Interaction between exopolysaccharide and oxygenation levels on habitat selection in the sole Solea solea (L.)

A habitat selection experiment was conducted to examine the behavioural response of sole Solea solea to a combination of sediment quality (exopolysaccharides, EPS-free, i.e. 0 mg l⁻¹ and EPS-rich, i.e. 4 mg l⁻¹) and water oxygenation level (100 and 35% air saturation). The distribution of sole was influenced differently by the type of substratum depending on the water oxygenation level. In normoxia, sole settled preferentially on sand whereas under hypoxic conditions, sole settled preferentially on the muddy substratum. In order to explain these apparently counter-intuitive observations, it is proposed that, via cutaneous respiration, young sole are able to take advantage of the large quantities of oxygen produced by microphytobenthic organisms present in the upper few millimetres of muddy substratum.     

Chemical nature of feeding stimulants for the juvenile Dover sole, Solea solea (L.)

Juvenile Dover sole, Solea solea , were weaned from a live food, Artemia salina nauplii, on to a casein-based particulate diet flavoured with flesh of the mussel, Mytilus edulis . These fish would not accept an unflavoured casein-based diet but readily ate the diet flavoured with either mussel flesh or a mixture of pure chemicals whose composition was based on an analysis of the low-molecular weight fraction of mussel flesh. The rate of growth and survival over a seventy-seven day period was essentially the same on either of the flavoured diets. The active constituent in the mixture of pure chemicals was identified as glycine betaine for fish of wet weight exceeding 50g while glycine betaine with certain L-amino-acids was required for fish of about 2.5 g wet weight. These results are discussed in relation to the known feeding behaviour and food preferences of the wild Dover sole.     

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.     

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.     

Bio‐energetics underpins the spatial response of North Sea plaice (Pleuronectes platessa L.) and sole (Solea solea L.) to climate change

Climate change is currently one of the main driving forces behind changes in species distributions, and understanding the mechanisms that underpin macroecological patterns is necessary for a more predictive science. Warming sea water temperatures are expected to drive changes in ectothermic marine species ranges due to their thermal tolerance levels. Here, we develop a mechanistic tool to predict size‐ and season‐specific distributions based on the physiology of the species and the temperature and food conditions in the sea. The effects of climate conditions on physiological‐based habitat utilization was then examined for different size‐classes of two commercially important fish species in the North Sea, plaice, Pleuronectes platessa, and sole, Solea solea. The two species provide an attractive comparison as they differ in their physiology (e.g. preferred temperature range). Combining dynamic energy budget (DEB) models with the temperature and food conditions estimated by an ecosystem model (ERSEM), allowed spatial differences in potential growth (as a proxy for habitat quality) to be estimated for 2 years with contrasting temperature and food conditions. The resulting habitat quality maps were in broad agreement with observed ontogenetic and seasonal changes in distribution as well as with the recent changes in distribution which could be attributed to an increase in coastal temperatures. Our physiological‐based model provides a powerful tool to explore the effect of climate change on the spatio‐temporal fish dynamics, predict effects of local or broad‐scale environmental changes and provide a physiological basis for observed changes in species distributions.     

The influence of prey size,sediment thickness and fish size on consumption in common sole (Solea solea L.)

This study determined prey consumption in common sole as a function of prey size (0–0.5, 1–1.5, 2–2.5 and 4–5 g), sediment thickness (20 cm and 2 cm) and fish size (50 g, 125 g or 300 g). Prey consumption (in numbers of prey eaten per fish per day) was reduced with increasing prey size and sediment thickness, and was increased with increasing fish size (p < .001 for all factors). All 3 factors showed significant two way interactions (p < .001) when expressed in numbers of prey eaten. Prey consumption decreased with prey size when prey could not escape by burying (2 cm of sediment thickness) irrespective of fish size. We suggest that increasing effort to ingest and handle larger prey played a role. Prey consumption increased with fish size when prey could not bury (2 cm of sediment thickness). However, when prey was able to bury (at 20 cm sediment thickness) prey consumption was similar irrespective of fish size (p < .001 for interaction fish size × sediment). This interaction suggests that with increasing fish size there is an increasing mismatch between foraging adaptation and prey burial depth. This may explain the dominance of crustaceans in the diet of adult common sole in nature, despite the high abundance of polychaetes.     

