Piscine reovirus in wild and farmed salmonids in British Columbia,Canada: 1974–2013

Piscine reovirus (PRV) was common among wild and farmed salmonids in British Columbia, western Canada, from 1987 to 2013. Salmonid tissues tested for PRV by real‐time rRT‐PCR included sections from archived paraffin blocks from 1974 to 2008 (n = 363) and fresh‐frozen hearts from 2013 (n = 916). The earliest PRV‐positive sample was from a wild‐source steelhead trout, Oncorhynchus mykiss (Walbaum), from 1977. By histopathology (n = 404), no fish had lesions diagnostic for heart and skeletal muscle inflammation (HSMI). In some groups, lymphohistiocytic endocarditis affected a greater proportion of fish with PRV than fish without PRV, but the range of Ct values among affected fish was within the range of Ct values among unaffected fish. Also, fish with the lowest PRV Ct values (18.4–21.7) lacked endocarditis or any other consistent lesion. From 1987 to 1994, the proportion of PRV positives was not significantly different between farmed Atlantic salmon, Salmo salar L. (44% of 48), and wild‐source salmonids (31% of 45). In 2013, the proportion of PRV positives was not significantly different between wild coho salmon, Oncorhynchus kisutch (Walbaum), sampled from British Columbia (5.0% of 60) or the reference region, Alaska, USA (10% of 58).     

Nutritional quality of amino acid in farmed,farm‐aggregated and wild Axillary seabream (Pagellus acarne) with implications to Human Health

This study evaluated the nutritional quality of farmed and wild axillary seabream (Pagellus acarne R., 1827) focusing on amino acid profiles, with regards to possible interactions with wild fish aggregating around the cage facility. Total amino acids (∑AA), essential amino acids (∑EAA), non‐essential amino acids (∑NEAA) and neutral amino acids (∑NAA) in farmed fish were lower than those in the wild individuals (p > .05). Amino acid pattern in cage‐aggregated fish showed a slight decline from the wild populations, but still higher than the farmed fish. Based on the amino acid scores (AAS), lysine and leucine could be underlined as the ‘first limiting amino acids’, but all other AASs were over ‘1’, in accordance with reference amino acid contents of FAO/WHO (>1.00), showing that farmed axillary seabream provides high nutritional quality and can be considered as a favourable protein source. The ratios of ∑EAA/∑AA (44%–46%) and ∑EAA/∑NEAA (79%–86%) exceeded the minimum recommendation of 40% and >60% by FAO/WHO for all three groups. It can be concluded that axillary seabream either farmed, farm‐aggregated or distant wild fish presented high‐quality protein generating a healthy source for human food.     

First detection of Edwardsiella ictaluri (Proteobacteria: Enterobacteriaceae) in wild Australian catfish

The bacterium Edwardsiella ictaluri is considered to be one of the most significant pathogens of farmed catfish in the United States of America and has also caused mortalities in farmed and wild fishes in many other parts of the world. E. ictaluri is not believed to be present in wild fish populations in Australia, although it has previously been detected in imported ornamental fishes held in quarantine facilities. In an attempt to confirm freedom from the bacterium in Australian native fishes, we undertook a risk‐based survey of wild catfishes from 15 sites across northern Australia. E. ictaluri was detected by selective culturing, followed by DNA testing, in Wet Tropics tandan (Tandanus tropicanus) from the Tully River, at a prevalence of 0.40 (95% CI 0.21–0.61). The bacterium was not found in fishes sampled from any of the other 14 sites. This is the first report of E. ictaluri in wild fishes in Australia.     

Freshwater nucleated pearl quality is influenced by host mussel growth traits in Hyriopsis cumingii

Hyriopsis cumingii is one of the most important freshwater pearl mussels in China. Recently, this species can produce freshwater nucleated pearls of high quality. Here, we investigated whether nucleated pearl quality is influenced by the growth traits of the host mussel or other factors like cultivation period. We implanted host mussels with a spherical nucleus consisting of a small piece of mantle tissue from donor mussels. After 24 and 36 months of culture, host mussel growth traits including body weight and shell length, height, width and weight were recorded. These factors were then correlated with the quality traits of the pearls they produced, such as nacre thickness, size, weight, lustre and colour. Results indicated pearls obtained at 36 months after seeding were significantly larger in terms of nacre thickness, size and weight compared to those harvested at 24 months. In particular, nacre thickness (r = 0.33–0.48, P = 0.00), pearl size (r = 0.39–0.43, P = 0.00) and pearl weight (r = 0.35–0.47, P = 0.00) were showed to be significantly correlated with host mussels shell length, body weight and shell weight at 24 or 36 months. Larger and heavier host mussels tended to produce bigger pearls. In contrast, host mussels did not affect pearl colour. Cumulatively, our results suggest that longer culturing times and a larger host mussel may help produce better quality nucleated pearl. This information can help guide selective breeding programs designed to improve pearl quality produced by H. cumingii.     

