (PhD)

11:30 – 11:45 O. Vondráček, P. Talacko, R. Leontovyč, J. Bulantová, P. Horák: Pro-

teomic analysis of helminth tissues using microdissection – from cry-

osections to raw data (PhD)

11:45 – 12:00 L. Turjanicová, D. L. Nguyen, A. van Diepen, C. H. Hokke, P. Horák,

L. Mikeš: Glycomic analysis of developmental stages of the neuro-

tropic schistosome Trichobilharzia regenti (PhD)

12:00 – 12:15 N. Reslová, V. Huvarová, L. Škorpíková, M. Kašný, P. Králík: Future

of multiplexing (PhD)

12:30 – 13:15 Lunch

Session III Proteases of helminths (Chairman: J. Dvořák)

13:30 – 14:00 A. Leontovyč, J. Dvořák: Schistosomes and their smart weapons –

proteases (invited talk)

14:00 – 14:15 L. Konečný, M. Kašný, L. Mikeš, P. Horák: Characterisation of se-

lected peptidases from Trichobilharzia regenti cercariae (BSc/MSc)

14:15 – 14:30 K. Peterková, P. Horák, M. Kašný: Fascioloides magna digestive pep-

tidases - cathepsin L, B and D: gene expression and tissue localization

of their transcripts (BSc/MSc)

14:30 – 14:45 L. Jedličková, H. Dvořáková, J. Dvořák, L. Ulrychová, V. Žárský, M.

Kašný, L. Mikeš: Cysteine and aspartic peptidases in the blood-

feeding monogenean Eudiplozoon nipponicum

14:45 – 15:00 M. Havelková, H. Dvořáková, L. Jedličková, L. Mikeš: Kunitz-like

peptidase inhibitors of the blood-feeding monogenean Eudiplozoon

nipponicum (BSc/MSc)


15:30 – 16:00 S. Morand: Amphibians and their helminth parasites: macroecological

patterns and knowledge gaps (invited talk)

Session IV Helminths in silico (Chairman: M. Kašný)

16:00 – 16:15 J. Bulantová, V. Krčmářová, P. Horák: Feeding preferences in two

distinct species of avian schistosomes of the genus Trichobilharzia

16:15 – 16:30 R. Leontovyč, N. Young, M. Kašný, R. Gasser, P. Horák: Compara-

tive transcriptomic analysis shows molecular mechanisms used by

visceral and neurotropic avian schistosomes (PhD)

16:30 – 16:45 J. Vorel, M. Jankůjová, J. Oppelt, F. Pardy, P. Roudnický, J. Ilgová,

L. Mikeš, M. Gelnar, M. Kašný: Monogeneans in the pores (PhD)

Programme

3

Poster Session Poster talks (Chairman: L. Turjanicová)

Individual discussion at the posters

16:45 – 18:00 P1 M. Čenková, R. Kuchta: Life-cycle of Schyzocotyle acheilognathi

(Cestoda: Bothriocephalidea)

P2 D. Dimunová, L. Raisová Stuchlíková, H. Navrátilová, P. Ma-

toušková: The role of the UDP-glycosyltransferases in the metabolism

of anthelmintics in Haemonchus contortus

P3 M. Matoušková, M. Bruňanská, M. Oros, J. Nebesářová: The lo-

calization of glycogen in the mature spermatozoa of some caryophyl-

lidean tapeworms

P4 D. Matušková, T. Šulcová, E. M. Boateng, L. Ulrychová, M. Asa-

hina, C. Bařinka, J. Dvořák: Worms as relevant biological models for

investigation of Glutamate carboxypeptidase 2

P5 J. Pankrác, B. Vojtková, V. Kolářová, J. Sádlová, L. Šefc: In vivo

imaging techniques for parasitology research

P6 L. Škorpíková, N. Reslová, E. Pozio, M. Kašný: Differentiation of

eight Trichinella species using a high resolution melting assay

P7 D. Uhrovič, M. Oros, T. Scholz: New insights into the scolex mor-

phology of monozoic cestodes (Caryophyllidea) from the Nearctic region

18:00 – 18:45 Dinner

19:00 – 00:00 Campfire evening

WEDNESDAY, MAY 9

08:00 – 08:45 Breakfast

Session V Host-parasite interactions (Chairman: P. Horák)

09:00 – 09:30 P. Hyršl: Entomopathogenic nematodes and insect immunity

(invited talk)

09:30 – 09:45 T. Macháček, K. Immig, P. Horák: Dynamics of immune cells in the

CNS of mice after infection by Trichobilharzia regenti (PhD)

09:45 – 10:00 B. Šmídová, T. Macháček, P. Horák: Production of nitric oxide in mice

infected by Trichobilharzia regenti (Schistosomatidae) (BSc/MSc)

10:00 – 10:15 M. Majer, T. Macháček, P. Horák: Does the neurotropic helminth in-

fluence the peripheral immune response? (BSc/MSc)

10:15 – 10:30 N. Bernardová, M. Chanová: Development of neuromotor disorders

during experimental neurotoxocariasis (PhD)


Programme

4

Session VI Helminths under the microscopes (Chairman: J. Bulantová)

11:00 – 11:15 L. Vlčková, P. Horák: Interactions between Trichobilharzia regenti

and duck definitive host in the terminal phase of infection: histological

evaluation (BSc/MSc)

11:15 – 11:30 K. Svobodová, P. Horák: Characterization of egg and miracidium ul-

trastructures of two schistosome species with regard to their different

life strategies (BSc/MSc)

11:30 – 11:45 D. Barčák, H. Sehadová, M. Oros: CLSM observation of plerocercoid

morphology: interspecific differences of two species of Dibothrio-

cephalus (Cestoda)

12:00 – 12:45 Lunch

13:00 – 17:00 Half-day trip to Želiv

17:30 – 18:15 Dinner

18:30 – 19:00 Workshop with Olympus: Figure Maker “hands on” session 1



THURSDAY, MAY 10

08:00 – 08:45 Breakfast

Session VII Epidemiology (Chairman: T. Scholz)

09:00 – 09:30 L. Kolářová: Alveolar echinococcosis in the Czech Republic – omni-

present? (invited talk)

09:30 – 09:45 M. Oros, D. Barčák, D. Uhrovič: Occurrence of the zoonotic parasites

Anisakis spp. in Atlantic herring (Clupea harengus) marketed in Slovakia

9:45 – 10:00 Z. Pokrupová, J. Bulantová, P. Horák: The past and the present of avian

schistosomes and cercarial dermatitis in the Czech Republic (BSc/MSc)

10:00 – 10:15 N. Kmentová, M. Van Steenberge, S. Koblmüller, F. M. Bukinga, T. Mu-

limbwa N'sibula, P. M. Mulungula, M. Gelnar, M. P. M. Vanhove: Failure

to diverge connected with low rate of molecular evolution in an African

freshwater parasitic flatworm (Monogenea, Diplectanidae) (PhD)


Programme

5

Session VIII Diversity of helminths (Chairman: M. Oros)

10:45 – 11:00 A. Faltýnková, S. Georgieva, H. Mazanec, J. Roháčová, K. Skírnis-

son, A. Kostadinova: Trematodes in sub-Arctic lakes in Iceland

11:00 – 11:15 H. Mazanec, J. Roháčová, S. Georgieva, K. Skírnisson, A. Faltýnko-

vá: Digeneans of the family Strigeidae Railliet, 1919 from Iceland:

morphology and molecular characterisation (BSc/MSc)

11:15 – 11:30 C. P. de Oliveira, T. Scholz, J. L. Luque, A. Jones: A synoptic review

of cladorchiids (Digenea: Paramphistomoidea) parasites of freshwater

fishes in South America and prospects (PhD)

11:30 – 11:45 K. Vyčítalová, B. Koubková, A. Šimková: Diversity of diplozoid spe-

cies (Monogenea, Diplozoidae) parasitizing the gills of endemic cy-

prinids from Mediterranean area (PhD)

11:45 – 12:00 L. Li, L. Lü, S. A. Nadler, D. I. Gibson, L.-P. Zhang: A new insight

into the classification of the Ascaridoidea

12:15 – 12:45 Lunch

Session IX Ecology and evolution I. (Chairman: R. Kuchta)

13:15 – 13:45 M. Jirků: Natural grazing: how to combine nature management, big

herbivires and their parasites (invited talk)

13:45 – 14:00 P. Heinclová, A. Born-Torrijos, M. Soldánová: The effect of direct

predation by fish on cercarial populations of selected trematode spe-

cies (Digenea) (BSc/MSc)

14:00 – 14:15 T. Vyhlídalová, M. Soldánová: Production and periodicity in the

emergence of cercariae of Diplostomum spp. (Digenea) in the lakes of

temperate and subarctic regions (BSc/MSc)

14:15 – 14:30 M. Soldánová, A. Born-Torrijos, J. Schwelm, G. S. van Beest, T.

Vyhlídalová, E. H. Henriksen, R. Knudsen, R. Kristoffersen, P.-A.

Amundsen: Evaluating the role of trematode cercariae in the energe-

tics of a subarctic ecosystem


Session X Ecology and evolution II. (Chairman: M. Soldánová)

15:00 – 15:15 R. Kuchta, A. Choudhury, T. Scholz: Asian fish tapeworm: the most

successful invasive parasite in freshwaters

15:15 – 15:30 M. Pravdová, M. Ondračková, J. Kolářová, K. Grabicová, T. Randák,

P. Jurajda: Association between pharmaceuticals and parasite infec-

tion in natural brown trout (Salmo trutta) populations (PhD)

15:30 – 15:45 V. Krasnovyd, A. Šimková: Impact of the fish hybridization on the

parasite community: specificity, diversity and co-evolution (PhD)

15:45 – 16:00 M. Benovics, Y. Desdevises, R. Šanda, J. Vukić, A. Šimková: Evolu-

tionary history of host-specific Dactylogyrus parasites (Monogenea)

in the selected regions of peri-Mediterranean (PhD)

Programme

6

16:00 – 17:30 Presentation of conference partners

18:00 – 00:00 Closing ceremony and social event

FRIDAY, MAY 11

08:00 – 08:45 Breakfast

09:00 – 10:00 Departure of participants

7

ABSTRACTS

The abstracts, ordered alphabetically according to the presenting author, are published as

received from the authors who are fully responsible for the content. No editing or corrections

were made except for minor changes in formatting to keep the layout unified.

Abstracts

8

CLSM observation of plerocercoid morphology:

interspecific differences of two species of Dibothriocephalus (Cestoda)

D. Barčák1,2, H. Sehadová3, M. Oros1

1 Institute of Parasitology, Slovak Academy of Sciences, Košice, Slovakia

2 Institute of Parasitology, Biology Centre, Czech Academy of Sciences, v.v.i.,

České Budějovice, Czechia 3 Institute of Entomology, Biology Centre, Czech Academy of Sciences, v.v.i.,

České Budějovice, Czechia

Tapeworms of the genus Dibothriocephalus Lühe, 1899, commonly known as “broad fish

tapeworms”, are etiological agents of zoonotic intestinal disease called “dibothriocephalosis”

(syn. diphyllobothriosis). The present study was targeted on the plerocercoids (larval stages)

of two European species, which differ from each other in the structure of frontal glands, per-

haps due to their different site preferences in fish intermediate host. Larvae of Dibothrioce-

phalus latus (Linnaeus, 1758) were obtained from musculature of European perch (Perca flu-

viatilis), whereas those of D. dendriticus (Nitzsch, 1824) were isolated from the body cavity

of brown trout (Salmo trutta fario). As specific procedure for visualisation of the frontal

glands of Dibothriocephalus larvae is not currently known, several antibodies against α tubu-

lin, β tubulin, serotonin, FMRF amide and synapsin were used to identify the gland cells by

using confocal laser scanning electron microscopy (CLSM). Additionally, the effect of

hot/cold fixatives on later reactivity of internal structures was tested in the former species. In

D. latus, frontal glands and their ducts connected with the tegument were visualized by anti-α

and anti-β tubulin antibodies in larvae fixed with ‘cold’ fixative. The area of gland cells sur-

rounds the inverted scolex in the first third of the plerocercoid body, and ducts of gland cells

are weakly visible as narrow empty tubes. Anti-serotonin, anti-FMRF amide, and anti-

synapsin antibodies interacted solely with the nervous system. In plerocercoids fixed in hot

formalin, a very conspicuous “network” tagged by both anti-β tubulin and anti-serotonin anti-

bodies surrounds bothria on everted scolex from both lateral sides. These structures were con-

nected with tegument by two types of numerous duct-like structures, (a) the narrow fibres,

and (b) slightly wider fibres with enlargement in their course, often close to the basal mem-

brane of the tegument. Plerocercoids of D. dendriticus were all fixed in cold fixative and their

frontal gland cells were visualised solely by anti-β tubulin antibodies and grouped as a small

field on each side of bothria. Anti-serotonin antibodies were taged on the surface of the tegu-

ment and formed circular lines. Based on our preliminary results, it seems that hot fixation

may be beneficial for visualization of particular structures in plerocercoids of Dibothrio-

cephalus. However, the limitations of the present study was a low number of D. latus speci-

mens and lack of hot-fixed D. dendriticus plerocercoids.

Acknowledgements: The study was financially supported by the Czech Science foundation (pro-

ject No. P505/12/G112), SAS-MOST Joint Research Project (no. 2016/7), and APVV 15 0004.

Abstracts

9

Evolutionary history of host-specific Dactlogyrus parasites (Monogenea)

in the selected regions of peri-Mediterranean

M. Benovics1, Y. Desdevises2, R. Šanda3, J. Vukić4, A. Šimková1

1 Department of Botany and Zoology, Masaryk University, Brno, Czechia

2 Observatoire Océanologique de Banyuls/Mer, UPMC University Paris, Banyuls/Mer, France 3 Department of Ecology, Charles University, Prague, Czechia

4National Museum, Prague, Czechia

Parasite genus Dactylogyrus is generally known for the remarkable species diversity and high

level of host-specificity. The high number of species (more than 900) the most probably

emerged from the multitude of the host species, belonging to Cyprinidae, the largest freshwa-

ter fish family. Due to the high host-specificity we can expect that evolutionary history of

Dactylogyrus is intimately linked with the phylogeny of the cyprinids and reflects their bio-

geography and historical dispersion.

Dactylogyrus parasites were collected over years 2014 to 2018 from endemic cyprinids in the

selected peri-Mediterranean regions. In total, representatives of 113 cyprinid species belong-

ing to 24 genera were processed. 93 of all investigated host species were parasitized by Dac-

tylogyrus. From 69 identified Dactylogyrus species 18 were potentially new for science (in-

cluding D. omenti sp. n. collected from Aulopyge huegelii). The highest Dactylogyrus species

diversity was found in the representatives of the Barbus, Chondrostoma, Luciobarbus, Rutilus

and Squalius genera. The phylogenetic reconstruction based on sequence data of partial ribo-

somal SSU combined with partial ribosomal LSU revealed several strongly supported groups

within Dactylogyrus of Southern European Peninsulas and suggest multiple origin of the Dac-

tylogyrus in Iberian Peninsula. The results of phylogenetic analyses combined with the com-

putation of genetic distances and application of Mantel test revealed that however, the geo-

graphically adjacent populations are more similar also on molecular level, the recent distribu-

tion of the Dactylogyrus species is highly influenced by introduction of non-native cyprinid

species into new regions.

Applying cophylogenetic methods we found the significant coevolutionary signal between

phylogenies of endemic Balkan and Apennine Dactylogyrus and their cyprinid hosts. Strong

evolutionary link was detected between D. vistulae and its respective hosts what suggest that

host-switch played important role in the evolutionary history of Dactylogyrus.

