(1) Hd 2014 Sbornik Final

MASARYK UNIVERSITY

21ST HELMINTHOLOGICAL DAYS 2014

Programme & Abstracts

Editors: Jan Brabec, Miroslava Soldánová & Iveta Hodová

BRNO 2014

Conference organizers: Biology Centre of the ASCR, Institute of Parasitology, České Budějovice, Czech Republic Czech Society for Parasitology Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno

Place and date of conference: 21st Helminthological Days, Recreational facility FTVS UK, Dvorce near Stráž nad Nežárkou, Czech Republic, 5–9 May 2014

Programme

ST PROGRAMME OF THE 21 HELMINTHOLOGICAL DAYS

MONDAY, MAY 5

15.00–18.00 Arrival and registration of participants 18.00–18.30 Dinner 19.00–00.00 Get-together-evening

TUESDAY, MAY 6

08.00–09.00 Breakfast

09.15–09.30 Opening ceremony

09.30–10.00 Invited talk: R. Kuchta, T. Scholz: (Self-) infections with helminths.

Session I – Genetics of helminths (Chairman: Martin Kašný) 10.00–10.15 M. Špakulová, M. Bombarová, B. Koubková: Dancing chromosomes: karyotype variation in Paradiplozoon homoion (Monogenea, Diplozoidae). 10.15–10.30 E. Šrámová, D. Jirsová, M. Gelnar, M. Kašný: AFLP: study of genetic variability in Trichinella populations. PhD

10.30–11.00 Coffee break

Session II – Helminthology in genomic era (Chairman: Martin Kašný) 11.00–11.15 J. Bulantová, M. Kašný, D. Potěšil, Z. Zdráhal, L. Staněk, P. Horák: Molecular microscope PART II - “The journey to the inside of the worms”. 11.15–11.30 R. Leontovyč, P. Horák, M. Kašný: The peptidases from Fascioloides magna adults involved in blood digestion. PhD 11.30–11.45 J. Ilgová, B. Koudela, M. Gelnar, M. Kašný: Cysteine peptidase inhibitors of helminths. PhD 11.45–12.00 J. Brabec: Employing the next generation sequencing approach in taxonomical studies of nonmodel parasitic flatworms.

12.00–12.30 Lunch

Session III – Morphology-based studies of helminths (Chairman: Roman Kuchta) 13.30–13.45 F.M. Vieira, T. Scholz, J.L. Luque: Nematodes of the genus Cucullanus Müller, 1777 in the Neotropics: problems and perspectives. 13.45–14.00 F.B. Pereira, J.L. Luque: Are the lizards from Brazil “good friends” of nematode taxonomists? An answer based upon three personal cases. PhD 14.00–14.15 K. Francová, E. Řehulková: Monogeneans of catfish species Schilbe uranoscopus from Kenya. 14.15–14.30 J. Pankrác, P. Horák, M. Kašný: Organogenesis of redia and cercaria of Fascioloides magna (Digenea). BSc/MSc

14.30–15.00 Coffee break

Programme

Session IV – Parasitic flatworm taxonomy (Chairman: Miroslava Soldánová) 15.00–15.15 I. Pokorná, R. Kuchta: Unique diphyllobothriidean tapeworms from reptiles. BSc/MSc 15.15–15.30 P.V. Alves, J.N. Borges, C.P. Santos, J.L. Luque: A redescription of Lobatostoma kemostoma (MacCallum & MacCallum, 1913) (Trematoda: Aspidogastrea) from the Florida pompano fish Trachinotus carolinus (Linnaeus, 1766) off the Brazilian coast. PhD 15.30–15.45 M. Oros, T. Scholz, E. Bazsalovicsová, J. Brabec: Cryptic species diversity in Paracaryophyllaeus (Cestoda: Caryophyllidea), parasites of loaches (Cobitidae) in Eurasia 15.45–16.00 D. Barčák, M. Oros, V. Hanzelová, T. Scholz: Phenotypic plasticity in Caryophyllaeus brachycollis Janiszewska, 1953 (Cestoda: Caryophyllidea), a parasite of cyprinid fishes. PhD

Poster session (5 min presentations) (Chairman: Miroslava Soldánová) 16.00–16.05 M. Chmelíková, B. Koubková, M. Gelnar, M. Kašný: The carp immune response to Eudiplozoon nipponicum. 16.05–16.10 V. Michálková, Š. Mašová, P. Jurajda: Diversity of fish parasites in Prince Gustav Channel (Weddell Sea), Antarctica – expedition 2014: preliminary results. 16.10–16.15 M.L. Kičinjaová, E. Řehulková: Dactylogyrids (Monogenea) parasitizing African tetras (Characiformes: Alestidae) in Zimbabwe. 16.15–16.20 P. Roudnický, J. Ilgová, M. Gelnar, M. Kašný: The protein inhibitors of peptidases from Eudiplozoon nipponicum and their preparation in recombinant form by using the cell-free expression systems. 16.20–16.25 J. Vorel, H. Dvořáková, L. Jedličková, L. Mikeš, M. Gelnar, M. Kašný: The bioinformatic analyses of "-omes" of helminths with special focus on serine peptidases of Eudiplozoon nipponicum.

16.25–17.00 Individual poster session discussion

17.00–17.45 Presentations of sponsors

18.00–18.30 Dinner

20.00–00.00 Campfire night

WEDNESDAY, MAY 7

08.00–09.00 Breakfast

09.30–10.00 Invited talk: F. Stejskal: Imported systemic helminthic infections

Session V – Excretory-secretory products, part 1 (Chairman: Vladimír Skála) 10.00–10.15 L. Jedličková, H. Dvořáková, M. Kašný, L. Mikeš: Digestive peptidases of monogeneans of the family Diplozoidae. PhD 10.15–10.30 L. Ulrychová, A. Leontovyč, M. Horn, J. Dvořák: Characterization of Schistosoma mansoni serine protease 2 (SmSP2). PhD

Programme

10.30–11.00 Coffee break

Session VI – Excretory-secretory products, part 2 (Chairman: Vladimír Skála) 11.00–11.15 R. Šašková, R. Leontovyč, P. Horák, M. Kašný: The expression profile of cathepsin L in developmental stages of Fascioloides magna. BSc/MSc 11.15–11.30 T. Perháčová, L. Mikeš: Isoforms of a cathepsin L-like peptidase in cercariae of Diplostomum pseudospathaceum. BSc/MSc 11.30–11.45 H. Dvořáková, C.R. Caffrey, A.J. O’Donoghue, L. Mikeš: Substrate specificity of recombinant isoforms of cathepsin B1 from the bird schistosome Trichobilharzia regenti. PhD 11.45–12.00 L. Škorpíková, E. Šrámová, J. Ilgová, B. Koudela, M. Gelnar, M. Kašný: Cystatins of the parasitic nematodes from the genus Trichinella. BSc/MSc

12.00–12.30 Lunch

Session VII – Immunology of Trichobilharzia infections (Chairman: Libor Mikeš) 13.30–13.45 K. Chalachanová, L. Lichtenbergová: Immune response of mice on neuroinfection with bird schistosome Trichobilharzia regenti. BSc/MSc 13.45–14.00 T. Macháček, L. Lichtenbergová, J. Bulantová, P. Horák: The role of microglia in the immune response of mice infected by the neurotropic fluke Trichobilharzia regenti. BSc/MSc 14.00–14.15 V. Skála, Z. Jindrová, M. Vostrý, A.J. Walker, P. Horák: Effects of infection with Trichobilharzia regenti on haemocyte activities of the host snail Radix lagotis. PhD 14.15–14.30 M. Vaščiková, P. Horák: Molecular and serological diagnosis of infections caused by Trichobilharzia. BSc/MSc

14.30–15.00 Coffee break

Session VIII – Immunology of host-parasite interactions (Chairman: Andrea Vetešníková Šimková) 15.00–15.15 T. Pakosta, A. Vetešníková Šimková: The immunity and parasitism in gibel carp (Carassius gibelio), a species with dual form of reproduction. PhD 15.15–15.30 A. Cortés, C. Muñoz-Antolí, J. Sotillo, J.G. Esteban, R. Toledo: Intestinal IFN- γ production is associated with protection from clinical signs, but not with elimination of worms, in Echinostoma caproni-infected mice. PhD 15.30–15.45 K. Skipalová, L. Mikeš: Anticoagulation factors and blood uptake by monogeneans of the family Diplozoidae. BSc/MSc 15.45–16.00 L. Turjanicová, M. Kašný, L. Mikeš: Antigens of bird schistosomes of the genus Trichobilharzia: Preparation of recombinant proteins and duck antibody reactivity. PhD

18.00–18.30 Dinner

19.00–00.00 Free evening

Programme

THURSDAY, MAY 8

08.00–09.00 Breakfast

Session IX – Host-parasite interactions (Chairman: Christian Selbach) 09.15–09.30 L. Gettová, A. Gilles, A. Vetešníková Šimková: Dactylogyrus communities in the Barbus hybrid zone of southern France. PhD 09.30–09.45 V. Krasnovyd, A. Vetešníková Šimková: Effect of maternal origin on monogenean infection in fish hybrids. PhD 09.45–10.00 K. Melounová, J. Vadlejch, J. Bulantová, P. Horák, M. Kašný: Interaction of Fascoiloides magna with definitive host. BSc/MSc 10.00–10.15 V. Michálková, M. Ondračková: Conspicuous behaviour of naturally Apatemon-infected killifish Nothobranchius furzeri – a case of parasite manipulation? PhD 10.15–10.30 K. O’Dwyer, R. Poulin: Trematode infection affects intertidal snail attachment strength. PhD

10.30–11.00 Coffee break

Session X – Molecular taxonomy (Chairman: Jan Brabec) 11.00–11.15 J. Zikmundová, S. Georgieva, A. Faltýnková, M. Soldánová, A. Kostadinova: Molecular and morphological characteristion of cryptic species of larval plagiorchiidean trematodes. BSc/MSc 11.15–11.30 S. Georgieva, S. Dallarés, A. Pérez-del-Olmo, M. Carrassón: Hidden in the deep-sea: diversity of Lepidapedon (Digenea: Lepidapedidae) in the Western Mediterranean. PhD 11.30–11.45 C. Selbach, M. Soldánová, S. Georgieva, A. Kostadinova, B. Sures: Show me your spines – Diplostomum spp. diversity in the Ruhr river in Germany. PhD

12.00–12.30 Lunch

15.00–17.00 Stráž nad Nežárkou castle tour (departure at 14:30)

17.30–00.00 Closing ceremony, party with live music at Stráž nad Nežárkou

FRIDAY, MAY 9

08.00–09.00 Breakfast 09.00–11.00 Departure of participants

Abstracts

ABSTRACTS

Abstracts

A REDESCRIPTION OF LOBATOSTOMA KEMOSTOMA (MACCALLUM & MACCALLUM, 1913) (TREMATODA: ASPIDOGASTREA) FROM THE FLORIDA POMPANO FISH TRACHINOTUS CAROLINUS (LINNAEUS, 1766) OFF THE BRAZILIAN COAST

1 2 2 3 P.V. ALVES , J.N. BORGES , C.P. SANTOS AND J.L. LUQUE

1Curso de Pós-Graduação em Biologia Animal, Universidade Federal Rural do Rio de Janeiro, BR 465, Km 7, Seropédica, RJ, 23890-000, Brazil 2Laboratório de Avaliação e Promoção da Saúde Ambiental, Instituto Oswaldo Cruz, Av. Brasil, 4365, Manguinhos, Rio de Janeiro, RJ, Brazil 3Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, CP 74.508, Seropédica, RJ, 23851-970, Brazil

During a survey of fish parasites off the Brazilian coast, adult specimens of Lobatostoma kemostoma were recovered from the intestine of Trachinotus carolinus. Therefore, L. kemostoma is redescribed on the basis of examination of type series and newly collected specimens. A lectotype was also designated. Thirty-eight specimens of T. carolinus were caught by commercial trawlers in coastal zone of Rio de Janeiro State, Brazil. Sampling was carried out from June 2012 to March 2013. Trematodes recovered were fixed and stored in 70% cold alcohol or 5% folmaldehyde solution for molecular and morphological analysis, respectively. Lobatostoma kemostoma can be differentiated from the all congeners by the hindbody (tail) longer than ventral disc and the cephalic lobes with the same shape and size. The highest Blast score between the sequences was shown by Lobatostoma manteri (90% of similarity). The phylogenetic tree from the Maximum Likelihood analysis shows the present sequences of L. kemostoma as sister-group of A. conchicola, while the two sequences of the congeneric species L. manteri clustered together and were close related with L. kemostoma and A. conchicola. Multicalyx elegans which belongs to Multicalycidae presented an intermediate position between members of Aspidogastridae.