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.     

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.     

The intestinal microflora of farmed Dover sole (Solea solea) at different stages of fish development

The bacterial flora in the intestines of farmed Dover sole ( Solea solea L.) was investigated at different stages of fish development in relation to different diets fed under farming conditions. The flora of water and diets were also analyzed. The dominant generic group isolated from all water samples examined was Pseudomonas Gp IV/ Alcaligenes. The same generic group was dominant in homogenized pelagic larvae indicating a relationship between the microflora of those young fish and that of tank water. In older fish a steady increase in the percentage incidence of Vibrio / anaerogenic Aeromonas was observed in the intestines as fish grew and diets changed. The Vibrio /anaerogenic Aeromonas group was dominant in Lumbricillus rivalis used as an intermediate diet and the increased incidence of this group in fish intestines occurred after fish were fed this diet. The composite pellet used for ongrowing carried a high percentage of Gram positive micro-organisms and there was no relationship between the pellet microflora and that found in the fish intestines.     

Observations on the ultrastructure of Haemogregarina simondi in the marine flatfish Solea solea (L.).

Developing stages of Haemogregarina simondi from the marine fish Solea solea (L.) were examined by electron microscopy. Merozoites lay in a parasitophorous vacuole and were bound by a pellicle of three unit membranes beneath which lay a ring of 45--61 microtubules. The cytoplasm contained 4--6 rhoptries, more than 169 micronemes, several mitochondria, and amylopectin granules. A conoid and one polar ring were observed at the anterior end. Intraleucocytic and intraerythrocytic schizonts with up to eight merozoites were described also. Intraerythrocytic and free gametocytes were characterized by distinct refractile bodies and a pellicle consisting of only two unit membranes. The number of micronemes was in excess of 194. The results were discussed in comparison with other members of the Haemogregarinidae.     

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.     

Metallic and metalloid elements in various developmental stages of Amanita muscaria (L.) Lam

There is growing evidence that mushrooms (fruiting bodies) can be suitable for biogeochemical prospecting for minerals and as indicators of heavy metal and radioactive contaminants in the terrestrial environment. Apart from the nutritional aspect, knowledge of accumulation dynamics and distribution of elements in fruiting bodies, from emergence to senescence, is essential as is standardization when choosing mushroom species as potential bioindicators and for monitoring purposes. We studied the effect of fruitbody developmental stage on the contents of the elements (Li, K, V, Cr, Mn, Mg, Co, Ni, Cu, Zn, As, Rb, Sr, Ag, Al, Cd, Sb, Cs, Ba, Pb, Tl and U) in the individual parts of the Amanita muscaria fruiting body. Elements such as K, Mg, Mn, Ni, Co, Cu, Zn and Se remained similar throughout all developmental stages studied, however for K, differences occurred in the values of caps and stipes, as expressed by the cap to stipe concentration quotient (index Q_C/S). The other elements quantified, i.e., Li, V, Cr, As, Rb, Sr, Ag, Al, Cd, Sb, Cs, Ba, Pb, Tl and U are considered as nonessential or toxic (with the exception of V in A. muscaria). Their accumulation in the fruiting bodies and their distribution between cap and stipe did not show a uniform pattern. Pb, Sb, Tl, Ba, Sr, Li, Rb and Cs decreased with increasing maturity of the fruitbodies, implying that translocation, distribution and accumulation in stipes and caps was not a continuous process, while V, Cr, As, Ag, Cd, and U remained at the same concentration, similarly to the essential elements. Our results for A. muscaria confirm that elemental distribution in different parts of fruiting bodies is variable for each element and may change during maturation. Soil properties, species specificity and the pattern of fruitbody development may all contribute to the various types of elemental distribution and suggest that the results for one species in one location may have only limited potential for generalization.     

