1,3‐1‐6 ß‐glucans enhance tissue regeneration in zebrafish (Danio rerio): Potential advantages for aquaculture applications

High aquacultural rearing density and handling of fish may frequently result in skin or gills wounds, thereby facilitating the onset of secondary infections. The capacity of the zebrafish to regenerate tissues, as well as fins and other organs, makes it an ideal animal model for studying the mechanisms of tissue regeneration. Since macrophages are involved in tissue regeneration, a diet including ß‐glucans might positively affect the process through activation of macrophages and other immune pathways. Consequently, the aim of this study was to investigate the effects of the oral administration of 1,3‐1,6 β‐glucans on the regeneration process of the caudal fin after its amputation in zebrafish. One hundred and twenty zebrafish were randomly distributed into four groups with three replicates each: an untreated non‐amputated group (CNA) and an untreated amputated group (CA) fed a control diet; two treated and amputated groups (MI and MII) fed for 14 days the same diet with the addition of two differently extracted 1,3‐1,6 ß‐glucans (MacroGard^® and Experimental MacroGard^®, Biorigin^©). ß‐glucans were added to allowed the administration of 12.5 mg/kg of fish body weight (0.35 g/kg of feed). Results showed that 1,3‐1,6 ß‐glucans decreased fish mortality rate and enhanced both daily and cumulative regenerated fin area, independent of the specific ß‐glucan extraction method used. Based on the mechanisms similarities of the innate immune system and tissue regeneration among different teleost species, these results may likely be extended to species of interest for the aquaculture sector.     

Inhibition of Vibrio spp. by 2‐Hydroxyethyl‐11‐hydroxyhexadec‐9‐enoate of Marine Cyanobacterium Leptolyngbya sp. LT19

Seven marine cyanobacteria were isolated from two regions of the Gulf of Thailand and evaluated by the agar diffusion method for antibacterial activity. Inhibitory compound was purified from the crude methanol extract and its structure was elucidated based on extensive spectroscopic analysis, including IR, 1D and 2D NMR spectra as well as mass spectrometry. A novel antimicrobial compound produced by the marine cyanobacterium Leptolyngbya sp. LT19 was identified to be a 2‐hydroxyethyl‐11‐hydroxyhexadec‐9‐enoate which has so far never been reported in microorganisms. Biological assays revealed that this novel compound exhibited antibacterial activities against the Gram‐negative, persistent shrimp pathogens, Vibrio harveyi and V. parahaemolyticus with minimal inhibitory concentration of 250–1000 and 350–1000 μg mL^?1 respectively.     

Development and validation of a real‐time PCR assay for the detection of Aeromonas salmonicida

A real‐time PCR assay using a molecular beacon was developed and validated to detect the vapA (surface array protein) gene in the fish pathogen, Aeromonas salmonicida. The assay had 100% analytical specificity and analytical sensitivities of 5 ± 0 fg (DNA), 2.2 × 10⁴ ± 1 × 10⁴ CFU g^?1 (without enrichment) and 40 ± 10 CFU g^?1 (with enrichment) in kidney tissue. The assay was highly repeatable and proved to be robust following equivalency testing using a different real‐time PCR platform. Following analytical validation, diagnostic specificity was determined using New Zealand farmed Chinook salmon, Oncorhynchus tshawytscha (Walbaum), (n = 750) and pink shubunkin, Carassius auratus (L.) (n = 157). The real‐time PCR was run in parallel with culture and all fish tested were found to be negative by both methods for A. salmonicida, resulting in 100% diagnostic specificity (95% confidence interval). The molecular beacon real‐time PCR system is specific, sensitive and a reproducible method for the detection of A. salmonicida. It can be used for diagnostic testing, health certification and active surveillance programmes.     