Abstracts

PHENOTYPIC PLASTICITY IN CARYOPHYLLAEUS BRACHYCOLLIS JANISZEWSKA, 1953 (CESTODA: CARYOPHYLLIDEA), A PARASITE OF CYPRINID FISHES

1 1 1 2 D. BARČÁK , M. OROS , V. HANZELOVÁ AND T. SCHOLZ

1Institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 04001 Košice, Slovakia 2Institute of Parasitology, Biology Centre of the Academy of Science of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic

Recent molecular phylogenetic studies on fish tapeworms of the genus Caryophyllaeus Gmelin, 1790 (Cestoda: Caryophyllidea), parasites of cyprinid fishes in the Palaearctic Region, has revealed unexpected phenotypic plasticity that seems to be driven by definitive hosts. In the present study, Caryophyllaeus brachycollis Janiszewska, 1953 is redescribed and its two morphotypes are circumscribed on the basis of newly collected specimens. Morphotype 1 from barbels (Barbus spp. including the type-host Barbus barbus (L.); Barbinae) and chubs (Squalius spp.; Leuciscinae) is characterised by a more robust body with the spatulate scolex, which is only slightly wider than a very short neck region, and the anterior position of the testes and vitelline follicles, which begin immediately posterior to the scolex. Specimens of morphotype 2 from breams (Abramis, Ballerus and Blicca; Abraminae), which were previously misidentifed as Caryophyllaeus laticeps (Pallas, 1781), possess a more slender body with a flabellate scolex, which is much wider than a long neck, and the first testes begin at a considerable distance posterior to the first vitelline follicles. Despite conspicuous differences in the scolex morphology and the anterior extent of the testes and vitelline follicles, both morphotypes are identical in the morphology of the posterior end of the body, in particular that of the cirrus-sac, which is large, thick-walled, elongate pyriform, and contains a long cirrus, and in the distribution of the vitelline follicles, which surround medially vas deferens near the cirrus-sac. The presence of phenotypic plasticity in morphological characteristics previously used in differentiation of species of Caryophyllaeus may confound species identification, which is crucial for any biodiversity, ecological and evolutionary studies. To avoid these potential problems, combination of morphological and molecular data is strongly recommended.

Acknowledgement: Slovak Research and Development Agency (Project No. APVV-0653-11), Grant Agency VEGA (No. 2/0129/12), Czech Science Foundation (Project No. P505/12/G112) and Institute of Parasitology (RVO: 60077344).

Abstracts

EMPLOYING THE NEXT GENERATION SEQUENCING APPROACH IN TAXONOMICAL STUDIES OF NONMODEL PARASITIC FLATWORMS

J. BRABEC

Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic

Next Generation Sequencing techniques represent a rapidly developing field whose recent technical advances turned it into an affordable source of unprecedented amounts of genomic data that can be exploited even in studies of nonmodel organisms. Here we have employed a shotgun genome sequencing approach using Illumina MiSeq platform to allow characterization of several genome-wide molecular markers to study intraspecific and population genetics level aspects of two flatworm parasite taxa, trematode Diplostomum sp. and cestode Bothriocephalus acheilognathi. Rather than optimizing the parasite genomic DNA to increase data yields for a given specific purpose prior to Illumina sequencing (e.g. enriching for mitochondrial DNA for mitochondrial genome reconstruction, enriching for repetitive regions for microsatellite marker development), we ran two Illumina MiSeq runs on untreated genomic DNAs isolated by routinely used genomic DNA isolations kits and compared the outputs by (i) assessing the coverage of mapped mitochondrial genomic reads on de novo assembled mitochondrial genomes; and (ii) screening for potentially amplifiable microsatellite loci in each of the four (Diplostomum) and six (Bothriocephalus) genomic libraries prepared.

Acknowledgement: Project Postdok_BIOGLOBE (CZ.1.07/2.3.00/30.0032) co-financed by the European Social Fund and the state budget of the Czech Republic.

Abstracts

MOLECULAR MICROSCOPE PART II - “THE JOURNEY TO THE INSIDE OF THE WORMS”

1 1,2 3 3 4 1 J. BULANTOVÁ , M. KAŠNÝ , D. POTĚŠIL , Z. ZDRÁHAL , L. STANĚK AND P. HORÁK

1Department of Parasitology, Faculty of Science, Charles University in Prague, Vinicna 7, 128 00 Prague, Czech Republic 2Department of Botany and Zoology, Faculty of Science, Masaryk University Brno, Kamenice 753/5 625 00 Brno Bohunice, Czech Republic 3National Centre for Biomolecular Research, Masaryk University Brno, Kamenice 753/5, 625 00 Brno Bohunice, Czech Republic 4Institute of Pathology of 1st Faculty of Medicine and General Teaching Hospital, Charles University Prague, Studničkova 2, 128 00 Prague, Czech Republic

Currently, in the era of "Next Generation Sequencing", the genome/proteome databases are still faster and faster filled with comprehensive sequential data. There are many possibilities how to effectively use these robust data sets, e.g. for interpretation of mass spectra of analyzed proteins leading to their identification, prediction of protein functions, or better understanding of protein specific roles in biological systems. In the context of protein functions it is useful to complete this computational information by localization of protein specific expression in cells/tissues and protein mode of action. Localization of the selected protein in particular tissues by using common immuno- techniques is a time consuming process that usually includes cloning of gene, preparation of the recombinant form, its purification, immunization of experimental animals, purification of antibodies, preparation of histological slides and immunohistochemical staining by use of sera from immunized animals. Several years ago (Helminthological days 2008, Ředkovec), the potential of mass spectrometry in modern proteomic studies focused on parasites, namely trematodes, was presented as a possible “shortcut” in the process of protein immuno-localization mentioned above. MALDI-MSI technique (Matrix-Assisted Laser Desorption/Ionization Mass Spectrometric Imaging) was highlighted as a perspective method for direct determination and monitoring of biomolecules in the organic tissue. Recently we optimized the method also for identification and localization of particular proteins; we adopted microdissection technique combined with mass spectrometry analysis. Predefined organs (e.g. intestine, penetration glands, vitellaria) or specific tissue parts (tegument, musculature) were thoroughly microdissected from cryosections of two model species - Trichobilharzia regenti (cercariae) and Fascioloides magna (adults). Microdissection ensures there subsequent precise analysis of protein distribution directly in organs of interest, without possible contamination. By application of this approach cathepsin L from F. magna intestine was identified.

Acknowledgement: Czech Science Foundation (Project No. 13-29577S) and Charles University in Prague (UNCE 204017, PRVOUK P41, SVV 267210/2013, GAUK 502313).

Abstracts

INTESTINAL IFN-Γ PRODUCTION IS ASSOCIATED WITH PROTECTION FROM CLINICAL SIGNS, BUT NOT WITH ELIMINATION OF WORMS, IN ECHINOSTOMA CAPRONI-INFECTED MICE

1 1 1,2 1 1 A. CORTÉS , C. MUÑOZ-ANTOLÍ , J. SOTILLO , J.G. ESTEBAN AND R. TOLEDO

1Departament de Biologia Cel·lular i Parasitologia, Facultat de Farmàcia, Universitat de València. Avda. Vicent Andrés Estellés s/n, 46100 Burjassot, Valencia, Spain 2Center for Biodiscovery and Molecular Development of Therapeutics, James Cook University, Cairns, Queensland, Australia

Echinostoma caproni (Trematoda: Echinostomatidae) is an intestinal trematode, with no tissue phases in the definitive host. Although E. caproni is able to parasitize a wide range of laboratory rodent hosts its compatibility differs considerably among rodent species. In high compatible hosts, such as ICR mice, the infection becomes chronic, while in hosts of low compatibility, such as rats, the worms are expelled a few weeks post-infection (wpi). Because of this characteristic, the E. caproni/rodent model has been extensively used to study several aspects of the host-parasite interactions. In the present study we have investigated the course of the infection in three mouse strains: ICR (a host of high compatibility with E. caproni), BALB/c (a prototypical Th2 strain), and BALB/c deficient for IFN-γ (IFN-γ -/-). For this purpose the local immune response generated in each host strain was characterized by analyzing the cytokine expression profile at the ileal mucosa. As previously described, in ICR mice the infection was characterized by the elevated expression of IFN-γ and iNOS in the intestine, concomitantly with the lack of clinical signs. In contrast, the infection was more virulent in BALB/c and IFN-γ -/- mice, which developed a severe form of the disease, coinciding with the absence of local IFN-γ and iNOS expression. Both two strains showed loss of weight, hackled hair, diarrhea, perianal irritation and inflammation and they looked emaciated from 2-3 wpi. At the gross examination of the intestines, intense emaciation was also appreciated, with extremely thin intestinal walls. Nonetheless, these signs were more pronounced in IFN-γ deficient mice, in which the disease was lethal in the first weeks of the infection. At tissue level, infected mice of the three strains showed erosion at the villi tip and crypt hyperplasia, although increases in the villi lenght were not found in IFN-γ deficient mice, as they were in ICR and BALB/c strains. The analysis of different parameters of the infection in each host strain, such as goblet and mast cell populations or patterns of mucins expression, which are known to play a protective role against the virulence induced by the parasite, revealed that most of the features were similar in the three mouse strains. This suggests that IFN-γ, and probably iNOS, may play a major role in the protection against severe disease. Although these mediators do not contribute to parasite clearance, since no significant differences were detected in the percentage of worms recovered in each case, they may protect against the injurious effects of the parasite on the host, preventing the development of morbidity and lethality. These results also indicate that the development of inflammatory responses in helminth infections is not necessarily associated with severe forms of disease.

Abstracts

SUBSTRATE SPECIFICITY OF RECOMBINANT ISOFORMS OF CATHEPSIN B1 FROM THE BIRD SCHISTOSOME TRICHOBILHARZIA REGENTI

1 2 3 1 H. DVOŘÁKOVÁ , C.R. CAFFREY , A.J. O'DONOGHUE AND L. MIKEŠ

1Department of Parasitology, Faculty of Science, Charles University of Prague, Viničná 7, 128 44 Prague 2, Czech Republic 2California Institute for Quantitative Biosciences, University of California San Francisco, Byers Hall BH501E, 1700 4th Street, San Francisco, CA 94158-2330, USA 3Department of Pharmaceutical Chemistry, University of California San Francisco, Genentech Hall MC 2280600, 16th Street, San Francisco, CA 94158-2330, USA

One of the keys to understanding the biological role of peptidases in any organism is the knowledge of a preferred cleavage site within their substrates (substrate specificity of peptidases). The information about specificity can provide a clue to natural substrates of a particular protease and may serve as a basis for efficient substrate or inhibitor design. Therefore, two recombinant isoforms of cathepsin B1 of the bird schistosome T. regenti (TrcB1.1 and TrCB1.4, GenBank AY648119 and -22) expressed in Pichia pastoris have been analyzed for substrate specificity using a positional scanning synthetic combinatorial library. This library permits determination of amino acid preferences of peptidases in a defined position within oligopeptidesubstrates (P1-P4). Recombinant TrCB1.1 exhibits preference for Lys and Arg in P1 position, a broader specificity in P2 position (Met>Val>Ala>Ser>nLeu> Ile>Leu>Asp), Leu and other hydrophobic amino acids in P3 and Arg, Lys in P4 position of the oligopeptide substrate. Recombinant TrCB1.4 has a broader specificity in P1 position (Arg> Met>nLeu>Lys), but has a narrow specificity in P2 position (mainly small aliphatic hydrophobic amino acids). In the case of P2, both recombinant enzymes differ from most of cathepsins B, which generally prefer large hydrophobic amino acids in this position. Moreover, recombinant TrCB1.4 was inactive towards Arg in P2. Cathepsins B of most organisms (including TrCB1.1) are able to accept Arg in P2 of the substrate. There are no significant differences in preferences between both recombinant isoforms in P3 and P4 positions of the peptide substrate. Although the amino acid sequence identity of TrCB1 isoforms is high (98-99% between the putatively active isoforms), even small changes in amino acid sequences may cause variations in substrate specificities. The consequences of different amino acid preferences of the two recombinant isoforms will be discussed in the context of their putative biological functions.