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.     

Experimental early pathogenesis of Streptococcus agalactiae infection in red tilapia Oreochromis spp.

Streptococcus agalactiae causes a severe systemic disease in fish, and the routes of entry are still ill‐defined. To address this issue, two groups of 33 red tilapia Oreochromis spp. each of 10 g were orally infected with S. agalactiae (n = 30), and by immersion (n = 30), six individuals were control‐uninfected fish. Three tilapias were killed at each time point from 30 min to 96 h post‐inoculation (pi); controls were killed at 96 h. Samples from most tissues were examined by haematoxylin–eosin (H&E), indirect immunoperoxidase (IPI) and periodic acid‐Schiff; only intestine from fish infected by gavage was evaluated by transmission electron microscopy. The results of both experiments suggest that the main entry site of S. agalactiae in tilapia is the gastrointestinal epithelium; mucus seems to play an important defensive role, and environmental conditions may be an important predisposing factor for the infection.     

Invertebrate uptake of lipids in the vicinity of Atlantic salmon (Salmo salar) aquaculture sites in British Columbia

Samples of invertebrates, primary producers, seawater and cores were taken surrounding several aquaculture sites in coastal British Columbia, Canada. Analysis of dissolved organic carbon (DOC) showed a significant decrease (P = 0.027) over 1 km and an overall average of 290 ± 40 μM at 1 m depth. At 10 m, the decrease was not significant and DOC averaged 81 ± 3 μM. The essential fatty acid 22:6ω3 (docosahexaenoic acid, DHA) was significantly lower nearer to the farm sites for all mollusc taxa as well as for mussels alone (P < 0.01). However, mussel wet weight as well as the zooplankton fatty acid marker (ZFA; Σ 20:1ω9, 22:1ω11, 22:1ω9) and 20:1ω9 individually, decreased significantly with distance from the farms (P < 0.03). Principal components analysis showed a similar trend with DHA being higher in molluscs further from the farm. Mussels had significantly higher amounts of DHA compared with other molluscs indicating their potential in multi‐trophic, co‐culturing systems. Bacterial fatty acid markers increased in molluscs with distance from the farms; however, 18:1ω7, which is representative of methane‐oxidizing bacteria, remained higher closer to the farm. In addition, DHA proportions in mussels showed a significant breakpoint at 339 m from the farm suggesting co‐cultured mussels would benefit from placement further than 339 m from farms where DHA would be optimized.     

Genetic assessment of milkfish (Chanos chanos Forsskal) stocks based on novel short tandem repeats for marker‐aided broodstock management

Milkfish hatchery broodstock are either from on‐grown wild‐caught or hatchery‐produced fry/juveniles. To determine if a marker‐assisted management scheme can be formulated for improved milkfish hatchery production, milkfish stocks were genetically characterized using nine novel short tandem repeats or microsatellites. Eight wild‐bred Philippine stocks (CLA, CUR, CAM, SIH, SBH‐I1, HH, PAL and ZH‐P₀), four hatchery‐bred stocks (SBH‐I2, SBH‐D, BoH and ZH‐F₁), two farm stocks of known mixed lineages (SPH and BDH) and one Indonesian hatchery‐bred stock (WJH) were assessed. WJH was included since milkfish fingerlings from Indonesia reared in Philippine farms could be developed into future broodstock. Mean allelic richness (Ar) was highest in wild‐bred stocks (9.5) and lowest in hatchery‐bred spawners (9.1). Mean expected heterozygosities (He) were relatively similar in all stocks with wild‐bred stocks slightly higher (0.67) than the others. An analysis of molecular variance indicated significant yet low genetic differentiation among stocks (F_ST = 0.013; p = .000) where variation (98.6%) was explained by intra‐stock differences. In some of the domesticated stocks, reductions in mean allelic richness were observed in first generation hatchery broodstock (e.g. ZH‐F₁; Ar = 8.3), compared with their founder stock (e.g. ZH‐P₀; Ar = 9.4). The Indonesian stock was similar to local wild‐bred stocks based on genetic variability indices; thus, it might be likely that the local stocks’ fitness traits could be comparable with the imported milkfish stock which has been perceived to be better. The quality of locally available farmed milkfish and prospects of formulating a broodstock management scheme for the production of good quality milkfish seedstock are herewith discussed.     

