Characterization of polymorphic microsatellites in the Pacific sardine Sardinops sagax sagax (Clupeidae)

We isolated 11 microsatellites from the Pacific sardine Sardinops sagax sagax. The number of alleles and H_E among 30 individuals from a single population ranged from four to 24 and from 0.606 to 0.959, respectively. Pacific sardines are a vital economic resource in the northeastern Pacific Ocean, but insufficiently polymorphic loci have limited inferences about its stock structure and genetic variation. The level of variability of these new markers will prove useful in testing hypotheses on the stock‐structure and long‐term genetic integrity of the species.     

Prey capture and accommodation in the sandlance, Limnichthyes fasciatus (Creediidae; Teleostei)

The eyes of the sandlance, Limnichthyes fasciatus (Creediidae, Teleostei) move independently and possess a refractive cornea, a convexiclivate fovea and a non-spherical lens giving rise to a wide separation of the nodal point from the axis of rotation of the eye much like that of a chameleon. To investigate this apparent convergence of the visual optics in these phylogenetically disparate species, we examine feeding behaviour and accommodation in the sandlance with special reference to the possibility that sandlances use accommodation as a depth cue to judge strike length. Frame-by-frame analysis of over 2000 strikes show a 100% success rate. Explosive strikes are completed in 50 ms over prey distances of four body lengths. Close-up video confirms that successful strikes can be initiated monocularly (both normally and after monocular occlusion) showing that binocular cues are not necessary to judge the length of a strike. Additional means of judging prey distance may also be derived from parallax information generated by rotation of the eye as suggested for chameleons. Using photorefraction on anaesthetised sandlances, accommodative changes were induced with acetylcholine and found to range between 120 D and 180 D at a speed of 600–720 D s⁻¹. The large range of accommodation (25% of the total power) is also thought to be mediated by corneal accommodation where the contraction of a unique cornealis muscle acts to change the corneal curvatures. Accepted: 8 December 1999     

Identification of the teleost Edinger-Westphal nucleus by retrograde horseradish peroxidase labeling and by electrophysiological criteria

Summary A homolog of the Edinger-Westphal nucleus of other vertebrates is described in two species of serranid basses of the genusParalabrax, a group possessing a wide range of ocular accommodation but lacking a pupillary reflex to light. The nucleus was found by retrograde labeling from the ciliary ganglion and lies dorsolateral to the ipsilateral oculomotor nucleus. The nucleus consists of 60 to 100 neurons with an average soma diameter of about 20 m in animals weighing 70 to 150 g. Electrophysiological experiments support the identification. Microstimulation of the nucleus evokes contraction of the ipsilateral lens retractor muscle and slight constriction of the caudal ipsilateral iris. Multi- and single-unit recordings in the nucleus reveal spontaneous firing (about 30 spikes/s in single units), the rate of which decreases during visually-evoked lens retractor relaxations (accommodation to near stimuli). Recordings of muscle fiber activity in the lens retractor show essentially the same behavior, which suggests that the ciliary ganglion and neuromuscular junctions simply relay impulses with little if any synaptic integration. The existence of a discrete Edinger-Westphal nucleus devoted largely to accommodation makesParalabrax a good model system for the further tracing of central accommodation control pathways.Abbreviations CNS central nervous system - EW Edinger-Westphal nucleus - HRP horseradish peroxidase - WGA wheat germ agglutinin     

First global approach: morphological and biological variability in a genetically homogeneous population of the European pilchard, Sardina pilchardus (Walbaum, 1792) in the North Atlantic coast