Sugar metabolism and developmental stages of rubber tree (Hevea brasiliensis L.) seeds

Changes in the concentration of sugars and sucrose metabolism enzymes can characterize the developmental stages of a seed. In recalcitrant species such as Hevea brasiliensis L., little is known about these changes. We aimed to evaluate the three main stages of development of rubber tree seeds – histodifferentiation, cell elongation and accumulation of reserves. The activities of acid and neutral invertases (E.C. 3.2.1.26) and sucrose synthase (EC 2.4.1.13), and the concentrations of reducing sugars (RS), total soluble sugars (TSS) and sucrose (Suc) were determined concomitantly with the histochemical and anatomical evaluation of seed structure. Histodifferentiation in rubber tree seeds occurs up to 75 days after anthesis (DAA). The concentration of RS is high and of Suc is low during seed histodifferentiation, which occurs along with a visible increase in the number of cell divisions. After that period, there is an increase in the concentration of Suc (mg g^?1) and in the number and size of starch granules, and a decrease in the concentration of RS (mg g^?1). At that point, cell elongation occurs. At 135 DAA, there is an inversion in the concentration of these two sugars and an increase in reserve accumulation. Thus, in seeds of the evaluated clone, the period up to 75 DAA is characterized as the histodifferentiation stage, while from that time up to 120 DAA the cell elongation stage takes place. The final stage of seed maturation and reserve accumulation begins at 135 DAA, and the seed, including the embryo, is completely formed at 175 DAA.     

Comparative xenobiotic metabolism capacities and pesticide sensitivity in adults of Solea solea and Solea senegalensis

The measurement of enzymatic activities involved in xenobiotic biotransformation was carried out in adults of Solea solea and Solea senegalensis. The hepatic enzymes analysed were cytochrome P450 (CYP) related activities using eight fluorometric substrates and carboxylesterases (CbE). The conjugating activities of glutathione S-transferase (GST) and UPD-glucuronosyltransferase (UDPGT) were also assessed. Specific mammalian inhibitors were used as diagnostic tools for related activities of CYP1A (α-naphthoflavone; αNF), CYP2B6 and CYP2C19 (ticlopidine) and CYP3A4 (ketoconazole). The in vitro sensitivity to organophosphorous pesticides (OP) was tested in the S10 homogenate of brain (acetylcholinesterase-AChE) and liver (CbE). Furthermore, the pesticide chlorpyrifos oxon (CLPO) was used to explore the OP sensitivity of CbE of both species in two subcellular fractions (microsomes and cytosol), using two substrates. Overall, only two parameters confirmed species differences: EROD and cytosolic CbE being significantly elevated (p < 0.05) in the common sole, S. solea. A high inhibition of CYP1A related activities using several fluorometric substrates (ER, MR and CEC) after in vitro incubation with αNF confirmed all measure CYP1A1-related activities whereas ketoconazole was more specific for BFCOD (CYP3A4). Pesticide sensitivity was similar for brain AChE but hepatic CbE had a protective role that was species and pesticide dependent.     

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.     

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.     

Parasitic copepods of the common sole, Solea solea (L.), from the Eastern Mediterranean coast of Turkey

There is increasing interest in the common sole, Solea solea (Linnaeus), as an alternative fish species in aquaculture in the Mediterranean region, and parasitic copepods are a potential hazard for farmed finfish. This paper provides taxonomic information on two species of sea lice (family Caligidae) collected from S. solea in eastern Mediterranean waters off the Turkish coast. Caligus brevicaudatus A. Scott, 1901 and Caligus apodus (Brian, 1924) were both found and this is the first report of C. brevicaudatus in Turkish waters. The discovery of C. apodus on S. solea is a new host record. Key diagnostic characters of both species are reported, supported by light and scanning electron microscopy observations. During a 12-month survey a prevalence of 28% was recorded for C. brevicaudatus, whereas for C. apodus peak prevalence was much lower (3%).     

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.