A molecular tool for parentage analysis in the Mediterranean mussel (Mytilus galloprovincialis)

Estimation of selection response and genealogical tracing in family mixtures require an appropriate tool for parentage analysis. In this study, we tested 19 marker loci for parentage analysis allocation in Mediterranean mussel (Mytilus galloprovincialis). To this aim, we reared families in tanks isolated from wild mussel seed, analysed them using the 19 marker loci and characterized their performances based on Mendelian rules. Probabilities of exclusion of a false parent were estimated for different groups of loci and contrasted to the real paternity assignment. Based on this, we chose nine microsatellites with the highest exclusion probabilities and a real paternity assignment of 99.6%. Next, we analysed 600 individuals reared as in the usual production process, where contamination from wild seed is likely. We obtained a real assignment of 94.7% and were able to identify individuals from the wild as the most likely hypothesis to explain the observed incompatibilities with candidate parents. This information was used to evaluate parental contribution in offspring obtained from gamete mixtures of several parents, which bestowed results of interest for future breeding programs of Mediterranean mussel.     

DNA fragmentation in blue mussel (Mytilus edulis) sperm: aquaculture and fisheries implications

The objective of this study was to assess sperm DNA longevity in blue mussels (Mytilus edulis) using a dynamic assessment of sperm DNA fragmentation (SDF) after sperm activation. Mature blue mussels (n = 57) in Vigo (Galicia, Spain) were obtained, specifically rope farmed blue mussels (n = 38) and wild blue mussels (n = 19). After the sperm collection, a subsample was assessed for SDF (0 h), while the rest of the sample was incubated for 6, 9, 12, 24 and 48 h at 15°C, assessing each time point using the Sperm‐Halomax kit (Halotech DNA, Madrid, Spain). The Kaplan–Meier estimator, log‐rank (Mantel–Cox) test and Mann–Whitney U‐test were used for statistical analyses (spss v. 16.0), α = 0.05. The rate of SDF (r‐SDF) between rope farmed and wild blue mussels over 0–6 h incubation was not significantly different (P = 0.278), but was for 6–24 h (P = 0.004). Differences in r‐SDF were observed when comparing the means between the two groups (P < 0.0001). Individual differences in r‐SDF existed among the rope farmed (P < 0.0001) and wild blue mussels (P < 0.0001). Wild blue mussels presented a higher DNA longevity than the farmed blue mussels. Selection of blue mussel males with a low level of sperm DNA damage and greater sperm DNA longevity may result in better fertilization and seed production.     

Cryopreservation of strip spawned sperm using non‐programmable freezing technique in the blue mussel Mytilus galloprovincialis

In this study, a non‐programmable freezing technique has been developed with the strip spawned blue mussel (Mytilus galloprovincialis) sperm. The key parameters optimized including (1) cryoprotectant agents (CPAs); (2) cooling temperature; (3) thawing temperature; (4) sugar and amino acid supplementation and (5) sperm to egg ratio. The fertilization rate and/or integrity of plasma membrane and acrosome were used as sperm quality assessment indicators. The highest post‐thaw sperm fertilization rate of 95% was achieved, when sperm were cryopreservated in 8% dimethyl sulfoxide at 7.8 cm above the liquid nitrogen surface and thawed in a 60°C seawater bath. The addition of glucose, sucrose or trehalose in dimethyl sulfoxide did not, whereas 0.8% glycine did significantly improve the post‐thaw sperm fertilization rates. The fluorescent evaluation has demonstrated that the addition of glycine significantly improved the post‐thaw sperm acrosome integrity, revealing a positive role of glycine in the improvement of post‐thaw sperm quality in blue mussels.     

Effects of environmental factors and food availability in Northern Aegean sea on the cultivation of Mediterranean Mussels (Mytilus galloprovincialis)

