Journal of Fish Biology

Three normalized cDNA libraries were constructed, two of which were constructed from reproductive tissues ovary and testis, and the other one from pooled immune tissues including head kidney, intestine, liver and spleen. A total of 10 542 clones were sequenced generating 10 128 expressed sequence tags (ESTs). Cluster analysis indicated a total of 5808 unique sequences including 1712 contigs and 4096 singletons. A total of 4249 (73%) of the unique ESTs had significant hits to the non-redundant protein database, 2253 of which were annotated using Gene Ontology (GO) terms. A total of 311 microsatellites (with 246 having sufficient flanking sequences for primer design) and 6294 putative SNPs were identified. These genome resources provide the material basis for future microarray development, marker validation and genetic linkage and QTL analysis.

Late summer sampling of pelagic age-0 year fish communities in five Czech reservoirs and one Dutch reservoir revealed extremely small age-0 year pikeperch Sander lucioperca (mean 24 mm standard length, LS , minimum 13 mm LS ) alongside more normal-sized S. lucioperca that are found at the end of the first growing season (mean 50 mm LS ), resulting in two clearly size-separated cohorts. Reference to such small age-0 year S. lucioperca in lakes or reservoirs at this time of year and in such large numbers are almost absent the scientific literature, and the presence of these small S. lucioperca is contradictory to the common understanding of the reproductive biology of this species. This overlooked phenomenon may have a major effect on the population dynamics of this valuable species because of size-dependent winter mortality.

Eleven novel polymorphic microsatellite loci were developed and characterized for the recently validated roundscale spearfish Tetrapturus georgii. Characterization of these markers, based on 35 roundscale spearfish from the western North Atlantic, revealed two to 21 alleles per locus with an average expected heterozygosity (H(E) ) of 0·09-0·94, and all loci conformed to Hardy-Weinberg expectations. Cross-amplification of these 11 loci against all other eight known istiophorid species indicates promising prospects for the utility of these markers for istiophorids in general.

A collection of fishes from the Brazilian continental slope between 11 degrees and 23 degrees S obtained through trawling revealed nine species of Ipnopidae. Bathypterois bigelowi and Bathytyphlops marionae represent first records from the south-western Atlantic Ocean and Bathypterois grallator is reported off Brazil for the first time. Four species have their distribution extended in Brazilian waters: Bathypterois phenax, Bathypterois quadrifilis, Bathypterois viridensis and Ipnops murrayi. An identification key of Ipnopidae species from the south-western Atlantic Ocean is included.

Commercially available enzyme immunoassay (EIA) kits were validated for measuring steroid hormone concentrations in blood plasma from three fish species: the orange clownfish Amphiprion percula, the orangefin anemonefish Amphiprion chrysopterus and the blacktip reef shark Carcharhinus melanopterus. A minimum of 5 microl plasma was required to estimate hormone concentrations with both kits. These EIA kits are a simple method requiring minimal equipment, for measuring hormone profiles under field conditions.

The relationship between whole-body concentrations of 11-ketotestosterone (11-KT) and sexual function was examined in the coral goby Gobiodon erythrospilus, a bi-directional sex-changing fish. 11-KT occurred in both female and male G. erythrospilus, but levels were not always higher in males than in females within heterosexual pairs, and were not related to the stage of gonadal development of individual fish. These results suggest that comparable 11-KT levels in both sexes may allow serial adult sex change to take place in bi-directional sex-changing species, such as Gobiodon spp.

This study details 13 novel polymorphic microsatellite loci in the armoured catfish Hypostomus ancistroides, and assesses their utility for population genetic studies. The analysis of 30 individuals revealed a total of 99 different alleles (ranging from two to 15 alleles per locus), with an average of 7·62 alleles per locus, with observed and expected heterozygosities ranging from 0·103 to 0·931 and from 0·102 to 0·906, respectively. One of the 13 loci showed significant deviations from Hardy-Weinberg equilibrium, probably due to the presence of null alleles, inferred from the excess of homozygotes. © 2015 The Fisheries Society of the British Isles.

