Effect of <Emphasis Type="Italic">Lactococcus lactis</Emphasis> K-C2 on the growth performance,amino acid content and gut microflora of amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis>

This study aimed to evaluate the effect of Lactococcus lactis K-C2 on the growth performance, microbial diversity, and release of free amino acids in the intestinal tract and the edible parts of young amberjack, Seriola dumerili. Fish were fed a diet with or without strain K-C2 (2?×?10¹⁰ cfu/g feed) for 25 days. The results indicated that the growth performance of fish in the treated group was significantly higher than those in the control group (p?<?0.05). The amount of five amino acids (aspartate, sarcosine, taurine, alanine, and arginine) in the gut content and 13 of 21 amino acids in the edible parts of fish in the treated group were significantly higher (p?<?0.05) than those in the control group. Sphingomonas, Propionibacterium, and Mycobacterium were observed in gut microflora of fish in both the control and treated groups. Staphylococcus and Kocuria were detected in one sample from the control and treated groups; Acinetobacter and Acidobacteria were found in one sample from the control group. L. lactis was only found in one sample in the treated group. In conclusion, the dietary administration of probiotic L. lactis stimulated growth, reduced feed consumption, and improved the nutritional value of cultured amberjack.     

The immune modulatory effect of oregano (<Emphasis Type="Italic">Origanum vulgare</Emphasis> L.) essential oil on <Emphasis Type="Italic">Tilapia zillii</Emphasis> following intraperitoneal infection with <Emphasis Type="Italic">Vibrio anguillarum</Emphasis>

The current study aimed to evaluate the possible effect of Origanum essential oil on innate immune parameters as well as the hematological profiles of Tilapia zillii following challenge with Vibrio anguillarum. Fifty-four of Tilapia zillii weighing 180?±?10.2 g were randomly distributed into three identical closed recirculating seawater systems. The study included three groups (G1, G2, and G3) repeated in triplicates. Fish of the first two groups were fed on a basal diet without herbs, whereas fish of the last group were fed on a basal diet supplemented with Origanum essential oil at concentration 1 g kg^?1 for 15 days. Subsequently, fish of G2 and G3 subjected to a peritoneal inflammation by intraperitoneally injecting V. anguillarum (5.5?×?10⁵ CFU mL^?1), whereas fish of G1 injected with saline and served as control. Fish of all groups were then sampled at 4, 12, and 24 h post injection. No mortalities were observed in both basal and Origanum fed groups. However, some specimens of fish fed basal diet showed dorsal fin erosions, eroded mouth, and detached skin. Although the kinetics of RBCs, WBCs, Hb, and differential leukocyte values remained unchanged in fish fed different diets at the beginning of the trial, significant increases in those values were observed in fish fed Origanum essential oil at 12 and 24 h post injection. Similarly, an augmentation of plasma proteases, antiproteases, and lysozyme activities were recorded in fish fed Origanum essential oil at the same particular sampling points. A significant enhancement in plasma bactericidal capacity was only recorded in fish fed Origanum essential oil at 12 and 24 h post challenge compared to those fed basal diet. In conclusion, Origanum essential oil had a pronounced influence on the innate immunity and increased the fish resistance to V. anguillarum. These data gave insight into the potential use of Origanum in prophylactic strategies against threatening pathogens.     

Wheat grains fermented by fungal mycelia (<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> or <Emphasis Type="Italic">Lentinus edodes</Emphasis>) as alternative feed ingredients for juvenile rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

