Iveta Matějusová

Phylogenetic relationships of the Dactylogyridae Bychowsky, 1933 (Monogenea: Dactylogyridea): the need for the systematic revision of the Ancyrocephalinae Bychowsky, 1937

Phylogenetic analyses based on partial 18S rDNA sequences of polyonchoinean monogeneans were conducted in order to investigate the relationships between selected families and subfamilies of the Dactylogyrinea, mainly within the Dactylogyridae. We tested the status of the Ancyrocephalidae sensu Bychowsky & Nagibina (1978) and the Ancyrocephalinae sensu Kritsky & Boeger (1989). Within the Dactylogyrinea, the Diplectanidae and Dactylogyridae are well supported by maximum likelihood and maximum parsimony analyses, but their phylogenetic relationship with the Pseudomurraytrematidae remains unresolved. Phylogenetic relationships between the Pseudodactylogyrinae, Ancyrocephalinae, Ancylodiscoidinae and Dactylogyrinae indicate paraphyly of the Ancyrocephalidae sensu Bychowsky & Nagibina (1978). The group of species recently considered as the Dactylogyridae sensu Kritsky & Boeger (1989) comprises two sister groups. The first group includes the freshwater Ancyrocephalinae and the Ancylodiscoidinae. The second group includes the Pseudodactylogyrinae, Dactylogyrinae and the Ancyrocephalinae from the fish species Siganus doliatus and Tetraodon fluviatilis. The non-monophyly of the Ancyrocephalinae (i.e. the non-monophyly of the group of species recently considered as members of Ancyrocephalinae), previously suggested by Kritsky & Boeger (1989) using the morphological characters, indicates that classification of the Dactylogyridae needs to be revised.

Seasonal occurrence and metrical variability of Gyrodactylus rhodei Žitňan 1964 (Monogenea, Gyrodactylidae)

The seasonal dynamics of Gyrodactylus rhodei, a monogenean ectoparasite of bitterling (Rhodeus sericeus), was studied from June 2000 to May 2001 in the Kyjovka River, Czech Republic. A negative relationship between prevalence and intensity of infection of G. rhodei and water temperature was found. Metrical variability of the hard parts of the parasite haptor was studied throughout the sampling season. A negative relationship between water temperature and the size of the hard parts of the G. rhodei haptor was evident in the measurements of the total length of the marginal hooks, the sickle length of marginal hooks, anchors, anchor point and root, the width of the ventral bar and the membrane processes. Sequences of the partial ITS (rDNA) of specimens collected during the cold and warm seasons were analysed. Sequences of all studied parasite specimens were identical and there was no evidence of intraspecific variability in the sequenced region.

Nestedness in assemblages of gyrodactylids (Monogenea: Gyrodactylidea) parasitising two species of cyprinid – with reference to generalists and specialists

The structure of gyrodactylid assemblages in individual fishes of two species of cyprinid was determined. A total of 100 specimens of minnow, Phoxinus phoxinus, and 137 specimens of roach, Rutilus rutilus, were investigated for presence of gyrodactylids. Host specificity, specialists vs. generalists, was noted in each host fish. A nested pattern was recorded in parasite assemblages of minnow, the host with a dominant number of specialist gyrodactylids. A non-nested pattern was observed in parasite assemblages of roach, the host with a dominant number of generalist gyrodactylids. The host specificity appears to be a meaningful factor that determines the pattern of gyrodactylid assemblages of both fish hosts.

Morphometric and molecular characterization of Parapharyngodon echinatus (Nematoda, Pharyngodonidae) from the Senegal gecko ( Tarentola parvicarinata )

A West African species of gecko, Tarentola parvicarinata Joger (Gekkonidae), collected in the Niokolo Koba National Park in East Senegal is a new host species for the nematode Parapharyngodon echinatus (Rudolphi, 1819). Fifty one specimens of P. echinatus were studied under a light microscope and thirteen specimens were studied for the first time by a scanning electron microscope. The main differences between P. echinatus and other African species, especially Parapharyngodon micipsae (Seurat, 1917), were in the shape of annules on body cuticle, broad lateral alae and their terminations, the shape of the distal extremity of the spicule and number of outgrowths at the anterior cloacal lip. For the purposes of DNA characterization, partial sequence of the small subunit ribosomal RNA gene (SSU rRNA) was obtained from two specimens of P. echinatus. This is the first published sequence of a species from the family Pharyngodonidae.

A new gyrodactylid (Monogenea) genus on gray bichir, Polypterus senegalus (Polypteridae) from Senegal (West Africa).

Abstract Diplogyrodactylus martini n. g. et sp., a viviparous monogenean, is described from the gills of gray bichir Polypterus senegalus Cuvier, 1829, in Senegal. This new genus can be readily distinguished from the other described viviparous genera based on the morphology of the attachment apparatus. The haptor is equipped with a pair of centrally positioned hamuli, a simple ventral bar without membrane and 8 pairs of marginal hooks of 2 types, 5 pairs of hooks with well-defined large falculate sickles, and 3 pairs of smaller hooks with well-articulated sickles. The new genus is unique in having a teardrop-shaped muscular tubular male copulatory organ which lacks spines. A partial sequence of the internal transcribed spacer (ITS rDNA) was obtained from 4 specimens of D. martini. The relationship among D. martini and other genera of the Gyrodactylidae is discussed based on the similarities of the attachment apparatus.