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

Abstracts

MONOGENEANS OF CATFISH SPECIES SCHILBE URANOSCOPUS FROM KENYA

K. FRANCOVÁ AND E. ŘEHULKOVÁ

Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic

Catfishes (order Siluriformes) represent a diverse group of ray-finned fish (Actinopterygii), including 37 families and living mostly in tropical South America, North America, Africa and Asia. In African countries, 66 species (11 genera) of monogeneans have been recorded to parasitize catfishes (species belonging to 10 genera examined). From Kenya, no records about monogeneans in catfishes existed until research were carried out by our team. Fish were sampled in Turkana Lake in 2008 and 2009 and 12 siluriform species were examined for parasites. Seven siluriform species (belonging to genera Auchenoglanis, Chrysichthys, Bagrus, Clarias, Heterobranchus and Schilbe) were found to be infected with monogeneans (10 species belonging to genera Bagrobdella, Protoancylodiscoides, Quadriacanthus, Schilbetrema and Schilbetrematoides). Majority of monogenean species recorded probably represent species new for science. This is particularly true for monogenean species found in butter catfish Schilbe uranoscopus (first records of monogenean infection in this fish species). S. uranoscopus, in comparison with the other host species collected, was found to be infected with the highest number (4) of monogenean species: three Schilbetrema species and one Schilbetrematoides species, on which we are currently focused.

Acknowledgements: ECIP – Centre of excellence, Czech Science Foundation (Project No. P505/12/G112)

Abstracts

HIDDEN IN THE DEEP-SEA: DIVERSITY OF LEPIDAPEDON (DIGENEA: LEPIDAPEDIDAE) IN THE WESTERN MEDITERRANEAN

1,2 3 3,4 3 S. GEORGIEVA , S. DALLARÉS , A. PÉREZ-DEL-OLMO AND M. CARRASSÓN

1Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic 2 Faculty of Science, University of South Bohemia, Branišovská 31, 370 05 Ceske Budejovice, Czech Republic 3Departament de Biologia Animal, Biologia Vegetal i Ecologia, Universitat Autònoma de Barcelona, Cerdanyola, 08193 Barcelona, Spain 4Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Universitat de València, PO Box 22085, 46071 Valencia, Spain

Deep-sea parasite faunas are considered much poorer than costal even though they still have not been enough studied. The genus Lepidapedon Stafford, 1904, comprised of parasites of deep-sea or cold shallow-water marine teleosts, represents the most speciose and probably the most dominant digenean genus in the bathypelagic zone. However, the geographical distributional range of Lepidapedon spp. is rather limited and they are predominantly reported from the North Atlantic and Northeast Pacific. Numerous specimens of Lepidapedon were obtained from five deep-sea gadiform fish species (Coelorinchus labiatus, Mora moro, Lepidion lepidion, Phycis blennoides and Trachyrinchus scabrus) collected at depths between 558 m and 1754 m in four locations in the Western Mediterranean. Representative samples were characterised both morphologically and molecularly. The molecular data (nad1 gene and the lsrDNA) were used to aid identification and infer phylogenetic relationships among the members of the genus. Concatenated sequences from both loci and nad1 gene on its own were used for constructing phylogenetic hypotheses. Morphological and molecular analyses revealed that all collected specimens from the four host species belong to two species: Lepidapeon desclersae Bray & Gibson, 1995 and L. guevarai Lopez-Roman & Maillard, 1973. This first study on Lepidapedon spp. from the Mediterranean deep-sea provides for the first time molecular data for both species from their type-hosts; both species exhibited high haplotype diversity (h=0.924) and low host specificity.

Acknowledgements: Spanish Ministry of Science and Innovation (MICINN) project ANTROMARE (CTM2009-12214-C02-02) and ECIP – Centre of excellence, Czech Science Foundation (Project No. P505/12/G112).

Abstracts

DACTYLOGYRUS COMMUNITIES IN THE BARBUS HYBRID ZONE OF SOUTHERN FRANCE

1 2 1 L. GETTOVÁ , A. GILLES AND A. ŠIMKOVÁ

1Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, Brno, 611 37, Czech Republic 2Aix-Marseille Université, IMBE, UMR CNRS 7263, Evolution Génome Environnement, Case 36, 3 Place Victor Hugo, 13331 Marseille Cedex 3, France

The genus Dactylogyrus represents the species richest group within the family Dactylogyridae (Monogenea). In our study, we aimed to study the composition of Dactylogyrus communities in two cyprinid fish species, Barbus meridionalis and B. barbus, forming the hybrid zone in southern France. Based on previously published data from this area, B. barbus is parasitized specifically by Dactylogyrus malleus, while D. carpathicus and D. dyki have been reported for both Barbus species under investigation. Since the host distribution range was shown to affect the level of parasite infection, we hypothesized higher parasite load in populations of widely distributed B. barbus comparing with geographically restricted B. meridionalis populations. Furthermore, we assumed lower hybrid susceptibility to Dactylogyrus parasites as a result of so called “hybrid vigor”. Over the period 2007-2013, 327 fish were dissected for metazoan parasites in both sympatric and allopatric Barbus populations. Fish determination was performed using 30 microsatellite markers and revealed the occurrence of 40 hybrid individuals in seven Barbus populations. Of all parasites, Dactylogyrus species represent the most dominant group in almost all studied populations. D. carpathicus parasitized both Barbus species, while D. malleus was specific for B. barbus. These parasites reached the highest values of abundance and prevalence in allopatric B. barbus populations. D. dyki was present solely in allopatric population of B. meridionalis that is in disagreement with previously published data. Furthermore, D. extensus (specific to Cyprinus carpio) represented a rare species firstly detected in B. meridionalis individuals. Overall, significant trend towards higher abundance of Dactylogyrus parasites in B. barbus compared to both B. meridionalis and hybrids was detected. Further extensive sampling of sympatric populations is required to investigate the composition of Dactylogyrus communities along the Barbus hybrid zone.

Acknowledgement: Czech Science Foundation (Project No. P505/12/0375).

Abstracts

IMMUNE RESPONSE OF MICE ON NEUROINFECTION WITH BIRD SCHISTOSOME TRICHOBILHARZIA REGENTI

K. CHALACHANOVÁ AND L. LICHTENBERGOVÁ

Department of Parasitology, Charles University, Viničná 7, 12844 Prague 2, Czech Republic

Larvae of neurotropic avian schistosome T. regenti are able to penetrate the skin of mammals and migrate via peripheral nerves to the central nervous system (CNS). In the mammalian host, the parasites never develop to adult flukes. They are eliminated by host immune system. Mechanism of this immune reaction is not completely described. Previous studies on mice infected with T. regenti showed, that immune cells produce a mixed Th1/Th2 cytokine response after primo-infection. Th17 cytokine response is expected. Th2 immune response is dominating after repeated infections. Aim of our study is to measure intracellular cytokines in peripheral lymphocytes of mice experimentally infected with T. regenti and to observe polarization (Th1/Th2/Th17) of the immune response after primo- infection and during repeated infections. Astrocytes are active players in the immune response against infection of the CNS. Activated astrocytes regulate the immune response by production of different cytokines and mediators of inflammation such as nitric oxide. It is our intention to elucidate the role of the astrocytes during T. regenti neuroinfection. For this purpose, primary cultures of astrocytes from newborn mice are prepared. Subsequently, the astrocytes are stimulated with homogenate of in vitro transformed cercariae and the levels of selected cytokines (IL-1, IL-6, TNF-α) and nitric oxide are measured.

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

Abstracts

THE CARP IMMUNE RESPONSE TO EUDIPLOZOON NIPPONICUM

1 1 1 1,2 M. CHMELÍKOVÁ , B. KOUBKOVÁ , M. GELNAR AND M.KAŠNÝ

The fish immune system as well as the immune system of other vertebrates consists of cellular or humoral specific and nonspecific factors. Important cellular elements of the Common carp (Cyprinus carpio) are the leucocytes, the mucous cells and the epithelial cells. Main components of fish specific humoral immunity are antibodies, five types of immunoglobulins (Ig) – IgM, IgD, IgZ, IgT and IgH. During our research work the blood samples (sera) will be collected from carps parasitized by E. nipponicum and from non-infected ones. The excretory-secretory products and whole worm extracts will be purified and used as antigens in immuno-chemical methods. Similarly the parasite samples will be also incubated with commercially available polyclonal Anti-Carp IgM antibodies with the aim to record the significant reaction by using ELISA and Western blot. The most antigenic protein molecules will be identified by mass spectrometric methods.

Acknowledgements: Czech Science Foundation (Project Nos. P506/12/1258, GBP505/12/G112) and Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013).

Abstracts

CYSTEINE PEPTIDASE INHIBITORS OF HELMINTHS

1 3 1 1, 2 J. ILGOVÁ , B. KOUDELA , M. GELNAR AND M. KAŠNÝ

1Department of Botany and Zoology, Faculty of Science, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic 2Department of Parasitology, Faculty of Science, Charles University in Prague, Viničná 7, 128 44 Prague 2, Czech Republic 3Department of Pathological Morphology and Parasitology, Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Science Brno, Palackého 1/3, 612 42 Brno, Czech Republic

Investigating the functions of various bioactive molecules produced by parasites expands our understanding of the principles of host-parasite interaction. Several of these compounds were proven to play a key role in a manipulation of the host immune system and thus maximizing the parasite success; among them also the cysteine peptidase inhibitors (CPI) were recorded. Our work is focused on characterization of the properties of CPI expressed by four helminth model species representing four big taxonomic groups – Monogenea, Cestoda, Trematoda and Nematoda. Prior to the functional analysis we searched the transcriptomic and/or genomic data of Eudiplozoon nipponicum, Hymenolepis diminuta, Fascioloides magna and Trichinella spiralis for the presence of conserved cystatin domains. We identified distinct families of CPI (stefins, cystatins and kininogens) and observed differences among the studied species. We designed specific/degenerate primers; amplified, cloned and sequenced the selected genes. The recombinant forms of particular CPI will be prepared and characterized in detail to reveal their function.

Acknowledgements: Masaryk University (Project No. MUNI/A/0888/2013), Czech Science Foundation (Project Nos. GBP505/12/G112, GAP506/12/1258), Ministry of Education, Youth and Sport Czech Republic (Project CONTACT II No. LH12096) and Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013).

Abstracts

DIGESTIVE PEPTIDASES OF MONOGENEANS OF THE FAMILY DIPLOZOIDAE

1 1 1,2 1 L. JEDLIČKOVÁ , H. DVOŘÁKOVÁ , M. KAŠNÝ AND L. MIKEŠ

1Department of Parasitology, Faculty of Science, Charles University in Prague, Viničná 7, 128 44 Prague 2, Czech Republic 2Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic

Members of the family Diplozoidae are blood-feeding monogeneans mainly parasitizing on the gills of cyprinid fishes. They may have pathogenic effects on their hosts caused by mechanical damage of gill filaments accompanied by secondary infections, or by induction of hypochromic mycrocytic anemia. The information about blood processing mechanisms in monogeneans of the subclass Heteronchoinea (Polyopisthocotylea) stems from ultrastructural or histochemical analyses. It has been assumed that digestion is intracellular and takes place in a specialized type of gut cells. Only little information exists about the biochemistry of digestion. Therefore we focused on biochemical and molecular characterization of peptidases, which can be involved in digestion of blood proteins. The presence of the cysteine class of peptidases, mainly cathepsin L, was approved in samples of excretory/secretory products and in soluble protein extracts from adult Paradiplozoon bliccae and Eudiplozoon nipponicum by using fluorogenic substrates, specific inhibitors and a biotinylated/fluorochrome-labelled affinity probe DCG-04. In the gels after electrophoresis or on blots we observed labelled bands of 35 kDa in the case of both species and 24 kDa for E. nipponicum only. Soluble protein extracts of worms were separated by 2D electrophoresis; detected spots around 35 kDa for P. bliccae and around 35-25 kDa for E. nipponicum were identified by mass spectrometry analyses as cathepsins L. On the basis of transcriptome analyses we have predicted 30 peptidase genes in E. nipponicum, 9 of which may be involved in nutrient digestion. Cathepsin L and cathepsin B of E. nipponicum have been expressed in Pichia pastoris yeast and are being biochemically characterized.

Acknowledgements: Czech Science Foundation (Project No. P506/12/1258), Charles University in Prague (Project No. 502313).

