From Freshwater Fishes in Africa (Tomáš Scholz)

Home , Abbottina rivularis, Arhythmacanthidae, Blackspotted puffer, Ceratonova shasta, Cyprichromis, Diplostomum

Organizer: Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic Workshop venue: Instutute of Vertebrate Biology, Academy of Sciences CR Workshop date: 28 November 2018 Cover photo: Research on fish parasites throughout Africa: Fish collection in, Lake Turkana, Kenya; Fish examination in the Sudan; Teaching course on fish parasitology at the University of Khartoum, Sudan; Field laboratory in the Sudan Authors of cover photo: R. Blažek, A. de Chambrier and R. Kuchta

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ISBN 978-80-210-9079-8 ISBN 978-80-210-9083-5 (online: pdf)

Contents (We present only the first author in contents)

ECIP Scientific Board ...... 5 List of attendants ...... 6 Programme ...... 7 Abstracts ...... 8 Research outcomes of the Laboratory of Fish Protistology within the framework of ECIP (2012– 2017) (Astrid S. Holzer) ...... 9 Overview of the work of the Institute of Vertebrate Biology of the Czech Academy of Sciences in the ECIP project (Pavel Jurajda) ...... 11 Diversity of African monogeneans: Research outlines (2012−2018) (Eva Řehulková) ...... 13 Diversity and phylogenetic relationships of endoparasitic helminths: a brief overview of main achievements (2012–2018) (Tomáš Scholz) ...... 15 Evolutionary ecology of host-parasite interactions (Andrea Vetešníková Šimková) ...... 18 Early emerging Apicomplexa from marine invertebrates (Andrea Bardůnek Valigurová) ...... 20 From nothing to something…, “omics” of monogeneans (Jiří Vorel)...... 23 A new genus and two new species of dactylogyrid monogeneans from gills of Neotropical catfishes (Siluriformes: Doradidae and Loricaridae) (Aline A. Acosta) ...... 26 Species diversity and phylogeny of Dactylogyrus (Monogenea) parasites in Iberian Peninsula (Michal Benovics) ...... 27 RNA-seq data reveal genetic variation in virulence genes between host-associated genotypes of the parasitic cnidarian Ceratonova shasta in salmonids (Gema Alama-Bermejo) ...... 28 Towards a robust systematic baseline of Neotropical fish tapeworms (Cestoda: Proteocephalidae): amended diagnoses of two genera from the redtail catfish, Phractocephalus hemioliopterus (Phillipe V. Alves) ...... 29 Experimental evaluation of behavioural changes in gilt-head seabream infected with brain- encysted metacercariae of Cardiocephaloides longicollis (Trematoda, Strigeidae) (Gabrielle S. van Beest) ...... 30 Use of in vivo fluorescent dyes for tracking the penetration of Cardiocephaloides longicollis (Digenea: Strigeidae) into the gilthead sea bream Sparus aurata (Gabrielle S. van Beest) ...... 31 From taxonomic deflation to new cryptic species: Hidden diversity in a widespread African squeaker catfish (Dagmar Jirsová) ...... 32 Spring viremia of cyprinid species and their hybrids (Kristína Civáňová) ...... 33 Diplozoid species of endemic cyprinids from Mediterranean area (Kateřina Čermáková)...... 34 Comparative phylogenetic analysis of Apicomplexa based on 18s, 28s and contig 18s+28s rDNA (Andrei Diakin) ...... 35 Crystalline inclusions in “small” amphizoic amoebae (Iva Dyková) ...... 36 Does classification of Schilbetrema and Schilbetrematoides (Monogenea: Dactylogyridae) reflect their molecular phylogeny? (Kateřina Francová) ...... 38 Microsatellite markers – valuable tools for population genetic study of Dactylogyrus vistulae (Lenka Gettová) ...... 39

Philometrid nematodes of marine fishes from the Gulf of Hammamet, Tunisia (David G. Solís)40 The joint evolution of the Myxozoa and their alternate hosts: a history of host acquisitions and massive diversification events (Astrid S. Holzer) ...... 42 Recombinant cystatin from Eudiplozoon nipponicum (Monogenea) modulates the cytokine production by macrophages in vitro (Jana Ilgová) ...... 43 Round goby impact on native fish assemblages: a meta-analysis approach (Michal Janáč) ...... 44 Structural and molecular diversity of dactylogyrids parasitizing African characiform fishes (Maria L. Červenka Kičinja) ...... 45 Monogenean parasites of pelagic fish species in Lake Tanganyika: potential tags for host history and population structure (Nikol Kmentová) ...... 46 Fight with a jellyfish: host-parasite interaction of common parp and the myxozoan Sphaerospora molnari (Lukáš Korytář) ...... 47 Cytoskeleton and motility in the archigregarine Selenidium pygospionis (Apicomplexa) (Magdaléna Kováčiková) ...... 48 The consequences of hybridization on metazoan parasite infection level in cyprinids (Vadym Krasnovyd) ...... 49 Asian fish tapeworm: the most successful invasive parasite in freshwaters (Roman Kuchta) .... 50 Not as reduced as we thought: identification of nematocyst protein NOWA in polar capsules of Myxozoa (Jiří Kyslík) ...... 51 Myxozoa wherever you look: Uncovering myxozoan species diversity (Martina Lisnerová) ...... 53 Rasheedia n. nom. (Nematoda: Physalopteridae) for Bulbocephalus Rasheed, 1966 (a homonym of Bulbocephalus Watson, 1916), with description of Rasheedia heptacanthi n. sp. and R. novaecaledoniensis n. sp. from perciform fishes off New Caledonia (František Moravec) ...... 54 Effects of the infection of eye fluke Diplostomum pseudospathaceum on reproductive traits and metabolism of European bitterling Rhodeus amarus (Veronika Nezhybová) ...... 55 Parasite acquisition by non-native centrarchid fish Lepomis gibbosus in Europe (Markéta Ondračková) ...... 56 A new classification of Glaridacris Cooper, 1920 (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in North America, including erection of Pseudoglaridacris n. gen (Mikuláš Oros)...... 57 Factors contributing to the coexistence of two reproductive forms of Carassius gibelio in the Czech Republic. (Tomáš Pakosta) ...... 58 New genera and species of paramphistomes (Digenea, Paramphistomoidea, Cladorchiidae) parasitic in fishes from the Amazon basin in Peru (Camila Pantoja) ...... 59 Is the presence of pharmaceuticals in fish tissues associated with the abundance of parasites? (Markéta Pravdová) ...... 60 Phenotypic plasticity in Cichlidogyrus spp. (Monogenea: Dactylogyridae) parasitizing Lake Tanganyika cichlid tribes using a geometric morphometric approach: the roles of host phylogeny and locality (Chahrazed Rahmouni) ...... 61 Concentrations of fourteen trace metals in scales of three nototheniid fishes from Antarctica (James Ross Island, Antarctic Peninsula) (Kevin Roche) ...... 62 Laser microdissection and mass spectrometry: proteomic tissue profiling of Eudiplozoon nipponicum (Pavel Roudnický) ...... 63

Molecular characterization of gill monogeneans (Monogenea: Dactylogyridae) of African freshwater fishes (Mária Seifertová) ...... 64 The occurrence of the non-native tapeworm Khawia japonensis (Cestoda) in cultured common carp in the Czech Republic confirms its recent expansion in Europe (Tomáš Scholz) ...... 65 Redescription of Sciadocephalus megalodiscus Diesing, 1850, an unusual fish tapeworm (Cestoda: Proteocephalidae) (Tomáš Scholz) ...... 66 A digest of fish tapeworms (Tomáš Scholz) ...... 67 The first record of the invasive Asian fish tapeworm (Schyzocotyle acheilognathi) from an endemic cichlid fish in Madagascar (Tomáš Scholz) ...... 69 An annotated list and molecular data on larvae of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) from freshwater fishes in Africa (Tomáš Scholz) ...... 70 Guide to the Parasites of African Freshwater Fishes: Diversity, Ecology and Research Methods (Tomáš Scholz) ...... 71 Diversity of monogeneans of freshwater fish and coevolution in monogenean-fish systems (Andrea Šimková) ...... 72 New data on cyst ultrastructure of the amoebozoan species Flamella arnhemensis (Tomáš Tyml) ...... 73 Exploring mitogenomics for the phylogeny of African monogeneans (Gyrodactylidae and Dactylogyridae) (Maarten P.M. Vanhove) ...... 75 Eudiplozoon nipponicum (Polyopisthocotylea, Diplozoidae): third monogenean genome at the reach of the hand? (Jiří Vorel) ...... 76 Round goby (Neogobius melanostomus) in the food chain of two Czech rivers.(Lucie Všetíčková) ...... 77 A new species of Aphanoblastella Kritsky, Mendoza-Franco and Scholz, 2000 (Monogenea, Dactylogyridae) parasitic on heptapterid catfish (Siluriformes) in the Neotropical Region (Fabio Hideki Yamada) ...... 78 Study of the tegument, glands and the excretory system of plerocercoids of Dibothriocephalus latus (Cestoda: Diphyllobothriidea) (Aneta Yoneva) ...... 79

List of all publications dedicated to ECIP ...... 80

ECIP Scientific Board

Internal Members

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

Milan Gelnar Andrea Vetešníková Šimková Iva Dyková

Institute of Parasitology, Czech Academy of Sciences, České Budějovice

Tomáš Scholz Astrid Sibylle Holzer (Austria) Aneta Kostadinova (Bulgaria)

Institute of Vertebrate Biology, Czech Academy of Sciences, Brno

Pavel Jurajda Martin Reichard

External Members

Petr Horák (Charles University, Prague, CZ) Kurt Buchmann (University of Kopenhagen, Denmark) Bernd Sures (Universität Duisburg-Essen, Essen, Germany) Carl Smith (University of St Andrews, St. Andrews, UK)

List of attendants Research group of Milan Gelnar – Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno Andrea Bardůnek Milan Gelnar Pavel Roudnický Valigurová Jana Ilgová Eva Řehulková Michal Benovics Tomáš Pakosta Mária Seifertová Kristína Civáňová Martin Kašný Andrea Vetešníková Iva Dyková Šimková Vadym Krasnovyd Kateřina Francová Jiří Vorel Chahrazed Rahmouni Research group of Tomáš Scholz – Institute of Parasitology, Czech Academy of Sciences, České Budějovic Aline Acosta Gabrielle van Beest Carlos A. Mendosa- Palmero Ana Born-Torrijos Roman Kuchta Tomáš Scholz Astrid Holzer Research group of Pavel Jurajda – Institute of Vertebrate Biology, Czech Academy of Sciences, Brno Zdeněk Adámek Pavel Jurajda Martin Reichard Radim Blažek Markéta Ondračková Ludek Šlapanský Michal Janáč Markéta Pravdová Lucie Všetíčková

Programme

Wednesday, November 28th Workshop opening 10:00 – 10:15 Gelnar: Introduction and general programme information

10:15 – 10:30 Gelnar, Scholz & Jurajda: ECIP 2012 – 2018 in numbers

Scientific programme – block I. 10:30 – 11:00 Holzer A.: Research outcomes of the Laboratory of Fish Protistology within the framework of ECIP 11:00 – 11:30 Bardůnek Valigurová A.: Early emerging Apicomplexa from marine invertebrates

11:30 – 12:00 Dyková I.: Key outputs of free-living amoeba research supported by ECIP

12:00 – 12:30 Řehulková E.: Diversity of African monogeneans: Research outlines

12:30 – 14:00 Lunch – Restaurant Nika Scientific programme – block II. 14:00 – 14:30 Kuchta R.: Diversity and phylogenetic relationships of endoparasitic helminths: a brief overview of main achievements 14:30 – 15:00 Vetešníková Šimková A.: Evolutionary ecology of host-parasite interactions

15:00 – 15:30 Kašný M.: From nothing to something…, “omics“ of monogeneans

15:30 – 16:00 Jurajda P. & Reichard M.: Role of host fish in host-parasite interactions

16:00 – 17:00 General Discussion, Workshop closing ceremony and photo of all participants

Social evening 18:00 – 24:00 Dinner (Restaurant Nika)

Abstracts The first part includes abstracts summarizing the work of each individual laboratory. Abstracts in the following section are sorted alphabeticaly according to the first author. Actual or past members of ECIP are presented in bold.

Research outcomes of the Laboratory of Fish Protistology within the framework of ECIP (2012–2017) Astrid S. Holzer1* 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

The Laboratory of Fish Protistology focusses on eukaryotic microorganisms infecting fish, including all aspects of their structure, biology, physiology, life cycles, host-parasite interactions, ecology and especially their phylogeny and evolution. Our main focus is the Myxozoa, extremely reduced fish parasites belonging to the Cnidaria. As part of our work we carry out research into a range of parasite problems which create economic and health consequences for the aquaculture industry. ECIP greatly helped the initial establishment of the Laboratory of Fish Protistology under its newly appointed head (2011) and with the financial support of the project, the laboratory grew considerably, from 7 members to currently 19. Six of the 9 postdocs hired within ECIP are still working in the lab (as contracted scientists or partially or fully contracted postdocs), while all others maintain strong connections. The postdocs have provided the lab with a very wide skill set of methodologies and a complementary knowledge background (transcriptomics and proteomics, immunology, experimental approaches and phylogenetics/-omics). Within the framework of ECIP, funded members of the lab contributed to >30 original papers in peer-reviewed journals, 7 book chapters and numerous conference contributions. 48 % of papers from the lab are in the first and 45 % in the second quartile of the Web of Science citation index, and some articles were published in top journals in their respective field (e.g. PLoS Pathogens, Molecular Ecology). The research performed within ECIP has profoundly developed and driven the investigation in the following fields: Myxozoan type species and poorly characterised myxozoan taxa: A number of genera of myxozoans are scarcely characterized by 18S rDNA gene sequences and the Fish Protistology Lab has contributed to DNA sequencing of more than 70 species in the past 7 years, uncovering an important hidden biodiversity in this group and contributing to the phylogenetic analysis of missing evolutionary links. With this regard, major contributions were made to the Malacosporea, the genera Hoferellus, Sphaeromyxa, Bipteria, Ceratomyxa, Chloromyxum and several other taxa. For the Sphaerospora sensu strico clade, the laboratory members hold the proud record of having characterized all but 3 species (total is 36), with major contributions to a better understanding of the evolutionary history of this group, within the course of ECIP. Due to the experience gained in myxozoan phylogenetics over the last years, the Laboratory of Fish Protistology holds worldwide incomparable expertise in myxozoan phylogenetics, which is actively seeked by researchers from around the world, allowing continuous establishment of new collaborations, which widen the taxa and topics available for phylogenetic research. As an example, the collaboration with the Laboratory of Ichthyoparasitology of the Institute of Marine and Coastal Research (IIMyC), CONICET, Faculty of Exact and Natural Sciences, National University of Mar del Plata, Argentina, which was established in 2013, resulted in important contributions to sequencing unique basal myxozoan taxa from skates and to the detection of myxozoan indicator species for populations studies of Merluccius hubbsi, with two further publications in advanced preparation. Evolution of the Myxozoa: In 2015, based on a phylogenetically basal clustering myxozoan DNA sequence obtained from an evolutionary old, chimaeroid fish, several members of the lab created an interest in the origins of parasitism of the Myxozoa. We performed a first attempt of molecular dating of the myxozoan evolution based on Bipteria vetusta, a myxozoan ‘living fossil’. The results inspired us to examine the history of adaptive radiations in myxozoans and their hosts by determining the degree of congruence between their phylogenies and by timing the emergence of myxozoan lineages in relation to their hosts. Recent genomic analyses suggested a common origin of Polypodium hydriforme, a cnidarian parasite of acipenseriform fishes, and the Myxozoa, and proposed fish as original hosts for both sister lineages. We demonstrate that the Myxozoa emerged long before fish populated Earth and that phylogenetic congruence with their invertebrate hosts is evident down to the most basal branches of the tree, indicating bryozoans and annelids as original hosts and challenging previous evolutionary hypotheses. We were able to provide evidence that, following invertebrate invasion, fish hosts were acquired multiple times, leading to parallel cospeciation patterns in all major phylogenetic lineages. We identified the acquisition of vertebrate hosts that facilitate alternative transmission and dispersion strategies as reason for the distinct success of the Myxozoa, and identify massive host specification-linked parasite diversification events. The novelty of these studies has created an avalanche of inquiries for the datasets used for our analysis and hence are presently fostering further evolutionary and ecological studies.

Ecological studies, fisheries and aquaculture: A number of studies conducted within ECIP focussed on elucidating parasite population genetics, transmission ecology and impacts on fisheries and aquaculture. Investigations into the population structure of clinid fishes and their Ceratomyxa spp. in South African rock pools showed overlapping patterns between parasites and host populations that are very likely affected by oceanographic barriers causing reduced host dispersal and enhancing parasite community differentiation. Another study was performed within the framework of repatriation of Alosa alosa in the river Rhine. We demonstrated that the diversity of SSU rDNA clones of H. alosae was up to four times higher in waters with repatriated hosts and lacked a site-specific signature of SNPs which was clearly observed in long-established populations, indicating that the de novo establishment of myxozoan infections in rivers is slow but of great genetic diversity, likely explained by the introduction of spores from genetically diverse sources, predominantly via straying fish. We also investigated the generalist parasite Cardiocephaloides longicollis as a model for multi-host parasite life cycles in marine habitats. We quantified parasite dispersion and transmission dynamics and the effects of biological changes and anthropogenic impacts on life cycle completion. The study showed a highly significant increase in parasite densities in relation to fishing activities. A study on blood flukes in gills of tuna engorged in Mediterranean netpens elucidated high parasite diversity and demonstrated netpen-based infection of tuna reared in captivity for the first time. Functional studies on parasite virulence and host immune evasion: By identifying genes related to myxozoan virulence and deciphering the immune response of fish to myxozoans as well as immune evasion methods of pathogenic species, we aimed at developing specific anti-myxozoan strategies since there is currently no functioning and legal treatment for myxozoans in fish destined to human consumption. We studied virulence factors in Ceratonova shasta, a serious pathogen in salmonids in the Klamath River system. C. Shasta is a complex of host-specific genotypes of varying virulence. Since myxozoans are not culturable, deep sequencing approaches are notoriously difficult. We developed a two-step bioninformatics pipeline to generate a host-free reference transcriptome of highly virulent genotype IIR for discovery of candidate virulence genes and looked for SNPs in transcriptomes of other genotypes. We determined that motility genes and especially protease-coding genes are strongly associated with virulence in C. shasta and are prime candidates for therapeutic intervention. In the laboratory, we have over the years established Sphaerospora molnari as a model organism for myxozoan research. As a unique achievement worldwide we have developed fully functional in vivo and in vitro models, have produced a genome and stage-specific transcriptomes, which we are presently analysing and mining. Immunological studies and sera antibody trials of samples obtained of infected fish over time, demonstrate that S. molnari raises a strong immunological response in its host, common carp, with massive lymphocyte proliferation and decreased erythrocyte numbers. We demonstrated that carp successfully raise specific antibodies against S. molnari which are fully effective at 35 days post infection, however, the parasite uses immune evasion strategies, i.e. changing antigen patterns, recognizable as different proteins that are bound by host IgM in western blots of parasite extracts, over time.

Future plans Future evolutionary studies in the Laboratory of Fish Protistology will focus on the discovery of new parasite lineages in evolutionary old vertebrate and invertebrate hosts, i.e. sharks and rays as well as sipunculid worms and Haplodrili. This will contribute to a more reliable reconstruction of the evolutionary history of myxozoan parasites. Ecological studies will focus on undetected biodiversity using eDNA and the impact of climate change on parasite proliferation rates. The strongest focus will be on placed on functional transcriptomic and proteomic studies of the presently rather poorly characterized myxozoans and on the development of targeted antiparasitic strategies involving protease inhibitors and vaccines using a combination of antigenic proteins. As a succession of ECIP, we have applied for a large EXPRO project that would produce novel genomic and transcriptomic databases and perform functional research at the forefront of fish parasitology research worldwide.

Acknowledgement These studies were supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

Overview of the work of the Institute of Vertebrate Biology of the Czech Academy of Sciences in the ECIP project Pavel Jurajda1*, Martin Reichard1, Markéta Ondračková1, Michal Janáč1 1Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic *[email protected]

The European Centre of Ichthyoparasitology (ECIP) represented an important project for Fish Biology team of the Institute of Vertebrate Biology. Over the course of the project, 30 people participated as scientific staff and 10 as technical staff. Four scientists (Reichard, Polačik, Jurajda and Ondračková [partial due to maternity leave]) formed the core of team and participated over the whole project. In the first year, we received a number of small investment items, including three microscopes and part of a respirometer, which was used by many team participants over the following project seasons. During the ECIP project, we focused on a number of topics oriented toward host fish of parasites. We investigated the ecology of invasive host fishes and their interaction with native biota. Using round goby (Neogobius melanostomus), we demonstrated that invader impacts are case-specific and unpredictable at local-scales. Our studies further showed that round goby is an attractive host for glochidia of several mussel species, including those of the invasive Chinese pond mussel (Sinanodonta woodiana), revealing the role this invader plays in both parasite spillback (acting as a reservoir for native parasites) and invasional meltdown (facilitating invasion of another non-native species). Brood parasitism was studied in cuckoo catfish (Synodontis multipunctatus) parasitising mouth- brooding cichlid fishes, the only obligate non-avian vertebrate brood parasite. We showed that co-evolutionary history and individual learning in host females contributes to successful host defence. Brood parasitism was also studied using bitterling fishes from Europe and Asia. Using this system, we also investigated components of sexual selection, including the effects and consequences of parasitism on sexual selected traits. As an example, we found that spatial cognitive ability increases the success of parasitic males (an alternative mating strategy in the Eurasian bitterling [Rhodeus amarus]). We studied the ecology of annual fishes in Africa and the Neotropics, including their parasite fauna. We demonstrated complex adaptations of embryonic and adult killifish to their seasonal habitats. We also showed that annual killifish form an important link between aquatic and terrestrial biota and transmit many parasites with complex life cycles from primary to final hosts via trophic interactions. Using Ponto-Caspian gobiids, we demonstrated that decreased infection following introduction contributed to invasion success. While subsequent acquisition of local parasites led to increased parasite richness, susceptibility was buffered by lower parasite fecundity, reducing the importance of non-native hosts as reservoirs. Parasite communities of North-American sunfish (Lepomis gibbosus) mostly comprised co-introduced species (mainly monogeneans), resulting in higher parasite diversity. We documented introduction of both strains of the specific North American digenean Posthodiplostomum centrarchi. Under both experimental and natural conditions, co-introduced parasites did not switch to local fish. Parasite host manipulation was tested using turquoise killifish (Nothobranchius furzeri) and Apatemon trematodes. In replicate field sites in southern Mozambique, a species of the Apatemon genus was located in the brain cavity of infected fish, resulting in conspicuous behavioural changes in the host. Field and experimental observations showed changes in fish habitat use and response to the threat of capture, increasing their capture rate by predators, the definitive trematode hosts. Parasite adaptation to extreme habitats was studied using Nothobranchius parasites of annual killifish inhabiting annually desiccating pools across the African savannah. Parasite abundance was highest in regions with intermediate aridity, while parasite diversity was positively associated with environmental characteristics, fish species diversity and aquatic vegetation. Our results suggest that parasite communities of sympatric Nothobranchius species are similar and dominated by larvae of generalist parasites. As such, Nothobranchius serve as important intermediate or paratenic parasite hosts, with piscivorous birds and predatory fish being the most likely definitive hosts. Parasite infection had a negative effect on the condition of late-hatching naturally infected overwintering European bitterling, resulting in rapid energy depletion. We provided evidence for parasite induced overwintering mortality in juvenile bitterling infected by Diplostomum pseudospathaceum metacercariae. Diplostomum infected males had a larger, though less pronounced, red eyespot; suggesting that non-infected males are better able to attract spawning females. While there was no effect on the number of offspring released or females with developed ovipositors, there was a delay in peak reproduction and offspring

production in infected females. We observed no significant difference in reproductive activity of infected and control males and females during intersexual selection, and a limited effect on female sexual preference. Our recent work has cast doubt on the reliability of traditional fish parasite surveys and we suggested several improvements. Our results suggest that fish should be caught using electrofishing and dissected within three days of capture. Preservation prior to dissection resulted in a loss of information, leading to incomplete quantitative and qualitative data and a lowered ability to determine parasites to species level based on morphology. We published 50 peer-reviewed manuscripts by the end of 2017, with two book chapters and a further 11 manuscripts published or accepted by early 2018. Further manuscripts are in preparation. In addition, the team participated in numerous international conferences. Overall, the ECIP project provided funding for two foreign post-doctorate students and facilitated cooperation with teams from more than ten other countries. Five Masters and four PhD students defended their theses based on data obtained during the project.

Acknowledgement These studies were supported by ECIP (European Centre of Ichthyoparasitology); Centre of Excellence Program of the Czech Science Foundation; Project No. P505/12/G112.

Diversity of African monogeneans: Research outlines (2012−2018) Eva Řehulková1*, Kateřina Francová1, Maria L. Červenka Kičinja1, Mária Seifertová1, Andrea Šimková1, Milan Gelnar1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

During the seven years of the project, most of our research was focused on the diversity (structural and molecular; module 1) and phylogeny (module 2) of monogeneans parasitizing African freshwater fishes. Field trips to Burundi and DR Congo (2013), Sudan (2014), Madagascar (2016), Cameroon (2017) and South Africa (2017) enabled us to sample large numbers of monogeneans representing almost all genera currently known from African fishes. This great collection of monogeneans served as invaluable resources in the preparation of the identification keys to individual genera (i.e. 36 in five families) of these parasites, which form an integral part of the forthcoming publication “A Guide to the Parasites of African Freshwater Fishes” [1]. The uniqueness of the guide lies in the fact that every step in the keys refers to a corresponding figure (representing one species in each genus) for better understanding of identifying feature(s). Although some species were redrawn from the literature, a total of 20 figures were made from specimens in our collection. Furthermore, a catalogue of 482 monogenean species parasitizing African fishes has been compiled as a part of the guide. In addition, our research on the diversity of African monogeneans resulted in the following, more specific outcomes: (1) Diversity of dactylogyrids parasitizing African tetras (Alestidae). As a result of sampling efforts and investigations during the past seven years in eight African countries, an important collection of 40 species (two genera) of monogeneans were found infesting 17 species (six genera) of alestids. Among these parasite species, 33 (ten known, seven newly described, 15 undescribed, and one currently assigned to Afrocleidodiscus) belonged to Annulotrema, and the remaining seven species (four known, three newly described) represented Characidotrema [2,3,4,5]. For the first time, two gene fragments of nuclear ribosomal DNA (18S-ITS1, 28S rDNA) were sequenced for the purposes of molecular species identification and studying the intra/interspecific genetic and phylogenetic relationships in these two genera. The maximum likelihood (ML) and Bayesian analyses (BI) pointed to the monophyletic nature of both Annulotrema (preliminary results) and Characidotrema [5]. In addition, morphological and molecular evaluation of Afrocleidodiscus hydrocynuous from Hydrocynus forskahlii showed that this species should be transferred to Annulotrema based on the morphology of its haptoral armament and its position within cluster of species assigned to the later genus. This result suggests that the host range of Afrocleidodiscus species is restricted only to fishes of the Distichodontidae (Characiformes). (2) Diversity of dactylogyrids parasitizing African catfishes (Siluriformes). During a survey of monogeneans parasitizing catfishes, a total of 11 species belonging to six families (Bagridae, Clariidae, Claroteidae, Malapteruridae, Mochokidae and Schilbeidae) were found to be infected with 27 species of dactylogyrids assigned to six genera: Bagrobdella (4 spp.), Protoancylodiscoides (2 spp.), Quadriacanthus (7 spp.), Schilbetrema (10 spp.), Schilbetrematoides (2 spp.), and Synodontella (2 spp.). The results indicate that our investigation covered a majority of dactylogyrid genera (i.e. all except Birgiellus and Paraquadriacanthus) currently known from catfishes in Africa. Furthermore, a DNA-based approach to identification of these parasites (25 species, 6 genera) was applied for the first time. Nine of the monogenean species found were identified as new to science. Phylogenetic analyses based on 18S-ITS1 and 28S rDNA sequences depicted six Quadriacanthus species, including those described by us (i.e. Q. fornicatus, Q. mandibulatus, Q. pravus, and Q. zuheiri). The observed interspecific genetic relationships among Quadriacanthus spp. from clariids and Q. bagrae from a bagrid host suggest a possible host-switching event in the evolutionary history of the genus [6]. The obvious variety in morphology of ten Schilbetrema spp., now supported by molecular data, and also phylogenetic position of two Schilbetrematoides spp. in relation to Schilbetrema have raised a question on classification of monogeneans infecting schilbeids, which needs to be resolved [7]. (3) Diversity of dactylogyrids parasitizing African/Madagascar cichlids (Cichlidae). As a part of a large- scale study focused on diversity of monogeneans parasitizing cichlids in Lake Tanganyika, descriptions of eight new species of Cichlidogyrus from lamprologine cichlids and proposal of a subgenus within Cichlidogyrus are being finalized together with M. Vanhove [8]. Regarding our investigation focused on monogeneans parasitizing cichlids in Madagascar, the co-introduction of Cichlidogyrus and Scutogyrus with the introduction of their African mainland cichlids into Madagascar was confirmed. However, monogeneans specific to Malagasy cichlids (i.e. species of Insulacleidus) were not found on introduced cichlids co-occurring with native ones [9]. In addition, a

preliminary phylogenetic analysis showed that the endemic genus Insulacleidus is not monophyletic. In collaboration with Malagasy colleagues, a formal taxonomic revision (based on both morphological and molecular data) of monogeneans parasitizing endemic cichlids in Madagascar is being prepared.

Acknowledgements This research was supported by the project ECIP (European Centre of Ichthyoparasitology); Centre of Excellence program of the Czech Science Foundation (Project No. P505/12/G112).

References 1. Řehulková E., Seifertová M., Přikrylová I., Francová K. (2018) A systematic survey of the parasites of freshwater fishes in Africa: Monogenea. In: Scholz T., Vanhove M.P.M., Smit N., Jayasundera Z., Gelnar M. (eds) A guide to the parasites of African freshwater fish. ABC Taxa 18: 185–243. 2. Řehulková E., Musilová N., Gelnar M. (2014) Annulotrema (Monogenea: Dactylogyridae) from Hydrocynus brevis (Günther) (Characiformes: Alestidae) in Senegal, with descriptions of two new species and remarks on Annulotrema pikei. Parasitological Research 113: 3273–3280. 3. Kičinjaová M.L., Blažek R., Gelnar M., Řehulková E. (2015) Annulotrema (Monogenea: Dactylogyridae) from the gills of African tetras (Characiformes: Alestidae) in Lake Turkana, Kenya, with descriptions of four new species and a redescription of A. elongata Paperna and Thurston, 1969. Parasitological Research 114: 4107–4120. 4. Kičinjaová M.L., Barson M., Gelnar M., Řehulková E. (2017) Two new species of Annulotrema (Monogenea: Dactylogyridae) from Hydrocynus vittatus (Characiformes: Alestidae) in Lake Kariba, Zimbabwe. Journal of Helminthology 11: 1–10. 5. Kičinjaová M.L., Seifertová M., Gelnar, M., Řehulková, E. Characidotrema species (Monogenea: Dactylogyridae) from the gills of Brycinus imberi and Brycinus nurse (Characiformes: Alestidae): new records, new species and first insights into the molecular phylogeny of the genus. (submitted) 6. Francová K., Seifertová M., Blažek R., Gelnar M., Mahmoud Z.N., Řehulková E. (2017) Quadriacanthus species (Monogenea: Dactylogyridae) from catfishes (Teleostei: Siluriformes) in eastern Africa: new species, new records and first insights into interspecific generic relationships. Parasites & Vectors 10: 361. 7. Francová K., Seifertová M., Bassock Bayiha E.D., Pariselle A., Bilong Bilong C.F., Gelnar M., Řehulková E. Dactylogyrids parasitizing African schilbeid catfishes (Siluriformes: Schilbeidae) (in preparation) 8. Řehulková E., Šimková A., Pariselle A., Muterezi Bukinga F., Gelnar M., Vanhove M.P.M. A proposal of subgenus within Cichlidogyrus Paperna, 1960 (Monogenea: Dactylogyridae) from lamprologine cichlids (Perciformes: Cichlidae), with description of ten new species. (in preparation) 9. Šimková A., Řehulková E., Rasoloariniaina J.R., Jorissen M.W.P., Scholz T., Faltýnková A., Mašová Š., Vanhove M.P.M. (2018) Transmission of parasites from introduced tilapias: a new threat to endemic Malagasy ichthyofauna. Biological Invasions. https://doi.org/10.1007/s10530-018-1859-0

Diversity and phylogenetic relationships of endoparasitic helminths: a brief overview of main achievements (2012–2018) Tomáš Scholz1*, Aneta Kostadinova1, František Moravec1, Roman Kuchta1, Philippe Vieira Alves1, Anirban Ash1, Daniel Barčák1, Eva Bazsalovicsová1, Ana Born-Torrijos1, Jan Brabec1, Anna Faltýnková1, Simona Georgieva1, David González-Solís1, Jesus S. Hernández-Orts1, Dagmar Jirsová1, Olena Kudlai1, Mikuláš Oros, Miroslava Soldánová1, Aneta Yoneva1 Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

Research was largely focused on the morphology, taxonomy, phylogeny, host-parasite interactions and ecology of endoparasitic helminths, mainly tapeworms, trematodes and nematodes parasitic in freshwater and marine teleosts and freshwater snails (Gastropoda). A high number of new taxa including several new genera have been proposed, many poorly known species redescribed and their phylogenetic relationships assessed using molecular data. Cestoda (Scholz, Kuchta, Alves, Ash, Barčák, Bazsalovicsová, Brabec, Hernández-Orts, Jirsová, Oros and Yoneva): Number of field trips throughout the world were realized to obtain valuable material of tapeworms for subsequent evaluation using methods of integrative taxonomy and phylogenetics. Results were presented in scientific articles and summary of most important achievements was published in 11 chapters in the monograph on the global diversity and interrelations of tapeworms published in 2017 (Caira J.N., Jensen K. (Eds.), 2017: Planetary Biodiversity Inventory (2008–2017): Tapeworms from Vertebrate Bowels of the Earth. University of Kansas, Natural History Museum, Special Publication No. 25, Lawrence, USA, 464 pp.). For example, the relationships of 59 species of the order Bothriocephalidea covering approximately 70% of currently recognised genera were assessed using multi-gene molecular phylogenetic analyses. The order as currently circumscribed, while monophyletic, includes three non-monophyletic and one monophyletic families. The Bothriocephalidae is monophyletic and forms the most derived lineage of the order, comprised of a single freshwater and several marine clades (Brabec et al., 2015). In another important group of fish tapeworms, Proteocephalidea (now part of the Onchoproteocephalidea), a large 28S rDNA-based phylogeny confirmed the limitations of established morphological characters for their classification (de Chambrier et al., 2015: ZooKeys 500: 25–59). The utility of multi-gene molecular phylogenetic analyses was evaluated for use in circumscribing generic boundaries and estimating interrelationships in the caryophyllidean cestodes, which represent an unusual group of tapeworms lacking serially repeated body parts that potentially diverged from the common ancestor of the Eucestoda prior to the evolution of segmentation. Results show that these commonly employed markers do not contain sufficient signal to infer well-supported phylogenetic estimates due to substitution saturation (Brabec et al., 2012). Trematoda (Kostadinova, Born-Torrijos, Faltýnková, Georgieva, Kudlai and Soldánová): Several studies addressing integrated molecular and morphological approaches to the species diversity of diplostomid trematodes, whose metacercariae may be pathogenic for fish were carried out using isolates of larval and adult stages from Europe, Asia and Africa. Extensive dataset of newly generated sequences of several molecular markers has been generated as a baseline for future ecological and evolutionary studies of trematodes and mainly their larval stages, which are unidentifiable based on their morphology only. The first comprehensive phylogeny for the superfamily Echinostomatoidea was published based on lsrRNA gene for 80 species, representing eight families and 40 genera. This resulted in a number of systematic and nomenclatural changes consistent with the phylogenetic estimates (Tkach et al., 2016). The biodiversity of trematodes in their intermediate mollusc and fish hosts in the freshwater environment in Europe was assessed based on two novel databases containing 2,380 and 8,202 host-parasite- locality records for trematode parasites of molluscs and fishes, respectively. Analyses revealed rich trematode faunas in snails (171 species of 89 genera and 35 families; of these 23 and 40 species utilise freshwater fishes as definitive and second intermediate hosts, respectively). The most frequently recorded species are Diplostomum spathaceum, D. pseudospathaceum, Tylodelphys clavata and Posthodiplostomum cuticola (Faltýnková et al., 2016: Systematic Parasitology 93: 283–293). The biodiversity patterns of digeneans of fishes in the Mediterranean were analysed based on an updated database comprising 302 digenean species allocated to 146 genera in 29 families from 192 fish species of 76 families. Results indicated high discovery rates (2.4 new species per year) due to the application of combined morphological and molecular methods, which promise more reliable estimates of digenean diversity in this well-

studied region. Host-parasite lists for 192 species of fishes in the Mediterranean comprising 890 host-parasite associations were also provided (Pérez-del-Olmo et al., 2016: Systematic Parasitology 93: 249–256). Nematoda (Moravec and González-Solís): Investigations were largely oriented to studies on the morphology, taxonomy, host-parasite relationships and distribution of nematodes parasitic mainly in marine and freshwater fishes over the world. The great majority of studies were those on philometrid nematodes (Philometridae), a species-rich group of very important, but so far little-known. Detailed studies of these parasites revealed a total of 50 previously unknown species and resulted in the erection of three new genera. An additional 37 new nematode species including two new genera were discovered and described from freshwater and marine fishes. Moreover, these studies enabled to redescribe in detail many insufficiently known species and to solve up a number of taxonomic problems concerning these parasites. The above results were presented in 76 papers published in impacted international parasitological journals and a monograph dealing with nematode parasites of African inland fishes (Moravec, 2018). Until the end of October 2018, the results were presented in 189 papers published in impacted international parasitological journals (sum of impact factors = 279.51), two monographs (Moravec, 2018; Scholz et al., 2018) and 21 chapters in books. Eleven articles were published in the journals of the first quartile (Q1 – see the list below) and another 29 in the second quartile (Q2). During duration of the project, several young researchers (postdocs) and graduate students from the following countries abroad were temporarily hired: Brazil (1 PhD completed), India (1 PhD thesis), Mexico, Slovakia (1 PhD thesis), Spain and Ukraine. They contributed significantly to fulfilling the objectives of the project and their participation in the project represented an important stimulus for their personal development and career advancement.

Acknowledgement Generous financial support of the Czech Science Foundation (project No. P505/12/G112) is highly appreciated.

Selected publications on endohelminths (2012–2018)

Monographs: 1. Moravec F. (2013) Parasitic Nematodes of Freshwater Fishes of Europe. Revised Second Edition. Academia, Praha, 601 pp. 2. Moravec F. (2018) Parasitic Nematodes of Freshwater Fishes of Africa. Academia, Prague (in press). 3. Scholz T., Vanhove M.P.M., Smit N., Jayasundera Z., Gelnar M. (Editors) (2018) A Guide to the Parasites of African Freshwater Fishes. ABC Taxa, CEBioS, Royal Belgian Institute of Natural Sciences, No. 18, 421 pp.

Book chapters: 1. Kuchta R. (10 chapters), Scholz T. (10 chapters), Brabec J. (1 chapter): 11 chapters in J.N. Caira and K. Jensen (Eds.), Planetary Biodiversity Inventory (2008–2017): Tapeworms from Vertebrate Bowels of the Earth. University of Kansas, Natural History Museum, Special Publication No. 25, pp. 1–20, 21–28, 29– 45, 47–64, 167–189, 191–199, 201–206, 243–250, 251–277, 349–356 and 357–370 (a total of 158 pages).