Response of the intestinal mucosal barrier of carp (Cyprinus carpio) to a bacterial challenge by Aeromonas hydrophila intubation after feeding with β‐1,3/1,6‐glucan

The effect of dietary β‐glucan on the bacterial community in the gut of common carp (Cyprinus carpio) was examined after oral application of Aeromonas hydrophila. Carp received either feed supplemented with 1% MacroGard^®, a β‐1,3/1,6‐glucan, or a β‐glucan‐free diet. Fourteen days after feeding, half of the carp from each group were intubated with 10⁹ colony‐forming units (CFU) of a pathogenic strain of A. hydrophila. Gut samples were taken 12 hr to 7 days after application and analysed using microbiological and molecular biological techniques (NGS, RT‐PCR‐DGGE). The reaction of the mucosa and the microbiota to an A. hydrophila intubation differed in carp fed with β‐glucan compared to carp from the control group. In β‐glucan fed carp, the total bacterial amount was lower but the number of bacterial species was higher. Bacterial composition was different for carp from both treatment groups. The number of mucin filled goblet cells was reduced in carp fed the β‐glucan diet. Mucus was obviously released from the goblet cells and was probably washed out of the gut together with high numbers of bacteria. This might be protective against pathogenic bacteria and, therefore, feeding with β‐glucan may provide protection against infections of the gut in carp.     

Experimental evidence on prevention of infection by the ectoparasitic protozoans Ichthyobodo salmonis and Trichodina truttae in juvenile chum salmon using ultraviolet disinfection of rearing water

In northern Japan, juvenile chum salmon Oncorhynchus keta (Walbaum) are released from hatcheries to enhance the fishery resource. Infections with ectoparasitic protozoans, particularly the flagellate Ichthyobodo salmonis and the ciliate Trichodina truttae, occasionally cause severe mortality among hatchery‐reared juveniles. This study examined the susceptibility of the two parasites to wide‐ranging UV irradiation (experiment 1) and then investigated whether UV disinfection of the rearing water using a commercial device was useful for preventing infections among juveniles in a small‐scale rearing system over a 28‐day period (experiment 2). In experiment 1, parasite mortality reached 100% with UV irradiation doses of ≥9.60 × 10⁵ μW s/cm² for I. salmonis and ≥8.40 × 10⁵ μW s/cm² for T. truttae. In experiment 2, disinfection of the rearing water at a UV irradiation dose of 2.2 × 10⁶ μW s/cm² succeeded in complete prevention of both parasites in the juvenile salmon. These results elucidate the minimum dose of UV irradiation for inactivation of I. salmonis and T. truttae, and demonstrate the usefulness of water disinfection using a commercial UV irradiation device to prevent infections by these parasites in hatchery‐reared juvenile chum salmon.     

Preliminary study on broodstock rearing,induced breeding and grow‐out culture of the sea cucumber Holothuria scabra in Sri Lanka

Experiments were conducted to identify suitable methods for broodstock rearing, induced breeding and grow‐out culture of Holothuria scabra in Sri Lanka. Two hundred and seventy‐two brooders (500–600 g) collected from off Mannar were individually packed in oxygen‐filled polythene bags with and without sea water and transported to a sea cucumber hatchery at Kalpitiya. Lagoon pens, sand‐filled fibreglass tanks and bare tanks were used in triplicates to maintain brooders. Spawning was initiated using air dry, water jet and thermal‐stimulation methods. Hatchery produced juveniles with an average weight of 11 ± 5 g were reared (2 individuals m^?2) in lagoon pens, mud ponds and fibreglass tanks in triplicates. The significantly high evisceration rate was observed when brooders were transported without sea water (t‐test, P < 0.05). Brooders maintained in bare tanks showed a significant weight reduction than the brooders in sand‐filled tanks and lagoon pens (anova , P < 0.05, d.f. = 2). Thermal stimulation (ambient temperature ± 3–5°C) was found to be the most successful method of spawning initiation of H. scabra. The mean (±SD) percentage males and females participated for spawning per trial was 9.2 (±10) and 4.6 (±5.6) respectively. On an average, 1.16 millions of eggs (±1.03 SD, n = 5) were obtained per spawning trial. H. scabra juveniles reared in tanks showed significantly lower growth rate than the juveniles in pens and ponds (anova , P < 0.05). Lagoon pens and sand‐filled tanks are suitable to maintain brooders and lagoon pens can be successfully used for mass rearing of juveniles.     