This study was financially supported by Czech Science Foundation project No. 15-1938S and

since 2018 partially by project No. P505/12/G112.

Abstracts

10

Development of neuromotor disorders during experimental neurotoxocariasis

N. Bernardová, M. Chanová

Institute of Immunology and Microbiology, 1st Faculty of Medicine, Charles University and

General University Hospital in Prague, Prague, Czechia

Toxacara canis is globally distributed parasite of canids. As a paratenic host, wide range of

vertebrates including humans can be infected by accidental ingestion of infectious stage of

this parasite. Even though seroprevalence in human population is high (up to 36 % in some

locations), the number of diagnosed symptomatic cases known as larval toxocariasis is low.

Clinical symptoms are caused by complex, still insufficiently examined effect of migrating

larvae which persist in tissues for a long period. According to the localization of larvae in the

body, several types of larval toxocariasis are classified. Neurotoxocariasis (or cerebral toxoca-

riasis) as potential result of central nervous system (CNS) invasion is the most serious. How-

ever, the development of neurotoxocariasis is extremely rare and the presence of larvae in

CNS is generally considered as asymptomatic. Fatal neurologic disorders appear regularly

only in late-phase chronic experimental murine toxocariasis after very long time without

symptoms. Some authors call asymptomatic presence of larvae (in mice, as well as in human)

in question. They suppose ongoing pathological interactions in this phase, leading to the de-

velopment of chronic neurodegenerative and neuropsychiatric diseases. The aim of our study

was to evaluate the manifestations preceding severe neurological consequences in experimen-

tally infected mice. Mice infected by various doses of Toxocara canis larvae were tested regu-

larly for their motor abilities. Although mice did not show any obvious signs of disabilities in

common activities under standard conditions, they were slowly developing difficulties with a

balance and limb coordination which progressed finally into severe neuromotor problems.

These results confirm that the long-term presence of Toxocara canis larvae even in mice

healthy at first sight is definitely not asymptomatic.

Acknowledgements: The work was supported by Charles University (SVV 260 369, Progress Q25).

Abstracts

11

Feeding preferences in two distinct species of avian schistosomes

of the genus Trichobilharzia

J. Bulantová, V. Krčmářová, P. Horák

Department of Parasitology, Faculty of Science, Charles University, Prague, Czechia

Family Schistosomatidae contains important representatives of deadly human parasites. They

are known as gonochoristic blood dwelling trematodes of threadlike body, feeding mainly on

red blood cells. Complicated digesting machinery, including participation of important prote-

ases, helps to the worms effectively exploit blood meal as a source of nutrients. At the same

time, a huge amount of potentially toxic iron compounds (hem) abundantly present in ery-

throcytic haemoglobin must be detoxified and recycled within the body of parasites, or deliv-

ered out of their organism. Blood meal is used as a source of nutrients also in the visceral avi-

an schistosome Trichobilharzia szidati. After penetration of cercariae into the definitive bird

host, schistosomula migrate from the skin intravascularly through the lungs to the venules of

small intestine, where they mature. During the whole migratory route, the main source of food

is blood. Based on our ultrastructural examination of T. szidati intestinal content, digestion of

haemoglobin and its conversion to the crystals of hemozoin on lipid droplets inside the gut

lumen is similar to that described for the human schistosome Schistosoma mansoni. Another

Trichobilharzia species, T. regenti, feeds on the blood, but only in the adult stage. Interesting-

ly, migration of young schistosomula is closely connected not to the blood vessels but to the

nervous system of vertebrate hosts. Spinal cord is used as a highway from featherless skin of

legs (the site of usual cercarial penetration) to the definitive localization of adult worms in the

nasal mucosa of natural avian hosts. Thus, also nourishment of T. regenti schistosomula is

completely different than that of T. szidati. Moreover, thanks to ultrastructural in situ observa-

tion, we can conclude, that the type of intestinal content of T. regenti schistosomula depends

on actual position of each individual. The worms migrating under meninges feed on abundant

immune cells infiltrating the area, but schistosomula migrating through the white matter feed

mainly on myelin sheaths of neurons. Representatives of the genus Trichobilharzia are im-

portant for human health due to their ability to cause cercarial dermatitis. Peculiarities in mi-

gratory route and feeding preferences of schistosomula stimulated us to perform a complex

comparison of transcriptomic data from cercariae of both species that undergo the same way

of life, and schistosomula which for particular species use a completely different source of

nutrients (see the contribution by R. Leontovyč et al.).

Acknowledgement: The study was supported by Czech Science Foundation (Grant Nos. 13-

29577S and 18-11140S), by the project “Centre for Research of Pathogenicity and Virulence of

Parasites” (no. CZ.02.1.01/0.0/0.0/16_019/0000759) funded by European Regional Develop-

ment Fund and Ministry of Education, Youth and Sports of the Czech Republic and by Charles

University institutional support (PROGRES Q43, UNCE 204017, and SVV 244-260432/2017).

Abstracts

12

Life-cycle of Schyzocotyle acheilognathi (Cestoda: Bothriocephalidea)

M. Čenková, R. Kuchta

Institute of Parasitology, Biology Centre, Czech Academy of Sciences and Faculty of Science,

University of South Bohemia, Branišovská 31, 370 05 České Budějovice, Czechia

The life-cycle of the Asian fish tapeworm Schyzocotyle acheilognathi (Yamaguti, 1934),

which is a worldwide distributed invasive species, was studied under experimental conditions.

Adult tapeworms were obtained from Syphysodon aequitasciatus (Cichlidae) provided by a

private breeder in the Czech Republic in June 2017. A total of 210 tapeworms were found, but

only 5 of them were gravid, i.e. containing eggs. The eggs were isolated from gravid proglot-

tids and placed into Petri dishes with tap water. Hatching was observed after 2–4 days in the

laboratory at temperature of 22°C. One or two hatched ciliated larvae (coracidia) were added

to small chambers with a single copepod Diacyclops bicuspidatus (Claus, 1857) each for 24

hours. After exposure to coracidia, copepods were transferred to large beakers, maintained at

18°C and examined regularly after two weeks under light microscope. 6,19 % (194/12) of

copepods were infected. Infected copepods were added to 6 young goldfish Carassius auratus

(L.) and 2 young carp Cyprinus carpio L. and examined 3 and 7 months after exposure to co-

pepods. However, no tapeworms were found in any of the experimental fish.

Acknowledgements: This study was partly supported by the Czech Science Foundation (pro-

ject No. P505/12/G112) and the Institute of Parasitology, BC CAS (RVO: 60077344).

Abstracts

13

A synoptic review of cladorchiids (Digenea: Paramphistomoidea) parasites

of freshwater fishes in South America and prospects

C. P. de Oliveira¹, T. Scholz², J. L. Luque¹, A. Jones³

¹ Departament of Animal Parasitology, Federal Rural University of Rio de Janeiro,

Rio de Janeiro, Brazil ² Institute of Parasitology, Biology Centre of the Czech Academy of Sciences,

České Budějovice, Czechia ³ Creigiau Llwydion, Heol y Maes, Coelbren, Neath, SA10 9PT, UK

Cladorchiids are trematodes of the superfamily Paramphistomoidea that occur mainly in

freshwater fishes in the tropics. These parasites in freshwater have a very patchy geographical

distribution, having been found in North and South America, Asia, Africa, ex-USSR and Aus-

tralia. However, the majority of species occur in the Neotropical Region, especially in the

Amazon River basin. These parasites were first reported from fish in South America by Di-

esing (1836). At present, 32 species are known in 22 genera, being the second richest family

in this region. The majority of species was described from fishes in Brazil, whereas only a

few records are available from other South American countries including Argentina, Colom-

bia, French Guiana and Peru. Cladorchiids have been reported from five orders and 13 fami-

lies of fishes. Of these, the Serrasalmidae (Characiformes) is by far the most commonly in-

fected family. The validity of some species should be confirmed because their species de-

scriptions were insufficient or were based on poor-quality specimens. No molecular data were

available for any of the cladorchiids in South American fishes. Recently, a large number of

cladorchiids were collected by the present authors in Brazil and Peru, which made it possible

to describe three new species and to erect two new genera to accommodate them. In addition,

molecular data have been obtained for 12 species of cladorchiids to assess phylogenetic rela-

tionships of these trematodes.

Acknowledgments: This work was supported by the Czech Science Foundation (project

P505/12/G112) and by the “Ciências sem fronteiras Brazilian program-visiting researcher

modality granted to José L. Luque (No. 135/2012) (Stay of Tomáš Scholz in Brazil at the Uni-

versidade Federal Rural de Rio de Janeiro). CP was supported by a postgraduate fellowship

from CNPq.

Abstracts

14

The role of the UDP-glycosyltransferases in the metabolism

of anthelmintics in Haemonchus contortus

D. Dimunová, L. Raisová Stuchlíková, H. Navrátilová, P. Matoušková

Department of Biochemical Sciences, Faculty of Pharmacy, Charles University,

Hradec Králové, Czechia

Haemonchus contortus, a hematophagous gastrointestinal parasite, endanger predominantly

small ruminants, such as sheep and goats, has a great ability to develop resistance to anthel-

mintic drugs. Therefore, it is necessary to understand the mechanism of the resistance. The

effects of anthelmintics can be reduced by xenobiotic-metabolizing enzymes by decreasing

the concentration of the active drugs in parasite cells. UDP-glycosyltransferases (UGTs), im-

portant enzymes in the metabolism of xenobiotics and eobiotics, could protect the helminth

from toxic action of anthelmintics by their metabolism to inactive glycosides.

In the previous study, the adult nematodes were cultivated in the medium with anthelmintics

and metabolites were determined by using UHPLC/MS/MS analysis. These results show that

albendazole, ricobendazole and flubendazole underwent glycosidations caused by UDP-

glycosyltransferases. Except sex-differences in anthelmintics metabolism, more glycosylated

metabolites were observed in IRE (benzimidazol resistant) strain than ISE (sensitive) strain of

H. contortus (Raisová 2018). This analysis confirmed the connection between anthelmintics

resistance and their metabolism. Using quantitative PCR, the differences in the constitutive

expression of UGTs between IRE and ISE strains were analyzed. Several enzymes from the

UGTs superfamily, e.g. UGT368B2, were significantly more expressed in IRE strain than ISE

strain. The enhanced expression of biotransformation enzymes, could lead to increased rate of

anthelmintics metabolism. Based on these results, we have selected several UGTs to study

their specific role in metabolism. Functional characterization of recombinant UGTs will be

realized in baculovirus-infected insect cells. Revealing the role of these enzymes could con-

tribute to a more detailed understanding of the resistance in Haemonchus contortus. Raisová

Stuchlíková L. et al. (2018) IJP: Drugs and Drug Resistance. 8(1), 50-58.

Acknowledgements: This project was supported by Czech Science Foundation (Grant No. 17-

11954Y) and by Charles University (PRIMUS/17/SCI/4).

Abstracts

15

Trematodes in sub-Arctic lakes in Iceland

A. Faltýnková1, S. Georgieva1,2, H. Mazanec1,3, J. Roháčová1,3, K. Skírnisson4, Kostadinova A.1

1 Institute of Parasitology, Biology Centre of the Czech Academy of Sciences,

České Budějovice, Czechia 2 Science Park, Marine Zoology Unit, Cavanilles Institute of Biodiversity and Evolutionary

Biology, University of Valencia, Valencia, Spain 3 University of South Bohemia in České Budějovice, Faculty of Science,

České Budějovice, Czechia 4 Laboratory of Parasitology, Institute for Experimental Pathology, Keldur,

University of Iceland, Reykjavík, Iceland

In recent years, there have been discoveries of a surprisingly diverse spectrum of trematodes

in the Arctic and sub-Arctic regions, including diplostomids from Iceland, but also other

trematodes from Norway and North America. This is challenging the assumption that north-

ern regions are generally poor in pathogens, as they are characterised by low species diversity

and short, simple food webs because of harsh climatic conditions. As parasites are now re-

cognised as important players in the ecosystems, and are sensitive to environmental changes

in the Arctic, northern ecosystems are thus promising model systems. In a project aimed on

biodiversity in Iceland (2018–2020) we will test whether sub-Arctic lakes in Iceland harbour

trematodes of a comparable diversity to those of lakes in Norway. Therefore, we will provide

baselines for trematode diversity and abundance and a quantitative framework for further re-

search on community composition and structure in host-trematode systems in the Arctic food

webs. The project builds on previous data obtained from Iceland and continues in sampling of

pulmonate snails (Radix balthica, Gyraulus laevis) and fishes (Gasterosteus aculeatus, Salmo

trutta, Salvelinus alpinus) from lakes in and near to Reykjavík, Lake Hafravatn and Lake

Mývatn; additionally, birds (Anatidae) will be obtained from local Icelandic authorities.

Trematodes obtained will be processed morphologically and genetic markers (cox1, nad1, the

complete ITS1-5.8S-ITS2 gene cluster, and 28S rRNA gene) will be used for molecular char-

acterisation. Parasite abundance and community structure data will be analysed, host-parasite

interactions will be modelled as bipartite interaction networks. Data obtained in previous

years indicate that trematodes (of the families Diplostomidae, Echinostomatidae, Plagiorchi-

idae, Strigeidae) using birds as definitive hosts are the most abundant in freshwater, while

trematodes (Allocreadiidae) using fishes as definitive hosts are limited to few species only.

The trematode diversity in Iceland is mostly influenced by migratory bird hosts nesting or

staging in Iceland, which are overwintering in e.g. Northern Europe, Netherlands or Great

Britain. Radix balthica, the only snail being abundant in Iceland, is used as the main interme-

diate host by trematodes.

This study was supported by the Research Fund of the University of Iceland and the Czech

Science Foundation (18-18597S).

Abstracts

16

Kunitz-like peptidase inhibitors of the blood-feeding monogenean

Eudiplozoon nipponicum

M. Havelková, H. Dvořáková, L. Jedličková, L. Mikeš


Proteins containing Kunitz domains are usually protease inhibitors binding reversibly to their

target, mostly serine proteases. They are generally small proteins around 6 – 10 kDa, but also

multidomain Kunitz proteins exist. Three disulphide bonds are characteristic to their pepti-

dase-binding loop and the target specificity is mostly given by a single amino acid in the ac-

tive site. They appear widely in plants and animals, where they take part in regulation of

many physiological proteolysis-based processes, and also participate in interactions with

pathogens. These protease inhibitors are used in self-defence mechanisms, e.g., in anemone

mucus or plant seeds, and also make constituents of animal venoms.

We focus on Kunitz-like proteins of parasites, especially blood feeders, which have to deal

with specific aspects of their feeding strategy. Namely, they must fight immune and tissue

recovering responses of the hosts including blood coagulation. Numerous serine proteases are

involved in coagulation pathways, complement cascade, and inflammation, and sanguivorous

parasite species employ a variety of inhibitors to modulate these processes in the host. Kunitz

proteins were described from helminths such as Ancylostoma, Fasciola hepatica or Echino-

coccus granulosus, and also from blood feeding arthropods such as Ixodes scapularis, Boo-

philus microplus, and Simulium vittatum. Therefore it is obvious that this kind of protease

inhibitors is widely spread among parasites.

We study Kunitz-like proteins in Eudiplozoon nipponicum, a monogenean parasite from gills

of common carp. As the parasite constantly feeds on fish blood, the inhibitors of coagulation

are obligatory compounds of the feeding process. Several Kunitz proteins were identified in

the transcriptome of this parasite and one of them has been characterized as an effective inhi-

bitor of factor Xa of the coagulation cascade. We performed a bioinformatic analysis of all

E. nipponicum Kunitz proteins and compared their primary structures, elucidating their speci-

ficity toward peptidases. Selected inhibitors will be produced as recombinants and their effect

on fish physiology and immunity will be studied in detail, as well as their potential in biomed-

icine.