Scientific articles (Q1) 1. Ash A., Scholz T., de Chambrier A., Brabec J., Oros M., Kar P.K., Chavan S.P., Mariaux J. (2012) Revision of Gangesia (Cestoda: Proteocephalidea) in the Indomalayan region: morphology, molecules and surface ultrastructure. PLoS ONE 7: e46421. [IF = 3.730] 2. Brabec J., Scholz T., Kraľová-Hromadová I., Bazsalovicsová E., Olson P.D. (2012) Substitution saturation and nuclear paralogs of commonly employed phylogenetic markers in the Caryophyllidea, an unusual group of non-segmented tapeworms (Platyhelminthes). International Journal for Parasitology 42: 259– 267. [IF = 3.637] 3. Chibwana F.D., Blasco-Costa I., Georgieva S., Hosea K. M., Nkwengulila G., Scholz T., Kostadinova A. (2013) A first insight into the barcodes for African diplostomids (Digenea: Diplostomidae): brain parasites in Clarias gariepinus (Siluriformes: Clariidae). Infection, Genetics and Evolution 17: 62–70. [IF = 3.264] 4. Georgieva S., Soldánová M., Pérez-del-Olmo A., Dangel D.R., Sitko J., Sures B., Kostadinova A. (2013) Molecular prospecting for European Diplostomum (Digenea: Diplostomidae) reveals cryptic diversity. International Journal for Parasitology 43: 57–72. [IF = 3.404]

5. Blasco-Costa I., Faltýnková A., Georgieva S., Skirnisson K., Scholz T., Kostadinova A. (2014) Fish pathogens near the Arctic Circle: molecular, morphological and ecological evidence for unexpected diversity of Diplostomum (Digenea: Diplostomidae) in Iceland. International Journal for Parasitology 44: 703–715. [IF = 3.872] 6. Pérez-del-Olmo A., Georgieva S., Pula H., Kostadinova A. (2014) Molecular and morphological evidence for three species of Diplostomum (Digenea: Diplostomidae), parasites of fishes and fish-eating birds in Spain. Parasites & Vectors 7: 502. [IF = 3.430] 7. Brabec J., Waeschenbach A., Scholz T., Littlewood D.T.J., Kuchta R. (2015) Molecular phylogeny of the Bothriocephalidea (Cestoda): molecular data challenge morphology-based classification. International Journal for Parasitology 45: 761–771. [IF = 3.234] 8. Mendoza-Palmero C.A., Blasco-Costa I., Scholz T. (2015) Molecular phylogeny of Neotropical monogeneans (Platyhelminthes: Monogenea) from catfishes (Siluriformes). Parasites & Vectors 8: 164. [IF = 3.234] 9. Tkach V.V., Kudlai O., Kostadinova A. (2016) Molecular phylogeny and systematics of the Echinostomatoidea Looss, 1899 (Platyhelminthes: Digenea). International Journal for Parasitology 46: 171–185. [IF = 3.370] 10. Jirsová D., Štefka J., Jirků M. (2017) Discordant population histories of host and its parasite: a role for ecological permeability of extreme environment? PLoS ONE 12: e0175286. [IF = 2.766]

Evolutionary ecology of host-parasite interactions Andrea Šimková1* 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

The host-parasite interactions represent one of the most serious topics in evolutionary ecology. In host-parasite interactions module we basically focused on the diversity and phylogeny of selected groups of monogeneans parasitizing the freshwater fish in order to investigate the patterns and processes of host-parasite coevolution and host biogeographical patterns revealed by host specific monogeneans. We also investigated the potential role of parasites in the evolution of host immune system. Dactylogyrus parasites specific to cyprinids represented the first host-parasite system studied. Our studies revealed the high morphological diversity and the extremely high genetic diversity of Dactylogyrus parasitizing Euro-Mediterranean cyprinids. The cospeciation was detected only at the level of the highest taxonomical level of cyprinids and host switch was recognized as the main frequent coevolutionary event of Dactylogyrus speciation and diversification in endemic areas. Using Dactylogyrus parasitizing cyprinids from the Balkans we showed that the evolution of these cyprinid specific monogeneans has been influenced by the historical dispersion and distribution of their cyprinid hosts, but also by recent contacts of endemic and non- native cyprinid species. Cophylogenetic analyses performed using North African cyprinids revealed also the cospeciation at the high taxonomic level of cyprinids (cyprinid lineages), and host switch was documented as the major event in morphologically and ecologically similar Luciobarbus. We also focused on generalist Dactylogyrus forms by selecting D. vistulae (a species parasitizing a wide range of cyprinid species) and investigating population structure of this species across widespread and endemic cyprinid species in Europe. Overall, no considerable differences in genetic variability were revealed between populations from widely distributed cyprinids in central Europe and endemic cyprinids exhibiting restricted distribution in Mediterranean area. However, low variability in D. vistulae populations was detected particularly in endemic cyprinids. Genetic variability of D. vistulae populations might, therefore, reflect genetic variability of their hosts. Cichlidogyrus parasites specific to cichlids represented the second host-parasite system studied. Our studies revealed the high morphological and molecular diversity of Cichlidogyrus parasitizing highly diversified cichlids from the Lake Tanganyika. Our phylogenetic analyses revealed the potential role of host-specific Cichlidogyrus to elucidate the history and biogeography of cichlids living in the Lake Tanganyika. Cichlids exhibit interesting disjunctive word wide distribution and harbor different monogeneans (belonging to different genera) in different biogeographical areas. We focused on the distribution and phylogeny of ectoparasitic monogeneans host specific to cichlids living recently in allopatry and we demonstrated the independent evolution of host specific fauna following historical dispersion of cichlids. In accordance with molecular phylogenetic studies focused on cichlid fish, we showed the polyphyletic origin of monogeneans parasitizing Malagasy cichlids. The analyses of ecological data showed that Malagasy cichlids are susceptible to the infection of host specific monogeneans originated in African mainland (Cichlidogyrus and Scutogyrus genera). On the other hand, we showed the monophyletic group of endoparasitic monogeneans of cichlids living in Africa and South Asia suggesting the potential historical cichlid crossing of the sea together with endoparasitic monogeneans but the loss of ectoparasitic fauna intolerant to salinity. Finally, diversity and phylogeny of host specific parasites of cichlids living in South America were investigated suggesting that the phylogeny of host specific gill monogeneans follows the phylogeny of diverse cichlid lineages in this continent. Anacanthorus parasitizing Characiformes, especially Bryconidae, Characidae and Serrasalmidae represent the third host-parasite system studied. We focused on Anacanthorus species infecting Serrasalmidae. The phylogenetic analyses indicated that the relationships among Anacanthorus species reflect the relationships between the lineages of the serrasalmid hosts: Anacanthorus clustered following their host specificity to following serrasalmid lineages: Piaractus, member of the “pacus” lineage, the “Myleus-like pacus” and the lineage of the “true piranhas”. The role of parasite in evolution of immune systems was investigated using several host systems. Major histocompatibility system represent the important part of vertebrate immune adaptive system. Parasite- mediated selection (balancing selection) and reproductive mechanisms based on mating preferences are the most often cited mechanisms maintaining the extensive MHC polymorphism in fish. Parasite-mediated selection driving the MHC class IIB diversity was investigated in wild populations of European chub (Squalius cephalus). The populations showed similar patterns of variation in parasites, immunogenetic and neutral markers, which strongly corresponds to their phylogeographic structure previously inferred from microsatellites. The analyses revealed that populations with dissimilar MHC allelic profiles were

geographically distant populations with significantly different diversity in microsatellites and a dissimilar composition of parasite communities. The metazoan parasite load at the level of individuals was influenced by the diversity of DAB alleles as well as by the diversity of neutral genetic markers and host traits reflecting condition and immunocompetence. The multivariate co-inertia analysis showed specific associations between DAB alleles and parasite species. In this study we suggest that parasite-mediated selection is not the only mechanism affecting MHC diversity in European chub; we strongly support the role of neutral processes as the main driver of DAB diversity across populations of wild living fish. The variability of exon 2 of DAB genes 2 (i.e. MHC IIB genes) were studied in the populations of invasive Barbus barbus, endemic Barbus meridionalis and their respective F1 hybrids from the hybrid zones in Southern France. Our results indicate that positive selection significantly influences the variability of DAB-like alleles in Barbus populations. However, similar pattern of genetic structure based on DAB genes and microsatellites, and the evidence of the alleles shared between species suggest both the effects of neutral selection and trans-species polymorphism on MHC diversity in B. barbus and B. meridionalis populations. The recombination, duplication and gene flow show to play no important role in shaping variability of MHC genes. Low levels of variability of DAB-like alleles reported in B. meridionalis might be connected with poor immune system of fish since high metazoan parasite infection B. meridionalis was previously reported. The variability of MHC genes was also studied in the natural hybrids of phylogenetically distant species – common bream (Abramis brama) and roach (Rutilus rutilus) species with different morphology, ecology and evolutionary divergence. Each species exhibit the specific parasite fauna especially concerning the monogeneans. The frequency of F1 hybrids was very low in nature. The study of parasite load in pure species and F1 hybrids confirmed the scenario of hybrid resistance i.e. level of parasite infection in hybrids was lower than level of parasite infection in pure species. Hybrids harbor the specific parasites of both species. However, they were more parasitized by monogeneans specific to roach. The analyses of MHC showed that parental species differ in the MHC variability. In addition, MHC variability was lower in hybrids comparing to each of parental species. F1 hybrids shared alleles of MHC genes more with roach than with common bream, which may be related to the higher abundance of roach specific parasites in hybrids. The role of parasite in the evolution of diploid-polyploid complex exhibiting the sexual and asexual reproduction was investigated in the coexisting sexual and gynogenetic forms of gibel carp, Carassius gibelio (Cyprinidae). The sampled gynogenetic form was composed of specimens expressing the common MHC genotypes and of specimens expressing rare genotypes. In contrast, the sexual form of gibel carp exhibited a wide range of MHC genotypes (and expressed a wide range of DAB-like alleles), which is in line with the prediction of sexually-mediated selection increasing MHC diversity. The analysis of the MHC diversity documented a change in the most common genotypes of the gynogenic form over time. Changes in MHC genotype frequencies could by likely explained by temporal variations in parasite load based on the frequency-dependent selection. Finally, toll-like receptors (TLRs), a group of transmembrane protein receptors, playing a crucial role in innate immunity response to pathogens was also investigated in Abramis brama x Rutilus rutilus hybrid system. We compared TLRs genes expression patterns in different tissues (spleen, head kidney and gills) in parental species and their F1 hybrids. For the fish sampled, considerably higher abundance of monogenean parasites was revealed in A. brama in comparison with R. rutilus and hybrids. At the same time, significantly higher TLR21 expression was observed in A. brama than in remaining fish groups. Significant effect of monogenean abundance and fish group (i.e. A. brama, R. rutilus and hybrids) on TLR21 expression was subsequently statistically supported. No effect of monogenean abundance was revealed on TLR21 expression in remaining tissues and TLR2 expression. However, in A. brama significant negative correlation was observed between the abundance of strict specific Dactylogyrus zandti and expression of TLR21 in gills.

Acknowledgement This study was largely supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112. I acknowledge all colleagues participated in the above mentioned studies and especially the following students Michal Benovics, Imane Rahmouni, Juliana Moreira, Lenka Gettová, Adam Potrok, Tomáš Pakosta and Vadym Krasnovyd working on topics summarized in this abstract.

Early emerging Apicomplexa from marine invertebrates Andrea Valigurová1*, Andrei Diakin1, Magdaléna Kováčiková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

Apicomplexa Levine, 1980, emend. Adl et al. 2012 represent one of the most successful groups of eukaryotic unicellular parasites that infect almost all known phyla of multicellular organisms. They comprise numerous etiologic agents of significant human diseases, representing a major world health problem with a considerable impact on the global economy (e.g. malaria, toxoplasmosis). In contrast to vertebrate pathogens, apicomplexans restricted to invertebrates are considered of no economic or medical significance and despite their enormous diversity remain poorly understood. They are, however, important from an evolutionary perspective because of their basal phylogenetic position. It is generally accepted that ancestral apicomplexans parasitised marine annelids, and their radiation and adaptation to the parasitic life style took place before the era of vertebrates. First they spread to other marine invertebrates, then to freshwater and terrestrial invertebrates and, finally, vertebrates. Early emerging Apicomplexa exhibit an enormous diversity in cell dimensions and architecture, depending on their surrounding environment and parasitic strategy, and seem to be a perfect example of a coevolution between a group of parasites and their hosts. We investigated fauna of early dispersed apicomplexan branches from invertebrates collected at the White Sea (Russia), English Channel near the Station Biologique de Roscoff (France) and along the shore of James Ross Island, Weddell Sea near the J.G. Mendel Station (Antarctica), with emphasis on protococcidia, agamococcidia, blastogregarines, archigregarines, and eugregarines. Our research focused on three main topics: biodiversity, host-parasite interactions and motility in model parasites. Biodiversity. Until recent, blastogregarines were poorly studied parasites of polychaetes with uncertain taxonomic affiliation. They superficially resemble gregarines, but lack the stage of syzygy and gametocyst in their life cycle. Previous studies considered them highly modified gregarines, an intermediate lineage between gregarines and coccidia, or an isolated group of eukaryotes altogether [1]. Blastogregarines exhibit a permanent multinuclearity and gametogenesis by means of budding, distinguishing them considerably from the other Apicomplexa. We studied Chattonaria mesnili (Chatton et Dehorne, 1929) from the intestine of polychaete Orbinia latreillii (Audouin et Milne-Edwards, 1833) and the type species, Siedleckia nematoides Caullery et Mesnil, 1898 from the intestine of polychaete Scoloplos armiger (Müller, 1776) [1,2]. Our molecular phylogenetic analyses showed blastogregarines as an independent, early diverging lineage of Apicomplexa. Both species exhibit the gregarine (longitudinally folded or smooth pellicle, the persistence of mucron and apical complex during the larger part of lifecycle, oocysts with free sporozoites) and coccidian (gametes are associated with two kinetosomes, absent gametocyst, male and female gametes of different size, microgametes with two flagella) morphological features. The traits shared with Selenidium archigregarines, i.e. a distinctive tegument structure and myzocytosis via a well-developed apical complex, likely represent the ancestral states of the corresponding cell structures for Apicomplexa [1,2]. Archigregarines occur only in marine environments and are expected to be the most recent ancestor of gregarines. By their phylogenetic position, they are an important lineage to understand the functional transition that occurred between free-living flagellated predators to obligatory parasites in Apicomplexa. We investigated the type species, Selenidium pendula Giard, 1884 from the intestine of marine polychaete Scolelepis squamata (Müller, 1806) [3]. We described for the first time the stage of syzygy and early gamogony at the ultrastructural level. Analysis of SSU rDNA sequences showed S. pendula belonging to a monophyletic lineage that includes several other related species (parasites of Sedentaria), sharing similar biological characters [3]. The other two species investigated were newly described species from the intestine of marine polychaetes, S. pygospionis Paskerova et al., 2018 from Pygospio elegans Claparède, 1863 and S. pherusae Paskerova et al, 2018 from Pherusa plumosa (Müller, 1776) [4,5]. Ultrastructure and phylogeny was studied in three septate eugregarines from crustaceans: Cephaloidophora cf. communis Mawrodiadi, 1908 from the intestine of marine barnacle Balanus balanus Linnaeus, 1758 [6]; Heliospora cf. longissima (von Siebold in Kölliker, 1848) Goodrich, 1949 from freshwater amphipods, Eulimnogammarus verrucosus Gerstfeldt, 1858 and E. vittatus Dybowski, 1874 [6], and Ganymedes yurii Diakin, Wakeman et Valigurová, 2016 from the intestine of marine amphipod Gondogeneia sp. [7]. Urospora ovalis Dogiel, 1910 and U. travisiae Dogiel, 1910 from the body cavity of marine polychaete Travisia forbesii Johnston, 1840 were studied with an emphasis on general morphology and phylogenetic position [8]. A robust ultrastructural and molecular phylogenetic study was performed on the marine eugregarine Ancora sagittata (Leuckart, 1860) Labbé, 1899 from the intestine of polychaete Capitella capitata Fabricius, 1780 [9].

Host-parasite interactions. Apicomplexan zoite exhibits a high cell polarity as it has an anterior pole equipped with a unique invasion apparatus, the apical complex. This apparatus can be also found in other Myzozoa - a group comprising apicomplexans, dinoflagellates, and several lineages of free-living predatory or parasitic flagellates that employ a myzocytosis-based mode of feeding (= predator/parasite attaches to the prey/target cell and sucks out or feeds permanently on its cytoplasm via specialized organelles). We expect that the evolution of the Apicomplexa progressed from myzocytotic predation to myzocytotic extracellular parasitism, as exhibited by gregarines and cryptosporidia, and to intracellular parasitism typical for coccidia. The attachment apparatus of blastogregarines and gregarines evolved on the apical end of sporozoite, and demonstrates an enormous diversity in its architecture: from a mucron in blastogregarines and archigregarines [1-4], through an more advanced mucron-like organelle in aseptate eugregarines losing the apical complex and strengthening its attaching function, simple [6,9,12] or complicated epimerite of septate eugregarines, and finally modified protomerite in septate eugregarines. Protococcidian Eleutheroschizon duboscqi Brasil, 1906 (the type species) develops epicellularly attached to the intestinal epithelium of the marine polychaete S. armiger. The formation of parasitophorous sac is very similar to that in cryptosporidia, while the parasite itself along with its attachment site resembles an invading gregarine [10]. Agamococcidian Rhytidocystis sp. from marine polychaete T. forbesii develops intracellularly within enterocytes, without the presence of parasitophorous vacuole [11]. Motility. Basal lineages differ from the other Apicomplexa in that their trophozoite and gamont stages are often motile at least to some degree, and their locomotion differs from the substrate-dependent, actin/myosin-based gliding motility in motile apicomplexan zoites (the glideosome concept). They appear to use several mechanisms of cell motility, correlating with various modifications of cell cortex. Different modes of motility could represent specific adaptation to parasitism in different environments within their hosts. Both the Selenidium archigregarines and blastogregarines move independently on a solid substrate without any signs of gliding motility [1-5]. The subpellicular microtubules organised in several layers seem to be the leading motor structures in their motility [2,5]. While archigregarines are typical by a very active pendular movement [3-5], the motility of blastogregarines rather resembles a sequence of pendular, twisting, undulation, and sometimes spasmodic movements [1,2]. Eugregarines parasitising intestines of crustaceans and polychaetes move by a linear gliding [6,7,9,12], whereas eugregarines from coelom exhibit peristaltic or metabolic movements [8]. Our data indicate that the eugregarine gliding is facilitated by epicytic folds (bearing at their tops the 12-nm filaments responsible for the gregarine gliding path) and supported by a secretion of mucopolysaccharides [12].

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

References 1. Simdyanov T.G., Paskerova G.G., Valigurová A., Diakin A., Kováčiková M., Schrével J., Guillou L., Dobrovolskij A.A., Aleoshin V.V. (2018) First Ultrastructural and molecular phylogenetic evidence from the blastogregarines, an early branching lineage of plesiomorphic Apicomplexa. Protist 169: 697–726 2. Valigurová A., Vaškovicová N., Diakin A., Paskerova G.G., Simdyanov T.G., Kováčiková M. (2017) Motility in blastogregarines (Apicomplexa): Native and drug-induced organisation of Siedleckia nematoides cytoskeletal elements. PLOS ONE 12(6): e0179709. 3. Schrével J., Valigurová A., Prensier G., Chambouvet A., Florent I., Guillou L. (2016) Ultrastructure of Selenidium pendula, the type species of archigregarines, and phylogenetic relations to other marine Apicomplexa. Protist 167: 339–368. 4. Paskerova G.G., Miroliubova T.S., Diakin A., Kováčiková M., Valigurová A., Guillou L., Aleoshin V.V., Simdyanov T.G. (2018) Fine structure and molecular phylogenetic position of two marine gregarines, Selenidium pygospionis sp. n. and S. pherusae sp. n., with notes on the phylogeny of Archigregarinida (Apicomplexa). Protist (in press). 5. Kováčiková M., Paskerova G.G., Diakin A., Valigurová A. (2017) Cytoskeletal elements and motility in the archigregarine Selenidium sp.: observations on native and drug-treated parasites. In: Book of Abstracts 15th International Congress of Protistology. July 7-August 4, 2017 Prague, Czech Republic. p. 113. 6. Simdyanov T.G., Diakin A.Y., Aleoshin V.V. (2015) Ultrastructure and 28S rDNA Phylogeny of two gregarines: Cephaloidophora cf. communis and Heliospora cf. longissima with remarks on gregarine morphology and phylogenetic analysis. Acta Protozoologica 54(3): 241–263. 7. Diakin A., Wakeman K.C., Valigurová A. (2016) Description of Ganymedes yurii sp. n. (Ganymedidae), a new gregarine species from the Antarctic amphipod Gondogeneia sp. (Crustacea). Journal of Eukaryotic Microbiology 64(1): 56–66.

8. Diakin A., Paskerova G.G., Simdyanov T.G., Aleoshin V.V., Valigurová A. (2016) Morphology and molecular phylogeny of coelomic gregarines (Apicomplexa) with different types of motility: Urospora ovalis and U. travisiae from the polychaete Travisia forbesii. Protist 167: 279–301. 9. Simdyanov T.G., Guillou L., Diakin A.Y., Mikhailov K.V., Schrével J., Aleoshin V.V. (2017) A new view on the morphology and phylogeny of eugregarines suggested by the evidence from the gregarine Ancora sagittata (Leuckart, 1860) Labbé, 1899 (Apicomplexa: Eugregarinida). PeerJ 5: e3354. 10. Valigurová A., Paskerova G.G., Diakin A., Kováčiková M., Simdyanov T.G. (2015) Protococcidian Eleutheroschizon duboscqi, an unusual apicomplexan interconnecting gregarines and cryptosporidia. PLoS ONE 10(4): e0125063. 11. Diakin A., Valigurová A. (2014) Development of new species of agamococcidian Rhytidocystis sp. from Travisia forbesii. In: A variety of interactions in the marine environment. Abstracts volume from 49th European Marine Biology Symposium, Zoological Institute Russian Academy of Sciences. September 8– 12, 2014 St. Petersburg, Russia. pp. 83–84. 12. Kováčiková M., Simdyanov T.G., Diakin A., Valigurová A. (2017) Structures related to attachment and motility in the marine eugregarine Cephaloidophora cf. communis (Apicomplexa). European Journal of Protistology 59: 1–13.

Fig 1. Blastogregarine Siedleckia nematoides, immunofluorescent labelling of α-tubulin (green), myosin (red) and nuclei (blue) (CLSM). Fig 2. Archigregarine Selenidium pygospionis (SEM). Fig 3. Macrogamont of protococcidian Eleutheroschizon duboscqi attached to the host intestinal epithelium (TEM).

From nothing to something…, “omics” of monogeneans Jiří Vorel1, Jana Ilgová1, Pavel Roudnický1, Libor Mikeš2, Hana Dvořáková2, Lucie Jedličková2, Dáša Jirsová1, Hynek Strnad3, Roman Leontovyč2, Ewa Dzika4, Božena Koubková1, Lukáš Vetešník5, David Potěšil6, Zbyněk Zdráhal6,7, Marie Jankůjová7, Jan Oppelt7, Pavel Jurajda5, Milan Gelnar1, Martin Kašný1,2* 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic; 3Genomics and Bioinformatics, Institute of Molecular Genetics, Czech Academy of Sciences, Prague, Czech Republic; 4Faculty of Medical Science, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland; 5Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic; 6CEITEC – Proteomics Core Facility, Masaryk University, Brno, Czech Republic; 7National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

The ectoparasitic worms from the group Monogenea are primarily infecting the (semi-)aquatic vertebrates, predominantly fishes. Members of this group are not of direct human health concern, but a number of species are known to cause huge economic losses in the aquaculture and fishing industries, e.g. the “salmon killer” Gyrodactylus salaris, in Norwegian economy ca 50 mil. US$/year. During the last decade the number of flatworms, such as trematodes and cestodes, sampled at a proteome, transcriptome and genome level significantly increased. In contrast, the monogeneans are currently still represented by only one published draft genome for G. salaris, by one not published draft genome Protopolystoma xenopodis (on the list of the “50 Helminth genome project initiative”) and one transcriptome dataset for Neobenedenia melleni. The lack of data and information at this field induced our experimental effort to generate relevant sequential data source related to Monogenea. We used the experimental organisms - Eudiplozoon nipponicum (Diplozoidae, Heteronchoinea); a blood-feeding monogenean inhabiting gills of common carp (Cyprinus carpio) and during last 7 years, in the frame of the project ECIP, we applied the high throughput sequencing methods (454/Roche, MiSeq/HiSeq) which enabled us to yield robust trancriptomic/genomic data and perform the comprehensive mass spectrometry analysis (Orbitrap-Elite MS System) of excretory-secretory products isolated from the adult E. nipponicum worms. These data enabled us for the first time within the group Monogenea to elucidate the properties of selected key genes/proteins playing the important roles in the biology of these interesting organisms.

The selected functional protein molecules have been molecularly/biochemically/immunologically characterized. Genome: Whole genome analysis was performed by a combination of sequencing techniques 454/Roche, Illumina Hiseq and Miseq and Oxford Nanopore sequencing and in total 166,429,733 reads (about 29,5 Gb) were generated. According to estimated genome size 1.5 Gb evaluated by flow cytometry and k-mer counting, this number is corresponding to potential 20x coverage. Our first draft of genome assembly produced a very fragmented genome about total size 1,1 Gb. Therefore, we started to work on the second draft of genome assembly, improved by very long nanopore reads (2,755,771 reads, 8,0 Gb). After the first assembly steps, we were able to reconstruct the entire

E. nipponicum mitochondrial genome with the length of 16,727 bases encoding 36 genes. Transcriptome: During the E. nipponicum analysis of transcriptome 324,941 454/Roche and 149,697,864 Illumina raw reads were produced and assembled in 37,062 transcripts and 19,644 (53.0 %) of them were annotated (based on the homology with at least one record in used public sequence databases). 18,657 (50.3 %) were assigned to sequences of parasitic flatworms (TrEMBL, taxon ID: 6157). On the base of annotation results it was revealed e.g. that metallopeptidases are the most expressed group of proteins with catalytic activity (2,304 transcripts; 6.2 %). Performing the gene function analysis (GO terms) based on the homology with at least one record in used public sequence databases 11,402 (30.8 %) transcripts described by at least one GO term were determined. Among the most abundant pathways were “the classic biochemical pathways”, such as protein binding (GO:0005515, n = 1,556 transcripts), ATP binding (GO:0005524, n = 860 trancripts) and nucleic acid binding (GO:0003676, n = 733 transcripts). The group of transcripts revealed at position 19 - GO term proteolysis (GO:0006508) involved functionally important enzymes, e.g. cathepsins L, B and D which properties were further investigated (see below). Proteome: The excretory-secretory products from 100 worms were isolated, concentrated, purified and analysed using RSLCnano system connected to Orbitrap Elite hybrid spectrometer. The analysis of the mass spectrometric raw data files was carried out using the Proteome Discoverer software with in-house Mascot search engine utilisation. MS/MS ion searches were done at first against modified cRAP database containing protein contaminants like keratin, trypsin etc. MS/MS spectra assigned to any cRAP protein peptide with Mascot ion score >30 were excluded from the next database searches. In-house protein database for E. nipponicum concatenated following databases; E. nipponicum transcriptome (37,062 protein sequences) and C. carpio proteins derivate from genome (63,928 sequences, NCBI Genome ID: 10839). In ESP 727 proteins were identified, among them also the relevant functional molecules know for other platyhelminths, e.g. peptidases (cathepsin L, B and D, aspartic peptidase, metalopetidase MEP1, dipeptidyl amino peptidase) – some of them are probably involved in digestion of blood and peptidase inhibitors (stefin, serpin, kunitz, kazal inhibitor) – enabling the regulation of host´s physiological

23 processes such as blood coagulation or inflammation. Further we developed a laser dissection/mass spectrometry technique enabling to analyse the proteome of selected E. nipponicum tissue and thus to localise the particular protein molecule. Until now we have identified 405 proteins in worm body parenchyma (including also gut) and 131 proteins specific for tegument. Among them, some already described functional molecules have been found, e.g. cathepsin L, serpin. Particular protein molecules: During the experimental sequential work we obtained very interesting results concerning bioactive molecules of E. nipponicum. We found that cysteine and aspartic endopeptidases (cathepsins L, B and D) take significant part in digestion of hemoglobin in E. nipponicum and peptidase inhibitors (serpin, stefin) are able to regulate the mentioned proteolytic enzymes. The selected protein molecules – peptidases/peptidase inhibitors were produced in recombinant form and molecularly, biochemically, immunologically characterized. The digestion of blood-feeding Monogenea was originally believed to take place in specialized cells lining the intestinal epithelium (also called hematin cells). However, the detection of vast amount of peptidases in the ESP (cathepsin L, B and D, aspartic peptidases) supports alternative hypothesis of initial breakdown of hemoglobin outside of these cells, possibly within the gut lumen or in the host’s capillaries. Besides controlling endogenous processes, e.g., digestion of the parasite, the E. nipponicum peptidase inhibitors may participate in the modulation of the host´s immune system. We focused on cysteine peptidase inhibitor (EnStef) and characterized its biochemical properties and possible function in regulation of proteolysis during food processing. In addition to containing papain binding domain, EnStef possesses a unique motif important for inhibition of legumains (group of peptidases crucial in immune defense of the host). Surprisingly, no legumain activity was detected in ESP or crude worm extract and hence it is tempting to hypothesize that secreted stefin with the ability to inhibit legumain possibly targets asparaginyl peptidases of the host origin. Further evidences for this statement as well as the determination of specific role of stefin are subjected to running study. However, we already verified immunosuppressive effect of recombinant cystatin on expression of TNF-alpha and IL-10 by macrophages. Among the serine peptidase inhibitors, the transcripts of E. nipponicum serine peptidase inhibitor - serpin (EnSerp1) were revealed. Using mass spectrometric techniques and immunoblot EnSerp1 in E. nipponicum ESP was confirmed. Fluorometric inhibition assays showed the rEnSerp1 ability to partially inhibit four serine peptidases playing a role in host-parasite interaction – digestion (trypsin), regulation of blood coagulation (factor Xa, plasmin) or tempering the inflammation (kallikrein). Due to properties mentioned above and presence of the serpin in ESP, we hypothesize that EnSerp1 might be one of the key factors of host-parasite interaction. Using bioinformatic analysis of our transriptomic data we identified transcripts coding the other serine/cysteine peptidases inhibitors from different inhibitor families (e.g. Kunitz, Kazal, Bowman-Birk, α-2- macroglobulin and cystatin II inhibitors) produced by E. nipponicum and in the further research their role in the host- parasite interface should be elucidated.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112 and Masaryk University, Brno (MUNI/A/0816/2017), CEITEC project (CZ.1.05/1.1.00/02.0068), Czech Science Foundation (GBP505/12/G112), the Charles University (PRVOUK P41, UNCE 204017 and SVV 260432/2017) and European Regional Development Fund (CZ.1·05/1·1·00/02·0068)

References 1. Hahn C., Fromm B., Bachmann L. (2014) Comparative genomics of flatworms (Platyhelminthes) reveals shared genomic features of ecto- and endoparastic neodermata. Genome Biology and Evolution 6: 1105– 1117. 2. Jedličková L., Dvořáková H., Dvořák J., Kašný M., Ulrychová L., Vorel J., Žárský V., Mikeš L. (2018). Cysteine peptidases of Eudiplozoon nipponicum: a broad repertoire of structurally assorted cathepsins L in contrast to the scarcity of cathepsins B in an invasive species of haematophagous monogenean of common carp. Prasites & Vectors 11: 1–17. 3. Ilgová J., Jedličková L., Dvořáková H., Benovics M., Mikeš L., Janda L., Vorel J., Roudnický P., Potěšil D., Zdráhal Z., Gelnar M., Kašný M. (2017). A novel type I cystatin of parasite origin with atypical legumain- binding domain. Scientific Reports 7: 17526. 4. Jedličková L., Dvořáková H., Kašný M., Ilgová J., Potěšil D., Zdráhal Z., Mikeš L. (2016). Major acid endopeptidases of the blood-feeding monogenean Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae). Parasitology 143: 494–506.

Fig 1. Results of MinION nanopore sequencing. Fig 2. Mass spectrometric analyses of selected Read lengths (X axis) vs. Average read quality (Y axis). tissue dissected by laser/capture Zeiss PALM MicroBeam system. Numbers are reflecting proteins separated into 2 groups of total 731 identified.

Fig 3. Predicted 3D structure for the Eudiplozoon nipponicum stefin. The EnStef 3D structure comprises a four- strand, antiparallel beta-sheet, and a five-turn alpha-helix. (a) Motifs critical for binding to papainlike peptidases (G5G6, Q47VVAG51 and L73P74) are highlighted in light green, while the asparaginyl endopeptidase-binding domain formed by S31NS33 residues is highlighted in yellow (the numbering applies to native EnStef). (b) Surface structures included in the 3D model. (Ilgová et al. 2017).

A new genus and two new species of dactylogyrid monogeneans from gills of Neotropical catfishes (Siluriformes: Doradidae and Loricaridae) Aline A. Acosta1,2*, Tomáš Scholz2, Isabel Blasco-Costa3, Philippe Vieira Alves2,4, Reinaldo J. da Silva1 1Department of Parasitology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, São Paulo, Brazil; 2Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 3Department of Invertebrates, Natural History Museum of Geneva, Geneva, Switzerland; 4Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil *[email protected]

A new genus of dactylogyrid monogeneans (Ancyrocephalinae), Paracosmetocleithrum n. gen., is erected to accommodate P. trachydorasi n. sp. from Trachydoras paraguayensis (Siluriformes: Doradidae) in the Upper Paraná River basin, Brazil. The new genus differs from Neotropical dactylogyrids in the presence of a well-developed ornamentation in the middle portion of the ventral bar, and a sclerotised patch on the surface of the dorsal bar with an inconspicuous medial process that possesses two submedial projections arising from the tapered ends of this patch. In addition, Demidospermus rhinelepisi n. sp. is described from Rhinelepis aspera (Siluriformes: Loricariidae). The new species, which is the fifth species of the genus described from loricariids, can be differentiated from congeners by the possession of a sclerotised patch attached to the middle portion of the ventral bar, and by morphology of the accessory piece, which presents broad ends, tapering in the centre, rounded proximal end, distal end folding on both sides with folds extending to approximately ¾ of the accessory piece length. Molecular data on both new species are also provided and species composition of Demidospermus, recently revealed as polyphyletic by molecular studies including the present one, is discussed (Acosta et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Acosta A.A., Scholz T., Blasco-Costa I., Alves P.V., da Silva R.J. (2018) A new genus and two new species of dactylogyrid monogeneans from gills of Neotropical catfishes (Siluriformes: Doradidae and Loricaridae). Parasitology International 67: 4–12.

Species diversity and phylogeny of Dactylogyrus (Monogenea) parasites in Iberian Peninsula Michal Benovics1*, Andrea Šimková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

The epicontinental fauna of Iberia is strongly influenced by the geographical history of the Peninsula. Possibilities for dispersion of the organisms in this region were (and still are) limited, due to the geographical isolation caused by Pyrenees and Strait of Gibraltar, nevertheless, the latter has always been more likely bridge rather enabling migration than being a barrier for North Africa and Iberian fauna. This isolation led to the evolution of the high degree of the endemism in South Europe. Regarding the freshwater fish fauna of Iberian Peninsula, the number of families is rather low in comparison to other southern European Peninsulas, with the most species belonging to the family Cyprinidae. Representatives of this fish family serve as host for several monogenean taxa, including dactylogyrids, gyrodactylids and diplozoids. Parasites of genus Dactylogyrus exhibit narrow host specificity and their distribution is limited only on cyprinid fish. General hypothesis is that each individual cyprinid species is parasitized by its own Dactylogyrus species. Due to the narrow host specificity the evolutionary history of Dactylogyrus is intimately linked to the phylogeny and historical dispersion of cyprinids. Strong coevolutionary signal between these two taxa was revealed in the several regions across Europe. Therefore, we can expect similar pattern of Dactylogyrus – cyprinids coevolution in the Iberian Peninsula. Over years 2016 and 2017 we collected Dactylogyrus parasites from 28 host species in selected river basins across Portugal and Spain (87.5 % of all endemic cyprinids in this region). In total, 21 Dactylogyrus species were identified (14 species potentially new for science). Endemic cyprinids were parasitized by 1–5 Dactylogyrus species, where the highest parasite species richness was observed on the hosts belonging to genus Luciobarbus. Majority of potentially new species were collected from congeneric Squalius (e.g. S. aradensis, S. carolitertii, S. pyraneicus and S. torgalensis). Reconstruction of evolutionary history, based on the DNA sequences of ribosomal subunits, revealed that Iberian Dactylogyrus form 4 phylogenetic clades. While Dactylogyrus spp. of Iberian Squalius are phylogenetically closer to the Dactylogyrus spp. of Alburnus, Alburnoides and Telestes from other European regions, Dactylogyrus parasitizing Parachondrostoma and Pseudochondrostoma clustered together with the Dactylogyrus of Barbus spp. Unlike other European regions, where congeneric Barbus share same Dactylogyrus species, Iberian Barbus haasi harbour Dactylogyrus species common for local Luciobarbus (e.g. D. bocageii, D. mascomai or D. lenkoranoides). Subsequent phylogenetic reconstruction of the endemic Dactylogyrus species parasitizing Iberian fish from tribus Barbini outlined their multiple origin in this region. Additionally, we collected D. polyepidis from new host Achondrostoma arcasii. This species is morphologically similar to the D. vistulae, which is considered as the Dactylogyrus species parasitizing the widest host range. Nevertheless, D. polyepidis appears to be endemic i.e. this species is distributed only on a few cyprinid species in the Iberian Peninsula.

Acknowledgement In 2018 this study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

RNA-seq data reveal genetic variation in virulence genes between host-associated genotypes of the parasitic cnidarian Ceratonova shasta in salmonids Gema Alama-Bermejo1,2,3,4*, Eli Meyer2, Stephen D. Atkinson1, Astrid S. Holzer3, Jerri L. Bartholomew1 1Department of Microbiology, Oregon State University, Corvallis, Oregon, USA; 2Department of Integrative Biology, Oregon State University, Corvallis, Oregon, USA; 3Fish Protistology Lab, Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 4Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos Almirante Storni (CIMAS), CCT CONICET – CENPAT, San Antonio Oeste, Argentina *[email protected]

Virulence factors are essential to the success of parasite invasion, migration and spreading of infection. Myxozoans are microscopic cnidarian parasites that affect fish in culture. The myxozoan Ceratonova shasta is a complex of host- specific genotypes, infecting salmonids in the Pacific Northwest of North America. While some genotypes (I, IIC) exhibit low to moderate virulence, genotype IIR causes a profound, lethal infection in susceptible hosts. Myxozoans are unculturable, obligate endoparasites, and thus deep sequencing has been fundamentally challenging due to host contamination and their fast molecular evolution, which yields strongly divergent sequences. We developed new pipelines to obtain host-free comprehensive myxozoan RNA-seq data from infections in three different salmonid fishes i.e. rainbow trout Oncorhynchus mykiss, coho salmon Oncorhynchus kisutch and Chinook salmon Oncorhynchus tschawytscha in which the same parasite species shows different virulence. We focused on two genetic targets strongly-related to virulence: cell migration genes, and proteolytic enzymes and their inhibitors, and we identified genetic variations of these factors between C. shasta genotypes. We developed a two- step bioinformatics pipeline to generate a reference transcriptome of the most virulent genotype (IIR) for discovery of candidate virulence genes. This study is the first to compare the genetic variation among myxozoan genotypes by searching for single nucleotide polymorphisms (SNPs) in Next Generation Sequencing data. We confirmed the limited resolution of established Internal Transcribed Spacer markers of C. shasta genotypes by transcriptome-wide genetic distances analyses, which showed that genotype II represents a mix of two types with different host association (IIC for coho salmon and IIR for rainbow trout), with relatively large genetic distance. Analyses of SNPs in gene groups associated with virulence showed that cell migration genes are most variable between parasite genotypes I and II, while proteases are most variable between genotype IIR and all other genotypes tested. We demonstrate that variations in protease genes are strongly associated with virulence in C. shasta, and we suggest that this group of enzymes is a promising candidate for targeted future interventions against myxozoans in aquaculture.

Future plans A manuscript has been submitted to a scientific journal and is under review. Molecular and quantitative gene expression analyses were performed on evaluation of motility genes and proteases + inhibitors as virulence factors for genotypes of C. shasta. Two further publications are under preparation.

Acknowledgement This study was supported by Czech Science Foundation (P505/12/G112 and 14–28784P); Consellería de Educación, Investigación, Cultura y Deporte, Valencia, Spain (APOSTD/2013/087) and The United States Department of the Interior, Bureau of Reclamation (R15PG00065).

