Dietary supplementation with the microalgae Parietochloris incisa increases survival and stress resistance in guppy (Poecilia reticulata) fry

Dietary enrichments with the arachidonic acid (ARA)‐rich microalga, Parietochloris incisa, on the survival of guppy (Poecilia reticulata) fry were examined. Diets were applied via Artemia enrichment to fish from two commercial farms for 34 and 36 days of experimental period (trials 1 and 2, respectively). In trial 1, Artemia nauplii were enriched with dry biomass of whole algal cells at 0 (control), 0.1, 0.2 and 0.4 mg mL^?1. Fry fed with Artemia enriched with 0.4 mg mL^?1 demonstrated the lowest mortality rates (24% and 1% in farms 1 and 2, respectively) compared with controls (36% and 13% in farms 1 and 2, respectively). In trial 2, fry were fed with Artemia, enriched with whole algal cells (0.4 mg ml^?1), algal hexane extract (HE; containing primarily ARA‐rich triacylglycerols and β‐carotene; 0.19 mg ml^?1) or the extraction residue (0.28 mg ml^?1). Acute stress (5 min air exposure) was applied after 18 days. The lowest mortality was recorded in the whole alga‐fed group (av. 26% and 2.6% in farms 1 and 2, respectively), with a slightly, but not significantly higher mortality in the HE‐fed group (av. 29% and 6.2% in farms 1 and 2, respectively). Elevated lysozyme was associated with the reduced mortality. Overall, the use of P. incisa as a dietary supplement for guppy fry during their first month of life enhanced their survival and stress resistance.     

Immunomodulation of rainbow trout (Oncorhynchus mykiss) fry by bath exposure to a β‐glucan from Euglena gracilis

Early developmental stages of fish mostly depend on innate immune factors for their protection. Augmenting these factors by application of different immunostimulatory substances may be beneficial for rearing and survival of the early life stages of fish. Bath administration of stimulants leads to a uniform exposure of fish independent of feed intake and reduces the individual handling. The present study demonstrates the immunostimulatory effect of β‐glucan (bath exposure) in rainbow trout fry at different dosages and exposure time. Rainbow trout fry (avg. wt. 770 mg; 87 days post hatch) were exposed to three different concentrations of β‐glucan (10, 100 and 1000 μg mL^?1) by bath exposure for 1 and 24 h. Expression of immune related genes from pooled internal organ samples of individual fish were analysed using a real time qPCR assay. Expression of complement factors (C3 and factor B) and acute phase proteins (hepcidin, precerebellin and transferrin) was significantly up‐regulated after 24 h bath stimulation with β‐glucan (100 μg mL^?1). These innate immune factors may play a vital role in clearance of pathogens. The expression of most of genes showed both a dose‐ and time‐dependent response. A medium dose (100 μg mL^?1) induced a significant increase in expression of complement factors and acute phase proteins mainly at 24 h exposure, whereas the highest dose of β‐glucan (1000 μg mL^?1) down‐regulated the expression of most of the studied genes. The result from the present study indicates that β‐glucan bath exposure could be applied for enhancing the innate immune factors even in fry.     

In vitro effect of levamisole on the cell viability,phagocytosis and respiratory burst of Barbel chub (Squaliobarbus curriculus) macrophages

The present study was to determine in vitro effect of levamisole on the immune functions of Barbel chub (Squaliobarbus curriculus), head–kidney‐derived (HKM), spleen‐derived (SM) and peripheral blood monocyte‐derived macrophage (PBM). Macrophages were incubated with levamisole 10³, 10¹, 10⁻¹ and 10⁻³ ng mL⁻¹ to assay the cell viability, respiratory burst and phagocytosis. The results showed that macrophages treated with levamisole 10⁻³ ng mL⁻¹ gave a maximum respiratory burst response, whereas levamisole 10³ ng mL⁻¹ had no effect. Phagocytosis activity of the macrophages treated with levamisole enhanced significantly when compared to control, maximum response being at 10⁻³ ng mL⁻¹. While using the methylthiazoletetrazolium method to measure the cell activity for 12, 24, 36, 48, 60 h, there was no significant role on proliferation and the cell viability began to decline after 24 h. In conclusion, levamisole is a potent enhancer of Barbel chub macrophage activity with low concentration.     