Hydrostatic pressure elevated to 500 kPa for 14 days was found to affect hepatic 7-ethoxyresorufin-O-deethylase (EROD), oxidized protein (POx), protein yield and branchial Na(+)-K(+)-ATPase. No effect on glutathione-S-transferase (GST), superoxidase dismutase (SOD), catalase (CAT), lipid peroxidation (LP), acetylcholinesterase (AChE), butyrylcholinesterase (BChE), condition factor (K) and hepato-somatic index (I(H)) was encountered.

Fourteen polymorphic microsatellite loci were isolated and characterized for the Neotropical cichlid Geophagus brasiliensis and tested on 30 individuals belonging to a single population. Among the 14 loci described, four showed potential presence of null alleles, inferred from the excess of homozygous genotypes, and three of these loci showed significant deviations from Hardy-Weinberg equilibrium. Fifty-nine different alleles were detected (ranging from two to eight alleles per locus), with estimates of observed and expected heterozygosity ranging from 0·167 to 0·700 and from 0·269 to 0·825. Cross-amplification of primers was successful in five other cichlid species.

In this study, although the highest production of two physiologically significant progestins in teleosts [17,20β-dihydroxypregn-4-en-3-one (17,20β-P) and 17,20β,21-trihydroxypregn-4-en-3-one (17,20β,21-P)] was observed in the period just prior to spawning in both male and female roach Rutilus rutilus, there was also a substantial production (mean levels of 5-10 ng ml(-1) in blood; and a rate of release of 5-20 ng fish(-1) h(-1) into the water) in males and females in the late summer and early autumn (at least 7 months prior to spawning). During this period, the ovaries were increasing rapidly in size and histological sections were dominated by oocytes in the secondary growth phase [i.e. incorporation of vitellogenin (VTG)]. At the same time, the testes were also increasing rapidly in size and histological sections were dominated by cysts containing mainly spermatogonia type B. Measurements were also made of 11-ketotestosterone (11-KT) in males and 17β-oestradiol and VTG in females. The 3 months with the highest production of 11-KT coincided with the period that spermatozoa were present in the testes. In females, the first sign of a rise in 17β-oestradiol concentrations coincided with the time of the first appearance of yolk globules in the oocytes (in August). The role of the progestins during the late summer and autumn has not been established.

Atlantic cod Gadus morhua ovaries were incubated in vitro with tritiated 17-hydroxypregn-4-ene-3,20-dione (17-P) to determine whether 17,20beta-dihydroxypregn-4-en-3-one (17,20beta-P) or 17,20beta, 21-trihydroxypregn-4-en-3-one (17,20beta,21-P), or both, were more likely to be the steroid responsible for inducing oocyte final maturation (i.e. resumption of meiosis). Only 17,20beta,21-P was produced, in addition to 11-deoxycortisol (17,21-P), which is intermediate between 17-P and 17,20beta,21-P. Also, the 5beta-reduced forms of 17-P, 17,21-P and 17,20beta,21-P were all found. Some sulphation of 21-hydroxylated steroids was demonstrated. The ability of female G. morhua to make 17,20beta,21-P but not 17,20beta-P was confirmed by radioimmunoassay of plasma samples from spawning fish. Although small amounts of 17,20beta-P immunoreactivity were detected in a few plasma samples, this was shown, by thin-layer chromatography, to be mostly due to cross-reaction with other unidentified compounds. The evidence strongly suggests that 17,20beta,21-P is more likely than 17,20beta-P to be the maturation-inducing steroid in G. morhua.