Towards a robust systematic baseline of Neotropical fish tapeworms (Cestoda: Proteocephalidae): amended diagnoses of two genera from the redtail catfish, Phractocephalus hemioliopterus Philippe V. Alves1,3, Alain de Chambrier2, José L. Luque1, Tomáš Scholz3* 1Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil; 2Department of Invertebrates, Natural History Museum of Geneva, Geneva, Switzerland; 3Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

Species of two monotypic genera, Ephedrocephalus Diesing, 1850 and Zygobothrium Diesing, 1850 (Cestoda: Proteocephalidae), parasites of one of the most basal members of the catfish family Pimelodidae, the redtail catfish Phractocephalus hemioliopterus, in the Neotropical Region, are redescribed based on the evaluation of type specimens and newly collected material. Generic diagnoses are amended to provide a robust baseline for the future re-arrangement of the classification of proteocephalid cestodes. Ephedrocephalus is typified by the medullary position of the ovary and uterus, the cortical distribution of vitelline follicles (dispersed throughout almost the entire ventral cortex) and the testes in one dorsal field. Zygobothrium is primarily characterized by its possession of a robust scolex bearing four uniloculate suckers with two openings each and by the tetralobed velum (laciniations) on every proglottid (two on the ventral and two on the dorsal side). The redtail catfish is the definitive host of as many as seven species of proteocephalid cestodes, which, however do not represent a monophyletic group. Some species, including Z. megacephalum, are among the earliest diverged parasites of Neotropical catfishes, being closely related to African and North American proteocephalids from catfishes, whereas others such as E. microcephalus belong to more recently diverged taxa with uncertain interrelations. Unlike most proteocephalids of the redtail catfish which almost always infect the anterior parts of the host intestine, E. microcephalus and Z. megacephalum occur exclusively in its posterior third. A key to the identification of the proteocephalid species parasitizing P. hemioliopterus is also presented (Alves et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Alves P.V., de Chambrier A., Luque J.L., Scholz T. (2018) Towards a robust systematic baseline of Neotropical fish tapeworms (Cestoda: Proteocephalidae): amended diagnoses of two genera from the redtail catfish, Phractocephalus hemioliopterus. Zootaxa 4370(4): 363–380.

Experimental evaluation of behavioural changes in gilt-head seabream infected with brain-encysted metacercariae of Cardiocephaloides longicollis (Trematoda, Strigeidae) Gabrielle S. van Beest1,2, Francisco E. Montero1, Juan A. Raga1, Ana Born-Torrijos1,2* 1Cavanilles Institute for Biodiversity and Evolutionary Biology, Science Park, University of Valencia, Valencia, Spain; 2Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

Trophically transmitted parasites may increase their transmission efficiency by altering the behaviour of infected hosts, thus increasing their susceptibility to predation by the next host. The strigeid trematode Cardiocephaloides longicollis (Rudolphi, 1819) Dubois, 1982 parasitizes 31 fish species, including the gilt-head seabream (Sparus aurata L.), one of the most important fish in Mediterranean aquaculture. Actually, this parasite has recently been found in gilt-head seabreams from aquaculture facilities, with a prevalence of infection up to 54 % [1]. The cercariae penetrate the skin and migrate into the brain, where they encyst as metacercariae, and the definitive hosts, seabirds, become infected by consumption of infected fish. It is commonly believed that the parasite larvae can cause significant alterations in fish behaviour, thus increasing their transmission to the definitive host, as evidenced in other brain- infecting trematodes (e.g. Euhaplorchis californiensis [2]). However, the behavioral pathology suggested to be provoked by C. longicollis has never been experimentally studied. In this study, an experiment to detect differences in the behaviour of infected vs. uninfected fish was performed. First, 14 fish were experimentally infected with 180 cercariae of C. longicollis, emerged from their first intermediate host, the snail Nassarius reticulatus. Preliminary assays showed an infection success around 50 %, so that a high number of metacercariae should be accumulated in the fish brain. Furthermore, behaviour experiments were run 6 months post-infection to ensure that metacercariae were infective. Fish were placed in a plexiglass column (200 cm height, 30 cm diameter) where an effective light-dark gradient was generated. The water column was vertically divided into 20 cm sections and the position of each fish at 1-min intervals for 30 minutes every 2 hours during 3 days was recorded in three assays, i.e. control (i.e. uninfected), infected and mixed fish group. Preliminary results show significant differences in the distribution of control and infected fish along the sections in the tube, in both separated and mixed groups, suggesting a different use of the space, and thus behaviour. This may indicate that encysted metacercariae might provoke this behavioral alteration in infected fish within the tube associated to a neuronal disorder. Despite of the fact that most metacercariae infect the optical lobes, this behaviour might not be a consequence of a decrease of light perception, as infected fish occupied generally deeper and so darker positions. However, other aspects of fish vision to be important in the nervous control of behaviour or the host´s antipredator responses, such as visual acuity, could be affected, which has to be further studied.

Future plans Finalisation of the statistical analyses of the data produced during this research, as well as realization of further experiments studying antipredator responses of parasitized fish. Thereafter, preparation and finalisation of manuscripts for publication.

Acknowledgement This study was supported by projects MSM200961706 (Czech Academy of Sciences), AGL2015-68405-R (MINECO/FEDER, UE), Prometeo/2015/018 and Revidpaqua ISIC/2012/003 (Valencian Regional Government), as well as by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

References 1. Born-Torrijos A., Poulin R., Perez-del-Olmo A., Culurgioni J., Raga J.A., Holzer A.S. (2016) An optimised multi- host trematode life cycle: fishery discards enhance trophic parasite transmission to scavenging birds. International Journal for Parasitology 46: 745–753. 2. Lafferty K.D, Morris A.K. (1996) Altered behavior of parasitized killifish increases susceptibility to predation by bird final hosts. Ecology 77: 1390–1397.

Use of in vivo fluorescent dyes for tracking the penetration of Cardiocephaloides longicollis (Digenea: Strigeidae) into the gilthead sea bream Sparus aurata Gabrielle S. van Beest 1,2*, Francisco E. Montero1, Mar Villar-Torres1, Juan A. Raga1, Ana Born-Torrijos1,2 1Cavanilles Institute for Biodiversity and Evolutionary Biology, Science Park, University of Valencia, Valencia, Spain; 2Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

Digenean trematodes are a recurrent model system for having a better understanding of the evolution of animal life history traits and behaviors. Studying their host-parasite interactions can be a big challenge due to their complex life cycle involving several hosts and free larval stages. The trematode parasite Cardiocephaloides longicollis (Rudolphi, 1819) (Digenea: Strigeidae) is widespread along European coastlines. It parasitizes the gilthead sea bream (Sparus aurata L.), one of the most important marine fish in the Mediterranean aquaculture, with up to 54 % prevalence [1]. Cardiocephaloides longicollis has a three-host life cycle, where the cercariae, after being released by snail, first intermediate host, penetrate the skin and migrate into the fish brain, where they encyst as metacercariae. The cysts could cause significant alterations in fish behaviour facilitating their transmission to the definitive host [2]. For the study of the entry portals of cercariae into the fish, in vivo fluorescent dyes were used: (1) 5(6)- carboxyfluorescein N-hydroxysuccinimidyl ester (CFSE), which stains the acetabular gland of the cercariae, and (2) Hoechst 33342 (NucBlue, NB), which specifically stains DNA (i.e. the nuclei of cells). Both were tested in a short and long term experimental assay. For the short term assay, three ascending concentrations of each dye were applied to detect the effects on cercarial survival within 24 hour post-labelling (hpl) and 5 hpl, and on cercarial activity within 5 hpl (the time during which cercariae should be highly infective). For the long term assay, CFSE or NB-labelled cercariae with intermediate concentrations were used to infect fish and record their infection success (encysted metacercariae in the brain). Intermediate doses of CFSE did not show any short term effect on cercarial survival and very low effect on their activity. However, NB seemed to be more adequate for long term assays as it did not affect cercarial infection and encystment in the brain. Therefore, CFSE was used to determine the penetration points of C. longicollis into the fish host, detecting aggregations of cercariae especially on the head, the eyes and gills region, as well as on the dorsal fin and the lower side. Thus, areas nearby the brain (target organ) or well connected to the brain through neuronal canals seem to be the most attractive to C. longicollis cercariae. In conclusion, the in vivo fluorescent dye CFSE is the most adequate labelling treatment in short term experimental assays, such as those studying the penetration strategy of cercariae. This helped to recognize the entry portals of C. longicollis into the gilthead sea bream, located strategically on areas close to the brain or well connected to it through the neuronal canals.

Future plans Experimental assays are currently being performed to increase our understanding of the life cycle of C. longicollis. One of our aims is to determine the migration route of C. longicollis cercariae within the fish towards the brain, by histological samples of sequential artificial infections to track cercariae within the fish body during the infection process.

Acknowledgement This study was supported by projects MSM200961706 (Czech Academy of Sciences), AGL2015-68405-R (MINECO/FEDER, UE), Prometeo/2015/018 and Revidpaqua ISIC/2012/003 (Valencian Regional Government), as well as by the Czech Science Foundation (project No. P505/12/G112).

References 1. Born-Torrijos A., Poulin R., Pérez-del-Olmo A., Culurgioni J., Raga J.A., Holzer A.S. (2016) An optimised multi- host trematode life cycle: fishery discards enhance trophic parasite transmission to scavenging birds. International Journal for Parasitology 46: 745–753. 2. Osset E.A., Fernández M., Raga J.A., Kostadinova A. (2005) Mediterranean Diplodus annularis (Teleostei: Sparidae) and its brain parasite: Unforeseen outcome. Parasitology International 54: 201–206.

From taxonomic deflation to new cryptic species: Hidden diversity in a widespread African squeaker catfish Dagmar Jirsová1,2*, Jan Štefka1,2, Radim Blažek3,4, John O. Malala5, David E. Lotuliakou5, Zuheir N. Mahmoud6, Miloslav Jirků2 1Faculty of Science, University of South Bohemia, České Budějovice, Czech Republic; 2Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 3Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic; 4Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 5Kenya Marine and Fisheries Research Institute, Lake Turkana Station, Lodvar, Kenya; 6Department of Zoology, Faculty of Science, University of Khartoum, Khartoum, Sudan *[email protected]

Cryptic genetic diversity and erroneous morphological species determination represent frequent problems in biodiversity research. Here, examination of 138 specimens of Synodontis (Mochokidae, Siluriformes) from the Nile River and Lake Turkana revealed presence of both S. schall-like and S. frontosus-like morphotypes, with phenotypic gradient between them. Phylogenetic and haplotype analyses based on two mitochondrial and one nuclear marker including 131 coxI (565bp), 96 cytb (973bp) and 23 RAG2 (896bp) sequences from the Nile-Turkana population, plus additional GenBank sequences of Synodontis spp., suggest that both morphotypes and intermediate forms are conspecific. The results imply probable synonymy of S. frontosus with S. schall. Strong biogeographical signal has been revealed among supposedly widely distributed, homogenous and conspecific S. schall-like catfish of the Nilo- Sudanian ichthyological province using the mitochondrial dataset. Synodontis schall sensu stricto, as defined by type locality in the Nile, is apparently restricted to eastern part of the Nilo-Sudanian ichthyological province (e.g. Nile, Turkana, Chad), while S. schall sensu lato from western parts (e.g. Senegambia, Niger, Chad) most probably represents a cryptic taxon unrecognized thus far due to the absence of tenable morphological differences. There is a geographic overlap between the eastern “sensu stricto” and western “sensu lato” clades in the Chad basin.

Future plans Manuscript has been currently resubmitted after major revision and waits editorial decision.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

Spring viremia of cyprinid species and their hybrids Kristina Civáňová1*, Ondřej Pěnčík1, Andrea Šimková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

Viral diseases in aquacultures are serious problem mainly due to lack of treatment options. Spring viremia of carp (SVC) is caused by Rhabdovirus carpio, a bullet-shaped RNA virus. The disease was initially diagnosed in Yugoslavia; subsequently, it has been reported in many regions of the world, mainly in Europe, Middle East and also in America. The following hosts were reported for SVCV: common carp (or koi carp) (Cyprinus carpio), grass carp (Ctenopharyngodon idella), bighead carp (Aristichthys nobilis), silver carp (Hypophthalmichthys molitrix), and also Carassius species (C. carassius, C. auratus gibelio, C. auratus auratus). Mortality caused by SVCV has reached 70 % in yearling carp from European populations. Adult fish can also be affected but to a lesser degree. The disease may turn into a chronic stage. In our work, we focused on SVCV mechanism, development and progress. In this study, we examined the level of expression of SVCV in C. carpio, C. auratus gibelio and their F1 hybrids. Several hypotheses have been predicted for parasite load in parental species and their F1 hybrids (dominance, resistance, susceptibility or additive scenario). The main aim was to investigate the SVCV infection progress in three above mentioned groups of fish. During introductory experiments, 3 specimens of each fish group were analysed. RNA was isolated (PAXgene Blood RNA Kit, Qiagen) from blood samples collected in exactly defined times before (0 hpi) and after infection (6/12/24/48 hours post infection; 3/5/7/10 days post infection). In total, 81 RNA samples were used as a template for one-step real-time quantitative PCR (qRT-PCR) to detect virus presence in the sample and to determine viral copy number. For this purpose, commercial SVCV genesig advanced kit (Primerdesign, Ltd; Qiagen) according to recommendations of manufacturer and CFX96 Real-Time PCR Detection System (Bio-Rad) were used. To precise calculation of viral copy number in each sample, the obtained cq values of each sample were compared with the cq of constructed standard curve and the measured RNA concentration of each isolate (NanoDrop 8000, Thermo Scientific) was taken into account. Evaluation of results was performed using CFX Manager software (Bio-Rad). Our preliminary findings about viral expression within host groups correspond with previous studies in this field [1,2]. In C. gibelio, at 12 hpi, SVCV gene for L2 inner capsid protein VP2 was detected in all fish. Viral copies rapidly increased in all samples at 24-48 hpi to the maximum. In C. carpio, slower progress of an infection was observed and SVCV target was detected later (24 hpi); viral copies increased to maximum at 3–5 dpi in this group. In the hybrid group, the infection progress was very heterogenic between individuals and could not be characterized uniformly. However, results in F1 hybrid generation indicate higher impact of maternal genes on immunity response of particular hybrid individual. After reaching the maximum, viral copies were gradually reduced; i.e.7 dpi, no or minimum viral copies were detected in all samples of all groups. More individuals and different hybrid combinations are necessary for the following studies about spring viremia of carp virus infection and modulation of this infection in hybrid lines. The regulation of SVCV infection may be potentially useful in process of eradication of introduced species, immune response research using breed lines, or in development of fish antivirotics (vaccines or antiviral treatment).

Future plans Continue in qRT-PCR analyses for SVCV copy number detection using larger sample of individuals within groups of parental species and hybrids and also using more hybrid combinations. The obtained results will be used in planned subsequent analysis of expression of immune genes in fish involved in immune response to find out the correlation between immune genes and viral copy number.

Acknowledgement This study was supported by European Centre of Ichthyoparasitology, Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

References 1. Wei X., Li X. Z., Zheng X., Jia P., Wang J., Yang X., Yu L., Shi X., Tong G., Liu H. (2016) Toll- like receptors and interferon associated imine factors responses to spring viraemia of carp virus infection in common carp (Cyprinus carpio). Fish and Shellfish Immunology 55: 568–576. 2. Ahne W., Bjorklund H.V., Essbauer S., Fijan N., Kurath G., Winton J.R. (2002) Spring viraemia of carp (SVC), Diseases of Aquatic Organisms 52(3): 261–272.

Diplozoid species of endemic cyprinids from Mediterranean area Kateřina Čermáková1*, Božena Koubková1, Imane Rahmouni2, Kristína Civáňová1, Andrea Šimková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic 2Laboratory of Zoology and General Biology, Faculty of Sciences, Mohammed V University in Rabat, Morocco *[email protected]

The Mediterranean area is known for the presence of many endemic fish species. Cyprinids represent the most diverse and most abundant freshwater fish groups in this area. However, the fauna of their parasites is poorly known. In this study, a total of 135 endemic cyprinid species collected during 2014–2018 were investigated for the presence of diplozoid monogeneans covering almost whole range of Euro-Mediterranean area and a part of African Mediterranean area (Morocco). Diplozoid species were determined based on the morphology of the attachment apparatus and identification was confirmed by the molecular analysis using partial 18S rDNA, 28S rDNA, ITS1 and ITS2 regions. Morphological determination of over 1100 specimens of diplozoids together with phylogenetic analyses revealed five species of genus Paradiplozoon. The most abundant diplozoid species reported on endemic cyprinids of Mediterranean was Paradiplozoon homoion, common parasite of several species of cyprinids in Central Europe. This study provides 17 new host records of P. homoion – i.e. Telestes montenigrinus from Albania; Chondrostoma knerii and Scardinius plotizza from Croatia; Alburnoides oeconomui, A. thessalicus, Barbus balcanicus, B. sperchiensis, Pelasgus marathonicus, Rutilus sperchios, R. ylikiensis, Telestes beoticus and T. alfiensis from Greece; and Achondrostoma arcasii, Barbus haasi, Luciobarbus graellsii. Parachondrostoma miegii and Squalius laietanus from Spain. Furthermore, we determined Paradiplozoon megan, known as a parasite of genera Squalius and Leuciscus in Central Europe, on five host species – i.e. Squalius zrmanjae and S. squalus from Croatia and S. lucumonis, S. squalus, Protochondrostoma genei and Rutilus rubilio from Italy. In addition, we found three species of Paradiplozoon representing potentially new species for science. Paradiplozoon sp. 1 parasitizing Scardinius acarnanicus and Tropidophoxinellus hellenicus from Greece is phylogenetically and morphologically closely related to P. megan. Paradiplozoon sp. 2 was recorded on 9 species of cyprinids in Iberian Peninsula, i.e. Iberochondrostoma lusitanicum and Luciobarbus bocagei from Portugal; and Luciobarbus bocagei, L. guiaronis, Pseudochondrostoma polylepis, Squalius carolitertii, S. pyrenaicus, S. valentinus, Parachondrostoma arrigonis and P. turiense from Spain. This species is phylogenetically closely related to P. pavlovskii. Paradiplozoon sp. 3 was reported only on Moroccan Luciobarbus lepineyi and is phylogenetically closely related to P. bingolensis from Garra rufa from Turkey.

Future plans Finalization of publication concerning the distribution and phylogeny of Mediterranean diplozoids (including also the description of new species).

Acknowledgement This study was financially supported from 2018 by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project no. P505/12/G112. We thank Radek Šanda (National Museum, Prague) and Jasna Vukić (Charles University, Prague) for the fish sampling.

Comparative phylogenetic analysis of Apicomplexa based on 18s, 28s and contig 18s+28s rDNA Andrei Diakin1*, Timur G. Simdyanov2, Andrea Valigurová1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2Department of Invertebrate Zoology, Faculty of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia. *[email protected]

Apicomplexans form a large and diverse group of unicellular parasites inhabiting invertebrates and vertebrates. Some of them are responsible for important human and animal diseases (e.g. Plasmodium spp., Toxoplasma gondii, and Cryptosporidium spp.), therefore are intensively studied in various aspects of biology and medicine. However, basal groups (e.g. gregarines, agamococcidia, blastogregarines, and protococcidia) that are restricted to invertebrate hosts remain poorly understood. There are a lot of investigations dedicated to intraspecies (relationship between different strains) and intragenus phylogenetic relationships of aforementioned pathogens. Less investigations concern molecular phylogeny of other apicomplexan groups, and even less are dedicated to phylogeny of Apicomplexa as whole. We performed comparative analysis of several phylogenetic trees: 1. 18S that include large most available taxa; 2. 18s tree with reduced number of taxa (the taxa were selected based on 28S set of sequences; 3. 28S phylogenetic tree; 4. 18S+28S phylogenetic trees. Large 18S tree fits to a modern view on presence of the main groups of Apicomplexa, however, as expected, ordering and supports are variously depending on selected analysis. Reduced 18S tree shows another picture: despite the presence of main clades, some species jumped to the other clades; other point is that the ordering does not fit to a large 18S tree. 28S and 18S+28S trees show almost the same results to each other and to 18S tree with a large number of taxa, with flipping of some clades, however. Other consequences is the increasing of clades supports. We can conclude that for better understanding of evolutionary pathways we need to process more data. In addition the use of multigene analysis would improve the results of analysis and data interpretation.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112. Authors acknowledge Gita G. Paskerova (Dep. Of Invertebrte Zoology, Faculty of Biology, Saint-Petersburg State University) for her help in field sampling. Vladimir V. Aleoshin (Belozersky Institute for Physico-Chemical Biology, M.V. Lomonosov Moscow State University).

Crystalline inclusions in “small” amphizoic amoebae Iva Dyková1*, Tomáš Bílý2, Tomáš Tyml1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2Laboratory of Electron Microscopy, Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budĕjovice, Czech Republic *[email protected]

The first reports on the presence of crystal-like structures in the cytoplasm of amoebae are dated back to the 18th century when pioneering studies of famous microscopists (Joseph Leidy, Eugène Penard, Johannes Frenzel and others) were conducted on living cells of Amoeba, Chaos, and Pelomyxa spp. For understandable reasons, these large/giant free-living amoebae for a long time remained objects of interest of light microscopists [1] The shape, size and number of crystals were used to characterize individual amoeba species or even establish new ones (e.g., Amoeba crystalligera Gruber, 1885; Mayorella crystallus Schaeffer, 1926; Trichamoeba osseosaccus Schaeffer, 1926). The size (up to 5 µm in length in bipyramidal crystals) and quantity of crystals obtainable from giant amoebae facilitated early attempts to supplement light microscopical observations of crystals with x-ray crystallography and analysis of physicochemical properties [6,3,4]. The extension of the use of transmission electron microscopy turned attention to crystalline inclusions of many types of cells [2] including small amoebae, which, however, so far, have remained on the edge of interest. Based on our studies of diversity of amoebae, an integral part of which is also a study of their ultrastructure, incidental findings of crystalline structures are demonstrated in three strains of amphizoic amoebae: Acanthamoeba strain 4337 isolated from the brain of chub (A), Neovahlkampfia strain UD12 isolated from the gill tissue of rainbow trout (B) and ACN1 strain of Vannella isolated from the gills of turbot (C–F). In two of these strains (ACN1 and 4337), crystalline structures were found in the cytoplasm, whereas in UD12, similar structures were localized in the nucleus (B). Crystalline inclusions demonstrated in the electron micrographs have in common a highly ordered pattern of closely spaced parallel dense lines made up of globular units (seen at high magnifications of JEOL JEM 1010). The orientation of sections with respect to plane of the crystal lattice determined the limited possibilities of comparison of our findings with crystal-like structures observed in ultrathin sections by other authors. Curiously enough, a remarkable similarity was observed between crystalline structures in trophozoites of ACN1 strain and those found in hepatic cells of the slender salamander Batrachoseps attenuatus [2]. The nature and biological significance of crystalline structures is not clear in either our amoebae or the hepatic cells. To obtain more data, our studies of ACN1 strain crystals (those most frequent in ultrathin sections of our material) continue at an advanced methodical level using electron tomography processed by IMOD software [5].

Acknowledgement This research was supported by the Czech Science Foundation (grant 505/12/G112) and the Ministry of Education, Youth and Sports (grant LM2015062 Czech-BioImaging).

References 1. Bovee E.C., Jahn T.L. (1973) Taxonomy and Phylogeny. Chapter 2. In: Jeon K.W (Ed). The Biology of Amoeba. Academic Press New York and London, pp. 628. 2. Fawcet D.W. (1966) The Cell. An Atlas of Fine Structure. W.B. Saunders comp. Philadelphia and London, pp. 448. 3. Griffin J.L. (1960) The isolation, characterization and identification of the crystalline inclusions of the large free-living amoebae. Journal of Biophysics and Biochemical Cytology 7: 227–234. 4. Grundbaum B.W., Møller K.M., Thomas R.S. 1959. Cytoplasmic crystals of the amoebae: Amoeba proteus and Chaos chaos. Experimental Cell Research 18: 389–391. 5. Kremer J.R., Mastronarde D.N., McIntosh J.R. (1966) Computer visualization of three-dimensional image data using IMOD. Journal of Structural Biology 116: 71–76. 6. Luce R.H., Pohl A.W. (1935) Nature of crystals found in amoeba. Science 82: 595–596.

Fig 1., A. An overview of crystalline structure localized in the cytoplasm of Acanthamoeba (strain 4337 isolated from brain of chub); B. The nucleus of Neovahlkampfia trophozoite (strain UD12 isolated from raibow trout gill tissue). Crystalline structure marked with arrowheads; C, D. Electron dense crystal-like structures frequently observed in the cytoplasm of Vannella (ACN1 strain isolated from gills of turbot); E, F. Crystalline structures observed under high magnifications reveal a highly ordered pattern of closely spaced, parallel dense lines representng one-plane view of crystal lattices.

Does classification of Schilbetrema and Schilbetrematoides (Monogenea: Dactylogyridae) reflect their molecular phylogeny? Kateřina Francová1*, Mária Seifertová1, Eva Řehulková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

Schilbetrema and Schilbetrematoides species (Monogenea, Dactylogyridae) are gill parasites of schilbeid catfishes (Siluriformes, Schilbeidae) with distribution restricted to African region. To date, fourteen Schilbetrema and two Schilbetrematoides species have been recorded to infect schilbeids (Schilbe intermedius, S. mandibularis, S. mystus, Parailia pellucida). Species of Schilbetrema Paperna and Thurston, 1968 are characterized, in part, by having ventral anchor with a prominent superficial knob on its base. The other characters defining the members of this genus include presence of ventral and/or dorsal haptoral bars with lateral, subterminal, and/or submedial anterior projections. Species of Schilbetrematoides Kritsky and Kulo, 1992 are unusual worms with apparent close relationship to Schilbetrema (based, among others, on identical basic features of reproductive system) but also having a reduced ventral anchor-bar complex. During our survey in Kenya (Lake Turkana), Sudan (River Blue Nile and River White Nile) and Cameroon (River Boumba), we examined three host species (Schilbe intermedius, S. mystus and S. uranoscopus) and collected a majority (eight) of the previously described Schilbetrema species and one previously described Schilbetrematoides species. Further, we recorded three species new to science (two Schilbetrema spp. and one Schilbetrematoides sp.). This study thus presents an extensive work on monogeneans of schilbeids, including their morphological characterization, assessment of genetic divergence and phylogenetic relationship between members of Schilbetrema and Schilbetrematoides, based on the partial 18S, entire ITS1, and partial 28S rDNA sequences. Our phylogenetic analysis showed that monogenean species studied were distributed in two separate clades. The A clade was divided into three well-supported sister branches: two Schilbetrema branches (each including three species), and one branch including all (two) Schilbetrematoides species. The second clade (B) comprised the rest of Schilbetrema species (four species). The phylogenetic position of Schilbetrema species well corresponded to particular morphological type, based mainly on morphology of ventral and dorsal bar. Considering the morphological and molecular variety within Schilbetrema, and also phylogenetic position of Schilbetrematoides spp. in relation to Schilbetrema spp., the current classification of monogeneans infecting schilbeids is questionable.

Acknowledgement This research was supported by the Czech Science Foundation, project no. P505/12/G112 (ECIP).

Microsatellite markers – valuable tools for population genetic study of Dactylogyrus vistulae Lenka Gettová1*, Andrea Šimková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

Majority of the population genetic studies of monogenean parasites are based on the rRNA subunits and ITS region. On the other hand, population studies on Monogenea based on nuclear markers, such as microsatellites, are scarce. Dactylogyrus vistulae Prost, 1957 represents a generalist monogenean parasite with a high number of fish host species and widespread Eurasian distribution. Therefore, D. vistulae represent a suitable model species for microsatellite markers development and their application in population genetic studies. In our study, we aimed to optimize five multiplex sets of 25 de novo isolated polymorphic microsatellite markers and test their suitability for study of genetic structure and variability of D. vistulae populations. D. vistulae (N = 215) infecting 31 fish species from six European countries (Czech Republic, Italy, Croatia, Bosnia and Herzegovina, Albania, and Greece) were sampled. Various programs (i.e. GENETIX, GenAlEx, Fstat) were applied in order to study genetic structure and variability of D. vistulae populations using microsatellite markers. Factorial correspondence analysis (FCA) calculated in GENETIX divided D. vistulae individuals into three clusters separating D. vistulae populations from Bosnia and Herzegovina and majority of individuals from Croatia from main cluster which comprised remaining individuals. However, clustering based on country of collection was still visible inside this cluster. Moreover, D. vistulae individuals which parasitized the same host species tended to cluster together. Differences in genetic variability of D. vistulae parasitizing different host species were found, revealing higher genetic variability of D. vistulae infecting widespread fish species than endemic ones. Our study revealed that microsatellites represent valuable tools for population genetic study of D. vistulae. Since flanking regions of microsatellites are often conserved in congeneric taxa, de novo isolated microsatellites for D. vistulae might be also used for study of genetic variability and structure of other monogenean species. Study involving cross-species amplification is further needed to confirm this hypothesis.

Future plans Cross-species amplification. Preparation of manuscript.

Acknowledgement This study was financially supported by ECIP - Centre of excellence, GACR No P505/12/G112. We thank Michal Benovics and Kristýna Koukalová for providing D. vistulae samples and laboratory help, respectively.

Philometrid nematodes of marine fishes from the Gulf of Hammamet, Tunisia David G. Solís1,2*, Nessrine Ghanmi3 1El Colegio de la Frontera Sur (ECOSUR), unidad Chetumal. Av. Centenario km. 5.5, Chetumal, Mexico; 2Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 3Unité de Recherche Bio-Écologie Animale et Systématique Évolutive, Faculté des Sciences, Université Tunis El Manar, Tunis, Tunisia *[email protected]

Nematodes belonging to the family Philometridae represent a diverse group of parasites infecting freshwater brackishwater and marine teleost fishes around the world. They have a very particular morphology and biology, and exhibit a high host and site specificity and a marked sexual dimorphism, where females are considerably larger than their conspecific males (Moravec and de Buron, 2013). Additionally, most philometrids have been described based solely on the conspicuous females, whereas males are unknown in several valid species. The Mediterranean Sea is one of the regions where these nematodes have been reported in various fish families (e.g. Serranidae, Uranoscopidae, Mullidae). Recent collections of nematodes from marine fishes resulted in the discovery of three undescribed species, as well as unknown males of Philometra serranellicabrillae. The new species, P. draco in Trachinus draco, P. radiata in Trachinus radiatus and P. barbata in Mullus barbatus, differ from their congeners in the different structures on the gubernaculum (e.g. smooth field between the two longitudinal lamellated parts, dorsal protuberance), as well as in the body, spicules and gubernaculum lengths (Ghanmi et al., 2018a, b, c). These findings increase the number of philometrids reported from the Mediterranean Sea to 15 species. Moreover, the male of P. serranellicabrillae is reported for the first time from the type host, but in a different locality. The morphology of its posterior end is different from those of males of other species in the shape of the caudal reniform mounds, which are clearly divided in dorsal and ventral lobes. Apparently, there is an important philometrid fauna in marine fish from the Mediterranean region, which could be better described in the near future when species of other fish families are examined.

Future plans Compilation of data on nematodes of marine fishes from the American continent for a monograph.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112 and by the Ministry of Higher Education and Scientific Research of Tunisia and Unité de Recherche Bio-Écologie Animale et Systématique Évolutive, Faculté des Sciences, Université Tunis El Manar, Tunisia.

References 1. Moravec F., de Buron I. (2013) A synthesis of our current knowledge of philometrid nematodes, a group of increasingly important fish parasites. Folia Parasitologica 60: 81–101. 2. Ghanmi N., González-Solís D., Gargouri L. (2018a) Two new gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) from Trachinus spp. (Osteichthyes: Trachinidae) in the Gulf of Hammamet, Tunisia. Systematic Parasitology 95: 223–234. 3. Ghanmi N., González-Solís D., Gargouri L. (2018b) Philometra barbata n. sp. (Nematoda: Philometridae) from the red mullet Mullus barbatus (Perciformes, Mullidae) off Tunisia. Acta Parasitologica 63: 766–771. 4. Ghanmi N., González-Solís D., Gargouri L. (2018c) First description of the male of Philometra serranellicabrillae Janiszewska, 1949 (Nematoda: Philometridae), a gonad-infecting parasite of the marine fish Serranus cabrilla (Serranidae) off Tunisia (in preparation).

Fig 1. Philometra draco n. sp. ex Trachinus draco, scanning electron micrographs of male. A, B. Cephalic end, apical and sublateral view, respectively; C. Posterior end, apical view (arrows indicate phasmids and arrowheads indicate papillae); D. Posterior end showing spicules and gubernaculum, lateral view; E. Spicules and gubernaculum, sublateral view; F. Same, higher magnification, subapical view. Abbreviations: a, amphid; b, submedian pair of cephalic papillae of outer circle; c, cephalic papilla of inner circle; g, gubernaculum; s, spicule (tomado de Ghanmi et al., 2018a).

The joint evolution of the Myxozoa and their alternate hosts: a history of host acquisitions and massive diversification events Astrid S. Holzer1*, Pavla Bartošová-Sojková1, Ana Born-Torrijos1, Alena Lövy1, Ivan Fiala1 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice; Czech Republic *[email protected]

The relationships between parasites and their hosts are intimate, dynamic and complex; the evolution of one is inevitably linked to the other. Despite multiple origins of parasitism in the Cnidaria, only parasites belonging to the Myxozoa are characterized by a complex life cycle, alternating between fish and invertebrate hosts, as well as by high species diversity. This inspired us to examine the history of adaptive radiations in myxozoans and their hosts by determining the degree of congruence between their phylogenies and by timing the emergence of myxozoan lineages in relation to their hosts. Recent genomic analyses suggested a common origin of Polypodium hydriforme, a cnidarian parasite of acipenseriform fishes, and the Myxozoa, and proposed fish as original hosts for both sister lineages. We demonstrate that the Myxozoa emerged long before fish populated Earth and that phylogenetic congruence with their invertebrate hosts is evident down to the most basal branches of the tree, indicating bryozoans and annelids as original hosts and challenging previous evolutionary hypotheses. We provide evidence that, following invertebrate invasion, fish hosts were acquired multiple times, leading to parallel cospeciation patterns in all major phylogenetic lineages. We identify the acquisition of vertebrate hosts that facilitate alternative transmission and dispersion strategies as reason for the distinct success of the Myxozoa, and identify massive host specification-linked parasite diversification events [1]. A follow-on study obtained numerous DNA sequences of the most basal hard-shelled myxozoans (Sphaerospora sensu strico) and studied cophylogenetic events in this poorly characterized group [2]. The presence of two separate phylogenetic lineages in Sphaerospora s. str. likely indicates independent historical host entries, and the remarkable overlap of the larger clade with vertebrate phylogeny confirms coevolutionary findings from other clades. Sequencing of sphaerosporids from cartilaginous fish, or other evolutionary older vertebrate groups could substantially support this idea and shine a light on the early evolution of myxozoans in fish. Overall, the results of our studies transform our understanding of the origins and evolution of parasitism in the most basal metazoan parasites on Earth.

Future plans Molecular dating analyses of myxozoans demonstrated that first became parasitic in archiannelids. Basal to the known annelid hosts of myxozoans two groups can be found, Haplodrili (= Archiannelida, five families) and Sipuncula. We have collected a large number of specimens from these taxa and are sequencing their DNA to determine if myxozoans of basal lineages can be found in these organisms. We have also obtained 17 new sequences of myxozoans from evolutionary old fishes (sharks and rays), including 3 new lineages and we analyse these sequences in the light of life cycle expansion and acquisition of a second host.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112, as well as the Horizon2020 Research and Innovation Action project ParaFishControl (reference #634429). The authors thank Prof. Christopher Secombes and Dr. Jason Holland (University of Aberdeen) as well as Prof. Jerri Bartholomew and Dr. Stephen Atkinson (Oregon State University) for providing genome/transcriptome sequence data of Tetracapsuloides bryosalmonae and Ceratonova shasta.

References 1. Holzer A.S., Bartošová-Sojková P., Born-Torrijos A., Lövy A., Hartigan A., Fiala I. (2018) The joint evolution of the Myxozoa and their alternate hosts: A cnidarian recipe for success and vast biodiversity. Molecular Ecology 27: 1651–1666. 2. Patra S., Bartošová-Sojková P., Pecková H., Fiala I., Eszterbauer E., Holzer A.S. (2018) Biodiversity and host- parasite cophylogeny of Sphaerospora (sensu stricto) (Cnidaria: Myxozoa). Parasites & Vectors 11: 347.

Recombinant cystatin from Eudiplozoon nipponicum (Monogenea) modulates the cytokine production by macrophages in vitro Jana Ilgová1*, Lenka Kavanová2, Jiří Salát2, Milan Gelnar1, Martin Kašný1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2Department of Immunology, Veterinary Research Institute, Brno, Czech Republic *[email protected]

Parasites use wide repertoire of compounds contained in their excretory-secretory (ES) products to manipulate the immune system of their host. The inhibition of inflammation, altering the movement of the host’s white blood cells, changing of cytokine expression and interfering with antigen processing and/or presentation are just some of the mechanisms employed by the parasites to ensure increased chances for survival and higher fitness of the parasite. Among molecules which are involved in such modulation of the immune system belong inhibitors of cysteine peptidases (also called cystatins). In our previous work we characterized type I cystatin of monogenean hematophagous ectoparasite Eudiplozoon nipponicum (EnStef) detectable within its ES products1. Unlike other type I cystatin it possess unique domain which allows inhibition of legumain, in addition to blocking of papain-like peptidases typical for this type of inhibitors. The absence of legumain in the ES products and crude extract of the worm2 led us to assume that EnStef may be able to inhibit the legumain of the host origin. Since legumain as well as cysteine peptidases take part in many immune defense mechanisms of vertebrates, we were motivated to uncover the effect of recombinant EnStef (rEnStef) on the activity of fish leucocytes in vitro. Prior to in vitro tests we designed primers and optimized qPCR conditions for the amplification of carp’s reference genes (actin-beta, 40S ribosomal protein) and cytokine genes of interest (TNF-alpha, IL1-beta, IL-6, IL-8, IL- 10). We isolated and purified leucocytes from spleen and pronephros of common carp on Percoll gradient and established primary in vitro cell cultures. During these experiments we analyzed the expression levels of pro- and anti-inflammatory cytokines of the host immune cells in the presence of recombinant type I cystatin using qPCR. Nevertheless, our attempts to increase the expression of cytokines of interest by LPS was unsuccessful in fish cell cultures and will be subjected to studies in the future. Therefore, we tested the effect of rEnStef on optimized cell cultures of porcine alveolar macrophages. rEnStef caused significant downregulation of the expression of TNF- alpha and IL-10 in LPS stimulated porcine macrophages after 6 hours in incubation. We did not observe any effect on the production of cytokines on the protein level measured by ELISA, probably due to short time of incubation of cells with the recombinant inhibitor. Our finding indicates possible role of this inhibitor in immunomodulation of the host.

Acknowledgement This study was supported by Czech Science Foundation projects (P505/12/G112 and GBP505/12/G112).

References 1. Ilgová J., Jedličková L., Dvořáková H., Benovics M., Mikeš L., Janda L., Vorel J., Roudnický P., Potěšil D., Zdráhal Z., Gelnar M., Kašný M. (2017). A novel type I cystatin of parasite origin with atypical legumain- binding domain. Scientific Reports 7: 17526. 2. Jedličková L., Dvořáková H., Kašný M., Ilgová J., Potěšil D., Zdráhal Z., Mikeš L. (2016) Major acid endopeptidases of the blood-feeding monogenean Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae). Parasitology 143(4): 494–506.