Effects of stocking density and algal concentration on the survival,growth and metamorphosis of Bobu Ivory shell,Babylonia formosae habei (Neogastropoda: Buccinidae) larvae

Independent and combined effects of stocking density and algal concentration on the survival, growth and metamorphosis of the Bobu Ivory shell Babylonia formosae habei larvae were assessed using a 5 × 5 factorial design with densities of 0.25, 0.5, 0.75, 1.00 and 1.50 larvae mL⁻¹ and algal concentrations of 5, 10, 15, 20 and 25 × 10⁴ cells mL⁻¹ in the laboratory. Larval growth, survival and metamorphosis were significantly affected by both the independent effects of stocking density and algal concentration and by their interaction. The highest per cent survival (72.5%) and metamorphosis (49.5%), fastest growth (41.57 μm day⁻¹) and shortest time to initial metamorphosis (10 days) all occurred at the lowest stocking density and the highest algal concentration. Both crowding and food limitation had independently negative impacts on the survival, growth and metamorphosis of larvae, and these negative impacts were further strengthened by the interaction of a higher stocking density and a lower algal concentration. Moreover, the results suggest that stocking density and algal concentration obviously played different roles in determining larval survival and growth. To maximize survival and growth, B. formosae habei larvae should be reared at a lower stoking density of 0.25 larvae mL⁻¹ and fed a higher algal concentration of 25 × 10⁴ cells mL⁻¹ in large-scale hatchery seed culture.     

The effect of β‐1,3‐glucan derived from Euglena gracilis (Algamune™) on the innate immunological responses of Nile tilapia (Oreochromis niloticus L.)

Algamune^? is a commercial additive produced from Euglena gracilis, providing a rich source of the β‐1,3‐glucan paramylon. Isolated kidney phagocytes of Nile tilapia were incubated with graded doses (0, 8, 16, 32, 64 and 128 μg/ml) of Algamune^? and purified paramylon to gauge their ability to elicit the production of reactive oxygen species. A linear response was observed for extracellular superoxide anion for both sources but only Algamune^? for intracellular superoxide anion. After corroborating the immunostimulant properties ex vivo, a feeding trial was conducted to evaluate the dietary supplementation of Algamune^? (0, 100, 200, 400 and 800 mg/kg of diet) for Nile tilapia. Fish were fed for 3 weeks, after which, fish were sampled for blood and head kidney phagocytes. The remaining fish were challenged with Streptococcus iniae. Macrophage extracellular superoxide anion production was significantly elevated in fish fed diets with 200 mg of Algamune^? kg^?1 when compared to fish fed the basal diet. Even though the disease challenge did not show statistical differences, it is worth mentioning that fish fed intermediate doses of Algamune^? had lowest numerical mortality values. Therefore, Algamune^? was demonstrated to enhance some immunological responses of tilapia both in ex vivo and in vivo.     

Mass culture of fairy shrimp Streptocephalus proboscideus (Crustacea – Anostraca) and its use in larviculture of the Persian sturgeon, Acipenser persicus

This study was conducted with cysts of Streptocephalus proboscideus obtained from the University of Gent‐Belgium. The cysts were hatched in Environmental Protection Agency (EPA) medium. The nauplii were reared at the Sturgeon Research Institute using a pure culture of Scenedesmus obliquus alga supplied at a density of 5 × 10³ cell mL⁻¹ that gradually increased to 1 × 10⁴, 5 × 10⁴ and 1 × 10⁵ cell mL⁻¹ with the growth of the nauplii. The nauplii attained sexual maturity and started producing cysts in 8 days and yielded a mean cyst number of 220±40 female⁻¹ brood⁻¹ cysts. These cysts were used in the larviculture of Persian sturgeon, Acipenser persicus (Borodin). Forty‐three larvae of Persian sturgeon (mean weight: 15.4±1.1 mg; mean length: 27.1±2.7 mm) with roughly absorbed yolk sacs were stocked in three aquaria and fed S. proboscideus nauplii at 8‐h intervals. By the end of the experiment (day 5), the mean weight and length of Persian sturgeon larvae were 51.4±13.3 mg and 20.7±1.4 mm respectively.     

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.     

