First report of Gyrodactylus spp. (Platyhelminthes: Monogenea) in the western Mediterranean Sea: molecular and morphological descriptions

The Gyrodactylus spp. fauna on species of gobies, Pomatoschistus, Gobiusculus, and Knipowitschia (Gobiidae: Teleostei), from the western Mediterranean and Adriatic seas is strikingly similar to that found in the Baltic Sea and eastern Atlantic Ocean, both in morphology and in internal transcribed spacer (ITS) rRNA. The fauna consisted of Gyrodactylus branchialis, G. ostendicus, G. gondae, G. rugiensis, G. rugiensoides, and G. arcuatus. No new species have been found. A morphometric comparison between G. branchialis from the Mediterranean and Adriatic seas and its type locality in Ostend (Belgium) showed significant differences in ventral bar and marginal hook features. The morphometric variation was lower in G. rugiensis, whereas no significant differences were found in G. ostendicus. Gyrodactylus branchialis and G. ostendicus collected on P. microps were slightly different in the ITS rDNA (-0.6%) compared with specimens on the closely related P. marmoratus, probably reflecting ongoing speciation. A hybrid zone was identified in the Vaccarès lagoon complex (France) where both host species are sympatric. There was no clear geographic or host-related pattern in the variation found in the ITS2 rDNA in G. arcuatus sampled from P. microps, G. flavescens, Pungitius pungitius, K. panizzae, and its original host Gasterosteus aculeatus (2 polymorphic sites). Of all studied species, only G. arcuatus, G. rugiensis, and G. rugiensoides showed minor intraspecific variation in the ITS rDNA. Hence, the physical separation by a shoreline of more than 10,000 km is hardly reflected in the parasite ITS rDNA.     

An electrophoretic study of the systematic relationships of some closely related goby species (Pisces, Gobiidae)

An electrophoretic study of genetic variation at 31 loci in each of nine closely related gobiid species was carried out in order to assess their systematic relationships. The species used were: Pomatoschistus canestrinii, P. lozanoi, P. marmoratus, P. microps, P. minutus, P. norvegkus, P. pictus, Knipowitschia caucasica, Neogobius melanostomus. Genetic distances ( D ) and identities ( I ) were calculated by Nei's method for all 36 pairwise comparisons and dendrograms were constructed.
The results show that the very similar and cross-fertile species pair, P. lozanoi and P. minutus , have the highest I value; P. norvegkus has high I values with both species. Pomatoschistus marmoratus and P. microps also display high genetic similarity with each other. The remaining Pomatoschistus species and Knipowitschia caucasica are broadly and moderately equidistant from each other and from the five species refferred to above, while Neogobius melanostomus is considerably more distant from all other species.
These findings essentially agree with the pattern of relationships established using classical taxonomic approaches. However, the observation that Knipowitschia caucasica is as similar to Pomatoschistus spp. as the latter are to each other suggests that the generic distinction of Knipowitschia could usefully be reconsidered.     

Four new species of Gyrodactylus von Nordmann, 1832 (Monogenea,Gyrodactylidae) on gobiid fishes: combined DNA and morphological analyses

Four Gyrodactylus species parasitising four closely related gobiid species in European coastal waters were studied and compared with G. arcuatus Bychowsky sensu Bychowsky & Poljansky (1953) from Gasterosteus aculatus . These were G. gondae n. sp. from Pomatoschistus minutus and P. lozanoi , G. flavescensis n. sp. from Gobiusculus flavescens , G. arcuatoides n. sp. from P. minutus and G. branchialis n. sp. from P. microps. Combined molecular and morphological analyses, as well as morphometric and statistical methods, were used. The ssrRNA V4 region and the complete ITS rDNA region were sequenced. Genetically the four new species are clearly distinct from G. arcuatus . From a morphological point of view, the haptoral hard parts of G. gondae n. sp., G. flavescensis n. sp. and G. arcuatoides n. sp. are related to those of G. arcuatus, while these parts of G. branchialis n. sp. are different, but related to those of G. quadratidigitus Longshaw, Pursglove & Shinn, 2003. For the latter two species, a new species group is formed. The V4 and ITS sequence analyses, however, indicate a close relationship between G. branchialis and the three G. arcuatus-like species.     