Round goby impact on native fish assemblages: a meta-analysis approach Michal Janáč1* 1Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic *[email protected]

One of the major problems in invasion biology is the lack of reliable data on the impact of non-native species on recipient systems. Ideally, reliable evidence of impact requires long-term monitoring data from multiple sites with different invasion histories; otherwise the impacts observed could be confounded with other biotic or abiotic effects occurring at the same time or merely represent short-term fluctuations. Indeed, a large proportion of present data suggesting invasive species impact may be biased in either of these ways. However, as impact studies are rarely conducted on the same invasion (i.e. they differ in time and place), they are rarely subjected to the same bias and, as such, general invasion patterns may be derived through an impact meta-analysis. The present study aimed to demonstrate a methodological concept to meta-analyse various impact data on an invasive species, using round goby (Neogobius melanostomus; Gobiidae, Pisces) impacts on fish assemblages as a model system. The literature search for studies containing quantifiable, in situ obtained information on trends in fish species abundance attributable to round goby presence resulted in a dataset of 154 cases (one species in one invasion study) from 27 studies. Each case was attributed with an effect score (-1 = negative impact, 0 = neutral, +1 = positive impact) and a reliability coefficient (1–10) determining how reliable the trend observed was according to subjective evaluation based on a set of objectively determined criteria. For each species i, an Impact Score (ranging from -1 to +1) was calculated 푛 ∑푗=1 퐸푓푓푒푐푡푖푗 ∗ 푅푒푙푖푎푏푖푙푖푡푦 퐶표푒푓푓푖푐푖푒푛푡푖푗 퐼푆푖 = 푛 ∑푗=1 푅푒푙푖푎푏푖푙푖푡푦 퐶표푒푓푓푖푐푖푒푛푡푖푗 The dataset was used to test for (i) effect of continent, water body type and time since invasion on the effect observed (ordinal logistic regression with ‘species’ and ‘invasion study’ as random effects) and (ii) effect of target species size and ecotype on its impact score (linear regression). The study provides a clear evidence for case-specific impact of round goby: out of the ten target species with ≥4 records, none had all records pointing in the same direction (i.e. negative, neutral or positive impact). Generally, negative impacts prevailed in cottids, gobiids and darters. Smaller fish species tended to experience negative impact more commonly than larger fish, with no difference observed among the ecotype groups (demersal, benthopelagic and pelagic species). Round goby impact on North American fish assemblages was similar to that in Europe. Similarly, there was no difference between round goby impact in lentic and lotic waters.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

Structural and molecular diversity of dactylogyrids parasitizing African characiform fishes Maria L. Červenka Kičinja1*, Mária Seifertová1, Milan Gelnar1, Eva Řehulková1 Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

In 2018, my research activities continue to investigate the diversity of dactylogyrids parasitizing African tetras. Herein, the most important results are briefly summarized. Morphological and molecular characterizations of seven (three new) species of Characidotrema (C. auritum n. sp., C. brevipenis, C. nursei, C. pollex n. sp., C. spinivaginus, C. vespertilio n. sp., and C. zelotes) from Brycinus nurse and B. imberi have been provided [1]. Morphological analysis confirmed that the most apparent character distinguishing species in the genus is morphology of the male copulatory organ and vagina. Observations on the haptoral sclerotized structures of these parasites by employing phase contrast microscopy revealed the presence of an accessory structure relating to the ventral anchor, a feature that improves generic diagnosis of Characidotrema. Phylogenetic analyses based on 28S rDNA sequences supported the monophyly of Characidotrema spp., and indicated a closer relationship of this genus to monogeneans parasitizing African cyprinids (Dactylogyrus spp.) and cichlids (species of Cichlidogyrus, Scutogyrus, and Onchobdella) rather than to those parasitizing catfishes (species of Quadriacanthus, Schilbetrema, and Synodontella). The overall agreement between the morphological diversification of the MCOs and molecular tree observed in our study indicates that significant phylogenetic signals for clarifying relationships among species of Characidotrema are present in characteristics of the MCO. Morphological and molecular evaluation of 16 species of Annulotrema collected from nine species of African tetras (Alestes baremoze, A. dentex, Brycinus imberi, B. nurse, Hydrocynus forskahlii, H. brevis, H. vittatus, Micralestes acutidens, and M. elongatus) revealed the presence of nine undescribed species. The obtained molecular data will be used for phylogenetic reconstruction of monogeneans parasitizing African tetras. Finally, preliminary phylogenetic analyses of monogeneans parasitizing alestids showed that Afrocleidodiscus hydrocynuous from Hydrocynus forskahlii and representatives of Annulotrema clustered together, while A. paracleidodiscus from Distichodus rostratus (Characiformes: Distichodontidae) forms separate cluster close to the Characidotrema species. Division of the both species of Afrocleidodiscus highlights the necessity of a revision of the genus [2].

Acknowledgement This research was supported by the Czech Science Foundation No. P505/12/G112 (ECIP). A special thank is due to Iva Přikrylová, Maarten Vanhove, Eva Decru, Kateřina Francová, Michal Benovics, Matej Polačik and Radim Blažek for collecting monogeneans and/or their hosts.

References 1. Kičinjaová M.L., Seifertová M., Gelnar, M., Řehulková, E. Characidotrema species (Monogenea: Dactylogyridae) from the gills of Brycinus imberi and Brycinus nurse (Characiformes: Alestidae): new records, new species and first insights into the molecular phylogeny of the genus. [Submitted, Parasites and Vectors]. 2. Červenka Kičinja M.L., Seifertová M., Gelnar M., Řehulková E. Phylogenetic relationships among dactylogyrid species from African tetras. XIIIth Slovak and Czech parasitological days, 21.–25. May 2018, Košice, Slovak Republic. [Oral presentation].

Monogenean parasites of pelagic fish species in Lake Tanganyika: potential tags for host history and population structure Nikol Kmentová1*, Maarten Van Steenberge2,3, Stephan Koblmüller4, Tom Artois7, Fidel Muterezi Bukinga5, Theophile Mulimbwa N'sibula5, Pascal Masilya Mulungula5, Milan Gelnar1, Maarten P.M. Vanhove1,2,3,6,7 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2Biology Department, Royal Museum for Central Africa, Tervuren, Belgium; 3Operational Directorate Taxonomy and Phylogeny, Royal Belgian Institute of Natural Sciences, Brussels, Belgium; 4Institute of Zoology, University of Graz, Austria; 5Centre de Recherche en Hydrobiologie, Département de Biologie, Uvira, Democratic Republic of Congo; 6 Laboratory of Biodiversity and Evolutionary Genomics, Department of Biology, University of Leuven, Belgium; 7Centre for Environmental Sciences, Research Group Zoology: Biodiversity and Toxicology, Hasselt University, Diepenbeek, Belgium *[email protected]

Lake Tanganyika is the deepest of the African Great Lakes and harbours one of the most diverse fish assemblages. While dozens of studies focus on this lake’s cichlids as model organisms, our knowledge about the economically important fish species is still poor. The fishing effort is concentrated mainly in the lake’s pelagic zone with two clupeid (Limnothrissa miodon, Stolothrissa tanganicae) and four latid species (Lates angustifrons, L. mariae, L. microlepis, L. stappersii) as dominant targets. Additionally, cichlid species including members of the tribe Bathybatini can be found as valuable catch on fish markets. As abovementioned host taxa are known to be infected by monogenean parasites from three different genera [1,2], we examined additional fish specimens to explore 1) the parasites’ potential as tags to reveal their hosts’ population structure or history and 2) the origin of these freshwater parasites on these clupeid and latid hosts of marine origin. Samples of 14 host species (representatives of Cichlidae, Clupeidae, Latidae) originated from localities throughout Lake Tanganyika including all three subbasins of the lake. Parasite species identification was based on the sclerotised haptoral and genital structures. Intraspecific differences were analysed using morphometrics and geomorphometrics. Molecular characterisation was conducted using a range of nuclear and mitochondrial markers with different rates of molecular evolution. Based on phylogenetic reconstruction, the origin of diplectanids in African freshwaters is probably connected with latids diverging into African and Asian lineages. Parasite population structure inferred from part of the COI gene shows no north-south gradient. Correspondingly, no unique morphotype related to subbasin origin was reported neither for Kapentagyrus spp., nor Diplectanum lacustre. However, shape variation related to geographic origin inferred from geomorphometric analyses of Cichlidogyrus casuarinus, a monogenean infecting bathybatine cichlids, could be a sign of limited host migration. Therefore, the existence of parasite morphotypes related to the geographic origin of the hosts supports the possibility of using monogeneans as tags for population structure. Interestingly, morphological analyses of K. tanganicanus indicated that phenotypic variation also depended on host species. Similarly, our morphometric and geomorphometric analyses on C. casuarinus show some differentiation influenced by host preference. This pattern observed in different monogenean genera is probably caused by phenotypic changes during ontogenetic development because of its independence of genetic population structure. Recent demographic expansion in species infecting clupeid and cichlid hosts was detected and can be linked with paleogeographic events and climate change, respectively.

Future plans Geographic structure of Kapentagyrus spp and Cichlidogyrus casuarinus in Lake Tanganyika is planned to be analysed further by population genomics techniques.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); project No. P505/12/G112. The authors are grateful to Tine Huyse, Jos Snoeks, Miguël Parrent, Filip Volckaert and people in the parasitological group, Masaryk University, Brno and Research Centre of Hydrobiology, Uvira for their hospitality.

References 1. Kmentová N., Gelnar M., Mendlová M., Van Steenberge M., Koblmüller S., Vanhove M.P.M. (2016) Reduced host-specificity in a parasite infecting non-littoral Lake Tanganyika cichlids evidenced by intraspecific morphological and genetic diversity. Scientific Reports 6: 39605. 2. Kmentová N., Van Steenberge M., Raeymaekers J., et al. (2018) Monogenean parasites of sardines in Lake Tanganyika: diversity, origin and intra-specific variability. Contribution to Zoology 87: 105–132.

Fight with a jellyfish: host-parasite interaction of common parp and the myxozoan Sphaerospora molnari Tomáš Korytář1,2*, Sneha Patra1, Hana Pecková1, Astrid S. Holzer1 1Institute of Parasitology, Biology Center of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Faculty of Fisheries and Protection of Waters, University of South Bohemia, České Budějovice, Czech Republic *[email protected]

Sphaerospora molnari is myxozoan infecting common carp in central Europe, causing severe brachial lesions and necrosis. Prior to spore formation in the mucosal surfaces of the gill and skin, multicellular proliferative stages of S. molnari circulate for several weeks in the vascular system of its host [1]. Using our unique laboratory model of fish-to-fish injection of proliferative stages of S. molnari, parasite proliferation in different organs was analyzed over 63 days and correlated with changes in the blood composition and expression of selected cytokines. The obtained results identified two peaks of acute parasitemia on day 28 and 42 respectively. Unexpectedly, the highest parasite load was detected in the liver, a previously unknown localization of S. molnari. On a molecular level, the infection induced dynamic changes in the expression of pro- and anti- inflammatory cytokines, with a predominant role of IL-10. The hematological analysis observed a steady increase in the number of lymphocytes from day 28 onwards, correlating with the growing number of parasites, and only marginal changes in other populations. The analysis of IgM expression suggested an increase in the number of B cells throughout the course of infection, confirmed by flow cytometry. The increase of membrane-bound IgM was followed by a substantial increase in the expression of secretory IgM and increase of S. molnari-specific antibodies in the serum of infected fish. Overall, the obtained results provide the first insights into the kinetics of early infection host-parasite interaction and demonstrate the acquisition of S. molnari specific immunity [2].

Future plans Preparation and finalisation of related publication, identification of immune evasion strategies and molecules associated with this.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

References 1. Hartigan A., Estensoro I., Vancová M., Bílý T., Patra S., Eszterbauer E., Holzer A.S. (2016) New cell motility model observed in parasitic cnidarian Sphaerospora molnari (Myxozoa: Myxosporea) blood stages in fish. Scientific Reports 6: 39039. 2. Korytar T., Wiegertjes G., Zusková E., Tomanová A., Lisnerová A., Patra S., Sieranski V., Šíma R., Born-Torrijos A., Wentzel A., Yanes-Roca C., Astrid S. Holzer. The kinetics of cellular and humoral immune response mechanisms of common carp to presporogonic development of the myxozoan Sphaerospora molnari. [Submitted]

Cytoskeleton and motility in the archigregarine Selenidium pygospionis (Apicomplexa) Magdaléna Kováčiková1*, Andrei Diakin1, Andrea Valigurová1 1 Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

Based on their pleomorphic features, archigregarines are considered to represent the most ancestral gregarines or even apicomplexans as a whole [1]. Trophozoites of Selenidium pygospionis from marine polychaete Pygospio elegans Claparède, 1863 are covered by a typical apicomplexan three-layered pellicle (composed of the plasma membrane and inner membrane complex) that is folded into 28 broad and low folds separated by grooves [2]. The pellicle is underlain by longitudinally oriented subpellicular microtubules, arranged in a single layer, the continuity of which is interrupted only under the grooves where micropores are situated [2]. The role of subpellicular microtubules on bending motility performed by trophozoites of S. pygospionis in cooperation with actomyosin motor (Fig 1) heave been studied by different approaches (experimental motility assay on archigregarines isolated from host intestine, combination of light, electron and confocal microscopic analyses). In experimentally affected archigregarines treated with actin-modifying (jasplakinolide, cytochalasin D) or microtubule-destroying (oryzalin and colchicine) drugs, suppression or complete cessation of motility was documented. Electron microscopic analyses did not show any significant ultrastructural changes after jasplakinolide or cytochalasin D treatment. On the contrary, an obvious disappearance of normally continuous outermost microtubule set was observed in archigregarines incubated with drugs causing tubulin degradation (depolymerisation). This observation is in accordance with modifications of motility pattern monitored by the light microscopy and is additionally supported by results from confocal laser scanning microscopy, where immunolabelling revealed the presence of tubulin clusters instead of compact microtubules.

Future plans Submission of the manuscript.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112. Authors acknowledge Gita G. Paskerova (Saint-Petersburg State University), Timur G. Simdyanov (Moscow State University) and Naděžda Vaškovicová (Czech Academy of Sciences) for their help in field sampling and material analysis.

References 1. Leander B.S. (2007) Marine Gregarines: Evolutionary Prelude to the Apicomplexan Radiation? Trends in Parasitology 24(2): 60–67. 2. Paskerova, G.G., Miroliubova, T.S., Diakin, A., Kováčiková, M., Valigurová, A., Gillou, L., Aleoshin, V.V., Simdyanov, T.G. (2018) Fine structure and molecular phylogeny of two marine gregarines, Selenidium pygospionis n. sp. and S. pherusae n. sp., with notes on the phylogeny of Archigregarinida (Apicomplexa). Protist (in press).

Fig 1. Immunofluorescent labelling of actin (green) and myosin (red) in S. pygospionis (Scale: 20 μm)

The consequences of hybridization on metazoan parasite infection level in cyprinids Vadym Krasnovyd1*, Lukáš Vetešník2, Andrea Šimková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic 2Institute of Vertebrate, Biology of the Czech Academy of Sciences, Brno, Czech Republic *[email protected]

The hybrids are considered as a bridge-gap between parental species allowing parasite transfer from one host species to another. It was hypothesized that interruption of species-specific host-parasite co-adaptation may result in differences in the parasite load between parental species and their hybrids. In addition, it was also proposed that the maternal ancestry of the hybrids may contribute to the differences in parasite load between different hybrid lines. The main aim of this study was to investigate differences in parasite infection, and immune and physiological parameters between parental species and F1 hybrids using the selected cyprinid systems exhibiting hybridization. Additionally, we analyzed the effect of maternal origin of hybrids on parasite infection. Parasite infection level was examined in two cyprinid systems exhibiting hybridization. The first system represented the natural hybridization between two phylogenetically distant species, roach (Rutilus rutilus) and common bream (Abramis brama). All specimens were identified using morphological characteristics and genetic markers (partial mitochondrial cyt b gene and 12 microsatellite loci). The second system was represented by an artificially prepared lines of pure species and F1 hybrids using two phylogenetically closely related species, silver bream (Blicca bjoerkna) and common bream (A. brama). Monogeneans were dominant in the parasite communities of both pure species and their hybrids. In both systems, F1 hybrids displayed higher parasite species richness than their respective parental species, with both specialist and generalist parasites of the pure species present in the parasite communities of hybrids. However, some species-specific parasite species were not present in hybrids. On the other hand, parasite abundance was higher in parental species compared to the hybrids. We observed asymmetry in the distribution of specific parasites in hybrids in favour of the specific parasites of one parental species in both systems studied. In the system of phylogenetically distant cyprinid species, we detected a significant effect of maternal origin on the digenean and crustacean infection of hybrids. The significant effects of season, sampling site and year of collection on the composition of the metazoan parasite communities were found. In the system of phylogenetically distant cyprinid species, similarities in spleen- somatic index between hybrids and roach and in hepato-somatic index between hybrids and common bream were detected, nevertheless, gonado-somatic index in hybrids was intermediate between parental species. The non- specific immunity was affected only by season. The effects of the fish group (considering roach, common bream and hybrids) and sex on the fish condition were found. Our findings concerning the distribution of parental species-specific parasites in hybrids suggest that different host specific parasites display the different degrees of host-parasite co-adaptation. Our study showed that the hybrid’s protective immunological mechanism is more closely resemble that of one of the parental species.

Acknowledgement This study was funded by Czech Science Foundation, project no. P505/12/0375. From 2017, Vadym Krasnovyd and Andrea Šimková were partially funded through ECIP - Centre of Excellence, project no. P505/12/G112.

Asian fish tapeworm: the most successful invasive parasite in freshwaters Roman Kuchta1, Anindo Choudhury2, Tomáš Scholz1* 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Division of Natural Sciences, St. Norbert College, DePere, Wisconsin, USA *[email protected]

The Asian fish tapeworm, Schyzocotyle acheilognathi (Cestoda: Bothriocephalidea), is a notorious and highly successful invasive parasite reported in a wide spectrum of freshwater fishes, and new reports of its spread continue to emerge, especially in North America. To date, no thorough review of its world-wide distribution and host associations is available. In the present work, we use information from 651 articles up until 2017, and provide a summary. We increase the number of reported hosts to 312 fish species. One third of all records are from common and grass carp. In addition, 11 non-fish species may also harbour this parasite, which is quite unusual among fish helminths. The Asian fish tapeworm has spread to all but one (Antarctica) continents. The highest number of the records are from North America, followed by Asia and Europe. A key feature of its invasive success is its broad environmental tolerance (Kuchta et al., 2018).

Acknowledgement This review was financially supported by the Czech Science Foundation (project No. 15-14198S) and the Institute of Parasitology, BC CAS (project No. 60077344).

Reference 1. Kuchta R., Choudhury A., Scholz T. (2018) Asian fish tapeworm: the most successful invasive parasite in freshwaters. Trends in Parasitology 34: 511–523.

Not as reduced as we thought: identification of nematocyst protein NOWA in polar capsules of Myxozoa Jiří Kyslík1,2*, Anush Kosakyan1, Ivan Fiala1,2 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Department of Parasitology, University of South Bohemia, Faculty of Science, České Budějovice, Czech Republic *[email protected].

Cnidarians possess unique extrusive complex organelles, nematocysts, involved in the defense and capture of prey. Myxozoans as parasitic cnidarians evolved simplified homologues (!!!) of these cells called polar capsules. Myxozoans use these subcellular organelles in their spore stages for the attachment to the host. Recently conducted genomic and functional analyses identified several nematocyst-specific genes which play a critical role in morphogenesis and evolution of cnidarian nematocysts [1,2]. Family of unusually short collagens, termed minicollagens, are the major constituent of the nematocyst capsule wall. Proteome and genome of cnidarian model Hydra showed a high diversity of minicollagens (17 types) [3]. Generally, minicollagens comprise a central collagen triple helix with 12–16 Gly-X-Y repeats flanked by polyproline stretches and terminal cysteine-rich domains (CRDs) with a conserved pattern of six closely set cysteines. Homologous cysteine-rich sequence composition with C-terminal octarepeat domain of CRDs was observed in glycoprotein with globular structure in outer surface of nematocysts (NOWA; nematocyst outer wall antigen) [4]. NOWA serves as a positional organizer of minicollagen assembly in highly compacted suprastructure using disulfide-dependent heteroassembly of CRDs with minicollagens. Hence, NOWA mediates cross-linking of minicollagens to networks in the inner wall of the capsule. Apart from the eightfold repeated cysteine-rich domain, NOWA protein possesses N-terminal sperm coating glycoprotein domain (SCP) and a central C-type lectin-like domain (CTLD). Genome and transcriptome studies on specific polar capsule genes surprisingly did not reveal any homolog of NOWA [5]. Clarifying the molecular composition of myxozoan polar capsules represents a key for understanding the evolutionary origins of polar capsules of these reduced jellyfish. Here we present an identification of NOWA homologs in Myxozoa using bioinformatic analysis of next-gen data (large-scale genomic comparison, reciprocal BLAST etc.). Putative NOWA transcripts show domain reduction and variable amino acid composition, which may be associated with fast evolution of Myxozoa and their morphological reduction. We successfully identified potential NOWA homologs in genomic and transcriptomic data of 7 myxozoan species including our obtained transcriptomic data of Myxidium lieberkuehni. Domain architecture of myxozoan NOWA transcripts shows significant reduction from octarepeat cysteine-rich domain to 4-5 repeats with conserved C-terminal part. Nevertheless, amino acid composition within the C-terminal part possesses variable motif between freshwater and marine species suggesting conformational changes. However several domain loses were observed. Myxozoa lack two NOWA domains (SCP, CTLD) and thus their sequence length is truncated compared to cnidarian homologs. Differential expression analysis of the transcriptome of Sphaerospora molnari using DESeq [6] revealed the high expression value of NOWA transcript in spore-forming stages compared to the blood stages, which supports our suggestion of correct identification of myxozoan NOWA proteins. Identification of polar capsule proteins and their role in the development of polar capsules can bring a significant step in the knowledge of the evolution of these organelles and in the future development of potential drug targets.

Future plans Quantitative real-time expression analysis of identified NOWA transcript in M. lieberkuehni. Preparation of manuscript for future publication concerning obtained data.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

References 1. Milde S., Hemmrich G., Anton-Erxleben F., Khalturin K., Wittlieb J., Bosch T.G.G. (2009) Characterization of taxonomically restricted genes in a phylum-restricted cell type. Genome Biology 10: R8. 2. Hwang J.S., Takaku Y., Momose T., Adamczyk P., Özbek S., Ikeo K., Khalturin K., Hemmrich G., Bosch T.C., Holstein T.W., David C.N., Gojobori T. (2010) Nematogalectin, a nematocyst protein with GlyXY and galectin domains, demonstrates nematocyte-specific alternative splicing in Hydra. Proceedings of the National Academy of Sciences of the United States of America 107: 18539–18544.

3. David C.N., Özbek S., Adamczyk P., Meier S., Pauly B., Chapman J., Hwang J.S., Gojobori T., Holstein T.W. (2008) Evolution of complex structures: minicollagens shape the cnidarian nematocyst. Trends in Genetics 28: 431–438 4. Özbek S., Pokidysheva E., Schwager M., Schulthess T., Tariq N., Barth D., Milbradt A.G., Moroder L., Engel J., Holstein T.W. (2004) The Glycoprotein NOWA and Minicollagens Are Part of a Disulfidelinked Polymer That Forms the Cnidarian Nematocyst Wall. The Journal of Biological Chemistry 279: 52016–52023. 5. Shpirer E., Chang E.S., Diamant A., Rubinstein N., Cartwright P., Huchon D. (2014) Diversity and evolution of myxozoan minicollagens and nematogalectins. BMC Evolutionary Biology 14. 6. Anders S, Huber W (2010) “Differential expression analysis for sequence count data.” Genome Biology 11: R106.

Myxozoa wherever you look: Uncovering myxozoan species diversity Martina Lisnerová1,2, Ivan Fiala1,2* 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice 2Faculty of Sciences, University of South Bohemia, České Budějovice *[email protected]

Myxozoans are microscopic metazoan parasites infecting typically fish as alternate hosts and annelids as definitive hosts. Some myxozoans are economically important parasites causing serious fish diseases. There are about 2300 described species of these morphologically extremely reduced cnidarian parasites classified in 67 genera. However, myxozoan species diversity still remains highly unrecognized. Some authors estimate that only in neotropical region there might be up to 16000 myxosporean species due to high diversity of freshwater fish in the Amazon river region and high host specificity of myxosporean species. We performed myxozoan screening in freshwater fish from selected ponds, rivers and dams in south and central Bohemia to assess a myxozoan biodiversity in the region that has a long term history of myxozoan research and most of the known myxozoans are already characterized by 18S rDNA sequences. We screened 30 fish species (285 fish individuals) from 18 different localities. We performed light microscopy and PCR screening of mostly gills, kidneys and gall bladders, typical sites of myxosporean infection. By SSU rDNA sequencing we revealed 53 different myxosporean species. Seventeen SSU rDNA sequences were identical with sequences available in GenBank and 36 sequences belong to newly identified myxozoan taxa. Interestingly, not only large dams as Římov and Švihov contain high number of myxozoans (e.g. 19 myxosporeans in 8 screened fish species in Švihov) but also a small brook, Hostačovka is home for 9 myxosporeans based only on three screened fish species, or six fish species from a very small pond are host for 16 myxosporean species. Rutilus rutilus was the most infected fish species with 8 recognised myxosporeans.

Future plans Diversity of the Myxozoa is highly underestimated even in the areas of long tradition of myxozoan research. We are currently working on environmental DNA sequencing trying to detect DNA of myxozoan spores in the water and water sediments that would enable us more efficiently assess the true myxozoan diversity.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

Rasheedia n. nom. (Nematoda: Physalopteridae) for Bulbocephalus Rasheed, 1966 (a homonym of Bulbocephalus Watson, 1916), with description of Rasheedia heptacanthi n. sp. and R. novaecaledoniensis n. sp. from perciform fishes off New Caledonia František Moravec1*, Jean-Lou Justine2 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Institut Systématique Évolution Biodiversité, Muséum National d’Histoire Naturelle, CNRS, Sorbonne Université, Paris, France *[email protected]

The nematode genus Bulbocephalus Rasheed, 1966 (Nematoda: Physalopteridae) was found to be a homonym of Bulbocephalus Watson, 1916 (Apicomplexa) and, therefore, a new name, Rasheedia n. nom., is proposed to substitute it. Based on light and scanning electron microscopical studies of nematode specimens collected from the digestive tract of perciform fishes off New Caledonia, two new species of Rasheedia are reported: R. heptacanthi n. sp. from the Cinnabar goatfish Parupeneus heptacanthus (Lacépède) (Mullidae) (type host) and Dentex fourmanoiri Akazaki and Séret (Sparidae), and R. novaecaledoniensis n. sp. from the Indian goatfish Parupeneus indicus (Shaw) (Mullidae). These new species are mainly characterized by the number of anterior protrusible oesophageal lobes (two in R. heptacanthi and four in R. novaecaledoniensis), structure of the oesophagus and the lengths of spicules. An amended diagnosis of Rasheedia and a key to species of this genus are provided. Three previously described congeneric species are transferred to Rasheedia as R. deblocki (Le-Van-Hoa, Pham-Ngog-Khue and Nguyen-Thi-Lien, 1972) n. comb., R. inglisi (Rasheed, 1966) n. comb. and R. pseudupenei (Vassiliadès and Diaw, 1978) n. comb. Cestocephalus Rasheed, 1966 [genus inquirendum], including C. serratus Rasheed, 1966 and C. petterae (Le-Van-Hoa, Pham-Ngog-Khue and Nguyen-Thi-Lien, 1972) n. comb., should be considered to be separate from Rasheedia. The names Pseudomazzia Bilqees, Ghazi and Haseeb, 2005 and P. macrolabiata Bilqees, Ghazi and Haseeb, 2005, established for a nematode somewhat resembling Rasheedia spp., should be considered nomina dubia. Rasheedia heptacanthi n. sp. and R. novaecaledoniensis n. sp. are the first representatives of the family Physalopteridae recorded from fishes in New Caledonian waters.

Future plans Further studies of the morphology and taxonomy of nematodes parasitizing freshwater and marine fishes in the conditions of different geographical regions.

Acknowledgement This study was partly supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

References 1. Rasheed S. (1966) On some interesting nematode parasites of fish from Pakistan. Parasitology 56(1): 151– 160. 2. Watson M.E. (1916) Observations on polycystid gregarines from Arthropoda. Journal of Parasitology 3(2): 65–75.

Effects of the infection of eye fluke Diplostomum pseudospathaceum on reproductive traits and metabolism of European bitterling Rhodeus amarus Veronika Nezhybová1,2*, Martin Reichard1, Caroline Methling1, Andrea Slaninová1, Markéta Ondračková1 1Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic 2Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

In semi-experimental conditions an extensive study was conducted to determine the effects of the infection with the metacercariae of Diplostomum pseudospathaceum on several aspects of reproduction of its second host, the European bitterling, Rhodeus amarus. The size and colour intensity of the red spot in male´s iris, an important sexual trait, differed between infected and control males with control males having generally smaller but more pronounced spot than Diplostomum-infected males. As the intensity of red colouration in males is considered as an important trait for reproductive success, these results suggest an advantage of control males in gaining the favour of females over infected males. The number of females with developed ovipositor and the number of produced offspring were comparable between infected and control group but there was one to two-week delay in infected female reproduction peak and offspring release. Siring the offspring later in the season could be disadvantageous for infected females because of possible higher mortality of embryos in mussels and shorter time for gaining the energetic supplies before the overwintering. However, both standard and maximum metabolic rate measured as oxygen consumption did not differ between infected and control groups suggesting minimal or no costs of the chronic (several months) infection on bitterling metabolism. During behavioural observation of intersexual selection, infected and control males and females did not significantly differ in their reproductive activity which corresponds with the results of metabolic rate. When determining the female preference for infected or control males, there was no significant difference when both visual and olfactory contact between females and males was allowed. However, fish tended to respond more intensively to partners of the same health status when olfactory contact was prevented. In all trials, male activity significantly affected the behaviour of responding female irrespectively of the health status, and oppositely. To summarize, the impact of the infection by D. pseudospathaceum on the reproduction of European bitterling was weaker than expected. However, experimental fish were held under favourable conditions with sufficient food intake and breeding substrate (mussels) and without predators and other infections and, therefore, the effects of this parasite is suggested to be more pronounced in natural conditions.

Future plans Finalisation of publication concerning the data in presented abstract and manuscripts “Morphological and molecular study on nematode larvae of killifish (Nothobranchiidae) from Mozambique, Africa” (autors: V. Nezhybová, M. Seifertová, M. Ondračková, Š. Mašová) and “Parasite-induced alteration of the behaviour of fish host Nothobranchius furzeri from Mozambique: the effect of natural Apatemon sp. infection” (autors: V. Nezhybová, M. Janáč, M. Reichard, M. Ondračková).

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

Parasite acquisition by non-native centrarchid fish Lepomis gibbosus in Europe Markéta Ondračková1*, Yuriy Kvach1,2, Mária Seifertová3, Jitka Fojtů1, Markéta Pravdová1,2, Pavel Jurajda1 1Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic; 2 Institute of Marine Biology of NAS of Ukraine, Odessa, Ukraine; 3Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

Metazoan parasite communities of pumpkinseed sunfish Lepomis gibbosus (Centrarchidae), one of the most successfully introduced fish species in Europe, were studied at two isolated ponds (Knielingen, Tropfen) along the Upper Rhine in Germany. Nine parasite taxa were observed, including North-American species co-introduced to Europe (ancyrocephalid monogeneans, diplostomid trematodes), circumpolar species infecting L. gibbosus in both their native and non-native ranges (bothriocephalid cestodes) and locally acquired parasitic nematodes Camallanus lacustris and Contracaecum ovale. Acquisition of local parasites was not observed at Tropfen, where the fish community comprised just two species, with L. gibbosus dominant. Low prevalence and abundance of acquired parasites was found at Knielingen, which supported a diverse fish community. Due to low local parasite competency, L. gibbosus appears to have no significant impact on parasite dynamics in sites investigated in this study. Susceptibility to local, but also non-native parasite species was investigated in the floodplain of the Dyje River, Danube basin, Czech Republic. The parasitic copepod Neoergasilus japonicus (Harada, 1930) (Ergasilidae), native to East Asia, is widely distributed in Asia, Europe, and North and Central America. Recently this species appeared in lentic water bodies of the River Dyje floodplain. It was first recorded in 2015 and in two years it reached 100% prevalence in recently expanding non-native fish host, Lepomis gibbosus, native to North-America, at two borrow pits. Abundance of N. japonicus increased with fish length, with maximum intensity of infection reaching 99 parasites per fish. The parasite was most frequently found attached to the dorsal and anal fins of fish, while preference for the dorsal fin was more evident with lower infection intensities. Utilization of expanding fish hosts in water bodies that are regularly interconnected via natural or managed flooding may support the rapid dispersal of this non-native parasite.

Future plans Identification of parasites collected in France, Austria and the Czech Republic, data analysis and preparation of manuscripts regarding parasite communities of Lepomis gibbosus in Europe.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

A new classification of Glaridacris Cooper, 1920 (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in North America, including erection of Pseudoglaridacris n. gen. Mikuláš Oros1, Dalibor Uhrovič1, Tomáš Scholz2* 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Institute of Parasitology, Slovak Academy of Sciences, Košice, Slovakia *[email protected]

A taxonomic study of monozoic cestodes of the genus Glaridacris Cooper, 1920 (Cestoda: Caryophyllidea), parasites of catostomid fishes in North America, confirmed artificial character of the genus which is split to two different, morphologically distinct and not closely related genera. Glaridacris is newly circumscribed to include only three species, Glaridacris catostomi Cooper, 1920 (type species), Glaridacris terebrans (Linton, 1893) and Glaridacris vogei Mackiewicz, 1976, which are characterised by an elongate body, a cuneiloculate or wedge-shaped scolex with six shallow loculi, male and female gonopores at a distance from each other, follicular ovary and circum-medullary vitelline follicles (lateral and median). A new genus Psedoglaridacris n. gen. is proposed to accommodate three species characterised by a shorter body, a bothrioloculodiscate scolex with a pair of deeper median bothria and two shallower loculi, male and female gonopores close together, non-follicular ovary and with only lateral vitelline follicles. The species are: Pseudoglaridacris laruei (Lamont, 1921) n. comb. (type species), Pseudoglaridacris confusa (Hunter, 1929) n. comb., and Pseudoglaridacris oligorchis (Haderlie, 1953) n. comb. An annotated list of all species of both genera with data on their hosts and distribution, and keys to their identification are provided (Oros et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Oros M., Uhrovič, Scholz T. (2018) A new classification of Glaridacris Cooper, 1920 (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in North America, including erection of Pseudoglaridacris n. gen. Journal of Parasitology 104: 60–69.

Factors contributing to the coexistence of two reproductive forms of Carassius gibelio in the Czech Republic. Pakosta Tomáš1*, Lukáš Vetešník2, Pavel Hyršl3, Andrea Šimková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic; 3Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

The first records of Carassius auratus gibelio species arose in 1976. Since 1990 the transformation from originally triploid (likely gynogenetic) towards the mixed diploid-polyploid population (with coexisting gynogenetic and sexual forms) was reported. This unique fish species, which originated from Asia, is characterized by high environmental tolerance and adaptability. Therefore, it is a perfect model of a successful, invasive species. However, until now, the potential differences in physiology, immunity and/or parasitic between two coexisting reproductive forms were not fully investigated. The aim of this study was to investigate the parasite load and the selected physiological and immune parameters in two reproductive forms of C. auratus gibelio. According to mtDNA analysis, C. gibelio, which is a target of our study, is the most common form of Carassius auratus complex in the Czech Republic. Apart from C. auratus gibelio there are also other three forms reported in Czech Republic in low frequency i.e. C. auratus auratus, C. auratus langsdorfii and C. auratus M-line. In the frame of our four-year study, 22 different species of metazoan parasites were found on 195 fish individuals investigated. Monogenea represented the most frequent group of parasites in both diploid likely sexual females and males, and triploid likely gynogenetic females. From all parasite species, 3 species of Monogenea, namely in Dactylogyrus dulkeiti, Dactylogyrus anchoratus and Gyrodactylus sprostonae were found in the high prevalence and the highest intensity of infection. This pattern was recognized in both reproduction forms. We also found that both sexual individuals and gynogenetic females were parasitized more in the last year in comparison with three previous years. We did not confirm the prediction of Red Queen hypothesis as we did not find significantly higher level of infection in gynogenetic females compared to sexual forms in any of four years studied. Concerning physiological parameters studied, we confirmed statistically higher erythrocyte count in diploid form when compared to triploid form. On the other hand, higher level of haematocrit was found in diploid males compared to both groups of females. From the immunity point of view, we did not observe any significant difference between reproductive forms of C. auratus gibelio. Nevertheless, difference in some physiological and immune parameters were found among the specimens collected in different years. Activity of oxidative burst reached significantly lower values in the last year of study for both forms. On the other hand, in the same year the activity of the complement was significantly higher when compared to other 3 years. Condition factor and gonado-somatic factor were lower in diploid males when compared to sexual and asexual females. We also reported higher value of spleen-somatic index (considered as an indicator of immunocompetence) of diploid males when compared to both females. Thus, it seems that the high investment in gonad development in both gynogenetic and sexual females is compensated by low investment in immunocompetence. This result may indicate that investment into traits related to the maintenance, reproduction and immunity are determined rather by sex than by different ploidy level likely related to different modes of reproduction (gynogenetic vs. sexual). This may contribute to the coexistence of two reproduction forms of C. gibelio living in the same environment.

Acknowledgement This study was from 2017 supported by ECIP (European Centre of IchthyoParasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

References 1. Papoušek I. (2008) Molecular-genetic analyses of Carassius species in central Europe [Ph.D. thesis]. Masaryk University, Faculty of Science, Brno, Czech Republic. 2. Šimková A., Hyršl P., Halačka K., Vetešník L. (2015) Physiological and condition-related traits in the gynogenetic-sexual Carassius auratus complex: different investments promoting the coexistence of two reproductive forms? BMC Evolutionary Biology 15: 154 3. Kubala L., Lojek A., Číž M., Vondráček J., Dušková M., Slavíková H. (1996) Determination of phagocyte activity in whole blood of carp (Cyprinus carpio) by luminol-enhanced chemiluminescence. Veterinary Medicine 41(10): 323–327 4. Lusková V., Lusk S., Halačka K., Vetešník L. (2010) Carassius auratus gibelio – The most successful invasive fish in waters of the Czech Republic. Rusian Journal of Biological Invasions 1(3): 176–180.

New genera and species of paramphistomes (Digenea, Paramphistomoidea, Cladorchiidae) parasitic in fishes from the Amazon basin in Peru Camila Pantoja1, Tomáš Scholz2*, José Luis Luque1, Arlene Jones3 1Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil; 2Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 3Creigiau Llwydion, Heol y Maes, Coelbren, Neath, United Kingdoms *[email protected]

Two new genera and three new species of paramphistomoid digeneans are described in the family Cladorchiidae from doradid, heptapterid, pimelodid and pseudopimelodid fishes in the Amazon River in Peru. Goeldamphistomum amazonum n. g., n. sp. (type-species) from Goeldiella eques (type-host) and Tenellus trimaculatus, and Goeldamphistomum peruanum n. sp. from Duopalatinus peruanus (type-host), Calophysus macropterus and Microglanis sp. are placed in the Dadayiinae. Both species have an accessory acetabular sucker, which distinguishes the genus from all taxa previously reported from South American freshwater fishes. They differ from each other primarily in that G. amazonum has a prebifurcal genital pore and oblique, separated testes, the levels of which rarely overlap longitudinally, whereas G. peruanum has a postbifurcal genital pore and testes directly to obliquely tandem. Iquitostrema papillatum n. g., n. sp. (Kalitrematinae) from the intestine of Hassar orestis differs from other members of the subfamily in the combination of a massive acetabulum with a papillate luminal surface and symmetrical testes which overlie the caeca close to the caecal arch. These are the first records of paramphistomes from the five host- species studied here (Pantoja et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Pantoja C., Scholz T., Luque J.L., Jones A. (2018) New genera and species of paramphistomes (Digenea: Paramphistomoidea: Cladorchiidae) parasitic in fishes from the Amazon basin in Peru. Systematic Parasitology 95: 611–624.

Is the presence of pharmaceuticals in fish tissues associated with the abundance of parasites? Markéta Pravdová1,2*, Markéta Ondračková1, Jitka Kolářová3, Kateřina Grabicová3, Libor Mikl1, Martin Bláha3, Tomáš Randák3, Yurii Kvach1,4, Pavel Jurajda1 1Institute of vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic; 2Faculty of Science, Masaryk University Brno, Brno, Czech Republic; 3Faculty of Fisheries and Protection of Waters, South Bohemian Research Centre of Aquaculture and Biodiversity of Hydrocenoses, University of South Bohemia in České Budějovice, Czech Republic; 4Institute of Marine Biology of NAS of Ukraine, Odessa, Ukraine *[email protected]

Recently, there are many studies dealing with pollutants and their impact on the aquatic environment. One of the important stressors is represented by pharmaceuticals, which are naturally found near large cities, especially downstream sewage treatment plants. Due to the impossibility of separation of individual compounds, they form a complex mixture in nature resulting in several interactions. Pharmaceuticals usually affect immunity, physiology and behaviour of target organisms, as parasites do, and thus these stressors may interact with each other [1]. The aim of our study was to evaluate possible consequences of the presence of pharmaceuticals in fish tissues and possible impacts on the abundance of parasites. We sampled juvenile brown trout (Salmo trutta m. fario) and macrozoobenthic organisms in a small stream affected by effluent of municipal sewage treatment plant. Two sites, differing in concentrations of pharmaceuticals, were sampled; control, referred as unpolluted site in the sense of low pollution, and polluted site, located downstream effluent of sewage treatment plant with high concentrations of a number of pharmaceutical compounds. Results of the present study show ability of juvenile fish to accumulate high number of pharmaceutical compounds, despite their relatively low environmental concentrations. The highest concentrations were found for antidepressants and antibiotics of pharmaceuticals, being detected mainly in fish liver, followed by kidney and brain, where the same trend occurred at both sites. Fish condition was comparable between the sites, however, at polluted site, it tended to decrease with overall pharmaceutical load. Abundance of parasites differed between the sites, with higher abundance of ectoparasites, namely monogeneans, detected at polluted site, and higher abundance of endoparasitic species found at control site, irrespectively of the density of parasites´ intermediate hosts. Out of several medicinal classes detected in the fish tissues, psychopharmaceuticals seem to be important compounds potentially affecting the infection of the host by parasites, as concentration of antidepressants, anti-dementia drugs and antipsychotics significantly correlated with parasite load.