Acknowledgements: The study was supported by the project “Centre for Research of Patho-

genicity and Virulence of Parasites” (no. CZ.02.1.01/0.0/0.0/16_019/0000759) funded by

European Regional Development Fund and Ministry of Education, Youth and Sports of the

Czech Republic. Charles University institutional support (PROGRES Q43, UNCE 204017,

and SVV 244-260432/2017) applied to HD, LJ, and LM.

Abstracts

17

The effect of direct predation by fish on cercarial populations

of selected trematode species (Digenea)

P. Heinclová1, A. Born-Torrijos2, M. Soldánová2

1 Faculty of Science, University of South Bohemia in České Budějovice, Czechia



Recent studies suggest that feeding on parasites has important implications for parasite popu-

lation dynamics, disease risk control and food web structure in natural ecosystems. Parasites

with multi-host life cycles are usually trophically transmitted. However, parasites can also

function as a prey within ecosystems when predators actively consume their free-living stag-

es. The general energetic contribution of trematode cercariae, which are produced in high

quantities, to predator communities remains unknown for most aquatic systems. In this study,

we experimentally tested the ability of different freshwater non-host fish to consume cercariae

of selected trematode species with different species-specific traits (transmission mode, mor-

photype, size and behaviour). Two sets of experiments in a standard temperature 18°C were

designed using (i) a single dose of 100 cercariae, 4 trematode species (Cotylurus sp., Plagior-

chis elegans, Echinoparyphium aconiatum and Trichobilharzia szidati) and 4 fish predators

(Abramis bjoerkna, Oncorhynchus mykiss, Pseudorasbora parva and Rutilus rutilus); and (ii)

10 increasing doses of cercariae (2 to 800), 2 trematode species (P. elegans and T. szidati) and

1 fish predator of 2 different sizes (small and large P. parva). In all experiments, individually-

counted cercariae were offered to predators placed in containers with 500 ml water for 30

minutes. Results of the first experimental design showed that all predators were able to active-

ly feed on cercariae of all four species, significantly reducing their number in some predator-

prey combinations (35% of consumed cercariae on average, reaching up to 89%). The second

experiment showed that all fish-trematode combinations and groups exhibited a similar pat-

tern when the consumption rate increased with increasing parasites density (52% of predated

cercariae on average, reaching up to 73% at highest doses). The interspecific differences in

consumption patterns indicate the impact of species-specific traits with respect to the size,

swimming behaviour and speed of different cercariae species. Our results indicate that an im-

portant part of the cercarial biomass released from infected snail hosts to the environment

serves as food recourse for fish predators and thereby can have a strong effect on trematode

population dynamics, transmission and energy flow in aquatic food webs.

Acknowledgements: This study was supported by the Czech Science Foundation (Project No.

17-20936Y) and the Institute of Parasitology, Biology Centre of the Czech Academy of Sci-

ences (RVO: 60077344).

Abstracts

18

Entomopathogenic nematodes and insect immunity

P. Hyršl

Department of Animal Physiology and Immunology, Institute of Experimental Biology,

Masaryk University, Brno, Czechia

Entomopathogenic nematodes and their associated bacteria comprise together a highly patho-

genic complex which is able to invade and kill insect host within two days. Both bacteria and

nematodes produce a variety of factors interacting with the insect immune system and help to

overcome host defences. The tripartite model (Drosophila, nematodes, bacteria) was estab-

lished and used to show an immune function for candidate genes using different Drosophila

mutants or RNAi lines with defects in clotting or other branches of the immune system. Mi-

croarray analysis was used to compare gene expression of Drosophila larvae infected by the

entomopathogenic nematode Heterorhabditis bacteriophora and its symbiotic bacterium Pho-

torhabdus luminescens with non-infected larvae. The role of candidate genes, selected based

on genomic comparison, in response towards nematobacterial complex was further evaluated

by in vivo infection assays. We demonstrated an immune function during nematode infection

for known clotting enzymes and substrates, recognition molecules and eicosanoids. In conclu-

sion, we show that the Heterorhabditis/Photorhabdus infection model is suitable to identify

regulators of innate immunity in insects.

Acknowledgements: This study was supported by grant No. 17-03253S from the Czech Sci-

ence Foundation.

Abstracts

19

In silico design, construction and evaluation of multi-epitope chimeric antigens

for serodiagnosis of fasciolid flukes

M. Ježková1, R. Leontovyč1 and M. Kašný1,2

1 Department of Parasitology, Faculty of Science, Charles University, Prague, Czechia

2 Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czechia

Liver flukes of the family Fasciolidae are worldwide distributed serious pathogens of mam-

mals, including human. Two species occurring in the Czech Republic - Fasciola hepatica and

Fascioloides magna represent a serious risk to domestic and wild ruminants. The migrating

juveniles cause rare pathological changes in the body of definive hosts (small haemorrhagic

foci in the peritoneum layer). Blood feeding adults located in bile ducts (F. hepatica) or pseu-

docysts (F. magna) are responsible for severe damage of liver tissue (haemorrhages, necrotic

foci, chronic cholangitis, hyperplasia and fibrosis), which can lead to anemia, lose of weight

and even death of the host. Effective diagnostics is one of the key aspects of control of fasci-

olosis and fascioloidosis. Widely used immunological diagnostic methods, such as ELISA or

western-blot are often based on complex protein-antigens such as excretory-secretory prod-

ucts or whole single molecules e.g. cathepsins. This can complicate the standardisation of the

method and preparation of the recombinant form of protein serodiagnostic marker is time-

consuming. Nowadays the high throughput sequencing platforms provide the immense

amount of biological data, which can be beneficial for in silico antigen prediction. In present-

ed study we used Fascioloides magna transcriptomic and proteomic data for prediction of

antigenic epitopes using bioinformatic pipeline. In total, three different antigenic fusion pro-

teins, each comprises of three antigenic epitopes, were constructed and prepared in recombi-

nant forms. Their antigenicity and specificity were tested using ELISA and western-blot with

sera of infected animals (deer, goat sheep, rat, bull). Preliminary results showed that all de-

signed fusion proteins reacted with the sera from goats and bulls infected with F. hepatica.

Detection of F. magna infection in red deer and fallow deer has not been proved yet, but fur-

ther testing is in process. Here we present the complete workflow of in silico prediction, pro-

duction and testing of diagnostic protein-antigen markers for F. magna and F. hepatica. This

approach can be modified and applied for wide range of parasites.

Acknowledgements: Czech Science Foundation (No. 13-29577S)

Abstracts

20

Natural grazing: how to combine nature management,

big herbivores and their parasites

M. Jirků

Institute of Parasitology, Biology Centre of the Czech Academy of Sciences,


Česká krajina o. p. s./European Wildlife, Kutná Hora, Czechia

Natural grazing is relatively new concept of nature management by means of repatriation of

large herbivores. The first and still the only Czech grazing reserve is located in abandoned

military range Milovice near Prague. Essentials of the natural grazing concept, overview of

three years of experience and parasitological issues will be presented.

https://maps.google.com/?q=%C5%A0ultysova+170,+284+01+Kutn%C3%A1+Hora&entry=gmail&source=g

Abstracts

21

Failure to diverge connected with low rate of molecular evolution in an African

freshwater parasitic flatworm (Monogenea, Diplectanidae)

N. Kmentová1, M. Van Steenberge2,3, S. Koblmüller4, F. M. Bukinga5, T. M. N'sibula5,

P. M. Mulungula5, M. Gelnar1, M. P. M. Vanhove1,6,7


2 Biology Department, Royal Museum for Central Africa, Tervuren, Belgium 3 Operational Directorate Taxonomy and Phylogeny, Royal Belgian Institute of

Natural Sciences, Brussels, Belgium 4 Institute of Zoology, University of Graz, Graz, Austria

5 Centre de Recherche en Hydrobiologie, Département de Biologie, Uvira,

Democratic Republic of Congo 6 Hasselt University, Research Group Zoology: Biodiversity & Toxicology, Hasselt, Belgium

7 Finnish Museum of Natural History, Zoological Unit, University of Helsinki, Helsinki, Finland

Universal species identification using specific molecular tags derived from the cytochrome c

oxidase subunit 1 gene (COI) of the mitochondrial DNA, has successfully been implemented

in many taxonomic groups. Correspondingly, molecular characterisation has revealed cryptic

species. However, incongruence between morphological and genetic results in well-studied

monogenean species was documented in members of Diplectanidae (Monogenea). In our

study, we tested the link between the level of genetic and morphological differentiation for

diplectanid parasites infecting latid hosts in geographically isolated areas in Lake Albert and

Lake Tanganyika.

Specimens from five African latid species (Lates angustifrons, L. mariae, L. microlepis, L.

niloticus, L. stappersii) were examined for the presence of monogenean gill parasites from

several localities. Monogeneans were characterised morphologically via morphometrics of

sclerotised structures and genetically using markers with different rates of molecular evolu-

tion from nuclear ribosomal and mitochondrial DNA regions.

In total, four out of five latid species were infected by monogenean parasites belonging to

Diplectanum. Continuous morphological variation was documented, and the genetic distance

between parasites of geographically isolated host species in the COI region did not reach the

previously published threshold of 14.5%. Therefore, a single species, D. lacustre, is suggested

to infect latids in both lakes. Based on the age of the eastern African Rift Lake system (9

MYA), the substitution rate in the COI region of D. lacustre was estimated to be 0.5%. De-

spite the persistent geographic barrier, D. lacustre has not reached the threshold of genetic

differentiation in COI gene proposed for species delineation of diplectanid monogeneans.

Congruence between the slow rate of molecular evolution in the mitochondrial COI region

and the level of parasite morphological differentiation is suggested.

Acknowledgements: This study was supported by ECIP (European Centre of IchthyoParasit-

ology); centre of excellence program of the Czech Science Foundation; project No.

P505/12/G112.

Abstracts

22

Alveolar echinococcosis in the Czech Republic – omnipresent?

L. Kolářová

Institute of Immunology and Microbiology, 1st Faculty of Medicine, Charles University and

National Reference Laboratory for Tissue Helminthoses, General University Hospital in

Prague, Czechia

Alveolar echinococcosis (AE) represents a zoonotic infection characterized by faecal–oral route

of transmission. It is caused by metacestodes of Echinococcus multilocularis and can affect

both humans and animals. Usually, the host liver are affected, however the parasites inducing

tumor–like lesions can proliferate to adjacent as well as to distant organs. The infection is

acquired after ingestion of the parasite eggs produced by adult cestodes living in the intestine of

various carnivores. In the Czech Republic, screening examinations for detecting specific

infectious agents are performed since 1994. The first case of human AE has been detected in

2007. According to radiological, histological and/or PCR data, serological examinations of

2,958 individuals helped to reveal 41 AE cases till present. Whole data from 2007 - 2017

demonstrated AE in 21 men and 20 women. The age of onset varied from 7 to 82 years old and

was lower in women (mean 43.7, median 39.5) than in men (mean 53.6, median 58). To exam-

ine differences in non–Gaussian distributed variables between males and females, the age data

were analyzed using 2–way Mann–Whitney non–parametric test which showed no statistically

significant differences in age between males and females when P>0.1468. Our study indicates

an increase in the number of persons with AE. Anamnestical data of such patients confirmed

autochtonous character of the infections in the CR. This can be also documented by findings of

the metacestodes in wild boar, bank vole, golden retriever and zoo animals as well as of the

adult cestodes in red foxes, dogs, cats and raccoon dog which were published by several

authors. According to literary data, the mean prevalence rate of E. multilocularis ranges

between 14 – 62 % (mean 33 %) in foxes in our country. In the context of known data on

detection of various parasite developmental stages in humans and animals from various parts of

the country, AE infection risk seems to be omnipresents in the CR.

Acknowledgement: The study was supported by the Charles University (Research Programs

PROGRES No. Q25). Our thanks also go to Markéta Leissová for technical assistance during

the examination.

Abstracts

23

Characterisation of selected peptidases from Trichobilharzia regenti cercariae

L. Konečný1, M. Kašný1,2, L. Mikeš1, P. Horák1

1 Department of Parasitology, Faculty of Science, Charles University in Prague, Czechia.

2 Department of Botany and Zoology, Faculty of Science, Masaryk University Brno, Czechia.

Trichobilharzia regenti is a neurotropic parasite of birds from the same family Schistoso-

matidae like the notoriously known blood-dwelling fluke Schistosoma mansoni, which cause

serious human disease - schistosomiasis. Cercariae, the invasive stages of these trematodes

actively penetrate the hosts skin employing excretory-secretory products (ESP), which contain

proteolytic enzymes able to disrupt host tissues and thus reach the successful transmission.

Since 1950s, when the first experiments with cercarial extracts were performed, various tech-

niques for collection of ESP have been tested. The first aim of our work is to compare the

protein composition of T. regenti cercarial ESP obtained after two stimulation methods - by

linoleic acid and duck skin.

The most abundant secreted enzyme responsible for cercarial penetration of human schisto-

some S. mansoni is cercarial elastase. This serine protease is well known to degrade skin pro-

teins such as elastin, keratin, collagens, fibronectins and laminin. On the other hand, bird

schistosomes from genus Trichobilharzia use cysteine-peptidases, particularly cathepsins.

Recently, the gene coding the cercarial elastase in T. regenti genome was identified. Our se-

cond aim is to produce the recombinant form of T. regenti cercarial elastase, characterize its

biochemical properties and localize this protein in different ontogenetic stages of the parasite.

Abstracts

24

Impact of the fish hybridization on the parasite community:

specificity, diversity and co-evolution

V. Krasnovyd, A. Šimková

Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czechia

Influence of the host hybridization and its consequences on the parasite community formation

of the fish hybrids have been in focus for past few decades. Limited mainly by the temporal

variability, the poikilothermic fish host is hybridizing frequently in nature. The hybridization

in fish influences their physiology, immunity and ecology. The interspecific hybrids represent

the models to study genetic limits of host specificity of parasite species, host resistance/sus-

ceptibility and co-evolutionary host-parasite relationships. Role of the fish hybridization and

maternal origin of hybrids on the parasite diversity and specifically on the host-specific para-

sites were studied. The level of parasite infection in two hybridizing cyprinid systems was

studied. The first system studied represents the natural hybridization between evolutionary

divergent species - roach (Rutilus rutilus) and common bream (Abramis brama). In this sys-

tem, morphological traits (anal fin ray number, number of gill rakers, number of scales in lat-

eral line) and genetic markers (microsatellites; cyt b partial gene) were used for individual

determination. The second system studied represents the artificially prepared pure and F1 hy-

brid lines of two phylogenetically related species - the silver bream (Blicca bjoerkna) and

common bream (A. brama). The analyses using both systems indicate the higher parasite spe-

cies richness in F1 hybrids when compared to their parental species. Specialist and generalist

parasite species of the parental fish were present in the parasite communities of the cyprinid

hybrids. However, several parasite species exhibiting the strict host specificity were not pre-

sent in the parasite communities of hybrids. It may be related to the different level of the host-

parasite co-adaptation between host-specific parasite species and its host species. The parasite

community composition might be dependent on the degree of the genetic introgression be-

tween hybridizing host species and parasite affinity to the specific fish host.

Acknowledgements: This study was funded by Czech Science Foundation, project no.

P505/12/0375. Vadym Krasnovyd was funded by Czech Science Foundation, ECIP - Centre of

Excellence, project no. P505/12/G112.