We investigated the effects of replacing non-fermented wheat grains with wheat grains fermented by fungal mycelia in the diet of juvenile rainbow trout (Oncorhynchus mykiss). We assessed growth performance, feeding parameters, and body composition in three experimental groups (0.33?±?0.01 g, in triplicates of 50 individuals each). The diets for all the groups contained ca. 43% protein and 19% lipids. Experimental diets were made by replacing the 100 g kg^?1 of wheat grains present in the basal diet (CTRL) with the same proportion of wheat grains fermented by Pleurotus ostreatus (PWD) or Lentinus edodes (LWD) mycelium. Fish were fed to apparent satiation twice a day for 56 days. Both, PWD and LWD, significantly increased fish body weight from day 28 onwards. Final body weight was 2.37?±?0.04 g (CTRL), 4.29?±?0.02 g (PWD), and 3.50?±?0.05 g (LWD), and feeding efficiency (%) was increased from 64.5?±?0.7 (CTRL) to 92.5?±?0.5 (PWD) and 84.8?±?1.5 (LWD). The experimental diets also improved nutrient retention efficiency (%): 30.0?±?0.5 (PWD), 27.7?±?1.1 (LWD), and 21.0?±?0.1 (CTRL), for crude protein; 40.3?±?0.6 (PWD), 31.0?±?1.8 (LWD), and 16.1?±?0.7 (CTRL), for ether extract; and 16.1?±?0.1 (PWD), 14.0?±?0.3 (LWD), and 11.6?±?0.6 (CTRL), for phosphorus. Body lipid content was highest for PWD followed by LWD and CTRL (81.4?±?1.4, 63.2?±?2.5, 42.3?±?2.6 g kg^?1, respectively), while viscerosomatic index was lowest for PWD (p?<?0.05). Liver glycogen in LWD and PWD fish (0.62?±?0.10 and 0.21?±?0.08% liver weight) was significantly higher than in CTRL fish (0.05?±?0.01% liver weight). Wheat-mycelium meals appear to be suitable dietary ingredients for improving juvenile rainbow trout growth and nutritional performance. These benefits vary according to the mushroom species used.     

Dietary supplementation of yeast (<Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>) improves growth,stress tolerance,and disease resistance in juvenile Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

The yeast Saccharomyces cerevisiae is one of the commonest probiotics incorporated in aquafeeds. An 84-day feeding trial was conducted to evaluate the effects of varying dietary inclusions of S. cerevisiae, 0% (control), 3% (YF3), 5% (YF5), and 7% (YF7), on growth, stress tolerance, and disease resistance in juvenile (body mass ~?21 g) Nile tilapia (Oreochromis niloticus). Fish were randomly distributed in groups of 20 into 12 1-m³ hapas and fed isoenergetic (~?17 kJ g^?1 gross energy) and isonitrogenous (~?300 g kg^?1 crude protein) diets at 3% of their bulk weight daily. Specific growth rates were significantly higher for the yeast-fed fish (0.77–0.78% day^?1) than for the control fish (0.60% day^?1) and resulted in significantly higher mean final weights for the yeast-treated groups. Protein and lipid retention efficiencies were also significantly higher in the yeast-fed fish than in the control group. In subsequent stress challenge trials, the yeast-fed fish had greater tolerance to acute heat as well as hypoxia exposure than the control fish. Survival rates of the yeast-treated groups following sudden exposure to elevated water temperature (40 °C) ranged from 82.5 to 100% compared to 15% for the control. Mean survival rates following a 24-h hypoxia exposure were also generally significantly higher for the yeast-fed fish. The probiotic groups recorded a relative percent survival (RPS) of 75% after a 14-day Aeromonas hydrophila infection challenge. The results of this study indicate that S. cerevisiae as an additive in Nile tilapia diets has beneficial impacts on growth, stress tolerance, and disease resistance.     

Demographic and competition studies on <Emphasis Type="Italic">Brachionus ibericus</Emphasis> and <Emphasis Type="Italic">Proales similis</Emphasis> in relation to salinity and algal (<Emphasis Type="Italic">Nannochloropsis oculata</Emphasis>) density

We isolated the rotifers Brachionus ibericus and Proales similis from the sediment of shrimp tanks and studied their individual demographic characters and competition between them at two food levels (0.25?×?10⁶, 1.00?×?10⁶ cells ml^?1 of Nannochloropsis oculata at 25 °C) and salinities ranging from 10 to 30‰. Our hypothesis was that growth rates would be higher with increasing food levels and salinities. Observations were taken twice a day for life table studies and daily once for population growth experiments. Using survivorship and fecundity data, we derived various life history variables. Although the average life span (7.6?±?0.4 days) and gross reproductive rate (33.8?±?2.9 neonate female^?1 day^?1) of B. ibericus were higher than those of P. similis (average life span 5.4?±?0.6 days and gross reproductive rate 13.0?±?0.6 neonate female^?1 day^?1), the population growth experiments showed that P. similis had higher r values (0.32?±?0.005 day^?1) than B. ibericus (0.23?±?0.002 day^?1) at 1.0?×?10⁶ cells ml^?1 of N. oculata. The rotifer P. similis was more adversely affected due to the presence of B. ibericus than vice versa. The data are important for developing techniques for a large-scale culture of these rotifers as food in aquaculture.     