This study used two different collectors made of polypropylene (PP) rope and polyethylene (PE) fishing net to determine Mediterranean mussel's settlement, growth, condition index (CI), meat yield (MY) and shell thickness index (STI) during the period spent between June 2017 and November 2018. With this regard, chlorophyll a, water temperature, salinity, pH and particulate matter were defined through water samples that were collected on monthly basis triplicate. The mean chlorophyll a level was recorded as 0.32 ± 0.31 μg/L, while water temperature was 19.73 ± 5.14°C, salinity was 35 ± 2.070‰, pH was 8.12 ± 0.04, and TPM was 14.91 ± 10.48 mg/L. As a result of the cultivation period of 8 months spent following the first intense grip, the length of the mussels is measured as 31.79 ± 6.20 mm. Based on the Pearson correlation analysis results, the most important environmental parameter affecting Mediterranean mussels growth in length on PP rope collectors is the temperature (p < .05). The STI and CI were determined to be related with environmental temperature parameters, while no determinations could be obtained concerning the correlation of MY with environmental parameters. Assessment of Mediterranean mussels’ growth rate in PP rope collectors proved significant differences (p < .05) between March and November 2018. As no Mediterranean mussels are observed on PE fishing nets during the samplings performed throughout the study period, it is determined that PE fishing nets are not suitable collector types for Mediterranean mussel cultivation.     

Quantification of waste feed and fish faeces in sediments beneath yellowtail pens and possibility to reduce waste loading by co‐culturing with sea cucumbers: an isotopic study

To evaluate environmental impacts of yellowtail culture and to examine the efficiency of the sea cucumber Apostichopus japonicus in reduction of waste loading, surface sediments were sampled and A. japonicus were cultured at a yellowtail farm in Owase Bay, Japan. Waste feed‐ and faeces‐derived organic matter (WOM, FOM) in sedimentary organic matter (SOM) was estimated based on the δ¹³C and δ¹⁵N of the fish feed (?21.7‰, 9.2‰), yellowtail faeces (?20.6‰, 6.2‰) and SOM reference (?24.4‰, 4.4‰). Small WOM (0–22% in SOM) but substantial FOM (28–61%) loadings in the fish farm area and high sulphide concentrations in the sediments (1.0 mg g^?1) suggest that reduction in the fish stocks or mitigation of the faeces should be considered. A. japonicus juveniles were cultured in three cages deployed below a pen and growth was assessed after three different periods (62, 107, 181 days). A. japonicus grew well during the first 107 days (daily specific growth rate, 3.7%) and their survivorship was high (80–90%). Growth ceased after 107 days, probably due to fouling on the cages. The δ¹³C and δ¹⁵N of their hypothetical diet (–19.7‰, 5.5‰) were close to the FOM values, suggesting assimilation of FOM.     

The interaction of infectious salmon anaemia virus (ISAV) with the blue mussel,Mytilus edulis

Integrated multi‐trophic aquaculture (IMTA) is an alternative approach to mono‐culture aquaculture that reduces environmental impacts of commercial aquaculture systems by combining the cultivation of fed species with extractive species. Shellfish play a critical role in IMTA systems by filter‐feeding particulate‐bound organic nutrients. They may also increase or decrease disease risk on farms by serving as reservoirs or barriers for important finfish pathogens such as infectious salmon anaemia virus (ISAV). This study aimed to optimize culture and molecular assays in shellfish tissues and to determine the fate of ISAV in mussels, Mytilus edulis. To determine detection limits, qRT‐PCR and culture assays in both CHSE‐ and ASK cells were optimized in ISAV‐inoculated mussel tissue homogenates. Both qRT‐PCR and culture assays performed in ASK cells had comparable detection limits of 10^2.8 TCID₅₀ mL^?1. The ISAV RNA genome was consistently detected in digestive gland tissue of ISAV‐exposed mussels. Viable ISAV was not detected in mussel tissues by culture analysis in CHSE‐ and ASK cells. The fact that qRT‐PCR analysis resulted in positive cycle threshold (CT) values that corresponded to the detectable range of ISAV in ASK culture assays suggests that little to no viable ISAV particles are present in the mussel tissues.     