The major progestin in teleosts is not progesterone, as in tetrapods, but 17,20beta-dihydroxypregn-4-en-3-one (17,20beta-P) or, in certain species, 17,20beta,21-trihydroxy-pregn-4-en-3-one (17,20beta,21-P). Several functions for 17,20beta-P and 17,20beta,21-P have been proposed (and in some cases proved). These include induction of oocyte final maturation and spermiation (milt production), enhancement of sperm motility (by alteration of the pH and fluidity of the seminal fluid) and acting as a pheromone in male cyprinids. Another important function, initiation of meiosis (the first step in both spermatogenesis and oogenesis), has only very recently been proposed. This is a process that takes place at puberty in all fishes and once a year in repeat spawners. The present review critically examines the evidence to support the proposed functions of 17,20beta-P in males, including listing of the evidence for the presence of 17,20beta-P in the blood plasma of male fishes and discussion of why, in many species, it appears to be absent (or present at low and, in some cases, unvarying concentrations); consideration of the evidence, obtained mainly from in vitro studies, for this steroid being predominantly produced by the testis, for its production being under the control of luteinizing hormone (gonadotrophin II) and, at least in salmonids, for two cell types (Leydig cells and sperm cells) being involved in its synthesis; discussion of the factors involved in the regulation of the switch from androgen to 17,20beta-P production that seems to occur in many species just at the time of spermiation; discussion of the effects of in vivo injection and application of 17,20beta-P (and closely related compounds) in males; a listing of previously published evidence that supports the proposed new function of 17,20beta-P as an initiator of meiosis; finally, discussion of the evidence for environmental endocrine disruption by progestins in fishes.

The effect of extended incubation (delayed hatching) on larval morphology in the terrestrially spawning common galaxias Galaxias maculatus was investigated by inducing larvae to hatch 1 and 2 weeks after the normal 2 week incubation period. After 1 week of extended incubation, larvae were larger (longer in standard length, L(S), and greater in body depth) compared to controls (larvae that experienced normal incubation durations). After 2 weeks of extended incubation, larvae were smaller (shorter in L(S) and smaller in body depth) than larvae that experienced 1 week of extended incubation. Furthermore, eye area increased while yolk-sac size decreased monotonically with increasing incubation duration. These results suggest that larvae experiencing long periods of extended incubation are using somatic tissue to meet their metabolic demands. Larvae that experienced 2 weeks of extended incubation succumbed to starvation sooner than control larvae, but hatching success was not significantly different. Temperature mediated the effect of extended incubation on the morphology of larvae at hatching, most likely, through its effects on developmental rate and efficiency of yolk utilization. This study demonstrates some of the consequences of terrestrial spawning with extended incubation, which will assist in determining why this intriguing behaviour has evolved several times in a diverse range of taxa.

A revision of fish specimens previously identified as Heptapterus mustelinus from the endorheic Río Salí Basin, Tucumán, Argentina, reveals that they present several morphological differences from that species. This paper describes Heptapterus qenqo sp. nov. from the Río Salí Basin. The new species is diagnosed by a combination of the following characters: presence of small serrae on the anterior proximal margin of the first pectoral-fin ray; anal-fin rays iv-v, 11-13 (15-17 total anal-fin rays); adipose-fin base 40·9-47·4% standard length; small eyes (7·4-14·2% head length); adipose-fin confluent to caudal fin and maxillary barbel not reaching pectoral-fin base in adults, and reaching or scarcely surpassing the first pectoral-fin ray in small juveniles.

Two new species of Leporinus are described from tributaries of the Rio Amazonas in Brazil. One species is known from the Jari and Tapajós River basins, and is identified on the basis of a gas bladder reduced in size, a dark midlateral stripe on the body, dark transverse bars on the dorsum, a subinferior mouth, three teeth on the premaxilla, four teeth on the dentary and 16 scale rows around the caudal peduncle. The second new species is known from the Tocantins, Xingu and Tapajós River basins, and is identified on the basis of three dark longitudinal stripes on the body, a subinferior mouth, three teeth on the premaxilla, four teeth on the dentary and 12 scale rows around the caudal peduncle. In addition, Leporinus striatus is redescribed based on type and additional specimens from the Río de La Plata, Amazonas, Orinoco, Atrato, Magdalena and Sinu River basins. Leporinus striatus is identified on the basis of four dark longitudinal stripes on the body, a subterminal mouth, three teeth on the premaxilla, four teeth on the dentary and 16 scale rows around the caudal peduncle.

