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The Chinese mitten crab (Eriocheir sinensis), an economically valuable crustacean that is popular for its flavor, exhibits catadromous spawning migration. Overfishing and environmental pollution have inflicted serious damage on wild E. sinensis populations, and the Chinese government has banned the commercial fishing of this species in the Yangtze River. Studies have examined the sexual dimorphism in the body size and morphology of crabs, but there are few reports on the molecular regulatory mechanisms that occur during the reproduction of E. sinensis. In this study, we performed the first comparative transcriptome analyses of the cerebral ganglion and hepatopancreas of E. sinensis during reproduction. The results indicate that E. sinensis has significant sexual dimorphism in signal transduction, metabolism, substance transportation, and cellular protection. This study aims to provide information that can be used as a basis for further research on the molecular mechanisms that underlie sexual dimorphism in E. sinensis during reproduction. Furthermore, the results can be used to support the development of the E. sinensis breeding industry and the restoration of wild E. sinensis. • This study revealed sexual dimorphism in signal transduction and metabolism in Eriocheir sinensis under reproduction. • This study provided key pathways and DEGs in regulation of sexual dimorphism in Eriocheir sinensis under reproduction. • This study provided basic information for further research on regulation of reproduction of Eriocheir sinensis. [ABSTRACT FROM AUTHOR]

Coastal zones are productive areas of marine ecosystems which are also hotspots of anthropogenic activities causing habitat degradation. In the southwest Baltic Sea, eutrophication is thought to have caused the massive reduction in submerged macrophytes observed in recent decades. Here, we surveyed the submarine vegetation and examined locations of spawning of herring ( Clupea harengus ) in the Greifswalder Bodden, one of the most important reproductive habitats of the Western Baltic Spring Spawner herring stock (WBSS). This stock deposits eggs onto submerged vegetation and changes in macrophyte coverage are expected to influence the availability of reproductive habitat. Aerial, underwater video tows and SCUBA surveys conducted in spring 2009 revealed that only ∼7% of the lagoon was vegetated. Herring eggs were observed on 12 of 32 SCUBA transects, at depths between 0.2 and 5 m and were attached to a variety of spermatophyte and algae species but not to stones or mussels. A classification tree model indicated that spawning sites were strongly associated with the vegetation cover within a 100- and 500-m radius, implying that herring schools preferentially spawn on dense and large underwater meadows. Only ∼5% of the lagoon now falls into this vegetation category. Despite 20 years of efforts to reduce eutrophication, no increase in macroalgae and spermatophyte vegetation towards the historical level of 90% coverage in the area is apparent. [ABSTRACT FROM AUTHOR]

Investigations on sexual reproduction of jellyfish are essential to understanding mechanisms and patterns of outbreaks formation. Pelagia noctiluca (Forskål, 1775) (Scyphozoa) is known as the predominant jellyfish species with direct development in Western and Central Mediterranean Sea. In this paper we used integrated morphometric, histological, and biochemical approaches to investigate the annual reproductive biology of P. noctiluca from the Strait of Messina (South Thyrrenian Sea), a key proliferation area for this species due to favourable temperatures and high productivity. From November 2011 to September 2012, P. noctiluca sexual reproduction occurred throughout the year, with two seasonal peaks (autumn, spring) of spawning and embryonic development. Gonads of female P. noctiluca were characterized by a large amount of mature eggs of small size (diameter < 200 μm) during high food availability, whereas fewer, larger eggs (diameter > 200 μm) were detected during low availability of prey. Two morphometric indexes were applied: the Gonad-Somatic Index (GSI, gonadal/somatic tissue dry weight ratio) and Fecundity Index (FI, n° eggs mm −2 * gonadal dry weight). The FI showed longer spawning periods than the GSI, providing a better causal-mechanistic explanation for the year-round occurrence of P. noctiluca in the Strait of Messina. Protein contents of the gonads changed seasonally, with the highest concentrations during the pre-spawning periods. We suggest that investigations on jellyfish sexual reproduction can provide biological information relevant for understanding mechanisms of jellyfish blooms as well as for the management of coastal zones affected by outbreaks of gelatinous species. [ABSTRACT FROM AUTHOR]