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.     

Estimation of gut contents of freshwater mussel, Lamellidens marginalis L.

This study examined the food organisms found in the gut of freshwater mussels, Lamellidens marginalis L. A total of 34 taxa of food organisms were recorded, out of which 30 taxa belonged to phytoplankton and four taxa to zooplankton. Both the groups comprised three families each: Cyanophyceae (blue‐green algae), Chlorophycea (green algae) and Bacillariophyceae (diatoms) constituted phytoplankton, whereas Euglenaceae, Chlamydodontidae and Brachionidae belonged to zooplankton. The total number of frequencies of phytoplankton (n=537) were almost nine times more than that of zooplankton (n=60). When blue‐green algae, green algae, diatoms and zooplankton (all the three families together) were tested for significant difference (P<0.05) following Duncan's multiple range test, the result showed only two groups. Blue‐green algae, green algae and zooplankton were not significantly different (P<0.05), forming group A, whereas diatoms were significantly different from others, forming group B. The present study showed that the maximum number of organisms that mussels feed upon belong to green algae, followed by diatoms, blue‐green algae and few taxa of zooplankton.     

Polymorphisms of the growth hormone gene in domesticated red sea bream populations (Pagrus major) based on minisatellite genotypes and nucleotide sequences

The genetic diversity of the growth hormone gene in domesticated red sea bream (pmaGH) was evaluated using a minisatellite DNA marker located in intron 3 (pmaGH22) and nucleotide sequences. The number of alleles of pmaGH22 was largely decreased in domesticated strains of red sea bream, and the possibility of selection pressures was also detected based on the analysis of the Hardy–Weinberg equilibrium in some strains. However, each strain inherited a small number of alleles of pmaGH22, and the entire domesticated population (combining all strains) showed a large number of alleles (n = 17), similar to the allelic richness of the wild population (n = 18.5). Based on nucleotide sequencing analysis, three synonym mutations were found in the coding regions, and also several SNPs and indels were found in the noncoding regions. In addition, four genealogies of growth hormone haplotypes were identified based on principal coordinate analysis, and these genealogies of pmaGH partly reflected allele size ranges of pmaGH22. Several haplotypes shared alleles of pmaGH22, and also fragment size homoplasy in pmaGH22 was suspected. These alleles of pmaGH22 and the haplotypes will be a useful indicator for divergence of pmaGH and for broodstock individual selection with minimum inbreeding effect.     

The Three‐spined Stickleback,Gasterosteus aculeatus Linnaeus 1758, plays a minor role as a host of Lepeophtheirus salmonis (Krøyer 1837) in the Gulf of Maine

The sea louse, Lepeophtheirus salmonis (Krøyer 1837), is a significant parasite of farmed salmon throughout the Northern Hemisphere. Management of on‐farm louse populations can be improved by understanding the role that wild fish play in sustaining and providing refuge for the local population of sea lice. In this study, 1,064 sticklebacks were captured. Of these animals, 176 individuals were carrying a total of 238 sea lice, yielding a prevalence and intensity of 16.5% and 1.4 lice per fish, respectively. Detailed examination of the sea lice on the three‐spined sticklebacks captured in Cobscook Bay found two L. salmonis individuals using three‐spined sticklebacks as hosts. A 2012 survey of wild fish in Cobscook Bay, Maine, found multiple wild hosts for Caligus elongatus (von Nordmann 1832), including three‐spined sticklebacks (Gasterosteus aculeatus L.), but no L. salmonis were found in this earlier study.     

A survey on Tetracapsuloides bryosalmonae infections in Slovene fresh waters

Slovenia has no history of health problems related to proliferative kidney disease (PKD) either in farmed or in wild fish. However, due to the past molecular evidence for the presence of Tetracapsuloides bryosalmonae DNA in tissues of some fish from open waters, a survey was conducted on wild salmonids that were primarily sampled for other purposes. In winter 2010–2011, specimens from a total of 244 rainbow trout, Oncorhynchus mykiss (Walbaum), and brown trout, Salmo trutta L., from 30 bodies of fresh water were examined for T. bryosalmonae using a PCR method. The sampled fish showed no clinical signs or gross pathological lesions characteristic of PKD. Nineteen (7.8%) fish from seven (23.3%) fresh waters were positive for T. bryosalmonae. The identity of PCR amplicons was confirmed by sequencing. With one exception, all the positive fish were found in waters from the regions where the average yearly temperatures and the environmental pollution are higher. This preliminary countrywide survey provided the first insight into the situation regarding T. bryosalmonae infection of wild salmonids in Slovenia.     