Improved techniques for rearing mud crab Scylla paramamosain (Estampador 1949) larvae

A series of rearing trials in small 1 L cones and large tanks of 30–100 L were carried out to develop optimal rearing techniques for mud crab (Scylla paramamosain) larvae. Using water exchange (discontinuous partial water renewal or continuous treatment through biofiltration) and micro‐algae (Chlorella or Chaetoceros) supplementation (daily supplementation at 0.1–0.2 million cells mL⁻¹ or maintenance at 1–2 millions cells mL⁻¹), six different types of rearing systems were tried. The combination of a green‐water batch system for early stages and a recirculating system with micro‐algae supplementation for later stages resulted in the best overall performance of the crab larvae. No clear effects of crab stocking density (50–200 larvae L⁻¹) and rotifer (30–60 rotifers mL⁻¹) and Artemia density (10–20 L⁻¹) were observed. A stocking density of 100–150 zoea 1 (Z1) L⁻¹, combined with rotifer of 30–45 mL⁻¹ for early stages and Artemia feeding at 10–15 nauplii mL⁻¹ for Z3–Z5 seemed to produce the best performance of S. paramamosain larvae. Optimal rations for crab larvae should, however, be adjusted depending on the species, larval stage, larval status, prey size, rearing system and techniques. A practical feeding schedule could be to increase live food density from 30 to 45 rotifers mL⁻¹ from Z1 to Z2 and increase the number of Artemia nauplii mL⁻¹ from 10 to 15 from Z3 to Z5. Bacterial disease remains one of the key factors underlying the high mortality in the zoea stages. Further research to develop safe prophylactic treatments is therefore warranted. Combined with proper live food enrichment techniques, application of these findings has sustained a survival rate from Z1 to crab 1–2 stages in large rearing tanks of 10–15% (maximum 30%).     

The pigmenting effect of different carotenoids on fancy carp (Cyprinus carpio)

The optimal concentration of a panel of individual and combined carotenoid sources on skin pigmentation in fancy carp was investigated by nine experimental diets that were formulated and supplemented with astaxanthin at 25 mg kg^?1, lutein at 25 and 50 mg kg^?1, β‐carotene at 25, 50 and 75 mg kg^?1, and lutein combined with β‐carotene at 25 : 25 and 50 : 50 mg kg^?1, while a diet without supplemented carotenoid served as a control. The results showed that serum TC of fish fed diets containing supplemented with lutein plus β‐carotene at 25 : 25; 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet were higher than the other treatments (P ≤ 0.05). Serum TC of the respective treatments was 6.2 ± 2.0, 7.8 ± 3.3 and 7.3 ± 1.9 μg mL^?1 serum, respectively. Fish fed diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet had serum astaxanthin concentrations similar to fish fed the diet with astaxanthin alone at 25 mg kg^?1. Serum astaxanthin concentrations was 0.7 ± 0.01, 0.9 ± 0.01, 0.4 ± 0.02 and 1.7 ± 0.18 μg mL^?1 serum, respectively. The chromaticity of fish body skin of red and white position was assessed by colourimetry using the CIE L*a*b (CIELAB) system. Pigmentation response of skin redness of fancy carp fed with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were higher than other treatments (P ≤ 0.05) but they were similar to fish fed with 25 mg kg^?1 astaxanthin diet. The redness (a* values) of fish fed diets with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were 28.3 ± 0.53, 29.9 ± 1.38, 28.8 ± 3.95 and 28.5 ± 2.49, respectively. After 3 weeks of feeding the experimental diets, the fish fed on a diet without carotenoid supplement for one week demonstrated that the same three groups still retained their redness and had an overall tendency to improve skin colouring. Finally, concentrations 50 mg kg^?1 of lutein, or the combination of lutein and β‐carotene at 25 : 25 mg kg^?1 showed the highest efficiency for improving skin pigmentation and redness of skin.     