Abstracts

DACTYLOGYRIDS (MONOGENEA) PARASITIZING AFRICAN TETRAS (CHARACIFORMES: ALESTIDAE) IN ZIMBABWE

M.L. KIČINJAOVÁ AND E. ŘEHULKOVÁ

Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno

The Alestidae, often called “African tetras”, is the most speciose (with about 120 species) family of characiform fishes in Africa. These fishes are known to be parasitized by monogeneans representing 3 genera of the Dactylogyridae: Annulotrema, Characidotrema and Afrocleidodiscus. During several field campaigns in Senegal (2004–2008), Kenya (2008– 2009), Sudan (2008) and Zimbabwe (2011–2012), a total of 15 species of the Alestidae have been examined for monogeneans and more than 30 monogenean species (3 genera) of the Dactylogyridae were found. Our current research dealing with the taxonomic evaluation of monogeneans collected from 2 species (2 genera, i.e. Hydrocynus and Brycinus) of the Alestidae from the Kariba Lake and Zambezi River in Zimbabwe revealed the presence of 6 species of Annulotrema and 1 species of Afrocleidodiscus. Three species of them are probably new for science. Morphological analysis of sclerotized structures of the “Kenya” and “Zimbabwe” species of Annulotrema collected by us suggests that there are at least 3 groups within the dactylogyrids parasitizing these fishes, with regard to the basic structure of the male copulatory organ (MCO). The first group includes 5 species with MCOs composed of an arcuate copulatory tube articulated to an accessory piece with lightly sclerotized proximal part; the second group (including 2 species) is characterized by MCO with sock-like base and non-articulated accessory piece with richly intertwined distal part; the third group consists of 3 species with an oval base of the copulatory tube and articulated accessory piece with sickle shaped distal part.

Acknowledgements: Czech Science Foundation (Project Nos. LC 522, P505/12/G112).

Abstracts

EFFECT OF MATERNAL ORIGIN ON MONOGENEAN INFECTION IN FISH HYBRIDS

V. KRASNOVYD AND A. VETEŠNÍKOVÁ ŠIMKOVÁ

Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlařská 2, Brno, 611 37, Czech Republic

The close affinity to a single host species for some parasite species (termed as highly host specific) such are monogeneans is predicted in link with host-parasite coevolution. However, hybridization may interrupt the system of co-adaptation genes. Recently, there is limited information concerning the effect of maternal origin on parasite infection in hybrids. In our study, common bream (Abramis brama), roach (Rutilus rutilus) and their respective hybrids from Hamry reservoir (Czech Republic) were collected in spring and autumn 2011, and investigated for the presence of metazoan parasites. Identification of the fish individuals was performed using morphological characters and molecular diagnostic based on the sequence analysis of mitochondrial DNA – partial cytochrome b gene. The aim of this study was to compare monogenean infection between two parental species and hybrids with different maternal origin. Diversity of monogenean communities was higher but parasite abundance was lower in hybrids where compared to parental species. Regarding maternal origin of hybrids the predominance of individuals with common bream maternal origin was found. The potential effect of maternal origin on parasite infection in hybrids was analyzed.

Acknowledgments: Czech Science Foundation (Project No. P505/12/0375).

Abstracts

(SELF-) INFECTIONS WITH HELMINTHS

R. KUCHTA AND T. SCHOLZ

Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, 370 05 České Budějovice, Czech Republic

The experimental (self-) infections with helminthes can be re-interpreted from a perspective that was usually not on the mind of scientists, who, mostly long time ago, subjected themselves and other individuals to these experiments. In most cases, (self-) infections were used to elucidate life cycles of parasites, their transmission, host specificity and/or pathogenicity, data which were difficult or even impossible to obtain by other means. Interestingly, many of these experiments remain poorly known as they were often published in obscure or non-English journals or have been forgotten. Moreover, some of them are incompatible with current ethical rules. However, with recent research addressing the possible beneficial impact of intestinal helminths on a range of autoimmune diseases in humans, these studies, although often not complying with current ethical rules, provide valuable information. Here, we critically review cases, in which humans were infected with parasites.

Abstracts

THE PEPTIDASES FROM FASCIOLOIDES MAGNA ADULTS INVOLVED IN BLOOD DIGESTION

1 1 1,2 R. LEONTOVYČ , P. HORÁK AND M. KAŠNÝ

The giant liver fluke Fascioloides magna is in Europe and Czech Republic known as introduced species with a high pathogenic impact to the spectrum of its definitive hosts. The pathogenicity is mainly caused due to its size (up to 8 cm), tissue migration and hematophagy. The blood digestion is facilitated by the spectrum of proteolytic enzymes – peptidases. The molecular/biochemical studies shows that the peptidases do not work independently but they act in proteolytic cascades. Recent research on proteolytic enzymes of the Fasciolidae family is mainly focused on molecules with their potential use as a vaccines and drug targets, e.g. Cathepsin L. The functional characteristics of other important hemolytic enzymes are scant. Our work is primarily focused on characterization of five F. magna peptidases, which are potentially involved in the blood digestion – Cathepsins L, D, B, Asparaginyl endopeptidase and Leucyl aminopeptidase. By using the deep bioinformatic analysis of the F. magna transcriptome we recorded full length DNA sequences of all selected peptidase genes. The expression of recombinant FmCB1, FmCB2 and FmAE in Escherichia coli was successful and the expressed proteins were identified by mass spectrometry. The further functional peptidase properties such as substrate specificity, processing and degradation of macromolecular substrates will be investigated.

Acknowledgements: Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013, GAUK 502313).

Abstracts

THE ROLE OF MICROGLIA IN THE IMMUNE RESPONSE OF MICE INFECTED BY THE NEUROTROPIC FLUKE TRICHOBILHARZIA REGENTI

T. MACHÁČEK, L. LICHTENBERGOVÁ, J. BULANTOVÁ AND P. HORÁK

Department of Parasitology, Faculty of Science, Charles University in Prague, Viničná 7, 128 44 Prague 2, Czech Republic

Invasion of the central nervous system (CNS) by parasitic worms is a well known phenomenon. Although originally considered to be a privileged site without immune surveillance, CNS possesses mechanisms, which can be used to fight with invaders, including parasitic helminths. In our research, we focused on the neurotropic dixenic fluke Trichobilharzia regenti. Its larvae (schistosomula) migrate through the spinal cord and the brain of ducks (definitive hosts) and mice (accidental hosts used as our model system). Evaluation of the activation of murine microglia, resident CNS macrophages, during the infection is in progress. Using MHC class II-eGFP knock-in C57BL/6 mice and fluorescently labelled worms we observed MHC class II positive cells in proximity of the parasites in the spinal cord. The real nature of these cells (possibly microglia) has to be elucidated by correlative microscopy. To assess production of nitric oxide (NO) and selected cytokines (IL-1, IL-6, TNF-alpha) by microglia in a response to T. regenti antigens, microglia are obtained from neonatal mouse pups, cultured in vitro and exposed to homogenates of schistosomula. NO and cytokines concentrations are measured by Griess assay and ELISA, respectively. Preliminary data suggest increased production of NO in stimulated microglia, which is to be verified by detection of inducible NO-synthase.

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

Abstracts

INTERACTION OF FASCIOLOIDES MAGNA WITH DEFINITIVE HOST

1 3 1 1 1,2 K. MELOUNOVÁ , J. VADLEJCH , J. BULANTOVÁ , P. HORÁK AND M. KAŠNÝ

1Department of Parasitology, Faculty of Science, Charles University in Prague, Vinicna 7, 128 00 Prague, Czech Republic 2Department of Botany and Zoology, Faculty of Science, Masaryk University Brno, Kamenice 753/5 625 00 Brno Bohunice, Czech Republic 3Department of Zoology and Fisheries, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamycka 957, 165 21 Prague, Czech Republic

Fascioloides magna is a veterinary important trematode parasiting in a wide range of ruminants as definitive hosts. This fluke was introduced to Europe from North America in the 19th century with the herd of deer Wapiti and adapted to European deer. Fascioloides magna can infects primarily cervids and also the farm animals - cattle, horses, sheep and goats. In the body of definitive host the fluke is located in the livers causing a severe damage of this organ, which can lead to death of the host. The definitive hosts can be categorized into three groups: 1. “specific definitive hosts” (e.g. Cervus elaphus, Dama dama, Capreolus capreolus) - the eggs are released via bile ducts from liver pseudocyst to the host intestine; 2. “nonspecific definitive hosts (dead-end hosts)” (Alces alces, Lama glama, Bos taurus) - eggs are trapped in liver pseudocysts not connected to the bile duct system with the host´s intestine; 3. “non-typical hosts (aberrant hosts)” (Ovis aries, Capra hircus, Rattus norvegicus, Oryctolagus cuniculus) - immature flukes continuously migrate throughout the host body. Since we are interested in the host-parasite interaction, we investigated the effects of F. magna infection on the serum biochemical parameters, hematocrit and level of immunoglobulins of six experimentally infected domestic rabbits (Oryctolagus cuniculus f. domesticus, Hyplus hybrid). Twelve weeks after infection the model animals were dissected and one immature fluke was recorded.

Acknowledgements: Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013).

Abstracts

CONSPICUOUS BEHAVIOUR OF NATURALLY APATEMON-INFECTED KILLIFISH NOTHOBRANCHIUS FURZERI – A CASE OF PARASITE MANIPULATION?

1,2 2 V. MICHÁLKOVÁ AND M. ONDRAČKOVÁ

1Department of Botany and Zoology, Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno - Bohunice, Czech Republic 2Institute of Vertebrate Biology, v.v.i., Academy of Sciences of the Czech Republic, Květná 8, 603 65 Brno, Czech Republic

Parasites with complex life cycles may affect their hosts in many ways to increase the probability of transmission to the next host. In this study, we investigated the behaviour of killifish Nothobranchius furzeri naturally infected by larval trematodes Apatemon sp. from Mosambique. Apatemon (Trematoda) is a parasite with complex life cycle that requires the ingestion of the second intermediate host (fish) by the definitive host (piscivorous birds) to complete its life cycle. In the fish host, metacercariae are located in body cavities. In killifish, metacercariae occur exclusively in cerebral cavity which may indicate possible effects on host behaviour. We performed two series of experimental observation to compare behaviour of Apatemon-infected and control killifish: 1) location of the fish in the aquarium without interruption and 2) behavioural responses of killifish to simulated avian predation. Apatemon- infected killifish often remained at the water surface in both series of observation. In the case of simulated attack, the fish displayed conspicuous swimming behaviour manifested by multiple jumps above the water surface, uncoordinated movements and rotation in all directions at the time of the attack and shortly after it. On the contrary Apatemon-uninfected control killifish were located close to the water surface only rarely and, in the observation without interruption, spent most of the time in the lower two-thirds of the aquarium. When simulated the avian attack, the control fish showed an escape response away from the site of the attack into the deeper water or to the walls of the aquarium. Conspicuous behaviour of infected killifish which was even more intense in the case of disturbance may potentially serve as a mechanism to increase the probability of predation by the definitive bird host and thus represent the parasite manipulation. However, this host-parasite system requires a more detailed study to a better understanding of the mechanisms of these host-parasite interactions.

Acknowledgements: Czech Science Foundation (Project Nos. P505/12/G112).

Abstracts

DIVERSITY OF FISH PARASITES IN PRINCE GUSTAV CHANNEL (WEDDELL SEA), ANTARCTICA – EXPEDITION 2014: PRELIMINARY RESULTS

1,2 1 2 V. MICHÁLKOVÁ , Š. MAŠOVÁ AND P. JURAJDA

During the Czech Antarctic expedition 2014 field investigations were carried out to contribute to the knowledge on fish parasites in Antarctica. From January to March 2014 in total 102 specimens of six fish species (Notothenia coriiceps – 14 individuals, Trematomus bernacchi – 30, T. hansoni – 32, T. newnesi – 23, Parachaenichthys charcoti - 2, Pagotenia borshgrewincki – 1 specimen) were examined for parasites. The fish were caught with gill nets and fish rods in the Prince Gustav Channel (depth about 5–25 m) in front of the Johann Gregor Mendel Station on the James Ross Island. Altogether 7740 metazoan parasites (including ectoparasites: Monogenea – 1326, Crustacea – 44, Hirudinea – 20, and endoparasites: Nematoda – 3263, Acanthocephala – 2053, Cestoda – 758 and Trematoda – 276) were found. Also Myxozoa were present in 10 fish individuals. The spatial distribution on fish gills of 1157 monogeneans (family Gyrodactylidae, Dactylogyridae and Capsalidae) was determined. Mean overall parasite abundance across the host species was 76. Maximum abundance (565) was observed in Notothenia coriiceps, which was the most parasitized host species with the mean parasite abundance of 117 and prevalence of 100 %. The lowest parasite abundance (51) was recorded in T. bernacchi. The prevalence in Trematomus spp. was 100 % with the mean abundance 71. All fish were infected with high intensities of nematodes (mean abundance 37, prevalence 97 %). The prevalence of Corynosoma spp. (Acanthocephala) ranged from 39% in T. newnesi to 100% in N. coriiceps. Crustacea and Hirudinea were found rarely and mainly on T. bernacchi and N. coriiceps.