The European pilchard Sardina pilchardus represents the most commercially relevant fisheries resource in many countries bordering north Atlantic coasts and the Mediterranean Sea, being especially significant along the coast of Morocco. The continuous exploitation of this pelagic species for several decades places Morocco as the leader in sardine production. However, the conditions of exploitation of this resource underwent a great change during the last recent years. In order to identify the populations of the European pilchard sardine (Sardina pilchardus, Walbaum, 1792) in the Atlantic coast of Morocco and Spain, we have combined the truss network data to conduct multivariate analysis with biologic parameters and genetic analysis based on Microsatellite and mitochondrial control region data. Sardine morphometrics data truss variables from 10 samples spanning the Atlantic coast of Morocco were analysed by multivariate analysis. Thirteen morphometric measurements and some biological parameters such as the sex and the age of fishes were made for each individual. Discriminant analysis on size-corrected truss variables and cluster analysis of mean fishes shape using landmark data indicate, that the shape of north Moroccan sardines is distinct from the shape of sardines from south Morocco. However the analysis of the mitochondrial region and four microsatellites loci (Sp2, Sp7, Sp8 and SpI5) demonstrated an homogeneity population in the Moroccan Atlantic coast, with a low but significant genetic differentiation, which followed an isolation-by-distance pattern according to Mantel test.     

Effect of maternal fat reserves on the fatty acid composition of sardine (Sardina pilchardus) oocytes

We compared the fatty acid (FA) composition of the muscle and gonads of female Iberian sardines with hydrated oocytes collected during the 2002/03 spawning season off southern Portugal (November and February) and off western Portugal (February). Sardine condition and total FA concentration in the muscle decreased between the two sampling dates, while the gonadosomatic index was similar between samples. Total monounsaturated FA concentrations in sardine gonads were different for the three samples while saturated and polyunsaturated FA concentrations were similar. Significant linear relations were found between FA concentrations in female muscle and oocytes, including eicosapentaenoic acid (EPA; 20:5n − 3) and arachidonic acid (AA; 20:4n − 6), both being essential for normal larval development. The concentration of docosahexaenoic acid (DHA; 22:6n − 3) in oocytes was independent on muscle concentration, probably resulting from its selective transfer to the oocytes. The EPA/DHA ratio was highly conserved in sardine tissues, while DHA/AA and EPA/AA ratios varied significantly between samples. These results indicate that the FA content of eggs produced by sardines varies throughout the spawning season, egg FA concentrations decreasing as females lose condition, and FA composition also shows spatial variability. Both types of variability may have a significant impact on egg quality, particularly on the amount of reserves available to larvae affecting their resistance to starvation, and the appropriate FA composition required for normal growth.     

Multivariate analyses of the biometric features from Philippine sardines – implications for the phylogenetic relationships of the freshwater Sardinella tawilis (Teleostei,Clupeomorpha)*

Philippine sardines are members of the commercially important Sardinella genus (Family Clupeidae). Sardines are mostly marine species with a very few exception like the Philippine freshwater sardine, Sardinella tawilis, of Lake Taal. This species is believed to have immigrated from Balayan Bay some 250 years ago. To determine the relationship of freshwater S. tawilis to the marine forms, 35 biometric features were investigated. The variables were subjected to univariate (mean, standard deviation and coefficient of variance) and multivariate analyses [Principal Component Analysis (PCA) and Hierarchical Cluster Analysis]. Results from univariate tests revealed that the number of ventral fin ray can discriminate Sardinella lemuru from the rest of the sardines. Multivariate PCA, on the other hand, showed discrete separation of the marine sardines. Sardinella tawilis, however, cannot be clearly separated from the marine species, Sardinella albella. The same clustering was recovered in HCA, thus providing evidence that S. albella is the closest marine relative of the freshwater S. tawilis.     

Eyes and vision in Arion rufus and Deroceras agreste (Mollusca; Gastropoda; Pulmonata): What role does photoreception play in the orientation of these terrestrial slugs?