Fish species classified as demersal substrate spawners attach their eggs on macrophytes or hard substratum. As a result of consecutive events of egg deposition and successive waves of spawning adults, the formation of multiple egg layers on spawning substrate was already observed for demersal spawning fish species, such as the Atlantic herring (Clupea harengus, Linnaeus 1758). However, empirical information on the effects of egg concentration on herring egg survival are scarce. Hypothesizing that high egg concentration has a negative effect on herring egg survival, we experimentally investigated egg mortalities in artificially spawned egg clusters of 1–10 vertical layers. Moreover, potential effects of egg packing density (loose and dense) in one single layer set-up were investigated. We found decreasing herring egg survival in clusters of increasing egg layers. Although egg mortality was lowest in surface layers, a layerwise analysis did not reveal a stepwise increase of egg mortality from outermost to innermost eggs. Furthermore, egg mortality in surface layers significantly increased with increasing egg layer number underneath, indicating that the extent of layering is affecting egg mortality also in surface layers. This implies a reproductive disadvantage not only for eggs being spawned in the beginning of consecutive spawning events but also for latest spawned eggs. However, different egg packing densities in single layer spawn did neither influence the fertilization success nor mortality of herring eggs. Increased thickness of egg layers may result from the ongoing reduction and fragmentation of coastal vegetation due to eutrophication and coastal modifications. Our results indicate that such multiple egg layer deposition has the potential to negatively impact hatching success in demersal spawning fish species, such as the herring. [Display omitted] • We investigated density-dependent herring egg mortality via laboratory experiments. • The total egg mortality increases with the number of layers within batches of spawn. • Surface layer mortality is lowest but increases with numbers of egg layers underneath. • Monolayers result in best egg survival but require more area of spawning substrate. • Habitat loss related increase in spawn concentrations pose a threat for egg survival. [ABSTRACT FROM AUTHOR]

Most publications devoted to the cryopreservation of mollusc sperm have focused on the definition of technical protocols, avoiding the description of sperm quality after thawing. The present study investigated the effects of cryopreservation on sperm quality in the great scallop. Wild scallop were fished during the natural spawning period and conditioned in the hatchery before use. Sperm samples were obtained after intragonadal injection of serotonin and cryopreserved using a previously published protocol. Sperm quality was assessed using a panel of four parameters: sperm motility characteristics, using a computer assisted sperm analysis plugin with Image J, intracellular ATP content using an ATP-Lite kit, sperm integrity, using flow cytometry and sperm morphology, using transmission electron microscopy. For each parameter, fresh (control) and thawed spermatozoa were compared. A significant decrease of both the percentage of motile spermatozoa (reduction: 75%) and sperm swimming speed (86%) were observed for thawed sperm compared with fresh sperm. The percentage of living spermatozoa, as assessed using flow cytometry, was significantly lower for thawed sperm (72.4 ± 2.5%) compared with fresh sperm (86.4 ± 1.1). However, no significant difference of intracellular sperm ATP content was observed between fresh and thawed sperm. Post thawing, while some spermatozoa showed little or no morphological differences compared with fresh sperm, others had undergone drastic changes, including swelling of the plasma membrane, structural alterations of the chromatin and damage to mitochondria. In conclusion, the descriptive parameters studied in the present work showed that the quality of thawed great scallop sperm was lower than that of fresh cells but was still sufficient for use in aquaculture programs and sperm cryobanking for this species. [ABSTRACT FROM AUTHOR]