Three species of cichlids belonging to the genus Symphysodon have demonstrated interspecific and intraspecific variation in nucleolus organizer regions (NOR) detected with silver nitrate. In order to understand the evolution of this marker in the genus, the structural variability of these sequences in mitotic chromosomes from Symphysodon aequifasciatus, Symphysodon discus and Symphysodon haraldi was investigated using both silver nitrate impregnation and hybridization of the 18S rRNA gene probe. For the three species, the two markers were intraspecifically and interspecifically variable both in the number and in the size of the sites. This polymorphism may stem from duplications and translocations, which suggests that structural chromosome rearrangements effectively act in the karyoevolution of wild Symphysodon species and may have favoured the adaptability of these fishes to diverse aquatic environments in the Amazon.

The fractionation of an aqueous extract of yam Dioscorea antaly from Madagascar led to the isolation of terpenoids and flavonoids. Compounds were identified on the basis of modern mass spectrometry and two-dimensional nuclear magnetic resonance (2D-NMR). Toxicological effects of the most abundant isolated compound, 8-epidiosbulbin E were studied on medaka Oryzias latipes embryo-larval development. The lethal concentration (killing 50%; LC(50) ) to embryos treated 24 h before hatching and for 3 days after hatching was estimated to be 0·56 mg ml(-1) (P< 0·05). No mortality was observed with O. latipes larvae exposed after hatching until day 4. Anatomo-pathological studies of embryos exposed to 0·56 mg ml(-1) showed development anomalies of the central nervous system, liver, muscle and intestine. The present data thus extend the model of O. latipes embryos as a useful animal model to analyse the effects of food toxins.

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This study tested the 'silver spoon' hypothesis which posits that individuals that develop under favourable conditions should enjoy a fitness advantage later in life because they are more likely to recognize and settle in high-quality habitats. Atlantic salmon Salmo salar of two age classes (0+ and 1+ years) were reared in environmentally enriched or standard hatchery tanks for a short period (c. 10 weeks), were then released into a natural river and sampled on repeated occasions to test for silver-spoon effects. Compared with controls, enriched fish had a 6·4% higher recapture rate and settled in higher velocity habitats when they were stocked as 0+ year fry, but not when they were stocked as 1+ year parr. The opportunity for selection was generally higher for environmentally enriched fish than for controls, and also higher for 0+ than for 1+ year fish. Selection favoured individuals with high condition factor, extensive fat reserves and longer than average pectoral fins in both age classes but favoured a small body size in 1+ year and a large body size in 0+ year releases. Stomach analysis showed that enriched fish ate more, and adapted quicker to natural prey than controls. These results provide support for silver-spoon effects in fish and indicate that enrichment can improve post-release performance in conservation programmes, but seemingly only if fish are not kept in captivity for too long. © 2014 The Fisheries Society of the British Isles.

To investigate Brazilian freshwater ichthyology, from 1986 to 2005, a bibliometric analysis was conducted using abstracts downloaded from The Web of Science database searching for the keywords 'fish', 'pisces', 'teleostei' and the address field having the word 'Brazil'. The results of this study showed that Brazilian freshwater ichthyology publications have been increasing during the study period. This process is a consequence of a series of investments that the Brazilian Government has made. Furthermore, data analyses identified scientific areas where there was a lack of scientific knowledge (e.g. studies of species threatened with extinction and certain hydrologic basins). Research institutions from the north-east and northern region of Brazil had the lowest participation in scientific productivity, which was a reflection of their regions poorer economic situation. This study showed that scientific productivity in Brazilian ichthyology was a direct reflection of state investment in research. Furthermore, data in this study follow expected statistical probabilities, for example, fishes from the most diverse families were the most studied. Thus, the study shows that great progress has been made by Brazilian ichthyologists in the last 20 years; however, due to the mega diversity of fishes in Brazil, much remains to be done if many species are to become known to science and to be saved from extinction. This it seems will depend on continued and further investment by Brazilian Government funding agencies, as Brazilian ichthyologists have demonstrated their capacity to generate high quality information about their study species.