This paper deals with the orientational behaviour in the two terrestrial slugs Arion rufus and Deroceras agreste. It presents anatomical details of their eyes and provides an appraisal of the eyes’ optical system. In both species the retinae contain two principal types of cell: photoreceptive and pigmented supportive cells. While only the eye of A. rufus apparently contains neurosecretory neurones, that of D. agreste is the only one equipped with a small additional retina with its own separate lens. Lens shapes vary between ovoid (A. rufus) and spherical (D. agreste). Our results demonstrate that the camera‐type eyes in A. rufus and D. agreste have optical systems that do not allow the production of a sharp image on the retina. The slugs demonstrate negative visually mediated phototactic behaviour, but no polarization sensitivity. Only one aspect of the visual environment, namely the overall distribution of light and dark, seems to be important for these slugs. As the main role of the slugs’ photoreceptors is to monitor environmental brightness and to assist the animal in orientating towards dark places, we conclude that these slugs do not need to perceive sharp images.     

18S rRNA V9 metabarcoding for diet characterization: a critical evaluation with two sympatric zooplanktivorous fish species

The potential of the 18S rRNA V9 metabarcoding approach for diet assessment was explored using MiSeq paired‐end (PE; 2 × 150 bp) technology. To critically evaluate the method′s performance with degraded/digested DNA, the diets of two zooplanktivorous fish species from the Bay of Biscay, European sardine (Sardina pilchardus) and European sprat (Sprattus sprattus), were analysed. The taxonomic resolution and quantitative potential of the 18S V9 metabarcoding was first assessed both in silico and with mock and field plankton samples. Our method was capable of discriminating species within the reference database in a reliable way providing there was at least one variable position in the 18S V9 region. Furthermore, it successfully discriminated diet between both fish species, including habitat and diel differences among sardines, overcoming some of the limitations of traditional visual‐based diet analysis methods. The high sensitivity and semi‐quantitative nature of the 18S V9 metabarcoding approach was supported by both visual microscopy and qPCR‐based results. This molecular approach provides an alternative cost and time effective tool for food‐web analysis.     

The Fine Structure of the Compound Eyes of Shallow-Water Asellotes,Jaera albifrons Leach and Asellus aquaticus L. (Crustacea: Isopoda)

Abstract Both species have small sessile compound eyes. The dioptric apparatus of J. albifrons consists of a biconvex lens and a pyriform crystalline cone, the latter formed by two principal and two accessory cone cells. A. aquaticus has a reduced lens and a round cone formed by two to four principal cone cells with two to no accessory cone cells. Distal pigment cells and pigmented retinular cells lie between the ommatidia in J. albifrons. A. aquaticus has only the pigmented retinular cells. Both species have a fused, continuous (unhanded) rhabdom formed by eight retinular cells (R1—8), one of which (R8) is situated distally. The retinular cells R1—7 form, in J. albifrons, a cylinder-shaped middle portion with three microvillar directions (60° apart) and a proximal star-shaped portion. The entire rhabdom of A. aquaticus is star-shaped. Distal pigment-cell processes and basal cells form the fenestrated membrane in J. albifrons and “eye-cup cells” in A. aquaticus.     

Structure and development of the larval visual system in embryos of Lytta viridana leconte (coleoptera,meloidae)

At hatching (252–264 hr. at 25 ± 0.5°C), the visual system in larvae of Lytta viridana consists of paired stemmata, stemmatal nerves, optic neuropiles, and inner and outer imaginal optic lobe anlagen. It originates between 64 and 72 hr. with invagination of an optic lobe primordium in the side of each protocephalic lobe. These primordia later differentiate into protocerebral ganglion cells and the imaginal optic lobe anlagen. Each stemma arises at 72 hr. from epidermis below and behind the optic lobe invagination and subsequently becomes cupshaped, closes over, and differentiates. At hatching, it consists of a planoconvex corneal lens, a corneagenous layer, and an everse retina of numerous, pigmented retinular cells, each with a terminal rhabdomere. Between 96 and 104 hr, proximal ends of the retinular cells grow posteromedially into a transverse, horizontal fold in the posterior wall of each optic lobe invagination and along its length to the protocerebral neuropile, which they contact by 112 hr. As the brain withdraws posteriorly within the head, these axons elongate correspondingly. Sheath cells of stemmata and stemmatal nerves descend either from protocerebral perineurium or the optic lobe primordia. Structure and development of the larval visual system in L. viridana are compared with those of other insects and its various components are shown to be homologous throughout the Insecta. However, the stemmata of this insect more closely resemble the atypical imaginal eyes of male scale insects than the photoreceptors of other holometabolous larvae–a similarity arising through convergence.     