Future plans Complete the data analysis and preparation of manuscript.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No P505/12/G112.

References 1. Sures B., Nachev M., Selbach C., Marcogliese D.J. (2017) Parasite responses to pollution: what we know and where we go in “Environmental Parasitology“. Parasites & Vectors 10: 65.

Phenotypic plasticity in Cichlidogyrus spp. (Monogenea: Dactylogyridae) parasitizing Lake Tanganyika cichlid tribes using a geometric morphometric approach: the roles of host phylogeny and locality Chahrazed Rahmouni1*, Maarten Van Steenberge2,3,4, Maarten P.M. Vanhove1,4,5, Andrea Šimková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2Operational Direction Taxonomy and Phylogeny, Royal Belgian Institute for Natural Sciences, Brussels, Belgium; 3Section Vertebrates, Ichthyology, Royal Museum for Central Africa, Tervuren, Belgium; 4Laboratory of Biodiversity and Evolutionary Genomics, Department of Biology, University of Leuven, Leuven, Belgium; 5Centre for Environmental Sciences, Research Group Zoology: Biodiversity & Toxicology, Universiteit Hasselt, Diepenbeek, Belgium *[email protected]

Morphological plasticity is the ability of an organism to express different phenotypes depending on biotic and/or abiotic factors of the environment. A lot of consideration has been given to phenotypic plasticity in monogeneans. This group is, so far, the best model system for addressing fundamental ecological and evolutionary questions on parasites. The morphology of their haptor allows the attachment to the fish gills and is crucial for their survival. It also influences the parasite’s specialization and adaptation to the host species, and considerably contributes to host specificity. Variability of the sclerotized structures seems to be more pronounced in generalist species. African cichlids have been extensively studied for their monogenean parasite fauna. Host specificity in Cichlidogyrus Peperna, 1960, which represents the most diverse monogenean genus parasitizing African cichlids, ranges from strict specialists to generalists. Previous studies have led to some preliminary understanding of morphological evolution in Cichlidogyrus. To date, investigations about phenotypic plasticity in monogenean parasites involved only non- Tanganyikan cichlid hosts or deep-water species of the Tanganyika system. Our study aimed to determine whether or not conspecific species of Cichlidogyrus from different Lake Tanganyika cichlid tribes living in various localities exhibit differently shaped and sized dorsal and ventral anchors. A geomorphometric approache allows to investigate morphological plasticity patterns among populations of monogeneans in any ecological system and in a broad biogeographic and evolutionary context. We applied a landmark-based geomorphometric approach on four species of Cichlidogyrus sampled from various host species and from various localities along the Lake. The studied Cichlidogyrus species include i) a generalist species C. nshomboi isolated from representatives of two phylogenetically distinct cichlid tribes: Boulengerochromini represented by Boulengerochromis microlepis, and Perissodini by Perissodus microlepis and Plecodus straeleni; (ii) an intermediate specialist represented by C. gillardinae from the gills of congeneric hosts living in sympatry: Astatotilapia burtoni and A. stappersii (Haplochromini); (iii) a strict specialist represented by C. gistelincki from ’Ctenochromis‘ horei (Tropheini), a cichlid with good dispersal capacities of the shallow intermediate habitat; and finally (iv) C. milangelnari from Cyprichromis microlepidotus (Cyprichromini), a strict specialist of a poorly dispersing cichlid of the deep rocky waters. Geomorphometric analysis revealed significant differences in dorsal and ventral anchors between C. nshomboi individuals parasitizing the representatives of boulengerochromine and perissodine cichlids. The same result was observed between parasite individuals of perissodine hosts sampled from the Burundese and Congolese coastlines. However, no significant differences were found between individuals of C. nshomboi parasitizing P. microlepis and P. straeleni, both sampled from the Congolese shoreline. Significant differences in dorsal and ventral anchors were observed between individuals of C. milangelnari sampled from Burundese and Congolese parts of the lake, and C. gistelincki from Burundese localities. In C. gillardinae parasitizing two different congeneric hosts, significant differences were observed only in the dorsal anchors.

Future plans Preparation of two research papers focussed on geomorphometry of Cichlidogyrus parasites using the data set presented above, and one phylogenetic paper including molecular data presented in previous ECIP meetings. Finally, in cooperation with colleagues from the University of Basel, we will prepare a manuscript focussing on parasite species infecting haplochromine cichlids inhabiting Lake Tanganyika and some neighbouring freshwater systems.

Acknowledgement This study was supported by Czech Science Foundation, project No. P505/12/G112 (ECIP).

Concentrations of fourteen trace metals in scales of three nototheniid fishes from Antarctica (James Ross Island, Antarctic Peninsula) Kevin Roche1*, Jan Kuta2, Ivo Sedláček3, Rostislav Červenka2, Kateřina Tomanová3, Pavel Jurajda1 1Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic; 2Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic; 3Czech Collection of Microorganisms, Masaryk University, Faculty of Science, Brno, Czech Republic *[email protected]

Relatively few studies of trace metal contamination have been undertaken around Antarctica, with most taking place in areas of deep-water commercial fisheries or at sites affected by human activity. There is almost no information on trace metals in fishes from Antarctic coastal waters, despite their importance as feeding and breeding grounds. In this study, we assessed concentrations of 14 trace metals (Zn, Al, Mn, Fe, V, Cr, Co, Ni, Cu, As, Cd, Pb, Hg, U) in the scales of Notothenia coriiceps, Trematomus bernacchii and Gobionotothen gibberifrons caught off James Ross Island (Antarctic Peninsula). Overall, our results for scales broadly matched those of previous studies using different fish and different organs, with most metals found at trace levels (Hg always lowest) and Mn, Al, Fe and Zn occurring at high mean levels in all species (Zn always highest) [1,2]. High accumulation of Mn, Al, Fe and Zn is largely due to high levels in sediments associated with active volcanic sites. To the best of our knowledge, this study provides the first quantitative baseline data on U in coastal Antarctic fishes [1] and the first data on Al, Fe [2] and U [1] in nototheniid scales. Regressions of trace metal content against fish standard length indicated significant positive bioaccumulation of Mn, V, U and Al in T. bernacchii, along with non-significant positive accumulation of Fe, Zn, Co and Cr, most likely due to greater dietary specialisation sediment feeding benthic prey with calcareous shells [1,2]. Levels of significance were strongly affected by large-scale variation driven by differences in diet and/or seasonal changes in habitat use and sex differences associated with life stage and reproductive status. Levels of Mn, Al, Fe and Zn are being increased further by rising air borne deposition, also the primary source of V and U in this study. Increasing levels in ice core samples since the 1980s are attributed to increased mining and land use changes in South America and Australia. Further, increasing precipitation and temperatures associated with climate change have increased meltwater run off, thereby further increasing levels available to aquatic biota. Further long-term studies are encouraged to assess mechanisms of uptake (especially for Al) and possible intra- and interspecific impacts of climate change on the delicate Antarctic food web, recognising that the feeding ecology of individual species (and even individuals) can have a strong effect on overall trends.

Future plans This study has now ended and there are no further plans.

Acknowledgement This study was made possible thanks to financial support from the Grant Agency of the Czech Republic (ECIP Project No. P505/12/G112). We would like to express our thanks for permission to use the J.G. Mendel Czech Antarctic Station on James Ross Island in 2014 and all our expedition colleagues for their technical help while stationed there. Research activities at the J.G. Mendel Station are carried out as part of the RECETOX Research Infrastructure, financed through the Ministry of Education, Youth and Sports of the Czech Republic, project no. LM2015078, and European Structural and Investment Funds, Operational Programme for Research, Development and Education (CZ.02.1.01/0.0/0.0/16_013/0001761).

References 1. Roche K, Kuta J, Sedláček I, Červenka R, Tomanová K, Jurajda P (2018) First data on uranium uptake in three nototheniid fishes from Antarctica (James Ross Island). Chemosphere 211: 510–514. [Epub ahead of print] 2. Roche K, Kuta J, Sedláček I, Červenka R, Tomanová K, Jurajda P (2018) Concentrations of thirteen trace metals in scales of three nototheniid fishes from Antarctica (James Ross Island, Antarctic Peninsula). Biological Trace Element Research (in preparation)

Laser microdissection and mass spectrometry: proteomic tissue profiling of Eudiplozoon nipponicum Pavel Roudnický1*, Jiří Vorel1, Tomáš Loja2, David Potěšil3, Zbyněk Zdráhal3, Milan Gelnar1, Martin Kašný1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2CEITEC, Core Facility Genomics, Masaryk University, Brno, Czech Republic; 3CEITEC, Core Facility Proteomics, Masaryk University, Brno, Czech Republic *[email protected]

Laser microdissection is a combined microscopic-separation technique enabling to extract the samples of interest from histological slides - single cells, organs and tissues. The isolated samples can be further analysed and various parameters determined. The adoption of mass spectrometry (MS) analysis could give us the detail information about the sample composition and relative quantity of particular components (e.g. protein molecules). Moreover, this technology enables the fast determination of distribution of selected molecule within the studied biological object, such as e.g. Eudiplozoon nipponicum body. Thus, we can avoid the time-consuming preparation of specific antibodies in laboratory animals, which are widely used for protein tissue localization. During last year, we conducted experiments focused on generating the proteomic data of E. nipponicum specific tissues as a tool for localisation of proteins (e.g. serpin). We have identified 405 unique proteins in inner area of the body and 131 unique proteins in tegument. Among them, some already described functional protein molecules have been found (cathepsin L, B [1] and serpin).

Future plans Complete the collection of samples and run final MS analyses including biological and technical replicates. Evaluate obtained data and verify hypothesis, that serpin is present in the gut (regulation of digestion) of E. nipponicum and thus is released into the host where is potentially down regulating the inflammation. These results should be published in the second half of the year 2019.

Acknowledgement The research was financially supported by Czech Science Foundation (GBP505/12/G112) and Masaryk University (MUNI/A/0816/2017).

References 1. Jedličková L., Dvořáková H., Dvořák J., Kašný M., Ulrychová L., Vorel J., Žárský V., Mikeš L. (2018) Cysteine peptidases of Eudiplozoon nipponicum: a broad repertoire of structurally assorted cathepsins L in contrast to the scarcity of cathepsins B in an invasive species of haematophagous monogenean of common carp. Prasites & Vectors 11: 1–17.

Molecular characterization of gill monogeneans (Monogenea: Dactylogyridae) of African freshwater fishes Mária Seifertová1*, Kateřina Francová1, Maria L. Červenka Kičinja1, Eva Řehulková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

The DNA amplification through PCR and sequencing has been used for parasitic helminth diagnostics and phylogenetics for more than 20 y. So far most commonly used molecular markers in taxonomic classification of helminth parasites are different regions of nuclear ribosomal DNA (18S, 28S, ITS) and mitochondrial DNA (COI). Generally, the 18S and 28S markers are recommended for taxonomic studies at family and generic level, while the internal transcribed spacer (ITS) is useful at the species level resolution. The mitochondrial cytochrome C oxidase subunit I gene (cox1 or COI) was proposed as a universal marker for species identification and is used as a “DNA barcode” tagging any taxon in the animal kingdom. For the diverse monogenean family Dactylogyridae, the D1-D3 region of LSU (28S) is currently most frequently used molecular marker. Also, the 18S small subunit rRNA gene (SSU) in combination with ITS1 is commonly used in discriminating of dactylogyrids and their phylogenetics. Using mitochondrial COI sequences as a diagnostic molecular marker for dactylogyridean species identification is more difficult, because higher nucleotide diversity at species level has been detected and the universal barcoding primers are unavailable. In our study, we used 18S in combination with ITS1 (18S-ITS1) and partial 28S rDNA for species identification, studying the genetic relationships and evaluation of taxonomic position of three genera of dactylogyrids (Annulotrema, Afrocleidodiscus and Characidotrema) parasitizing African tetras (Characiformes: Alestidae) and six genera of dactylogyrids (Bagrobdella, Protoancylodiscoides, Synodentella, Schilbetrema, Schilbetrematoides and Quadriacanthus) of African freshwater catfishes (Siluriformes: Bagridae, Clariidae, Claroteidae, Malapteruridae, Mochokidae and Schilbeidae). All obtained sequences represent first molecular data for species studied (previously described and/or new). For both markers, no intraspecies genetic variability was detected. The highest values of genetic distances were observed for ITS1 fragment, the lowest for 18S rDNA (when we analysed these fragments separately). The partial 28S rDNA sequences were used for phylogenetic analyses in order to evaluate the position of these genera studied within other representatives of African dactylogyrids. The maximum likelihood (ML) and Bayesian analyses (BI) pointed to the monophyletic nature of all genera studied, except Schilbetrema spp. which seems to be paraphyletic. Our results (novel data for nine genera of dactylogyrids) broadening our knowledge on the diversity and evolution of gill monogeneans and may contribute to solving systematic and phylogenetic questions in this field.

Acknowledgement This research was supported by the Czech Science Foundation (project No. P505/12/G112).

The occurrence of the non-native tapeworm Khawia japonensis (Cestoda) in cultured common carp in the Czech Republic confirms its recent expansion in Europe Tomáš Scholz1*, Daniel Barčák2, Mikuláš Oros2 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Institute of Parasitology, Slovak Academy of Sciences, Hlinkova, Košice, Slovakia *[email protected]

Invasive parasites represent a serious problem due to their capacity to threaten local populations of native (often endemic) hosts, and fishes in breeding facilities. Tapeworms (Cestoda) are parasitic flatworms extremely adapted to parasitism (they lack any gut and circulatory system) and some of them have colonised new geographical regions as a result of unintentional transfer of hosts infected with these parasites. The highest number of invasive parasites within this host-parasite system is among tapeworms parasitising common carp (Cyprinus carpio L.), which has also been introduced globally. In the present account, we report another record of the Asian non-native fish tapeworm Khawia japonensis (Yamaguti, 1934) (Cestoda: Caryophyllidea) from common carp in Europe. Previous records of this cestode from Italy (Po River basin) and Slovakia (Danube River basin) and its present finding in the Czech Republic (Elbe River basin) confirms recent expansion of the parasite in Europe. Potential impacts of this non-native parasite on common carp in commercial breeding fisheries should be carefully studied (Scholz et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Scholz T., Barčák D., Oros M. (2018) The occurrence of the non-native tapeworm Khawia japonensis (Yamaguti, 1934) (Cestoda) in cultured common carp in the Czech Republic confirms its recent expansion in Europe. Bioinvasions Records 7: 303–308.

Redescription of Sciadocephalus megalodiscus Diesing, 1850, an unusual fish tapeworm (Cestoda: Proteocephalidae) Tomáš Scholz1*, Alan de Chambrier2 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Department of Invertebrates, Natural History Museum, Geneva, Switzerland *[email protected]

The tapeworm Sciadocephalus megalodiscus Diesing, 1850 (Cestoda: Proteocephalidae) is redescribed based on newly collected specimens parasitising tucunare peacock bass, Cichla monoculus Agassiz, 1831 (Perciformes: Cichlidae), in the Peruvian Amazon. Even though this cestode was redescribed two decades ago, that redescription did not report some of the unique features of this species. The most unusual characteristics of the species are: (i) peculiar formation of the uterus, with fast formation of numerous, tightly packed diverticula protruding ventrally and dorsally, with simultaneous disintegration of the ovary and vitelline follicles in the first pregravid proglottids; (ii) inverted umbrella-shaped scolex with a well-developed apical sucker; (iii) a large-sized, follicular (grape cluster-like) ovary, which occupies most of the central (median) third of proglottids, with the ovarian isthmus situated almost equatorially; (iv) regular alternation of genital pores; (v) a well-developed internal seminal vesicle; and (vi) a small- sized strobila (shorter than 6 mm) consisting of few proglottids (15–20). Preliminary molecular data reveal S. megalodiscus to be most closely related to Cichlidocestus gillesi, the type species of the recently erected Cichlidocestus de Chambrier, Pinacho-Pinacho, Hernández-Orts et Scholz, 2017. Species of both genera parasitize Neotropical cichlids and are unique among all proteocephalids in the shape and position of the ovary. They also share other morphological characteristics unusual among other proteocephalids and thus these two genera can be considered good candidates to be placed in a new, higher-level taxon such as separate subfamily or even family when a new, more natural classification of the Proteocephalidae is proposed (Scholz and de Chambrier, 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Scholz T., de Chambrier A. (2018) Redescription of Sciadocephalus megalodiscus Diesing, 1850, an unusual fish tapeworm (Cestoda: Proteocephalidae). Journal of Parasitology 104(5) (in press).

A digest of fish tapeworms Tomáš Scholz1*, Roman Kuchta1 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

Members of the following seven of a total of 19 cestode orders mature in rayfinned fishes (Actinopterygii): Amphilinidea (mainly in acipenseriforms and osteoglossiforms; 8 species in 6 genera), Bothriocephalidea (in several freshwater and marine fish groups; 129/47), Caryophyllidea (mainly in cyprinids and suckers, also in some catfishes; 117/42), Haplobothriidea (exclusively in bowfin; 2/1), Nippotaeniidea (in osmeriforms and perciforms; 6/1), freshwater Onchoproteocephalidea (mainly in catfishes 194/55), and Spathebothriidea (in several freshwater and marine fish groups; 6/5). Updated information on species diversity, host associations, interrelations and geographical distribution is provided for every group. The existing phylogenetic hypotheses suggest that tapeworms colonized rayfinned fishes several times and form several independent lineages. From a total of 461 fish tapeworms only 92 species are exclusively marine. So, freshwater species dominate the assemblage. No general patterns in host use can be observed at the level of fish definitive hosts because cestodes of fishes occur in not closely related host groups. Nevertheless, only three fish orders host almost three fourth of all tapeworms of fishes, namely Siluriformes (36 % of all cestode species), Cypriniformes (22 %) and Perciformes (16 %). Nearly two thirds (61 %) of fish tapeworms have a strict (oioxenous) specificity and one third (33 %) is stenoxenous. The highest proportion (8 %) of euryxenous species is among the bothriocephalideans, including one of the most opportunistic fish helminth, the invasive Asian fish tapeworm (Schyzocotyle acheilognathi), which has been reported from more than 200 fish species and axolotl, snakes and birds. Tapeworms of fishes are more diverse in the temperate zones of the northern hemisphere (Nearctic and Palearctic regions with 26 % and 23 % of all species) than in the tropics, with the exception of onchoproteocephalideans in South America (together with a very few other tapeworms represent 29 % of all species). However, this may reflect a lower sampling effort in tropical regions and the southern hemisphere. Caryophyllideans dominate in the Australasian (58 % of all species, but only 12 species found) and Nearctic (56 %; total number = 102) realms, and together with oncoproteocephalideans in the Ethiopian (both 41 %; n = 39), Palearctic (both 37 %; n = 90) realms, and with bothriocephalideans in the Oriental realm (34 %; n = 36). The Neotropical region is dominated by onchoproteocephalideans (in fact almost exclusively members of the family Proteocephalidae; n = 112), which represent as many as 95 % of all species reported. The major part of the undescribed diversity of fish tapeworms regarding the total number of species can be expected in the latter region, but this concerns only a single cestode family, Proteocephalidae (order Onchoproteocephalidea).

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112.

Reference 1. Scholz T., Kuchta R. (2017) A digest of fish tapeworms. Vie et Milieu 67: 43–58.

Fig 1. Zoogeography of seven orders of tapeworms parasitizing fishes as adults (some of the 359 species of freshwater fish tapeworms occur in more than one zoogeographical realm).

Fig 2. Host specificity and host spectrum of fish tapeworms with more detailed data on the three largest orders (Bothriocephalidea, Caryophyllidea and Onchoproteocephalidea – family Poteocephalidae). Graphs below: Host spectrum of all fish tapeworms by frequencies of 36 fish orders. Fish icons showing the most common fish orders. Host spectrum of the bothriocephalideans by frequencies of 97 reported fish families. Host spectrum of the caryophyllideans by frequencies of 12 reported fish families. Host spectrum of onchoproteocephalideans (proteocephalids) by frequencies of 47 reported fish families.

The first record of the invasive Asian fish tapeworm (Schyzocotyle acheilognathi) from an endemic cichlid fish in Madagascar Tomáš Scholz1*, Andrea Šimková2, Jeanne R. Razanabolana3, Roman Kuchta1 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 3Department of Animal Biology, Faculty of Science, University of Antananarivo, Antananarivo, Madagascar *[email protected]

The Asian fish tapeworm, Schyzocotyle acheilognathi (Yamaguti, 1934) (Cestoda: Bothriocephalidea), is an invasive parasite of freshwater fishes that have been reported from more than 200 freshwater fish worldwide. It was originally described from a small cyprinid, Acheilognathus rhombeus, in Japan but then has spread, usually with carp, minnows or guppies, to all continents including isolated islands such as Hawaii, Puerto Rico, Cuba or Sri Lanka. In the present account, we report the first case of the infection of a native cichlid fish, Ptychochromis cf. inornatus (Perciformes: Cichlidae), endemic to Madagascar, with S. acheilognathi. The way of introduction of this parasite to the island, which is one of the world’s biodiversity hotspots, is briefly discussed (Scholz et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Scholz T., Šimková A., Rasamy Razanabolana J., Kuchta R. (2018) The first record of the invasive Asian fish tapeworm (Schyzocotyle acheilognathi) from an endemic cichlid fish in Madagascar. Helminthologia 55: 84– 87.

An annotated list and molecular data on larvae of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) from freshwater fishes in Africa Tomáš Scholz1*, Sareh Tavakol2, Lucie Uhrová1, Jan Brabec1, Iva Přikrylová2,3,4, Šárka Mašová3, Andrea Šimková3, Ali Halajian2, Wilmien J. Luus-Powell2 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences & University of South Bohemia, České Budějovice, Czech Republic; 2Department of Biodiversity, University of Limpopo, Sovenga, South Africa; 3Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 4Water Research Group, Unit for Environmental Sciences and Management, North-West University, Potchefstroom, South Africa *[email protected]

An annotated list of larvae (metacestodes) of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) from freshwater fishes in Africa is provided with numerous new host and geographical records. Newly collected materials from Burundi, Democratic Republic of the Congo, Kenya, Madagascar, Namibia, Senegal, South Africa, Sudan and Zimbabwe practically double the total number of species reported from African fish so far. We confirm the occurrence of 16 species (five unidentified to the species level and most likely representing new taxa) belonging to the genera Amirthalingamia Bray, 1974 (1 species), Cyclustera Fuhrmann, 1901 (2), Dendrouterina Fuhrmann, 1912 (1), Neogryporhynchus Baer & Bona, 1960 (1), Paradilepis Hsü, 1935 (4), Parvitaenia Burt, 1940 (5), and Valipora Linton, 1927 (2). Besides metacestodes of four unidentified species of Paradilepis and Parvitaenia, Paradilepis maleki Khalil, 1961 is reported from fish for the first time. Rostellar hooks of all species are illustrated and their measurements are provided. The molecular phylogenetic analysis based on partial lsrDNA sequences offers the first insight into the internal phylogenetic relationships within the family. Together with the morphological observations, the present study provides a taxonomic baseline for future studies on this largely neglected, but widely distributed and relatively frequent, group of parasites of African fishes, including economically important cichlids like tilapias and cyprinids (Scholz et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Scholz T., Tavakol S., Uhrová L., Brabec J., Přikrylová I., Mašová Š., Šimková A., Halajian A., Luus-Powell, W.J. (2018) An annotated list and molecular data on larvae of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) from freshwater fishes in Africa. Systematic Parasitology 95: 567–590.

Guide to the Parasites of African Freshwater Fishes: Diversity, Ecology and Research Methods Tomáš Scholz1*, Maarten P.M. Vanhove2,3,4,5,6, Nico J. Smit1, Zuzana Jayasundera2, Milan Gelnar2 (Editors) 1Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic; 2Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 3Operational Directorate Taxonomy and Phylogeny, Royal Belgian Institute of Natural Sciences, Brussels, Belgium; 4Laboratory of Biodiversity and Evolutionary Genomics, Department of Biology, University of Leuven, Leuven, Belgium; 5Centre for Environmental Sciences, Research Group Zoology: Biodiversity & Toxicology, Universiteit Hasselt, Diepenbeek, Belgium; 6 Zoology Unit, Finnish Museum of Natural History, University of Helsinki, Helsinki, Finland *[email protected]

The rivers and lakes of Africa contain almost 25 % of the world’s 13,000 freshwater fish species and are second only to South America in species richness. These fish are parasitised by a wide range of organisms that can be detrimental to both farmed and wild fishes with consequent effects on economic development, and often on human health. Knowledge of these parasites in African freshwater fishes is limited and this book is intended to promote and advance understanding of African fish parasites by providing information on the best techniques for investigating fish and their parasites and keys to parasite identification. The first comprehensive list of all known freshwater fish parasites in Africa is presented here, with information on their known hosts and distribution, keys to all genera and representative illustrations for every genus. This information should facilitate and stimulate the development of fish parasitology on the African continent which has great potential for aquaculture and fishery development (Scholz et al., 2018).

Dedication This book is dedicated to the memory of Jo Van As, University of the Free State, Bloemfontein, South Africa, for his extraordinary contribution to fish parasitology in Africa.

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Scholz T., Vanhove M.P.M., Smit N., Jayasundera Z. and Gelnar M. (Eds.) (2018) Guide to the Parasites of African Freshwater Fishes: Diversity, Ecology and Research Methods. ABC Taxa 18: 1–417.

Diversity of monogeneans of freshwater fish and coevolution in monogenean-fish systems Andrea Šimková1* 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

Gill monogeneans exhibiting the high host specificity to their fish hosts are powerful models to investigate host- parasite coevolution. However, the knowledge on monogenean diversity, crucial for ecological and evolutionary studies, are still currently underestimated in many freshwater fish groups. During 2018, I focused on data analyses using three models of monogeneans-freshwater fish in order to summarize the knowledge on monogenean diversity in the host-parasite systems selected, to test phylogenetic hypotheses and to perform the cophylogenetic analyses in order to infer the coevolutionary processes in the following host-parasite systems. The first phylogenetic study was performed to investigate the phylogeny of Dactylogyrus parasitizing North African cyprinids belonging to two phylogenetic lineages (1) Carasobarbus fritschii and Pterocapoeta maroccana - representatives of hexaploid Torini, and (2) Luciobarbus representatives of tetraploid Barbini. From 13 Dactylogyrus species reported in 10 Moroccan cyprinid species, 7 were identified as new species. 11 from 13 Dactylogyrus species were reported in a single host species. Dactylogyrus of Torini formed the monophyletic group well separated from monophyletic group of Dactylogyrus parasitizing Luciobarbus. D. marocanus was a single species switching four cyprinid species belonging to Torini and Barbini and this species was not clustered with host-specific Dactylogyrus of Torini or host-specific Dactylogyrus of Barbini. Both cophylogenetic methods revealed the significant cophylogenetic signal. Using distance based methods, the majority of host-parasite associations significantly contributed to the whole cophylogenectic structure. The cospeciation was evidenced for Dactylogyrus-cyprinids considering host tribus level. Nevertheless, host switch represented the most common evolutionary event of Dactylogyrus speciation in morphologically similar and rapidly evolving Luciobarbus. The next phylogenetic study was performed to investigate the phylogenetic position of Anacanthorus spp. infesting serrasalmids from two Brazilian river basins, within Dactylogyridae. Anacanthorus is the most specious genus parasitizing Characiformes, especially Bryconidae, Characidae and Serrasalmidae. In this study, a total of 18 Anacathorus species including 8 species recognized as new for science were reported from 11 serrasalmid host species were investigated. Following the previous study based on morphological data, we hypothesized and consequently supported the monophyly of Anacanthorus. We showed that the representative of other monogenean genus i.e. Mymarothecium is not sister species to Anacanthorus group. The phylogenetic analyses indicated that the relationships among Anacanthorus species reflect the relationships between the lineages of the serrasalmid hosts: the first subgroup includes a species specific to hosts of the genus Piaractus, member of the “pacus” lineage, the second subgroup includes a species from the “Myleus-like pacus” lineage and the third subgroup includes species from the lineage of the “true piranhas”. The last phylogenetic study was focused to investigate the phylogenetic position of monogeneans specific to Malagasy cichlids. The aim of this study was to use the host specific parasites to infer biogeographical scenario of worldwide distribution of their cichlid hosts. We showed that monogeneans specific do Malagasy cichlids do not form a monophyletic group which is in concordance with non-monophyletic origin of Malagasy cichlids. However, our analyses revealed the importance of morphological re-investigation of Malagasy monogeneans prior to finalize the phylogenetic study. Currently new material has been collected be foreign collaborators, morphological and molecular analyses will be performed. Finally, the field trip in Peru was performed to investigate the diversity and phylogeny of monogeneans specific to cichlids from Amazon River. We hypothesized that the phylogeny of host specific gill monogeneans follow the phylogeny of diverse cichlid lineages. The analyses of the morphological diversity and molecular data of monogeneans are currently under process.

Acknowledgement This study was partially supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112. I acknowledge all colleagues and students participating on field trips and data collection.

New data on cyst ultrastructure of the amoebozoan species Flamella arnhemensis Tomáš Tyml1*, Iva Dyková1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic *[email protected]

The genus Flamella Schaeffer, 1926 contains 12 nominal species. In four of those (F. citrensis, F. lacustris, F. magnifica and F. tiara), data on cyst formation are completely missing, while the descriptions of cysts of another two species (F. aegyptia and F. piscinae) are very short and simple. More or less detailed and better-documented descriptions of cyst structure have only been provided for F. arnhemensis, F. balnearia and F. fluviatilis by Kudryavtsev et al. [2]. Recently, some data on the cyst structure have also been published for F. daurica [1], F. pleistocenica and F. beringiana [3], but the quality of documentation on the cyst ultrastructure of the latter three species is inferior to that by Kudryavtsev et al. [2], making comparison of these data hardly possible. Our study of a novel Flamella strain isolated from fresh horse droppings allowed us to supplement the data on the ultrastructure of the cyst wall of Flamella as published by Kudryavtsev et al. [2]. By light microscopy, our newly isolated strain displayed all the characteristics of the diagnosis of Flamella Schaeffer, 1926 emended by Kudryavtsev et al. [2]. The sequence of the 18S rRNA gene (deposited into the DDBJ/EMBL/NCBI databases under Acc. No LC371660) is identical with 18S rRNA gene sequence of Flamella arnhemensis, strain CCAP 1556/1. Considering the current state of knowledge of the phylogeny within the genus Flamella [1] and the other pairwise distances to Flamella sequences, our newly isolated strain (denominated EQIN) was unambiguously identified with Flamella arnhemensis Kudryavtsev, Wylezich, Schlegel, Walochnik & Michel, 2009. Our comprehensive study of developing and mature cysts of EQIN strain resulted in an ultrastructural specification of the three-layer cyst wall consisting of an innermost, sharply delimited layer 150–300 nm thick, closely apposed to the encysted trophozoite; a middle layer 200–400 nm thick, sharply delimited only against the innermost layer; and an outer layer consisting of loosely arranged fibrils and an ectocyst outer sheet. At low magnifications, the middle layer had an electron dense granular appearance (Fig C). Higher magnifications, however, revealed that the “granular” middle layer was, in fact, composed of electron-dense material arranged in a reticular network. The pattern of this network was difficult to interpret in cross-sectioned cysts (Figs D, E), however, it could be better understood if artificially detached from the innermost layer (Fig F). The observation of excystment (Fig G) supported these findings. The data are consistent with those given for the CCAP 1556/1 strain of F. arnhemensis by Kudryavtsev et al. [2] in terms of the number of cyst wall layers, however, the relatively low magnification used by these authors prevented them from recognizing the true nature of the “middle granular layer”. A reliable, convincing documentation of the cyst ultrastructure is missing for most Flamella nominal species. This is due to common problems in the preparation of cyst-forming organisms for transmission electron microscopy which include poor penetration with fixatives, insufficient dehydration, incomplete impregnation with resin, and overstaining of cyst walls. Difficult as it may be, mastering the preparation techniques and a careful selection of electronograms for publication cannot be circumvented if one wants to achieve quality results.

Acknowledgement This research was supported by the Czech Science Foundation (grant 505/12/G112) and the Ministry of Education, Youth and Sports (grant LM2015062 Czech-BioImaging).

References 1. Glotova A., Smirnov A.V. (2017) Description of Flamella daurica n. sp., with notes on the phylogeny of the genus Flamella and related taxa. European Journal of Protistology 58: 164–174. 2. Kudryavtsev A., Wylezich C., Schlegel M., Walochnik J., Michel R. (2009) Ultrastructure, SSU rRNA gene sequences and phylogenetic relationships of Flamella Schaeffer, 1926 (Amoebozoa), with description of three new species. Protist 160: 21–40. 3. Shmakova L., Bondarenko N., Smirnov A. V. 2016. Viable species of Flamella (Amoebozoa: Variosea) isolated from ancient Arctic permafrost sediments. Protist 167: 13–30.

Fig 1. A. Trophozoites of Flamella arnhemensis as seen in a light microscope; B. Cysts of F. arnhemensis as seen in the light microscope; C. Overview of a cyst observed in a transmission electron microscope. The cyst wall consists of an outer layer (ol) composed of loosely arranged fibrils and an ectocyst outer sheet, an electron dense middle layer (rl) of granular appearance at this low magnification, and a homogeneous inner most layer (il); D, E. Cross sections through the cyst wall showing part of an encysted trophozoite (et), inner layer (il), and reticular layer (rl); F. Mesh- like structure of the reticular layer partly detached from the inner layer of a cyst wall; G. Overview of trophozoite excystment showing the three-layer cyst wall and trophozoite emerging through the ostiole.

Exploring mitogenomics for the phylogeny of African monogeneans (Gyrodactylidae and Dactylogyridae) Maarten P.M. Vanhove1,2,3,4*, Andrew Briscoe5, Michiel W.P. Jorissen2,6, Tim J. Littlewood5, Tine Huyse4,6 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic 2Hasselt University, Centre for Environmental Sciences, Research Group Zoology: Biodiversity & Toxicology, Diepenbeek, Belgium 3Zoology Unit, Finnish Museum of Natural History, University of Helsinki, Helsinki, Finland 4Laboratory of Biodiversity and Evolutionary Genomics, Department of Biology, University of Leuven, Leuven, Belgium 5Department of Life Sciences, Natural History Museum, London, United Kingdom 6Invertebrate Section, Biology Department, Royal Museum for Central Africa, Tervuren, Belgium *[email protected]

Gyrodactylidae and Dactylogyridae are probably the most species-rich monogenean families, with representatives that may be harmful fish pathogens, especially in intensive aquaculture or after anthropogenic co-introduction of host and parasite. In Africa, cichlid and clariid fishes are quite well-studied for monogeneans, among which the most important aquaculture fishes on the continent, many of which have been widely introduced outside Africa [1]. A limitation, however, is the small number of available molecular markers for these worms. Several questions on their evolution, such as the phylogenetic position of Macrogyrodactylus and the highly species-rich Gyrodactylus, remain unresolved with available markers. The need for additional and higher-resolution markers is especially clear when investigating processes on a recent timescale, such as phylogeography, population genetics, and barcoding. A small set of established mitochondrial gene fragments constitutes the most variable markers available. They also served in flatworm systematics on a higher, macro-evolutionary scale. We initiated a mitogenomic approach to African monogenean evolution, starting with species collected from the Upper Congo Basin [2]. Based on Illumina technology, mitochondrial genomes were assembled and annotated for the cichlid parasites Gyrodactylus nyanzae, Cichlidogyrus halli, Cichlidogyrus mbirizei (near-complete mitogenome) and the catfish parasite Macrogyrodactylus karibae (near-complete mitogenome) [3]. We report a new start codon (TTG) for Gyrodactylus and for Dactylogyridae, and a new, incomplete stop codon (TA) for Dactylogyridae. A mitochondrial phylogeny based on a nearly 12,500 bp alignment confirms the basal position of the Neotropical oviparous Aglaiogyrodactylus (and not of the African Macrogyrodactylus) with respect to other gyrodactylids, and confirms the paraphyly of Gyrodactylus. Within dactylogyrids, additional taxon sampling is needed to increase phylogenetic resolution, although gene order suggests Cichlidogyrus to be closely related to marine ancyrocephalines. Gene order is identical for protein-coding genes and differs between the African representatives of these families only in a tRNA gene transposition. These first mitogenomic data on African monogeneans underscore the potential of mitochondrial genes and gene order in flatworm phylogenetics.

Future plans Further exploration of mitogenomics and manuscript writing on other aspects of the (molecular) evolution and ecology of monogenean fish parasites of Lake Tanganyika and other parts of the Congo Basin, plus finalisation of several taxonomic manuscripts.

Acknowledgement This study was supported by ECIP (CSF project no. P505/12/G112), BELSPO BRAIN-be Pioneer Project BR/132/PI/TILAPIA, VLIR-UOS South Initiative ZRDC2014MP084, SYNTHESYS and FWO-Vlaanderen.

References 1. Vanhove M.P.M., Hablützel P.I., Pariselle A., Šimková A., Huyse T., Raeymaekers J.A.M. (2016) Cichlids: a host of opportunities for evolutionary parasitology. Trends in Parasitology 32(10): 820–832. 2. Jorissen M.W.P., Pariselle A., Huyse T., Vreven E.J., Snoeks J., Volckaert F.A.M., Chocha Manda A., Kapepula Kasembele G., Artois T., Vanhove M.P.M. (2018) Diversity, endemicity and host-specificity of monogenean gill parasites (Platyhelminthes) of cichlids in the Bangweulu-Mweru ecoregion. Journal of Helminthology 92(4): 417–437. 3. Vanhove M.P.M., Briscoe A.G., Jorissen M.W.P., Littlewood D.T.J., Huyse T. (2018) The first next-generation sequencing approach to the mitochondrial phylogeny of African monogenean parasites (Platyhelminthes: Gyrodactylidae and Dactylogyridae). BMC Genomics 19: 520.

Eudiplozoon nipponicum (Polyopisthocotylea, Diplozoidae): third monogenean genome at the reach of the hand? Jiří Vorel1*, Marie Jankůjová2, Jan Oppelt2, Filip Pardy3, Pavel Roudnický1, Jana Ilgová1, Milan Gelnar1, Martin Kašný1 1Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czech Republic; 3Central European Institute of Technology, Masaryk University, Brno, Czech Republic *[email protected]

Actual information related to the genomes and other “omic” data of monogeneans is limited on two genomes (Gyrodactylus salaris and Protopolystoma xenopodis), one transcriptome (Neobenedenia melleni) and a few described mitochondrial genomes. In 2013 we started with sequencing of E. nipponicum nuclear genome. For the first genome draft, 164,773,962 raw reads were generated by 454/Roche (Junior sequencing platform) and Illumina (MiSeq and HiSeq sequencing platforms) instruments, bioinformatically processed and assembled into 524,914 scaffolds with total length 1.1 Gb, which is not entirely corresponding with estimated genome size – 1.5 Gb. For the purpose to improve this first genome draft, we have additionally included long reads generated by Oxford Nanopore (MinION sequencing platform). Using three flow-cells 2,755,771 reads (8.0 Gb) were produced with an average read length 2,901 bp and good quality. The complex robust hybrid assembly of these short and long reads is currently under bioinformatic processing. Althought the analysis is not complete, based on the first genome draft data, we were able identified and additionally assembled (NOVOplasty and MaSuRCA assemblers) complete sequence of E. nipponicum mitochondrial genome. With the size 16,727 bp is E. nipponicum mitochondrial genome the longest mitogenome within the monogeneans. We performed the annotation, carried on by MITOS tool and 36 genes with features common in other platyhelminths were characterized, e.g. long non-coding regions, same direction of PCGs genes, overlap regions of nad genes and asymmetric distribution of nucleotide composition.

Acknowledgement This research was supported by Czech Science Foundation (GBP505/12/G112, P506/12/1258) and grants of the Masaryk University (MUNI/A/1362/2016, MUNI/A/0816/2017).