Dynamics of Kudoa camarguensis (myxosporean) infection in two gobiid species,Pomatoschistus microps and P. minutus (Teleostei: Pisces), in the Rhĵne River delta,France

The occurrence of the myxosporean parasite Kudoa camarguensis was surveyed monthly during 1997 in a brackish-water lagoon of the Rh?ne River delta (France). K. camarguensis was found on its typical host, Pomatoschistus microps, and on an additional host, P. minutus. Prevalence and mean abundance were higher in the typical host than in the additional host due to differences in the temporal occupancy of the lagoon by the 2 species. The temporal occurrence of this myxosporean parasite is discussed in relation to the migratory habits of P. minutus and the sedentary habits of P. microps.     

The food of Pomatoschistus minutus (Pisces, Gobiidae) in Belgian coastal waters, and a comparison with the food of its potential competitor P. lozanoi

Data are presented on the food and gonadosomatic index of Pomatoschistus minutus obtained from the bycatch of a commercial shrimp trawler. P. minutus has a diverse diet including zooplankton and hyperbenthos but mainly epibenthic and benthic animals. During the spawning season males consumed high numbers of Pomatoschistus eggs, presumably after aggressive take–overs of territories. The food of P. minutus is compared to the food of P. lozanoi of the same size classes obtained from the same hauls. In the absence of data on predator and prey abundance the discussion on possible interference competition between the two species remains largely speculative.     

Patterns of intermediate host use and levels of association between two conflicting manipulative parasites.

For many parasites with complex life cycles, manipulation of intermediate host phenotypes is often regarded as an adaptation to increase the probability of successful transmission. This phenomenon creates opportunities for either synergistic or conflicting interests between different parasite species sharing the same intermediate host. When more than one manipulative parasite infect the same intermediate host, but differ in their definitive host, selection should favour the establishment of a negative association between these manipulators. Both Polymorphus minutus and Pomphorhynchus laevis exploit the amphipod Gammarus pulex as intermediate host but differ markedly in their final host, a fish for P. laevis and a bird for P. minutus. The pattern of host use by these two conflicting manipulative parasites was studied. Their incidence and intensity of infection and their distribution among G. pulex were first examined by analysing three large samples of gammarids collected from the river Tille, Eastern France. Both parasites had low prevalence in the host population. However, temporal fluctuation in the level of parasitic infection was observed. Overall, prevalence of both parasite species was higher in male than in female G. pulex. We then assessed the degree of association between the two parasites among their intermediate hosts, using two different methods: a host-centred measure and a parasite-centred measure. Both measures gave similar results; showing random association between the two acanthocephalan species in their intermediate hosts. We discuss our results in relation to the selective forces and ecological constraints that may determine the pattern of association between conflicting manipulative parasites.     

Fish communities along an oxygen-poor salinity gradient (Zeeschelde Estuary, Belgium)

Fish living in the tidal fresh and brackish water reaches of the Zeeschelde Estuary were studied in samples collected from the cooling-water inlets of three power stations. Between July 1994 and June 1995, 42 different species were recorded including 26 marine migrants, 14 freshwater species and two diadromous species. Species number as well as fish abundance were correlated significantly with salinity and oxygen concentration. The community structure was analysed with a correlation biplot based on principal component analysis of the root-root transformed fish abundance. Four separated communities could be distinguished since the first principal component expressed a salinity and the second a temperature gradient. During summer and fall Pomatoschistus microps, P. minutus, P. lozanoi and Syngnathus rostellatus were most abundant in the brackish-water reach. At this time, freshwater species seemed to avoid the freshwater area. During winter, Sprattus sprattus, Clupea harengus and Dicentrarchus labrax were the dominant species of the brackish-water zone while the freshwater reaches were dominated by Gasterosteus aculeatus . Migrating fish such as Pleuronectes flesus, Lampetra fluviatilis, Anguilla anguilla and Osmerus eperlanus were restricted to the brackish environment.     

Parasites as indicator of fish behaviour

In the present study two species of gobies, Pomatoschistus microps and P. minutus from southwest Wales were studied and appearance of some unusual parasite of gobies, were used as an indicator for fish behaviour. The occurrence of pleurocercoids of Ligula intestinalis in the body cavity of these fish and Proteocephalus sp., excysted metacercariae and adult of Timoniella spp., excysted metacercariae of Bucephalus baeri , the excysted metacercariae of Podocotyle atomon and progenetic adult of Bucephaloides gracilescens in their gut indicate host anadromous, cannibalistic and scavenging behaviour.     