Census (N(C)) and effective population size (N(e)) were estimated for a lake-resident population of brown trout Salmo trutta as 576 and 63, respectively. The point estimate of the ratio of effective to census population size (N(e):N(C)) for this population is 0.11 with a range of 0.06-0.26, suggesting that N(e):N(C) ratio for lake-resident populations agree more w(i)th estimates for fishes with anadromous life histories than the small ratios observed in many marine fishes. (C) 2011 The Authors Journal of Fish Biology (C) 2011 The Fisheries Society of the British Isles

The effect of temperature and mass on specific growth rate (G) was examined in spotted wolffish Anarhichas minor of different size classes (ranging from 60 to 1500 g) acclimated at different temperatures (4, 8 and 12 degrees C). The relationship between G and 20S proteasome activity in heart ventricle, liver and white muscle tissue was then assessed in fish acclimated at 4 and 12 degrees C to determine if protein degradation via the proteasome pathway could be imposing a limitation on somatic growth. Cardiac 20S proteasome activity was not affected by acclimation temperature nor fish mass and had no correlation with G. Hepatic 20S proteasome activity was higher at 12 degrees C but did not show any relationship with G. Partial correlation analysis showed that white muscle 20S proteasome activity was negatively correlated to G (partial Pearson's r = -0.609) but only at cold acclimation temperature (4 degrees C). It is suggested that acclimation to cold temperature involves compensation of the mitochondrial oxidative capacity which would in turn lead to increased production of oxidatively damaged proteins that are degraded by the proteasome pathway and ultimately negatively affects G at cold temperature.

This review identifies a number of exciting new developments in the understanding of vision in cartilaginous fishes that have been made since the turn of the century. These include the results of studies on various aspects of the visual system including eye size, visual fields, eye design and the optical system, retinal topography and spatial resolving power, visual pigments, spectral sensitivity and the potential for colour vision. A number of these studies have covered a broad range of species, thereby providing valuable information on how the visual systems of these fishes are adapted to different environmental conditions. For example, oceanic and deep-sea sharks have the largest eyes amongst elasmobranchs and presumably rely more heavily on vision than coastal and benthic species, while interspecific variation in the ratio of rod and cone photoreceptors, the topographic distribution of the photoreceptors and retinal ganglion cells in the retina and the spatial resolving power of the eye all appear to be closely related to differences in habitat and lifestyle. Multiple, spectrally distinct cone photoreceptor visual pigments have been found in some batoid species, raising the possibility that at least some elasmobranchs are capable of seeing colour, and there is some evidence that multiple cone visual pigments may also be present in holocephalans. In contrast, sharks appear to have only one cone visual pigment. There is evidence that ontogenetic changes in the visual system, such as changes in the spectral transmission properties of the lens, lens shape, focal ratio, visual pigments and spatial resolving power, allow elasmobranchs to adapt to environmental changes imposed by habitat shifts and niche expansion. There are, however, many aspects of vision in these fishes that are not well understood, particularly in the holocephalans. Therefore, this review also serves to highlight and stimulate new research in areas that still require significant attention.

The interspawning interval, or spawning frequency, of wild three-spined stickleback, Gasterosteus aculeatus, was estimated using histological examination of postovulatory follicles (POF). Females in Alaskan lakes appeared to have as much as a 48 h delay between ovulation and ovoposition, yet the POF method could still be used to estimate the interspawning interval. In two Alaskan lakes the interspawning interval was estimated to range from 2.2 to 7.8 days among individual female G. aculeatus. These estimates were consistent with the range (2.5 to 5 days) of previous estimates among individual females from laboratory observations of spawning G. aculeatus, as well as anecdotal accounts of spawning intervals reported from wild populations in Canada (5-10 days). The interspawning interval of females increased during the course of the spawning season in Alaska, showing that the majority of female spawning activity occurred during the earliest portion of the approximate 6-week reproductive season. The increased interspawning interval appears to be related to a previously reported decrease in body condition in reproductive females during the breeding season. Thus, female G. aculeatus may be unable to sustain the initial rate of reproduction as energy stores that support the rapid growth of vitellogenic oocytes are depleted.

