Characterization of eight microsatellite markers in the white sea bream, Diplodus sargus (Teleostei, Sparidae)

The white sea bream, Diplodus sargus (Teleostei, Sparidae), is a species with a high commercial importance in Mediterranean aquaculture. There is currently little information available about the genetic characteristics of cultured populations. In this survey, we have developed eight polymorphic microsatellites for the white sea bream using an enriched genome library protocol. All of them were polymorphic in the 67 individuals tested, 32 of which were wild specimens, and 35 were individuals from a captive F(1) broodstock. These markers can potentially be useful tools for use in population genetic studies.     

Isolation and characterization of nine polymorphic microsatellite markers in the two‐banded sea bream (Diplodus vulgaris) and cross‐species amplification in the white sea bream (Diplodus sargus) and the saddled bream (Oblada melanura)

We have developed nine new microsatellite markers for the two‐banded sea bream (Diplodus vulgaris) from an enriched genome library protocol. All these loci are polymorphic, with mean allelic diversity of 13 (range 5 –21), and expected and observed heterozygosities from 0.641 to 0.932 and 0.428 to 0.914, respectively. Cross‐species tests in two close‐related species of the genus Diplodus (D. sargus and O. melanura) revealed successful amplifications at 8 out of 9 loci, with mean allele number of 4.75 (range 2–8) and 5.50 (range 3 –10), respectively. These results are consistent with the close phylogenetic relationships between the three species, indicating this set of primers might prove useful for studying the levels of genetic diversity and population differentiation in these three species and in other phylogenetically close species of the genus Diplodus and Sparus.     

Further studies on the sexuality of the hermaphroditic teleost Diplodus sargus, with particular reference to protandrous sex inversion

Histological events associated with the process of sex inversion are described in the protandrous teleost Diplodus sargus reared in captivity. This species possesses typical sparid ovotestes in which the testis and ovary occur in separate zones. After spawning, the testicular tissue within the bisexual gonad undergoes regressive changes such as spermatogonial degeneration, deposition of yellowish pigment and proliferation of connective cells and fibres. At the same time ovarian tissue begins to develop. Transitional individuals were 26–7% of examined fish, while the other 73–3% were functional males.     

Parasites in marine protected areas: success and specificity of monogeneans

The effect of a marine protected area on the Lamellodiscus spp. monogenean community was tested by comparing the communities of parasites of Diplodus sargus inside and outside of the area. A total of 104 D. sargus were dissected harbouring 1280 monogeneans from the genus Lamellodiscus and belonging to 11 species. No modification in the global parasite community linked with the protection of the host populations was revealed. The most abundant and less specific parasite species, Lamellodiscus elegans , however, increased its abundance in the protected area. A significant relationship was found between parasite host range and the percentage of infected hosts. A significant relationship also occurred between epidemiological and genetic distances for the parasite species found. The results are discussed in term of parasite success and specificity and the importance of taking into account parasitism in the biological conservation of hosts.     

Genetic differentiation of white sea bream within the Lion's Gulf and the Ligurian Sea (Mediterranean Sea)

An allozymic study was conducted on 190 individuals of white sea bream, Diplodus sargus sampled on five locations throughout the Lion's Gulf and the Ligurian Sea (Mediterranean Sea), in order to identify genetic structure. Electrophoretic surveys carried out on muscle and liver tissues identified 25 loci of which 12 were polymorphic ( P 0.95). G -test analysis shows significant differences on allelic frequencies between the five stations at six loci. Examination of the spatial structure was performed using Nei's distances and F statistics, and indicated significant genetic differences between three groups. A group which clustered Blanes (Spain), Marseille (France) and Livorno (Italy) where the absence of genetic difference can be explained by migration of larvae and adults along a coastal 'continuum'. The Elba sample (Italy) shows genetic divergence from other samples and this difference may result from isolation due to limited migration of larvae and adults. Banyuls (France), differs from all other stations. Several hypotheses are proposed to explain genetic patterns including local current systems, larval dispersal, geographic isolation and historical effects, and variation in the size classes of sampled individuals between sites.     