Abstracts

25

Asian fish tapeworm: the most successful invasive parasite in freshwaters

R. Kuchta1, A. Choudhury2, T. Scholz1


České Budějovice, Czechia 2 Division of Natural Sciences, St Norbert College, De Pere, Wisconsin, USA

The Asian fish tapeworm (AFT), Schyzocotyle acheilognathi, is a notorious and highly suc-

cessful invasive parasite reported in a wide spectrum of freshwater fishes, and new reports of

its spread continue to emerge. To date, no thorough review of its worldwide distribution and

host associations is available. In the present work, we collected information from 651 articles

up until 2017, from which we updated the number of the hosts to 312 fish species and 11 non-

fish species, which is quite unusual among helminths. The AFT has spread to all but one con-

tinent (Antarctica). The highest number of records are from North America, followed by Asia

and Europe. A key feature of its invasive success is its broad environmental tolerance.

Acknowledgements: This study was financially supported by the Czech Science Foundation

(15-14198S) and the Institute of Parasitology, BC CAS (60077344).

Abstracts

26

Schistosomes and their smart weapons – proteases

A. Leontovyč1, 2, J. Dvořák1,3

1 Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Prague, Czechia

2 First Faculty of Medicine, Charles University, Prague, Czechia 3 Department of Zoology and Fisheries, Faculty of Agrobiology, Food and Natural Resources,

Czech University of Life Sciences Prague, Czechia

Schistosomiasis caused by parasitic blood flukes of the genus Schistosoma is the second most

important parasitic infection after malaria with 250 million people infected worldwide and

many more at risk. Therapy heavily relies on a single drug – praziquantel, hence there is an

urgent need to identify novel anti-schistosomal targets for therapeutic interventions; and pro-

teolytic enzymes of schistosomes are attractive drug targets.

Adult schistosomes reside in blood stream of the final host and feed on blood. Hemoglobin is

the major source of nutrients necessary for the growth, development and reproduction of the

parasite. Its digestion is mediated by the proteolytic arsenal of the schistosome digestive tract,

which includes enzymes with complementary specificity belonging to the classes of cysteine

and aspartic proteases, and metalloproteases. But proteolysis of host proteins is not limited

just to the gut. There are a number of proteases secreted by the tegument (surface of the

worm), which play crucial roles in various processes of the host, such as vasodilatation, blood

coagulation or immune system evasion. These enzymes are most likely able to interfere with

the host regulatory pathways and provide survival benefit to the schistosomes.

The proteolytic system is essential for the viability of schistosomes and is a current topic of

intense research focused on the development of new vaccines and chemotherapeutics for the

treatment of schistosomiasis.

Abstracts

27

Comparative transcriptomic analysis shows molecular mechanisms used

by visceral and neurotropic avian schistosomes

R. Leontovyč1, N. Young2, M. Kašný1,3, R. Gasser2, P. Horák1


2 Faculty of Veterinary Science, The University of Melbourne, Australia 3 Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czechia

Avian schistosomes of the genus Trichobilharzia are in focus of parasitologists due to medi-

cal importance of cercarial dermatitis, they cause to humans. In addition, T. regenti induce the

neuropathogeny in its permissive avian hosts and accidental mammalian hosts. To date main-

ly human pathogens of the genus Schistosoma are intensively studied, and the other schisto-

somatids are somewhat neglected in terms of molecular investigations. Trichobilharzia szidati

and T. regenti belonging to the same genus differ in their life strategy in definitive hosts. Sim-

ilarly to human schistosomes, T. szidati schistosomula migrate via circulatory system to the

lungs, liver and visceral veins. On the other hand, T. regenti undergoes a unique neurotropic

migratory route via peripheral nerves, the spinal cord, and brain up to the nasal mucosa. Using

light microscopy and transmission electron microscopy, the lung schistosomula (T. szidati)

and schistosomula in the spinal cord (T. regenti) have been explored for their localization in

the tissues, interaction with the host and nutrition preference. Results have been confronted

with transcriptomic data of two consecutive stages, cercariae and schistosomula of both spe-

cies, to elucidate the spectra of synthesized proteins in the early and late phases of infection.

Despite the same mission of cercariae of both species, i.e., to find and attack the definitive

host, level of transcription profiles differ in, e.g., ribosomal proteins, and components of en-

ergy and lipid metabolisms. Differences between schistosomulum stages have been observed

in, e.g., signaling, transport system and regulation of transcription. Also, the detailed expres-

sion profiles of peptidases have been compared and linked to different nutrition preferences of

both species. Our results provide new insights into the molecular biology of avian schisto-

somes and can be used as a comparative platform for the research of schistosomatids and oth-

er trematodes.

Acknowledgements: The study was supported by Czech Science Foundation (Grant Nos. 13-

29577S and 18-11140S) and by the project “Centre for Research of Pathogenicity and Viru-

lence of Parasites” (no. CZ.02.1.01/0.0/0.0/16_019/0000759) funded by European Regional

Development Fund and Ministry of Education, Youth and Sports of the Czech Republic.

Charles University institutional support (PROGRES Q43, UNCE 204017, and SVV 244-

260432/2017)

Abstracts

28

A new insight into the classification of the Ascaridoidea

L. Li1,2, L. Lü1, S. A. Nadler3, D. I. Gibson4, L.-P. Zhang1

1College of Life Sciences, Hebei Normal University, Shijiazhuang, P. R. China


České Budějovice, Czechia 3Department of Entomology and Nematology, University of California, Davis, USA

4Department of Life Sciences, Natural History Museum, London, UK

Ascaridoids are among the commonest groups of zooparasitic nematodes (roundworms) and

occur in the alimentary canal of all major vertebrate groups, including humans. They have an

extremely high diversity and are of major socio-economic importance. Current classifications

of the Ascaridoidea have been proposed based mainly on morphological characters, including

the form of the excretory system, the structure of the oesophago-intestinal junction, the labial

structure and the number and arrangement of caudal papillae. The different weighting of these

morphological characters by individual investigators has led to conflicting classifications of

this superfamily. In this study, we performed phylogenetic analyses of ascaridoid nematodes

based on five nuclear [partial sequences of 18S, 28S, ITS-1, 5.8S and ITS-2] and three mito-

chondrial genes [partial cox1, cox2 and 12S]. These molecular phylogenetic analyses included

the most comprehensive taxon sampling of the Ascaridoidea to date. The results indicated that

the representatives of the Ascaridoidea were divided into six distinct clades (families): Het-

erocheilidae, Anisakidae, Toxocaridae, Acanthocheilidae, Ascarididae and Raphidascarididae,

with strong support from both maximum likelihood (BS = 99-100) and Bayesian inference

(BPP = 1), except for the Toxocaridae (BPP = 0.98, BS = 58). These analyses further confirm

the Heterocheilidae as the sister clade to the remaining Ascaridoidea. Our present study repre-

sents the first attempt to resolve the systematic position of the Acanthocheilidae using phylo-

genetic analyses based on molecular data. The results support the validity of the Acanthochei-

lidae as a distinct family outside the Anisakidae, but these results strongly reject these pro-

posals to remove Pseudanisakis from the Acanthocheilidae. Our phylogenetic analyses also

confirmed Mawsonascaris as a member of the Acanthocheilidae with a sister relationship to

Acanthocheilus + Pseudanisakis.

Acknowledgements: This study was supported by the National Natural Science Foundation of

China (NSFC-31572231, NSFC-31750002).

Abstracts

29

Dynamics of immune cells in the CNS of mice after infection by Trichobilharzia regenti

T. Macháček1, K. Immig2, P. Horák1


2 Institute of Anatomy, Faculty of Medicine, Leipzig University, Leipzig, Germany

The neuropathogenic schistosome Trichobilharzia regenti infects the central nervous system

(CNS) of vertebrates, such as ducks or mice. In both hosts, the affected nervous tissue gets

inflamed and infiltrated by peripheral leukocytes. However, the detailed progression of this

process, including determination and quantification of the immune cells, is not known. In this

study, we applied flow cytometry (FC) to characterize the dynamics of immune cells present

in the CNS of C57BL/6J mice following T. regenti infection. First, the immune cells from the

spinal cords and the brains of the mice were isolated by Percoll gradient 0, 7, 14, 21, and 28

days post infection (dpi). Granulocytes, T-lymphocytes, and microglia were then identified by

FC according to cell granularity and expression of the surface markers (CD45, C11b, CD115,

and CD3). The influx of granulocytes and T-lymphocytes into the spinal cord peaked 14 dpi.

Microglia also expanded throughout the infection being the major MHC II+ cell population.

Unexpectedly, granulocytes and T-lymphocytes infiltrated also the brain, mainly 21 dpi, alt-

hough the tissue was mostly free from parasite DNA as revealed by qPCR. Contrary to the

spinal cord, expression of MHC II was inconspicuous on brain microglia, suggesting rather “a

resting state”. Taken together, these results present the first quantitative look at the dynamics

of immune cells in the CNS of mice infected by T. regenti. We observed activation and ex-

pansion of microglia during the infection, and we also identified granulocytes as the major

cell population infiltrating the CNS.

Acknowledgements: This study was supported by Czech Science Foundation (13-29577S, 18-

11140S).

Abstracts

30

Does the neurotropic helminth influence the peripheral immune response?

M. Majer, T. Macháček, P. Horák


The neurotropic avian schistosome Trichobilharzia regenti can accidentally infect mammals

including mice. However, the parasite dies during its migration through the host skin (within

1 week) or the central nervous system (CNS) (within next 2–3 weeks). Although the inflam-

matory reaction develops in the affected nervous tissue, almost nothing is known about the

effect of infection on the peripheral immune response. In the present study, we used flow cy-

tometry to characterize CD3+ CD4+ T cells isolated from the spleen of mice 0, 3, 7, 14, and

21 days post infection (dpi) by T. regenti. Different T cell subsets were distinguished by their

expression of intracellular cytokines and transcription factors: pro-inflammatory Th1 (IFNγ,

T-bet) and Th17 (IL-17, RORγt), anti-inflammatory Th2 (IL-4, GATA-3), and regulatory

Treg (IL-10, FOXP3). Preliminary data indicate an unexpected decrease of all splenic T cell

subsets, except for Treg cells, 3 dpi when most of the parasites are trapped and killed by the

immune system in the skin and the rest of them appear intact in the spinal cord. The amount

of splenic Th1, but not Th17 cells, grows rapidly 7 dpi when the inflammatory reaction

around the parasite in the spinal cord is observed. Most of the parasites present in the spinal

cord are usually found damaged and dead 14 and 21 dpi, when we noticed an increased

amount of splenocytes producing IL-4 (possibly Th2 cells) and Treg cells, respectively. Dif-

ferentiation of these T cell subsets in the late phase of infection suggests a linkage between

the immune response in the CNS and the periphery. On the other hand, the Th1 response at

the beginning of infection could be explained not only by the developing inflammation in the

CNS but also by the inflammation in the skin, which shows a mixed Th1/Th2 polarization, as

described previously.

Acknowledgement: This work was supported by Czech Science Foundation (18-11140S).

Abstracts

31

The localization of glycogen in the mature spermatozoa

of some caryophyllidean tapeworms

M. Matoušková1, M. Bruňanská1, M. Oros1, J. Nebesářová2

1 Institute of Parasitology, SAS, Košice, Slovak Republic

2 Institute of Parasitology, Biology Centre CAS, České Budějovice, Czechia

Male gametes of three caryophyllidean tapeworms (Khawia rossittensis, Caryophyllaeides

fennica and Archigetes sieboldi) have been examined using the periodic acid-thiosemicarb-

azide-silver proteinate (PATSC-SP) technique of Thiéry (1967) for glycogen. Adult parasites

were isolated from intestines of freshwater cyprinid fishes snared in Tisa River, East Slovakia.

Tissue samples were fixed in cold 2.5% or 3 % glutaraldehyde in sodium cacodylate buffer

(pH 7.4), and prepared routinely for TEM observations. Ultrathin sections were cut using

Leica Ultracut UCT ultramicrotome, placed on gold grids, treated by solution of periodic acid,

rinsed and washed in destilled water, followed by application of thiosemicarbazide, washing

in acetic acid, rinsing with destilled water, application of silver proteinate, and final washing

in some portions of destilled water. The presence of glycogen was detected in the cytoplasm

of principal regions (II, III, and IV) of the mature spermatozoa of caryophyllideans studied. In

contrast to some other flatworms having male gametes with the 9 + “1” trepaxonematan struc-

ture, intraaxonemal glycogen was not observed in the mature spermatozoa of caryophyllidean

tapeworms.

Acknowledgements: This study was supported by the Grant Agency of the Slovak Republic

VEGA (project no. 2/0104/16 to MB). We acknowledge the core facility of the Institute of

Parasitology, BC ASCR in České Budějovice, Czech Republic, supported by the MEYS CR

(LM2015062 Czech-BioImaging). This research was undertaken within the framework of a

joint research projects Nos. AV ČR-16-08 and SAV-18-21 supported by a bilateral agreement

on scientific exchange and cooperation signed by the Czech and Slovak Academies of Scienc-

es. The work was realized within a frame of Centre of Excellence for Parasitology (Code

ITMS: 26220120022) based on the support of the Operational Programme “Research & De-

velopment” funded from the European regional Development Fund (rate 0.4). Financial sup-

port of The National Scholarship Programme provided by The Ministry of Education, Sci-

ence, Research and Sport of the Slovak Republic is gratefully acknowledged.

Abstracts

32

Worms as relevant biological models for investigation of Glutamate carboxypeptidase 2

D. Matušková1, T. Šulcová1, E. M. Boateng1, L. Ulrychová 2,3, M. Asahina4,

C. Bařinka5, J. Dvořák1,3

1 Department of Zoology and Fisheries, Faculty of Agrobiology, Food and Natural Resources,

Czech University of Life Sciences Prague, Czechia 2 Department of Parasitology, Faculty of Science, Charles University, Prague, Czechia

3 Institute of Organic Chemistry and Biochemistry, The Czech Academy of Sciences,

Prague, Czechia 4 Department of Cellular and Molecular Pharmacology, University of California,

San Francisco, USA 5 Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Vestec, Czechia

Glutamate carboxypeptidase 2 (GCP2) is an established cancer marker and target for therapies

of neurological disorders, but its physiological roles in humans are not fully understood. Less-

complex organisms offer attractive tools to study basic functions of GCP2, as its orthologs are

found in all living phyla. The proposal aims at detailed characterization of GCP2 in two mo-

del organisms: platyhelmints (Schistosoma mansoni) and nematodes (Caenorhabditis ele-

gans). We will express and purify their GCP2 orthologs and profile their substrate specificity

using combinatorial libraries, complemented by structural and molecular modeling studies.

Biological experiments will focus on expression profiling (RNA and protein level, transgenic

technologies) and localizations in developmental stages and individual cells, followed by tar-

geted approaches to pinpoint GCP2 importance for physiology and survival of studied worms.

Combined, understanding of ancestral roles of GCP2 in non-mammalian species can guide

future studies in other species and offer a starting point for drug/vaccine development against

some groups of helminths in future.