Synchronization of ovulation in cultured northern whitefish (Coregonus peled,Gmelin 1788) using [D‐Arg6Pro9Net]‐sGnRH analogue and its effect on egg quality

Female northern whitefish were treated with salmon [D‐Arg⁶Pro⁹Net]‐sGnRHa emulsified in Freund′s incomplete adjuvant as a sustained release treatment (sGnRHa‐FIA) or with [D‐Arg⁶Pro⁹Net]‐sGnRHa dissolved in physiological saline as acute release treatment (sGnRHa‐PS). Females in four experimental groups (A, B, C, D) and one control group (E) were injected intraperitoneally as follows: A and B – sGnRHa‐FIA at doses of 50 and 25 μg kg^?1, respectively; C – double injection (DI) of sGnRHa‐PS at 25 μg kg^?1 administered 3 days apart; D – single injection (SI) of sGnRHa‐PS at 25 μg kg^?1; and E – control group receiving a 1:1 mixture of FIA and physiological saline. All treatments induced and synchronized ovulation. Mean time to ovulation was significantly reduced (P < 0.05) in GnRHa‐treated groups compared with control. Hormone treatments did not affect the relative fecundity. Slight differences were found in ovarian fluid pH between group A and D (P < 0.05). Except for group C, egg size was significantly reduced (P < 0.01) in hormonally synchronized groups compared with the control. Survival to the eyed stage of eggs from females in groups A, B and C was significantly lower (P < 0.01) than in groups D and E. Per cent hatched alevins was lower (P < 0.01) in groups A, B and C than in groups D and E. We conclude that synchronization of ovulation in northern whitefish can be induced by a single acute injection of [D‐Arg⁶Pro⁹NEt]‐sGnRHa at 25 μg kg^?1 BW if given near the natural spawning time determined by water temperature below 2°C.     

Acute toxicity of nitrite on white shrimp Litopenaeus vannamei (Boone) juveniles in low‐salinity water

The nitrite toxicity was estimated in juveniles of L. vannamei. The 24, 48, 72 and 96 h LC₅₀ of nitrite‐N on juveniles were 8.1, 7.9, 6.8 and 5.7 mg L^?1 at 0.6 g L^?1; 14.4, 9.6 8.3 and 7.0 mg L^?1 at 1.0 g L^?1; 19.4, 15.4, 13.4 and 12.4 mg L^?1 at 2.0 g L^?1 of salinity respectively. The tolerance of juveniles to nitrite decreased at 96 h of exposure by 18.6% and 54.0%, when salinity declined from 1.0 to 0.6 g L^?1 and from 2.0 to 0.6 g L^?1 respectively. The safe concentrations at salinities of 0.6, 1.0 and 2.0 g L^?1 were 0.28, 0.35 and 0.62 mg L^?1 nitrite‐N respectively. The relationship between LC₅₀ (mg L^?1), salinity (S) (g L^?1) and exposure time (T) (h) was LC₅₀ = 8.4688 + 5.6764S – 0.0762T for salinities from 0.6 to 2.0 g L^?1 and for exposure times from 24 to 96 h; the relationship between survival (%) and nitrite‐N concentration (C) for salinity of 0.6–2.0 g L^?1, nitrite‐N concentrations of 0–40 mg L^?1 and exposure times from 0 to 96 h was as follows: survival (%) = 0.8442 + 0.1909S – 0.0038T – 0.0277C + 0.0008ST + 0.0001CT–0.0029SC, and the tentative equation for predicting the 96‐h LC₅₀ to salinities from 0.6 to 35 g L^?1 in L. vannamei juveniles (3.9–4.4 g) was 96‐h LC₅₀ = 0.2127 S² + 1.558S + 5.9868. For nitrite toxicity, it is shown that a small change in salinity of waters from 2.0 to 0.6 g L^?1 is more critical for L. vannamei than when wider differences in salinity occur in brackish and marine waters (15–35 g L^?1).     