Substrate type and brightness preference of Schizothorax wangchiachii and Percocypris pingi juveniles

It is advantageous for fish to choose different substrate types and brightness based on their current life cycle stage, foraging requirements, predator avoidance and water conditions. We determined the substrate type and brightness preference of hatchery‐reared and wild juvenile Schizothorax wangchiachii and Percocypris pingi under laboratory conditions. Individuals and groups were exposed to four kinds of substrate types (sand: diameter < 1 mm, small gravel: diameter approximately 2–3 cm, large gravel: diameter approximately 10–15 cm, sand and medium gravel mix: diameter approximately 4–7 cm) and two kinds of substrate brightness (black and white). The results showed that hatchery‐reared and wild S. wangchiachii and P. pingi significantly preferred large gravel and black substrates regardless of the number of fish (p < .05). P. pingi had a significantly higher preference for black substrate than S. wangchiachii both in hatchery‐reared and wild individuals (p < .05). Hatchery‐reared and wild individuals of the two species shared a preference for large gravel and black substrates suggesting that substrate preferences might be genetically based. The findings in the present study could be used to improve the rearing conditions for the two species in hatchery and thereafter to enhance its adaptability after releasing to the wild.     

Dietary oils influence ovary and carcass composition and embryonic development of zebrafish

To evaluate the effect of different dietary lipids on zebrafish reproduction, we examined parameters including ovary and carcass composition, oestradiol (E2) levels, eclosion rate and embryonic development. In our study, zebrafish were subjected to a 5‐month feeding trial whereby olive (OLV), linseed (LIN), fish (FIS) or corn (CRN) oil were used to compose four isonitrogenous and isoenergetic diets. A positive correlation was found between ovary EPA and whole‐body E2 concentration (P = 0.005). The developmental dynamics was affected by dietary lipids and embryos from CRN treatment females; which had higher percentage of ARA in ovary (1.48 ± 0.44%) (P = 0.015), developed faster at 8–9 h postfertilization (hpf) than the ones originated from females receiving other diets (P = 0.0069). This effect was not sustained during later observation periods, suggesting ARA may act as a modulator of developmental dynamics only during initial phases. There was no clear effect of dietary lipid source on ovary protein content (P = 0.304) and eclosion rates at 72 hpf (P = 0.0623). Dietary fatty acids play an important role in reproductive outcome; however, additional studies are necessary to elucidate the mechanisms by which HUFA affect embryonic development.     

Effect of Arthrospira supplementation on Oreochromis niloticus gut microbiota and flesh quality

Supplementing fish feed with Arthrospira platensis has several beneficial effects, such as improved growth, survival and feed conversion, but little is known about how it affects gut microbiota or flesh quality. In this trial, feed with 3% A. platensis was given to Oreochromis niloticus (SPR, spirulina supplemented) over a period of 50 days and their gut microbiota and flesh quality were compared with controls (CTRs, no supplementation). Average daily growth was higher in SPR fish (p < 0.05). Regarding microbiota, a total of 5,470,272 readings (individuals) were obtained from 16 gut samples (n = 8 CTR, n = 8 SPR), with a gamma diversity of 1,563 species but no significant differences were found between treatments. Among the high inter‐individual variability found, some individual microbiota were dominated by Proteobacteria (36.6%) and Fusobacteria (32.9%) and others more by Actinobacteria (37%). Regarding flesh quality, significant differences were observed in flesh colour post‐mortem (a*, b* and chroma) (p < 0.05), with higher reflectance in the range of 470–500 and 620–670 nm in SPR tilapia, which corresponds with the reflectance range of β‐carotene and phycocyanin. However, rigor mortis was resolved earlier in SPR fish (p < 0.05), which suggests a shorter shelf life. Adding 3% of A. platensis in O. niloticus diet can improve growth and flesh colour and does not appear to modify intestinal microbiota.     