Acknowledgements: Masaryk University and Czech Science Foundation (Project No. P505/12/G112). The authors are grateful to the staff of the Antarctic expedition 2014 in the Czech Antarctic Station ‘‘J. G. Mendel’’ in the James Ross Island for their help and support.

Abstracts

TREMATODE INFECTION AFFECTS INTERTIDAL SNAIL ATTACHMENT STRENGTH

K. O’DWYER AND R. POULIN

Department of Zoology, University of Otago, Dunedin 9054, New Zealand

Attachment strength is a key determinant of survival for rocky shore gastropods, as getting dislodged by wave action or predators has negative consequences. Yet little is known of the factors that cause inter-individual variation in attachment strength among conspecifics. In this study, we tested the influence of infection by trematodes on the suction-mediated attachment strength of periwinkles from two New Zealand species, Austrolittorina cincta and A. antipodum. Using a standardized experimental protocol, we measured both the strength of attachment of individual snails to the substrate, and its repeatability, i.e. the consistency of measurements taken on different occasions on the same individuals. We then compared the attachment of snails infected with a philophthalmid trematode with that of their uninfected conspecifics. First, we found that for a given snail mass, infected snails were more easily detached from the substrate than uninfected ones, although this pattern was only significant for Austrolittorina cincta, the larger of the two snail species. Second, the repeatability of attachment strength measurements per individual snail did not differ between infected and uninfected conspecifics, for either of the two periwinkle species. Our findings show that parasitism can weaken snail attachment, and indirectly increase snail mortality, on exposed rocky shores, suggesting a new way in which parasites can affect host population dynamics.

Abstracts

CRYPTIC SPECIES DIVERSITY IN PARACARYOPHYLLAEUS (CESTODA: CARYOPHYLLIDEA), PARASITES OF LOACHES (COBITIDAE) IN EURASIA

1 2 1 2 M. OROS , T. SCHOLZ , E. BAZSALOVICSOVÁ AND J. BRABEC

1Institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 040 01 Košice, Slovakia 2Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic

Molecular phylogenetic analysis of an extensive collection of monozoic tapeworms of the genus Paracaryophyllaeus Kulakovskaya, 1961 (Cestoda: Caryophyllidea), parasites of loaches (Cypriniformes: Cobitidae) in Eurasia, has revealed cryptic species diversity within this long-time monotypic genus, especially in the P. gotoi (Motomura, 1927) species complex [syn. P. dubininorum (Kulakovskaya, 1961); type species). Three independent, well- supported clades were discovered on the basis of molecular data: (i) specimens from Misgurnus anguillicaudatus and Cobitis lutheri from China, Russian Far East and Japan – called herein Paracaryophyllaeus cf. gotoi 1, which may be conspecific with P. gotoi (Motomura, 1927), although in the absence of sequence data for P. gotoi from its type locality (basin of the River Kumkan in Korea), no certain inferences about their identity can currently be made; (ii) specimens from M. anguillicaudatus from China and Japan – Paracaryophyllaeus cf. gotoi 2, which are morphologically indistinguishable from those of Paracaryophyllaeus cf. gotoi 1; and (iii) morphologically distinct tapeworms from the endemic loach Cobitis bilseli from southwestern Turkey (Beyşehir Lake), which are described herein as a new species. Paracaryophyllaeus n. sp. differs from the remaining species of the genus in the following characteristics: the testes begin anterior to the first vitelline follicles (versus posterior), the body is short and robust (versus more elongate and slender), and the scolex is wide, rounded or apically tapered (versus claviform to truncate).

Acknowledgements: Slovak Research and Development Agency (Project No. APVV-0653-11), Grant Agency VEGA (No. 2/0129/12), Czech Science Foundation (Project No. P505/12/G112) and Institute of Parasitology (RVO: 60077344).

Abstracts

THE IMMUNITY AND PARASITISM IN GIBEL CARP (CARASSIUS GIBELIO), A SPECIES WITH DUAL FORM OF REPRODUCTION

T. PAKOSTA AND A. VETEŠNÍKOVÁ ŠIMKOVÁ

Department of Botany and Zoology, Faculty of Science, Masaryk University, Kamenice 753/5, 625 00 Brno

The gibel carp, Carassius gibelio (Cyprinidae) is a unique fish species with special reproduction strategy i.e. forming sexual lineages and gynogenetic lineages. The original invasive population of gibel carp entering into Czech Republic in the eighties of the twentieth century was composed of only triploid females with gynogenetic reproduction. Recently, gynogenetic form (mainly composed of triploid females but small proportions of triploid males and even of tetraploid males have been reported) coexists often together with the sexually reproducing diploid form in the same habitats. On the basis of different published sources, C. gibelio is recently considered as either a species or sub-species belonging to C. auratus complex. Previous studies showed that even morphologically similar specimens of C. auratus complex may belong to four different mitochondrial lineages (i.e. C. gibelio, C. auratus, C. landsdorfi and lineage M). Thus, the first aim of our study is to determine the mitochondrial lineage of morphologically determined C. gibelio. The next aim of our study is to evaluate the metazoan parasite load in diploid sexually reproducing form and triploid gynogenetic form (both of the same age) of C. gibelio (i.e. the representatives of C. gibelio mt lineage) sampled in the same locality situated in Dyje River. In addition, the selected parameters of non-specific immunity are evaluated i.e. respiratory burst and lysozym activity and compared between two reproductive forms hypothesizing that the difference in parasitism and immunity may at least partially explain the evolutionary shift from gynogenetic toward sexually reproducing forms.

Acknowledgements: Czech Science Foundation (Project No. P505/12/0375).

Abstracts

ORGANOGENESIS OF REDIA AND CERCARIA OF FASCIOLOIDES MAGNA (DIGENEA)

1 1,2 1 J. PANKRÁC , M. KAŠNÝ AND P. HORÁK

Fascioloides magna (Digenea) is two-host parasite causing severe destructions of the liver tissue of large mammals, in the Czech Republic e.g. to Cervus elaphus. The intermediate hosts of F. magna are the freshwater snails of the family Lymnaeidae, which are in Europe represented by compatible semiaquatic species Galba truncatula. The development of F. magna in the intermediate host includes the asexual reproduction of two morphological types of larval stages - sporocyst and redia leading to a final production of larva infective for the definitive host - cercaria. In order to get more information about this phase of development we focused on organogenesis of three organ systems - musculature, nervous system and excretory system of two F. magna larval stages - redia and cercaria. Daughter rediae and cercariae at various level of development were isolated from living larvae of the previous generation, fixed and labelled. Visualization was performed by using the fluorescent and confocal microscopy. Development of organ systems was followed from germinal balls up to the developed individuals of rediae and cercariae. The knowledge of organogenesis of trematodes is still limited, therefore our results will be new not only for F. magna but also for the trematodes in general.

Acknowledgements: Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013).

Abstracts

ARE THE LIZARDS FROM BRAZIL “GOOD FRIENDS” OF NEMATODE TAXONOMISTS? AN ANSWER BASED UPON THREE PERSONAL CASES.

1 2 F.B. PEREIRA AND J.L. LUQUE

1Program of Post Graduation in Veterinary Sciences, Federal Rural University of Rio de Janeiro, Seropédica, BR 465 Km 7, Rio de Janeiro, Brazil 2Department of Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, BR 465 Km 7, Rio de Janeiro, Brazil

Lizards represent a widespread group of squamate reptiles with incredible plasticity of habitat exploration and feeding habits; diversity of behaviour and morphological adaptations; frequent variability in ontogeny and sexual dimorphism. In Brazil, these organisms are found in all national land biomes and its complex ecosystems. For these reasons, lizards are ideal models for ecological studies and, consequently, for parasitological analysis. Besides it is assumed that the parasite communities found in reptilian hosts are, generally composed by low species richness, little is known about the diversity and taxonomy of these small organisms. In addition, there is a lack of experts on the taxonomy of parasites in reptiles throughout the world. Among the parasites of lizards from Brazil, the most representative and diverse group has been composed by nematodes. Because of that, and together with the lack of taxonomist experts on nematode parasitizing reptiles in Brazil, a question can be raised: Are the lizards “good friends” of nematode taxonomists? If the answer is positive, why there is no real specialist in this field of work in Brazil? If the answer is negative, we strongly think that all the evidences show the contrary. In fact, there is no simple answer for this question, because that depends upon the point of view. We present here three cases, in which we studied the parasite fauna of two different species of lizards, in order to defend our point of view. In the first case, we studied the parasite component community of the small tropidurid lizard Tropidurus torquatus (Tropisuridae) in a rocky outcrop area (Atlantic Rainforest biome) located in the State of Minas Gerais, Brazil. We found four different species of nematodes, of which two were described as new species namely, Oswaldofilaria chabaudi (Onchocercidae) and Parapharyngodon bainae (Pharyngodonidae), and also one physalopterid nematode supposed to be unknown. Lastly, we found Physaloptera lutzi (Physalopteridae) a species requiring a redescription, which is under finalization. It is worth noting that T. Torquatus is one of the most studied lizards for parasite community ecology and taxonomy in Brazil. In the second case, we studied some physalopterids collected from the large teiid lizard Salvator merianae (Teiidae) in a wild area from the same biome fragment studied in the first case. In S. merianae we found a new species belonging to the broad genus Physaloptera which was described as P. tupinambae. In the third case, we studied other physalopterids parasitic in another S. merianae, but now from a fragment of urban forest in the municipality of Juiz de Fora, State of Minas Gerais. This material revealed to be another new species of Physaloptera namely, P. bainae, which has a completely different morphology from that of P. tupinambae. Thus, in only one geographical locality and in two different species of lizards, we were able to find six taxonomical gaps on nematodes, of which four (until then) resulted in description of new species and one will result in a redescription with new important morphological data. After this quick exposition, how would you respond the proposed question? If you are a nematode taxonomist we can imagine your answer...

Abstracts

ISOFORMS OF A CATHEPSIN L-LIKE PEPTIDASE IN CERCARIAE OF DIPLOSTOMUM PSEUDOSPATHACEUM

T. PERHÁČOVÁ AND L. MIKEŠ

Department of Parasitology, Faculty of Science, Charles University in Prague, Viničná 7, 128 44 Prague 2, Czech Republic

Cercarie of the widespread fish eye fluke Diplostomum pseudospathaceum are able to invade the skin of fish, the second intermediate host, by active penetration. For this purpose they are equipped with penetration glands. The content of these glands is in charge of tissue disturption, extracellular matrix degradation etc. We have purified and characterized a major 24 kDa cysteine endopeptidase from cercarial soluble protein extracts. Test with relevant inhibitors and fluorogenic substrates confirmed that the purified protein is a cathepsin L-like peptidase. Besides their different pI, peptidases in particular FPLC fractions (cation exchange chromatography) slightly differed in pH optima of peptidolytic activity and in electrophoretic mobility. The results of mass spectrometry analysis which was applied on particular fractions suggested that there is more than one cathepsin L isoform involved. Therefore we used degenerate PCR primers based on consensual sequences of active sites of cysteine peptidases and cDNA from sporocysts with developing cercariae to amplify corresponding genes. Following cloning we obtained partial sequences of four different cathepsin L-like isoforms. These will be expressed as recombinant proteins and characterized in terms of their localization and biological functions.

Abstracts

UNIQUE DIPHYLLOBOTHRIIDEAN TAPEWORMS FROM REPTILES

I. POKORNÁ AND R. KUCHTA

Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic & Faculty of Science, University of South Bohemia, 370 05 České Budějovice, Czech Republic

The diphyllobothriid tapeworms are well known parasites of mammals including man, but less know as parasites of reptiles. These tapeworms belong to the family Scyphocephalidea and compose three genera (Bothridium, Duthiersia, Scyphocephalus) characteristic with unique scolex morphology. Two genera are specialised to varanid lizards and the genus Bothridium is parasite of snakes. All genera are known from Asia, two from Africa (Bothridium and Duthiersia) and one from Australia and South America (Bothridium). The genera are well characterised, but species composition of scyphocephalids is not well understand. This study revised available data on scyphocephalid tapeworms and newly obtained material from Ghana, Vietnam, Philippines and Thailand.