Effects of continuous and alternate administration of β‐glucan and mannan‐oligosaccharide on the growth,immunity and resistance against Vibrio splendidus of sea cucumber Apostichopus japonicus (Selenka)

A 4‐week growth trial was conducted to compare the effects of different feeding strategies of dietary immunostimulants on the growth, immunity and resistance against Vibrio splendidus of sea cucumbers Apostichopus japonicus (Selenka). Six feeding strategies were set, including feeding immunostimulants‐free diet continuously (control), feeding dietary β‐glucan (1.25 g kg^?1 diet) continuously, feeding dietary mannan‐oligosaccharides (MOS; 2.00 g kg^?1 diet) continuously, feeding β‐glucan 2 days followed by MOS 5 days alternately, feeding β‐glucan 5 days followed by MOS 2 days alternately and feeding β‐glucan 7 days followed by MOS 7 days alternately. The sea cucumbers fed immunostimulants showed higher specific growth rate (SGR) and lower cumulative mortality than control (P < 0.05). When sea cucumbers were fed with β‐glucan continuously, total coelomocytes counts and superoxide anion were significantly higher than control on the 4th day (P < 0.05). However, these two immune parameters were not significantly higher than those in control after the 18th day (P > 0.05). While sea cucumbers continuously fed MOS, these two immune parameters were not significantly higher than control until the 15th day. All immune parameters of the sea cucumbers fed with β‐glucan and MOS alternately were significantly higher than those in control during the experiment (P < 0.05). The sea cucumbers fed with β‐glucan 7 days followed MOS 7 days alternately showed the highest SGR and second lowest cumulative mortality. It was suggested that this feeding strategy is most suitable for sea cucumbers.     

Use of Diet Crossover to Determine the Effects of β‐glucan Supplementation on Immunity and Growth of Nile Tilapia,Oreochromis niloticus

Juvenile Nile tilapia were fed either a basal (control) diet (n = 6 aquaria) or a diet supplemented with 1 g/kg β‐glucan (n = 24 aquaria) for 4 wk. At the end of this period, fish receiving β‐glucan were continued on the same diet (n = 12 aquaria) or switched to the control diet (n = 12 aquaria) for 2 wk. After 6 wk, tilapia continuously fed the β‐glucan supplemented diets had improved weight gain and feed efficiency than those fed the control diet uninterrupted or switched from the β‐glucan diet to the control after 4 wk. Feeding tilapia β‐glucan for 4 wk and then switching to the basal diet for 2 wk caused a significant increase in the respiratory burst of polymorphonuclear lymphocytes (17.77 × 10³ units/1000 white blood cells [WBC]) compared to catfish fed the control diet (13.50 × 10³ units/1000 WBC) or the β‐glucan diet continuously (13.57 × 10³ units/1000 WBC), but other immune parameters were unaffected. Tilapia were then challenged with Streptococcus iniae. The two groups were divided again (n = 6 aquaria) postchallenge and continued on the same diet or switched to the other diet (β‐glucan or control) for another 3 wk. No differences in survival to S. iniae infection occurred between dietary groups.     

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.     

Fish antibiotherapy: bioencapsulation of flumequine using adult brine shrimp (Artemia salina)

Optimization of antibiotic delivery strategies to aquatic environment and to the specific characteristics of the target species is essential for the improvement of bacterial infection control measures. This work aimed at standardizing the use of Artemia salina to deliver flumequine to fish as antimicrobial treatment. Adult Artemia were used to bioencapsulate flumequine. A flumequine concentration of 358 μg mL⁻¹ was found adequate to perform bioencapsulation during 24 h without causing mortality. Antibiotic concentration in Artemia, quantified by means of a microbiological assay based on MIC determination, using Escherichia coli ATCC 25922 as control strain was 256.55 mg g⁻¹ (±71.22). The therapeutic doses of 10 mg kg⁻¹ BW, calculated on the basis of a consumption of about 4% BW/day, would then be delivered by the consumption of 7.8 Artemia g⁻¹ of fish.     

Bioassay‐guided isolation and identification of active compounds from Macleaya microcarpa (Maxim) Fedde against fish pathogenic bacteria