Disruption of host‐seeking behaviour by the salmon louse,Lepeophtheirus salmonis,using botanically derived repellents

The potential for developing botanically derived natural products as novel feed‐through repellents for disrupting settlement of the salmon louse, Lepeophtheirus salmonis (Caligidae) upon farmed Atlantic salmon, Salmo salar, was investigated using an established laboratory vertical Y‐tube behavioural bioassay for assessing copepodid behaviour. Responses to artificial sea water conditioned with the odour of salmon, or to the known salmon‐derived kairomone component, α‐isophorone, in admixture with selected botanical materials previously known to interfere with invertebrate arthropod host location were recorded. Materials included oils extracted from garlic, Allium sativum (Amaryllidaceae), rosemary, Rosmarinus officinalis (Lamiaceae), lavender, Lavandula angustifolia (Lamiaceae), and bog myrtle, Myrica gale (Myricaceae), and individual components (diallyl sulphide and diallyl disulphide from garlic; allyl, propyl, butyl, 4‐pentenyl and 2‐phenylethyl isothiocyanate from plants in the Brassica genus). Removal of attraction to salmon‐conditioned water (SCW) or α‐isophorone was observed when listed materials were presented at extremely low parts per trillion (ppt), that is picograms per litre or 10^?12 level. Significant masking of attraction to SCW was observed at a level of 10 ppt for diallyl disulphide and diallyl sulphide, and allyl isothiocyanate and butyl isothiocyanate. The potential of very low concentrations of masking compounds to disrupt Le. salmonis copepodid settlement on a host fish has been demonstrated in vitro.     

The swimming capacity of juvenile Murray cod (Maccullochella peelii): an ambush predator endemic to the Murray‐Darling Basin,Australia

This study documented the swimming capacity of a large ambush predator, Murray cod Maccullochella peelii, endemic to the Murray‐Darling Basin, Australia. It was evident that the species is a swimming generalist, maintaining moderate ability across all aspects of the swimming capacity parameters that were investigated. For instance, the species was capable of prolonged swimming performance (critical swimming speed, U_crit: absolute, 0.26–0.60 m·s^?1, relative, 1.15–2.20 BL s^?1) that was inferior to active fish species, but comparable with other ambush predators. The species had low energetic demands, maintaining a low mass‐specific standard (21.3–140.3 mg·h^?1 kg^?1) and maximum active metabolic rate (75.5–563.8 mg·h^?1 kg^?1), which lead to a small scope for activity (maximum active metabolic rate–standard metabolic rate; 1.4–5.9). They were reasonably efficient swimmers (absolute and relative optimal swimming speed, 0.17–0.61 m·s^?1 and 0.77–1.93 BL·s^?1, respectively) and capable of repeat bouts of prolonged performance (recovery ratio = 0.99). Allometric changes in aspects of swimming capacity were realised with body mass, whereas broad swimming capacity was maintained across a wide range of temperatures. The swimming capacity demonstrated by M. peelii reflects a sit‐and‐wait foraging strategy that seeks to conserve energy characteristic of ambush predators, but with distinct features (e.g., lack of fast‐start ability) that may reflect their evolution in some of the world's most hydrologically and thermally variable rivers.     

