Identification of a host-associated species complex using molecular and morphometric analyses,with the description of Gyrodactylus rugiensoides n. sp. (Gyrodactylidae,Monogenea)

Gyrodactylus rugiensis was originally described as a parasite occurring on the marine gobies Pomatoschistus minutus and Pomatoschistus microps. In our preliminary survey this species was also frequently found on Pomatoschistus pictus and Pomatoschistus lozanoi. Subsequent molecular analysis of the internal transcribed spacers rDNA region revealed that this parasite actually represents a complex of two apparently cryptic species, one restricted to P. microps and the other shared by P. minutus, P. lozanoi and P. pictus. Morphometric analyses were conducted on 17 features of the opisthaptoral hard parts of specimens collected from all four host species. Standard discriminant analysis showed a clear separation of both genotypes by significant differences in marginal hook and ventral bar features. Statistical classifiers (linear discriminant analysis and nearest neighbours) resulted in an estimated misclassification rate of 4.7 and 3.1%, respectively. Based on molecular, morphological and statistical analyses a new species, Gyrodactylus rugiensoides is described. This species seems to display a lower host-specificity than generally observed for Gyrodactylus species as it infects three sympatric host species.However, seasonal and host-dependent morphometric variation is shown for G. rugiensoides collected on P. pictus. Host-switching and gene flow might be important factors preventing speciation on closely related and sympatric host species. The presence of host associated species complexes in this Gyrodactylus-Pomatoschistus system is also confirmed by the presence of two host-dependent genotypes within G. micropsi found on P. minutus and P. lozanoi, and P. microps, respectively. By comparing host and parasite phylogeny, phylogenetic and ecological factors influencing host-specificity are discussed.     

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 .     

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.     

Sublittoral abundance and food consumption of Baltic gobies

Two small demersal fishes, the sand goby Pomatoschistus minutus and the common goby Pomatoschistus microps , were quantified on soft bottoms at 20–40 m depth in the Baltic Sea, using a camera placed above the bottom. The largest numbers of gobies were seen following the settlement of young in late summer and autumn. Most recorded fishes were sand gobies. An annual average of 4·7 individuals m⁻²(0·24 g dry mass m⁻²) was recorded in 1983–1985 and 2·5 individuals m⁻²(0·13 g m⁻²) in 1997–1998. Using these densities, the annual goby food consumption was estimated to 100 kJ m⁻² in 1983–1985 and 50 kJ m⁻² in 1997–1998, corresponding to most of the annual macrobenthos production available to the gobies. The resulting goby production, assumed equal to 25% of the food consumed, must have been an important food source for the larger fishes occasionally recorded in the photographs.     

Current and historic gene flow of the sand goby Pomatoschistus minutus on the European Continental Shelf and in the Mediterranean Sea

Phylogeographical patterns of the sand goby Pomatoschistus minutus (Gobiidae, Teleostei) were studied by means of sequence and single-stranded conformational polymorphism analysis of a 283-bp fragment of the cytochrome b locus of the mtDNA. A total of 228 individuals sampled at 13 sites throughout the species's distributional range revealed a moderate level of diversity and a low but significant level of overall genetic differentiation at all but one site. The goby sample from the Adriatic Sea differed in sequence by approximately 10% from the Atlantic P. minutus and is thought to belong to a cryptic species of the genus Pomatoschistus . Limited genetic differentiation with a weak pattern of isolation-by-distance was recorded throughout the distributional range of the typical P. minutus . Phylogeographical analysis suggested a contiguous range expansion in the Atlantic and Baltic basins during the Eemian and evidence for a glacial refugium in the southern North Sea during the Weichselian. In P. minutus from the western Mediterranean Sea a high number of endemic haplotypes as well as the most common Atlantic haplotype were recorded in appreciable frequencies. This might be explained by secondary contact between different mitochondrial lineages, which evolved in allopatry.   © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 83 , 561–576.     

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.     

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.     

Paleoclimatic history and vicariant speciation in the "sand goby" group (Gobiidae, Teleostei)

Vicariant and climatic cycling speciation hypotheses of the 'sand gobies' belonging to the genera Pomatoschistus, Gobiusculus, Knipowitschia, and Economidichthys are tested using molecular phylogenies constructed of nuclear DNA (ITS1 locus) and mitochondrial DNA (12S and 16S fragments). These gobies are among the most abundant in the Eastern Atlantic-Mediterranean region, and play an important role in the ecosystem. Considerable ITS1 length differences, primarily due to the presence of several tandem repeats, were found between species and even within individuals. Therefore, phylogenetic analyses focused on fragments of the 12S and 16S mtDNA region that have been sequenced for 16 goby taxa. The 'sand gobies' clustered as a monophyletic group as proposed on morphological grounds. However, G. flavescens, E. pygmaeus, and K. punctatissima clustered within the Pomatoschistus species, pointing to a paraphyletic origin of these genera. Furthermore, the genetic divergence between P. minutus from the Adriatic Sea versus the Atlantic-Mediterranean region was as high as the divergence within the P. minutus complex, suggesting that P. minutus from the Adriatic Sea should be considered as a distinct species. The "star" phylogeny might suggest that these gobies evolved in a very short time period, possibly linked to the drastic alterations in the Mediterranean Sea during and immediately after the Messinian salinity crisis at the end of the Miocene. The freshwater life-style appeared monophyletic; equating its origin with the salinity crisis resulted in a molecular clock estimate of 1.4% divergence per million years. The last common ancestor probably occupied sandy bottoms and a coastal niche while several species subsequently adapted to new habitats (pelagic, freshwater or stenohaline). The origin of the shallowest clades dated back to the glacial cycling during the Pleistocene epoch.     