Abstracts

THE PROTEIN INHIBITORS OF PEPTIDASES FROM EUDIPLOZOON NIPPONICUM AND THEIR PREPARATION IN RECOMBINANT FORM BY USING THE CELL-FREE EXPRESSION SYSTEMS

1 1 1 1,2 P. ROUDNICKÝ , J. ILGOVÁ , M. GELNAR AND M. KAŠNÝ

The cell-free (CF) protein expression systems are useful applications in molecular biology enabling the rapid production of lower amount of selected protein molecule. They represent the alternative to “classical” expression systems adopting the whole live cells, e.g. bacteria - Escherichia coli or yeast - Pichia pastoris. The cell-free expression methods have been used since 1960 and up to date they were significantly improved; a number of commercial CF kits is now available. The proteolytic enzymes – peptidases are essential molecules not only for helminths but also for each living organisms. They are generally participating e.g. in digestive process, in modulation of host immune reaction or in evasion of host immune response. The proteolysis is at the level of the organism well controlled process regulated by number of parasite/hosts specific molecules, including inhibitors – serpins. Number of extra- or intra-cellular serpins was identified from plants, animals, viruses, and microbes. Serpins participate in fundamental physiological processes, such as blood coagulation, fibrinolysis, inflammation, signaling, immune responses and hormone carriage. This suggested their important role in host-parasite interaction. Our work is focused on the characterization of inhibitors (serpins) of serine peptidases, which were identified in Eudiplozoon nipponicum (cyprinid fish ectoparasite) transcriptome data.

Acknowledgements: Czech Science Foundation (Project Nos. P506/12/1258, GBP505/12/G112) and Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013).

Abstracts

SHOW ME YOUR SPINES – DIPLOSTOMUM SPP. DIVERSITY IN THE RUHR RIVER IN GERMANY

1 2 2 2 1 C. SELBACH , M. SOLDÁNOVÁ , S. GEORGIEVA , A. KOSTADINOVA AND B. SURES

1Department of Aquatic Ecology, University of Duisburg-Essen, Universitätsstraße 5, D-45141, Essen, Germany 2Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic

Trematodes of the genus Diplostomum are widely distributed fish parasites with a serious impact on aquaculture and wildlife management. The parasites’ complex life-cycles involve lymnaeid snails and fish as intermediate hosts and fish-eating birds as definitive hosts. Due to the high morphological similarities between species at all stages of the life-cycle, the taxonomy of Diplostomum is still in a controversial state. While the free-living cercariae are considered to be the most important life-stage for morphological species identification, morphological studies of infections in snails from natural populations are scarce. Recent molecular studies from North America and Europe have revealed much higher species diversity than previously estimated from morphology, highlighting the need for integrated approaches to close this gap. In order to assess the diversity of trematodes in natural snail populations in a reservoir system of the Ruhr River, Germany, both morphological and molecular identification approaches were applied. Snails were collected by hand in five reservoirs and examined for patent and pre-patent trematode infections between May 2012 and September 2013. A total of 3,181 snails of four lymnaeid species, Radix auricularia (1,919), Radix peregra (349), Lymnaea stagnalis (245) and Stagnicola palustris (668) were examined and revealed 78 infections (2.5 %) with Diplostomum spp. All Diplostomum spp. isolates were photographed and preserved in 96% ethanol and 4% formaldehyde solution for molecular and morphological analyses. Cox1 and ITS1-5.8S-ITS2 sequences from 52 isolates were generated and aligned with available European sequences from GenBank. Analyses of the sequences revealed a diverse Diplostomum fauna with seven genetically distinct lineages: two described species, D. pseudospathaceum (in L. stagnalis and S. palustris) and D. spathaceum (in R. auricularia), plus five species-level lineages of the "D. mergi" species complex in R. auricularia, including Clade Q of Georgieva et al. (2013). Detailed analyses of the formaldehyde-fixed isolates by SEM revealed distinctive features of cercarial morphology (e.g. body spination) that are central to species delineation but not accessible under light microscopy. The diversity within this single genus highlights the need to carefully take morphological and molecular approaches into consideration for a better understanding of the parasites’ complex taxonomy.

Abstracts

EFFECTS OF INFECTION WITH TRICHOBILHARZIA REGENTI ON HAEMOCYTE ACTIVITIES OF THE HOST SNAIL RADIX LAGOTIS

1 1,2 1,3 4 1 V. SKÁLA , Z. JINDROVÁ , M. VOSTRÝ , A.J. WALKER AND P. HORÁK

1Charles University in Prague, Faculty of Science, Department of Parasitology, Viničná 7, 128 44, Prague 2, Czech Republic 2Charles University in Prague, 1st Faculty of Medicine, Institute of Immunology and Microbiology, Studničkova 7, 128 43, Prague 2, Czech Republic 3Institute of Haematology and Blood Transfusion, U Nemocnice 2094/1, Prague 2, 128 20, Czech Republic 4Molecular Parasitology Laboratory, School of Life Sciences, Kingston University, Penrhyn road, Kingston upon Thames, KT1 2EE, United Kingdom

Compatibility between snails and trematode parasites is a fascinating phenomenon that is determined by several factors. These factors remain largely unknown for Radix lagotis- Trichobilharzia regenti combination. We focused on snail haemocytes, mobile phagocytic cells that mainly ensure cellular immune responses against the invading parasites. A combined light and electron microscopic observations revealed that haemocytes of R. lagotis are capable to surround but not destroy developing larvae of T. regenti. Consequently, the infection progressed to a patent phase when haemocyte activities such as hydrogen peroxide (H2O2) production or phagocytosis decreased in infected snails when compared to uninfected counterparts. Only the number of haemocytes increased as a result of parasite infection. At the molecular level, enzymatic signalling cascades leading to activation of protein kinase C (PKC) and extracelullar-signal regulated kinase (ERK) were found to be crucial in regulation of haemocyte mediated H2O2 production and phagocytosis, respectively. Both PKC and ERK were less active in haemocytes from infected snails than those of uninfected ones. Furthermore, homogenised T. regenti cercariae affected PKC and ERK phosphorylation (activation) in haemocytes from uninfected snails. The observed reduction of H2O2 production and phagocytosis by haemocytes from infected snails might, therefore, be caused (at least partly) by lowered PKC and ERK activity in these cells that likely facilitates survival of T. regenti in R. lagotis.

Acknowledgements: Charles University in Prague (Project No. 435911, research programmes PRVOUK - P41/PrF, UNCE - 204017 and SVV 260074/2014).

Abstracts

ANTICOAGULATION FACTORS AND BLOOD UPTAKE BY MONOGENEANS OF THE FAMILY DIPLOZOIDAE

K. SKIPALOVÁ AND L. MIKEŠ

Department of Parasitology, Faculty of Science, Charles University in Prague. Viničná 7, 128 44 Prague 2, Czech Republic

Monogeneans of the family Diplozoidae are blood-feeding ectoparasites occupying the gills of cyprinid fishes. In general, hematophagous parasites must overcome host's hemostasis (blood coagulation) to facilitate blood intake using anticoagulant peptides and anti-platelet aggregation agents. Although antihemostatic molecules have not been reported yet from monogeneans, their presence is highly probable in sanguinivorous species. These are probably produced by specialized glands and will inhibit thrombosis and clot formation. Antihemostatic molecules often act as protease inhibitors that target one or more serine proteases comprising the coagulation cascade. This study is focused on biochemical and molecular characterization of potentional anticoagulants involved in blocking of factors in fish coagulation cascade. We measured inhibitory activity of excretory-secretory products and homogenates (Eudiploozon nipponicum, Paradiplozoon homoion and Paradiplozoon bliccae) towards coagulation factors IIa and Xa with negative results. Employing transcriptomic data of E. nipponicum and bioinformatics we found three protein groups of potential anticoagulants - annexins, serpins and Kunitz-domain proteins. For further analyses we focused on the Kunitz protein family. In the transcriptomic data, all Kunitz sequences were identified to be only partial. These Kunitz proteins contain one or more structurally related active domains, which are able to inhibit the function of proteases. In addition, we described similarities between Kunitz sequences from Eudiploozon nipponicum and their homologues from other hematophagous parasites, which exhibit anticoagulant activity (e.g. Ixodes scapularis, Rhipicephalus microplus or Haemaphysalis longicornis). To obtain the full length Kunitz sequences, we used Rapid Amplification of cDNA Ends (3´ and 5´ RACE PCR). Separate Kunitz domains will be prepared in the recombinant form for further tests.

Acknowledgements: Czech Science Foundation (Project No. P506/12/1258).

Abstracts

IMPORTED SYSTEMIC HELMINTHIC INFECTIONS

F. STEJSKAL

1st Department of Infectious Diseases, 2nd Faculty of Medicine, Charles University and Hospital Na Bulovce, Prague, Czech Republic Department of Infectious Diseases, Regional Hospital Liberec, Czech Republic Institute of Immunology and Microbiology, 1st Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic

Systemic (tissue) helminthic infections represent common human infections in tropics. Large proportion of world population is living in the risk of schistosomiasis, filariasis, echinococcosis and cysticerciasis. These infections are characterized by migration of adult worms or their larval stages through the internal organs of the host. Tissue helminthoses are chronic diseases with low mortality but substantial morbidity and long-term sequel. The acute stage of infection may be connected with the systemic immune reaction based on immune complex production and it is presented as fever, rigor, headache, muscle and joint pain, the respiratory symptoms (cough, dyspnoe, chest pain) and high eosinophilia. During last ten years there were investigated at our departments around 16 patients with schistosomiasis, 10 patients with cystic echinococcosis (CE), 4 patients with alveolar echinococcosis (AE), 3 patients with strongyloidiasis and several patients with suspected trichinellosis. Systemic tropical filarial and fluke infections or cysticercosis are rarely imported to the Czech Republic (CZ). Schistosomiasis is the most common severe systemic helminthosis imported to CZ and all our patients visited or lived in sub-Saharan Africa. Six patients presented with systemic febrile illness (Katayama fever). Travel history, contact with fresh water and eosinophilia suggested schistosomiasis which was confirmed by serology, parasitological investigation of stool and urine was negative in these patients. Four cases of urinary schistosomiasis were diagnosed by parasitological urine investigation or by cystoscopy and histology. Five patients had chronic intestinal schistosomiasis, three with parasitologically confirmed infection and two with positive serology only. Eosinophilia was presented in the large majority of our patients with schistosomiasis. Eight of ten our CE cases were diagnosed in migrants from endemic regions: Bulgaria (2x), Romania, Russia, Kazakhstan, Montenegro, Tajikistan, and Uzbekistan. Two CE patients are Czechs. A 36-year old man had a history of short-term visits of Bulgaria, Russia and Mongolia since late 1990s and a 64-year female acquired infection in CZ, probably. She lived in the rural region in Eastern Bohemia and did not travel to the CE endemic countries. The CE cysts have been identified in liver in 8 patients, in lungs only in 1 case, and in both liver and lung in 1 patient. Currently, four female patients with advanced inoperable liver AE are in long-term follow-up at our departments. Due to the disease progression and local complications one patient has undergone the orthotopic liver transplantation in 2013. All our AE patients are on the long-term continuous therapy with albendazole that is well tolerated. Significant eosinophilia was presented in two patients with CE only and two of four AE patients at the time of diagnosis. The elevation of total IgE immunoglobulin was recorded in five patients with CE and two patients with AE. Specific antibodies were elevated or at least borderline in all our patients with CE and AE. There is one patient in the long-term follow up at our department with prolong hypereosinophilia and probable lymphatic filariasis after visiting West Irian in 2007. Imported onchocerciasis was diagnosed in traveler to Cameron and cysticercosis in 6-year old boy living with parents in India for 4 years.