Gyrodactylus longidactylus n. sp., a monogenean from Pomatoschistus lozanoi (de Buen) (Gobiidae) from the North Sea

Gyrodactylus longidactylus n. sp., a gyrodactylid monogenean parasitising the gills of Pomatoschistus lozanoi (de Buen, 1923) in the North Sea is described. It is a species with rather small anchors which are only connected to a ventral bar which lacks a ventral bar membrane. The marginal hooks have long handles, which are always longer than the total length of the anchor. The pharynx has long pharyngeal processes. Measurements, drawings of the opisthaptoral hard parts, penis and pharynx, and SEM micrographs of this monogenean are presented. G. longidactylus n. sp. is the first monogenean species described from P. lozanoi.     

Choice matters: incipient speciation in Gyrodactylus corydori (Monogenoidea: Gyrodactylidae)

We investigated how Gyrodactylus corydoriBueno-Silva and Boeger, 2009 exploits two sympatric host species, Corydoras paleatus (Jenyns, 1842) and Corydoras ehrhardti Steindachner, 1910. Specimens of G. corydori were collected from the Piraquara and Miringuava Rivers, State of Paraná, Brazil, between 2005 and 2006. A total of 167 parasites was measured from both host species. Nine morphometric features of the haptoral sclerites were measured and analyzed by discriminant analysis, cluster analysis and multivariate analysis of variance. A fragment of the mitochondrial cytochrome oxidase I gene (COI) (∼740 bp) and the rDNA internal transcribed spacers (ITS) (∼1200 bp) of G. corydori were sequenced. Bayesian and parsimony analyses of COI recognized two genetically structured clades of G. corydori, which corresponded closely with the two species of Corydoras. Twenty-eight haplotypes were detected (18 were exclusive to C. ehrhardti and seven were exclusive to C. paleatus). The same general pattern between parasites and host species was observed in the morphometric analyses. Nevertheless, poor correlation of genetic and morphometric variation strongly supports the plastic nature of the morphological variation of haptoral sclerites. The existence of two clades with limited gene flow would suggest that G. corydori already represents two cryptic species. However, the morphometric and molecular data showed that there is insufficient evidence to support two valid species. The low COI (0.1-6.2%) and ITS (0.09-3.5%) divergence within G. corydori suggest a recent separation of the lineages between distinct host species (less than 1 million years). As the hypothesis of secondary contact of the parasite demographic history was rejected, our results point to the possibility of sympatric incipient ongoing speciation of G. corydori to form distinct parasite lineages adapted to C. ehrhardti and C. paleatus. This may be a common event within the Gyrodactylidae, adding a yet unreported mode of adaptive speciation that helps to understand its rate of diversification.     

Age determination in Pomatoschistus mi nut us (Pallas) and Pomatoschistus microps (Kreyer) (Pisces: Gobiidae) from the upper Tagus estuary, Portugal

The use of scale inspection as a means of age-determination in Pomatoschistus microps and Pomatoschistus minutus in the Tagus estuary is confirmed, but annulus formation pattern is different from that described for P. microps in the area of the British Isles (Miller, 1975). In Britain, two annuli are laid down on the scales each year, but in the Tagus, only one annulus is formed. One possible reason for this difference is the fact that in the Tagus the peak breeding season and the lowest water temperatures coincide. Therefore, the two causes of annulus formation coexist at the same time of year.
Population structure in both species is dominated by the 0-group, which appears earlier (April/May) than reported for other areas. Maximum age seems not to exceed 26 months in P. microps and 32 months in P. minutus .     

A redescription of Pomatoschistm norvegicus (Collett, 1903) (Teleostei: Gobioidei) based on syntype material

A deeper water species of the Mediterranean-Atlantic gobiid genus Pomatoschistus, P. norvegicus (Collett, 1903), is redescribed from syntype material, including details of the lateral-line system and a summary of geographical distribution. The systematic status of P. norvegicus is briefly discussed with reference to the other nominal species, P. minutus (Pallas) and P. lozanoi (De Buen), of the 'P. minutus' complex.     

Occurrence of vertebral deformities in Gobiidae (Pisces) from the Tagus estuary

In four species of gobiid fish caught in Tagus estuary during 1996, the incidence of skeletal deformities ranged from 2 to 5%. Differences between sampling stations or between the two most abundant species Pomatoschistus minutus and Gobius niger were statistically not significant (p > 0.05). The type of deformity depended upon species. P. minutus and P. microps showed bone nodules at the inferior side of the abdominal vertebrae, whilst G. niger and G. paganellus had shortened caudal vertebrae (achondroplasia) and deformed ural plates. The possible role of pollution or other conditions in the Tagus estuary is discussed.     

