Caligus solea n. sp. (Copepoda: Caligidae) parasitic on the common sole Solea solea (Linnaeus) from the north-eastern Mediterranean off the Turkish coast

A new species of caligid copepod, Caligus solea n. sp., is described from the common sole Solea solea (Linnaeus) caught off the north-eastern Mediterranean coast of Turkey. Both sexes of the parasite were collected from all over the upper body surface of its host. The new species belongs to the macarovi-group of species as established by Boxshall & Gurney (Bull Br Mus (Nat Hist) (Zool), 39:161–178, 1980), with which it shares the following four characters: (i) leg 4 with two-segmented exopod, distal segment carrying three apical spines but no lateral spine; (ii) distal exopodal segment of leg 1 with three plumose setae posteriorly plus four distal margin elements, spine 1 naked, spines 2 and 3 with accessory process and spine 4 about twice length of the others; (iii) females with one-segmented abdomen while males with two-segmented abdomen; (iv) male maxilliped with myxal process opposing the tip of the subchela. However, the new species differs from its congeners within the macarovi-group in the number of sensillae on each papilla on and around the postantennal process, and also in the absence of serrations along the distal margin of the maxilla. This is the twenty-eighth species of Caligus to be reported from the Mediterranean Sea.     

Movements of juvenile Dover sole, Solea solea (L.), in the Tamar Estuary, South-western England

Solea solea start to appear in the Tamar Estuary in April–May when they are 10 mm long. The majority of young sole move into the estuary in June–July. Most sole leave the estuary towards the end of their second year in October–November, when they have reached a total length of 150–200 mm. The young O-group fish remain in the main channel, whereas the I-group move with the tide onto the mud-flats.
Approximately 7000 fish were marked by freeze-branding or dye-injection. The recaptures showed that there was an up-estuary migration in May–June. Most of the recaptures were taken at the site of original capture and there was only a limited movement of fish between adjacent mud-flat areas. Average growth rate tends to be greater in sole inhabiting the upper reaches of the estuary.     

Spatial variability in digenean metacercariae infection of 0-group common sole Solea solea among nurseries along the French Atlantic coast

We sampled 0-group sole juveniles (N = 174) in September and October 2003 in 9 major nurseries located along the French Atlantic coast (English Channel and Bay of Biscay). 0-group sole were infected with 3 genera of digenean metacercariae, Timoniella spp. (Acanthostomidae), Prosorhynchus crucibulum and Prosorhynchus sp. A (Bucephalidae), Podocotyle sp. (Opecoelidae), and an unidentified species of Digenea. Parasite infection levels in the English Channel nurseries were lower, and the community composition was different from Bay of Biscay nurseries. We hypothesize that the difference between geographic areas was due to differences in first intermediate host communities and, to a lesser extent, because of lower temperatures in the English Channel compared to the Bay of Biscay. For the Bay of Biscay nurseries, mean total parasite abundance was strongly negatively correlated with mean annual river flow. This relationship may be the result of the more upstream location of 0-group sole nurseries in estuaries so that fish were further from local points of parasite transmission than those in embayed nurseries. Digenean metacercariae load may be influenced by 3 major local factors, i.e. abundance and proximity of the first intermediate hosts and cercariae dispersal capacities.     

An electron microscopic examination of the gastrointestinal epithelium in Dover sole, Solea solea (L.)

The gastrointestinal tracts of adult and juvenile Dover sole, Solea solea (L.), were examined using scanning (SEM) and transmission electron microscopy (TEM). SEM showed little differentiation of the internal morphology of the gastrointestinal tract in adult fish, with longitudinally arranged mucosal folds present in all gut regions. Mucosal folds had a similar arrangement to that in the goldfish. Goblet cells were identified in the mucosal epithelium in all regions of the gut while microscopic ducts/pores of possible pancreatic origin were observed in the foregut region. SEM of juvenile gut samples showed a similar arrangement of longitudinal mucosal folds to that found in the adult fish. There was no visible evidence of goblet cells or secretory ducts/pores in any region of the juvenile gastrointestinal tract.
In TEM it was observed that apical microvilli were a feature of epithelial cells from all regions of the gastrointestinal tract in adult and juvenile Dover sole. Cells from the distal regions of the adult and juvenile gut showed invaginations and vacuolation of the apical cytoplasm. A high degree of vacuolation in cells from the juvenile hindgut-rectum indicated the possible occurrence of intracellular digestion of absorbed nutrients in this gut region.     

The effect of hypoxia on ventilation frequency in startled common sole Solea solea

Ventilation frequency (F_V) in motionless common sole Solea solea was measured before and after a startling stimulus in normoxia and in hypoxia (15% air saturation). Startling reduced F_V in normoxia (from mean ±s.e. 41 ± 3·3 beats min^?1 to near zero, i.e. 2·0 ± 1·8 beats min^?1) and in hypoxia (from mean ±s.e. 80 ± 4·4 to 58·8 ± 12·9 beats min^?1). It is suggested that the maintenance of high F_V in hypoxia may increase the probability of detection by predators compared to normoxia.     