Abstracts

THE EXPRESSION PROFILE OF CATHEPSIN L IN DEVELOPMENTAL STAGES OF FASCIOLOIDES MAGNA

1 1 1 1,2 R. ŠAŠKOVÁ , R. LEONTOVYČ , P. HORÁK AND M. KAŠNÝ

Fascioloides magna is an important liver fluke which has originally been parasitising the North American ruminants. In 19th century it was introduced to Europe and now is common in several European countries including Czech Republic, where infects predominantly Cervids (locally prevalence reaches 95%) and occasionally Bovids. During our research on F. magna we investigated the composition of excretory-secretory products (ESP) of adult worms and we identified some molecules which are probably essential for life of this parasite, e.g. cathepsin L. Cathepsin L is the most abundant proteolytic enzyme in the ESP, it belongs to a cysteine-peptidase family and we suppose its participation in numerous biological processes such as e.g. digestion of blood, host tissue invasion and evasion of host immune response. Since the cysteine-peptidases are in general multifunctional enzymes their expression level can vary according to parasite requirements during a life cycle. By using the molecular methods (PCR), cathepsin L was identified in all F. magna life stages. In order to explore the expression profile of cathepsin L during the parasite’s life cycle we adopted qPCR methods. All necessary preliminary steps and qPCR assays were strictly designed according to MIQE guidelines to keep a high standard of reproducibility of data and to prevent the misinterpretation of the results.

Acknowledgements: Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013, GAUK 502313).

Abstracts

CYSTATINS OF THE PARASITIC NEMATODES FROM THE GENUS TRICHINELLA

1 1,2 1 3 1 1,2 L. ŠKORPÍKOVÁ , E. ŠRÁMOVÁ , J. ILGOVÁ , B. KOUDELA , M. GELNAR AND M. KAŠNÝ

1Department of Botany and Zoology, Faculty of Science, Masaryk University Brno, Kamenice 753/5 625 00 Brno Bohunice, Czech Republic 2Department of Parasitology, Faculty of Science, Charles University in Prague, Vinicna 7, 128 00 Prague, Czech Republic 3Department of Pathological Morphology and Parasitology, Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Science Brno, Palackého 1/3, 612 42 Brno, Czech Republic

The nematodes of the genus Trichinella are intracellular parasites of small intestine and skeletal muscle cells. They can infect a broad range of worldwide distributed hosts like birds, reptiles and mammals. All Trichinella spp. are zoonotic but only six of twelve genotypes were detected in humans, causing the trichinellosis - a serious, sometimes fatal, human disease, which has been documented in 55 countries of the World. Our study is focused on characterization of composition of excretory-secretory products (ESP) from L1 muscle larvae of three Trichinella species - Trichinella spiralis, T. britovi and T. pseudospiralis. It was recorded that among the dominant components of ESP from L1 larvae belong the cystatins. These proteins are reversible inhibitors of proteolytic enzymes - cystein peptideases and they are involved in many relevant biological processes such as development, growth, digestion, migration of parasites and suppression of the host immune responses. The main aim of our study is to prepare the recombinant form of three trichinella cystatins and characterize/compare their properties. We designed primers specific for “multi cystatin-like domain” gene of T. spiralis. Subsequently, we obtained sequences of this gene for the other two species and we compared these data. We found out that cystatin genes of T. spiralis and T. britovi have high sequence homology although gene of T. pseudospiralis is more different (e.g. in number of domains or conserved motifs).

Acknowledgements: Ministry of Education, Youth and Sport Czech Republic (Project CONTACT II No. LH12096) and Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013).

Abstracts

DANCING CHROMOSOMES: KARYOTYPE VARIATION IN PARADIPLOZOON HOMOION (MONOGENEA, DIPLOZOIDAE)

1 1 2 M. ŠPAKULOVÁ , M. BOMBAROVÁ AND B. KOUBKOVÁ

1Institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 040 01 Košice, Slovakia 2Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic

Individuals of Paradiplozoon homoion, the unique gill parasite of variety of cyprinid fishes, were investigated karyologically using classical techniques and fluorescent in situ hybridisation approach. Dividing gametogonial cells of individual parasites showed a conspicuous polymorphism associated with the fission of one or both homologue metacentric chromosomes of the first pair that occur in different frequencies in individual worms and various fish hosts from geographically distant regions. The rest of the chromosome complement was steadily composed from 5 shorter pairs with predominating subterminal centromeres. As a result, the karyotype revealed three cytotypes. First, 2n = 12 with one large metacentric and 5 subtelocentric pairs, second the ancestral type 2n = 14 with 7 subtelocentric pairs, and third, intermediate type with 2n = 13 with one large metacentric chromosome plus a pair of non-homologous subtelocentrics plus 5 subtelocentric pairs). Various cytotypes occurred even at one locality, one fish host and even one worm individual, although 2n = 12 cytotype predominated in Gobio gobio and the 2n = 14 in Barbus barbus. Parasites of Rutilus rutilus differed in cytotype frequency depending on locality. The extraordinary variability was found also in number of loci for ribosomal genes: one rDNA locus was regularly located in the site of secondary constriction of the penulimate chromosome pair; the second one occurred sometimes at its second larger neighbour. The course of meiosis was not substantively different in the three cytotypes except for the shape of the bivalents in metaphase I. Large metacentric pair of the first cytotype formed round bivalent with two chiasma while all other chromosomes formed end-to-end bivalents. Taking into account the present and previous karyological data on P. homoion, it can be hypothesized that the parasite populations reveal unbalanced chromosomal evolution, however, the partial but apparent linkage of the karyologically distinct lineages to different fish hosts (gudgeon versus other cyprinids) revives the existing hypothesis on P. homoion as a complex of species.

Acknowledgements: Slovak projects VEGA (Project Nos. 2/0168/13, APVV-0653-11) Czech Science Foundation (Project No. GAP506/12/1258).

Abstracts

AFLP: STUDY OF GENETIC VARIABILITY IN TRICHINELLA POPULATIONS

1,2 1,3 1 1,2 E. ŠRÁMOVÁ , D. JIRSOVÁ , M. GELNAR AND M. KAŠNÝ

1Department of Botany and Zoology, Faculty of Science, Masaryk University Brno, Kamenice 753/, 625 00 Brno Bohunice, Czech Republic 2Department of Parasitology, Faculty of Science, Charles University in Prague, Vinicna 7, 128 00 Prague, Czech Republic 3Institute of Parasitology, Biology Centre, ASCR, v.v.i., Branišovská 31, 37005 České Budějovice, Czech Republic

Parasitic worms from the genus Trichinella are well known for their worldwide veterinary and medical importance. These organisms are able to parasitize in both carnivorous and omnivorous vertebrates, including man. The infection initially develops after the ingestion of raw or undercooked infected meat. Contrary to human infections, which may lead to death, there are usually no recognized clinical signs within trichinella positive animals. Twelve Trichinella taxa are divided into two clades characterized by the presence or absence of an intramuscular collagen capsule. In the Czech Republic we recognize three trichinella species - Trichinella spiralis, T. britovi (mainly in carnivores and wild boars) and T. pseudospiralis (mainly in predatory birds) are found in wild animals. While T. spiralis and T. britovi induce the formation of a collagen capsule surrounding the parasite, T. pseudospiralis survives in a non-encapsulated form. The main aim of our work is to compare the genetic diversity between three Trichinella European species. AFLP - Amplified Fragment Length Polymorphism is a method, which can reveal the PCR fragments´ length variability by using the DNA fingerprinting. By the sequencing of individual amplicons, the taxonomic membership and genetic plasticity of the populations of trichinella species can be examined and results can also documented the geographical origin of trichinella populations. We applied classical molecular phylogenic methodology based on cytochrome oxidase I (coxI) and NADH dehydrogenase subunit 3 (nad3). The nucleotide sequences of T. spiralis and T. britovi nad3 and coxI genes were recorded and showed high uniformity (variance was low). The AFLP approach is in process and up to now we obtained the first data from fragmentation analysis.

Abstracts

ANTIGENS OF BIRD SCHISTOSOMES OF THE GENUS TRICHOBILHARZIA: PREPARATION OF RECOMBINANT PROTEINS AND DUCK ANTIBODY REACTIVITY

1 1,2 1 L.TURJANICOVÁ , M. KAŠNÝ AND L. MIKEŠ

1Department of Parasitology, Faculty of Science, Charles University in Prague, Viničná 7. 12844 Prague 2, Czech Republic 2Institute of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic

Bird schistosomes of the genus Trichobilharzia are dixenous trematodes bound to aquatic environments. Although waterfowl serve as specific hosts, schistosome invasive larvae (cercariae) can also penetrate into mammalian hosts including humans where they may cause an itchy hypersensitive skin response, cercarial dermatitis. Currently, there is no specific non- invasive method for diagnosis of infections caused by bird schistosomes. Previous studies dealing with the immune response of nonspecific murine hosts revealed two antigens strongly recognized by IgG1 and IgE antibodies, characterized as trioso phosphate isomerase (MW 25 kDa) and glyceraldehyde 3-P dehydrogenase (MW 34 kDa). The focus of our recent research was the antibody responsivenes of ducks to selected antigenic structures, such as those described above, in order to prepare diagnostic markers suitable for diagnosis of avian trichobilharziases or cercarial dermatitis. We designed degenerated primers based on partial amino acid sequences of trioso phosphate isomerase of Trichobilharzia szidati and performed PCR. The obtained nucleotide sequence was in 82% similar to a TPI sequence of Schistososma heamatobium. Subsequently, the recombinant protein was produced by using Escherichia coli bacterial expression system and tested in ELISA and Western blots with sera of ducks experimentally infected by trichobilharziae. Futhermore, we checked the reaction of antibodies from infected specific hosts against recombinant cathepsin B1.1 and cathepsin B2 from Trichobilharzia regenti. The first experiments with these recombinant antigens showed promissing results.

Acknowledgements: Czech Science Foundation (Project No.13-29577S) and Charles University in Prague (Project No. GAUK 243-259305 B-Bio/2014).

Abstracts

CHARACTERIZATION OF SCHISTOSOMA MANSONI SERINE PROTEASE 2 (SMSP2)

1,2 1,3 4 1 L. ULRYCHOVÁ , A. LEONTOVYČ , M. HORN AND J. DVOŘÁK

1Institute of Molecular Genetics of the ASCR, Videnska 108, Prague 14220, Czech Republic 2Department of Parasitology, Faculty of Science, Charles University, Vinicna 7, Prague 12844, Czech Republic 3Department of Biochemistry, Faculty of Science, Charles University, Hlavova 8, Prague 12843, Czech Republic 4Institute of Organic Chemistry and Biochemistry of the ASCR, Flemingovo namesti 2, Prague 16610, Czech Republic

Proteolytic activities are integral part of many physiological processes and in schistosomes are also crucial in host-parasite interaction in processes such as successful invasion, survival and reproduction of parasite. Besides of relatively well characterized enzymes of Schistosoma mansoni there are groups of proteases which are being neglected. Our research is focused on serine proteases from trypsin-like family S1 from clan PA. S1 proteases are the major cornerstone of the vertebrate degradome. In human they are responsible for highly important vital processes such as blood coagulation, fybrinolysis, immunity and apoptosis. Interestingly, some schistosomal S1 peptidases share significant similarities with human host ones although schistosomes carry six times less genes with features of S1 family than human. This together with fact that schistosomes actively interact with host environment via their own proteolytic apparatus led us to the hypothesis that serine proteases might play essential role in immuno-modulatory strategies of the parasite. We were able to partially described trypsin-like S. mansoni serine protease 2 (SmSP2). According to our data SmSP2 is highly expressed in life-stages residing in definitive host (schistosomula, adult worms). We employed RNA in situ hybridization and polyclonal antibodies in order to localize the gene expression and enzyme localization. In addition, several modifications of recombinant protein were expressed in Escherichia coli and Pichia pastoris expression systems. Purified recombinants were submitted to several refolding techniques to obtain active proteases when necessary. Active recombinants were characterized using synthetic peptidyl-substrates. Finally, the substrate activity screening assay was utilized to identify enzymatic substrate specificities and optimal substrates. First results will be presented and the implications of these findings will be discussed.

Acknowledgements: Ministry of Education, Youth and Sports of the Czech Republic (Project CONTACT II No. LH12096) and Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013).