Temperature and egg viability data from an Arctic charr Salvelinus alpinus hatchery covering a period of 28 years were analysed. During the study period, there was a significant increase in the mean water temperature in May, July, August and September of c. 2° C. Independent of year, the egg viability showed a negative correlation with the mean monthly temperatures in July, August and September as well as with the temperature difference between October and November. The negative effect of high summer temperatures was further supported by a comparison of egg viability from replicate broodstock reared at two sites differing mainly in summer water temperature. The eggs from the colder site were, on average, significantly larger (4·4 mm compared with 4·0 mm) and had higher hatching rates (57% compared with 37%). These results suggest that unfavourable temperature conditions during the summer and autumn can explain much of the excessive egg mortality experienced at the main facility used for the Swedish S. alpinus breeding programme. The main effect was supra-optimal temperatures during the period July to September, but there also appears to have been an effect from the temperature regime before and during spawning (October to November) that was unrelated to the summer temperatures. These findings emphasize the importance of site selection and sustainable management of aquaculture hatcheries in the light of the ongoing climate change. © 2015 The Fisheries Society of the British Isles.

A method for oxygenating and mixing suspensions of turbot Psetta maxima red blood cells (RBC) was tested in (31)P nuclear magnetic resonance (NMR) spectroscopy. In normoxia, the levels of inorganic phosphate (Pi) and nucleoside triphosphates (NTP) were stable up to 140 min and intracellular pH (pHi) was maintained and decreased oxygen partial pressure (P(O ( 2) )) from 30 to 15 and 600 Pa produced a significant fall in the intensity of NTP resonance, balanced by an increase in the Pi signal. Treatment of RBC with 0. 5 M isoproterenol during hypoxia exposure did not affect the pattern of changes in NTP or pHi induced by hypoxia and the effect was manifest only on Pi levels.

Atlantic salmon Salmo salar microsatellite markers from a large database were analysed and selected with technical, economic and genetic criteria to provide an optimized set of polymorphic DNA markers for the analysis of the genetic diversity and the structure of anadromous Atlantic salmon populations. A set of 37 microsatellite markers was identified that are easy to use and provide a high level of differentiation power.

A flexible panel consisting of 38 informative microsatellite markers for Salmo trutta is described. These markers were selected from a pool of over 150 candidate loci that can be readily amplified in four multiplex PCR groups but other permutations are also possible. The basic properties of each markers were assessed in six population samples from both the Burrishoole catchment, in the west of Ireland, and Lough Neagh, in Northern Ireland. A method to assess the relative utility of individual markers for the detection of population genetic structuring is also described. Given its flexibility, technical reliability and high degree of informativeness, the use of this panel of markers is advocated as a standard for S. trutta genetic studies.

Fish assemblage structure, rarefied species richness, species diversity and evenness of assemblages upstream of a reservoir in Oklahoma, U.S.A., were compared pre and post-impoundment as well as in contemporary collections from streams above and below the reservoir. There were significant shifts in assemblage structure between historical and contemporary collections above the reservoir but not between contemporary assemblages above and below the impoundment. Indicator species analysis revealed that the sand shiner Notropis stramineus and fathead minnow Pimephales promelas have declined, whereas largemouth bass Micropterus salmoides and western mosquitofish Gambusia affinis have increased in relative abundance in assemblages upstream of the impoundment. Species richness was lower in contemporary assemblages compared with historical assemblages. Furthermore, contemporary assemblages below the dam had lower species richness, diversity and evenness compared with contemporary collections above the dam. These results highlight the spatial and temporal extent of reservoirs altering fish assemblages upstream of impoundments.

Estimated pike Esox lucius recruitment varied by a factor of 16 for females from 1944 to 1991 and by a factor of 27 for males from 1943 to 1990 in Windermere, a temperate, mesotrophic U.K. lake. No significant stock-recruitment relationships were found, but analysis with general additive models (GAMs) revealed that early autumnal water temperature, strength and direction of the North Atlantic Oscillation displacement (corresponding to different climatic conditions in winter) and zooplankton abundance but above all, late summer water temperature were important explanatory variables over the entire time series. Female recruitment was also influenced by young-of-the-year winter temperature. There was no evidence that perch Perca fluviatilis year-class strength, lake level or the summer position of the Gulf Stream influenced recruitment. The fitted models explained up to c. 65% of the overall observed variation between years.