Round goby (Neogobius melanostomus) in the food chain of two Czech rivers. Lucie Všetičková1*, Libor Mikl1, Zdeněk Adámek1, Mojmír Vašek1, Helena Švecová2, Jitka Kolářová2, Pavel Jurajda1 1Institute of Vertebrate Biology, Czech Academy of Sciences, Brno, Czech Republic; 2Faculty of Fisheries and Protection of Waters, Research Institute of Fish Culture and Hydrobiology in Vodňany, University of South Bohemia in České Budějovice, Czech Republic *[email protected]

Round goby (Neogobius melanostomus), an invasive Ponto-Caspian species, was first recorded in the Rivers Dyje and Morava in 2008. The species is fast growing, a batch spawner and shows tolerance to a wide range of abiotic factors, greatly increasing its invasive potential. Following their introduction into the Laurentian Great Lakes in the 1990s, a number of negative impacts were confirmed on indigenous fish abundance (Jude et al. 2005). Additionally, the species appeared to play a negative role in food chain mercury transmission (Hogan 2007, Kornis et al. 2012) due to a dietary preference for dreissenid mussels (Dreissena polymorpha). As round gobies continue to spread, it becomes increasingly important to assess the level of dietary and/or habitat competition with indigenous fish species and to assess whether the negative impacts observed in the USA can be generalised to other areas. The aim of this study was to assess round goby impact in two Central European rivers, the Dyje and Elbe. In the Elbe, round gobies mainly consumed a small non-indigenous crustacean (Dikerogammarus vilosus; frequency of occurrence 63 %), chironomid larvae (33 %) and trichopteran larvae (27 %). Dreissenid mussels and fish were taken less frequently (15 % and 2%, respectively). All fish in the diet were from the Gobiidae family. In the Dyje, gobies fed mainly on chironomid larvae (66 %), trichopteran larvae (63 %) and ephemeropteran larvae (53 %), with dreissenid mussels and fish (Gobiidae) again taken less often (22 % and 3 %, respectively). At the same time, round gobies became a common prey item of predatory fish in the Dyje, representing around 20–30 % of pike (Esox lucius) and burbot (Lota lota) diet, and 10-20 % of prey taken by perch (Perca fluviatilis), wells (Silurus glanis) and Volga zander (Sander volgense). Mercury content in round goby muscle varied between 0.08 and 0.1 ppm (similar to levels in gudgeon Gobio gobio), while levels in dreissenid mussels were negligible at between 0.01 and 0.02 ppm. In conclusion, round goby diet consisted mainly of benthic invertebrate, with dreissenid mussels taken only occasionally. Fish were taken only rarely (˂ 5 %) and were represented by Gobiidae sp. only. Mercury content was the same as that in the other benthic fish species. Hence, round gobies appear to be having no negative impact as regards trophic transfer of mercury or as a predator of indigenous fish species, though they have become a prey species of indigenous fish predators.

Acknowledgement This study was supported by the European Centre of Ichthyoparasitology Project No. ECIP P/505/12/G112.

References 1. Hogan L.S., Marschall E., Folt C., Stein R.A. (2007) How non-native species in Lake Erie influence trophic transfer of mercury and lead to top predators. Journal of Great Lakes Research 33: 46–61. 2. Jude D.J., Janssen J., Crawford G. (1995) Ecology, distribution, and impact of the newly introduced round & tubenose gobies on the biota of the St. Clair & Detroit Rivers. In: Munawar M., Edsall T., Leach J. (eds.): The Lake Huron ecosystem: Ecology, Fisheries and Management. SPB Academic Publishing, Amsterdam, pp 447– 460. 3. Kornis M.S., Mercado-Silva N., Vander Zanden M.J. (2012) Twenty years of invasion: a review of round goby Neogobius melanostomus biology, spread and ecological implications. Journal of Fish Biology 8: 235–285.

A new species of Aphanoblastella Kritsky, Mendoza-Franco and Scholz, 2000 (Monogenea, Dactylogyridae) parasitic on heptapterid catfish (Siluriformes) in the Neotropical Region Fabio Hideki Yamada1*, Aline A. Acosta2,3, Priscilla de O.F. Yamada2, Tomáš Scholz3, Reinaldo J. da Silva2 1Regional University of Cariri (URCA), Crato, Ceará State, Brazil; 2Institute of Biosciences, Department of Parasitology, São Paulo State University (UNESP), Botucatu, São Paulo, Brazil; 3Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

Aphanoblastella magna n. sp. is described from the gills of the heptapterid catfish Pimelodella avanhandavae Eigenmann, 1917 in southeastern Brazil (Paraná River Basin). This new species most closely resembles Aphanoblastella chagresii Mendoza- Franco, Aguirre-Macedo and Vidal-Martínez, 2007 described from a congeneric fish host in Panama, but can be distinguished by the shape of the accessory piece and shape and size of the ventral and dorsal bars. From the other species of the genus, A. magna n. sp. differs mainly in the shape of the male copulatory organ (MCO), which is sinuous, versus spirally coiled in other species of the genus, except for A. travassosi (Price, 1938), and by the accessory piece which resembles a ‘shoehorn’. Aphanoblastella magna n. sp. is the seventh species of the genus and the first dactylogyrid described from P. avanhandavae. A partial 28S rDNA gene sequence of the new species is also provided; it forms a monophyletic clade with two congeners for which molecular data are available (Yamada et al., 2018).

Acknowledgement This study was supported by the Czech Science Foundation (project No. P505/12/G112).

Reference 1. Yamada F.H., Acosta A.A., Yamada P.O.F., Scholz T., Silva R.J. (2018) A new species of Aphanoblastella Kritsky, Mendoza-Franco and Scholz 2000 (Monogenea, Dactylogyridae) parasitic on heptapterid catfish (Siluriformes) in the Neotropical Region. Acta Parasitologica 63(4): 772–780.

Study of the tegument, glands and the excretory system of plerocercoids of Dibothriocephalus latus (Cestoda: Diphyllobothriidea) Aneta Yoneva1,2*, Tomáš Scholz2, Roman Kuchta2 1Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, Sofia, Bulgaria; 2Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic *[email protected]

Study of the surface, glands and the protonephridial system of plerocercoids of diphyllobothriidean cestode Dibothriocephalus latus was made using transmission electron microscopy (TEM). Electron microscopy revealed two different types of microtriches on the scolex and body surface of plerocercoids of D. latus, hamulate spinitriches and capilliform filitriches. Electron-dense discoidal and spherical bodies and numerous vesicles appeared in the distal cytoplasm. Nonciliated sensory receptors have been observed between the distal cytoplasm and the underlying musculature of the plerocercoids. The plerocercoids of D. latus possess a well-developed gland complex filling the parenchyma of the whole body. Two types of glands composed of different types of secretory granules of various size and shape have been observed in the scolex and body parenchyma of D. latus plerocercoids. It is suggested that the secretions probably play a role in adhesion of the worm during its migration into the host tissue. Our study revealed that the gland cells of type two and their secretion in D. latus plerocercoids are morphologically similar to those described by Kuperman and Davydov [1] who showed only one type of glands in plerocercoids of D. latus. The fine structure of the excretory system in the plerocercoids of D. latus shows an organization similar to that observed in the adults of that species. It comprises two main components, the flame cells which are the basic units of the protonephridial system of invertebrates and the excretory ducts. By TEM, flame cells exhibit a large nucleus surrounded by scant cytoplasm containing mitochondria, ribosomes and vesicles, and classical cilia with 9+2 axonemes. Although the function of the flame cells has not yet precisely defined, there is a consensus that they perform excretion/secretion activities. The present study has revealed marked differences in the fine structure of microtriches and gland cells between the plerocercoid and adult stages of D. latus [2].

Future plans Submission of the manuscript concerning the data presented in this abstract. Extending this survey to obtain new information on the surface structures, glands, sensory organs and the excretory system in plerocercoid and adult stages of species of Spirometra as a basis for subsequent comparative analyses.

Acknowledgement This study was supported by ECIP (European Centre of Ichthyoparasitology); Centre of excellence program of the Czech Science Foundation; project No. P505/12/G112. We acknowledge the Laboratory of Electron Microscopy, Institute of Parasitology, Biology Centre of CAS, supported by the MEYS CR (LM2015062 Czech-Bioimaging) for technical assistance.

References 1. Kuperman B.I., Davydov V.G. (1981) The fine structure of glands in oncospheres, procercoids and plerocercoids of Pseudophyllidea (Cestoidea) International Journal for Parasitology 12: 135–144. 2. Yoneva A., Scholz T., Kuchta R. (2018) Comparative morphology of surface ultrastructure of diphyllobothriidean tapeworms (Cestoda: Diphyllobothriidea) Invertebrate Biology 137: 38–48.

List of all publications dedicated to ECIP Actual or past members of ECIP are presented in bold. List is divided into parts according to year of publication.

2018 Acosta A.A., Scholz T., Blasco-Costa I., Alves P.V., da Silva R.J. (2018) A new genus and two species of dactylogyrid monogeneans from gills of Neotropical catfishes (Siluriformes: Dorididae and Loricariidae). Parasitology International 67: 4–12. [IF2017 = 1.886] Alves P.V., de Chambrier A., Luque J.L., Scholz T. (2018) Towards a robust systematic baseline of Neotropical fish tapeworms (Cestoda: Proteocephalidae): amended diagnoses of two genera from the redtail fish Phractocephalus hemioliopterus. Zootaxa 4370(4): 363–380. [IF2017 = 0.931] Bartáková V., Bryja, J., Reichard M. (2018) Fine-scale genetic structure of the European bitterling at the intersection of three major European watersheds. BMC Evolutionary Biology 18: 105. [IF2017 = 2.861] Bartošová-Sojková P., Lövy A., Reed C., Lisnerová M., Tomková T., Holzer A., Fiala I. (2018) Life in a rock pool: radiation and population genetics of myxozoan parasites in hosts inhabiting restricted spaces. PLoS ONE 13: e0194042. [IF2017 = 2.859] Blažek R., Polačik M., Smith, C., Honza, M., Meyer, A., Reichard M. (2018) Success of cuckoo catfish brood parasitism reflects coevolutionary history and individual experience of their cichlid hosts. Science Advances 4: eaar4380. [IF2017 = 11.039] Cantatore D.M.P., Irigoitia M.M., Holzer A.S., Bartošová-Sojková P., Pecková H., Fiala I., Timi J.T. (2018) The description of two new species of Chloromyxum Mingazzini, 1890 from skates in the Argentine Sea reveals that a limited geographic host distribution causes phylogenetic lineage separation of myxozoans in Chondrichthyes. Parasite 25: 47. [IF2017 = 2.069] García D., Loureiro M., Machín E., Reichard M. (2018) Phenology of three coexisting annual fish species: seasonal patterns in hatching dates. Hydrobiologia 809: 323–337. [IF2017 = 2.022] Ghanmi N., González-Solís D., Gargouri L. (2018) Two new gonad-infecting of Philometra Costa, 1845 (Nematoda: Philometridae) from Trachinus spp. (Osteichthyes: Trachinidae) in the Gulf of Hammamet, Tunisia. Systematic Parasitology 95: 223–234. [IF2017 = 1.273] Hodová I., Sonnek R., Gelnar M., Valigurová A. (2018) Architecture of Paradiplozoon homoion: A diplozoid monogenean exhibiting highly-developed equipment for ectoparasitism. PLoS ONE 13(2): [IF2017 = 2.859] Holzer A.S., Bartošová-Sojková P., Born-Torrijos A., Lövy A., Hartigan A., Fiala I. (2018) The joint evolution of the Myxozoa and their alternate hosts: A cnidarian recipe for success and vast biodiversity. Molecular Ecology 27(7): 1651–1666. [IF2017 = 6.131] Janáč M., Roche R., Šlapanský L., Polačik M., Jurajda P. (2018) Long-term monitoring of native bullhead and invasive gobiids in the Danubian rip-rap zone. Hydrobiologia 807: 263–275. [IF2017 = 2.022] Janko K., Pačes J., Wilkinson-Herbots H., Costa R.J., Roslein J., Drozd P., Iakovenko I, Rídl J., Hroudová M., Kočí J., Reifová R., Šlechtová V., Choleva L. (2017) Hybrid asexuality as a primary postzygotic barrier between nascent species: On the interconnection between asexuality, hybridization and speciation. Molecular ecology 27: 248–263. [IF2017 = 5.612] Jedličková L., Dvořáková H., Dvořák J., Kašný M., Ulrychová L., Vorel J., Žárský V., Mikeš L. (2018) Cysteine peptidases of Eudiplozoon nipponicum: a broad repertoire of structurally assorted cathepsins L in contrast to the scarcity of cathepsins B in an invasive species of haematophagous monogenean of common carp. Parasites & Vectors 11: 1–17. [IF2017 = 3.086] Jirsová D., Ding X., Civáňová K., Jirounková E., Ilgová J., Koubková B., Kašný M., Gelnar M. (2018) Redescription of Paradiplozoon hemiculteri (Monogenea, Diplozoidae) from the type host Hemiculter leucisculus, with neotype designation. Parasite 25: 4. [IF2017 = 1.745] Jorissen M.W.P., Pariselle A., Huyse T., Vreven E.J., Snoeks J., Volckaert F.A.M., Chocha Manda A., Kapepula Kasembele G., Artois T., Vanhove M.P.M. (2018) Diversity and host specificity of monogenean gill parasites (Platyhelminthes) of cichlid fishes in the Bangweulu-Mweru ecoregion. Journal of Helminthology 92: 417– 437. [IF2017 = 1.262] Kičinjaová M.L., Barson M., Gelnar M., Řehulková E. (2017) Two new species of Annulotrema (Monogenea: Dactylogyridae) from Hydrocynus vittatus (Characiformes: Alestidae) in Lake Kariba, Zimbabwe. Journal of Helminthology 92(4): 467–476. [IF2017 = 1.420] Kmentová N., Van Steenberge M., Thys van den Audenaerde D., Nhiwatiwa T., Muterezi Bukinga F., Mulimbwa N'sibula T., Masilya Mulungula P., Gelnar M., Vanhove M.P.M. (2018) Co-introduction success of monogeneans infecting the fisheries target Limnothrissa miodon differs between two non-native areas: the potential of parasites as a tag for introduction pathway. Biological Invasions. DOI: 10.1007/s10530-018- 1856-3. [IF2017 = 3.128] Kmentová N., Van Steenberge M., Raeymaekers J.A.M., Koblmüller S., Hablützel P.I., Muterezi Bukinga F., Mulimbwa N'sibula T., Masilya Mulungula P., Nzigidahera B., Ntakimazi G., Gelnar M., Vanhove M.P.M. (2018) 80

Monogenean parasites of sardines in Lake Tanganyika: diversity, origin and intra-specific variability. Contributions to Zoology 87: 105–132. [IF2017 = 1.972] Koblmüller S., Zangl L., Börger C., Daill D., Vanhove M.P.M., Sturmbauer C., Sefc K.M. (2018) Only true pelagics mix: Comparative phylogeography of deepwater bathybatine cichlids from Lake Tanganyika. Hydrobiologia. DOI: 10.1007/s10750-018-3752-3. [IF2017 = 2.022] Kvach Y., Bryjová A., Sasal P., Winkler H.M. (2018) The taxonomic and phylogenetic status of digeneans from the genus Timoniella (Digenea: Cryptogonimidae) in the Black and Baltic Seas. Journal of Helminthology 92: 596–603. [IF2017 = 1.262] Kvach Y., Ondračková M., Janáč M., Jurajda P. (2018) Methodological issues affecting the study of fish parasites. III. Effect of fish preservation method. Diseases of Aquatic Organisms 127: 213–224. [IF2017 = 1.566] Kvach Y., Ondračková M., Kutsokon Y., Dzyziuk N. (2018) New record of monogenean parasites on non-indigenous fishes in the Ukrainian Danube Delta. BioInvasions Records 7: 65–72. [IF2017 = 1.242] Moravec F., Barton D.P. (2018) Capillaria appendigera n. sp. (Nematoda: Capillariidae) from the goldbanded jobfish Pristipomoides multidens (Day) (Lutjanidae) and new records of other intestinal capillariids from marine perciform fishes off Australia. Systematic Parasitology 95: 55–64. [IF2017 = 1.273] Moravec F., Barton D.P. (2018) New records of philometrids (Nematoda: Philometridae) from marine fishes off Australia, including description of four new species and erection of Digitiphilometroides gen. n. Folia Parasitologica 65: 005. [IF2017 = 1.505] Moravec F., Cutmore S.C., Yong R.Q.-Y. (2018) Redescription of Philometra pellucida (Jägerskiöld, 1893) (Nematoda: Philometridae) parasitic in the abdominal cavity of the blackspotted puffer Arothron nigropunctatus (Bloch & Schneider) (Teleostei: Tetraodontidae) of Australia and Japan. Systematic Parasitology 95: 665–671. [IF2017 = 1.273] Moravec F., Justine J.-L. (2018) Rasheedia n. nom. (Nematoda, Physalopteridae) for Bulbocephalus Rasheed, 1966 (a homonym of Bulbocephalus Watson, 1916), with description of Rasheedia heptacanthi n. sp. and R. novaecaledoniensis n. sp. from perciform fishes off New Caledonia. Parasite 25: 39. [IF2017=2.069] Moravec F., Justine J.-L. (2018) Three new species of Cucullanus (Nematoda: Cucullanidae) from marine fishes off New Caledonia, with a key to species of Cucullanus from Anguilliformes. Parasite 25: 51. [IF2017 = 2.069] Moravec F., Nagasawa K., Madinabeitia I. (2018) Redescription of Dichelyne (Cucullanellus) jialaris (Nematoda: Cucullanidae), a parasite of seabreams (Perciformes: Sparidae) in East Asia. Acta Parasitologica 63: 802– 807. [IF2017 = 1.039] Moravec F., Novacovsky G.N., Hernández-Orts J.S. (2018) Pseudodelphis eleginopsis n. sp. (Nematoda: Guyanemidae), a new tissue-dwelling parasite of the Patagonian blennie Eleginops maclovinus (Cuvier) (Perciformes: Eleginopsidae) in Argentina, with notes on related forms. Systematic Parasitology 95: 403– 414. [IF2017 = 1.273] Moravec F., Yooyen T., Sanprick A. (2018) Two nematode species from freshwater and marine fishes in Thailand, including Ascarophis scatophagi sp. nov. (Cystidicolidae) from Scatophagus argus (Scatophagidae). Acta Parasitologica 63: 89–98. [IF2017 = 1.039] Netherlands E.C., Cook C.A., Du Preez L.H., Vanhove M.P.M., Brendonck L., Smit N.J. (2017) Monophyly of the species of Hepatozoon (Adeleorina: Hepatozoidae) parasitising (African) anurans, with the description of three new species from hyperoliid frogs in South Africa. Parasitology 145: 1039–1050. [IF2017 = 2.511] Oros M., Uhrovič D., Scholz T. (2018) A new classification of Glaridacris Cooper, 1920 (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in north America, including erection of Pseudoglaridacris n.gen. The Journal of Parasitology 104: 60–69. [IF2017 = 1.395] Pantoja C., Scholz T., Luque J.L., Jones A. (2018) New genera and species of paramphistomes (Digenea: Paramphistomoidea: Cladorchiidae) parasitic in fishes from the Amazon basin in Peru. Systematic Parasitology 95: 611–624. [IF2017 = 1.273] Pakosta T., Vetešník L., Šimková A. (2018) A Long Temporal Study of Parasitism in Asexual-Sexual Populations of Carassius gibelio: Does the Parasite Infection Support Coevolutionary Red Queen Dynamics? BioMed Research International Volume 2018: Article ID 6983740. [IF2017 = 2.398] Patra S., Bartošová-Sojková P., Pecková H., Fiala I., Eszterbauer E., Holzer A.S. (2018) Biodiversity and host-parasite cophylogeny of Sphaerospora (sensu stricto) (Cnidaria: Myxozoa). Parasites & Vectors 11: 347. [IF2017 = 3.086] Přikrylová I., Mašová Š., Gelnar M., Matla M.M., Tavakol S., Luus-Powell W.J. (2018) Redescription of the genus Afrodiplozoon Khotenovski, 1981 and its only known species Afrodiplozoon polycotyleus (Paperna, 1973) (Monogenea: Diplozoidae) using a combined multidisciplinary approach. Parasitology International 67(2): 245–252. [IF2017 = 1.886] Rahmouni C., Vanhove M.P.M., Šimková A. (2018) Seven new species of Cichlidogyrus Paperna, 1960 (Monogenea: Dactylogyridae) parasitizing the gills of Congolese cichlids from northern Lake Tanganyika. PeerJ 6: e5604. [IF2017 = 2.158]

Reichard M., Lanés L.E.K., Polačik M., Blažek R., Vrtílek M., Godoy R. S., Maltchik L. (2018) Avian predation mediates size-specific survival in a Neotropical annual fish: a field experiment. Biological Journal of the Linnean Society 124: 56–66. [IF2017 = 2.358] Roche K., Kuta J., Sedláček I., Červenka R., Tomanová K., Jurajda P. (2018) First data on uranium uptake in three nototheniid fishes from Antarctica (James Ross Island), Chemosphere 211: 510–514. [IF2017 = 4.589] Scholz T., Barčák D., Oros M. (2018) The occurrence of the non-native tapeworm Khawia japonensis (Yamaguti, 1934) (Cestoda) in cultured common carp in the Czech Republic confirms its recent expansion in Europe. BioInvasions Records 7(3): 303–308. [IF2017 = 1.189] Scholz T., Tavakol S., Uhrová L., Brabec J., Přikrylová I., Mašová Š., Šimková A., Halajian A., Luus-Powell W.J. (2018) An annotated list and molecular data on larvae of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) from freshwater fishes in Africa. Systematic Parasitology 95: 567–590. [IF2017 = 1.273] Scholz T., Šimková A., Razanabolana J.R., Kuchta R. (2018) The first record of the invasive Asian fish tapeworm (Schyzocotyle acheilognathi) from an endemic cichlid fish in Madagascar. Helminthologia 55(1): 84–87. [IF2017 = 0.417] Simdyanov T.G., Paskerova G.G., Valigurová A., Diakin A., Kováčiková M., Schrével, J., Gillou L., Dobrovolskij A.A., Aleoshin V.V. (2018) First ultrastructural and molecular phylogenetic evidence from the blastogregarines, an early branching lineage of plesiomorphic Apicomplexa. Protist 169(5): 697–726. [IF2017 = 2.511] Smith C., Spence R., Reichard M. (2018) Sperm is a sexual ornament in rose bitterling. Journal of Evolutionary Biology doi: 10.1111/jeb.13357. [IF2017 = 2.483] Šimková A., Řehulková E., Rasoloariniaina J.R., Jorissen M.W.P., Scholz T., Faltýnková A., Mašová Š., Vanhove M.P.M. (2018). Transmission of parasites from introduced tilapias: a new threat to endemic Malagasy ichthyofauna. Biological Invasions 2018: 1–17. [IF2017 = 3.128] Tyml T., Dyková I. (2018) Sappinia sp. (Amoebozoa: Thecamoebida) and Rosculus sp. (SAR: Cercozoa) isolated from king penguin guano collected in the Subantarctic (South Georgia, Salisbury Plain) and their coexistence in culture. Journal of Eukaryotic Microbiology 65: 544–555. [IF2017 = 2.552] Tyml T., Lisnerová M., Kostka M., Dyková I. (2018) Current view on phylogeny within the genus Flabellula Schaeffer, 1926 (Amoebozoa: Leptomyxida). European Journal of Protistology 64: 40–53. [IF2017 = 2.426] Vanhove M.P.M., Briscoe A.G., Jorissen M.W.P., Littlewood D.T.J., Huyse T. (2018) The first next-generation sequencing approach to the mitochondrial phylogeny of African monogenean parasites (Platyhelminthes: Gyrodactylidae and Dactylogyridae). BMC Genomics 19: 520. [IF2017 = 3.753]

SUBMITTED in 2018 English C.J., Tyml T., Botwright N.A., Barnes A.C., Wynne J.W., Lima P.C., Cook M.T.A diversity of amoebae colonise the gills of farmed Atlantic salmon, Salmo salar with amoebic gill disease (AGD). European Journal of Protistology (submitted). Hulak B., Kvach Y. The finding of Pseudobacciger harengulae (Yamaguti, 1938) (Digenea: Faustulidae) in the Mediterranean horse mackerel (Trachurus mediterraneus (Steindachner, 1868)) (Actinopterygii: Carangidae) from the Gulf of Odessa, Black Sea, Ukraine. Vestnik Zoologii (in press). Jedličková L., Dvořák J., Hrachovinová I., Kašný M., Ulrychová L., Mikeš L. A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro. International Journal for Parasitology. (accepted). Jorissen M.W.P., Pariselle A., Huyse T., Vreven E.J., Snoeks J., Decru E., Kusters T., Wamuini Lunkayilakio S., Muterezi Bukinga F., Artois T., Vanhove M.P.M. Six new dactylogyrid species (Platyhelminthes: Monogenea) from the gills of cichlids (Teleostei: Cichliformes) from the Lower Congo Basin. Parasite. (accepted). Kvach Y., Matvienko N., Bryjová A., Ondračková M. Aquaculture as a possible vector in the spread of Posthodiplostomum centrarchi (Hoffman, 1958) (Digenea: Diplostomidae) in Europe. BioInvasion Records (in press). Kvach Y., Ondračková M., Drobiniak O., Zamorov V. The parasitisation of the monkey goby, Neogobius fluviatilis, in the localities with different range of salinity. Oceanological and Hydrobiological Studies (in press). Kvach Y., Ondračková M., Janáč M., Krasnovyd V., Seifertová M., Jurajda P. Parasites of round goby, Neogobius melanostomus, currently invading the River Elbe. Oceanological and Hydrobiological Studies (in press). Nezhybová V., Blažek R., Kašný M., Slamková D., Leontovyč R., Ondračková M. Morphological and molecular characterization of Apatemon sp. infecting killifish in Mozambique. Parasitology International (submitted). Ondračková M., Kvach Y., Martens A., Jurajda P. Limited parasite acquisition by non-native Lepomis gibbosus (Antinopterygii: Centrarchidae) at two ponds in the Upper Rhine basin, Germany. Journal of Helminthology (in press). Ondračková M., Fojtů J., Seifertová M., Kvach Y., Jurajda P. Non-native parasitic copepod Neoergasilus japonicus (Harada, 1930) utilizes non-native fish host Lepomis gibbosus (L.) in the floodplain of the River Dyje (Danube basin). Parasitology Research (accepted). Paskerova G.G., Miroliubova T.S., Diakin A., Kováčiková M., Valigurová A., Gillou L., Aleoshin V.V., Simdyanov T.G. (2018) Fine structure and molecular phylogeny of two marine gregarines, Selenidium pygospionis n. sp. and S. pherusae n. sp., with notes on the phylogeny of Archigregarinida (Apicomplexa). Protist (In Press). Pravdová M., Ondračková M., Přikrylová I., Blažek R., Mahmoud Z., Gelnar M. Dactylogyrids (Platyhelminthes: Monogenea) from Sudanese Labeo spp., with description of Dogielius sennarensis n. sp. and the redescription of Dogielius flosculus Guégan, Lambert & Euzet, 1989. Helminthologia (in press). Roudnický P., Vorel J., Ilgová J., Benovics M., Norek A., Mikeš L., Jedličková L., Potěšil D., Zdráhal Z., Dvořák J. Dalton J. Gelnar. M., Kašný M. Identification and partial characterization of a novel serpin from Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea). Parasite (submitted). Roumbedakis K., Krausova M., Tyml T., Di Cristo C. A perspective around cephalopods and their parasites, and suggestions on how to increase knowledge in the field. Frontiers in Physiology (submitted). Van Steenberge M.W., Raeymaekers J.A.M., Hablützel P.I., Vanhove M.P.M., Koblmüller S., Snoeks J. Delineating species along shifting shorelines: Tropheus (Teleostei, Cichlidae) from the southern subbasin of Lake Tanganyika. Frontiers in Zoology. (in press). Všetičková L., Mikl L., Adámek Z., Prášek V., Roche K., Jurajda P. The diet of reservoir perch before, during and after establishment of non-native tubenose goby. Knowledge and Management of Aquatic Ecosystems (in press).

2017 Acosta A.A., Franceschini L., Zago A.C., Scholz T., da Silva R.J. (2017) Six new species of Heteropriapulus (Monogenea: Dactylogyridae) from South American fishes with an amended diagnosis to the genus. Zootaxa 4290: 459– 482. [IF2017 = 0.931] Acosta A.A., González-Solís D., da Silva R.J. (2017) Spinitectus aguapeiensis n. sp. (Nematoda: Cystidicolidae) from Pimelodella avanhandavae Eigenmann (Siluriformes: Heptapteridae) in the River Aguapeı, Upper Parana River Basin, Brazil. Systematic Parasitology 94: 649–656. [IF2017 = 1.273] Alves P.V., de Chambrier A., Luque J.L., Scholz T. (2017) Reappraisal of Goezeella Fuhrmann, 1916 (Cestoda: Proteocephalidae), parasites of Neotropical catfishes (Siluriformes), with description of a new species from Pimelodella cristata (Heptapteridae). Revue Suisse de Zoologie 124: 335–350. [IF2017 = 0.759] Alves P.V., de Chambrier A., Luque J.L., Scholz T. (2017) Untangling convoluted taxonomy of Chambriella Rego, Chubb & Pavanelli, 1999 (Cestoda: Proteocephalidae), with erection of Riggenbachiella n. g. and the description of a new species from pimelodid catfishes in the Neotropical Region. Systematic Parasitology 94: 367–389. [IF2017 = 1.273] Alves P.V., de Chambrier A., Scholz T., Luque J.L. (2017) Annotated checklist of fish cestodes from South America. ZooKeys 650: 1–205. [IF2017 = 1.079] Arredondo N.J., Alves P.V., Gil de Pertierra A.A. (2017) A new genus of proteocephalid tapeworm (Cestoda) from the marbled swamp eel Synbranchus marmoratus Bloch (Synbranchiformes: Synbranchidae) in the River Paraná basin, Argentina. Folia Parasitologica 64: 015. [IF2017 = 1.505] Barčák D., Oros M., Hanzelová V., Scholz T. (2017) A synoptic review of Caryophyllaeus Gmelin, 1790 (Cestoda: Caryophyllidea), parasites of cyprinid fishes. Folia Parasitologica 64: 027. [IF2017 = 1.505] Benhamou F., Marzoug D., Boutiba Z., Kostadinova A., Pérez-del-Olmo A. (2017) Parasite communities in two sparid fishes from the Western Mediterranean: a comparative analysis based on samples from three localities of the Algerian coast. Helminthologia 54: 26–35. [IF2017 = 0.417] Born-Torrijos A, Holzer A.S., Raga J.A., Van Beest G.S., Yoneva A. (2017) Description of embryonic development and ultrastructure in miracidia of Cardiocephaloides longicollis (Digenea, Strigeidae) in relation to active host finding strategy in a marine environment. Journal of Morphology 278: 1137–1148. [IF2017 = 1.655] de Chambrier A., Pinacho-Pinacho M., Hernández-Orts J., Scholz T. (2017) A new genus and two new species of proteocephalidean tapeworms (Cestoda) from cichlid fish (Perciformes: Cichlidae) in the Neotropics. Journal of Parasitology 103: 83–89. [IF2017 = 1.395] Diakin A., Wakeman K., Valigurová A. (2017) Description of Ganymedes yurii sp. n. (Ganymedidae), a new gregarine species from the Antarctic amphipod Gondogeneia sp. (Crustacea). Journal of Eukaryotic Microbiology 64: 56–66. [IF2017 = 2.552] Fannes W., Vanhove M.P.M., Huyse T. (2017) Redescription of Cichlidogyrus tiberianus Paperna, 1960 and C. dossoui Douëllou, 1993 (Monogenea: Ancyrocephalidae), with special reference to the male copulatory organ. Systematic Parasitology 94(1): 133–144. [IF2017 = 1.273] Francová K., Seifertová M., Blažek R., Gelnar M., Mahmoud Z.N., Řehulková E. (2017) Quadriacanthus species (Monogenea: Dactylogyridae) from catfishes (Teleostei: Siluriformes) in eastern Africa: new species, new records and first insights into interspecific genetic relationships. Parasites & Vectors 10: 361. [IF2017 = 3.086] Georgieva S., Blasco-Costa I., Kostadinova A. (2017) Molecular characterisation of four echinostomes (Digenea: Echinostomatidae) from birds in New Zealand, with descriptions of Echinostoma novaezealandense n. sp. and Echinoparyphium poulini n. sp. Systematic Parasitology 94: 477–497. [IF2017 = 1.273] González-Solís D., Mariaux J. (2017) Orientatractis brycini sp. nov. (Nematoda: Atractidae) from characiform freshwater fishes in Gabon, Africa. Revue Suisse de Zoologie 124: 1–8. [IF2017 = 0.759] Hablützel P.I., Vanhove M.P.M., Deschepper P., Grégoir A.F., Roose A.K., Volckaert F.A.M., Raeymaekers J.A.M. (2017) Parasite escape through trophic specialization in a species flock. Journal of Evolutionary Biology 30 (7): 1437–1445. [IF2016 = 2.792] Hernández-Orts J.S., Hernández-Mena D.I., Alama-Bermejo G., Kuchta R., Jacobson K.C. (2017) Morphological and molecular characterisation of Aporocotyle margolisi Smith, 1967 (Digenea: Aporocotylidae) from the North Pacific hake Merluccius productus (Ayres) (Gadiformes: Merlucciidae) off Oregon, USA. Systematic Parasitology 94: 819–829. [IF2017 = 1.273] Ilgová J., Jedličková L., Dvořáková H., Benovics M., Mikeš L., Janda L., Vorel J., Roudnický P., Potěšil D., Zdráhal Z., Gelnar M., Kašný M. (2017) A novel type I cystatin of parasite origin with atypical legumain-binding domain, Scientific Reports 7: 17526. [IF2017 = 4.248] Janáč M., Bryja J., Ondračková M., Mendel J., Jurajda P. (2017) Genetic structure of three invasive gobiid species along the Danube-Rhine invasion corridor: similar distributions, different histories. Aquatic Invasions 12: 551–564. [IF2017 = 2.000] Jirsová D., Štefka J., Jirku M. (2017) Discordant population histories of host and its parasite: a role for ecological permeability of extreme environment? PLoS ONE 12: e0175286. [IF2017 = 2.859]

Kalogianni E., Kmentová N., Harris E., Zimmerman B., Giakoumi S., Chatzinikolaou Y., Vanhove M.P.M. (2017) Occurrence and effect of trematode parasite metacercariae on two endangered killifishes from Greece. Parasitology Research 116(11): 3007–3018. [IF2017 = 2.329] Koblmüller S., Nevado B., Makasa L., Van Steenberge M., Vanhove M.P.M., Verheyen E., Sturmbauer C., Sefc K.M. (2017) Phylogeny and phylogeography of Altolamprologus: ancient introgression and recent divergence in a rock-dwelling Lake Tanganyika cichlid genus. In: Koblmüller S., Albertson, R. C., Genner, M. J., Sefc, K. M., Takahashi, T. (eds.) Advances in Cichlid Research II: Behavior, Ecology and Evolutionary Biology. Hydrobiologia 791(1): 35–50. [IF2017 = 2.022] Konstanzová V., Koubková B., Kašný M., Ilgová J., Dzika E., Gelnar M. (2017) An ultrastructural study of the surface and attachment structures of Paradiplozoon homoion (Bychowsky & Nagibina, 1959) (Monogenea: Diplozoidae). Parasites & Vectors 10: 1–10. [IF2017 = 3.086] Kostka M., Lares-Jiménez L.F., Tyml T., Dyková I. (2017) Copromyxa laresi n. sp. (Amoebozoa: Tubulinea) and transfer of Cashia limacoides (Page, 1967) to Copromyxa Zopf, 1885. Journal of Eukaryotic Microbiology 64: 78–87. [IF2017 = 2.552] Kováčiková M., Simdyanov T.G., Diakin A., Valigurová A. (2017) Structures related to attachment and motility in the marine eugregarine Cephaloidophora cf. communis (Apicomplexa). European Journal of Protistology 59: 1– 13. [IF2017 = 2.426] Kudlai O., Kostadinova A., Pulis E.E., Tkach V.V. (2017) The Psilostomidae Looss, 1900 (sensu stricto) (Digenea: Echinostomatoidea): description of three new genera and a key to the genera of the family. Systematic Parasitology 94: 21–33. [IF2017 = 1.273] Kudlai O., Oros M., Kostadinova A., Georgieva S. (2017) Exploring the diversity of Diplostomum (Digenea: Diplostomidae) in fishes from the River Danube using mitochondrial DNA barcodes. Parasites & Vectors 10: 592. [IF2017 = 3.086] Kvach Y., Bryjová A., Sasal P., Winkler H.M. (2017) A revision of the genus Aphalloides (Digenea: Cryptogonimidae), parasites of European brackish water fishes. Parasitology Research 116(7): 1973–1980. [IF2017 = 2.329] Kvach Y., Janáč M., Nehring S., Ondračková M., Jurajda P. (2017) Parasite communities and infection levels of the invasive Chinese sleeper Perccottus glenii (Actinopterygii: Odontobutidae) from the Naab River basin, Germany. Journal of Helminthology 91(6): 703–710. [IF2017 = 1.262] Kvach Y., Jurajda P., Bryjová A., Trichkova T., Ribeiro F., Přikrylová I., Ondračková M. (2017) European distribution for metacercariae of the North American digenean Posthodiplostomum cf. minimum centrarchi (Strigeiformes: Diplostomidae). Parasitology International 66(5): 635–642. [IF2017 = 1.886] Kvach Y., Ondračková M., Bryjová A., Jurajda P. (2017) Parasites as biological tags of divergence in Central European gudgeon populations (Actinopterygii: Cyprinidae: Gobioninae). 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[IF2017 = 1.394] Monnens M., Artois T., Vanhove M.P.M. (2017) Syndesmis aethopharynx (Umagillidae, Rhabdocoela, Platyhelminthes) from the sea urchin Paracentrotus lividus: first record from the Eastern Mediterranean, phylogenetic position and intraspecific morphological variation. Parasitology International 66(6): 848–858. [IF2017 = 1.886] Moravec F., Bakenhaster M., Leone E.H. (2017) Redescription of Philometra margolisi Moravec, Vidal-Martínez et Aguirre-Macedo, 1995 (Nematoda: Philometridae), a gonad-infecting parasite of the red grouper Epinephelus morio (Serranidae) in the Gulf of Mexico. Acta Parasitologica 62: 412–421. [IF2017 = 1.039] Moravec F., Beveridge I. (2017) Lobocapillaria austropacifica n. g., n. sp. (Nematoda: Capillariidae) from the obtuse barracuda Sphyraena obtusata Cuvier (Sphyraenidae, Perciformes) off eastern Australia. Systematic Parasitology 94: 547–556. [IF2017 = 1.273] Moravec F., Chaabane A., Neifar L., Gey D., Justine J.-L. (2017) Species of Philometra (Nematoda, Philometridae) from fishes off the Mediterranean coast of Africa, with a description of Philometra rara n. sp. from Hyporthodus haifensis and a molecular analysis of Philometra saltatrix from Pomatomus saltatrix. Parasite 24: 8. [IF2017 = 2.069] Moravec F., Ghanmi N., Chaabane A., Gargouri L., Justine J.-L. (2017) Redescription of Philometra filiformis (Stossich, 1896) (Nematoda: Philometridae), a gonad-infecting parasite of the common pandora Pagellus erythrinus