Aerial and aquatic feeding in the silver arawana,Osteoglossum bicirrhosum

Synopsis The silver arawana, Osteoglossum bicirrhosum, hunts along shorelines and within flooded forests in the Amazon River basin and supplements its limited consumption of aquatic vertebrates by leaping from the water to obtain terrestrial and arboreal prey. We offered O. bicirrhosum prey both suspended above and submerged below the surface of the water. From high-speed digital recordings, we measured kinematic variables associated with the jaws, cranium, pectoral fins, and body during orientation and prey capture. Aquatic and aerial feeding events were kinematically distinct, with aerial events generally involving faster, larger movements and a distinct delay in the onset of lower jaw depression until the head had left the water. The comparatively large gape during leaping may facilitate prey capture by overcoming variability in the apparent position of the prey due to refraction, while the delayed onset of mouth opening may serve to reduce the effects of drag. This distinctive leaping behaviour allows exploitation of the terrestrial prey base, especially during seasonal inundation of the Amazon River basin when the aquatic food base is widely dispersed.     

A quantitative and histochemical study on melano-macrophage centres in the spleen of the teleost fish Diplodus annularis L.

The incidence and histochemical features of melano-macrophage centres in the spleen of the teleost fish D. annularis were studied with respect to starved and fed fish. Tissue catabolism following complete starvation appears to induce formation of new centres containing melanin and lipofuscin pigments. However, a change in the amount of melano-macrophages was also recorded in fed fish, suggesting that some other factor(s), internal to fish, may affect the presence of such structures within the spleen of D. annularis .     

The structure of the buccal cavity and pharynx in relation to the method of feeding of Reighardia sternae Diesing 1864 (Pentastomida)

The buccal complex of the haematophagous pentastomid Reighardia sternae has been described. It consists of a buccal capsule containing a prominent oral papilla communicating with a heavily chitinized pharynx which is crescentic in cross section. The chitinous skeleton of the buccal apparatus provides a framework for the attachment of various muscle systems, the contractions of which have been related to the mode of operation of the oral papilla and pharnyx during the feeding process. Particular emphasis was placed on the possible function of the oral papilla in the initial rupture of a blood capillary, and on the pumping action of the pharynx during the ingestion of the liberated blood.     

Kinematics of the pharyngeal jaws during feeding in Oreochromis niloticus (Pisces,Perciformes)

Cichlids possess a complex pharyngeal jaw apparatus, the osteological components of which are two upper pharyngeal jaws, articulating with the neurocranial base, and a single lower pharyngeal jaw. Quantitative cinera-diography revealed that pharyngeal food processing in Oreochromis niloticus involves transport, mastication, and swallowing, effected by cyclical pharyngeal jaw movements. Transport and swallowing occur by simultaneous retractions of both upper pharyngeal jaws. Food reduction (mastication) is effected by lower jaw elevation (compression) and protraction (shear) during upper jaw retraction. Each movement cycle contains a transport, reduction, and swallowing component, although their relative importance may vary within a feeding sequence. The upper and lower pharyngeal jaws show opposite anteroposterior movements during most of the cycle. Variations in the amplitudes and the durations of the different movement components reflect the consistency and the size of the food.     

Implication of the feeding limb morphology for herbivorous feeding in some freshwater cyclopoid copepods

1. The mouthparts of five species of adult planktonic cyclopoid copepods (Cyclops vicinus, C. abyssorum, Acanthocyclops robustus, Mesocyclops leuckarti and Thermocyclopscrassus), in particular the distance between setae and setules of the maxilliped (which can indicate the ability to retain small particles), were compared using electron and light microscopy. 2. The mesh‐sizes of the food‐collection grid formed by these setae and setules ranged between 4.6 and 13.2 μm; the area covered by the grid ranged between 6000 and 32 000 μm². 3. Mesh‐size was not simply correlated with body size. Cyclops abyssorum and M. leuckarti have the coarsest meshes and T. crassus the finest, while C. vicinus and A. robustus were intermediate. 4. The results suggest that cyclopoid copepods are able to retain particles in the size range of nanoplankton and that differences in mesh‐sizes between species may explain differences in the ability to subsist and reproduce on a diet of small algae.     