Pre‐impoundment stock assessment of two Pimelodidae species caught by small‐scale fisheries in the Madeira River (Amazon Basin – Brazil)

The middle stretch of the Madeira River has supported an intensive commercial fishery for several decades. Developed in an area of rapids and waterfalls, this fishery focuses primarily on catfish stocks, mainly pimelodids. Data from fish landings collected in the years immediately prior to the start in operations of two large hydroelectric dams were used to estimate growth and mortality rates for Pirinampus pirinampu (Spix & Agassiz) and Brachyplatystoma platynemum Boulenger. In addition, stock assessment was carried out for both species, and the results used to evaluate the status of the fishery. Mean population parameters were L_∞ = 80.85 cm (total length), k = 0.52 year^?1, ? = 3.53, A_0.95 = 6 year, M = 0.87, F = 1.41–1.64 and Z = 2.28–2.51 for P. pirinampu; and L_∞ = 95.55 cm (total length), k = 0.28 year^?1, ? = 3.40, winter point = 0.41, A_0.95 = 11 year, M = 0.55, F = 0.16–0.35 and Z = 0.71–0.90 for B. platynemum. Yield‐per‐recruit analysis indicated P. pirinampu was overfished and that the stock of B. platynemum was under‐exploited. Moreover, overfishing of the P. pirinampu stock and the effects of fragmentation caused by the construction of the dam should be monitored.     

Density‐dependent effects of eastern Kamchatka pink salmon (Oncorhynchus gorbuscha) and Japanese chum salmon (O. keta) on age‐specific growth of western Alaska chum salmon

Hatcheries release >4.5 billion juvenile Pacific salmon (Oncorhynchus spp.) into the North Pacific Ocean annually, raising concerns about competition with wild salmon populations. We used retrospective scale analysis to investigate how the growth of chum salmon (O. keta) from western Alaska is affected by the abundance of chum salmon from Japanese hatcheries and wild pink salmon (O. gorbuscha) from the Russian Far East. Over nearly five decades, the growth of Kuskokwim River chum salmon was negatively correlated with the abundance of Japanese hatchery chum salmon after accounting for the effects of sex and spring/summer sea‐surface temperature in the Bering Sea. An effect of wild eastern Kamchatka pink salmon abundance on the growth of Kuskokwim River salmon was detectable but modest compared to the intraspecific competitive effect. A decrease in Japanese hatchery chum salmon releases in 2011–2013 was not associated with increased growth of Bering Sea chum salmon. However, the abundance of wild chum salmon from the Russian Far East increased during that time, possibly obscuring reduced competition with hatchery chum salmon. Our results support previous evidence that chum salmon are affected by intraspecific competition, and to a lesser extent interspecific competition, in the North Pacific, underscoring that the effects of salmon hatchery production transcend national boundaries.     

Purification and characterization of a β–glucan binding protein from the haemolymph of freshwater prawn Macrobrachium rosenbergii

β‐glucan binding protein (βGBP), a pattern recognition protein was purified from the haemolymph of freshwater prawn Macrobrachium rosenbergii by heparin affinity chromatography that showed a single band in native gradient PAGE. The β‐glucan binding property of the purified protein was confirmed in a phenoloxidase (PO) assay, where addition of βGBP along with β‐glucan increased the specific PO activity compared with that of β‐glucan alone. The molecular weight of the βGBP was found to be ~316 kDa on gel filtration chromatography. In SDS‐PAGE, βGBP molecule was reduced to one polypeptide chain of molecular weight ~113 kDa. Thus the βGBP in M. rosenbergii is possibly a homotrimeric molecule. The purified sample run on unreduced condition in SDS‐PAGE also revealed a similar size band (~113 kDa) and hence, the polypeptide chains of βGBP are held by non‐covalent interactions. The purified βGBP samples run in native PAGE was stained positively with alcian blue for carbohydrates and Sudan black for lipids indicating the βGBP to be a glycolipoprotein. With rabbit polyclonal anti‐βGBP serum developed, an indirect ELISA was standardized and the normal βGBP concentration in adult M. rosenbergii serum was quantified to be ~2 mg mL^?1. Furthermore, the applicability of the developed ELISA is discussed.     