Abstracts

33

Digeneans of the family Strigeidae Railliet, 1919 from Iceland:

morphology and molecular characterisation

H. Mazanec1,2, J. Roháčová1,2, S. Georgieva2, K. Skírnisson3, A. Faltýnková2

1 University of South Bohemia in České Budějovice, Faculty of Science, České Budějovice, Czechia


České Budějovice, Czechia 3 Laboratory of Parasitology, Institute for Experimental Pathology, Keldur,

University of Iceland, Reykjavík, Iceland

Iceland, a volcanic island in the sub-Arctic, is characteristic for its depauperate freshwater and

terrestrial fauna. Thus, as all other areas at high latitudes, its ecosystems are considered to be

simple, of low diversity, with short trophic linkages, few pathogens and a limited capacity for

adaptation to environmental change. However, recent molecular studies have revealed consid-

erable unrecognised species diversity within trematodes not only in Iceland (Diplostomum

spp.), but also in Northern Europe and Northern America. Our understanding of the diversity

of northern parasites, host-parasite associations and the detection of disease emergence de-

pends strongly on accurate parasite identification. Therefore, the application of comparative

morphological and molecular approaches to species delimitation is the best approach. Totals

of 11 bird species, three fish and three snail species were sampled and examined for the pres-

ence of trematodes in Iceland in 2009–2017. In the present study, 90 isolates of representa-

tives of the family Strigeidae Railliet, 1919 were subjected to morphological and molecular

assessment. Partial sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and

nuclear 28S rRNA genes were used for molecular identification, and corresponding life-cycle

stages were matched. Integrated morphological and molecular analyses demonstrated the

presence of seven species/lineages in Iceland represented by larval and/or adult isolates: two

species of Apatemon Szidat, 1928; three species of Australapatemon Sudarikov, 1959, and

two lineages of Cotylurus Szidat, 1928. The life-cycle was fully resolved for Apatemon gra-

cilis (Rudolphi, 1819). The life-cycles of Australapatemon burti (Miller, 1923) and A. minor

(Yamaguti, 1933) were resolved in part only, because leeches (the second intermediate hosts)

were not yet examined. Furthermore, we obtained data on two Cotylurus lineages, one from

birds (adults) and the other from snails (cercariae and metacercariae). Based on the collected

data we corroborate the snail Radix balthica as the main intermediate host for trematodes in

sub-Arctic Europe, and migratory birds (Anatidae and Gaviidae) as the main factors maintain-

ing populations of strigeid trematodes in Iceland. However, to prove if the trematode life-

cycles are being fully completed in Iceland, further analyses of the present data and more ma-

terial are necessary. This is the first molecular confirmation of species identity of strigeid

trematodes in Iceland.

Acknowledgements: This study was supported by the Research Fund of the University of Ice-

land, the Czech Science Foundation (18-18597S) and by the Student Grant Agency of the

Faculty of Science, University of South Bohemia in České Budějovice (G103204RVOKPA).

Abstracts

34

Cysteine and aspartic peptidases in the blood-feeding monogenean

Eudiplozoon nipponicum

L. Jedličková1, H. Dvořáková1, J. Dvořák2, L. Ulrychová3, V. Žárský1, M. Kašný1,4, L. Mikeš1

1 Department of Parasitology, Faculty of Science, Charles University, Prague 2, Czechia

2 Department of Zoology and Fisheries, Faculty of Agrobiology, Food and Natural Resources,

Czech University of Life Sciences Prague, Czechia 3 Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,

Czechia 4 Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czechia

Analysis of Eudiplozoon nipponicum transcriptome revealed numerous transcripts of cysteine

and aspartic peptidases – namely a set of 10 cathepsins L (EnCL), 1 cathepsin B (EnCB) and 3

cathepsins D (EnCD). A structural analysis of EnCLs had shown significant variability in their

active sites, specifically the S2 subsites responsible for interaction with substrates. Surprisingly,

we found only one E. nipponicum cathepsin B (EnCB) transcript which showed similarity with

cathepsin B2 of Schistosoma mansoni. Two of the three EnCDs possess complete active sites,

while the third one is a paralog with a mutated active site Asp. Two genes encoding abundant

transcripts of EnCLs, the gene encoding EnCB, and genes encoding the two functional EnCDs

were cloned and functionally expressed in yeast or bacteria. All recombinant cathepsins were

able to hydrolyze blood proteins or collagen I. Monospecific antibodies and specific RNA

probes localised the EnCL and EnCD2 proteins/transcripts to the haematin cells of the worm’s

digestive tract and gut lumen. EnCB displays molecular features typical of cathepsins B, includ-

ing an occluding loop responsible for its exopeptidase activity. Although the EnCB hydrolysed

haemoglobin in vitro, it was localised in the vitelline cells of the parasite and not the digestive

tract. This study represents the first bioinformatic analysis and biochemical characterisation of

cysteine and aspartic peptidases in the monogeneans. E. nipponicum expresses a variety of ca-

thepsins L, which are the most abundant peptidases in the adult worms. The properties and lo-

calisation of EnCLs and EnCD2 suggest a central role in the (partially extracellular?) digestion

of host blood proteins. High variability of substrate-binding sites in EnCL enzymes suggests

specific adaptation to a range of biological processes that require proteolysis. A single cathepsin

B expressed by the parasite probably plays a role in vitellogenesis. Therefore, it seems that ca-

thepsins L and D, but not cathepsin B, contribute to the complex intestinal proteolytic network

in the blood-feeding monogenean E. nipponicum.

Acknowledgements: Centre for Research of Pathogenicity and Virulence of Parasites” (no.

CZ.02.1.01/0.0/0.0/16_019/0000759) funded by European Regional Development Fund and

Ministry of Education, Youth and Sports of the Czech Republic. Czech Science Foundation

grants (P506/12/1258, GBP505/12/G112), Charles University institutional support (PRO-

GRES Q43, UNCE 204017, and SVV 244-260432/2017), Masaryk University (MU-

NI/A/1362/2016), Ministry of Education, Youth, and Sports of the Czech Republic (InterBio-

Med LO1302), and by the IOCB institutional project (RVO 61388963).

Abstracts

35

Amphibians and their helminth parasites: macroecological patterns and knowledge gaps

S. Morand

Centre national de la recherche scientifique, Institut des Sciences de l’Evolution de Mont-

pellier, Université Montpellier - CIRAD ASTRE, France

Faculty of Veterinary Technology, Kasetsart University, Bangkok, Thailand

While the knowledge on the diversity, including genetic diversity, and distribution of amphi-

bians have improved over the last decades, the knowledge on the diversity of their parasites is

particularly scarce, specially in Southeast Asia (a hotspot of biodiversity at threat). Despite

the threats of global emergence of infectious diseases to amphibian species worldwide, we

still do not know some of the basic elements on their parasites, and diseases, and how these

parasites may impact the life-histories of their amphibian hosts. Here, I will first review the

knowledge on the diversity and distribution of amphibians, with a focus on Southeast Asia.

Then, I will use several databases to synthesize the knowledge on the diversity and distributi-

on of their helminth parasites. Combining both data on parasites and their amphibian hosts

and using a macroecological framework of host-parasite interactions will help identify gaps in

knowledge of amphibians-parasites interactions. Finally, I will show why, and how, studying

parasites of amphibians is a key point for studies in conservation biology of amphibians or in

health ecology by emphasizing life-history traits and immuno-ecology.

Abstracts

36

Occurrence of the zoonotic parasites Anisakis spp. in Atlantic herring (Clupea harengus)

marketed in Slovakia

M. Oros, D. Barčák, D. Uhrovič

Institute of Parasitology, Slovak Academy of Sciences, Košice, Slovakia

Anisakiasis is an emerging zoonosis caused by the fish parasitic nematode larvae belonging to

the genus Anisakis (Nematoda, Anisakidae), parasites of marine and anadromous fishes. Dur-

ing the years 2017 – 2018, a total of 100 whole individuals of Atlantic herring (Clupea ha-

rengus), marketed to the fish processing facility, and 18 specimens of pickled herrings, ready

to eat product from the food trades in Slovakia, were examined for the presence of Anisakis

spp. The parasite location and degree of infestation were evaluated by using direct observa-

tion and UV diaphanoscopy. The coelomic cavity, gonads (testes and ovary) and muscles

have been carefully examined in all individual fish. A total of 4163 Anisakis third stage larvae

(L3) were collected from this commercially important food fish species, with the overall

prevalence 100% and a mean intensity of 42 (2–368). The majority of Anisakis larvae, i. e.

4018 individuals, were detected in the abdominal cavity (prevalence 100%), 74 in the gonads

(21%), and 71 were found in the fish flesh (33%). Anisakis larvae were also found in 6 pick-

led herrings ready to eat products and 21 larvae were collected, with a range 1–9 per fish

product. Anisakidae larvae found in Atlantic herrings were randomly chosen for

morphological characterization based on the length and shape of the ventriculus, the cephalic

head and the caudal tail appearance. The genetic identification was performed using BLAST

algorithm comparing ITS1-5.8S-ITS2 rDNA sequence region with data in GenBank. Our pre-

liminary data showed high occurrence of the Anisakis simplex, which have the highest poten-

tial to cause anisakiasis. Present investigation showed that commercially important food fish

species heavily infected with Anisakis spp. larvae reach the fish processing industry, and sub-

sequently, ready to eat fish products contaminated with Anisakis reach the market.

This study was supported by the SAS-MOST Joint Research Project (no. 2016/7).

Abstracts

37

In vivo imaging techniques for parasitology research

J. Pankrác1, B. Vojtková2, V. Kolářová1, J. Sádlová2, L. Šefc1

1 Center for Advanced Preclinical Imaging, First Faculty of Medicine,

Charles University, Prague, Czechia 2 Department of Parasitology, Faculty of Science, Charles University, Prague, Czechia

Non-invasive in vivo imaging methods allow detailed anatomical imaging (X-ray, computed

tomography, ultrasonography, magnetic resonance imaging), and molecular imaging (positron

emission tomography, single-photon emission computed tomography, magnetic particle imag-

ing, photoacoustic imaging, and optical imaging of fluorescence, bioluminescence and Che-

renkov radiation). Most of these techniques were proved to be essential for human diagnostics

including diagnostics of parasitic infections and accompanying pathologic changes. The main

advantage of using modern in vivo imaging methods is the possibility to record precisely the

morphology but also dynamic changes in metabolic processes and distribution of molecules of

interest without harming the patient. Nowadays, high spatial resolution of non-invasive imag-

ing methods facilitates working also with small animal models like mouse, rat or fish. This

allows to carry out within-subject design where the animal serves as its own control and high

statistical power is achieved even with low number of animals. Utilizing these tools in the

helminthology would enable tracking of migration routes, monitoring the changes in parasite

burden and immune response, localization of cysts or visualization of morphological changes

in organs. Currently, in vivo imaging methods are still rarely used in basic helminthology re-

search. In this presentation, we survey the imaging methods currently used to visualize hel-

minths in small animal models – positron emission tomography (PET) and optical imaging.

Ultrasonography and photoacoustics are also included as methods with high potential for

small animal imaging.

Acknowledgements: The Ministry of Education, Youth and Sports (LM2015062-

CzechBioImaging and SVV 260371/2018), Czech Science Foundation GACR (grant number

17-01911S) and GA UK (grant number 288217).

Abstracts

38

Fascioloides magna digestive peptidases - cathepsin L, B and D:

gene expression and tissue localization of their transcripts

K. Peterková1, P. Horák1, M. Kašný1,2

1 Department of Parasitology, Faculty of Science, Charles University in Prague, Czechia.

2 Department of Botany and Zoology, Faculty of Science, Masaryk University Brno, Czechia.

Fascioloides magna (also known as a giant liver fluke) is an important parasite causing fasci-

oloidosis of wild and domestic ruminants in Europe and North America. This digenetic trema-

tode feeds on host blood and our previous research showed that one of proteolytic enzymes -

cathepsin L (FmCL, cysteine peptidase) is predominantly responsible for the blood digestion.

The transcripts of FmCL were indentified also in other ontogenetic stages which do not up-

take any nutrients, suggesting the possible different function of the enzyme. The aim of our

work is to evaluate the FmCL expression level using qPCR, localize its expression within

bodies of adult worms, eggs and miracidia using the RNA in situ hybridization and compare

the results to other F. magna cysteine peptidases – cathepsin B (FmCB) and D (FmCD). We

revealed that FmCL is approximately 1500 times more expressed by adults than by eggs and

miracidia. We confirmed, that FmCL is intensively transcribed in the cells of adults´ gastro-

dermis and less in the organs of reproductive system such as vitellaria, ovary, Mehlis glands,

uterus and testes. Similar results were obtained with FmCB and FmCD. These findings sup-

port our hypothesis that cathepsin L is probably the major digestive peptidase of F. magna,

but it also may have other biological functions.

Abstracts

39

The past and the present of avian schistosomes and cercarial dermatitis in the Czech

Republic

Z. Pokrupová, J. Bulantová, P. Horák


Avian schistosomes (Trematoda, Schistosomatidae) can primarily be found in birds connected

with water environment, mainly in the order Anseriformes. Larvae of these flukes – cercariae

– develop in fresh or marine water snails. After their maturation, cercariae are shed to the wa-

ter environment. When they find an appropriate avian host, they penetrate its skin and develop

to adults. They can also accidentally penetrate the human skin. Here, the cercariae cannot

complete their life cycle but they can cause an allergic skin reaction called cercarial dermatitis

(CD) which is the main reason for studying these parasites. The first record of CD in the

world was described in 1928 from the Michigan lake in the USA, and the records from other

localities including European countries followed; first European cases were described in 1930

from the UK, Germany and France. The condition occurred probably earlier but without de-

termination of cercariae as the causative agent. The history of CD in the Czech Republic is

dated back to 1966 when it was caused by the genus Trichobilharzia (T. ocellata/T. szidati),

one of the most often genera associated with CD. During the years, more species and genera

of avian schistosomes were recorded in the Czech Republic (T. regenti, T. franki, T. filiformis,

Bilharziella polonica, Gigantobilharzia vittensis, G. acotylea, Dendritobilharzia pulverulenta

or Ornithobilharzia canaliculata) but not all of them were proven to be able to cause CD in

humans. During the field studies, it is not always possible to determine the causative agent of

CD at the species level, eventually identify the intermediate or definitive hosts. Also, histori-

cal findings are usually limited only to morphological characterization of cercariae from the

snails, and adult worms from the dissected birds. Nowadays, CD is a problem mainly in natu-

ral recreational water bodies where it can cause economic losses. Together with increasing

importance of CD, the older morphological data are now correlated with the current methods

of molecular biology. Despite this fact, a summarizing list of the occurrence of CD or avian

schistosomes in the Czech Republic is still unavailable. Hence, we decided to find any availa-

ble information in scientific publications, MSc. or PhD. theses, reports from regional hygiene

stations and articles from local newspapers, and to create a map of the Czech Republic with

gained information included which will be available to the public.

Acknowledgements: Czech Science Foundation (Grant No. 18-11140S)

Abstracts

40

Association between pharmaceuticals and parasite infection

in natural brown trout (Salmo trutta) populations

M. Pravdová1,2, M. Ondračková1, J. Kolářová3, K. Grabicová3, T. Randák3, P. Jurajda1

1 The Czech Academy of Sciences, Institute of Vertebrate Biology, Brno, Czechia

2 Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czechia 3 University of South Bohemia in České Budejovice, Faculty of Fisheries and Protection of

Waters, Research Institute of Fish Culture and Hydrobiology, Vodňany, Czechia

Contaminants in the natural environment are known to affect aquatic biota. In recent years,

the role of pharmaceutical contaminants has grown in importance due to the ever increasing

amounts released into the environment and their biological activity. Sewage treatment plants

represent one of the most important sources of such contaminants. In addition to direct nega-

tive impacts on fish health, pollutants can also have a more indirect impact on another natural

stressor, parasites. High pollutant concentrations can lead to a lowered immune response, re-

sulting in higher parasitic infection. On the other hand, pollutants can also negatively affect

free living parasite stages, thereby reducing parasite abundance. In this study, I assess the as-

sociation between pharmaceuticals and parasite infection in a natural brown trout (Salmo trut-

ta m. fario) population. Fish were obtained from the Zivny stream (Czech Republic) upstream

(control) and downstream (polluted) of a sewage treatment plant known to release pharmaceu-

ticals. Fish length and condition parameters (condition factor, hepatosomatic index, spleen

somatic index, gonadosomatic index) did not differ between sites. Of the 79 pharmaceuticals

measured, 42 were detected in fish tissue, with highest concentrations found in liver, followed

by kidney and brain. Antibiotics and antidepressants dominated at both localities, with con-

centrations significantly higher at the downstream polluted locality (together with beta-

blockers). CNS stimulants showed similar concentrations at both sites. There was a negative

relationship between overall pharmaceutical load and fish condition factor and hepatosomatic

index at the polluted locality, while parasite abundance increased with overall pharmaceutical

load at both localities. Fish were infected by four parasite species: two monogeneans Gyro-

dactylus derjavinoides and Gyrodactylus truttae, one nematode Salmonema ephemeridarum

and one trematode Crepidostomum metoecus. Ectoparasite abundance was higher at the pol-

luted site, while endoparasite abundance was higher at the control site. The higher number of

gyrodactylids at the polluted site probably reflects an increased parasite reproduction rate un-

der stressful conditions. As macrozoobenthos density (including the intermediate hosts of the

target parasites) was higher at the polluted locality, we hypothesise that the decrease in endo-

parasites potentially resulted from as yet unidentified negative effects of pharmaceutical con-

taminants on the parasite’s free living stages.