Four alkaloids (Sanguinarine, 6‐Methoxyl‐dihydro‐chelerythrine, Cryptopine and β‐Allocryptopine) were isolated from aerial parts of Macleaya microcarpa (Maxim) Fedde using bioassay‐guided isolation method, and the inhibitory activity of ethanolic extract, various fractions and these four alkaloids against four fish pathogenic bacteria (Aeromonas hydrophila, Aeromonas salmonicida, Vibrio anguillarum and Vibrio harveyi) was assessed in vitro using the agar dilution method and the microdilution assay method respectively. A. hydrophila was the most sensitive strain to all the tested compounds. Minimum inhibitory concentration (MIC) values were lower for sanguinarine against all tested Gram‐negative strains than other three alkaloids, with MIC values of 12.5 mg L^?1 for A. hydrophila and 50 mg L^?1 to other pathogenic bacteria. Followed by 6‐methoxyl‐dihydro‐chelerythrine, which showed considerable antibacterial activity with MIC values of 80 mg L^?1 for A. hydrophila, 100 mg L^?1 for V. harveyi, and 125 mg L^?1 for both V. anguillarum and A. salmonicida. Cryptopine and β‐allocryptopine revealed similar inhibitory activity with MIC values of 100 mg L^?1 for A. hydrophila and 200 mg L^?1 for other three bacterial species. These finding provided evidence that extract, as well as isolated compounds from M. microcarpa might be potential sources novel antibacterial agents for the treatment of fish infectious diseases.     

Effects of dietary β‐glucan and glycyrrhizin on non‐specific immunity and disease resistance of white shrimp,Litopenaeus vannamei (Boone) challenged with Vibrio alginolyticus

The white shrimp Litopenaeus vannamei, fed immunostimulant‐free, 0.2%β‐glucan and 0.06% glycyrrhizin diets for 18 days, respectively, were challenged with Vibrio alginolyticus at 6.4 × 10⁴ CFU shrimp^?1. The total haemocyte count (THC), phenoloxidase (PO) activity, respiratory burst (RB) and superoxide dismutase (SOD) activity changes for a 120‐h period were investigated, and shrimp mortality was also recorded. The results showed that PO activity, RB and SOD activity were significantly higher in shrimp fed the two immunostimulant diets after 18 days than those in shrimp fed immunostimulant‐free diets. The THC and SOD activity decreased significantly from 0 to 24 h post challenge, and then reverted to normal levels at 96 and 72 h respectively. The values for PO activity and RB increased from 0 to 48 h post challenge. Compared with those fed the control diets, shrimp fed immunostimulants had significantly higher PO activity and RB values at 120 h post challenge. Mortalities after challenge with V. alginolyticus were significantly lower in shrimp fed with β‐glucan or glycyrrhizin than in those fed with a diet without immunostimulants. It was concluded that dietary β‐glucan and glycyrrhizin increased the shrimp immunity. Furthermore, β‐glucan caused an increase in some immune parameters 12 h earlier than glycyrrhizin after V. alginolyticus challenge.     

Enhancement of the innate immune system and disease‐resistant activity in Cyprinus carpio by oral administration of β‐glucan and whole cell yeast

The effects of dietary β‐ (1,3) glucan and whole cell yeast (Sacharomyces uvarum) on the immune response and disease resistance to Aeromonas hydrophila were investigated in Cyprinus carpio. β‐(1,3) glucan was extracted from the yeast. Both β‐(1,3) glucan and whole yeast were incorporated into the diet at 1% level and fed to common carp C. carpio for a period of 60 days. Control and treated fish were exposed to A. hydrophila on the 30th and the 60th day of the experimental period. Dietary supplementation of glucan significantly increased the white blood cell count in fish on the 60th day (2.91±0.04 × 10⁴), and the highest nuetrophil nitro blue tetrazolium (NBT) activity was also observed in glucan‐fed fish (30th day). A consistent increase in neutrophil (NBT) activity was also observed in whole cell fed fish until the end of the experiment. Similarly, β‐(1,3) glucan and whole cell yeast enhanced the serum lysozyme activity from the 15th day onwards but higher activity was reported on the 30th day in glucan and the 60th day in whole cell yeast‐fed fish. Suplementation of β‐(1,3) glucan protected the fish from A. hydrophila infection. Nearly 75–80% of the fish survived pathogen exposure (relative percentage survival). However, only 54–60% survival was observed in the whole cell‐fed fish. β‐(1,3) glucan and whole cell yeast protect the fish from pathogens by enhancing the cellular and humoral immune response in C. carpio.     