Experimental infections of the monogenean Gyrodactylus turnbulli indicate that it is not a strict specialist

Parasites represent a threat to endangered fish species, particularly when the parasite can host switch and the new host is vulnerable. If the parasite is highly host specific then successful host switching should be a rare occurrence; however, the host range of many parasites which are assumed to be specialists has never been tested. This includes the monogenean Gyrodactylus turnbulli, a well-studied ectoparasite found caudally on its known host, the guppy, Poecilia reticulata. In this study, we monitored parasite establishment and reproduction on a range of poeciliids and more distantly related fish. Individually maintained fish were experimentally infected with a single parasite and monitored daily to establish whether G. turnbulli could survive and reproduce on other fish species. Gyrodactylus turnbulli can infect a wider range of hosts than previously considered, highlighting the fact that host specificity can never be assumed unless experimentally tested. Our findings also have significant implications for parasite transmission to novel hosts and provide further insight into the evolutionary origins of this ubiquitous group of fish pathogens. Previous molecular evidence indicates that host switching is the key mechanism for speciation within the genus Gyrodactylus. Until recently, most Gyrodactylus spp. were assumed to be narrowly host specific. However, our findings suggest that even so-called specialist species, such as G. turnbulli, may represent a threat to vulnerable fish stocks. In view of the potential importance of host switching under artificial conditions, we propose to describe this as 'artificial ecological transfer' as opposed to 'natural ecological transfer', host switching under natural conditions.     

Small-scale clinal variation, genetic diversity and environmental heterogeneity in the marine gobies Pomatoschistus minutus and P. lozanoi (Gobiidae, Teleostei)

Genetic variation was assayed at 14 allozyme loci in estuarine, coastal and offshore samples of lozano's goby, Pomatoschistus lozanoi and the sand goby, P. minutus. Samples were taken from locations on the Belgian Continental Shelf and in the Schelde estuary with a range of environmental heterogeneity. We evaluate whether any differences in (1) the degree of genetic variation and (2) allele frequencies at the various loci exist within samples occurring in various habitats on the BCS and in the Schelde estuary. No significant differences in levels of genetic diversity were recorded between estuarine, coastal and offshore samples in either species. A temporally stable clinal gradient in allele frequencies at the two-allele locus GPI-A(*) was observed in P. lozanoi, differentiating the samples in an estuarine, coastal and offshore group. We suggest that these differences might be maintained by balancing selection at locus GPI-A(*).     

Host manipulation of a freshwater crustacean (Gammarus roeseli) by an acanthocephalan parasite (Polymorphus minutus) in a biological invasion context

Several gammarid species serve as intermediate hosts for the acanthocephalan parasite Polymorphus minutus. This parasite influences gammarid behaviour in order to favour transmission to its ultimate host, generally a bird. We investigated this host manipulation in Gammarus roeseli, a gammarid species introduced in France 150 years ago which now coexists with several exotic species from different origins. In the field, vertical distribution of G. roeseli revealed a higher proportion of infected individuals close to the water's surface and the size distribution of infected gammarids revealed predation pressure on infected individuals. However, under laboratory conditions both infected and non-infected individuals remained benthic. The addition of a second gammarid, Dikerogammarus villosus, to the experimental device involved a vertical displacement of infected G. roeseli towards the water's surface. Dikerogammarus villosus, originating from the Ponto-Caspian basin, can be considered as an aggressive predator. The substitution of D. villosus with Atyaephyra desmarestii, a planktivore decapod, did not alter the gammarids' distribution, with both infected and uninfected G. roeseli staying benthic. Thus, biotic interactions between D. villosus and G. roeseli represent selective pressure encouraging the expression of manipulated behaviour in infected amphipods. Through manipulation, P. minutus was found to increase the survival of infected G. roeseli when faced with non-host predators and to make it more vulnerable to predation by the parasite's definitive host.     