亚热带网箱区底播仿刺参养殖周期内生理生态学特征

在现场条件下研究了亚热带网箱区底播养殖仿刺参的生理生态学指标随时间的变化。结果显示,网箱区底播的仿刺参耗氧率变化范围为0.012~0.016 mg O2/(g·h),各时期数据间无显著差异,表明在实验期间仿刺参维持基本代谢的能量消耗随时间变化不大;对照区5月仿刺参的耗氧率显著高于2、3、6月,表明对照区仿刺参夏眠初期能量消耗较大。2、3月网箱区仿刺参的耗氧率显著高于同期对照区,表明冬季网箱区底播仿刺参代谢更为活跃。网箱区底播仿刺参排氨率随着时间不断下降,2月排氨率[(0.061±0.009)μmol/(g·h)]显著高于5月[(0.045±0.011)μmol/(g·h)])和6月[(0.035±0.007)μmol/(g·h)],这是因为冬、春季节水温更适宜仿刺参生长,该阶段仿刺参摄食活跃,可以有效地利用网箱区高质量的食物。5月以后网箱区仿刺参进入夏眠状态,肠道逐渐萎缩,其排氨率也随之降低。对照区仿刺参排氨率始终维持在较低的水平,表明其营养状态不佳。各时期网箱区和对照区的O/N比值均为10~30,表明仿刺参新陈代谢底物包含蛋白质和碳水化合物或者脂质。其中,2—5月网箱区刺参O/N比值相对于对照区较低,表明该阶段网箱区仿刺参新陈代谢底物中蛋白质所占比例较多,这与该时期网箱区较高的食物质量相对应。     

Development and validation of a range of endogenous controls to support the implementation of practical Taqman real-time PCR-based surveillance for fish diseases within aquaculture

The use of Taqman real-time PCR-based technology has recently become more frequent in the detection of pathogens in the aquaculture industry. This interest has necessitated the development of robust and reliable pathogen-detection assays. The development of a range of endogenous control assays to be run alongside these diagnostic assays works to further increase confidence in the latter. This study describes the design of a range of endogenous control assays based on the elongation factor 1-α (EF1-α) gene specific to a range of fish species including Atlantic salmon, Salmo salar; rainbow trout, Oncorhynchus mykiss; brown trout, Salmo trutta; cod, Gadus morhua; haddock, Melanogrammus aeglefinus; saithe, Pollachius virens; whiting, Merlangius merlangus; Norway pout, Trisopterus esmarkii; carp (family Cyprinidae), roach, Rutilus rutilus; European eel, Anguilla anguilla; and herring, Clupea harengus, as well as a number of fish cell lines. Evidence is provided of the validation of these assays for specific species, a range of tissue types and cell lines as well as an example of the potential uses of these assays.     

The effect of density in larval rearing of the pullet carpet shell Venerupis corrugata (Gmelin, 1791) in a recirculating aquaculture system

The pullet carpet shell Venerupis corrugata is an economically valuable species in several European countries, however, nowadays stocks are under high fishing pressure. Hatchery production of juveniles for release is a major contributor to strengthen the stock and consequently improve the sustainability of the natural stocks. This study aimed to determine the feasibility of rearing V. corrugata larvae with different larval densities (10, 40 and 200 larvae per mL) in a recirculating aquaculture system (RAS), compared with the traditional larval rearing methodology (Batch). The mean survival, growth and metamorphic rate of V. corrugata larvae in RAS was higher (11.1%; 71.3 μm; 21.6% respectively) than in the Batch system, in all tested densities. The larval growth was not affected by the initial density until 40 larvae per mL, however, 200 larvae per mL decreased the larval growth in length nearly 54 μm. The larval rearing time was shortened in 2 days in the RAS system. The physical, chemical and microbiologic parameters suggested that the tested densities were not excessive to disturb the biofilter stability of RAS. The V. corrugata larval rearing performed at high larval stocking densities in RAS system present a reduction in the operating costs to produce this species.     

Impact of sublethal di‐n‐butyl phthalate on the aquaculture fish species Nile tilapia (Oreochromis niloticus): histopathology and oxidative stress assessment

Phthalates such as di‐n‐butyl phthalate (DBP) and their esters are widely used plasticizers, their ubiquitous presence in daily life, inevitably leads to their restricted use due to important environmental pollution and health impacts and endocrine disruption potential. The aim of this study was to examine the effects of a sublethal concentration of 10 mg L^?1 DBP on haematocrit (HCT) values, gills and liver histology, malondialdehyde (MDA, 2‐thiobarbituric acid‐TBA reactivity) and reduced glutathione (GSH) levels in gills and liver tissue as oxidative stress biomarkers in the aquaculture fish species Nile tilapia (Oreochromis niloticus) after 24 (DBP‐24) and 96 (DBP‐96) h exposure. No differences were found between per cent HCT values in the 24 h exposure groups (P > 0.05). Response of antioxidant defence systems in liver and gill tissues of the fish were dependent on exposure duration and changed to a higher extent during 96 h. MDA levels in liver tissue increased in DBP treated fish in comparison to the control fish. However, the differences between the exposure and control groups were not significant (P > 0.05). A statistically significant decrease (P > 0.05) was recorded in gill MDA levels in the DBP‐96 group when compared to the control and DBP‐24 groups. The liver GSH levels were unchanged in the DBP treated fish. However, GSH levels were significantly lower in the gill tissue of the DBP‐96 group. Exposure to DBP caused several degenerative changes in the histology of gill and liver tissue. Gills displayed hyperaemia, epithelial lifting, oedema, talengiectasia, epithelial hyperplasia and fusion of secondary lamellae, whereas in liver several circulatory anomalies (hyperaemia, blood congestion and sinosoid dilatation) and vacuolization of hepatocytes were observed. Histopathological results demonstrated that the gills were more affected than the liver perhaps due to their direct contact with DBP.     