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(2017) Description and experimental transmission of Tetracapsuloides vermiformis n. sp. (Cnidaria: Myxozoa) and guidelines for describing malacosporean species including reinstatement of Buddenbrockia bryozoides n. comb. (syn. Tetracapsula bryozoides). Parasitology 144: 497–511. [IF2017 = 2.713] Polačik M., Janáč M. (2017) Costly defense in a fluctuating environment-sensitivity of annual Nothobranchius fishes to predator kairomones. Ecology and Evolution 7(12): 4289–4298. [IF2017 = 2.389] Přikrylová I., Shinn A.P., Paladini G. (2017) Description of Citharodactylus gagei n. gen. et n. sp. (Monogenea: Gyrodactylidae) from the moon fish, Citharinus citharus (Geoffroy Saint-Hilaire), from Lake Turkana, Kenya. Parasitology Research 116(1): 281–292. [IF2017 = 2.329] Přikrylová I., Smit N.J., Gelnar M. (2017) Description of Afrogyrodactylus ardae sp. n. (Monogenea: Gyrodactylidae) from Rhabdalestes septentrionalis (Characifromes: Alestidae) in the Niokolo-Koba National Park, Senegal. 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(2016) An optimised multi-host trematode life cycle: fishery discards enhance trophic parasite transmission to scavenging birds. International Journal for Parasitology 46: 745–753. [IF2016 = 4.242] Born-Torrijos A., Raga J.A., Holzer A.S. (2016) Trematode maturation patterns in a migratory snail host: what happens during upshore residency in a Mediterranean lagoon? Parasitology Research 115: 575–585. [IF2016 = 2.098] Chambouvet A., Valigurová A., Mesquita L., Richards T.A., Jirků M. (2016) Nematopsis temporariae (Gregarinasina, Apicomplexa, Alveolata) intracellular infectious agent of tadpole livers. Environmental Microbiology Reports 8(5): 675–679. [IF2016 = 3.038] de Chambrier A., Scholz T. (2016) An emendation of the generic diagnosis of the monotypic Glanitaenia (Cestoda: Proteocephalidae), with notes on the geographical distribution of G. osculata, a parasite of invasive wels catfish. Revue Suisse de Zoologie 123: 1–9. 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[IF2016 = 1.181] Forte-Gil D., Holzer A.S., Pecková H., Bartošová-Sojková P., Peñalver J., Dolores E.M., Muñoz P. (2016) Molecular and morphological identification of Cardicola (Trematoda: Aporocotylidae) eggs in hatchery-reared and migratory Atlantic bluefin tuna (Thunnus thynnus L.). Aquaculture 450: 58–66. [IF2016 = 1.893] Hablützel P.I., Grégoir A.F., Vanhove M.P.M., Volckaert F.A.M., Raeymaekers J.A.M. (2016) Weak link between dispersal and parasite community differentiation or immunogenetic divergence in two sympatric cichlid fishes. Molecular Ecology 25: 5451–5466. [IF2016 = 5.317] Hartigan A., Estensoro I., Vancová M., Bílý T., Patra S., Eszterbauer E., Holzer A.S. (2016) New cell motility model observed in parasitic cnidarian Sphaerospora molnari (Myxozoa: Myxosporea) blood stages in fish. Scientific Reports 6: 39093. [IF2016 = 5.228] Hartigan A., Wilkinson M., Gower D.J., Streicher J.W., Holzer A.S., Okamura B. (2016) Myxozoan infections of caecilians demonstrate broad host specificity and indicate a link with human activity. International Journal for Parasitology 46: 375–381. [IF2016 = 4.242] Jedličková L., Dvořáková H., Kašný M., Ilgová J., Potěšil D., Zdráhal Z., Mikeš L. (2016) Major acid endopeptidases of the blood-feeding monogenean Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae). Parasitology 143: 494–506. [IF2016 = 2.560] Jurajda P., Adámek Z., Roche K., Mrkvová M., Štarhová D., Prášek V., Zukal J. (2016) Carp feeding activity and habitat utilisation in relation to supplementary feeding in a semi-intensive aquaculture pond, Aquaculture International 24: 1627–1640. [IF2016 = 1.279] Jurajda P., Adámek Z., Janáč M., Roche K., Mikl L., Rederer L., Zapletal T., Koza V., Špaček J. (2016) Use of multiple fish-removal methods during biomanipulation of a drinking water reservoir – Evaluation of the first four years. Fisheries Research 173: 101–108. [IF2016 = 2.223] Jurajda P., Roche K., Sedláček I., Všetičková L. (2016) Assemblage characteristics and diet of fish in the shallow coastal waters of James Ross Island, Antarctica. Polar Biology 39: 2299–2309. Jurajda P., Pavlov I. (2016) Rediscovery of Rutilus virgo in the River Dyje, Czech Republic. Folia Zoologica 65: 98–100. [IF2016 = 0.986] Kmentová N., Gelnar M., Koblmüller S., Vanhove M.P.M. (2016) Deep-water parasite diversity in Lake Tanganyika: description of two new monogenean species from benthopelagic cichlid fishes. Parasites & Vectors 9: 426. [IF2016 = 3.234]

Kmentová N., Gelnar M., Koblmüller S., Vanhove M.P.M. (2016) First insights into the diversity of gill monogeneans of 'Gnathochromis' and Limnochromis (Teleostei, Cichlidae) in Burundi: do the parasites mirror host ecology and phylogenetic history? PeerJ 4: e1629. [IF2016 = 2.225] Kmentová N., Gelnar M., Mendlová M., Van Steenberge M., Koblmuller S., Vanhove M. P. M. (2016) Reduced host- specificity in a parasite infecting non-littoral Lake Tanganyika cichlids evidenced by intraspecific morphological and genetic diversity. Scientific Reports 6: 39605. [IF2016 = 4.259] Konstanzová V., Koubková B., Kašný M., Ilgová J., Dzika E., Gelnar M. (2016) Excretory system of representatives from family Diplozooidae (Monogenea). Parasitology Research 115: 1493–1500. [IF2016 = 2.098] Kudlai O., Cribb T.H., Cutmore S.C. (2016) A new species of microphallid (Trematoda: Digenea) infecting a novel host family, the Muraenidae, on the northern Great Barrier Reef, Australia. 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[IF2016 = 1.038] Mendoza-Franco E.F., Mendoza-Palmero C.A., Scholz T. (2016) New species of Ameloblastella Kritsky, Mendoza- Franco & Scholz, 2000 and Cosmetocleithrum Kritsky, Thatcher & Boeger, 1986 (Monogenea: Dactylogyridae) infecting the gills of catfishes (Siluriformes) from the Peruvian Amazonia. Systematic Parasitology 93: 847–862. [IF2016 = 1.181] Moravec F., Adlard R. (2016) Redescription of Rhabdochona papuanensis (Nematoda: Thelazioidea), a parasite of rainbow fishes (Melanotaenia spp.); the first record of the speciesof Rhabdochona in Australia. Acta Parasitologica 61: 820–827. [IF2016 = 1.160] Moravec F., Ali A.H., Abed J.M., Shaker S.J. (2016) New records of philometrids (Nematoda: Philometridae) from marine fishes off Iraq, with the erection of two new species and the first description of the male of Philometroides eleutheronemae Moravec & Manoharan, 2013. Systematic Parasitology 93: 129–144. [IF2016 = 1.181] Moravec F., Bakenhaster M.D., Adams D.H. (2016) Two new species of Philometra Costa, 1845 (Nematoda: Philometridae) from Mycteroperca spp. (Serranidae) in the North-West Atlantic and northern Gulf of Mexico, USA. Systematic Parasitology 93: 479–491. [IF2016 = 1.181] Moravec F., Barton D.P. (2016) New tissue-dwelling species of Philometra Costa, 1845 and Philometroides Yamaguti, 1935 (Nematoda: Philometridae) from marine perciform fishes off the northern coast of Australia. Systematic Parasitology 93: 623–637. [IF2016 = 1.181] Moravec F., Gey D., Justine J.-L. (2016) Nematode parasites of four species of Carangoides (Osteichthyes: Carangidae) in New Caledonian waters, with a description of Philometra dispar n. sp. (Philometridae). Parasite 23: 40. [IF2016 = 2.545] Moravec F., Chaabane A., Justine J.-L., Neifar L. (2016) Two gonad-infecting species of Philometra (Nematoda: Philometridae) from groupers (Serranidae) off Tunisia, with a key to Philometra species infecting serranid gonads. Parasite 23: 8. [IF2016 = 2.545] Moravec F., Chaabane A., Neifar L., Gey D., Justine J.-L. (2016) Descriptions of Philometra aenei n. sp. and P. tunisiensis n. sp. (Nematoda: Philometridae) from Epinephelus spp. off Tunisia confirm a high degree of host specificity of gonad-infecting species of Philometra Costa, 1845 in groupers (Serranidae). Systematic Parasitology 93: 115–128. [IF2016 = 1.181]

Moravec F., Manoharan J. (2016) Philometra dissimilis n. sp. from the ovary of Johnius belangerii (Sciaenidae) and other new records of philometrids (Nematoda: Philometridae) from fishes of the Bay of Bengal, India. Helminthologia 53: 133–141. [IF2016 = 0.563] Moravec F., Pachanawan A., Kamchoo K. (2016) Redescription of two species of cystidicolid nematodes (Spirurina: Cystidicolidae) from Notopterus notopterus (Osteichthyes) in Thailand. Acta Parasitologica 61: 278–290. [IF2016 = 1.160] Moravec F., van Rensburg C.J., Van As L. (2016) Larvae of Contracaecum sp. (Nematoda: Anisakidae) in the threatened freshwater fish Sandelia capensis (Anabantidae) in South Africa. Diseases of Aquatic Organisms 120: 251–254. [IF2016 = 1.549] Moravec F., Scholz T. (2016) Helminth parasites of the lesser great cormorant Phalacrocorax carbo sinensis from two nesting regions in the Czech Republic. Folia Parasitologica 63: 022. [IF2016= 1.082] Moreira J., Scholz T., Luque J.L. (2016) A new species of Diaphorocleidus (Monogenea: Ancyrocephalinae) from the gills of Argonectes robertsi (Characiformes) and new records of dactylogyrids parasitic on fishes from the Xingu River, Amazon Basin, Brazil. Zoologia 33: e20160022. [IF2016 = 0.642] Oros M., Brabec J., Kuchta R., Choudhury A., Scholz T. (2016) A synoptic review of Promonobothrium Mackiewicz, 1968 (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in North America, with description of two new species. Folia Parasitologica 63: 008. [IF2016= 1.082] Papežíková I., Mareš J., Vojtek L., Hyršl P., Marková Z., Šimková A., Bartoňková J., Navrátil S., Palíková M. (2016) Seasonal changes in immune parameters of rainbow trout (Oncorhynchus mykiss), brook trout (Salvelinus fontinalis) and brook trout x Arctic charr hybrids (Salvelinus fontinalis x Salvelinus alpinusxalpinus). Fish & Shellfish Immunology 57: 400–405. [IF2016 = 3.407] Paschoal F., Scholz T., Tavares-Dias M., Luque L. (2016) Dactylogyrids (Monogenea) parasitic on cichlids from northern Brazil, with description of two new species of Sciadicleithrum and new host and geographical records. Acta Parasitologica 61: 158–164. [IF2016 = 1.160] Paskerova G.G., Frolova E.V., Kováčiková M., Panfilkina T.S., Mesentsev E.S., Smirnov A.V., Nassonova E.S. (2016) Metchnikovella dogieli sp. n. (Microsporidia: Metchnikovellida), a parasite of archigregarines Selenidium sp. from polychaetes Pygospio elegans. Protistology 10(4): 148–157. Pérez-del-Olmo A., Kostadinova A., Gibson D. I. (2016). The Mediterranean: high discovery rates for a well-studied trematode fauna. Systematic Parasitology 93: 249–256. [IF2016 = 1.181] Pizzetti I., Schulz F., Tyml T., Fuchs B.M., Amann R., Horn M., Fazi S. (2016) Chlamydial seasonal dynamics and isolation of the “Candidatus Neptunochlamydia vexilliferae” from a Tyrrhenian coastal lake. Environmental Microbiology 18: 2405–2417. [IF2016 = 5.109] Pornruseetairatn S., Kino H., Shimazu T., Nawa Y., Scholz T., Ruangsittichai J., Tanomsing Saralamba N., Thaenkham U. (2016) A molecular phylogeny of Asian species of the genus Metagonimus (Digenea) – small intestinal flukes – based on representative Japanese populations. Parasitology Research 115: 1123–1130. [IF2016 = 2.329] Raphahlelo M.E., Přikrylová I., Matla M.M., Theron J., Luus-Powell W.J. (2016) A revised description of Synodontella zambezensis Douëllou et Chishawa, 1995 (Monogenea: Ancyrocephalidae) from the gills of Synodontis zambezensis (Siluriformes: Mochokidae) from South Africa. Helminthologia 53(4): 363–371. [IF2016 = 0.563] Řežucha R., Reichard M. (2016) The association between personality traits, morphological traits and alternative mating behaviour in male Endler’s guppies, Poecilia wingei. Ethology 122: 456–467. [IF2016 = 1.359] Seifertová M., Jarkovský J., Šimková A. (2016) Does the parasite-mediated selection drive the MHC class IIB diversity in wild populations of European chub (Squalius cephalus)? Parasitology Research 115(4): 1401–1415. [IF2016 = 2.329] Scholz T., Besprozvannykh V.V., Boutorina T.E., Choudhury A., Cribb T.H., Ermolenko A.V., Faltýnková A., Shedko M.B., Shimazu T., Smit N.J. (2016) Trematode diversity in freshwater fishes of the Globe I: ‘Old World’. Systematic Parasitology 93: 257–269. [IF2016 = 1.181] Schrével J., Valigurová A., Prensier G., Chambouvet A., Florent I., Guillou L. (2016) Ultrastructure of Selenidium pendula, the type species of archigregarines, and phylogenetic relations to other marine Apicomplexa. Protist 167(4): 339–368. [IF2016 = 2.673] Šlapanský L., Jurajda P., Janáč M. (2016) Early life stages of exotic gobiids as new hosts for unionid glochidia. Freshwater Biology 61: 979–990. [IF2016 = 3.174] Tihlaříková E., Neděla V., Mašová Š. (2016) Environmental Scanning Electron Microscopy as a Useful Tool for Taxonomical Documentation of Parasitical Helminths. Microscopy And Microanalysis 22(S3): 1178–1179. [IF2015 = 1.730] Tkach V.V., Kudlai O., Kostadinova A. (2016) Molecular phylogeny and systematics of the Echinostomatoidea Looss, 1899 (Platyhelminthes: Digenea). International Journal for Parasitology 46: 171–185. [IF2016 = 3.370] Truter M., Přikrylová I., Malherbe W. Smit N.J. (2016) First report of metazoan parasites from the cichlid, Pseudocrenilabrus philander and cyprinid, Enteromius paludinosus in a Ramsar wetland, South Africa. African Journal of Aquatic Sciences 41(4): 499–503. [IF2016 = 0.806]

Tyml T., Skulinová K., Kavan J., Ditrich O., Kostka M., Dyková, I. (2016) Heterolobosean amoebae from Arctic and Antarctic extremes: 18 novel strains of Allovahlkampfia, Vahlkampfia and Naegleria. European Journal of Protistology 56: 119–133. [IF2016 = 2.419] Tyml T., Kostka M., Ditrich O., Dyková, I. (2016) Vermistella arctica n. sp. nominates the genus Vermistella as a candidate for taxon with bipolar distribution. Journal of Eukaryotic Microbiology 63: 210–219. [IF2016 = 2.490] Vanhove M.P.M., Hablützel P.I., Pariselle A., Šimková A., Huyse T., Raeymaekers J.A.M. (2016) Cichlids: a host of opportunities for evolutionary parasitology. Trends in Parasitology 32(10): 820–832. [IF2016 = 5.292] Vanhove M.P.M., Kovačić M., Zogaris S. (2016) A distinct island population of threatened freshwater fish: to split or lump? Hydrobiologia 777(1): 79–93. [IF2016 = 2.127] Vrtílek M., Reichard M. (2016) Patterns of morphological variation among populations of the widespread annual killifish Nothobranchius orthonotus are independent of genetic divergence and biogeography. Journal of Zoological Systematics and Evolutionary Research 54: 289–298. [IF2016 = 2.667] Vrtílek M., Reichard M. (2016) Female fecundity traits in wild populations of African annual fish: the role of the aridity gradient. Ecology and Evolution 6: 5921–5931. [IF2016 = 2.626] Wünnemann H., Holzer A.S., Pecková H., Bartošová-Sojková P., Eskens U., Lierz M. (2016) Repatriation of an old fish host as an opportunity for myxozoan parasite diversity: the example of the allis shad, Alosa alosa (Clupeidae), in the Rhine. Parasites & Vectors 9: 505. [IF2016 = 3.234] Zahradníčková P., Barson M., Luus-Powell W.J., Přikrylová I. (2016) Species of Gyrodactylus von Nordmann, 1832 (Platyhelminthes: Monogenea) from cichlids in Zambezi and Limpopo river basins from Zimbabwe and South Africa: evidence for unexplored species richness. Systematic Parasitology 93(7): 679–700. [IF2016 = 1.181] Zapletal T., Adámek Z., Jurajda P., Roche K., Všetičková L., Mareš J. (2016) Consumption of plant material by perch (Perca fluviatilis). Folia Zoologica 65: 95–97. [IF2016 = 0.986]

2015 Alves P.V., de Chambrier A., Scholz T., Luque J.L. (2015) A new genus and species of proteocephalidean tapeworm (Cestoda), first cestode found in the driftwood catfish Tocantinsia piresi (Siluriformes: Auchenipteridae) in South America. Folia Parasitologica 62: 006. [IF2015 = 1.271] Antar R., Georgieva S., Gargouri L., Kostadinova A. (2015) Molecular evidence for the existence of species complexes within Macvicaria Gibson & Bray, 1982 (Digenea: Opecoelidae) in the western Mediterranean, with descriptions of two new species. Systematic Parasitology 91: 211–229. [IF2015 = 1.316] Ash A., de Chambrier A., Shimazu T., Ermolenko A., Scholz T. (2015) An annotated list of the species of Gangesia Woodland, 1924 (Cestoda: Proteocephalidea), parasites of catfishes in Asia, with new synonyms and a key to their identification. Systematic Parasitology 91: 13–33. [IF2015 = 1.316] Bartošová-Sojková P., Kodádková A., Pecková H., Kuchta R., Reed C.C. (2015) Morphology and phylogeny of two new species of Sphaeromyxa Thélohan, 1892 (Cnidaria: Myxozoa) from marine fish (Clinidae and Trachichthyidae). Parasitology 142: 660–674. [IF2015 = 2.560] Bartošová-Sojková P., Oppenheim R., Soldati-Favre D., Lukeš J. (2015) Epicellular apicomplexans: parasites “on- the-way-in”. PLoS Pathogens 11: e1005080. [IF2015 = 7.562] Brabec J., Waeschenbach A., Scholz T., Littlewood D.T.J., Kuchta R. (2015) Molecular phylogeny of the Bothriocephalidea (Cestoda): molecular data challenge morphology-based classification. International Journal for Parasitology 45: 761–771. [IF2015 = 3.234] de Chambrier A., Kuchta R., Scholz T. (2015) Tapeworms (Cestoda: Proteocephalidea) of teleost fishes from the Amazon River in Peru: additional records as an evidence of unexplored species diversity. Revue Suisse de Zoologie 122: 149–163. [IF2015 = 0.431] de Chambrier A., Waeschenbach A., Fisseha M., Scholz T., Mariaux J. (2015) A large 28S rDNA-based phylogeny confirms the limitations of established morphological characters for classification of proteocephalidean tapeworms (Platyhelminthes, Cestoda). ZooKeys 500: 25–59. [IF2015 = 0.938] Dallarés S., Pérez-del-Olmo A., Carrassón M., Kuchta R. (2015) Morphological and molecular characterisation of Ditrachybothridium macrocephalum Rees, 1959 (Cestoda: Diphyllidea) from Galeus melastomus Rafinesque in the Western Mediterranean. Systematic Parasitology 92: 45–55. [IF2015 = 1.316] Dogga S.K., Bartošová-Sojková P., Lukeš J., Soldati-Favre D. (2015) Phylogeny, morphology, and metabolic and invasive capabilities of epicellular fish coccidium Goussia janae. Protist 166: 659–676. [IF2015 = 3.045] Faltýnková A., Georgieva S., Soldánová M., Kostadinova A. (2015) A re-assessment of species diversity within the ‘revolutum’ group of Echinostoma Rudolphi, 1809 (Digenea: Echinostomatidae) in Europe. Systematic Parasitology 90: 1–25. [IF2015 = 1.316] Fannes W., Vanhove M.P.M., Huyse T., Paladini G. (2015) A scanning electron microscope technique for studying the sclerites of Cichlidogyrus. Parasitology Research 114(5): 2031–2034. [IF2015 = 2.098] Fiala I., Hlavničková M., Kodádková A., Freeman M.A., Bartošová-Sojková P., Atkinson S.D. (2015) Evolutionary origin of Ceratonova shasta and phylogeny of the marine myxosporean lineage. Molecular Phylogenetics and Evolution 86: 75–89. [IF2015 = 4.122] Forte-Gil D., Holzer A.S., Pecková H., Bartošová-Sojková P., Peñalver J., Dolores E.M., Muñoz P. (2015) Molecular and morphological identification of Cardicola (Trematoda: Aporocotylidae) eggs in hatchery-reared and migratory Atlantic bluefin tuna (Thunnus thynnus L.). Aquaculture 450: 58–66. [IF2015 = 1.893] Gil de Pertierra A.A., Arredondo N.J., Kuchta R., Incorvaia I.S. (2015) A new species of Bothriocephalus Rudolphi, 1808 (Eucestoda: Bothriocephalidea) from the channel bull blenny Cottoperca gobio (Günther) (Perciformes: Bovichtidae) on the Patagonian shelf off Argentina. Systematic Parasitology 90: 247–256. [IF2015 = 1.316] Grégoir A.F., Hablützel P.I., Vanhove M.P.M., Pariselle A., Bamps J., Volckaert F.A.M., Raeymaekers J.A.M. (2015) A link between host dispersal and parasite diversity in two sympatric cichlid fishes of Lake Tanganyika. Freshwater Biology 60: 323–335. [IF2015 = 2.738] Hanzelová V., Oros M., Barčák D., Miklisová D., Kirin D., Scholz T. (2015) Morphological polymorphism in tapeworms: redescription of Caryophyllaeus laticeps (Cestoda: Caryophyllidea) and characterization of its morphotypes from different fish hosts. Systematic Parasitology 90: 177–190. [IF2015 = 1.316] Havlátová L., Ondračková M., Přikrylová I. (2015) Monogenean parasites of pumpkinseed sunfish Lepomis gibbosus in Durance River, France. Helminthologia 52: 323–330. [IF2015 = 0.695] Isbert W., Montero F.E., Carrassón M., Gonzáles-Solís D. (2015) Dichelyne (Cucullanellus) romani n.sp. (Nematoda: Cucullanidae) in notacanthid fishes from the Northeast Atlantic and Western Mediterranean. Systematic Parasitology 91: 35-47. [IF2015 = 1.316] Huyse T., Vanhove M.P.M., Mombaerts M., Volckaert F.A.M., Verreycken H. (2015) Parasite introduction with an invasive goby in Belgium: double trouble? Parasitology Research 114(7): 2789–2793. [IF2015 =2.098] Jurajda P., Adámek Z., Valová Z., Janáč M., Roche K. (2015) Fish community and fisheries management at Brno reservoir following revitalisation measures. Folia Zoologica. 64(2): 112–122. [IF2015 = 0.662]

Kašparová E., Van de Putte A.P., Marshall C., Janko K. (2015) Lifestyle and Ice: The relationship between ecological specialization and response to Pleistocene climate change. PLoS ONE 10(11). DOI: 10.1371/journal.pone.0138766. [IF2015 = 3.451] Keppeler F.W., Lanés L.E.K., Rolon A.S., Stenert C., Lehmann P., Reichard M., Maltchik L. (2015) The morphology- diet relationship and its role in the coexistence of two species of annual fishes. Ecology of Freshwater Fish 24: 77–90. [IF2015 = 2.039] Kičinjaová M. L., Blažek R., Gelnar M., Řehulková E. (2015) Annulotrema (Monogenea: Dactylogyridae) from the gills of African tetras (Characiformes: Alestidae) in Lake Turkana, Kenya, with descriptions of four new species and a redescription of A. elongata Paperna and Thurston, 1969. Parasitology Research 114: 4107– 4120. [IF2015 = 2.098] Kodádková A., Bartošová-Sojková P., Holzer A.S., Fiala I. (2015) Bipteria vetusta n. sp. – an old parasite in an old host: tracing the origin of myxosporean parasitism in vertebrates. International Journal for Parasitology 45: 269–276. [IF2015 = 3.872] Konečná, M., Janáč, M., Roche, K., Jurajda, P. (2015) Variation in life-history traits between a newly established and long-established population of non-native pumpkinseed, Lepomis gibbosus (Actinopterygii: Perciformes: Centrarchidae). Acta Ichthyologica et Piscatoria 45: 385–392. [IF2015 = 0.756] Konstanzová V., Koubková B., Kašný M., Ilgová J., Dzika E., Gelnar M. (2015) Ultrastructure of the digestive tract of Paradiplozoon homoion (Monogenea). Parasitology Research 114: 1485–1494. [IF2015 = 2.330] Kudlai O., Cutmore S.C., Cribb T.H. (2015) Morphological and molecular data for three species of the Microphallidae (Trematoda: Digenea) in Australia, including the first descriptions of the cercariae of Maritrema brevisacciferum and Microphallus minutus. Folia Parasitologica 62: 053. [IF2015 = 1.271] Kudlai O., Kostadinova A., Pulis E.E., Tkach V.V. (2015) A new species of Drepanocephalus Dietz, 1909 (Digenea: Echinostomatidae) from the double-crested cormorant Phalacrocorax auritus (Lesson) (Aves: Phalacrocoracidae) in North America. Systematic Parasitology 90: 221–230. [IF2015 = 1.316] Lisitsyna O., Scholz T., Kuchta R. (2015) Sharpilosentis peruviensis n. g., n. sp. (Acanthocephala: Palaeacanthocephala) from freshwater catfishes (Siluriformes) in the Amazonia. Systematic Parasitology 91: 147–155. [IF2015 = 1.316] Marchiori N.C., Pariselle A., Pereira Jr. J., Agnèse J.-F., Durand J.-D., Vanhove M.P.M. (2015) A comparative study of Ligophorus uruguayense and Ligophorus saladensis (Monogenea, Ancyrocephalidae) from Mugil liza (Teleostei, Mugilidae) in southern Brazil. Folia Parasitologica 62: 024. [IF2015 = 1.147] Mendoza-Palmero C.A., Blasco-Costa I., Scholz T. (2015) Molecular phylogeny of Neotropical monogeneans (Platyhelminthes: Monogenea) from catfishes (Siluriformes). Parasites & Vectors 8: 164. [IF2015 = 3.430] Messu Mandeng F.D., Bilong Bilong C.F., Pariselle A., Vanhove M.P.M., Bitja Nyom A.R., Agnèse J.-F. (2015) A phylogeny of Cichlidogyrus species (Monogenea, Dactylogyridea) clarifies a host switch between fish families and reveals an adaptive component to attachment organ morphology of this parasite genus. Parasites & Vectors 8: 582. [IF2015 = 3.430] Mikhailov K.V., Tikhonenkov D.V., Janouškovec J., Diakin A., Ofitserov M.V., Mylnikov A.P., Aleshin V.V. (2015) Primary structure of 28S rRNA gene confirms monophyly of free-living heterotrophic and phototrophic apicomplexans (Alveolata). Biochemistry 80(11): 1492–1499. [IF2015 = 3.174] Moravec F., Barton D.P. (2015) Two gonad-infecting species of Philometra (Nematoda: Philometridae) from marine fishes off the northern Australia. Parasite 22: 4. [IF2015 = 1.781] Moravec F., Diggles B.K. (2015) A new gonad-infecting species of Philometra, P. barnesi sp. n. (Nematoda: Philometridae), from the marine fish Pomadasys argenteus (Haemulidae) off the northern coast of Australia. Parasitology Research 114: 4121–4126. [IF2015 = 2.027] Moravec F., Diggles B.K. (2015) Philometra mirabilis sp. n. (Nematoda: Philometridae), a new gonad-infecting parasite from the freshwater fish Cichla mirianae (Cichlidae) in Brazilian Amazon. Parasitology Research 114: 1929–1932. [IF2015 = 2.027] Moravec F., Halajian A., Tavakol S., Nyagura I., Luus-Powell W.J. (2015) Philometroides khalili n. sp., a new philometrid nematode (Philometridae) from the operculum of the cyprinid fish Labeo rosae in Zimbabwe. Helminthologia 52: 113–117. [IF2015 = 0.602] Moravec F., Jirků M. (2015) Two Procamallanus (Spirocamallanus) species (Nematoda: Camallanidae) from freshwater fishes in the Lower Congo River. Acta Parasitologica 60: 226-–233. [IF2015 = 1.293] Moravec F., Justine J.-L. (2015) Anisakid nematodes (Nematoda: Anisakidae) from the marine fishes Plectropomus laevis Lacépède (Serranidae) and Sphyraena qenie Klunzinger (Sphyraenidae) off New Caledonia, including two new species of Hysterothylacium Ward & Magath, 1917. Systematic Parasitology 92: 181–195. [IF2015 = 1.316] Moravec F., Justine J.-L. (2015) New records of species of Philometra (Nematoda: Philometridae) from marine fishes off New Caledonia, including P. cephalopholidis sp. n. from Cephalopholis sonnerati (Serranidae). Parasitology Research 114: 3223–3228. [IF2015 = 2.027]

Moravec F., Scholz T. (2015) Macroparasites and their communities of the European eel Anguilla anguilla (Linnaeus) in the Czech Republic. Folia Parasitologica 62: 033. [IF2015 = 1.271] Moravec F., Tedesco P. (2015) Redescription of Philometra globiceps (Rudolphi, 1819) (Nematoda: Philometridae), the type species of Philometra Costa, 1845, including the first data obtained by SEM. Parasitology Research 114: 4413-4420. [IF2015 = 2.027] Moravec F., Van As L.L. (2015) Procamallanus (Procamallanus) spp. (Nematoda: Camallanidae) in fishes of the Okavango River, Botswana, including the description of P. (P.) pseudolaeviconchus n. sp. parasitic in Clarias spp. (Clariidae) from Botswana and Egypt. Systematic Parasitology 90: 137–149. [IF2015 = 1.316] Moravec F., Van As L.L. (2015) Procamallanus (Spirocamallanus) spp. (Nematoda: Camallanidae) from fishes of the Okavango River, Botswana, including P. (S.) serranochromis n. sp. parasitic in Serranochromis spp. (Serranidae). Systematic Parasitology 90: 151–164. [IF2015 = 1.316] Moravec F., Van As L.L. (2015) Studies on some spirurids (Nematoda: Spirurida) from fishes of the Okavango River, Botswana. Systematic Parasitology 91: 119–138. [IF2015 = 1.316] Moravec F., Van As L.L. (2015) Studies on ascaridid, oxyurid and enoplid nematodes (Nematoda) from fishes of the Okavango River, Botswana. Folia Parasitologica 62: 039. [IF2015 = 1.271] Moravec F., Kamchoo K., Pachanawan A. (2015) New nematode species, Orientatractis mekongensis n. sp. (Atractidae) and Neosynodontisia suratthaniensis n. g., n. sp. (Pharyngodonidae) from freshwater fishes in Thailand. Systematic Parasitology 92: 197–209. [IF2015 = 1.316] Moreira J., Scholz T., Luque J.L. (2015) First data on the parasites of Hoplias aimara (Characiformes): description of two new species of gill monogeneans (Dactylogyridae). Acta Parasitologica 60: 254–560. [IF2015 = 1.293] Nezhybová V., Mašová Š. (2015) Basic epidemiological data on metazoan parasites of notothenioid fish off James Ross Island (Prince Gustav Channel, Weddell Sea), Antarctica. Czech Polar Reports 5: 44–54. [IF2015 = 0.950] O’Dwyer K., Faltýnková A., Georgieva S., Kostadinova A. (2015) An integrative taxonomic investigation of the diversity of digenean parasites infecting the intertidal snail Austrolittorina unifasciata Gray, 1826 (Gastropoda: Littorinidae) in Australia. Parasitology Research 114: 2381–2397. [IF2015 = 2.027] Ondračková M., Hudcová I., Dávidová M., Adámek Z., Kašný M., Jurajda P. (2015). Non-native gobies facilitate the transmission of Bucephalus polymorphus (Trematoda). Parasites & Vectors 19: 382. [IF2015 = 3.430] Pariselle A., Muterezi Bukinga F., Van Steenberge M., Vanhove M.P.M. (2015) Ancyrocephalidae (Monogenea) of Lake Tanganyika: IV: Cichlidogyrus parasitizing species of Bathybatini (Teleostei, Cichlidae): reduced host- specificity in the deepwater realm? In: Koblmüller S., Albertson R.C., Genner M.J., Sefc K.M. & Takahashi T. (eds.) Advances in cichlid research: Behavior, ecology and evolutionary biology. Hydrobiologia 748(1): 99– 119. [IF2015 = 2.275] Pariselle A., Van Steenberge M., Snoeks J., Volckaert F.A.M., Huyse T., Vanhove M.P.M. (2015) Ancyrocephalidae (Monogenea) of Lake Tanganyika: does the Cichlidogyrus parasite fauna of Interochromis loocki (Teleostei, Cichlidae) reflect its host’s phylogenetic affinities? Contributions to Zoology 84(1): 25–38. [IF2015 = 1.656] Pereira F.B., Tavares L.E.R., Paiva F., Scholz T., Luque J. (2015) A morphological and molecular study of Spectatus spectatus (Kathlaniidae), including redescription of the species and amendment of genus diagnosis. Journal of Parasitology 101: 468–475. [IF2015 = 1.394] Pereira F.B., Tavares L.E.R., Scholz T., Luque J.L. (2015) A morphological and molecular study of two species of Raphidascaroides Yamaguti, 1941 (Nematoda: Anisakidae), parasites of doradid catfish (Siluriformes) in South America, including description of R. moraveci n. sp. Systematic Parasitology 91: 49–61. [IF2015 = 1.316] Pérez-i-García D., Constenla M., Carrasón M., Montero F.E., Soler-Membrives A., González-Solís D. (2015) Raphidascaris (Raphidascaris) macrouri n.sp. (Nematoda: Anisakidae) from two deep-sea macrourid fishes in the Western Mediterranean: Morphological and molecular characterisations. Parasitology International 64: 345-354. [IF2015 = 1.860] Poddubnaya L.G., Kuchta R., Bristow G.A., Scholz T. (2015) Ultrastructure of the anterior organ and posterior funnel-shaped canal of Gyrocotyle urna Wagener, 1852 (Cestoda: Gyrocotylidea). Folia Parasitologica 62: 027. [IF2015 = 1.271] Řežucha R., Reichard M. (2015) Strategic exploitation of fluctuating asymmetry in male Endler’s guppy courtship displays is modulated by social environment. Journal of Evolutionary Biology 28: 356–367. [IF2015 = 2.990] Scholz T., Oros M., Choudhury A., Brabec J., Waeschenbach A. (2015) New circumscrition of freshwater fish parasites Monobothrium Diesing, 1863 and Promonobothrium Mackiewicz, 1968 (Cestoda: Caryophyllidea) using morphological and molecular evidence. Journal of Parasitology 101: 29–36. [IF2015 = 1.394] Scholz T., Tavakol S., Halajian A., Luus-Powell W.J. (2015) The invasive fish tapeworm Atractolytocestus huronensis (Cestoda), a parasite of carp, colonises Africa. Parasitology Research 114: 3521–3524. [IF2015 = 2.027] Schulz F., Tyml T., Pizzetti I., Dyková I., Fazi S., Kostka M., Horn, M. (2015) Marine amoebae with cytoplasmic and perinuclear symbionts deeply branching in the Gammaproteobacteria. Scientific Reports 5: 13381. [IF2015 = 5.622]

Simdyanov T.G., Diakin A., Vladimir V., Aleoshin V.V. (2015) Ultrastructure and 28S rDNA phylogeny of two gregarines: Cephaloidophora cf. communis and Heliospora cf. longissima with remarks on gregarine morphology and phylogenetic analysis. Acta Protozoologica 54: 241–262. [IF2015 = 1.821] Smith C., Phillips A., Reichard M. (2015) Cognitive ability is heritable and predicts the success of an alternative mating tactic. Proceedings of the Royal Society B: Biological Sciences 282: 20151046. [IF2015 = 5.167] Valigurová A., Paskerova G.G., Diakin A., Kováčiková M., Simdyanov T.G. (2015) Protococcidian Eleutheroschizon duboscqi, an unusual apicomplexan interconnecting gregarines and cryptosporidia. PLoS ONE 10(4): e0125063. [IF2015 = 3.451] Van Steenberge M., Pariselle A., Huyse T., Volckaert F.A.M., Snoeks J., Vanhove M.P.M. (2015) Morphology, molecules, and monogenean parasites: an example of an integrative approach to cichlid biodiversity. PLoS ONE 10(4): e0124474. [IF2015 = 3.234] Vanhove M.P.M., Pariselle A., Van Steenberge M., Raeymaekers J.A.M., Hablützel P.I., Gillardin C., Hellemans B., Breman F.C., Koblmüller S., Sturmbauer C., Snoeks J., Volckaert F.A.M., Huyse T. (2015) Hidden biodiversity in an ancient lake: phylogenetic congruence between Lake Tanganyika tropheine cichlids and their monogenean flatworm parasites. Scientific Reports 5: 13669 [IF2015 = 5.578]

2014 Barčák D., Oros M., Hanzelová V., Scholz T. (2014) Phenotypic plasticity in Caryophyllaeus brachycollis Janiszewska, 1953 (Cestoda: Caryophyllidea): does fish host play a role? Systematic Parasitology 88: 153–166. [IF2014 = 1.336] Bartošová-Sojková P., Hrabcová M., Pecková H., Patra S., Kodádková A., Jurajda P., Tyml T., Holzer A.S. (2014) Hidden diversity and evolutionary trends in malacosporean parasites (Cnidaria: Myxozoa) identified using molecular phylogenetics. International Journal for Parasitology 44: 565–577. [IF2014 = 3.404] Bazsaloviczová E., Králová-Hromadová I., Brabec J., Hanzelová V., Oros M., Scholz T. (2014) Conflict between morphology and molecular data: a case of the genus Caryophyllaeus (Cestoda, Caryophyllidea), monozoic tapeworms of cyprinid fishes. Folia Parasitologica 61: 347–354. [IF2014 = 1.147] Blasco-Costa I., Faltýnková A., Georgieva S., Skirnisson K., Scholz T., Kostadinova A. (2014) Fish pathogens near the Arctic Circle: molecular, morphological and ecological evidence for unexpected diversity of Diplostomum (Digenea: Diplostomidae) in Iceland. International Journal for Parasitology 44: 703–715. [IF2014 = 3.872] Born-Torrijos A., Poulin R., Raga J.A., Holzer A.S. (2014) Estimating trematode prevalence in snail hosts using a single-step duplex PCR: how badly does cercarial shedding underestimate infection rates? Parasites & Vectors 7: 243. [IF2014 = 3.251] Chang C.H., Li F., Shao K. T., Lin Y.S., Morosawa T., Kim S., Koo H., Kim W., Lee J. S., He S.P., Smith C., Reichard M., Miya M., Sado T., Uehara K., Lavoué S., Chen W.J., Mayden R.L. (2014) Phylogenetic relationships of Acheilognathidae (Cypriniformes: Cyprinoidea) as revealed from evidence of both nuclear and mitochondrial gene sequence variation: Evidence for necessary taxonomic revision in the family and the identification of cryptic species. Molecular Phylogenetics and Evolution 81: 182-194. [IF2014 = 4.319] de Chambrier A., Scholz T., Kuchta R. (2014) Taxonomic status of Woodland’s enigmatic tapeworms (Cestoda: Proteocephalidea) from Amazonian catfishes: back to museum collections. Systematic Parasitology 87: 1– 19. [IF2014 = 1.336] Faltýnková A., Georgieva S., Kostadinova A., Blasco-Costa I., Scholz T., Skírnisson, K. (2014) Diplostomum von Nordmann, 1832 (Digenea: Diplostomidae) in the sub-Arctic: descriptions of the larval stages of six species discovered recently in Iceland. Systematic Parasitology 89: 195–213. [IF2014 = 1.336] Francová K., Ondračková M. (2014) Effect of habitat conditions on parasite infection in 0+ juvenile perch (Perca fluviatilis L.) in two Czech reservoirs. Hydrobiologia 721: 57–66. [IF2014 = 2.212] Georgieva S., Faltýnková A., Brown R., Blasco-Costa I., Soldánová M., Sitko J., Scholz T., Kostadinova A. (2014) Echinostoma ʻrevolutumʼ (Digenea: Echinostomatidae) species complex revisited: species delimitation based on novel molecular and morphological data gathered in Europe. Parasites & Vectors 7: 520. [IF2014 = 3.430] González-Solís D., Carrassón M., Pérez-del-Olmo A. (2014) Capillostrongyloides morae sp. n. (Nematoda: Capillariidae) from deep-sea fish (Teleostei, Moridae) in the western Mediterranean Sea. Folia Parasitologica 61: 63–68. [IF2014 = 1.147] González-Solís D., Chavan S.P., Kannewad P., Gyananath G. (2014) A new species of Rhabdochona Railliet, 1916 (Nematoda: Rhabdochonidae) from cyprinid fishes in the Western Ghats Region, India. Systematic Parasitology 87: 273–281. [IF2014 = 1.336] Hablützel P.I., Vanhove M.P.M., Grégoir A.F., Hellemans B., Volckaert F.A.M, Raeymaekers J.A.M. (2014) Intermediate number of MHC class IIB length variants relates to enlarged perivisceral fat deposits in the blunt-head cichlid Tropheus moorii. Journal of Evolutionary Biology 27(10): 2177–2190. [IF2014 = 3.532] Holzer A.S., Hartigan A., Patra S., Pecková H., Eszterbauer E. (2014) Molecular fingerprinting of the myxozoan community in common carp suffering Swim Bladder Inflammation (SBI) identifies multiple etiological agents. Parasites & Vectors 7: 398. [IF2014 = 3.251] Keppeler F.W., Lanés L.E.K., Rolon A.S., Stenert C., Lehmann P., Reichard M., Maltchik L. (2014) The morphology-diet relationship and its role in the coexistence of two species of annual fishes. Ecology of Freshwater Fish doi: 10.1111/eff.12127. [IF2014 = 1.590] Kodádková A., Dyková I., Tyml T., Ditrich O., Fiala I. (2014) Myxozoa in high Arctic: survey on the central part of Svalbard archipelago. International Journal for Parasitology: Parasites and Wildlife 3: 41–56. [IF2014 = 2.777] Kuchta R., Pearson R., Scholz T., Ditrich O., Olson P.D. (2014) Spathebothriidea: survey of species, scolex and egg morphology, and interrelationships of a non-segmented, relictual tapeworm group (Platyhelminthes: Cestoda). Folia Parasitologica 61: 331–346. [IF2014 = 1.147] Liao C.P., Yu D., Chen Y.Y., Reichard M., Liu H.Z. (2014) Reproductive behaviour of female rosy bitterling Rhodeus ocellatus in response to a female-biased operational sex ratio. Behaviour 151: 755-768. [IF2014 = 1.401] Marzoug D., Rima M., Boutiba Z., Georgieva S., Kostadinova A., Pérez-del-Olmo, A. (2014) A new species of Saturnius Manter, 1969 (Digenea: Hemiuridae) from Mediterranean mullet (Teleostei: Mugilidae). Systematic Parasitology 87: 127–134. [IF2014 = 1.336]