Effects of exogenous non-starch polysaccharide-degrading enzymes in diets containing <Emphasis Type="Italic">Gracilaria lemaneiformis</Emphasis> on white-spotted snapper <Emphasis Type="Italic">Lutjanus stellatus</Emphasis> Akazaki

A feeding experiment was conducted to investigate the effects of exogenous non-starch polysaccharide (NSP)-degrading enzymes in diets containing Gracilaria lemaneiformis (GL) on growth performance and digestive enzyme activities of white-spotted snapper Lutjanus stellatus Akazaki (initial mass 8.0 ± 0.1 g). A basal diet (D0) containing a mixed protein source (fish meal, soybean meal and GL meal) was used as the control. Two diets supplemented with 0.5 g (D1) and 1 g (D2) exogenous NSP-degrading enzymes per kilogram of diet were formulated. Each diet was assigned to triplicate groups of fish in a total of nine floating sea cages (270 fish, 30 fish per cage). After a 60-day feeding trail, significantly higher weight gain, specific growth ratio and feed efficiency ratio were observed in fish fed D2 diet compared to those of control (P < 0.05). Body lipid, moisture and ash contents were not significantly affected by NSP-degrading enzyme supplementation, but significantly higher protein content was noticed in fish fed D2 diet compared to that of control (P < 0.05). The amylase activity in the stomach and intestine was significantly higher in fish fed D2 diet (P < 0.05), but no significant differences were observed in pepsin and lipase activities in the stomach or trypsin and lipase activities in the intestine between all treatments. The results suggested that addition of 1 g kg^?1 NSP-degrading enzymes in diet could efficiently improve seaweed feed utilization and growth performance of white-spotted snapper fish.     

In vitro and in vivo evaluation of probiotic properties of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> in Kenyi cichlid, <Emphasis Type="Italic">Maylandia lombardoi</Emphasis>

In this study, Enterobacter cloacae bacterium was isolated from curd and its antibacterial potential against the pathogen Plesiomonas shigelloides was evaluated using the freshwater ornamental fish Kenyi cichlid (Maylandia lombardoi). Among the bacterial isolates, E. cloacae exhibited tolerance to acidic pH 2 and demonstrated the highest antibacterial activity against P. shigelloides in various in vitro assays. Dietary supplementation of E. cloacae with prebiotic supplement 2% mannan oligosaccharide improved the growth performance and reduced the toxic metabolites such as nitrite in culture tank water of Kenyi cichlid. In vivo application of E. cloacae with mannan oligosaccharide significantly elevated (p?<?0.05) white blood cell counts (88.47?±?2.15 10³ mm^?3) and respiratory burst activity (0.243?±?0.007) of Kenyi cichlid when challenged with P. shigelloides. Intestinal morphology of Kenyi cichlid fish treated with probiotic and prebiotic combinations showed improved intestinal architectures. The present findings confirm that the isolated bacterium E. cloacae is a potential probiotic and can be used effectively to prevent infection of P. shigelloides in freshwater ornamental fish culture.     

<Emphasis Type="Italic">Curcuma longa</Emphasis> as additive in the diet for <Emphasis Type="Italic">Astyanax aff. bimaculatus</Emphasis>

The aim of the present study was to evaluate the effect of turmeric (Curcuma longa) as additive in the diet for Astyanax aff. bimaculatus. Fish (0.83 ± 0.04 g) were fed, for 60 days, with six diets containing 0.0, 20.0, 40.0, 60.0, 80.0, and 100.0 g turmeric kg^?1 feed. There was an increasing linear effect of turmeric on the thickness of the muscular layer, and height and width of the folds of the intestine. In the liver, a quadratic effect was observed of turmeric on the percentage of hepatocyte cytoplasm and a decreasing linear effect on the percentage of sinusoid capillaries. A quadratic effect was also observed of turmeric on the liver glycogen. There was no effect of turmeric on the antioxidant activity in the liver, carcass composition or productive performance of the fish. Thus, we concluded that Curcuma longa has trophic effects on the epithelium and the muscular layer of the intestine of A. aff. bimaculatus. Additionally, low levels of Curcuma longa cause increased deposition of liver glycogen and high levels cause reduction.     

