Marek S. Ziętara

Identification of Gyrodactylus ectoparasites in Polish salmonid farms by PCR-RFLP of the nuclear ITS segment of ribosomal DNA (Monogenea, Gyrodactylidae)

The Gyrodactylus fauna of 274 fish taken from ten salmonid farms in Poland was sampled in 2006. Four fish species were investigated: rainbow trout Oncorhynchus mykiss, brown trout Salmo trutta (morphs fario, lacustris, and trutta), grayling Thymallus thymallus and huchen Hucho hucho. No parasites were observed on huchen. No indications of gyrodactylosis were observed, but an unexpected parasite species diversity was found. A molecular species identification by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of ITS1 + 5.8S + ITS2 was utilized, with addition of morphometric methods. The most frequent parasite was a new record in Poland, G. teuchis. It was present in two molecular forms on brown trout and rainbow trout, which also carried G. derjavinoides and G. truttae. Three molecular forms of G. salaris/G. thymalli were found, the standard type ITS only on grayling. A heterozygous (or heterogenic) G. salaris type described earlier in Denmark was found in seven farms on rainbow trout, and a complementary homozygous clone which differs from the standard by three nucleotides, in two farms. This homozygous form has not been recorded earlier. The PCR-RFLP results were confirmed by sequencing ITS segment from representative specimens of each type and comparing them with all available salmonid-specific Gyrodactylus sequences in GenBank. The Polish fauna with seven different Gyrodactylus clones separated by PCR-RFLP was the most diverse reported in fish farms in any country so far.

TWO NEW ANTARCTIC GYRODACTYLUS SPECIES (MONOGENOIDEA): DESCRIPTION AND PHYLOGENETIC CHARACTERIZATION

Abstract Two new species of Gyrodactylus Nordmann, 1832 (Gyrodactylidae, Monogenoidea) are described, i.e., Gyrodactylus coriicepsi n. sp. from gills of black rockcod, Notothenia coriiceps Richardson, 1844, and Gyrodactylus nudifronsi n. sp. from gills of gaudy notothen, Lepidonotothen nudifrons Lönnberg, 1905, collected in waters surrounding the South Shetland Islands, Antarctica. Gyrodactylus coriicepsi n. sp. is characterized by a long, narrowing membrane of the ventral bar and a marginal hook sickle with beveled heel and toe. Gyrodactylus nudifronsi n. sp. is characterized by a conspicuous ventral bar process and barely curved marginal hook sickle with curved heel and toe. Definitive species identification and phylogenetic position were based on sequences of the rDNA region (ITS1-5.8S-ITS2). Both new species belong to a monophyletic group (bootstrap support 99%) that includes the European marine species Gyrodactylus perlucidus Bykhovskiy and Polyanskiy, 1953 (on Zoarces viviparus), Gyrodactylus flesi Malmberg, 1957 and G. robustus Malmberg, 1957 (both on Platichthys flesus), and the freshwater species Gyrodactylus hrabei Ergens, 1957 and Gyrodactylus mariannae Winger, Hansen, Bachmann and Bakke, 2008 (both on Cottus poecilopus). The observations suggest an evolutionary continuum of the marine Gyrodactylus spp. fauna in the northern and southern hemispheres, which indicates the marine origin of some freshwater Gyrodactylus species.

A monogenean fish parasite, Gyrodactylus chileani n. sp., belonging to a novel marine species lineage found in the South-Eastern Pacific and the Mediterranean and North Seas

Gyrodactyluschileani n. sp. is the first Gyrodactylus species reported from Chile. It is an ectoparasite living on fins and skin of a small fish, the Chilean tidal pond dweller Helcogrammoides chilensis (Cancino) (Perciformes: Tripterygiidae). A phylogenetic analysis based on 5.8S+ITS2 of rDNA placed the new species close to marine Gyrodactylus species found in Europe: G. orecchiae Paladini, Cable, Fioravanti, Faria, Cave & Shinn, 2009 on gilthead seabream Sparus aurata L. from the Adriatic and Tyrrhenian Sea fish farms (Perciformes: Sparidae), and an undescribed species on the black goby Gobius niger L. from the North Sea (Perciformes: Gobiidae). A morphological description of the latter species is unavailable. These geographically distant parasite samples on different host families form a new well supported Gyrodactylus orecchiae lineage. Using molecular phylogenetics, it is shown that the marine species groups of Gyrodactylus may have a worldwide distribution.