Mašová Š., Baruš V., Seifertová M., Malala J, Jirků M. (2014) Redescription and molecular characterisation of Dujardinascaris madagascariensis and a note on D. dujardini (Nematoda: Heterocheilidae), parasites of Crocodylus niloticus, with a key to Dujardinascaris spp. in crocodilians. Zootaxa 3893(3): 261–276. [IF2014 = 0.996] Mendlová M., Šimková A. (2014) Evolution of host specificity in monogeneans parasitizing African cichlid fish. Parasites & Vectors 7: 69 [IF2014 = 3.251] Michálková V., Ondračková M. (2014) Experimental evidence for parasite-induced overwinter mortality in juvenile Rhodeus amarus. Journal of Fish Biology 84: 1377–1388. [IF2014 = 1.734] Mikhailov K.V., Janouškovec J., Tikhonenkov D.V., Mirzaeva G.S., Diakin A., Simdyanov T.G., Mylnikov A.P., Keeling P.J., Aleoshin V.V. (2014) A complex distribution of elongation family GTPases EF1A and EFL in basal alveolate lineages. Genome Biology and Evolution 6(9):2361–2367. [IF2014 = 4.486] Moravec F., Ali A.H. (2014) Additional observations on Philometra spp. (Nematoda: Philometridae) in marine fishes off Iraq, with the description of two new species. Systematic Parasitology 87: 259–271. [IF2014 = 1.336] Moravec F., Bakenhaster M., Fajer-Avila E.J. (2014) Three new gonad-infecting species of Philometra (Nematoda: Philometridae) parasitic in Lutjanus spp. (Lutjanidae) in the northern Gulf of Mexico off Florida, USA. Folia Parasitologica 61: 355–369. [IF2014 = 1.147] Moravec F., Diggles B.K. (2014) Philometrid nematodes (Philometridae) from marine fishes off the northern coast of Australia, including three new species. Folia Parasitologica 61: 37–54. [IF2014 = 1.147] Moravec F., Diggles B.K. (2014) Two new gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) from marine fishes off the northern coast of Australia. Systematic Parasitology 89: 33–44. [IF2014 = 1.336] Moravec F., Diggles B.K., Barnes L., Macbeth W. (2014) Buckleyella ornata n.sp. (Nematoda: Philometridae) from the abdominal cavity of the talang queenfish Scomberoides commersonnianus (Perciformes: Carangidae) off the northern coast of Australia. Helminthologia 51: 230-235. [IF2014 = 0.678] Moravec F., Justine J.-L. (2014) Capillaria plectropomi n.sp. (Nematoda: Cillariidae) a new intestinal parasite of the leopard coral grouper Plectropomus leopardus (Serranidae) off New Caledonia. Parasite 21: 76. [IF2014 = 1.092] Moravec F., Justine J.-L. (2014) Philometrids (Nematoda: Philometridae) in carangid and serranid fishes off New Caledonia, including three new species. Parasite 21: 21. [IF2014 = 1.092] Moravec F., Jirků M. (2014) Rhabdochona spp. (Nematoda: Rhabdochonidae) from fishes in the Central African Republic, including three new species. Folia Parasitologica 61: 157–172. [IF2014 = 1.147] Moravec F., Jirků M. (2014) Dujardinascaris mormyropsis n. sp. (Nematoda: Anisakidae) from the osteoglossiform fish Mormyrops anguilloides (Linnaeus) (Mormyridae) Central Africa. Systematic Parasitology 88: 55–62. [IF2014 = 1.336] Moravec F., Khosheghbal M., Pazooki J. (2014) Dichelyne (Dichelyne) spinigerus sp. nov. (Nematoda: Cucullanidae) from the marine fish Otolithes ruber (Sciaenidae) off Iran and first description of the male of Philometra otolithi Moravec et Manoharan, 2013 (Nematoda: Philometridae). Acta Parasitologica 59: 229–237. [IF2014 = 0.905] Moravec F., Manoharan J. (2014) Two new gonad-infecting species of Philometra (Nematoda: Philometridae) parasitic in Lutjanus spp. (Osteichthyes: Lutjanidae) in the Bay of Bengal, India. Parasitology Research 113: 3299–3307. [IF2014 = 2.098] Moravec F., Manoharan J. (2014) Gonad-infecting species of Philometra (Nematoda: Philometridae) from groupers Epinephelus spp. (Osteichthyes: Serranidae) in the Bay of Bengal, India. Acta Parasitologica 59: 596–605. [IF2014 = 0.905] O’Dwyer K., Blasco-Costa I., Poulin R., Faltýnková A. (2014) Four marine digenean parasites of Austrolittorina spp. (Gastropoda: Littorinidae) in New Zealand: morphological and molecular data. Systematic Parasitology 89: 133–152. [IF2014 = 1.336] Pérez-del-Olmo A., Dallarés S., Carrassón M., Kostadinova A. (2014) A new species of Bathycreadium Kabata, 1961 (Digenea: Opecoelidae) from Phycis blennoides (Brünnich) (Gadiformes: Phycidae) in the western Mediterranean. Systematic Parasitology 88: 233–244. [IF2014 = 1.336] Pérez-del-Olmo A., Georgieva S., Pula H., Kostadinova A. (2014) Molecular and morphological evidence for three species of Diplostomum (Digenea: Diplostomidae), parasites of fishes and fish-eating birds in Spain. Parasites & Vectors 7: 502. [IF2014 = 3.430] Přikrylová I., Luus-Powell W. (2014) Revision of the genus Afrogyrodactylus Paperna, 1968 (Monogenea: Gyrodactylidae) with description of two new species, A. girgifae sp. n. and A. kingi sp. n., from geographically distant localities. Folia Parasitologica 61(6): 529–536. [IF2014 = 1.147] Reichenbacher B., Reichard M. (2014) Otoliths of five extant species of the annual killifish Nothobranchius from the East African savannah. PLoS ONE 9: e112459. [IF2014 = 3.530]

Řehulková E., Musilová N., Gelnar M. (2014) Annulotrema (Monogenea: Dactylogyridae) from Hydrocynus brevis (Günther) (Characiformes: Alestidae) in Segenal, with descriptions of two new species and remarks on Annulotrema pikei. Parasitology Research 113: 3273–3280. [IF2014 = 2.098] Scholz T., Choudhury A. (2014) Parasites of freshwater fishes in North America: why so neglected? Journal of Parasitology 100: 26–45. [IF2014 = 1.227] Scholz T., Oros M., Bazsalovicsová E., Brabec J., Waeschenbach A., Xi B.-W., Aydogdu A., Besprozvannykh V., Shimazu T., Králová-Hromadová I., Littlewood D.T.J. (2014) Molecular evidence of cryptic diversity in Paracaryophyllaeus (Cestoda: Caryophyllidea), parasites of loaches (Cobitidae) in Eurasia, including description of P. vladkae n. sp. Parasitology International 63: 841–850. [IF2014 = 1.859] Selbach C., Soldánová M., Georgieva S., Kostadinova A., Kalbe M., Sures, B. (2014) Morphological and molecular data for larval stages of four species of Petasiger Dietz, 1909 (Digenea: Echinostomatidae) with an updated key to the known cercariae from the Palaearctic. Systematic Parasitology 89: 153–166. [IF2014 = 1.336] Tolárová S., Dávidová M., Šimková A., Flajšhans M., Hyršl P. (2014) The seasonal changes of innate immunity of tench, Tinca tinca (L.) with different ploidy level. Aquaculture 432(1): 46–52. [IF2014 = 2.120] Valigurová A., Michalková V., Koník P., Dindo M.L., Gelnar M., Vaňhara J. (2014): Penetration and encapsulation of larval endoparasitoid, Exorista larvarum (Diptera: Tachinidae) in the factitious host Galleria mellonella (Lepidoptera: Pyralidae). Bulletin of Entomological Research 104(2): 203-212. [IF2014 = 2.058] Valová Z., Janáč M., Švanyga J., Jurajda P. (2014) Structure of 0+ juvenile fish assemblages in the modified upper stretch of the River Elbe, Czech Republic. Czech Journal of Animal Science 59: 35–44. [IF2014 = 0.871] Vanhove M.P.M., Economou A.N., Zogaris S., Giakoumi S., Zanella D., Volckeart F.A.M., Huyse T. (2014) The Gyrodactylus (Monogenea, Gyrodactylidae) parasite fauna of freshwater sand gobies (Teleostei, Gobioidei) in their centre of endemism, with description of seven new species. Parasitology Research 113(2): 653–668. [IF2014 = 2.098] Yoneva A., Kuchta R., Scholz T. (2014) First study of vitellogenesis of the broad fish tapeworm Diphyllobothrium latum (Cestoda, Diphyllobothriidea), a human parasite with extreme fecundity. Parasitology International 63: 747–753. [IF2014 = 1.859] Zapletal T., Mareš J., Jurajda P., Všetičková L. (2014) The food of roach, Rutilus rutilus (Actinopterygii: Cypriniformes: Cyprinidae), in a biomanipulated water supply reservoir. Acta Ichthyologica et Piscatoria 44: 15–22. [IF2014 = 0.691] Zikmundová J., Georgieva S., Faltýnková A., Soldánová M., Kostadinova A. (2014) Species diversity of Plagiorchis Lühe, 1899 (Digenea: Plagiorchiidae) in lymnaeid snails from freshwater ecosystems in central Europe revealed by molecules and morphology. Systematic Parasitology 88: 37–54. [IF2014 = 1.336]

2013 Adámek Z., Zahrádková S., Jurajda P., Bernardová I., Jurajdová Z., Janáč M., Němejcová D. (2013) The response of benthic macroinvertebrate and fish assemblages to human impact along the lower stretch of the rivers Morava and Dyje (Danube basin, Czech Republic). Croatian Journal of Fisheries 71: 93–115. Alama-Bermejo G., Šíma R., Raga J.A., Holzer A.S. (2013) Understanding myxozoan infection dynamics in the sea: seasonality and transmission of Ceratomyxa puntazzi. International Journal for Parasitology 43: 771–780. [IF2013 = 3.404] Bartáková V., Reichard M., Janko K., Polačik M., Blažek R., Reichwald K., Cellerino A., Bryja J. (2013) Strong population genetic structuring in an annual fish, Nothobranchius furzeri, suggests multiple savannah refugia in southern Mozambique. BMC Evolutionary Biology 13: 196. [IF2013 = 3.410] Bartošová P., Fiala I., Cinková M., Jirků M., Caffara M., Fioravanti M.L., Atkinson S.D., Bartholomew J.L., Holzer A.S. (2013) Sphaerospora sensu stricto: taxonomy, diversity and evolution of a unique lineage of myxosporeans (Myxozoa). Molecular Phylogenetics and Evolution 68: 93–105. [IF2013 = 4.287] Baruš V., Šimková A., Prokeš M., Peňáz M., Vetešník L. (2013) Heavy metals in two host-parasite systems: tapeworm vs. fish. Acta Veterinaria Brno 81, 313–317. [IF2013 = 0.537] Brandão M., Luque J.L., Scholz T., Kostadinova A. (2013) New records and descriptions of digeneans in the Magellanic penguin Spheniscus magellanicus (Aves: Sphenisciformes) on the coast of Brazil. Systematic Parasitology 85: 79–98. [IF2013 = 1.035] Chibwana F.D., Blasco-Costa I., Georgieva S., Hosea K. M., Nkwengulila G., Scholz T., Kostadinova A. (2013) A first insight into the barcodes for African diplostomids (Digenea: Diplostomidae): brain parasites in Clarias gariepinus (Siluriformes: Clariidae). Infection, Genetics and Evolution 17: 62–70. [IF2013 = 3.264] Civáňová K., Mustafa C., Koubková, B. (2012) The molecular and morphometrical decscription of a new diplozoid species from the gills of the Garra rufa (Heckel, 1843) (Cyprinidae) from Turkey – including a commentary on taxonomic division of Diplozoidae. Parasitology Research 112: 3053–3062. [IF2013 = 1.035] Dallarés S., Georgieva S., Kostadinova A., Carrassón M., Gibson D.I., Pérez-del-Olmo A. (2013) Morphometric and molecular characterisation of specimens of Lepidapedon Stafford, 1904 (Digenea: Lepidapedidae) from the deep-sea fish Mora moro (Risso) (Teleostei: Moridae) in the western Mediterranean. Systematic Parasitology 85: 243–253. [IF2013 = 1.035] Dyková I., Kodádková A., de Buron I., Fiala I., Roumillat W.A. (2013) Sinuolinea infections in the urinary system of Cynoscion species (Sciaenidae) and the search for the phylogenetic position of the type species of Sinuolinea Davis, 1917 (Myxozoa: Myxosporea). International Journal for Parasitology: Parasites and Wildlife 2: 10–17. [IF2013 = 2.777] Eszterbauer E., Sipos D., Forró B., Bartošová P., Holzer A.S. (2013) Molecular characterization of Sphaerospora molnari (Myxozoa), the agent of gill sphaerosporosis in common carp (Cyprinus carpio carpio). Diseases of Aquatic Organisms 104: 59–67. [IF2013 = 1.586] Francová K., Ondračková M. (2013) Overwinter body condition, mortality and parasite infection in two size classes of 0+ year juvenile European bitterling Rhodeus amarus. Journal of Fish Biology 82: 555–568. [IF2013 = 1.734] Georgieva S., Soldánová M., Pérez-del-Olmo A., Dangel D.R., Sitko J., Sures B., Kostadinova A. (2013) Molecular prospecting for European Diplostomum (Digenea: Diplostomidae) reveals cryptic diversity. International Journal for Parasitology 43: 57–72. [IF2013 = 3.404] Holzer A., Bartošová P, Pecková H., Tyml T., Atkinson S., Bartholomew J., Sipos D., Eszterbauer E., Dyková I. (2013) Who’s who in renal sphaerosporids (Bivalvulida: Myxozoa) from common carp, Prussian carp and goldfish – molecular identification of cryptic species, blood stages and new members of Sphaerospora sensu stricto. Parasitology 140: 46–60. [IF2013 = 2.350] Levron C., Yoneva A., Kalbe M. (2013) Spermatological characters in the diphyllobothriidean Schistocephalus solidus (Cestoda). Acta Zoologica (Stockholm) 94: 240–247. [IF2013 = 1.296] Mašová Š., Baruš V. (2013) Redescription of cystacanths of Corynosoma pseudohamanni Zdzitowiecki, 1984 (Acanthocephala: Polymorphidae) from paratenic fish hosts. Folia Parasitologica 60(2): 167–176. (IF2013 = 2.515) Molecular Ecology Resources Primer Development Consorcium, Cecilia Agosstini et al. (…Gettová L.,…. Šimková A.,….) (2013) Permanent Genetic Resources added to Molecular Ecology Reseources Database 1 April 2013- 31 May 2013. Molecular Ecology Resources 13: 966–968. [IF2013 = 7.432] Moravec F., Ali A.H. (2013) Philometra johnii sp. nov. (Nematoda, Philometridae), a new gonad-infecting philometrid from the sin croaker Johnius dussumieri (Cuvier) (Perciformes, Sciaenidae) from marine waters of Iraq. Acta Parasitologica 58: 263–268. [IF2013 = 0.965] Moravec F., Bakenhaster M. (2013) Two new gonad-infecting philometrids (Nematoda: Philometridae) from the yellowedge grouper Hyporthodus flavolimbatus (Serranidae) and the great northern tilefish Lopholatilus chamaeleonticeps (Malacanthidae) in the northern Gulf of Mexico. Systematic Parasitology 86: 113–123. [IF2013 = 1.035]

Moravec F., Bakenhaster M., de Buron I. (2013) A new gonad-infecting species of Philometra (Nematoda: Philometridae) from the Atlantic Spanish mackerel Scomberomorus maculatus (Scombridae) off the Atlantic coast of Florida and South Carolina. Journal of Parasitology 99: 290–296. [IF2013 = 1.258] Moravec F., de Buron I. (2013) A synthesis of our current knowledge of philometrid nematodes, a group of increasingly important fish parasites. Folia Parasitologica 60: 81–101. [IF2013 = 1.211] Moravec F., de Buron I., Measures L. (2013) First description of the gravid female of Philometra rubra (Leidy, 1856) (Nematoda: Philometridae), a parasite of the abdominal cavity of temperate basses Morone spp. (Moronidae: Perciformes) in North America. Journal of Parasitology 99: 496–500. [IF2013 = 1.258] Moravec F., Charo-Karisa H., Jirků M. (2013) The morphology and systematics of Rhabdochona paski Baylis, 1928 (Nematoda: Rhabdochonidae), a widespread parasite of freshwater fishes in Africa. Systematic Parasitology 85: 55–63. [IF2013 = 1.035] Moravec F., Khosheghbal M., Pazooki J. (2013) Two philometrids (Nematoda: Philometridae) infecting the tigertooth croaker Otolithes ruber (Bloch & Schneider) (Teleostei: Sciaenidae) off Iran, including erection of a new genus. Systematic Parasitology 86: 33–41. [IF2013 = 1.035] Moravec F., Kuchta R. (2013) Description of two new nematode species, parasites of the Mississippi paddlefish Polyodon spathula (Acipenseriformes: Polyodontidae). Journal of Parasitology 99: 680–685. [IF2013 = 1.258] Moravec F., Manoharan J. (2013) Gonad-infecting philometrids (Nematoda: Philometridae) including four new species from marine fishes off the eastern coast of India. Folia Parasitologica 60: 105–122. [IF2013 = 1.211] Moravec F., Pachanawan A., Kamchoo K. (2013) Rhabdochona (Rhabdochona) hypsibarbi n. sp. (Nematoda: Rhabdochonidae) from the freshwater cyprinid fish Hypsibarbus wetmorei (Smith) in northeast Thailand. Journal of Parasitology 99: 297–302. [IF2013 = 1.258] Moravec F., Sheeba S., Kumar A.B. (2013) Observations on nematodes from the Indonesian shortfin eel Anguilla bicolor bicolor McClelland in India, including a revalidation of Heliconema ahiri Karve, 1941 (Physalopteridae). Acta Parasitologica 58: 496–503. [IF2013 = 0.965] Moravec F., Taraschewski H., Weyl O.L.F. (2013) Redescription of Heliconema africanum (Linstow, 1899) n. comb. (Nematoda: Physalopteridae), a nematode parasite of freshwater eels (Anguilla spp.) in South Africa. Systematic Parasitology 85: 263–269. [IF2013 = 1.035] Repullés-Albelda A., Kostadinova A., Raga J.A., Montero F. (2013) Seasonal population dynamics of Zeuxapta seriolae (Monogenea: Heteraxinidae) parasitising Seriola dumerili (Carangidae) in the Western Mediterranean. Veterinary Parasitology 193: 163–171. [IF2013 = 2.545] Řehulková E., Mendlová M., Šimková A. (2013) Two new species of Cichlidogyrus (Monogenea: Dactylogyridae) parasitizing the gills of African cichlid fishes (Perciformes) from Senegal: morphometric and molecular characterization. Parasitology Research 112: 1399–1410. [IF2013 = 2.535] Šimková A., Civáňová K., Gettová L., Gilles A. (2013) Genomic porosity between invasive Chondrostoma nasus and endangered endemic Parachondrostoma toxostoma (Cyprinidae): the evolution of MHC IIB genes. PLoS ONE 8(6): e65883. [IF2013 = 4.005] Šimková A., Serbielle C., Pariselle A., Vanhove M.P.M., Morand S. (2013). Speciation in Thaparocleidus (Monogenea: Dactylogyridae) parasitizing Asian pangasiid catfishes. BioMed Research International Volume 2013: Article ID 353956. [IF2013 = 3.168] Valová Z., Hudcová H., Roche K., Svobodová J., Bernardová I., Jurajda P. (2013) No relationship found between mercury and lead concentrations in muscle and scales of chub Squalius cephalus L. Environmental Monitoring and Assessment 185: 3359–3368. [IF2013 = 1.679] Vanhove M.P.M, Tessens B., Schoelinck C., Jondelius U., Littlewood D.T., Artois T., Huyse T. (2013) Problematic barcoding in flatworms: A case-study on monogeneans and rhabdocoels (Platyhelminthes). ZooKeys 365: 355–379. [IF2013 = 1.029] Vetešník L., Halačka K., Šimková A. (2013) The effect of ploidy and temporal changes in the biochemical profile of gibel carp (Carassius gibelio): a cyprinid fish species with dual reproductive strategies. Fish Physiology and Biochemistry 39: 171–180. [IF2013 = 1.676] Xi, B.-W., Oros M., Wang G.-T., Scholz T., Xi J. 2013. Khawia abbottinae sp. n. (Cestoda: Caryophyllidea) from the Chinese false gudgeon Abbottina rivularis (Cyprinidae: Gobioninae) in China: morphological and molecular data. Folia Parasitologica 60: 141–148. [IF2013 = 1.211] Yera H., Kuchta R., Brabec J., Peyron F., Dupouy-Camet J. (2013) First identification of eggs of the Asian fish tapeworm Bothriocephalus acheilognathi (Cestoda: Bothriocephalidea) in human stool. Parasitology International 62: 268–271. [IF2013 = 2.111]

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2012 Ash A., Scholz T., de Chambrier A., Brabec J., Oros M., Kar P.K., Chavan S.P., Mariaux J. (2012) Revision of Gangesia (Cestoda: Proteocephalidea) in the Indomalayan region: morphology, molecules and surface ultrastructure. PLoS ONE 7: e46421. [IF2012 = 3.730] Blažek R., Ondračková M., Bímová Vošlajerová B., Vetešník L., Petrášová I., Reichard M. (2012) Fish diversity in the Niokolo Koba National Park, middle Gambia River basin, Senegal. Ichthyological Exploration of Freshwaters 23: 263–272. [IF2012 = 1.648] Born-Torrijos A., Kostadinova A., Raga J.A., Holzer A.S. (2012) Molecular and morphological identification of larval opecoelids (Digenea: Opecoelidae) parasitising prosobranch snails in a Western Mediterranean lagoon. Parasitology International 61: 450–460. [IF2012 = 2.302] Brabec J., Scholz T., Kraľová-Hromadová I., Bazsalovicsová E., Olson P.D. (2012) Substitution saturation and nuclear paralogs of commonly employed phylogenetic markers in the Caryophyllidea, an unusual group of non- segmented tapeworms (Platyhelminthes). International Journal for Parasitology 42: 259–267. [IF2012 = 3.637] Cárdenas M.Q., Moravec F., Fernandes B.M., Morais A.M. (2012) A new species of Philometra Costa, 1845 (Nematoda: Philometridae) from the freshwater fish (red piranha) Pygocentrus nattereri Kner (Characidae) in Amazonia, Brazil. Systematic Parasitology 83: 137–144. [IF2012 = 1.260] Carreras-Aubets M., Montero F.E., Kostadinova A., Carrasson M. (2012) Parasite communities in the red mullet, Mullus barbatus L., respond to small-scale variation in the levels of polychlorinated biphenyls in the Western Mediterranean. Marine Pollution Bulletin 64: 1853–1860. [IF2012 = 2.531] Carreras-Aubets M., Montero E.E., Kostadinova A., Gibson D.I., Carrasson M. (2012) Redescriptions of two frequently recorded but poorly known hemiurid digeneans, Lecithochirium musculus (Looss, 1907) (Lecithochiriinae) and Ectenurus lepidus Looss, 1907 (Dinurinae), based on material from the western Mediterranean. Systematic Parasitology 82: 185–199. [IF2012 = 1.260] Guilhem R., Šimková A., Morand S., Gourbiere S. (2012) Within-host competition and diversification of macroparasites. Journal of the Royal Society Interface 9(76): 2936–2946. [IF2012 = 5.658] Hernández-Orts J.S., Alama-Bermejo G., Carrillo J.M., García N.A., Crespo E.A., Raga J.A., Montero F.E. (2012) Aporocotyle mariachristinae n. sp., and A. ymakara Villalba & Fernandez, 1986 (Digenea: Aporocotylidae) of the pink cusk-eel Genypterus blacodes (Ophidiformes: Ophidiidae) from the Patagonia, Argentina. Parasite 19: 319–330. [IF2012 = 1.116] Hernández-Orts J.S., Alama-Bermejo G., Crespo E.A., García N.A., Raga J.A., Montero F.E. (2012) Breizacanthus aznari sp. n. (Acanthocephala: Arhythmacanthidae) from the banded cusk-eel Raneya brasiliensis (Ophidiiformes: Ophidiidae) from the Patagonian coast in Argentina. Folia Parasitologica 59: 264–271. [IF2012 = 2.515] Holzer A.S., Bartošová P., Pecková H., Tyml T., Atkinson S., Bartholomew J., Sipos D., Eszterbauer E., Dyková, I. (2012) ‘Who's who’ in renal sphaerosporids (Bivalvulida: Myxozoa) from common carp, Prussian carp and goldfish – molecular identification of cryptic species, blood stages and new members of Sphaerospora sensu stricto. Parasitology 24: 1–15. [IF2012 = 2.556] Kraľová-Hromadová I., Bazsalovicsová E., Oros M., Scholz T. (2012) Sequence structure and intragenomic variability of ribosomal ITS2 in monozoic tapeworms of the genus Khawia (Cestoda: Caryophyllidea), parasites of cyprinid fish. Parasitology Research 111: 1621–1627. [IF2012 = 2.852] Kuchta R., Burianová A., Jirků M., de Chambrier A., Oros M., Brabec J., Scholz T. (2012) Bothriocephalidean tapeworms (Cestoda) of freshwater fish in Africa, including erection of Kirstenella n. g. and description of Tetracampos martinae n. sp. Zootaxa 3309: 1–35. [IF2012 = 0.974] Mašová Š. (2012) Structure of the cephalic end and eggs of female Cithariniella khalili Petter, Vassiliadès et Troncy, 1972 (Nematoda: Pharyngodonidae), a parasite of African fishes. Helminthologia 49(2): 115–118. [IF2012 = 0.952] Mendlová M., Desdevises Y., Civáňová K., Pariselle A., Šimková A. (2012) Monogeneans of West African cichlid fish: evolution and cophylogenetic interactions. PLoS ONE 7(5): e37268. [IF2012 = 4.208] Mendoza-Palmero C.A., Scholz T., Mendoza-Franco E.F., Kuchta R. (2012) New species and new geographical records of dactylogyrids (Monogenea) of catfish (Siluriformes) from the Peruvian Amazonia. Journal of Parasitology 98: 484–497. [IF2012 = 1.321] Moravec F., Bilal S.J., Abdullah S.M.A. (2012) Two species of Rhabdochona (Nematoda: Rhabdochonidae) from the cyprinid fish Luciobarbus kersin (Heckel) in northern Iraq, including R. (Globochona) kurdistanensis sp. n. Folia Parasitologica 59: 139–147. [IF2012 = 2.515] Moravec F., Chavan S.P. (2012) Female morphology of Philometra hyderabadensis Rasheed, 1963 (Nematoda: Philometridae), a little-known parasite of the catfish Wallago attu Bloch & Schneider in India, as revealed by SEM observations. Systematic Parasitology 83: 117–122. [IF2012 = 1.260] Moravec F., Ermolenko A.V., Besprozvannykh V.V., Scholz T. (2012) New data on the morphology of some Far- Eastern species of Rhabdochona (Nematoda: Rhabdochonidae), as revealed by SEM observations. Folia Parasitologica 59: 195–208. [IF2012 = 2.515] 101

Moravec F., Jassim A.A.R., Al-Salim N.K. (2012) Philometroides acanthopagri sp. nov., a new philometrid (Nematoda, Philometridae) from the musculature of Acanthopagrus latus (Sparidae) from marine waters of Iraq. Acta Parasitologica 57: 372–377. [IF2012 = 1.000] Moravec F., Kamchoo K. (2012) Description of Rhabdochona (Globochona) rasborae sp. n. (Nematoda: Rhabdochonidae) from the freshwater cyprinid fish Rasbora paviana Tirant in southern Thailand. Folia Parasitologica 59: 209–215. [IF2012 = 2.515] Moravec F., Nagasawa K., Nohara K. (2012) Two species of philometrid nematodes (Philometridae) from marine fishes off Japan, including Philometroides branchiostegi sp. n. from Branchiostegus japonicus (Malacanthidae). Folia Parasitologica 59: 71–78. [IF2012 = 2.515] Moravec F., Taraschewski H., Appelhoff D., Weyl O. (2012) A new species of Hysterothylacium (Nematoda: Anisakidae) from the giant mottled eel Anguilla marmorata in South Africa. Helminthologia 49: 174–180. [IF2012 = 0.783] Moravec F., Walter T., Yuniar A.T. (2012) Five new species of philometrid nematodes (Philometridae) from marine fishes off Java, Indonesia. Folia Parasitologica 59: 115–130. [IF2012 = 2.515] Moravec F., Yost J., de Buron I. (2012) New data on the morphology of Iheringascaris inquies (Linton, 1901) (Nematoda: Anisakidae), a specific parasite of the marine fish Rachycentron canadum (Linnaeus), as revealed by SEM. Folia Parasitologica 59: 315–320. [IF2012 = 2.515] Ondračková M., Šimková A., Civáňová K., Jurajda P. (2012) Parasite diversity and microsatellite variability in native and introduced populations of Neogobius species (Gobiidae). Parasitology 139(11): 1493–1505. [IF2012 = 2.556] Oros M., Ash A., Brabec J., Kar P.K., Scholz T. (2012) A new monozoic tapeworm, Lobulovarium longiovatum n. g., n. sp. (Cestoda: Caryophyllidea), from barbs Puntius spp. (Teleostei: Cyprinidae) in the Indomalayan region. Systematic Parasitology 83: 1–13. [IF2012 = 1.260] Palíková M., Navrátil S., Dyková I., Pavlík I., Slaný M., Tichý F., Novotný L., Zendulková D., Kříž P., Laichmanová M., Mareš J. (2012) Archamoeba infection manifested by granulomatous inflammatory lesions in European tench, Tinca tinca (L.). Bulletin of the European Association of Fish Pathologists 32: 174−179. [IF2012 = 0.729] Přikrylová I., Blažek R., Vanhove M.P.M. (2012) An overview of the Gyrodactylus (Monogenea: Gyrodactylidae) species parasitizing African catfishes, and their morphological and molecular diversity. Parasitology Research 110: 1185–1200. [IF2012 = 2.852] Přikrylová I., Blažek R., Gelnar M. (2012) Gyrodactylus malalai sp. nov. (Monogenea: Gyrodactylidae) from Nile tilapia, Oreochromis niloticus (L.) and Redbelly tilapia, Tilapia zillii (Gervais) (Teleostei: Cichlidae) in the Lake Turkana, Kenya. Acta Parasitologica 57: 122–130. [IF2012 = 1.000] Přikrylová I, Vanhove M.P.M, Janssens S.B., Billeter P.A., Huyse T. (2012) Tiny worms from a mighty continent: high diversity and new phylogenetic lineages of African monogeneans. Molecular Phylogenetics and Evolution 67(1): 43–52. [IF2012 = 4.409] Seifertová M., Bryja J., Vyskočilová M., Martínková N., Šimková A. (2012) Multiple Pleistocene refugia and postglacial colonization in the European chub (Squalius cephalus) revealed by combined use of nuclear and mitochondrial markers. Journal of Biogeography 39: 1024–1040. [IF2012 = 5.078] Šimková A., Navrátilová P., Dávidová M., Ondračková M., Sinama M., Chappaz R., Gilles A., Costedoat C. (2012) Does invasive Chondrostoma nasus shift the parasite community structure of endemic Parachondrostoma toxostoma in sympatric zones? Parasites & Vectors 5(1): 200. [IF2012 = 3.724] Scholz T., de Chambrier A. (2012) A new genus and species of proteocephalidean tapeworm (Cestoda) from Pangasius larnaudii (Siluriformes: Pangasiidae) in South East Asia. Journal of Parasitology 98: 648–653. [IF2012 = 1.321] Vrtílek M., Reichard M. (2012) An indirect effect of biological invasions: the effect of zebra mussel fouling on parasitisation of unionid mussels by bitterling fish. Hydrobiologia 696: 205–214. [IF2012 = 2.212] Yoneva A., Levron C., Ash A., Scholz T. (2012) Spermatological characters of monozoic tapeworms (Cestoda: Caryophyllidea), including first data on a species from the Indomalayan catfish. Journal of Parasitology 98: 423–430. [IF2012 = 1.321] Yoneva A., Levron C., Nikolov P.N., Mizinska Y., Mariaux J., Georgiev B.B. (2012) Spermiogenesis and spermatozoon ultrastructure of the paruterinid cestode Notopentorchis sp. Parasitology Research 111: 135–142. [IF2012 = 2.852] Yoneva A., Levron C., Oros M., Orosová M., Scholz T. (2012) Spermiogenesis and spermatozoon ultrastructure of caryophyllidean cestode Hunterella nodulosa (Cestoda: Caryophyllaeidae). Folia Parasitologica 59: 179– 186. [IF2012 = 2.515]

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Books/book chapters

Atkinson S.D., Bartošová-Sojková P., Whipps C.M., Bartholomew J.L. (2015) Chapter 6: Approaches for characterizing myxozoan species. In: B. Okamura et al. (Eds.), Myxozoan Evolution, Ecology and Development. Switzerland, Cham, Springer International Publishing: pp. 111–123. de Chambrier A., Scholz T., Mariaux J., Kuchta R. (2016) Onchoproteocephalidea I. In: J.N. Caira and K. Jensen (Eds.), Tapeworms from vertebrate bowels of the earth 2008–2017. Special publication No. 25. Natural History Museum, The University of Kansas, Allen Press, Inc., Lawrence, Kansas: pp. 251–277. Durborow R., Kuchta R., Scholz T. (2015) Chapter 8. Paddlefish Diseases. In: Mims S.D., Shelton W.L. (Eds.), Paddlefishes Aquaculture. Wiley Blackwell, Hoboken, New Jersey, pp. 227–289. Dyková I., Kostka M. (2013) Illustrated guide to culture collection of free-living amoebae. Czech Republic, Praha, Academia: 363 pp. Janáč M., Reichard M. (2016) Use of drift nets to infer fish transport and migration strategies in inland aquatic ecosystems. In Morais P., Davrat F. (eds) An Introduction to Fish Migration, pp 197–213. CRC Press. Feist S.W., Morris D.J., Alama-Bermejo G., Holzer A.S. (2015) Chapter 8: Cellular processes. In: B. Okamura et al. (Eds.), Myxozoan Evolution, Ecology and Development. Switzerland, Cham, Springer International Publishing: pp. 139–154. Faltýnková A., Georgieva S., Kostadinova A., Bray R.A. (2017) Biodiversity and evolution of digeneans of fishes in the Southern Ocean. In: H. Mehlhorn and S. Klimpel (Eds.): Biodiversity and Evolution of Parasitic Life in the Southern Ocean. Springer, Parasitology Research Monographs, pp. 49–75. Fiala I., Bartošová-Sojková P., Okamura B., Hartikainen H. (2015) Chapter 4: Adaptive radiation and evolution within the Myxozoa. In: B. Okamura et al. (Eds.), Myxozoan Evolution, Ecology and Development. Springer International Publishing, Cham, Switzerland, pp. 69–84. Fiala I., Bartošová-Sojková P., Whipps C.M. (2015) Chapter 5: Classification and phylogenetics of Myxozoa. In: B. Okamura et al. (Eds.), Myxozoan Evolution, Ecology and Development. Springer International Publishing, Cham, Switzerland, pp. 85–110. Kuchta R., Scholz T. (2017) Bothriocephalidea. In: J.N. Caira and K. Jensen (Eds.), pp. 29–45. Kuchta R., Scholz T. (2017) Diphyllobothriidea. In: J.N. Caira and K. Jensen (Eds.), pp. 167–189. Kuchta R., Scholz T. (2017) Haplobothriidea. In: J.N. Caira and K. Jensen (Eds.), pp. 201–206. Kuchta R., Scholz T. (2017) Spathebothriidea. In: J.N. Caira and K. Jensen (Eds.), pp. 349–356. Kuchta R., Scholz T., Hansen H. (2017) Gyrocotylidea. In: J.N. Caira and K. Jensen (Eds.), pp. 191–199. Moravec F. (2013) Parasitic Nematodes of Freshwater Fishes of Europe. Revised Second Edition. Czech Republic, Praha, Academia: 601 pp. Passos C., Tassino B., Rosenthal G.G., Reichard M. (2015) Reproductive behavior and sexual selection in annual fishes. In: Berois, N., García, G., de Sá, R. (Eds.), Annual Fishes: Life History Strategy, Diversity, and Evolution. CRC Press. (207–230) 327 pp. Scholz T., Vanhove M.P.M., Smit N., Jayasundera Z., Gelnar M. (2018) A guide to the parasites of African freshwater fishes. Abc Taxa 18: 417 pp. Scholz T., Brabec J., Kuchta R. (2017) Nippotaeniidea. In: J.N. Caira and K. Jensen (Eds.), pp. 243–250. Scholz T., Kuchta R. (2017) Amphilinidea. In: J.N. Caira and K. Jensen (Eds.), pp. 21–28. Scholz T., Oros M. (2017) Caryophyllidea. In: J.N. Caira and K. Jensen (Eds.), pp. 47–64. Šimková A., Morand S. (2014) Parasite species coexistence and the evolution of the parasite niche. In: Morand S., Littlewood T. and Krasnov B. (eds.) Parasite Diversity and Diversification - Evolutionary Ecology Meets Phylogenetics. Cambridge University Press, Cambridge, UK, pp. 360–375. Šimková A., Rohde K. (2013). Community stability and instability in ectoparasites of marine and freshwater fish. In: Rohde K. (ed.). Balance of Nature and Human Impact. Cambridge University Press, Cambridge, UK, pp. 75– 87.

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7th Workshop of European Centre of Ichthyoparasitology 28 November 2018 Programme & Abstracts

Michal Benovics, Zuzana Jayasundera & Milan Gelnar (Eds.)

Graphic design of cover: Zuzana Jayasundera, Oliver Staša Autor of the cover photo: R. Blažek, A. de Chambrier and R. Kuchta Technical redactor: Michal Benovics, David Baláš Published by Masaryk University with financial support ECIP; centre of the Czech Science Foundation; project No. P505/12/G112. Brno 2018, 1st edition

ISBN 978-80-210-9079-8 ISBN 978-80-210-9083-5 (online: pdf)

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