Acknowledgements: This study was supported by the Czech Science Foundation, with project

ECIP (P505/12/G112), and by the Ministry of Education, Youth and Sports of the Czech Re-

public with projects CENAKVA (No. CZ.1.05/2.1.00/01.0024) and CENAKVA II (No. LO1205

under the NPU I program).

Abstracts

41

Future of multiplexing

N. Reslová1,2, V. Huvarová2, L. Škorpíková1, M. Kašný1, P. Králík2


2 Department of Food and Feed Safety, Veterinary Research Institute, Brno, Czechia

xMAP technology based on magnetic microspheres represents an open architecture platform,

capable of development and adaptation into many multiplex detection assays. Technology is

beneficial to end users and therefore it is utilized in various pharmaceutical, clinical and re-

search laboratories not only for nucleic acid assays but also for protein assay formats. On the

market, there are available few commercial detection panels, however, these are mainly meant

for major clinically important pathogens, infectious diseases or human health. Our research has

been focused to fill in the market gaps and develop an in-house assay suitable for detection of

genomic DNA of food-borne pathogens (e.g. Trichinella spiralis, Toxoplasma gondii, Giardia

intestinalis, Yersinia enterocolitica, Campylobacter jejuni, Salmonella enterica) in various

food-related matrices and design the diagnostic system based on the recombinant proteins char-

acteristic for pathogen infections (e.g. Trichinella spp.). Up to this date, we have optimized the

working protocol for highly specific multiplex oligonucleotide ligation PCR (MOL-PCR) to-

gether with MagPix analysis intended for multiplex detection panel for food-borne pathogens,

which is currently still being expanded. The protein assay is under optimization.

Acknowledgements: the work was supported by the Ministry of Education, Youth and Sports

of the Czech Republic (LO1218), by the Ministry of Agriculture of the Czech Republic

(RO0516), Ministry of the Interior of the Czech Republic (VI20152020044), and by Masaryk

University institutional support (MUNI/A/0816/2017).

Abstracts

42

Laser microdissection and mass spectrometry:

protein analysis of strictly selected tissue from Eudiplozoon nipponicum

P. Roudnický1, T. Loja2, D. Potěšil3, Z. Zdráhal3, M. Gelnar1, M. Kašný1


2 CEITEC, Core Facility Genomics, Masaryk University, Brno, Czechia 3 CEITEC, Core Facility Proteomics, Masaryk University, Brno, Czechia

Laser microdissection is a combined microscopic-separation technique enabling to extract the

samples of interest from histological slides - single cells, organs and tissues. The isolated

samples can be further analysed and various parameters determined. The adoption of mass

spectrometry analysis could give us the detail information about the sample composition and

relative quantity of particular components (e.g. protein molecules) can be calculated too.

Moreover, this technology enables the fast determination of distribution of selected molecule

within the studied biological object. Thus, we can avoid the time-consuming preparation of

specific antibodies in laboratory animals, which are widely used for protein tissue localiza-

tion. In our current research, we used Eudiplozoon nipponicum, ectoparasitic blood-feeder

from gills of Cyprinus carpio with aim to reveal the protein composition of its tegument re-

gion, gut and inner parenchyma. So far, we have identified 405 unique proteins in inner pa-

renchyma/gut and 131 unique proteins in tegument. Among them, some already described

functional molecules have been found.

Acknowledgements: The research was financially supported by Czech Science Foundation

(GBP505/12/G112) and Masaryk University (MUNI/A/0816/2017).

Abstracts

43

Metabolism of anthelmintics and drug resistance in helminths

P. Matoušková, I. Vokřál, L. Raisová Stuchlíková, J. Lamka, B. Szotáková, L. Skálová

Faculty of Pharmacy, Charles University, Hradec Králové, Czechia

In all organisms, drugs as well as other xenobiotics undergo metabolic transformation via

action of drug metabolizing enzymes (DME) i.e. drug transporters and biotransformation en-

zymes. Generally, DME transform drug into less biologically active and more easily elimina-

ble metabolites. By this way, DME serve as an efficient defense against the potentially nega-

tive action of xenobiotics and the activities of DME determine both desired and undesired

effects of drugs. While human and mammalian DME have been intensively studied, DME of

helminths parasites have been almost neglected for many years. Nowadays, the research of

helminths´ DME has been more intense as the relationship between DME and drug resistance

has been considered. In stable resistant strains, the resistance might be caused by changes in

genes for DME or in corresponding control (regulatory) segments. In sensitive strains, the

helminth DME may represent a tool contributing to the development of resistance as the ac-

tive efflux of a drug together with its deactivation via biotransformation diminishes drug effi-

cacy and may allow some worms to survive the anthelmintic therapy. In our labs, we studied

the metabolism of several anthelmintics (e.g. albendazole, fenbendazole, flubendazole,

triclabendazole, ivermectin, monepantel) in adults of Haemonchus contortus, Dicrocoelium

dendriticum, Fascioloides magna, Fasciola hepatica, Hymenolepis diminuta, Moniezia ex-

pansa. In most cases, helminths were able to metabolize the anthelmintics with H. contortus

being the most metabolically active. Several helminth–specific metabolites of anthelmintics

were identified. When metabolism of anthelmintics was compared in H. contortus strains sen-

sitive and resistant to anthelmintics, significant differences were found. The adults of the re-

sistant strains were able to form many more metabolites of all the drugs than the adults of the

sensitive strain. Some metabolites were even found only in adults of the resistant strains.

These findings suggest that increased drug metabolism is one of the mechanisms of drug re-

sistance in H. contortus.

Acknowledgements: This project is supported by Czech Science Foundation, grants No. 17-

11954Y and 18-07724S.

Abstracts

44

Identification of immunogenic proteins in excretory-secretory products

of Toxocara canis larvae

K. Skulinová1, J. Novák1, L. Kolářová1,2, M. Kašný3,4

1 Institute of Immunology and Microbiology, First Faculty of Medicine,

Charles University, Prague, Czechia 2 National Reference Laboratory for Tissue Helminthoses, General University Hospital in

Prague, Prague, Czechia 3 Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czechia


Larval toxocarosis (LT) is a cosmopolitan zoonosis caused by third-stage larvae of nematodes

from genus Toxocara, intestinal parasites of dogs, cats and foxes. Humans are infected by

ingestion of eggs persisting in soil or on unwashed fruits and vegetables, but also by eating

undercooked meat of other paratenic hosts containing encapsulated larvae. Disease results in a

broad spectrum of clinical manifestation influenced by various factors. Routine serodiagnos-

tics of LT is based on the detection of specific antibodies by ELISA using T. canis excretory-

secretory product (TES) as a pooled antigen, obtained by in vitro cultivation. The objective of

our research is to characterize the immunodominant TES protein components and select the

particular specific reliable immunogenic protein candidates for further preparation of their

recombinant forms, which could be used in routine serodiagnostics of human larval toxocaro-

sis. Thus, would be possible to avoid the laborious process and necessity to use the unlimited

source of T. canis females. The medium with TES antigens was periodically collected (1-14

days), followed by purification and concentration of protein fraction. SDS-PAGE electropho-

retic analysis was performed to reveal TES protein composition, followed by Western blot.

PVDF membrane containing separated TES was incubated with positive and negative mice

sera and anti-Toxocara IgG antibodies-Px. Protein bands corresponding to the area of reaction

were cut off from the gel and analyzed by mass spectrometry. The evaluation of results is in

process and until now we have identified several auspicious candidates for further investiga-

tion.

Acknowledgements: The study was supported by the Charles University, projects GA UK

(902217) and SVV (260 369).

Abstracts

45

Evaluating the role of trematode cercariae in the energetics of a subarctic ecosystem

M. Soldánová1, A. Born-Torrijos1, J. Schwelm2, G. S. van Beest1,3, T. Vyhlídalová4, E. H.

Henriksen5, R. Knudsen5, R. Kristoffersen5, P.-A. Amundsen5


České Budějovice, Czechia 2 Aquatic Ecology and Centre for Water and Environmental Research,

University of Duisburg-Essen, Essen, Germany 3 Cavanilles Institute for Biodiversity and Evolutionary Biology, Science Park,

University of Valencia, Spain 4 Faculty of Science, University of South Bohemia in České Budějovice, Czechia

5 Department of Arctic and Marine Biology, Faculty of Biosciences, Fisheries and Economics,

UiT The Arctic University of Norway, Tromsø, Norway

Free-living stages of trematodes (Digenea), the cercariae, are essential components of aquatic

food webs. A high number of cercariae can be daily released from their molluscan hosts and

may thus generate a substantial biomass within an ecosystem, potentially being important in

the energy flow through the food web as prey items for free-living species. However, infor-

mation on the role of trematode cercariae in aquatic food webs is very scarce and completely

missing for subarctic freshwater systems. In this study, we evaluated the ecological im-

portance of cercariae and the strengths of their trophic interactions by obtaining data and es-

timates of (i) cercarial emission, output rates and biomass, (ii) survival rates of cercariae, and

(iii) predation rates towards cercariae from direct and indirect (i.e. concomitant) predation by

different aquatic animals. Although these aspects have been studied separately, they have

never been combined and thus remain to be tested jointly. Sampling of cercariae from natural-

ly infected snails Radix balthica and tentative predators upon these cercariae was conducted

in two seasons in the subarctic Lake Takvatn, northern Norway. In a series of field and la-

boratory experiments, several trematode species and plausible vertebrate and invertebrate

predators were used. Our preliminary results on trematode production and parasite-predator

interactions reveal a considerably high cercarial biomass potentially present within the lake

ecosystem. However, the cercarial biomass can be significantly reduced by extensive con-

sumption by predators, as cercariae may serve directly as a food resource or are indirectly

consumed along with their snail host (i.e. concomitant predation). Despite the general consid-

eration that parasites are small and thus negligible in terms of biomass contribution, our study

suggests that cercariae of trematodes may play a significant role with a potential to transfer

substantial biomass within food webs and ecosystems.


17-20936Y), Institute of Parasitology, Biology Centre of the Czech Academy of Sciences

(RVO: 60077344) and UiT The Arctic University of Norway and the Norwegian Research

Council (Project No. 213610)

Abstracts

46

Characterization of egg and miracidium ultrastructures of two schistosome species

with regard to their different life strategies

K. Svobodová, P. Horák


The avian flukes Trichobilharzia regenti and T. szidati belong to the family Schistosomatidae.

They are dixenous trematodes bound to the aquatic environment, with similar definitive hosts.

The free-living larvae called cercariae penetrate the skin and transform to schistosomula not

only in the avian hosts, but also in the accidental mammalian hosts including humans; they

may cause inflammatory skin reaction called cercarial dermatitis (or swimmer’s itch). In ver-

tebrates, the life strategy of these two species differs. Trichobilharzia regenti is a unique

among schistosomes due to its high affinity to the nervous tissue. In ducks, the transformed

schistosomula migrate via the peripheral nerves and the central nervous system to the nasal

mucosa, where they mature to adults, copulate and lay eggs. Usually, the eggs of avian schis-

tosomes, after maturation in and exit from the host, contain fully developed miracidia. Sur-

prisingly, the miracidia of T. regenti hatch directly in the nasal mucosa and leave the host to

the aquatic environment. Trichobilharzia szidati is a visceral avian schistosome. It means that

schistosomula migrate to the veins and tissues of visceral organs where they reach sexual ma-

turity, copulate and lay eggs. The eggs leave the avian host with the feces, and miracidia hatch

soon from the eggs in water reservoirs. Until now, there is no information on morphological

and physiological differences between these two species regarding miracidia hatching. There-

fore, the primary goal of our research is to characterize the ultrastructure of the eggs and the

miracidia of these two avian flukes, and search for ultrastructural features that could clarify

the different mode of hatching. Transmission microscopy will be used for this purpose.

Abstracts

47

Differentiation of Eight Trichinella Species Using a High Resolution Melting Assay.

L. Škorpíková1,2, N. Reslová1,2, E. Pozio3, M. Kašný1

1 Department of Botany and Zoology, Masaryk University, Brno, Czechia

2 Department of Food and Feed Safety, Veterinary Research Institute, Brno, Czechia 3 Department of Infectious Diseases, Istituto Superiore di Sanità, Rome, Italy

Cosmopolitan nematodes of the genus Trichinella are foodborne parasites with the high im-

pact on animal trade and food safety. A broad range of carnivore and omnivore animals have

been reported to be hosts of these parasites, including the economically important domestic

pig. The representatives of the genus Trichinella are also causative agents of human trichinel-

losis, a serious human disease, which has been documented at least in 55 countries around the

world. Based on zoogeographical, epidemiological and molecular characters, there are 12 taxa

recognised in the genus Trichinella. According to ability of muscle larvae to encapsulate in

host muscle tissues or not, two main clades (encapsulated and non-encapsulated) are recog-

nized. Despite this difference in capsule formation, there are no clear morphological features

useful for species differentiation. In this study, we adopted a high resolution melting analysis,

a single-tube method, which can be carried out as an additional step following a real-time

quantitative PCR. This method enables the distinguishing of genetic variation (down to single

nucleotide polymorphisms) in amplified DNA fragments without necessity of subsequent se-

quencing. Our approach differentiating capacity was evaluated using eight Trichinella species

(T1, T2, T3, T4, T5, T7, T10 and T11), 37 samples of genomic DNA isolated from single

muscle larvae were analyzed. The assay was based on polymorphisms in 240 bp long frag-

ment of the cytochrome c oxidase subunit I gene. This mitochondrial gene shows high se-

quence divergence even among closely allied species. For the evaluation of melting process

have been used difference plot, which represents the most transparent form of expression of

species-specific matrix curves. We show that high resolution melting analysis is rapid and

reliable method that can be used to identify single larvae of eight Trichinella species. In the

future we envisage that Trichinella species-specific matrix curves could be automatically

evaluated using the computer application.

Acknowledgements: The study was financially supported by Masaryk University institutional

support (MUNI/A/0816/2017) and grants from the Ministry of Agriculture (RO0517) and the

Ministry of Education, Youth and Sport (LH12096, LO1218 and LD15056).