Effects of black soldier fly (Hermetia illucens) larvae meal protein as a fishmeal replacement on the growth and immune index of yellow catfish (Pelteobagrus fulvidraco)

A 65 days feeding trial was conducted to evaluate the potential of black soldier fly larvae (BSFL) as a fish meal (FM) protein alternative in the diet of yellow catfish. Fish meal protein was replaced with BSFL meal protein at 0% (control), 13%, 25%, 37%, 48%, 68%, 85% and 100%. Compared with the control group, increased growth performances (e.g. weight gain rate increased by 21.7%) and immune indexes (e.g. serum lysozyme activity increased by 6.8%) of yellow catfish fed with diets in which a maximum of 48% FM protein was replaced by BSFL meal protein was determined. The diet in which 25% FM protein was replaced by BSFL meal protein resulted in the greatest growth performances (e.g. weight gain rate increased by 29.1%) and immune indexes (e.g. serum lysozyme activity increased by 31.9%) as well as the lowest feed conversion ratio (0.9) among all diets tested. No significant differences in survival rate, body indexes or composition were determined among all treatments. Therefore, BSFL meal protein has the potential to partially replace FM protein in the diet of yellow catfish, and it may also enhance the immunocompetence of the fish.     

The use of bromelain as a feed additive in fish diets: Growth performance,intestinal morphology,digestive enzyme and immune response of juvenile Sterlet (Acipenser ruthenus)

Bromelain is a proteolytic enzyme extracted from Ananas comosus and has great potential to affect several physiological functions. The current study examined the effect of bromelain added to commercial feed at concentrations of 0 g (control), 10 g (B1) and 20 g (B2) per kg diet on growth, feed utilization, intestinal morphology, digestive enzyme and immune response of juvenile Sterlet (Acipenser ruthenus) for 56 days. At the end of this experiment, the highest final body weight was detected in fish fed diet of B2 compared to control. The protein content of whole fish was higher in fish fed diet of B1 and B2, but the content of calcium, iron, copper and zinc was noted lower in fillets of fish fed diet of B1 and B2. Supplementation with bromelain significantly increased the height of mucosal folds, enterocytes and the supranuclear zone of the epithelial cells compared to control diet. The enzymatic activity of lipase and pepsin was significantly (p < 0.05) higher in fish fed diet of B1 and B2. The highest activity of lysozyme, total protein level and total immunoglobulin and the proliferative activity of T and B cells were detected in fish fed diet of B2 compared to control, where no significant (p > 0.05) difference was found in ceruloplasmin, metabolic activity of spleen macrophages and potential killing activity of spleen phagocytes between different treatments.     

The role of sand lances (Ammodytes sp.) in the Northwest Atlantic Ecosystem: A synthesis of current knowledge with implications for conservation and management

The American sand lance (Ammodytes americanus, Ammodytidae) and the Northern sand lance (A. dubius, Ammodytidae) are small forage fishes that play an important functional role in the Northwest Atlantic Ocean (NWA). The NWA is a highly dynamic ecosystem currently facing increased risks from climate change, fishing and energy development. We need a better understanding of the biology, population dynamics and ecosystem role of Ammodytes to inform relevant management, climate adaptation and conservation efforts. To meet this need, we synthesized available data on the (a) life history, behaviour and distribution; (b) trophic ecology; (c) threats and vulnerabilities; and (d) ecosystem services role of Ammodytes in the NWA. Overall, 72 regional predators including 45 species of fishes, two squids, 16 seabirds and nine marine mammals were found to consume Ammodytes. Priority research needs identified during this effort include basic information on the patterns and drivers in abundance and distribution of Ammodytes, improved assessments of reproductive biology schedules and investigations of regional sensitivity and resilience to climate change, fishing and habitat disturbance. Food web studies are also needed to evaluate trophic linkages and to assess the consequences of inconsistent zooplankton prey and predator fields on energy flow within the NWA ecosystem. Synthesis results represent the first comprehensive assessment of Ammodytes in the NWA and are intended to inform new research and support regional ecosystem‐based management approaches.     