Abstract: In the present study, we developed and validated real-time quantitative RT-PCR assays for a suite of genes involved in the brain-pituitary gonadal axis in fish including kisspeptin genes and its receptor (Kiss1, kiss2, kissr4) and gonadotropin-releasing hormone genes (sbGnRH, sGnRH, cGnRHII) in the brain, and gonadotropin genes (fshβ and lhβ) in the pituitary. Sex steroid profiles (T and 11-KT) and gonadal development were also studied over a full annual reproductive cycle in adult male sea bass. The cDNA partial sequence of sea bass kissr4 encoding 185 amino acids showed a high degree of conservation with other fish kissr4 subtype. Results clearly showed a seasonal profile for Kiss1, kiss2 and kissr4 mRNAs. Kissr4, fshβ and lhβ levels increased gradually and peaked during spermatogenesis (January) while Kiss1, kiss2, cGnRH-II as well as steroids showed peaks during early spawning (March). No significant seasonal changes were observed for sbGnRH and sGnRH expression. These results support the possible involvement of the kiss genes and their receptor (kissr4) in the seasonal control sea bass reproduction. However, a lack of correlation between kiss genes and sbGnRH expression and the mismatch between kisspeptin and the onset of gonadotropin surge contrast with previous findings. [Copyright &y& Elsevier]

Cryopreservation can be a powerful tool in shellfish aquaculture for selective breeding programmes and hatchery production. Most species that are commercially farmed are broadcast spawners, releasing sperm and eggs into the environment where they fertilize and develop. However, some species are brooders (e.g. flat oysters (Ostrea chilensis)), which retain and rear embryos until they reach later developmental stages. Sperm cryopreservation has been successfully achieved for several farmed species including the Pacific oyster (Crassostrea gigas), greenshell™ mussel (Perna canaliculus) and abalone (Haliotis iris). In general, dimethyl sulphoxide and trehalose are the preferred cryoprotectants (CPAs) and simple methods to control cooling rate (e.g. floating rack on liquid nitrogen; methanol/dry ice bath) have worked well for our main species of interest (Pacific oyster, greenshell™ mussel and abalone). Interestingly, sperm perform better in CPAs that are prepared in Milli-Q water as opposed to seawater, indicating that a lower salt concentration during freezing is beneficial. Recent studies with flat oysters and geoducks (Panopea zelandica) have also resulted in successful sperm cryopreservation. Shellfish oocyte and larvae cryopreservation has proven more challenging, however, successful cryopreservation of Pacific oyster oocytes has been achieved. Pacific oyster oocytes are able to be fertilized and develop to normal spat post-thawing. However, for other species (e.g. greenshell™ mussel) and for larvae, cryo-injuries are not always immediately apparent post-thawing but manifest during development and are often lethal. greenshell™ mussel oocytes are irretrievably damaged when they reach -6 to -8°C and attempts to overcome this sub-zero chilling injury by vitrification have been unsuccessful. Towards understanding these injuries and their effects, we investigated the effect of cryopreservation on the meiotic spindle of greenshell™ mussel oocytes. Results show that the meiotic spindle undergoes major changes during both CPA addition and cooling. However, the spindle is able to re-form normally in most instances given sufficient time post thawing. For a proportion of oocytes, cryopreservation appears to trigger oocyte activation. The effect of cryopreservation on oxidative stress in greenshell™ mussel oocytes has also been investigated. Although, there are changes in oxidative stress parameters as a result of cryopreservation and strong correlations between fertilization, development and oxidative stress parameters (lipid hydroperoxides, superoxide dismutase, protein carbonyls, total glutathione and glutathione state), preliminary attempts to mitigate oxidative stress using antioxidants have so far failed to enable normal development following cooling, suggesting that oxidative stress may be a secondary rather than primary stress event. Research on shellfish larval cryopreservation utilizing SEM and confocal microscopy has also been carried out for greenshell™ mussel. Later larval stages (D-larvae) appear more resilient to cryopreservation than trochophore larvae, but abnormalities are still apparent (e.g.shell abnormalities and delayed organogenesis). Future studies are aimed toward elucidating mechanisms of injury using new tools such as proteomics and then targeting susceptible cellular processes for protection during cryopreservation. Source of funding: This research was funded by the New Zealand Ministry of Business Innovation and Employment (CAWX0802). Conflict of interest: None declared. serean.adams@cawthron.org.nz [ABSTRACT FROM AUTHOR]