Abstracts

48

Production of nitric oxide in mice infected by Trichobilharzia regenti (Schistosomatidae)

B. Šmídová, T. Macháček, P. Horák


The neuropathogenic avian schistosome Trichobilharzia regenti is not able to complete its life

cycle in mammals, such as mice. Production of nitric oxide (NO) might be a reason for this

host incompatibility. NO produced by inducible NO synthase (iNOS) affects cell structures in

both parasite and host cells; among others, it binds to tyrosine resulting in 3-nitrotyrosine (3-

NT) formation. Our aim was to monitor the production of NO in infected mice 3, 7, 14 and 21

days post infection (dpi); uninfected mice were used as negative control. First, we measured

nitrite/nitrate concentration in serum samples of infected mice by Griess reaction. Second, we

applied immunohistochemistry to detect iNOS and 3-NT in the murine spinal cords. Prelimi-

nary data show rather no increase of nitrites/nitrates in the serum. However, a certain trend in

iNOS and 3-NT expression has been noticed in the injured spinal cords, which has to be fur-

ther confirmed.

Acknowledgement: This study was supported by Charles University Grant Agency (project no.

729516) and by Czech Science Foundation (18-11140S).

Abstracts

49

Molecular determination of Trichobilharzia spp. DNA in the body fluids of their hosts

V. Šteiger1, P. Horák1, M. Kašný1,2



The life cycle of the genus Trichobilharzia covers the intermediate host - pulmonary water

snails (mostly the family Lymnaeidae) and the definitive host – birds (mostly the order Anser-

iformes). The first larva, miracidium, after release from the egg, actively moves in water reser-

voirs, penetrates the snail and develops to mother sporocyst. Then the daughter sporocysts are

formed and migrate to the hepatopancreas of the snail where the huge amount of cercariae is

asexually produced. Cercariae leave the intermediate host and penetrate the skin of definitive

host. Within the host body they transform to schistosomula and migrate to the final location

where they mature and lay eggs. Cercariae can also penetrate the skin of some mammals (in-

cluding humans) and cause an inflammatory reaction called cercarial dermatitis (swimmer’s

itch). Until now, for humans, there is no effective method enabling to differ cercarial dermatitis

from other hyper-immune skin reactions and for birds the gentle diagnostic method of trichobil-

harziasis is missing. The aim of our study was to design a new molecular method focused on the

detection of cell-free DNA in the body fluids of the host. In our experiments we used two ex-

perimental parasites belonging to the genus Trichobilharzia, the neurotropic T. regenti and the

visceral T. szidati. To reveal the presence of parasite we chose 396 bp tandem repeated DNA

sequence (highly represented in the genome of the fluke) as a molecular marker and samples of

the snail hemolymph and the duck/mice blood and nervous tissue. Due to our effort to optimize

the method, we tried to detect T. regenti DNA in the hemolymph of infected snails Radix lago-

tis which were dissected 36 and 66 days after infection. Using the conventional PCR, we were

successful to amplify the parasite DNA in all samples from the infected snails, whereas and no

amplicon was observed in the group of uninfected snails. The same result was recorded also for

DNA samples originating from the parasite-host system T. szidati and Lymnaea stagnalis. Next,

we tried to adopt this approach also for qPCR-based detection of bird schistosome DNA in the

blood samples from definitive and accidental hosts. We recorded only a partial success, because

the parasite DNA was detected only in some samples.

The work was supported by the Czech Science Foundation (No. 13-29577S and No. 18-

11140S) and the Charles University in Prague (grant No. GAUK 243259305).

Abstracts

50

Glycomic analysis of developmental stages of the neurotropic schistosome

Trichobilharzia regenti

L. Turjanicová1, D. L. Nguyen2, A. van Diepen2, C. H. Hokke2, P. Horák1, L. Mikeš1

1Department of Parasitology, Faculty of Science, Charles University, Prague, Czechia 2Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands

All organisms including parasites produce carbohydrate structures, generally called glycans.

Parasite glycan structures are often immunogenic, affect the development of immune respons-

es in the host, and serve as mediators in the interactions between parasites and their hosts. The

main aim of our research was to study glycans of two life stages (cercariae and schistosomula)

of the avian neurotropic schistosome Trichobilharzia regenti. Comparison with glycan spectra

of human schistosomes, especially of Schistosoma mansoni, has also been done. We analyzed

the pattern of surface glycan structures in T. regenti by binding of specific anti-glycan mono-

clonal antibodies developed on the basis of various glycan moieties of S. mansoni. The mAbs

recognized DF motif (Fucα1-2Fucα1-), LDN-DF motif (GalNAcβ1-4(Fucα1-2Fucα1-

3)GlcNAcβ1-), and F-LDN motif (Fucα1-3GalNAcβ1-4GlcNAcβ1-) on the surface of cercar-

iae. On the other hand, the surface of 7 days old schistosomula was recognized by anti-LDN-F

(GalNAcβ1-4(Fucα1-3)GlcNAcβ1-) and Anti-LeX (Galβ1-4(Fucα1-3)GlcNAcβ1-) mAbs.

Furthermore, we released N- (Asn) and O-linked (Ser/Thr/ceramide) glycans from glycopro-

teins and glycolipids from parasite homogenates and analyzed them by a mass spectrometry-

based glycomics approach. The N-glycan fractions from cercariae contained predominantly

oligomannose glycans. Other dominant motifs usually found on the branches of N-glycans

comprised LDN (GalNAcβ1-4GlcNAcβ1-) and LN (Galβ1-4GlcNAcβ1-). The oligomannose

structures were preserved in 7-day old schistosomula, while expression of LND glycan motifs

declined, and expression of LeX-motifs increased. Our study represents new results, which

may set the foundation for understanding how glycans influence different aspects of interac-

tions between bird schistosomes and their hosts.

Acknowledgements: The study was supported by Czech Science Foundation (Grant Nos. 13-

29577S and 18-11140S) and by the project “Centre for Research of Pathogenicity and Viru-

lence of Parasites” (no. CZ.02.1.01/0.0/0.0/16_019/0000759) funded by European Regional

Development Fund and Ministry of Education, Youth and Sports of the Czech Republic.

Charles University institutional support (PROGRES Q43, UNCE 204017, and SVV 244-

260432/2017) applied to LT, PH, and LM.

Abstracts

51

New insights into the scolex morphology of monozoic cestodes (Caryophyllidea)

from the Nearctic Region

D. Uhrovič1, M. Oros1, T. Scholz2

1 Institute of Parasitology, Slovak Academy of Sciences, Košice, Slovakia



A comparative study of the scoleces of monozoic tapeworms (Cestoda: Caryophyllidea), par-

asites of catostomid and cyprinid fishes (Teleostei: Cypriniformes) in the Nearctic Region,

was carried out. The scolex of caryophyllidean tapeworms is highly motile and variable in

shape with no single type being characteristic of this group. Using light and scanning electron

microscopy, the scoleces of representatives of 19 genera of North American caryophyllideans

are characterised. Whenever possible, material newly collected by the present authors and

processed using the same method (hot fixative) to ensure comparability of morphological and

biometrical data was preferentially used. In addition, data based on the examination of type

specimens of numerous taxa were also considered. The following basic scolex types can be

recognised among Nearctic taxa: papillate, loculopapillate and loculotruncate (Promonoboth-

rium), loculopapillate (Dieffluvium), bothrioloculodiscate (Archigetes, Janiszewskella, Penar-

chigetes and Pseudoglaridacris), fixomegabothriate (Capingens), bulbate and bulboacuminate

(Atractolytocestus), cuneiloculate (Spartoides, Rowardleus and Hypocaryophyllaeus), bulb-

oloculate, bothrioloculodiscate (Biacetabulum), tholate (Hunterella), cuneifimbriate (Khaw-

ia), cuneiform (Calentinella, Caryophyllaeides and Edlintonia), hastate (Pseudolytocestus),

loculotholate (Bialovarium and Pliovitellaria) and cuneiformoloculate (Glaridacris and Iso-

glaridacris). Considerable variation in scolex morphology exists among species of some gen-

era such as Promonobothrium Mackiewicz, 1968, whereas species of others such as Biac-

etabulum Hunter, 1927 have the scolex of rather uniform shape. Unique is the scolex of the

genus Capingens Hunter, 1927, which is massive, robust, with large and deep bothria. The

structure of the so called acetabulum in species of Biacetabulum, supposedly homologous

with acetabulate attachment organs of more recently evolved groups of tapeworms (bothridia

and suckers), has been critically examined using histological sections. This study has revealed

that this attachment structure in fact correspond to loculi, not to any acetabulate structure,

which has to be delimited by a distinct layer of muscle fibres. The present study has con-

firmed high variability of scolex types in the Nearctic species of caryophyllidean cestodes

compared with those from other zoogeographical regions.

The study was financially supported by the Grant Agency VEGA (No. 2/0159/16) and Czech

Science Foundation (project No. P505/12/G112).

Abstracts

52

Interactions between Trichobilharzia regenti and duck definitive host in the terminal

phase of infection: histological evaluation

L. Vlčková, P. Horák


Trichobilharzia regenti is a dixenic avian fluke belonging to Schistosomatidae (Trematoda).

The parasite ends its unique journey through the central nervous system in the nasal mucosa

of waterfowl. The adults are located either in the blood system or mostly freely in the connec-

tive tissue, where they lay eggs in clusters. Unlike human schistosomes, the fully developed

miracidium hatches right in the tissue, and leaves the host. We can observe massive infiltra-

tion of cells and accumulation of amorphous mass around the eggs, destruction and fibrotic

reparation of the tissue accompanied by hemorrhage. Whereas no immune reaction can be

detected around the adults, it seems that an inflammation is present in the egg vicinity. Our

results determined the eggs as the main causative agent of histopathological changes in the

tissue just like in human schistosomiasis. Histological methods showed granulocytes, mono-

nuclear cells, mastocytes and collagen fibres around the eggs. Immunohistochemical (IHC)

probes showed cells positive for 3-nitrotyrosine (3-NT), a marker of cell damage and inflam-

mation formed in the presence of nitric oxide (NO). The accumulation of 3-NT-positive cells

was observed around some eggs 16 day post infection (dpi), and some cells were also present

around the adults. Single 3-NT-positive cells were diffused in the tissue. We also observed

(16 dpi) some oxidized structures that were sporadically distributed in the tissue, in the amor-

phous mass around the eggs in some cases, and on the cells located intravascularly. As for the

markers on immune cells, we presuppose the presence of BAFF+ cells because we noticed

some signals using specific antibody. Nevertheless, there were some other cells, mostly gran-

ulocytes, and possibly CD4+ and KUL01+ cells that showed a stronger signal. Amorphous

mass around the egg clusters was KUL01+. It seems that the eggs and even the adults are sur-

rounded by an inflammation, and later by fibrosis.

Abstracts

53

Efflux transporters in Haemonchus contortus – role in drug resistance development

I. Vokřál1, P. Matoušková2, H. Danková1, B. Szotáková2, L. Skálová2

1 Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové,

Charles University, Prague, Czechia 2 Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové,

Charles University, Prague, Czechia

Efflux transporters protect living organisms against toxic xenobiotic compounds including

drugs. Five district families of efflux transporters exist from which ATP-binding cassette

transporters family is the most important. These transporters were already identified in

roundworms, flukes and tapeworms. In roundworms, efflux transporters are suspected to con-

tribute to the worldwide emerging drug-resistance development, predominantly to macrocy-

clic lactones. From this perspective, the most studied efflux transporter is P-glycoprotein and

its homologs. In Haemonchus contortus, parasitic roundworm of ruminants, resistance to all

common anthelmintic drugs (e.g. benzimidazole drugs, macrocyclic lactones or amino-

acetonitrile derivatives) was already described. H. contortus has several P-glycoprotein homo-

logs and it is not fully known which are connected to the drug resistance development. Aim of

our study was to compare expression of selected P-glycoprotein homologs (Pgp-1, 2, 9.1, 9.2,

11, 16) in adult stage of two H. contortus resistant strains, WR (multi-resistant) and IRE

(ivermectin-resistant). Emphasis was put also on sex differences. Our preliminary results

show differences in expression of efflux transporters not only between the strains, but also

between males and females of both strains.

Acknowledgements: This project was supported by Czech Science Foundation (Grant No.

17-11954Y) and by Charles University (PRIMUS/17/SCI/4 and SVV 260 416)

Abstracts

54

Proteomic analysis of helminth tissues using microdissection

– from cryosections to raw data

O. Vondráček1, P. Talacko2, R. Leontovyč1, J. Bulantová1, P. Horák1


2 Proteomics Core Facility, Faculty of Science, Charles University, BIOCEV, Vestec, Czechia

To understand the various parasite-host interactions, such as invasion, migration or nutrition,

the knowledge of protein composition of selected parasite tissues/organs can be of help. Our

aim is to reveal the functional characterization of the protein composition of selected tissues of

Trichobilharzia larvae. For example, we are going to compare intestinal proteins of two species,

namely T. regenti and T. szidati, which have different feeding preferences and migratory routes

in the vertebrate host. The basis for a good proteomic analysis is to obtain sufficient amount of

material without contamination from the surrounding tissues. Unfortunately, the small size and

the body architecture of these parasites represent a general problem. Moreover, in proteomics,

unlike in other molecular analyses, the proteins are not amplified prior to their analysis. Nowa-

days, these obstacles can be overcome using laser microdissection techniques.

Our approach includes microdissection of cryosections (using MMI CapLift technology) fol-

lowed by one-step reduction and alkylation with subsequent protein cleavage prior sample

analysis by LC-MS/MS (Orbitrap Fusion system). After evaluation of raw data, the selected

proteins should be backtracked using immunohistochemistry or in situ hybridization.

In this work we dealt with the optimization of a procedure that precedes the proteomic analy-

sis. Using T. szidati cercariae we tested (a) the impact of fixatives, (b) the effect of sample

processing after microdissection, (c) the optimal amount/volume of the analyzed material, and

(d) the time-dependent material degradation before LC-MS/MS analysis.

The use of 7.5 mils µm3 of unfixed (frozen in the optimal cutting temperature (OCT) com-

pound) microdissected material processed via the In-StageTips method provided the best re-

sults. In addition, the exposition of the unfixed microdissected material to room temperature

up to 24 hours (simulating a long microdissection process) had no significant effect on the

proteomic analysis.

Acknowledgement: The study was supported by PROGRES Q43, UNCE 2018/012, SVV

260432/2018, Czech Science Foundation (18-11140S) and by European Regional Develop-

ment Fund (CZ.1.05/1.1.00/02.0109; CZ.02.1.01/0.0/0.0/16_019/0000759).

Abstracts

55

Monogeneans in the pores

J. Vorel1, M. Jankůjová2, J. Oppelt2, F. Pardy3, P. Roudnický1, J. Ilgová1,

L. Mikeš4, M. Gelnar1, M. Kašný1


2 National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czechia 3 Central European Institute of Technology, Faculty of Science, Masaryk University, Brno, Czechia


Ectoparasitic flatworms from the group Monogenea represent serious fish pathogens. Their

presence in stocks can lead to significant losses in fish host populations. Despite this fact, the

running research has been focused mainly on morphological and phylogenetical characteris-

tics of these worms and the information related to the biochemical and molecular nature of the

physiological processes is rather sporadic. Up to now, we performed whole-genomic sequenc-

ing of selected monogenean representative Eudiplozoon nipponicum by three sequencing

techniques. 454/Roche (Junior sequencing platform), Illumina (MiSeq and HiSeq sequencing

platforms) and recently by Oxford Nanopore (MinION sequencing platform). Using 454/Ro-

che Junior and both Illumina sequencing platforms, 164,773,962 short genomic reads were

originally generated. After quality processing of raw reads (trimming, removing contami-

nants, errors correction, etc.), 130,741,241 reads were used for the draft of genome assembly.