Effects of potential probiotic <Emphasis Type="Italic">Lactococcus lactis</Emphasis> subsp. <Emphasis Type="Italic">lactis</Emphasis> on digestive enzymatic activities of live feed <Emphasis Type="Italic">Artemia franciscana</Emphasis>

In the present study, cellulase, protease, lipase and amylase activities were performed to investigate the effects of Lactococcus lactis subsp. lactis CF4MRS bioencapsulation of Artemia franciscana. Our results show that cellulase activities (total cellulase—FPase activity, exoglucanase and endoglucanase—CMCase activity) were significantly higher (P < 0.05) in A. franciscana tissue homogenates compared to those in the control group after 8 h of L. lactic bioencapsulation. Notably, an exception case was found in β-D-glucosidase activity, whereby the cellulase activity was not significantly different (P > 0.05) compared to the control group. Administrations of L. lactis at cell concentration of 10⁸ CFU mL^?1 showed considerable improvement of other important enzymatic activities such as amylase, protease and lipase in A. franciscana. The amylase/protease ratio in probiotic-treated A. franciscana was recorded at 0.343, approximately two times higher than those without probiotic administration (0.184). In contrary, amylase/lipase ratio showed half of a reduction (0.330) in L. lactis-administrated A. franciscana compared to the control (0.614). Our study suggests that important digestive enzymes, e.g., cellulase, amylase, protease and lipase, can be enhanced through bioencapsulation of A. franciscana with L. lactis subsp. lactis, which could in turn lead to further stimulation of endogenous enzymes in the fish and shrimp larvae.     

Fat deposition pattern and mechanism in response to dietary lipid levels in grass carp, <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>

This study aimed to evaluate the fat deposition pattern and lipid metabolic strategies of grass carp in response to dietary lipid levels. Five isonitrogenous diets (260 g kg^?1 crude protein) containing five dietary lipid levels (0, 20, 40, 60, 80 g kg^?1) were fed to quadruplicate groups of 15 fish with initial weight 200 g, for 8 weeks. The best growth performance and feed utilization was observed in fish fed with lipid level at 40 g kg^?1. MFI and adipose tissue lipid content increased with increasing dietary lipid level up to 40 g kg^?1, and higher lipid level in diet made no sense. Fish adapted to high lipid intake through integrated regulating mechanisms in several related tissues to maintain lipid homeostasis. In the present study, grass carp firstly increased PPARγ and CPT1 expressions in adipose tissue to elevate adipocyte differentiation and lipolysis to adapt to high lipid intake above 40 g kg^?1. In liver, fish elevated hepatic lipid uptake but depressed biosynthesis of hepatic FAs, resulted in no difference in HSI and liver lipid content among the groups. Only in muscle, fish showed a significant fat deposition when the lipid intake above 40 g kg^?1. The excess lipid, derived from enhanced serum TC and TG contents, was more likely to induce deposition in muscle rather than lipid uptake by adipose tissue in grass carp fed with high dietary lipid, indicating the muscle of grass carp might be the main responding organ to high lipid intake.     

<Emphasis Type="Italic">Virgibacillus proomii</Emphasis> and <Emphasis Type="Italic">Bacillus mojavensis</Emphasis> as probiotics in sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) larvae: effects on growth performance and digestive enzyme activities

This study examined the effects of two probiotics (Virgibacillus proomii and Bacillus mojavensis) on the digestive enzyme activity, survival and growth of Dicentrarchus labrax at various ontogenetic stages in three separate experiments. These probiotics were incorporated as single or mixed into fish feed for a period of 60 days. The growth parameters, proximate composition of whole body, digestive enzymes and gut microbiology were monitored at regular. The increments in length and weight and the survival were significantly higher (P < 0.05), and the values of food conversions were significantly lower (P < 0.05) in fishes fed the probiotic. The administration of V. proomii and B. mojavensis in diet resulted in an increase (P > 0.05) in body ash and protein content and in the specific activity of phosphatase alkaline and amylase in the digestive tract of all the fishes. V. proomii and B. mojavensis persisted in the fish intestine and in the feed in high numbers during the feeding period (group 1: 5.8 × 10⁴ CFU/ml, group 2: 9.6 × 10⁴ CFU/ml, and group 3: 9.8 × 10⁴ CFU/ml day 60). The two probiotics V. proomii and B. mojavensis were adequate for improved growth performance and survival and for healthy gut microenvironment of the host.     