Morphometric comparison of three members of the Gyrodactylus nemachili-like species group (Monogenea: Gyrodactylidae) on Barbatula barbatula L. in the Czech Republic, with a reinstatement of G. papernai Ergens & Bychowsky, 1967

The Gyrodactylus nemachili-like species complex, parasitising stone loach Barbatula barbatula L. in the Czech Republic, was investigated based on the detailed morphometric analysis of its haptoral hard parts. The existence of two species, G. jiroveci Ergens & Bychowsky, 1967 and G. pseudonemachili Ergens & Bychowsky, 1967 was confirmed. A standard discriminant analysis of the morphometric data showed a clear separation between specimens of G. jiroveci and another species or variant. Based on the morphology of the haptoral hard parts, this third species was more similar to G. jiroveci than to G. pseudonemachili. The differences between G. jiroveci and the third species were mainly in the morphology of the marginal hook sickle, but included the total length of the marginal hook, the length of the hamulus point, the length of the marginal hook sickle, the width of the ventral bar, and the width of the dorsal bar. Based on comparison with the original material, the third species was identified as G. papernai Ergens & Bychowsky, 1967, which has recently been considered a synonym of G. jiroveci. These analyses provided sufficient evidence for the restoration of G. papernai as a valid taxon. Partial sequences of the internal transcribed spacer rRNA genes (ITS rDNA) verified the validity of all three Gyrodactylus species studied.

Ethanol-fixed material used for both classical and molecular identification purposes: Eudiplozoon nipponicum (Monogenea: Diplozoidae) as a case parasite species

This study is focused on the feasibility of two treatments of alcohol-fixed monogenean parasites which are intended to be use for the combined morphological and molecular characterizations. The monogenean parasite, Eudiplozoon nipponicum, was selected as a model parasite species; however it is expected that these techniques will be suitable for other monogeneans and other parasitic families. The haptor of diplozoid parasites is equipped with sclerotized attachment clamps and central hooks which are utilized for morphological identification. As parasite tissue become very tough and rigid when preserved in ethanol, using these structures for species identification without additional treatment is difficult. We investigated two different techniques to digest the surrounding tissues, the first was treatment with 10% sodium dodecyl sulphate (SDS) and the second treatment was proteinase K. Tissue was successfully digested in both treatments and all clamps, central hook and even individual sclerites of the clamps were clearly visible and well defined. After treatment, the digest was used to extract genomic DNA, and the second internal transcribed spacer of the ribosomal DNA genes (rDNA) was amplified. Nucleic acid sequence was obtained from 90% of parasite specimens processed by both treatments. Treatment of haptors with SDS was proven to be more successful with no visible changes or damage observed to sclerites even after a month. This method represents a useful tool for the combined morphological and molecular studies as the correct sequence can be assigned to the same individual worm from which haptoral parts have been obtained.

Thelastoma gueyei sp. n. (Nematoda: Thelastomatidae) from the Senegal diplopod Archispirostreptus tumuliporus (Diplopoda: Spirostreptidae)

Thelastoma gueyei sp. n., a nematode belonging to the long-tailed species group of the genus Thelastoma, is described from the diplopod Archispirostreptus tumuliporus (Spirostreptidae) collected in Niokolo Koba National Park (Senegal, West Africa).Thelastoma gueyei sp. n. is morphologically most similar to T. gipetiti. Females are characterised by: the vulva being situated in theposterior half of the body and near to the anus (V_ = 75-86) with the anterior vulval lip developed into a prominent flap; excretory pore located at the level of the anterior end of the pharyngeal bulb; b_ = 28-38 and tail, expressed as a proportion of L_, = 1.7-2.7. Males have narrow cuticular alae extending from about the middle of the pharynx to the level of the anteriormost pair of copulatorypapillae; four pairs of copulatory papillae, two large, subventral, pairs being located adcloacally on the genital cone, a third, much smaller pair on the posterior margin of the genital cone and the last pair being situated at the mid-point of the tail spike; and a tailoccupying 10.8-13.2% of the total body length. The distal tip of the spicule is drop-shaped. Morphological characteristics were studied using scanning electron microscopy and a comparison of the long-tailed group of thelastomatids is provided. Nucleic acid sequence ofthe small subunit ribosomal RNA gene was obtained for purposes of DNA barcoding.

Octomacrum europaeum (Monogenea: Octomacridae) in Europe: historical and recent perspectives

Abstract. Several specimens of Octomacrum europaeum (Octomacridae: Monogenea) were found on the gills of Alburnoides bipunctatus in the Czech Republic. The morphometry of the specimens measured in the present study was in agreement with that of the original description.

Modified sample preparation from ethanol fixed material for identification of Eudiplozoon nipponicum as a case parasite species for the combined morphological and molecular characterization

This study is focused on feasibility of two treatments of alcohol-fixed monogenean parasites that are intended to be used for the combined - molecular and morphological characterization. Conclusions of particular identification approaches highly correspond and could be used in subsequent phylogenetic studies.