Finally, 524,914 contigs were generated by MaSuRCA assembler in total length

1,109,208,298 base pairs (1.1 Gb). Unfortunately, the final assembly was fragmented and

significant number of contigs was identified as a bacterial contamination. In order to improve

E. nipponicum draft genome, especially to fill the missing genome gaps, we generated long

reads using modern third generation sequencing method nanopore sequencing (MinION). The

MinION platform (Oxford Nanopore Technologies, UK) is a small and portable, real time

working, low-cost USB device, producing very long-reads (theoretically up to 250 Kb). The

results of our analysis with E. nipponicum DNA showed, that 500 active channels/pores (from

max. 512) produced 1,091,445 reads with length up to 198,101 base pairs (average 2,588 base

pairs). In total, more than 2,8 Gb were generated in two days run, it significantly helped us to

improve genome information by merging a relevant percentage of contigs (9.7 %), from

524,914 to 474,134.

Acknowledgements: The research was financially supported by Czech Science Foundation

(GBP505/12/G112) and Masaryk University (MUNI/A/0816/2017).

Abstracts

56

Diversity of diplozoid species (Monogenea, Diplozoidae) parasitizing the gills

of endemic cyprinids from Mediterranean area

K. Vyčítalová, B. Koubková, A. Šimková

Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czechia

The Mediterranean area is known for its exceptional endemic fauna including cyprinid spe-

cies. Endemic cyprinids represent one of the most diverse freshwater fish groups in this area,

but knowledge of their parasitofauna is very limited. In this study, a total of 125 endemic cy-

prinid species were investigated in 2014 – 2017 for the presence of diplozoid monogeneans

parasitizing the gills of cyprinids in the selected areas of Mediterranean. Identification of dip-

lozoid species was performed using morphological analysis of the attachment apparatus and

molecular analysis using ITS2 region. Phylogenetic analysis of the partial ITS2 together with

morphological data revealed five species of Paradiplozoon. Paradiplozoon homoion, com-

mon parasite of several species of cyprinids in Central Europe, was the most abundant species

in endemic cyprinids of Mediterranean. This study provides new records of host species for P.

homoion – 12 from localities in Balkan Peninsula, e.g. Chondrostoma knerii, Scardinius

plotizza or Telestes beoticus and 5 from the localities in Spain, e.g. Parachondrostoma miegii

or Squalius laietanus. Paradiplozoon megan is known as a parasite of Squalius and Leuciscus

in Central Europe. We determined P. megan on Squalius zrmanjae and S. squalus from Croa-

tia and S. lucumonis, S. squalus, Protochondrostoma genei and Rutilus rubilio from Italy. In

addition, we found three species of Paradiplozoon representing potentially new species for

science. Paradiplozoon sp. 1 parasitized Scardinius acarnanicus and Tropidophoxinellus hel-

lenicus from Greece. This species is phylogenetically closely related to P. megan. Paradiplo-

zoon sp. 2 was recorded on 9 species of different genera of cyprinids in Iberian Peninsula, i.e.

Iberochondrostoma, Parachondrostoma, Pseudochondrostoma, Luciobarbus and Squalius.

This species is phylogenetically related with P. pavlovskii. Paradiplozoon sp. 3 was reported

only on Moroccan Luciobarbus lepineyi. This species is phylogenetically closely related to P.

bingolensis from Garra rufa (Turkey).

Acknowledgements: This study was financially supported by Grant Agency of the Czech Re-

public, project No. 15-1938S and since 2018 is partially supported by the ECIP project no.

P505/12/G112.

Abstracts

57

Production and periodicity in the emergence of cercariae of Diplostomum spp. (Digenea)

in the lakes of temperate and subarctic regions

T. Vyhlídalová1, M. Soldánová2

1 Faculty of Science, University of South Bohemia in České Budějovice, Czechia



Free-living stages of trematodes, the cercariae, are recognised as an important part of energy

flow in food webs within aquatic ecosystems because cercarial production from infected first

intermediate molluscan host is substantial in most host-trematode systems. Cercarial emission

patterns generally depend on various abiotic and biotic factors of the environment, including

temperature, light or behaviour of hosts to maximise transmission success. Trematodes of the

genus Diplostomum spp. (Diplostomidae) are widely distributed pathogens of fish causing eye

cataracts that may lead to host mortality. Although the production and chronobiology in cer-

carial emergence of some species of Diplostomum have been investigated, most information

comes from laboratory conditions using large lymnaed snail host such as Lymnaea stagnalis,

whereas other host-trematode systems are poorly studied. In this study we investigated the

effects of photo- and thermoperiod on the emission of Diplostomum cercariae emerging from

Radix spp. in two geographical areas with distinctly different light and temperature climate

conditions. Cercariae were obtained from snails collected in Lake Takvatn in northern Nor-

way and lakes Most and Medard in northern Bohemia, Czechia. Both field and laboratory

experiments were conducted in two periods of the year (summer and autumn) following the

natural light/dark and temperature conditions directly in the lake and in the environmental

chamber, simulating natural sunlight regimes. Additionally, experiments in the temperature

5°C higher that in the field were performed to assess cercarial sensitivity to temperature

changes. Cercarial output was measured over 24-hour period for 3 consecutive days. Prelimi-

nary data indicate contrasting patterns in cercarial patterns in emergence depending on the

geographical latitude with different light and temperature conditions. The emission strategies

of cercariae of Diplostomum in subarctic areas indicate a stronger adaptive mechanism that

may compensate reduced possibilities in narrow transmission window under extreme subarc-

tic climate. Our data underline the importance to study the interplay between environmental

factors and parasite life strategies.


17-20936Y), the Institute of Parasitology, Biology Centre of the Czech Academy of Sciences

(RVO: 60077344) and the Department of Parasitology, Faculty of Science, University of

South Bohemia in České Budějovice.

Participant contact information

58

PARTICIPANT CONTACT INFORMATION

Alves Philippe, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31, 370

05 České Budějovice, Czechia, [email protected]

Barčák Daniel, Institute of Parasitology SAS, Hlinkova 3, 040 01 Košice, Slovakia, bar-

[email protected]

Benovics Michal, Department of Botany and Zoology, Faculty of Science, Masaryk Universi-

ty, Kotlářská 2, 611 37 Brno, Czechia, [email protected]

Bernardová Nicol, Institute of Immunology and Microbiology, First Faculty of Medicine,

Charles University and General University Hospital in Prague, Studničkova 7, 128 00 Praha

2, Czechia, [email protected]

Bulantová Jana, Department of Parasitology, Faculty of Science, Charles University,

Viničná 7, 128 44 Praha 2, Czechia, [email protected]

Čenková Miroslava, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31,

370 05 České Budějovice, Czechia, [email protected]

De Oliveira Camila, Federal Rural University of Rio de Janeiro, BR 465, Seropédica, Rio de

Janeiro, Brazil, [email protected]

Dimunová Diana, Department of Biochemical Sciences, Faculty of Pharmacy, Charles Univer-

sity, Akademika Heyrovského 1203, 500 05 Hradec Králové, Czechia, [email protected]

Dvořák Jan, Czech University of Life Sciences and Institute of Organic Chemistry and Bio-

chemistry, CAS, v.v.i., Kamýcká 961/129, 165 00 Praha-Suchdol, [email protected]

Faltýnková Anna, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31,


Gelnar Milan, Department of Botany and Zoology, Faculty of Science, Masaryk Universi-


Havelková Markéta, Department of Parasitology, Faculty of Science, Charles University,


Heinclová Pavla, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31, 370


Horák Petr, Department of Parasitology, Faculty of Science, Charles University, Viničná 7,

128 44 Praha 2, Czechia, [email protected]

Hyršl Pavel, Institute of Experimental Biology, Faculty of Science, Masaryk University, Ko-

tlářská 2, 611 37 Brno, Czechia, [email protected]

Chanová Marta, Institute of Immunology and Microbiology, First Faculty of Medicine,

Charles University and General University Hospital in Prague, Studničkova 7, 12800 Praha 2,

[email protected]

Ježková Monika, Department of Parasitology, Faculty of Science, Charles University,


mailto:[email protected]










https://maps.google.com/?q=Kotl%C3%A1%C5%99sk%C3%A1+2&entry=gmail&source=g









59

Jirků Miloslav, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31, 370


Jirsová Dagmar, Department of Botany and Zoology, Faculty of Science, Masaryk Universi-

ty, Kotlářská 2, 611 37 Brno, Czechia, University campus Bohunice, Kamenice 5, 625 00

Brno, Czechia, [email protected]

Kašný Martin, Department of Parasitology, Faculty of Science, Charles University, Depart-

ment of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37

Brno, Czechia, [email protected]

Kmentová Nikol, Department of Botany and Zoology, Faculty of Science, Masaryk Universi-


Kolářová Libuše, Institute of Immunology and Microbiology, First Faculty of Medicine,

Charles University, Prague, Studničkova 7, 128 00 Praha 2, [email protected]

Konečný Lukáš, Department of Parasitology, Faculty of Science, Charles University,


Krasnovyd Vadym, Department of Botany and Zoology, Faculty of Science, Masaryk Uni-

versity, Kotlářská 2, 611 37 Brno, Czechia, [email protected]

Kuchta Roman, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31, 370


Leontovyč Adrian, Institute of Organic Chemistry and Biochemistry, CAS, v.v.i, Flemingo-

vo náměstí 542/2, 160 00 Praha 6, Czechia, [email protected]

Leontovyč Roman, Department of Parasitology, Faculty of Science, Charles University,


Li Liang, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31, 370 05

České Budějovice, Czechia, [email protected]

Macháček Tomáš, Department of Parasitology, Faculty of Science, Charles University,


Majer Martin, Department of Parasitology, Faculty of Science, Charles University, Viničná

7, 128 44 Praha 2, Czechia, [email protected]

Matoušková Martina, Institute of Parasitology SAS, Hlinkova 3, 040 01 Košice, Slovakia,

[email protected]

Matoušková Petra, Faculty of Pharmacy, Charles University, Akademika Heyrovského

1203, 50005 Hradec Králové, Czechia, [email protected]

Matušková Denisa, Department of Zoology and Fisheries, Czech University of Life Scienc-

es, Kamýcká 961/129, 165 00, Praha-Suchdol, Czechia, [email protected]

Mazanec Hynek, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31, 370


Mikeš Libor, Department of Parasitology, Faculty of Science, Charles University, Viničná 7,

128 44 Praha 2, Czechia, [email protected]




















60

Morand Serge, CNRS ISEM Université Montpellier - CIRAD ASTRE, Fran-

ce, [email protected]

Oros Mikuláš, Institute of Parasitology SAS, Hlinkova 3, 040 01 Košice, Slovakia, [email protected]

Pankrác Jan, Center for Advanced Preclinical Imaging, First Faculty of Medicine, Charles

University, Salmovská 3, 120 00, Praha 2, Czechia, [email protected]

Peterková Kristýna, Department of Parasitology, Faculty of Science, Charles University,


Pokrupová Zuzana, Department of Parasitology, Faculty of Science, Charles University,


Pravdová Markéta, Department of Botany and Zoology, Faculty of Science, Masaryk Uni-


Reslová Nikol, Department of Botany and Zoology, Faculty of Science, Masaryk University,

University campus Bohunice, Kamenice 5, 625 00 Brno, Czechia, Department of Food and

Feed Safety, Veterinary Research Institute, v.v.i., Hudcova 296/70, 621 00 Brno, Czechia,

[email protected]

Roudnický Pavel, Department of Botany and Zoology, Faculty of Science, Masaryk University,

University campus Bohunice, Kamenice 5, 625 00 Brno, Czechia, [email protected]

Scholz Tomáš, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31, 370 05

České Budějovice, Czechia, [email protected]

Schreiber Manfred, Department of Parasitology, Faculty of Science, Charles University,


Skálová Lenka, Department of Biochemical Sciences, Faculty of Pharmacy, Charles Universi-

ty, Akademika Heyrovského 1203, 500 05 Hradec Králové, Czechia, [email protected]

Skulinová Kateřina, Institute of Immunology and Microbiology, First Faculty of Medicine,

Charles University, Studničkova 7, 12800, Praha, Czechia, [email protected]

Soldánová Miroslava, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31,


Svobodová Kateřina, Department of Parasitology, Faculty of Science, Charles University,


Škorpíková Lucie, Department of Botany and Zoology, Faculty of Science, Masaryk Univer-

sity, University campus Bohunice, Kamenice 5, 625 00 Brno, Czechia, skorpiko-

[email protected]

Šmídová Barbora, Department of Parasitology, Faculty of Science, Charles University,


Šteiger Vladimír, Department of Parasitology, Faculty of Science, Charles University,


Šulcová Tereza, Czech University of Life Sciences, Kamýcká 961/129, 165 00, Praha-

Suchdol, Czechia, [email protected]





















61

Turjanicová Libuše, Department of Parasitology, Faculty of Science, Charles University,


Uhrovič Dalibor, Institute of Parasitology SAS, Hlinkova 3, 040 01 Košice, Slovakia,

[email protected]

Vetešníková Šimková Andrea, Department of Botany and Zoology, Faculty of Science, Ma-

saryk University, Kotlářská 2, 611 37 Brno, Czechia, [email protected]

Vlčková Linda, Department of Parasitology, Faculty of Science, Charles University,


Vokřál Ivan, Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec

Králové, Charles University, Akademika Heyrovského 1203, 500 05, Hradec Králové,

Czechia, [email protected]

Vondráček Oldřich, Department of Parasitology, Faculty of Science, Charles University,


Vorel Jiří, Department of Botany and Zoology, Faculty of Science, Masaryk University,

University campus Bohunice, Kamenice 5, 625 00 Brno, Czechia, [email protected]

Vyčítalová Kateřina, Department of Botany and Zoology, Faculty of Science, Masaryk Uni-


Vyhlídalová Tereza, Institute of Parasitology, Biology Centre, CAS, v.v.i., Branišovská 31,


Total number of registered participants: 62










62

CONFERENCE PARTNERS

Council of Scientific Societies

of Czech Republic

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rvs.avcr.cz

Bioconsult Laboratories

Božejovická 145, 142 01 Praha 4

bioconsult.cz/cs

Eppendorf Czech & Slovakia s.r.o.

Voděradská 2552/16, 215 01 Říčany u Prahy

www.eppendorf.com/CZ-cs

Merck spol. s r.o.

Na Hřebenech II 1718/10, 140 00 Praha

www.merckmillipore.com

Olympus Czech Group, s.r.o.

Evropská 176, 160 41 Praha 6

www.olympus.cz

http://www.bioconsult.cz/cs

http://www.eppendorf.com/CZ-cs

http://www.merckmillipore.com/

https://www.olympus.cz/

63

CONFERENCE PARTNERS

Schoeller Instruments, s.r.o.

Vídeňská 1398/124, 148 00 Praha 4

www.instruments.cz

AnLab, s.r.o.

Vídeňská 1083, 142 20 Praha 4

www.anlab.cz

Carl Zeiss spol. s r.o.

Radlická 14/3201, 150 00 Praha 5

www.zeiss.cz

EDLIN, s.r.o.

Za Kralupkou 440, 277 11 Libiš

www.edlin.cz

Časopis ŽIVA

Vodičkova 40, 110 00 Praha 1

ziva.avcr.cz

http://www.instruments.cz/

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http://ziva.avcr.cz/

64

NOTES

65

NOTES

24th Helminthological Days 2018, Programme &Abstracts

Edited by: Jana Bulantová, Tomáš Macháček

Published by: Charles University, Faculty of Science

Printed by: Tisk Pětka s.r.o., Horova 1631, 252 63 Roztoky u Prahy

1st edition, 2018

Number of copies: 70

© Czech Society for Parasitology, 2018

Cover illustration © Lucie Jedličková, 2018

ISBN 978-80-7444-057-1

24TH HELMINTHOLOGICAL DAYS 2018 abstract book... · 2018. 10. 11. · 24th Helminthological Days...

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