Morphological and molecular characterization of Ceratomyxa gurnardi sp. n. (Myxozoa: Ceratomyxidae) infecting the gallbladder of the grey gurnard Eutrigla gurnardus (L.) (Scorpaeniformes, Triglidae)

The myxosporean specimens were noted in grey gurnard Eutrigla gurnardus (L.) from the area near the Shetland Islands. The structure and dimensions of its vegetative stage differ from earlier descriptions. A sequence of small subunit ribosomal RNA gene obtained during the current study differs from other Ceratomyxa spp. available in GenBank. A phylogenetic position of parasite based on the 18S rDNA fragment was estimated. The proposed name for this myxosporean is Ceratomyxa gurnardi sp. n.

Horizontal transmission of the ectoparasite Gyrodactylus arcuatus (Monogenea: Gyrodactylidae) to the next generation of the three-spined stickleback Gasterosteus aculeatus

In the parthenogenetic monogeneans of the genus Gyrodactylus Nordmann, 1832, the genetic diversity within or between hosts is determined by the relative roles of lateral transmission and clonal propagation. Clonality and limited transmission lead to high-amplitude metapopulation dynamics and strong genetic drift. In Baltic populations of the three-spined stickleback Gasterosteus aculeatus Linnaeus, the local mitochondrial diversity of Gyrodactylus arcuatus Bychowsky, 1933 is very high, and spatial differentiation weak. To understand the transmission dynamics in a single location, the transmission of the parasite from adults to next generation sticklebacks was investigated in a northern Baltic brackish water location. By sequencing 777 nt of cox1, as many as 38 separate mitochondrial haplotypes were identified. In August, the intensity of gyrodactylid infection on adult hosts was high, the haplotype diversity (h) was extreme and differentiation between fish was negligible (total h = 0.926, mean h = 0.938). In October, only 46% of the juvenile sticklebacks carried G. arcuatus. The number of parasites per young fish followed a Poisson distribution 0.92 ± 1.04 (mean ± SD) on October 2, and was clearly overdispersed 2.38 ± 5.00 on October 25. The total haplotype diversity of parasites on juveniles was nearly as high as in adults (h = 0.916), but the mean per fish was only h = 0.364 (FST = 0.60), due to low intensity of infection and rapid clonal propagation of early arrivals. The initial first come first served advantage of the first gyrodactylid colonisers will be lost during the host adulthood via continuous transmission. Nesting and polygamy are suggested as factors maintaining the high genetic diversity of the parasite population. The transmission dynamics and, consequently, the population structure of Baltic G. arcuatus is fundamentally different from that of G. salaris Malmberg, 1957, on the Baltic salmon Salmo salar Linnaeus.

Revision of Gyrodactylus salaris phylogeny inspired by new evidence for Eemian crossing between lineages living on grayling in Baltic and White sea basins

In this research, grayling-specific Gyrodactylus salaris Malmberg, 1957 isolates from Baltic Sea basin were collected in Sweden for the first time. Samples were obtained in three drainage systems: Kalixälven (River Kaitum), Ljungan (River Sölvbacka strömmar), and Umeälven (River Juktån). Three molecular markers were analysed: nuclear ITS rDNA (Internal Transcribed Spacer) and ADNAM1 (Anonymous DNA Marker 1), and mitochondrial cox1 gene. As a result, four new mitochondrial haplotypes were identified (III-C1tt, III-C1ttht, IX-A1tt and X-A1tt). The ADNAM1 analyses resulted in revealing two new alleles (WS4 and BS9) and two new genotypes (T6 and T7). T7 seems to be an indicator of ancient crossing between Baltic and White Sea lineages of the parasite which happened during a first 3000-year period of Eemian interglacial about 130,000 years ago in the connection between Baltic and White Sea. Molecular clock estimates were adjusted, revealing the mean substitution rate and the divergence rate among branches of 3.6% (95% HPD: 2.2%–5.2%) and 7.2% per million years, respectively. As a result, cox1 phylogeny rooted with the introgressed haplotypes has been revised and altered in accordance to new data, revealing fourteen equidistant lineages five of which have been excluded from the study. Based on the new phylogenetic approach, including the molecular clock, this work suggests an overall revision of G. salaris phylogeny and attempts at precisely drawing the division of lineages within this polytypic species as well as proposes unification in nomenclature for its strains.