Ultrastructure of prostomial photoreceptors in four marine polychaete species (annelida)

The photoreceptors of four polychaete species were investigated by transmission electron microscopy: Eteone longa and Anaitides mucosa (Phyllodocidae), Scolelepis squamata (Spionidae), and Heteromastus filiformis (Capitellidae). Four different types of light-sensitive organs could be distinguished: 1) a simple, unpigmented rhabdomeric type; 2) a simple ocellus composed of a sensory and a pigmented cell; 3) complex eyes with a lens consisting of secretory granules; 4) a simple, unpigmented type with modified cilia. In spite of its simpler organization the fourth type is listed last, because its function as a photoreceptor seems dubious. The first type (unpigmented rhabdomeric receptor) occurs in all four species investigated. It is the only type of photoreceptor in Heteromastus. Additionally, the two phyllodocids Eteone and Anaitides possess another kind of receptor (type 4) in close proximity to the type 1 receptor. Simple ocelli (type 2) are found in Scolelepis. A pair of complex eyes (type 3) is present in both Eteone and Anaitides, but they show important differences in the two species. First, the eyes in Eteone exhibit ciliary rudiments within the sensory processes, but such rudiments are absent in the eyes of Anaitides. Secondly, the sensory cells in Anaitides possess pigment granules, whereas in Eteone they do not. Thirdly, the lens in Eteone is composed of secretion granules of equal electron density, whereas in Anaitides the lens granules show increased electron density centrally. Lens material appears to be secreted from a single corneal cell in Eteone, and from several corneal cells in Anaitides. In both species these corneal cells are located distally outside the lens.     

Development of a unique eye: photoreceptors of the pelagic predator Atlanta peroni (Gastropoda, Heteropoda)

 The eyes of different larval stages and juveniles of Atlanta peroni are generally composed of a cornea, a lens and a retina. In juveniles a distinct pigmented shield is visible and an enormous humour is located behind the lens. Larvae have only two sensory cells and the photoreceptors are of the ciliary type. In juveniles a striking feature is the shape of the retina. It is ribbon-shaped and new sensory cells are present which are arranged in three rows. The photoreceptors are of the ciliary type as well. Contrary to the arrangement in larvae, the ciliary plasmalemma in juveniles forms numerous lamellar stacks. In accordance with the sensory cells the stacks are organized in three parallel rows. The lamellae of adjacent stacks within a row overlap each other. The latter unique feature has not yet been found in any other representative of the Heteropoda. These findings demonstrate that (a) the eyes are altered during the development from larvae into juveniles, (b) the larval sensory cells are reduced and replaced by new sensory cells in juveniles and (c) the eyes of juvenile and adult A. peroni are well adapted for their life as visual predators. Accepted: 20 February 1999     

Amyloodiniosis in farmed Sardinella brasiliensis (Steindachner, 1879): Case report of successful therapeutic control with copper sulfate

Brazilian sardine is emerging as a promising species in Aquaculture. This study describes for the first time a case of parasitic infestation by Amyloodinium in Brazilian sardines Sardinella brasiliensis obtained from natural spawning in captivity. The sardines kept in nurseries were naturally parasitized by the amylodiniosis causative agente the dynoflagellate A. ocellatum with high mortalities above 50%. Fish presented clinical signs characteristic of amyloodiniosis which included easily perceived behavioral changes such as loss of appetite, scraping of the body against objects, walls and bottom, nursery pipes, agglomerations near the aerators and water inlets, presented with accelerated opercular beating and erratic swimming. For therapeutic treatment copper sulfate was used for 10 days. At the end of the treatment period the fish had no clinical signs or presence of the parasite on the body surface, indicating that the application of copper sulfate in concentration of 0.2 mg L⁻¹ of Cu⁺ was effective to control this parasite, apparently without causing damage to Brazilian sardine.     