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.     

Big is better: the importance of size for reproductive success in male Pomatoschistus minutus (Pallas) (Pisces, Gobiidae)

The sand goby, Pomatoschistus minutus , was used in aquarium experiments to study the importance of body size for the ability of males to gain access to nest sites and mates. When several male and female P. minutus were allowed to reproduce together, on average, half of the males built nests, and half of those males obtained eggs in their nests. Males with nests were significantly larger than males without nests, and nests with eggs belonged to males larger than the males with empty nests. In another experiment, when two males were competing for one nest, the largest male occupied the nest when both fish were put into the aquarium simultaneously. However, when the smaller male had been allowed to establish a nest before the larger male was introduced, the small male could usually retain the nest. Males with a body length < 50 mm did not build nests at all in the early part of the breeding season. In female choice experiments, no preference for larger males was found. Thus, male-male competition for nest sites and behavioural differences between different sized males seem to be the main factors influencing the non-random mating success in male P. minutus.     

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.     

Life history traits of Pomatoschistus minutus in the Rhône Delta, France

Life history traits and migratory status of the Sand goby Pomatoschistus minutus were investigated in a brackish lagoon of the Rhône river Delta (Mediterranean Sea, south of France).     

A new West Balkanian sand-goby (Teleostei: Gobiidae)

A new species of West Balkanian freshwater sand-goby Pomatoschistus montenegrensis sp. nov. (Teleostei: Gobiidae) is described from the Morača River, southern Montenegro, and shown to be related to the euryhaline Adriatic Pomatoschistus canestrinii . The generic status of these two species is discussed with reference to the limits of Pomatoschistus (type Gobius minutus ) and the status of a subgenus [ Ninnigobius (type-species Gobius canestrini )]. The habitat and the local distribution of the new species are described.     

Effect of digestion on the δ13C and δ15N of fish-gut contents

Gut contents of sand goby Pomatoschistus minutus showed higher C and N isotope values than the food before consumption. This enrichment was more pronounced in the hindgut than in the foregut, probably because of preferential assimilation of ¹²C and ¹⁴N along the gastro-intestinal tract. The results indicated that the shift towards higher values in the alimentary canal occurs in the first 2 h after feeding.     

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.     

Sexually selected nest-building--Pomatoschistus minutus males build smaller nest-openings in the presence of sneaker males

Both natural selection and sexual selection may act on nest-building. We tested experimentally how different regimes of egg-predation and male-male competition influence nest-building before mating, using the marine fish sand goby, Pomatoschistus minutus. Males with sneaker males present built the smallest nest-openings, smaller than males held alone or with Pomatoschistus microps males (which may predate eggs and compete over nest-sites but not compete over fertilizations). Males with visual access to other nest-building males tended also to build smaller openings than males held alone or with P. microps. Males with egg-predators present built nests with openings not differing significantly from any other treatment. Our results indicate that the small nest-openings found in the sneaker male treatment are sexually selected through protection against sneaking or by female choice. Across treatments, time span before a male started to build his nest also explained variation in nest-opening width; males starting late built larger nest-openings.     

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.     

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(*).     

Tolerance, behaviour and oxygen consumption in the sand goby, Pomatoschistus minutus (Pallas), exposed to hypoxia

Behavioural and metabolic reactions of Pomatoschistus minutus (Pallas) exposed to various degrees of hypoxia were studied. At 15°C and 20‰ mortality was 50% at 15.2% oxygen saturation. Avoidance and oxygen saturation showed a linear inverse relationship. At levels lower than 60% saturation increased activity occurred; avoidance was significant at 30% saturation. Active, routine and standard MO₂ correlated linearly with weight at 6 and 15°C (salinity = 19‰). During hypoxia at 15°C routine MO₂ rose significantly at 60–50% and 40–30% saturation expressed either as MO₂ during longer periods at night or MO₂ at shorter intervals during the day. Standard MO₂ was unaffected by hypoxia at 15°C. Haemoglobin concentration was significantly increased when P. minutus was acclimatized to 35% saturation.     

Costly courtship or dishonest display? Intensely displaying sand goby males have lower lipid content

Intensely displaying sand goby Pomatoschistus minutus males had lower lipid content, indicating that signalling increased energy expenditure, or that low condition males increased their signalling effort. Display intensity correlated positively with nest‐defence and tended to correlate positively with filial cannibalism.