A New Myxosporean Parasite, Kudoa Camarguensis N. Sp., Recorded On Two Goby Species (Teleostei: Pisces) In the Rhône Delta (Mediterranean Sea, France)

ABSTRACT. A new species of myxosporean of the genus Kudoa was found in the muscle tissues of Pamatoschistus minutus (Pallas, 1770), a migratory fish species from the Mediterranean Sea. Previously this Kudoa species had been observed in Pomatoschistus microps (Krøyer, 1838), a sedentary species of the Vaccarés lagoon system. the parasite caused rapid myoliquefaction of the affected muscles within one hour after host death. Using both light and electron microscopy, whitish, longitudinal, spindle-shaped bodies, 2-5 mm long and 1-2 mm wide, were found in the caudal peduncle or in muscle tissues of the abdominal wall. Spore length, width, and thickness (n = 30) ranged between 4.4-6.0 üm, 6.4-8.0 üm, and 4.4-6.0 üm, respectively, while the polar capsules averaged 2.5 times 1.5 üm (n = 30). Anatomic comparison with other myxosporean parasites enabled us to determine this to be a new species that we name Kudoa camarguensis n. sp.     

Carotenoid-based colour of acanthocephalan cystacanths plays no role in host manipulation

Manipulation by parasites is a catchy concept that has been applied to a large range of phenotypic alterations brought about by parasites in their hosts. It has, for instance, been suggested that the carotenoid-based colour of acanthocephalan cystacanths is adaptive through increasing the conspicuousness of infected intermediate hosts and, hence, their vulnerability to appropriate final hosts such as fish predators. We revisited the evidence in favour of adaptive coloration of acanthocephalan parasites in relation to increased trophic transmission using the crustacean amphipod Gammarus pulex and two species of acanthocephalans, Pomphorhynchus laevis and Polymorphus minutus. Both species show carotenoid-based colorations, but rely, respectively, on freshwater fish and aquatic bird species as final hosts. In addition, the two parasites differ in the type of behavioural alteration brought to their common intermediate host. Pomphorhynchus laevis reverses negative phototaxis in G. pulex, whereas P. minutus reverses positive geotaxis. In aquaria, trout showed selective predation for P. laevis-infected gammarids, whereas P. minutus-infected ones did not differ from uninfected controls in their vulnerability to predation. We tested for an effect of parasite coloration on increased trophic transmission by painting a yellow-orange spot on the cuticle of uninfected gammarids and by masking the yellow-orange spot of infected individuals with inconspicuous brown paint. To enhance realism, match of colour between painted mimics and true parasite was carefully checked using a spectrometer. We found no evidence for a role of parasite coloration in the increased vulnerability of gammarids to predation by trout. Painted mimics did not differ from control uninfected gammarids in their vulnerability to predation by trout. In addition, covering the place through which the parasite was visible did not reduce the vulnerability of infected gammarids to predation by trout. We discuss alternative evolutionary explanations for the origin and maintenance of carotenoid-based colorations in acanthocephalan parasites.     

Length–weight relationships of 17 less-studied fish species from Çanakkale, Marmara region of Turkey

This study reports length–weight relationships for 17 cryptic and rare species, Aidablennius sphynx , Arnoglossus kessleri , Callionymus pusillus , Callionymus risso , Clinitrachus argentatus , Echiichthys vipera , Gobius geniporus , Gobius paganellus , Labrus viridis , Lepadogaster lepadogaster , Ophidion barbatum , Pomatoschistus minutus , Pomatoschistus pictus , Salaria pavo , Tripterygion delaisi , Tripterygion tripteronotus and Zebrus zebrus from Çanakkale near shores in the Marmara region of Turkey.     

Vacuolated prolactin cells in the pituitary gland of several marine teleosts

A morphometric study of prolactin cell ultrastructure in the pituitary gland of the Corkwing wrasse, Crenilabrus melops L., showed that cytoplasmic vacuoles, which accounted for 25% of the cell volume, were associated with signs of decreased secretory activity. The Golgi apparatus, mitochondria and rough endoplasmic reticulum, all contributed relatively little to the total cell volume, and there was no sign of secretory-granule release by exocytosis. All vacuoles were intracellular and membrane-bound, and probably derived from rough endoplasmic reticulum, Golgi apparatus and the nuclear envelope. It is thought that smaller vacuoles coalesce to form larger ones. The secretory granules were small and sparse, and this could account for the chromophobia of prolactin cells in light-microscopy preparations. Similar vacuoles were reported in the prolactin cells of the gobiid fish, Chaparrudo flavescens, Pomatoschistus pictus, Pomatoschistus minutus and Pomatoschistus microps . The vacuoles in Chuparrudo were of similar ultrastructure to those in Crenilabrus .