Compound eyes and ocular pigments of crustacean larvae (Stomatopoda and decapoda,brachyura)

Larvae of decapod and stomatopod crustaceans possess paired compound eyes not unlike those of adult crustaceans. However, the visual demands of larval and adult life differ considerably. Furthermore, the eyes of adult stomatopods appear to be far more specialized than those of the larvae. We examined eyes of several stomatopod species just before and after larval metamorphosis. At this time, the entire larval retina is joined by a new, adult‐type retinal array which gradually replaces the remnants of the larval retina. The new retina of the postlarva is anatomically similar to that of the full‐grown adult, and has virtually identical assemblages of intrarhabdomal filters. We determined the photopigments of Gonodactylus aloha, the only species for which we were able to obtain both larval and adult specimens, using microspectrophotometry. The single middle‐wavelength larval rhodopsin (λ_max= 499 nm) disappears at metamorphosis; none of the 10 classes of adult rhodopsins has λ_max between 473 and 510 nm. This metamorphic change of visual pigment does not occur in a comparison species of decapod crustacean, the blue crab Callinectes sapidus. Here, rhodopsins both of the megalops larva and the adult had λ_max at 503–504 nm. The difference between these two species can be explained by the varying ecological requirements of their larvae and adults, and more study of visual pigments in retinas of larval and adult crustaceans is warranted.     

Chemical characteristics and lipid fraction quality of sardines (Sardina pilchardus W.): influence of sex and length

An experimental investigation was carried out on sardines (Sardina pilchardus W.) with the aim to evaluate the influence of sex and total length (TL) on total weight (TW) and fillet weight (FW), chemical composition, and lipid fraction quality. Special emphasis was given with respect to the content of polyunsaturated fatty acids (PUFA), which are considered important from a dietary, nutritional, and functional point of view. Sardines were subdivided as a function of TL and sex. Four groups were obtained: (i) males, TL 130–155 mm, (ii) males, TL 155–180 mm, (iii) females, TL 130–155 mm, (iv) females, TL 155–180 mm. Fatty acid composition and chemical analyses were subsequently carried out on the sardine fillets; results showed higher TW and FW in female sardines and higher percentage value of FW/TW ratio in male sardines, indicating a higher incidence of viscera and other non‐edible parts in females than in males. Moreover, total lipids were on average significantly higher (P < 0.05) in longer specimens (TL = 155–180 mm) than in shorter ones (TL = 130–155 mm). Finally, total saturated fatty acids (SFA) and total mono‐unsaturated fatty acids (MUFA) values were found to be significantly higher in females than in males; such values increased significantly (P < 0.01) only for total SFA from shorter to longer sardines. As for total PUFA, both ω6 and ω3 were found to be higher in male than in female sardines and significantly higher (P < 0.001) in larger than in smaller sizes. Total PUFA/total SFA ratio were in all cases close to 1, with higher values in male than in female sardines and in larger than in smaller specimens.     

Comparative morphological studies on the visual systems in a binocular and a multi-ocular species of freshwater planarian

The visual systems of Bdellocephala brunnea Ijima & Kaburaki, a species with two eyes, and Polycelis sapporo (Ijima & Kaburaki), a species with multiple eyes, were investigated using light and electron microscopy. The eye of the binocular species consisted of 40–50 photoreceptor cells and 6–12 pigmented eyecup cells. The eye of the multi-ocular species was smaller and consisted in most specimens of one photoreceptor cell and one pigmented eyecup cell. The ultrastructure of the photoreceptor cells and of the pigmented cells was similar in the two species. Despite differences in numbers of constitutive cells, the arrangement of functional elements in the ocelli of these planarians is the same.     