Sixty novel simple sequence repeat (SSR) markers were developed from expressed sequence tags (EST) of half-smooth tongue sole Cynoglossus semilaevis exploited in the laboratory. The number of alleles, observed and expected heterozygosity per locus ranged from two to 16, from 0·0833 to 1·0000 and from 0·0816 to 0·913, respectively. Of these SSRs, 20 had significant homology to known genes by BLASTx (basic local alignment search tool x) search. For cross-species amplification, there are 53 positive amplifications in Japanese flounder Paralichthys olivaceus with 12 polymorphic loci and 51 positive amplifications in Senegalese sole Solea senegalensis with 11 polymorphic loci. These new EST-SSR markers will be useful for genetic studies and genome mapping of C. semilaevis and its closely related fishes.

A survey of 65 populations of chum salmon Oncorhynchus keta across the species range revealed homozygote excess (947 homozygotes in 2954 fish) at a polymerase chain reaction (PCR)-based simple sequence repeat (SSR) locus oke3 with multiple alleles, whereas re-designed PCR primers indicated that 328 of these homozygotes were actually heterozygotes. Statistically significant high positive values of inbreeding coefficients, f, in multiple populations appeared to be a reliable predictor of null alleles. Based on these data, three methods were checked for their ability to estimate null-allele frequencies. © 2014 The Fisheries Society of the British Isles.

In this study, the complementary (c)DNA encoding heat-shock protein 70 (Hsp70) of orange-spotted grouper Epinephelus coioides (OsgHsp70) was cloned. OsgHsp70 was 2206 bp and encoded 652 amino acids with predicted molecular mass of 70·89 kDa and theoretical isoelectric point of 5·48. Three Hsp70 family signatures, bipartite nuclear localization signal sequence (NLS) and cytoplasmic characteristic motif (EEVD) were observed in the OsgHsp70, which shared high similarity in amino-acid sequences with the Hsp70 gene of other vertebrates. The results indicated that the OsgHsp70 is a member of the heat-shock protein 70 family. The Hsp70 messenger (m)RNA expressions were quantified by real-time PCR following heat shock, bacterial infection and immunization with formalin-killed Vibrio alginolyticus, a kind of bacterial pathogen that causes septicaemia. Hsp70 mRNA expression in gill, kidney, spleen, thymus gland, muscle and total-blood samples increased at first and then decreased gradually following heat shock. A similar time-dependent pattern was observed following V. alginolyticus pathogen challenge, in which Hsp70 mRNA expression peaked at 24 h after live bacterial infection and 3 days after dead bacterial vaccination. The results indicated that the Hsp70 gene was inducible and involved in the fish immune response.

A full-length MyD88 cDNA (CiMyD88) was cloned and characterized from grass carp Ctenopharyngodon idella. CiMyD88 was found to be broadly expressed and was up-regulated by grass carp reovirus (GCRV) and CiMyD88 transcripts in vitro were rapidly elevated in C. idella kidney (CIK) cells after challenge with poly(I:C). These results suggest that CiMyD88 may be involved in the antiviral immune defence in C. idella.

The full-length cDNA of heat shock protein 90 (HSP90) of humphead snapper Lutjanus sanguineus, designated as rsHSP90, was cloned by rapid amplification of complementary (c)DNA ends (RACE) techniques with the primers designed from the known expressed sequence tag (EST) sequence identified from the subtracted cDNA library of the head kidney of L. sanguineus. Sequence analysis showed that the full-length cDNA of rsHSP90 was 2745 bp, containing a 5' terminal untranslated region (UTR) of 99 bp, a 3' terminal UTR of 471 bp and an open reading frame (ORF) of 2175 bp encoding a polypeptide of 725 amino acids. On the basis of the deduced amino acid sequence, the theoretical molecular mass of rsHSP90 was calculated to be 83·18 kDa with an isoelectric point of 4·79. Moreover, five classical HSP90 family signatures were found in the amino acids sequence of rsHSP90 by PredictProtein. Basic local-alignment search-tool (BLAST) analysis revealed that the amino acids sequence of rsHSP90 had the highest similarity of 97% when compared with other HSP90s. Fluorescent real-time quantitative reverse-transcription (RT)-PCR was used to examine the expression pattern of rsHSP90 in eight kinds of tissues and organs of L. sanguineus challenged with Vibrio harveyi. There was a clear time-dependent expression pattern of rsHSP90 in head kidney, spleen and thymus after bacterial challenge and the expression of messenger (m)RNA reached the maximum level at the time points of 9, 15 and 24 h, respectively. The up-regulated mRNA expression of rsHSP90 in L. sanguineus after bacterial challenge indicated that rsHSP90 was inducible and might be involved in immune response.