Organogenesis of the digestive tract in the white seabream,Diplodus sargus. Histological and histochemical approaches

The ontogeny of the digestive tract of the white seabream, Diplodus sargus during the larval development up to day 45 post-hatching (dph) has been studied using histological and histochemical techniques. The oesophageal goblet cells appeared around 6 dph and contained neutral and acid mucosubstances (PAS/diastase-PAS and Alcian Blue pH 2.5 positive reactions). An incipient stomach can be distinguished from 2 dph but the first sign of gastric gland development was detected around 13-15 dph, increasing in number and size by 22-23 dph. Gastric glands were concentrated in the cardiac stomach region and they had a high content of protein rich in tyrosine, arginine and tryptophan. Acidophilic supranuclear inclusions related to pynocitosis of proteins, were already observed in the intestinal cells of the posterior intestine around 4-6 dph (exogenous feeding) and they were present until 25 dph. The intestinal mucous cells appeared between 15-18 dph and contained a mixture of neutral and acid mucosubstances/glycoconjugates, carboxylated ones being more abundant than the sulphated ones. The stomach and gastric glands were fully developed by the first month of life marking the beginning of digestive features characteristic of the juvenile stage. Around 4-6 dph, glycogen, proteins and neutral lipids were observed in the granular cytoplasm of hepatocytes. Strongly acidophilic zymogen granules were also present, at this time, in the basophilic cytoplasm of the exocrine pancreatic acinar cells and contained abundant proteins, especially rich in arginine, tyrosine and tryptophan.     

Gut morphology,feeding rate and gut clearance in five species of caddis larvae

Feeding rate, the rate of movement of food through the gut and gut morphology of large larvae of five caddis species (Halesus radiatus, Hydropsyche instabilis, Polycentropus kingi, Rhyacophila dorsalis and Potamophylax cingulatus) were investigated in the laboratory. Following 72 hr starvation, P. cingulatus and H. radiatus larvae became satiated (refused prey offered directly to the mouthparts) after consuming 8–11 and 9–13 mayfly nymphs (Baetis rhodani, 3.5–4.6 mm) respectively. Hunger level affected prey consumption. In P. cingulatus, the number of prey consumed over 24 h (at 9.5–12 °C) increased with starvation periods from 0–72 h, but declined following longer starvation periods. Six clearly recognisable gut states (defined by the position of food material in various parts of the gut) can be identified at different times since commencement of a meal. As environmental temperature increased (from 8–12 °C to 15–17 °C), the rate of change of the gut state increased and the food retention time decreased in all species. Feeding periodicity (i.e. nocturnal/diurnal activity) in the field was estimated based on the evacuation rate and the gut state and environmental temperature at the known time of collection. Initiation of consumption of prey appeared to coincide with emptying of the foregut and proximal midgut, whereas actual feeding continued until complete satiation when some threshold fullness of the foregut had been reached. The length of time food was held in the foregut was positively correlated with increasing specialisation of the foregut (particularly elaboration of the proventriculus).     

Histopathology and cellular response in Enteromyxum leei (Myxozoa) infections of Diplodus puntazzo (Teleostei)

Enteromyxum leei is an intestinal parasite responsible for serious outbreaks in Mediterranean sharpsnout sea bream Diplodus puntazzo. E. leei infection was experimentally transmitted to healthy D. puntazzo (R) by cohabitation with infected donor fish. Haematological changes and histopathological damage were evaluated in relation to the course of infection. The prevalence of infection in R fish was 100% from day 10 post-exposure (p.e.) onwards, and the infection intensity and histopathological damage increased progressively. Different developmental stages were found in the infected intestines, including proliferative (stages 1–3) and sporogonic (stages 4 and 5) stages. Intestinal damage consisted of vacuolation, necrosis, detachment and sloughing of epithelium, and was correlated with the progression of the infection and with the development of the parasite. Sporogonic stages appeared from day 20 p.e. onwards. Initially, D. puntazzo seems to counteract the infection through the increase in leucocyte numbers, respiratory burst activity, haematopoietic activity and MMC. Two types of eosinophilic granular cells (EGC1 and EGC2) were detected in the intestinal epithelium and lamina propria. EGC1 numbers decreased with the progression of infection, whereas an increase in EGC2 occurred, mainly in the lamina propria. The involvement of the cellular immunity in the response of D. puntazzo to E. leei was demonstrated. The depletion of this response at a certain point of the infection could contribute to the high virulence of this myxozoan in this fish species.     