Mitochondrial DNA Sequence Variation Suggests the Lack of Genetic Heterogeneity in the Adriatic and Ionian Stocks of Sardina pilchardus

A genetic stock structure analysis of 11 sardine samples from the Adriatic Sea and Ionian neighboring area was carried out through sequence variation analysis of a 307-bp cytochrome b gene fragment in order to identify self-recruiting units in the Adriatic Sardina pilchardus stock. The overall lack of genetic subdivision among samples detected by analysis of molecular variance, pairwise Φ_st values, and the exact test of population differentiation indicates this sardine stock is part of a larger self-recruiting population whose boundaries are larger than the investigated area. This conclusion is in agreement with preliminary allozymic and mitochondrial DNA restriction fragment length polymorphism data, but contradicts the previous identification of 2 subpopulations of sardines in the Adriatic Sea argued on morphologic differences, which could be rather attributed to different hydrographic or ecologic conditions occurring in different areas of the Adriatic Sea. The reduced gene flow observed between Adriatic-Ionian and Spanish sardine geographic samples (P < 0.001) suggests that reproductively isolated populations of sardines may occur in the Mediterranean Sea.     

Pigmented eyes,photoreceptor‐like sense organs and central nervous system in the polychaete Scoloplos armiger (Orbiniidae,Annelida) and their phylogenetic importance

The phylogenetic position of Orbiniidae within Annelida is unresolved. Conflicting hypotheses place them either in a basal taxon Scolecida, close to Spionida, or in a basal position in Aciculata. Because Aciculata have a specific type of eye, the photoreceptive organs in the orbiniid Scoloplos armiger were investigated to test these phylogenetic hypotheses. Two different types of prostomial photoreceptor‐like sense organs were found in juveniles and one additional in subadults. In juveniles there are four ciliary photoreceptor‐like phaosomes with unbranched cilia and two pigmented eyes. The paired pigmented eyes lie beside the brain above the circumoesophageal connectives. Each consists of one pigmented cell, one unpigmented supportive cell and three everse rhabdomeric sensory cells with vestigial cilia. During development the number of phaosomes increases considerably and numerous unpigmented sense organs appear consisting of one rhabdomeric photoreceptor cell and one supportive cell. The development and morphology of the pigmented eyes of S. armiger suggest that they represent miniaturized eyes of the phyllodocidan type of adult eye rather than persisting larval eyes resulting in small inverse eyes typical of Scolecida. Moreover, the structure of the brain indicates a loss of the palps. Hence, a closer relationship of Orbiniidae to Phyllodocida is indicated. Due to a still extensive lack of ultrastructural data among polychaetes this conclusion cannot be corroborated by considering the structure of the unpigmented ciliary and rhabdomeric photoreceptor‐like sense organs. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.     

Functional morphology of the visual system and mating strategies in bumblebees (Hymenoptera,Apidae, Bombus)

Bumblebee mating behaviour intrigued Darwin over 150 years ago and has elicited much interest since. Despite our increasing knowledge about male mating strategies, we still know little about the signals and cues involved, the sender‐receiver relationship and the functional adaptations of the involved sensory systems. Although several authors have noticed a close relationship between the mating system and male‐specific elaboration of the visual system, studies on the functional adaptations have yet to be conducted. We investigated the functional morphology of the compound eyes in 11 species of Bombus from various subgenera in detail. Of these, four species (Bombus confusus, Bombus melaleucus, Bombus mendax, Bombus niveatus) have enlarged male eyes, characterized by a higher number of ommatidia compared with worker eyes and a frontal zone with enlarged facets, which is likely to be associated with improved spatial resolution and contrast sensitivity. In these species, perching mating strategies are found. In contrast, males of species that patrol scent routes (Bombus hortorum, Bombus lapidarius, Bombus pascuorum, Bombus pratorum, Bombus soroeensis, Bombus terrestris, Bombus wurflenii) show no distinct eye adaptations, and their eye morphology closely resembles that of the workers. A phylogenetic analysis of male eye structure indicates that enlarged males eyes may have been the ancestral state in Bombus. © 2014 The Linnean Society of London