Abant trout Salmo trutta abanticus were grown for 154 days at three salinities of 0, 9 or 18 to assess mass gain, condition factor, food conversion ratio and food consumption. Appetite was significantly higher at 0 than at salinities of 9 or 18. Salinities >18 may have detrimental effects on biomass production. The mean ± S.E. of the condition factor at the three salinities were 1.29 ± 0.20 at 0, 1.22 ± 0.22 at 9 and 1.10 ± 0.20 at 18. Fish reared at a salinity of 0 showed higher mass gain, food conversion ratio and food consumption.

This study provides data on the genetic structuring of the pipefish Syngnathus abaster in the western Mediterranean and Adriatic Seas. A total of 109 specimens were collected in brackish-water biotopes. The control region and three other regions of the mitochondrial genome were analysed. The most relevant result was the high genetic structuring found by Bayesian inference (BI), maximum likelihood (ML) and network analyses, which were consistent in showing three well-separated clusters of S. abaster populations. Furthermore, BI and ML did not support the monophyly of the taxon S. abaster. These results suggest the occurrence of a species complex in the study area, whose differentiation may have occurred since the Pleistocene. The results also show a very high genetic variability at the inter-population level, with no shared haplotypes among sites. Evolutionary forces due to the fragmented nature of the brackish-water habitats may account for the high genetic divergence found among the groups and populations. Finally, although dispersal by rafting over long distances may occasionally occur, this study suggests linear stepping-stone model of colonization to be most likely. The complexity of the results obtained suggests that further studies are needed to elucidate the phylogeny of S. abaster.

A method is described to assess the reproductive status of male Hippocampus abdominalis on the basis of behavioural traits. The non-invasive nature of this technique minimizes handling stress and reduces sampling requirements for experimental work. It represents a useful tool to assist researchers in sample collection for studies of reproduction and development in viviparous syngnathids, which are emerging as important model species.

The influence of relatedness on male-male aggression was tested in three-spined sticklebacks Gasterosteus aculeatus. The intensity of aggression against brothers and non-kin males did not differ significantly, indicating that kin recognition plays at most a minor role in aggressive interactions between male G. aculeatus.

Residues of maternal nuclear DNA in the form of chromosome fragments were observed in the healthy and morphologically normal androgenetic rainbow trout Oncorhynchus mykiss. A hypothetical model for formation of chromosome re-arrangements caused by the incomplete maternal nuclear DNA inactivation in the androgenetic rainbow trout was proposed in the present paper.

The auditory abilities of the round goby Neogobius melanostomus were quantified using auditory evoked potential recordings, using tone bursts and conspecific call stimuli. Fish were tested over a range of sizes to assess effects of growth on hearing ability. Tests were also run with and without background noise to assess the potential effects of masking in a natural setting. Neogobius melanostomus detected tone bursts from 100 to 600 Hz with no clear best frequency in the pressure domain but were most sensitive to 100 Hz tone stimuli when examined in terms of particle acceleration. Responses to a portion of the N. melanostomus call occurred at a significantly lower threshold than responses to pure tone stimulation. There was no effect of size on N. melanostomus hearing ability, perhaps due to growth of the otolith keeping pace with growth of the auditory epithelium. Neogobius melanostomus were masked by both ambient noise and white noise, but not until sound pressure levels were relatively high, having a 5-10 dB threshold shift at noise levels of 150 dB re 1 µPa and higher but not at lower noise levels.