The efficacy of MS‐222 as anaesthetic agent in four freshwater aquarium fish species

The efficacy of anaesthetic tricaine methanesulfonate (MS‐222) was evaluated in four freshwater aquarium fish species, Zebrafish (Danio rerio), Guppy (Poecilia reticulata), Discu (Symphysodon discus) and Green swordtail (Xiphophorus helleri). The correct dose of anaesthetic should induce the plane 4 of anaesthesia in less than 180 s, recovery in less than 300 s and must survive when exposed during 30 min to anaesthetic. Fishes were exposed to six concentrations of anaesthetic (75, 100, 125, 150, 200 and 250 mg L^?1) and the time of fish reaching plane 4 of anaesthesia, post exposure recovery, and the percentage of survival when fish were subject to 30 min in the anaesthetic were recorded. The optimal doses varied according to the species: D. rerio – 75, 100 and 125 mg L^?1, P. reticulata – 125, 150 and 200 mg L^?1, S. discus – 75 and 100 mg L^?1 and X. helleri – 125 and 150 mg L^?1. The induction time generally decreased significantly with increasing concentration of MS‐222 for all of the species evaluated. The recovery time had a tendency to increase with the increase of the MS‐222 concentration for D. rerio, P. reticulata and S. discus. On the other hand, X. helleri recovery time decreased with the increase of MS‐222 concentration. MS‐222 proved to be effective in anaesthesia for all the freshwater ornamental species studied. The main results clearly show that the optimal dose to anesthetize is fish species dependent and it is completely wrong to extrapolate optimal anaesthetic concentrations between different species.     

A3α‐peptidoglycan extracted from Bifidobacterium sp. could enhance immunological ability of Apostichopus japonicus

To assess the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the immune response and disease resistance of sea cucumber, different concentrations (0, 0.5, 5 and 50 mg mL^?1) of A3α‐PG suspensions were used to perform hypodermic injection on Apostichopus japonicus, followed by a Vibrio splendidus challenge. Total coelomocyte count (TCC), phagocytosis activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extra‐cellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), superoxide dismutase (SOD) and peroxidase (POD), were measured at 2, 6, 14 and 24 h post injection (hpi). The TCC was not significantly affected (P > 0.05) by A3α‐PG, ranging from 1.84 × 10⁶ to 3.53 × 10⁶ cells mL^?1. The coelomocyte phagocytosis activity was significantly activated (P < 0.05) in all the A3α‐PG treatments, whereas no significant difference was observed between them except 24 hpi (P > 0.05). The EC‐ACP activity in the 5.0 mg mL^?1 treatment increased significantly (P < 0.05) at all sampling times, while the IC‐ACP activity in the 50 mg mL^?1 treatment increased significantly (P < 0.05) at 2 hpi. Also, the 5.0 mg mL^?1 treatment had significant (P < 0.05) increase in the EC‐ALP activity within 14 hpi and the EC‐POD activity at 2 hpi, respectively, while significantly (P < 0.05) enhanced IC‐ALP and IC‐POD activities were observed in the 50 mg mL^?1 treatment within 6 hpi and at 2 hpi, respectively. Only the 5.0 mg mL^?1 treatment showed significant (P < 0.05) increase in the EC‐SOD activity at 2 hpi and IC‐SOD activity within 14 hpi, respectively. The challenge test showed that the animals treated with 50 mg mL^?1 of A3α‐PG had notably lower cumulative mortality after 14 days following V. splendidus exposure. All together, these results suggest that A3α‐PG could positively enhance immune response that effectively promotes the health status of A. japonicus against V. splendidus infection.     