Coupling of heart rate and locomotor activity in sole, Solea solea (L.), and bass, Dicentrarchus labrax (L.), in their natural environment by using ultrasonic telemetry

Field measurements of distance moved and heart rate in sole, Solea solea (L.) and bass, Dicentrarchus labrax (L.) using ultrasonic telemetry revealed two different strategies. Heart rate in the sole increases during activity, accurately reflecting fluctuation in metabolic rate and so can be used as a measure of metabolic rate in the field. In contrast, the relatively stable value of the heart rate in bass during the whole tracking period whatever the activity level suggests that in this species heart rate in the field cannot be associated with metabolic rate determination.     

Malnutrition may affect common sole (Solea solea L.) growth, pigmentation and stress response: molecular, biochemical and histological implications

In the last decades there have been several evidences that traditionally used live preys like rotifers and Artemia salina have nutritional deficiencies that result in a general decrease of fish health, causing anomalies in the development, in growth and in pigmentation. In this study a partial of total replacement of traditional live preys with preserved copepods that represent the natural food of the larvae was evaluated during Solea solea culture. In this study a positive effect of co-feeding preserved copepods in sole larviculture was observed since larvae fed this diet growth and survived better, showed a better tolerance to captive conditions and had a better response to the final thermal/density stress-test with respect to larvae fed a traditional diet. Morphometric data were fully supported by molecular and biochemical ones. Moreover, liver histological investigations, revealed that the inclusion of preserved copepods in the larval diet was able to improve lipid assimilation. In conclusion, preserved copepods may be considered a suitable food for sole when used as a supplement to the traditional diet based on rotifers and Artemia nauplii.     

The mode of hatching of the monogenean Entobdella soleae, a skin parasite of the common sole (Solea solea).

Observations have been made on the hatching behaviour of the larva of the monogenean Entobdella soleae and on the effects of various enzymes on the opercular cement of the egg. These observations indicate that hatching is brought about by a proteolytic hatching fluid which is produced by two pairs of ventral head glands and spread over the inner surface of the opercular joint by rotation of the larva about its longitudinal axis. After hatching there is no trace of the ventral head glands. Similar glands are present before hatching in the oncomiracidium of E. hippoglossi.     

Influence of DNA isolation from historical otoliths on nuclear-mitochondrial marker amplification and age determination in an overexploited fish, the common sole (Solea solea L.)

Historical otolith collections are crucial in assessing the evolutionary consequences of natural and anthropogenic changes on the demography and connectivity of commercially important fish species. Hence, it is important to define optimal protocols for purifying DNA from such valuable information sources while avoiding any damage to the physical structure of the otolith. Before being able to conclude on the harmlessness of a method, it is important to validate protocols on different kinds of otoliths by testing purification methodologies under standardized conditions. Here we compare the effect of two DNA extraction methods on the success in identifying the age in an overexploited marine fish, the common sole (Solea solea L.). To ensure optimal future population genetic and demographic analyses, we assessed DNA quantity and tested the DNA quality by investigating the amplification success of a mitochondrial and nuclear marker. Our results show that the choice of the DNA extraction method had a significant effect on the success of using these otoliths in age and growth analyses. Standard commercial and published protocols resulted in a severe damaging of the otolith structure, hampering accurate preparation and analyses of the morphological structures of the otoliths. Shortening the lysis time and lowering the EDTA (ethylene diamine tetraacetic acid) and SDS (sodium dodecylsulphate) concentration turned out to be beneficial for the stability of otolith structure, while maintaining an overall high DNA quality measured through polymerase chain reaction amplification success. We therefore recommend that care should be taken when choosing the extraction method for a molecular study on archived samples, in order to enable the maximal use of information embedded in historical material.     

The influence of sand on the estimation of resting metabolic rate of juvenile sole, Solea solea (L.)

The influence of a sandy substrate on the estimation of the resting metabolic rate of juvenile sole was determined using a static type of respirometer. Oxygen consumption of fish deprived of sand was almost double that of fish provided with sand.     