Immunomodulatory effects of β‐glucans derived from Euglena gracilis or Saccharomyces cerevisiae for hybrid striped bass (Morone chrysops × M. saxatilis)

Beta‐glucans (BGs) can activate the animal's innate immune system, enhancing the primary defence lines against pathogenic insults. The objective of this study was to assess the immune responses of hybrid striped bass (HSB) when exposed to paramylon, derived from Euglena gracilis, or zymosan, derived from Saccharomyces cerevisiae. Two separate trials were conducted to evaluate the efficacy of these immunostimulants as a feed additive or injected intraperitoneally. Five diets were formulated to contain either paramylon or zymosan at two different levels (50 or 100 mg/kg), and no supplementation serving as the control. Blood and head kidney were sampled, and an elevated production of anion superoxide from isolated phagocytes was observed for all supplemented groups when compared to the control. Dietary paramylon at 50 mg/kg increased immunoglobulin levels in the plasma. The second trial was conducted by injecting a BG solution (10 mg of β‐glucan kg^‐1 of body weight) and phosphate buffer solution serving as the control. Seven days after injection, blood samples were collected and immunological profiles from whole blood and plasma were significantly (p < .05) affected by the treatments. The results from this study indicate that both dietary and injected paramylon and zymosan modulated the immunological responses of HSB.     

The effect of feeding the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on cell-mediated immunity and protection against Yersinia ruckeri in pikeperch (Sander lucioperca)

The present study examined the influence of leucine metabolite β‐hydroxy‐β‐methylbutyrate (HMB), a natural product HMB, on nonspecific cell‐mediated defence mechanisms and protection against enteric redmouth disease (ERM) in pikeperch (Sander lucioperca). β‐hydroxy‐β‐methylbutyrate was fed in a pelletted ration at 50 mg kg^?1 of the feed for 8 weeks. The phagocytic ability and potential killing activity of blood and pronephric phagocytes were examined in HMB‐ and control‐fed fish before and after 8 weeks of feeding HMB. Simultaneously, the proliferative response of blood and pronephric lymphocytes stimulated by mitogens concanavaline A and lipopolisaccharide were examined in experimental and control groups. Following 8 weeks of HMB feeding, a challenge test was performed by injecting the fish with live pathogenic bacteria Yersinia ruckeri. β‐hydroxy‐β‐methylbutyrate applied in the diet for 8 weeks prompted a statistically significant (P<0.05) increase in phagocytic ability and potential killing activity of the blood and pronephric phagocytes and the proliferative response of blood and pronephric lymphocytes. The changes in these mechanisms correlated with protection against Y. ruckeri, the ERM pathogen. The results showed that feeding HMB increased the nonspecific cell‐mediated defence mechanisms and protection against ERM by reducing cumulative mortality (30%) following the challenge with pathogenic bacteria. Future studies will include determination of optimal doses and protocols of oral application of HMB to maximize the immunomodulatory effects and protection against viral diseases in intensive pikeperch culture.     

Functional Properties of Ethyl Acetate-methanol Extract of Commonly Edible Molluscs

ABSTRACT

Marine molluscs are consumed as culinary delicacies and represent a relatively untapped source of natural functional food ingredients. The bioactive potential of ethyl acetate-methanol (C₂H₅OAc/CH₃OH) extract of molluscs Uroteuthis duvaucelii, Amphioctopus marginatus, Crassostrea madrasensis, and Sepiella inermis were demonstrated using different in vitro systems. The C₂H₅OAc/CH₃OH extract of A. marginatus displayed greater 2, 2-diphenyl-1-picryl-hydrazil (IC₉₀ 1.76 mg mL^?1) radical quenching capacity along with potential hydroxymethylglutaryl coenzyme-A reductase (HMGCR)-inhibiting activity (IC₉₀ 1.21 mg mL^?1) and angiotensin converting enzyme-I inhibitory property (ACE-I) (IC₉₀ 0.87 mg mL^?1) compared to those displayed by other molluscs. The organic extract of S. inermis set forth significantly higher pro-inflammatory 5-lipoxygenase inhibitory property (IC₉₀ 1.96 mg mL^?1) along with potential antidiabetic activity as determined by dipeptidyl peptidase-4 (DPP-4) antagonistic activities (IC₉₀ 2.62 mg mL^?1, P < .05). Significant correlations were observed between bioactive properties and electronegative groups of organic extracts of a mollusc. The free radical quenching property of the extracts exhibited a positive correlation towards other bioactivities, implicating the role of antioxidant property of the compounds in the extract to inhibit lifestyle ailments. The present study indicated that S. inermis and A. marginatus could be utilized as functional food ingredients to combat hypertension, hyperlipidemia, and inflammation-related risks.