Effects of stocking density on growth performance and welfare‐related physiological parameters of Atlantic salmon Salmo salar L. in recirculating aquaculture system

Stocking density, representing a potential source of long‐term stress, is widely recognized as a critical factor in aquaculture husbandry. Atlantic salmon were reared at low‐density (LSD, ~9.80–18.41 kg m^?3, initial to final density), medium‐density (MSD, ~19.62–36.96 kg m^?3) and high‐density (HSD, ~28.79–53.54 kg m^?3) stocking levels for 66 days to investigate the stress‐induced changes in fish growth and welfare. At the end of the trial, the salmon in HSD group showed significant lower final weight and higher ration level than those in MSD and LSD groups (P < 0.05). The salmon farmed in LSD group had higher special growth rate (SGR) and lower feed conversion ratio (FCR) (P < 0.05). However, stocking density could not affect the mortality, condition factor and coefficient of variation of salmon. The pectoral fin index of salmon in LSD group was significantly higher than that in HSD group (P < 0.05). Fifteen haematological and serum parameters were detected to assess the stress and welfare levels of salmon. The salmon farmed in HSD group had higher glucose (GLU) level and lower Cl^?, haemoglobin (HGB) and red blood cell (RBC) concentrations (P < 0.05). Contrast to the pattern of superoxide dismutase (SOD) activity, the cortisol, maleic dialdehyde (MDA) and alkaline phosphatase (ALP) levels of HSD group were higher than those in other two groups (P < 0.05). All these findings will provide a reference for selecting suitable stocking density in Atlantic salmon farming industry.     

Changing salmon: An analysis of body mass,abundance, survival,and productivity trends across 45 years in Puget Sound

Pacific salmon and trout (Oncorhynchus spp., Salmonidae) of the Puget Sound region of Washington State, USA, have experienced recent and longer‐term (multidecadal) variability in abundance while supporting robust fisheries. As part of the post‐season salmon management process, population‐specific estimates of total adult abundance to Puget Sound (Strait of Juan de Fuca) for pink (O. gorbuscha), chum (O. keta), coho (O. kisutch), sockeye (O. nerka), and Chinook (O. tshawytscha) salmon and steelhead trout (O. mykiss) are calculated annually. We compiled annual estimates of body mass, abundance and survival of hatchery‐ and naturally produced salmon from 1970 to 2015 to compare spatial and temporal patterns across species. Average weights of adult salmon and steelhead returning to Puget Sound, with the exception of coho salmon, have decreased since the 1970s. Temporal trends in abundance, survival and productivity varied by species and origin (hatchery vs. naturally produced). Generally, abundance and survival rates of natural‐origin species decreased whereas those of hatchery‐produced species did not, which is in contrast with other studies' general conclusions of decreasing survival among Puget Sound salmonids. Species diversity has decreased in recent years, with salmonids that rely on a short freshwater rearing phase in the natural environment (hatchery‐produced fish and naturally produced pink and chum) representing >90% of total returns in most years. This new information reveals patterns of body size, abundance, survival and productivity across species, life history and rearing type over the past 45 years and, in doing so, demonstrates the strength in multidecadal, multifactor time series to critically evaluate salmonid species.     

Effects of dietary live or heat‐inactivated autochthonous Bacillus pumilus SE5 on growth performance,immune responses and immune gene expression in grouper Epinephelus coioides

To evaluate the possible dietary application of live and heat‐inactivated probiotic Bacillus pumilus SE5 in grouper Epinephelus coioides, juveniles (14.6 ± 0.2 g) were fed either a basal control diet (without probiotic) or the basal diet supplemented with 1.0 × 10⁸ CFU g^?1 live (T1) and heat‐inactivated B. pumilus SE5 (T2). The heat‐inactivated probiotic significantly improved the final weight, weight gain (WG) and specific growth rate (SGR) at day 60 and significantly decreased the feed conversion ratio (FCR) at day 30 and 60, while the viable probiotic significantly decreased the FCR at day 60 (P < 0.05). Phagocytic activity, serum complement C3 and IgM levels as well as SOD activity elevated significantly in fish fed the heat‐inactivated probiotic for 60 days (P < 0.05). Furthermore, the heat‐inactivated probiotic remarkably up‐regulated expression of TLR2 and pro‐inflammatory cytokines (IL‐8 and IL‐1β) in head kidney (P < 0.05), but the viable probiotic failed to do so. These results indicated that heat‐inactivated B. pumilus SE5 can effectively improve the growth performance and immune responses of E. coioides.