Parasitic infection of sole Solea solea by Prosorhynchus spp. metacercariae (Digenea, Bucephalidae) in Atlantic nurseries under mussel cultivation influence

Cysts of metacercariae were obtained on 2 dates from juvenile sole Solea solea sea-sampled in an area of mussel cultivation (Pertuis Charentais, Bay of Biscay, France). An initial assessment of parasite genus and infestation level was based on 192 cysts extracted from 2 fish samples, taken in August (n = 20) and December 2000 (n = 14). Our results confirmed the sole as second intermediate host of bucephalid trematodes of the genus Prosorhynchus, which has not previously been noticed in Atlantic stocks. Prevalence, ca. 65% on both dates, indicated an substantial infestation of these small fish, with a mean abundance of parasites increasing from August (3.3 +/- 1.1) to December (8.1 +/- 3.4). Cysts were localised in all body parts of the host, and positioning varied depending on sampling date. However, the cephalic area was always the most infested (72.7 and 49.1% in August and December, respectively). Parasite measurements suggested a protracted infestation process, which may be initiated in spring during sole settlement. Most of the largest metacercariae had the rhynchus characteristics of P. crucibulum, though the possibility of them being other species (P. squamatus, P. aculeatus) could not be excluded. As Mytilus edulis is the first intermediate host of Prosorhynchus spp., possible relationships between mussel culture and sole parasitosis are discussed.     

Metabolism in marine flatfish. II. Protein digestion in Dover sole (Solea solea L.)

The digestive tracts of adult and juvenile Dover sole were examined for protease activities. A pepsin-like protease with an optimal pH value of 1.7 predominated in the stomach region, but the main endoprotease action in the foregut, midgut and hindgut regions was optimal in the range of pH 9.5-10.5 and showed good activity towards elastin orcein. Experiments using synthetic substrates suggested the presence of chymotrypsin- and trypsin-like activities optimal between pH 7 and 8. Collagenase activity was also shown to exist in this pH region. The presence of enzymes corresponding to carboxypeptidases a and b and leucine aminopeptidase was indicated. The possible significance of these results to the farming of Dover sole is discussed.     

Comparison between the life cycles of two Soleidae, the common sole, Solea solea, and the thickback sole, Microchirus variegatus, in the Bay of Biscay (France)

The life history of two Soleidae, the common sole, Solea solea, and the thickback sole, Microchirus variegatus, were compared in the Bay of Biscay in an attempt to set out factors which could explain settlement styles known to be different between juveniles of each species. Common sole juveniles had been shown to depend on coastal and estuarine nurseries, and the thickback sole to develop in open-sea nurseries, although the spawning grounds and spawning season of both species overlapped (offshore and at springtime, respectively). For this study, data on adult, juvenile and larva distributions were obtained from cruises carried out in the Bay of Biscay during the last decade. In addition, growth rate of larvae and planktonic interval duration, estimated by means of otolith increment analysis, were compared, as well as literature-derived information on behaviour of larvae. By comparing larval features, it appeared that thickback sole did not obtain the advantage of a slightly longer pelagic life span for an increased dispersal, due to an early shift to benthic behaviour. Other evidence was given by more specifically stated distributions that the further and deeper offshore spawning of M. variegatus, compared to S. solea, was a likely key-factor of the juvenile settlement process. This could explain why the thickback sole, contrary to the common sole, never reach coastal areas and thus settle offshore, in waters deeper than 30 m.     

Effect of temperature and salinity on the gastric evacuation of juvenile sole Solea solea and Solea senegalensis

The juveniles of Senegal sole, Solea senegalensis, Kaup 1858, and common sole, Solea solea (Linnaeus 1758) concentrate in estuarine and coastal nurseries of widely differing temperatures and salinities. Yet, little is known about the effect of these physiologically important variables on the gastric evacuation rates of these species. Gastric evacuation experiments were performed on juveniles of S. senegalensis and S. solea. Three temperatures were tested, 26, 20 and 14°C at a salinity of 35‰. A low salinity experiment was also carried out at 15‰, at 26°C. Experimental conditions intended to reflect conditions in estuarine and coastal nurseries where juveniles of these species spend their first years of life. The relation between stomach contents and time was best described by exponential regression models for both species. An analysis of covariance (ancova ) was performed in order to test differences in evacuation rate due to temperature and salinity (slope of evacuation time against stomach contents) for each species. While increasing temperature increased evacuation rates in both species (although not at 26°C in S. solea), the effect of low salinity differed among species, leading to a decrease in gastric evacuation rate in that of S. senegalensis and an increase in S. solea. Differences in gastric evacuation rate between species were related to its metabolic optimums and to its distribution in the nursery area where fish were captured. Implications for the habitat use of estuarine and coastal nurseries are discussed.     

Metabolism in marine flatfish--III. Measurement of elastase activity in the digestive tract of Dover sole (Solea solea L.)

A critical assessment of different methods for measuring elastase activity in crude preparations has been made using whole intestinal homogenates of Dover sole. The use of the natural substrate elastin or its dyed derivatives gave optimal pH values in the alkaline region (pH 9.4-9.8) whereas artificial substrates showed optimal hydrolysis nearer neutrality in the region pH 8.1-8.2. Exoproteases may interfere with certain assay procedures. The properties of Dover sole elastase have been further investigated using chromatographic techniques which indicated that the main elastase activity has a molecular weight of approximately 19,500 and an isoelectric point in the region of pH 5.7.