Cryopreservation of sperm in farmed blacklip abalone (Haliotis rubra Leach, 1814)

This study developed a technique of sperm cryopreservation using liquid nitrogen (LN) vapour in farmed blacklip abalone Haliotis rubra through evaluating the following five key factors: (1) cryoprotectant agent (CPA) toxicity; (2) cooling temperature; (3) thawing temperature; (4) sperm to egg ratio and (5) sugar addition, using sperm motility or fertilization rate as quality assessment indicators. The results demonstrated that 6% dimethyl sulfoxide (DMSO) was the best single CPA for sperm cryopreservation in this species. The highest post‐thaw sperm motility was achieved when sperm were exposed to LN vapour for 10 min at 5.2 cm above the LN surface and thawed at 60°C and recovered at 16°C in seawater baths. Post‐thaw sperm motility was found to be significantly higher when 6% DMSO was used in combination with 1% or 2% glucose than 6% DMSO alone. Further evaluation of fertilization rate between these CPAs showed that 6% DMSO+2% glucose achieved the highest fertilization rate of 70% at a sperm to egg ratio of 10 000:1.     

Incidence and timing of wild and escaped farmed Atlantic salmon (Salmo salar) in Norwegian rivers inferred from video surveillance monitoring

Run timing of escaped farmed Atlantic salmon Salmo salar vs. wild fish was compared by the use of video camera surveillance in 15 rivers over several years, covering 1600 km of the Norwegian coastline (from 58°N to 69°N). Annual runs of wild salmon varied among rivers from <200 fish to more than 10 000. During the surveillance period that for most rivers extended from late May to early October, larger‐sized salmon (fish ≥ 65 cm) generally entered the rivers earlier than small fish. The percentage of salmon identified as escaped farmed fish ranged from 0.1% to 17% across rivers with an average of 4.3%. Estimates of escapees are, however, assumed to represent minimum values because an unknown number of farmed fish passing the video cameras may have been misclassified as wild fish. By the use of a linear mixed model and generalised additive mixed models, it was found that the relationship between run timing and fish length differed significantly between farmed and wild salmon. While small‐sized farmed and wild fish (<65 cm) entered the river at about the same time, wild large salmon returned on average 1–2 weeks earlier than similarly sized escapees. The proportion of large‐sized farmed escapees also increased until late August and decreased thereafter. In contrast, there was a relatively constant and lower proportion of small‐sized escapees throughout the season. Within the surveillance period, there was no evidence of any exceptionally late runs of fish classified as escaped farmed salmon.     

Ultrastructural and biomolecular detection of Rickettsiales‐like organisms in tissues of rainbow trout with Red Mark Syndrome

Red mark syndrome (RMS) and US strawberry disease (US SD) are skin disorders affecting rainbow trout farmed in Europe and USA. The disease etiology has not yet been established. In spite of specific investigations, identifying Rickettsia‐like organism (RLO)‐ and Midichloria‐like organism (MLO)‐related DNA in affected individuals, these pathogens have never been observed. We performed histological, ultrastructural and biomolecular analysis on skin and spleen samples of trout with RMS. Examination by TEM revealed the presence of intracytoplasmic microorganisms resembling Rickettsiales within macrophages, fibroblasts and erythrocytes. The microorganisms were oval or short rod shaped (400–800 nm in length and 100–200 nm in width) and often showed a cell wall similar to Gram‐negative bacteria. PCR analysis for Rickettsiales supported these findings: 53% of affected trout were positive by both PCR and TEM The primers RiFCfw‐RiFCrev were used to anneal both the RLO 16S DNA sequence and the MLO 16S DNA sequence. For this reason, and in agreement with previous studies confirming the presence of Rickettsiales‐related DNA in trout with RMS, we assume that TEM detected microorganisms morphologically consistent with bacteria belonging to Rickettsiales order and could be considered as possible causative agents of RMS.     

Growth and survival of olive barb,Puntius sarana (Hamilton 1822) larvae produced with cryopreserved versus fresh sperm

Despite the success in fertilization and hatching of fish eggs with cryopreserved sperm, report on growth and survival of larvae produced from frozen‐thawed sperm is inadequate. The study evaluates the applicability of cryopreserved sperm for mass seed production by comparing the growth and survival of a popular food‐fish olive barb, Puntius sarana (Hamilton 1822) larvae produced from cryopreserved and fresh sperm. The eggs were artificially fertilized with cryopreserved and freshly collected sperm, and the growth and survival of produced larvae from both group recorded up to 12 weeks. The independent sample t‐test statistic showed the difference in lengths, t(718) = 0.241; P = 0.810 and weights, t(718) = 0.412; P = 0.680 were insignificant between two groups. There was also no significant difference, t(718) = ?0.758, P = 0.448 in survival of larvae produced from cryopreserved and freshly collected sperm. The study indicates that larvae of olive barb produced from cryopreserved sperm are equally compatible in growth and survival as the larvae produced from fresh sperm. Therefore, cryopreserved sperm can be applied for artificial fertilization of P. sarana to supply quality seed for aquaculture.