Immunohistochemical detection of ACTH and MSH cells in the hypophysis of the hermaphroditic teleost, Diplodus sargus

Hypophyseal ACTH and MSH cells were immunohistochemically characterised in the teleost fish, Diplodus sargus, using anti-ACTH (1-24) and anti alpha-MSH polyclonal antisera. ACTH cells were found both in the pars distalis and in the pars intermedia. In the former region, they appeared small, round-shaped and clustered; in the latter, they were either small or large and elongated. Moreover, a few ACTH-immunoreactive cells resembling microglia were present in the neurohypophysis. Conversely, MSH cells were found only in the pars intermedia, and were similar to the larger ACTH cells of the same region. In the pars intermedia, co-localisation of ACTH and MSH immunoreactivity in the same cell was revealed by double immunostaining, though the two hormones were also observed in distinct cell types. The distribution of ACTH cells appeared quite uniform, without any marked difference between the specimens tested. Conversely, MSH cell amount varied according to the stage of the sexual cycle of this teleost fish, which is characterised by protandrous hermaphroditism. In fact, a lower amount of MSH cells were observed in females, whereas no significant difference was found between immature and male specimens.     

Spatial patterns and feeding of meiobenthic harpacticoid copepods in relation to resident microbial flora

Distributional and feeding relationships of harpacticoid copepods and their microbial prey were examined in a tidal channel at Great Sippewissett Marsh. A horizontal zonation of photosynthetic microorganisms was composed of: 1) a diatom area; 2) a purple sulfur bacterial (Thiocapsa sp.) area; and 3) a clear area. Four species of harpacticoid copepods were associated with given areas. Leptocaris brevicornis occurred in very high densities in the diatom area but in relatively low densities in other areas. Mesochra lilljeborgi occurred in significantly higher densities in the purple and clear areas.Feeding experiments, using resident microbial flora labelled with NaH¹⁴CO₃ and ³H-thymidine, were conducted to determine which foods are 1) ingested but simply pass through the gut and 2) ingested, and retained. These experiments indicated that L. brevicornis ingested diatoms and the heterotrophs associated with the diatoms, but only retained the heterotrophic portion. Microscopic examination indicated that diatoms were passed out intact in feces. Oscillatoria sp. (cyanobacterium) was not ingested. Mesochra lilljeborgi ingested Spirulina sp. (cyanobacterium), Thiocapsa sp., and the heterotrophs associated with Thiocapsa but only retained the Thiocapsa label.These data for harpacticoids suggest that spatial distributions of meiofauna may be closely coupled with microbial food organisms which they consume. Also, that while several microbial foods may be ingested, only certain microbes are digested and assimilated as a food resource, further indicating the complexity of feeding relationships among the meiofauna.     

Feeding biology of sunfishes: patterns of variation in the feeding mechanism

Fishes are generally believed to differ in their ability to alter (modulate) their feeding behaviour in response to different prey. We investigated modulation quantitatively in four species of sunfishes (Centrarchidae) by evaluating the variation in 11 electromyographic variables measured from recordings of electrical activity in head muscles during feeding on three prey types. The experimental design used allowed us to partition variation between species, among individuals within species, among prey types, and among feedings. Duration of activity of the sternohyoideus muscle was the only variable significantly different among the four species. All variables showed significant differences among individuals within species. The overall range of activity of each muscle activity variable was about the same for all four sunfishes. However, three species showed a significant ability to modulate most muscle variables while a fourth did not change its feeding response with respect to prey type. The results indicate that: (1) intraspecific variation is an important source of variability in functional attributes that should be accounted for in comparisons between species; (2) the ability of some species to modulate appears to be independent of the potential variability in muscle activity possessed by each species; (3) closely related species can differ considerably in their ability to alter muscle electrical activity patterns during feeding; and (4) a quantitative assessment of variation in electromyographic patterns is a powerful approach for asking questions about differences in feeding behaviour.     