Abstracts

MOLECULAR AND SEROLOGICAL DIAGNOSIS OF INFECTIONS CAUSED BY TRICHOBILHARZIA

M. VAŠČIKOVÁ AND P. HORÁK

Department of Parasitology, Faculty of Science, Charles University in Prague Viničná 7, 128 44 Prague 2, Czech Republic

Cercariae of the genus Trichobilharzia are able to penetrate the skin of accidental hosts and cause an acute inflammatory response known as cercarial dermatitis. Not all schistosomula are necessarily trapped and destroyed. They are able to escape from the host skin and migrate to the internal organs. Fortunately, they are not able to complete their development in the accidental host and die a few days after infection. The goal of our experiments is to detect parasite DNA (T. regenti and T. szidati ) in serum and cerebrospinal fluid of ducks and mice c57BL/6 using commercial kits and PCR. For this method we designed specific primers for different genes (ITS 1, cox1 and others). We test the same biological material, homogenate of cercariae from T. regenti and T. szidati, as antigen using ELISA and Western blot. Changes in the levels of IgG and IgM are measured at different days after infection by ELISA (5-100 days, depending on the animal). Using Western blot, we try to identify antigens recognized by IgG and IgM from sera of infected mice and humans. A specifically recognized antigen was identified in the study by Lichtenbergová et al. (2008). The study dealt with the antibody response against antigens of T. regenti in experimentally infected mice c57BL/6 and humans with history of cercarial dermatitis. Analysis of sera from both hosts revealed that IgG and IgE antibodies recognized 34kDa antigen that was identified in the homogenate of cercariae and cercarial excretory/secretory products. Therefore, we repeated the test and the 34-35kDa antigen was also recognized by IgG antibodies. According to Lichtenbergová et al. (2008), this antigen could serve as a possible diagnostic marker. Our intention is to separate this antigen by 2D electrophoresis and then identify a protein spot representing the antigen by mass spectrometry.

Abstracts

NEMATODES OF THE GENUS CUCULLANUS MÜLLER, 1777 IN THE NEOTROPICS: PROBLEMS AND PERSPECTIVES

1 2 1 F.M. VIEIRA , T. SCHOLZ AND J.L. LUQUE

1Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, Seropédica, BR 465 Km 7, Rio de Janeiro, Brazil 2Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic

The genus Cucullanus Müller, 1777 (Nematoda: Seuratoidea) contains more than 100 nominal species parasitizing freshwater, brackish water and marine fish worldwide; a few species were also described from marine turtles. In the Neotropical Region, 33 nominal species of Cucullanus have been reported from fish (16 species from freshwater, 14 species from marine and one species from brackish water fish). Most (19) of the Neotropical species of Cucullanus have been described from Brazil and all but two species were found only in this zoogeographical region and thus may be endemic to the Neotropics. The first Neotropical species, C. pulcherrimus Barreto, 1918, was described from Caranx lugubris Poey, 1860 (Perciformes: Carangidae) from the Trindade Island off the Brazilian Atlantic coast, but a great part (58%) of species were described in the last 20 years. This increase in the number of new taxa may correspond to the increased number of parasitological studies in the Neotropical Region, but also documents how the fauna of fish nematodes is still poorly known, not speaking about negligible knowledge of their life cycles and phylogenetic relationships. Another serious obstacle is the fact that many species of Cucullanus in the Neotropical hosts (at least 30% of nominal taxa) were inadequately described and type or voucher specimens do not exist or are in poor state. It is obvious that intraspecific morphological and genetic variability as well as the actual range of host specificity and geographical distribution of all species have to be critically assessed, preferably on the basis on evaluation of new, properly processed materials. As the first step to change this unsatisfactory situation, extensive materials of cucullanids were collected from a wide range of freshwater and marine hosts in different regions of South America and their evaluation using morphological and genetic tools is underway. Research stay of the first author in the Czech Republic is supported by a Postdoctoral fellowship from FAPERJ/CAPES (Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brazil), whereas repeated stays of T.S. in Brazil “CAPES/BRASIL Grantee” at the Universidade Federal Rural de Rio de Janeiro, Seropédica (laboratory of José Luque) were supported by the Ministério da Educaçăo (MEC), Coordenaçăo de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

Abstracts

THE BIOINFORMATIC ANALYSES OF "-OMES" OF HELMINTHS WITH SPECIAL FOCUS ON SERINE PEPTIDASES OF EUDIPLOZOON NIPPONICUM

1 2 2 2 1 1,2 J. VOREL , H. DVOŘÁKOVÁ , L. JEDLIČKOVÁ , L. MIKEŠ , M. GELNAR AND M. KAŠNÝ

Parasitic helminths are causative agents of number of serious diseases (helminthiasis) of animals and people. Our understanding to the biological processes of helminths is based also on detailed knowledge of their molecular biology and biochemistry. By using the modern pyro-sequencing methods and on-line databases (genomic, transcriptomic and proteomic) we can now faster and more accurately identify the numerous sets of protein molecules, which are essential for life of many organisms, including also parasites. Since September 2011 the “50 Helminth Genomes Project” has been running, but from a large group of Monogenea only one species is currently sequenced in detail - Protopolystoma xenopodis. We would like to change this situation and therefore we adopted Eudiplozoon nipponicum (Monogenea: Diplozoinae) as a model organism and started some preliminary analyses leading to generation of E. nipponicum genome, transcriptome and proteome data. We would like to use these data platforms for identification and further characterization of protein molecules involved into the interaction between host and parasite. Our preliminary transcriprome results suggest that such molecules could be also serine peptidases. They are essential for life of all organisms including helminthes, where they can participate e.g. on penetration, digestion and hosts immune attack evasion.

Acknowledgements: Czech Science Foundation (Project Nos. P506/12/1258, GBP505/12/G112) and Charles University in Prague (Project Nos. UNCE 204017, PRVOUK P41, SVV 267210/2013, GAUK 502313).

Abstracts

MOLECULAR AND MORPHOLOGICAL CHARACTERISATION OF CRYPTIC SPECIES OF LARVAL PLAGIORCHIIDEAN TREMATODES

1 1,2 2 2 J. ZIKMUNDOVÁ , S. GEORGIEVA , A. FALTÝNKOVÁ , M. SOLDÁNOVÁ AND A. 2 KOSTADINOVA

1University of South Bohemia in České Budějovice, Faculty of Science, Branišovská 31, 370 05 České Budějovice, Czech Republic 2Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic

Larval stages of Plagiorchis spp. are both ubiquitous and ecologically important parasites in snail populations of freshwater ecosystems in Europe. However, considerable difficulties exist in distinguishing the morphologically similar cercariae used for species identification of infections in snails. This may lead to underestimation of species diversity of Plagiorchis in these hosts. In the present study, 38 isolates of Plagiorchis spp. infecting two lymnaeid snails, Lymnaea stagnalis (L.) and Radix auricularia (L.), in five central European freshwater ecosystems were subjected to morphological and molecular assessment. Five morphologically homogeneous and genetically distinct lineages of Plagiorchis spp., namely P. elegans (Rudolphi, 1802), P. koreanus Ogata, 1938, P. maculosus (Rudolphi, 1802), P. neomidis Brendow, 1970 and Plagiorchis sp. were identified via matching molecular data for the mitochondrial cytochrome c oxidase subunit I (cox1) gene with detailed morphological and morphometric data of the cercariae. Comparative sequence analysis using partial 28S rDNA and ITS1-5.8S-ITS2 sequences revealed that three distinct cox1 lineages are conspecific with P. elegans, P. maculosus and P. koreanus, respectively, whereas the lineage identified based on cercarial morphology as P. neomidis plus a single isolate that could not be assigned to a described species, did not match any of the available sequences for Plagiorchis spp. The best morphological characters discriminating the cercariae stuidied are summarised in a key for the identification of the cercariae of Plagiorchis spp. parasitising lymnaeid populations in central Europe.

Addresses of Participants

PARTICIPANT CONTACT INFORMATION

1. Alves Vieira Philippe, Programa de Pós-Graduação em Biologia Animal, Universidade Federal Rural do Rio de Janeiro, BR 465, Km 7, Seropédica, Rio de Janeiro, 23890-000, Brazil, [email protected] 2. Barčák Daniel, Parazitologický ústav SAV, Hlinkova 3, 040 01, Košice, Slovakia, [email protected] 3. Brabec Jan, Parazitologický ústav, Biologické centrum AVČR, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 4. Bulantová Jana, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 5. Civáňová Kristina, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic [email protected] 6. Cortés Carbonell Alba, Avda. Vicent Andrés Estellés s/n, 46100 Burjassot, Valencia, Spain, [email protected] 7. Dvořáková Hana, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 8. Fajfrlík Karel, Ústav mikrobiologie LF UK a FN v Plzni, dr.E.Beneše 13, 305 00, Plzeň, [email protected] 9. Francová Kateřina, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 10. Gettová Lenka, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 11. Georgieva Simona, Parazitologický ústav, Biologické centrum AVČR, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 12. Horák Petr, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 13. Chalachanová Katarína, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 14. Chmelíková Michaela, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 15. Ilgová Jana, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 16. Jedličková Lucie, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 17. Kašný Martin, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 18. Kičinjaová Maria Lujza, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 19. Krasnovyd Vadym, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 20. Krčmářová Veronika, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 21. Kuchta Roman, Parazitologický ústav, Biologické centrum AVČR, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected]

51 Addresses of Participants

22. Leontovyč Roman, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 23. Lichtenbergová Lucie, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 24. Macháček Tomáš, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 25. Melounová Klára, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 26. Mendlová Monika, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 27. Mendoza-Palmero Carlos, Parazitologický ústav, Biologické centrum AVČR, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 28. Michálková Veronika, Ústav botaniky a zoologie AV ČR, v.v.i., Květná 8, 603 65, a PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, [email protected] 29. Mikeš Libor, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 30. O’Dwyer Katie, University of Otago, Dunedin, New Zealand, [email protected] 31. Oros Mikuláš, Parazitologický ústav SAV, Hlinkova 3, 040 01, Košice, Slovakia, [email protected] 32. Pakosta Tomáš, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 33. Pankrác Jan, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 34. Pereira Bisaggio Felipe, Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, Caixa Postal 74.540, CEP 23851–970, Seropédica, Rio de Janeiro, Brazil, [email protected] 35. Perháčová Terézia, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 36. Pokorná Ivana, Parazitologický ústav, Biologické centrum AVČR, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 37. Roudnický Pavel, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 38. Selbach Christian, Faculty of Biology, Aquatic Ecology, Universitätstr. 5, 45141 Essen, Germany, [email protected] 39. Scholz Tomáš, Parazitologický ústav, Biologické centrum AVČR, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 40. Skála Vladimír, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 41. Skipalová Karolína, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 42. Soldánová Miroslava, Parazitologický ústav, Biologické centrum AVČR, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 43. Stejskal František, I. infekční klinika, 2. LF UK a Nemocnice Na Bulovce Praha. Infekční oddělení KN Liberec, Ústav imunologie a mikrobiologie 1. LF UK a VFN Praha, Budínova 2, 180 81, Praha 8, [email protected] 52

Addresses of Participants

44. Šašková Romana, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 45. Škorpíková Lucie, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 46. Špakulová Marta, Parazitologický ústav SAV, Hlinkova 3, 040 01, Košice, Slovakia, [email protected] 47. Šrámová Eliška, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 48. Turjanicová Libuše, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 49. Uhrová Lucie, Přírodovědecká fakulta, Jihočeská univerzita v Českých Budějovicích, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 50. Ulrychová Lenka, Ústav Molekulární Genetiky AV ČR, Vídeňská 108, 142 20, Praha a Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 51. Vaščiková Michaela, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 52. Vieira Fabiano Matos, Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, Caixa Postal 74.540, CEP 23851-970, Seropédica, Rio de Janeiro, Brazil, [email protected] 53. Vetešníková Šimková Andrea, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, Czech Republic, [email protected] 54. Vojtová Terezie, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 55. Vorel Jiří, Ústav botaniky a zoologie, PřF MU, Univerzitní kampus Bohunice, Kamenice 5, 625 00, Brno, [email protected] 56. Vrbová Kristýna, Katedra parazitologie, PřF UK, Viničná 7, 128 44, Praha 2, Czech Republic, [email protected] 57. Vyhlídalová Tereza, Přírodovědecká fakulta, Jihočeská univerzita v Českých Budějovicích, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected] 58. Zikmundová Jana, Přírodovědecká fakulta, Jihočeská univerzita v Českých Budějovicích, Branišovská 31, 370 05, České Budějovice, Czech Republic, [email protected]

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21st Helminthological Days 2014. Programme & Abstracts Jan Brabec, Miroslava Soldánová & Iveta Hodová (Eds.)

Published by Masaryk University, Brno in 2014 with financial support from Academy of Sciences of the Czech Republic Graphic design: J. Brabec, M. Soldánová & I. Hodová 1st edition, 2014 Number of copies: 70 Printed by Tribun EU s.r.o., Cejl 892/32, 602 00 Brno ISBN 978-80-210-6796-7

The abstracts are published as received from authors, without any changes or corrections. The authors are fully responsible for the content and stylistic and language representation of their abstracts.