Tagging: on the use of passive integrated transponder (PIT) tags for the identification of fish

To determine the efficacy of passive integrated transponder (PIT) tags for marking rohu Labeo rohita (Ham.) in the selective breeding programme, a series of experiments has been carried out at the Central Institute of Freshwater Aquaculture (CIFA) under the Indo–Norwegian project of ‘Selective breeding of rohu’. Six groups of rohu fingerlings with weight ranging from 2 g to 20 g were tagged with PIT tags to determine a suitable size range for tagging. Fingerlings weighing 8–15 g were found to be quite suitable for tagging with a PIT tag. Recovery of the PIT tag depends upon the survival of tagged fish under field conditions. Rejection of the PIT tag by rohu was observed to be only 0.05%. Through effective management practice, the survival of tagged fish increased up to 95%, and thus tag loss was minimized.     

Growth response,body composition,plasma metabolites,digestive and antioxidant enzymes activities of Siberian sturgeon (Acipenser baerii,Brandt, 1869) fed different dietary protein and carbohydrate: lipid ratio

Knowing the effect of dietary nutrients on physiological parameters, especially oxidative stress, during fish rearing can be useful to improve fish health and optimize production in aquaculture. In this study, four iso‐energetic (2 × 2 factorial design) diets differing in nutrient composition were submitted to the Siberian sturgeon juveniles, to analyse biochemical and physiological responses. The juveniles were fed for as long as 10 weeks with the following diets: low protein–high carbohydrate (CHO): lipid ratio (LP‐St), high protein–high CHO: Lipid ratio (HP‐St), low protein‐low CHO: Lipid ratio (LP‐L) and high protein‐low CHO: Lipid ratio (HP‐L). It was shown that HP‐L diet significantly induced higher growth performance in the juveniles than LP‐L; however, there was not a significant difference between them with other treatments. HSI was increased by higher dietary CHO: L ratio. The result obviously showed the higher dietary fat led to an increase in sturgeon body fat content. In addition, dietary protein and CHO: lipid ratio significantly affected plasma metabolites (glucose, cortisol, cholesterol and triglyceride) but did not affect IGF‐1 and insulin levels. Based on enzyme activities results, incorporation of high protein (44%) in the diet induced lower amylase activity and carbohydrate digestion in the sturgeon juveniles. Moreover, antioxidant enzyme activities in this species were more influenced by lipid than other nutrients. In conclusion, using appropriate carbohydrates (~30%) in the sturgeon diet helps to spare protein. Furthermore, it is suggested to decrease the amount of lipid in high protein diet, to improve higher sturgeon fillet quality and less oxidative damage in liver.     

Complete repertoire of long‐chain polyunsaturated fatty acids biosynthesis enzymes in a cyprinid,silver barb (Barbonymus gonionotus): Cloning,functional characterization and dietary regulation of Elovl2 and Elovl4

The silver barb (Barbonymus gonionotus) is a valuable cyprinid with good deposition of long‐chain polyunsaturated fatty acids (LC‐PUFA) such as docosahexaenoic acid (DHA). We have previously isolated a fatty acyl desaturase (Fads2) and elongase (Elovl5), which fulfils the production of eicosapentaenoic acid (EPA) and arachidonic acid (ARA) from C18 polyunsaturated fatty acid (PUFA) substrates. The isolation, functional characterization and dietary regulation of two elongases, Elovl2 and Elovl4, are reported here. The Elovl2 displayed the capacity to elongate C20 and C22 PUFA substrates, while the showed low activities towards saturated fatty acids and C22 PUFA substrates. This discovery validates the existence of a complete set of enzymes of LC‐PUFA in silver barb. The elovl2 showed higher expression in liver, in limited dietary LC‐PUFA intake conditions. As for Elovl4, tissue expression showed prominence in eye tissue. We also showed that the expression of both genes was upregulated when fish was fed with diets devoid of LC‐PUFA. Fatty acid composition analysis indicates the utilization of C18 polyunsaturated fatty acids for LC‐PUFA biosynthesis.