Biomechanical trade-offs bias rates of evolution in the feeding apparatus of fishes

Morphological diversification does not proceed evenly across the organism. Some body parts tend to evolve at higher rates than others, and these rate biases are often attributed to sexual and natural selection or to genetic constraints. We hypothesized that variation in the rates of morphological evolution among body parts could also be related to the performance consequences of the functional systems that make up the body. Specifically, we tested the widely held expectation that the rate of evolution for a trait is negatively correlated with the strength of biomechanical trade-offs to which it is exposed. We quantified the magnitude of trade-offs acting on the morphological components of three feeding-related functional systems in four radiations of teleost fishes. After accounting for differences in the rates of morphological evolution between radiations, we found that traits that contribute more to performance trade-offs tend to evolve more rapidly, contrary to the prediction. While ecological and genetic factors are known to have strong effects on rates of phenotypic evolution, this study highlights the role of the biomechanical architecture of functional systems in biasing the rates and direction of trait evolution.     

A functional approach to ecomorphological patterns of feeding in cottid fishes

Synopsis One of the major goals of an ecomorphological analysis is to correlate patterns of interspecific differences in morphology with patterns of interspecific differences in ecology. Information derived from functional morphological studies may provide a mechanistic framework supporting the correlation, but the move from a correlational relationship to a causal relationship requires experimental evidence that the interspecific morphological differences create performance differences and therefore ecological differences. The goal of this study was to examine ecomorphological relationships in the diets of cottid fishes (Scorpaeniformes: Cottidae) from the northeastern Pacific by using functional classifications of predators (based on their mode of attack) and of prey (based on their anti-predator defenses). The proposed ecomorphological relationships were tested by examining capture success among the cottids in attacks on prey with different escape abilities. As predicted on functional morphological grounds, both multivariate and univariate analyses indicated that the gravimetric importance of elusive prey (i.e. fishes, shrimp, mysids, and octopods) was greater in cottid species with a larger relative mouth size. Supporting a causal link between morphology and ecology, performance tests indicated that larger-mouthed cottids had significantly higher capture success on Heptacarpus shrimp than did smaller-mouthed species. As predicted, there were no differences in capture success among predators regardless of their attack style or ecomorphological patterns in attack on crabs (Cancer and Petrolisthes spp.) when the crabs were presented on a sand surface (i.e. easy prey). Several difficulties still exist when trying to apply a functional group approach to ecomorphology. These include the behavioral plasticity of the predators, the confounding factor of evolutionary history in identifying correlated ecomorphological features, multiple morphological solutions to common functional problems, the limitations of traditional dietary studies during extremes of prey abundance, and an inadequate understanding of the anti- predator defenses of most prey, including modifications that occur during ontogeny or in different habitats.     

The feeding behaviour of juvenile Atlantic salmon in relation to turbulent flow

The feeding behaviour of juvenile Atlantic salmon Salmo salar in the Sainte‐Marguerite River, Quebec, Canada, varied with the characteristics of turbulent flow. Simulations indicated that juveniles would decrease their swimming costs during attacks by 19·8% in low and by 31·1% in high turbulent conditions by initiating movements in low‐speed flow events. The real swimming costs did not differ from the swimming costs estimated for a situation where fish initiate their movements at randomly selected flow velocities. The juvenile Atlantic salmon did not seem to prefer low‐speed flow events when initiating their movements. The proportion of time used for movements by fish decreased with an increase in the mean and the s . d . of the flow velocity.     

An ultrastructural approach to feeding behaviour in Philodina roseola and Brachionus calyciflorus (Rotifers) I. The buccal velum

The structure situated in the lumen of the buccal tube, called here buccal velum, separates the buccal space from the pharyngeal space. The velum is a supple membrane-pile. Its structure resembles that of a fish-trap. The food, once ingested, cannot go back into the buccal lumen.