Pre-sedation and transport of <Emphasis Type="Italic">Rhamdia quelen</Emphasis> in water containing essential oil of <Emphasis Type="Italic">Lippia alba</Emphasis>: metabolic and physiological responses

The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 µL/L of essential oil (EO) from Lippia alba leaves were investigated. Prior to transport, the fish in the two experimental groups were sedated with 200 µL/L of EO for 3 min. After transport, dissolved oxygen, carbon dioxide, alkalinity, water hardness, pH, temperature and un-ionized ammonia levels in the transport water did not differ significantly among the groups. However, total ammonia nitrogen levels and net Na⁺, Cl^? and K⁺ effluxes were significantly lower in the groups transported with EO of L. alba than those in the control group. PvO₂, PvCO₂ and HCO₃ ^? were higher after transporting fish in 40 µL/L of EO of L. alba, but there were no significant differences between groups regarding blood pH or hematocrit. Cortisol levels were significantly higher in fish transported in 30 µL/L of EO of L. alba compared to those of the control group. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) showed different responses after adding EO to the transport water. In conclusion, while the EO of L. alba is recommended for fish transport in the conditions tested in the present study because it was effective in reducing waterborne total ammonia levels and net ion loss, the higher hepatic oxidative stress in this species with the same EO concentrations reported by a previous study led us to conclude that the 10–20 µL/L concentration range of EO and lack of pre-sedation before transport are more effective.     

Growth and fatty acid composition of <Emphasis Type="Italic">Octopus vulgaris</Emphasis> paralarvae fed with enriched <Emphasis Type="Italic">Artemia</Emphasis> or co-fed with an inert diet

The rearing of Octopus vulgaris paralarvae during its planktonic life stage is a major challenge, as mortality is currently very high and unpredictable. In this study, we examined the survival and growth rates, as well as the fatty acid composition, of O. vulgaris paralarvae fed on three different dietary treatments: group ArDHA was offered juvenile Artemia enriched with a lipid emulsion (Easy DHA-Selco^®); group ArMA was fed with juvenile Artemia enriched with a mixture of microalgae (Rhodomonas lens and Isochrysis galbana); and group ArMA+ID received the same Artemia as group ArMA complemented with an inert diet. Dietary treatments were tested in triplicate with homogenous groups of paralarvae (25 individuals l^?1) established in 50-l tanks, and the experiment was conducted for 15 days. The survival rate of 15-day post-hatch (-dph) paralarvae from groups ArMA (20 ± 8%) and ArMA+ID (17 ± 4%) tended to be higher than in group ArDHA (13 ± 5%), though these differences were not statistically different. The dry weight (DW) of 15-dph paralarvae increased by almost 60% in groups ArMA and ArMA+ID, and nearly 40% in group ArDHA, with respect to hatchlings. The fatty acid (FA) composition of paralarvae revealed a remarkable drop of docosahexaenoic acid (22:6n-3, DHA) from hatchlings to 15-dph paralarvae of all groups (P < 0.05). However, paralarvae from group ArDHA contained higher levels of DHA than those from ArMA and ArMA+ID (P < 0.05). Despite Artemia enriched with DHA-Selco^® contained three-times more DHA than Artemia enriched with microalgae, no beneficial effects of this dietary treatment were observed on the performance of paralarvae.     

Dietary protein and carbohydrate requirement of juvenile Hawaiian limpet (<Emphasis Type="Italic">Cellana sandwicensis</Emphasis> Pease, 1861) fed practical diet

This study was conducted to evaluate dietary protein and carbohydrate requirement of juvenile Hawaiian limpets Cellana sandwincensis. A total of 64 juvenile limpets (3.12 ± 0.86 g) were fed five different dietary protein levels ranging from 270 to 470 g kg^?1 for 90 days. Carbohydrate and lipid levels were held constant at 180 and 49.7 g kg^?1, respectively. Weight gain and growth rates of the animals did not differ significantly (P > 0.05) among the protein levels ranging from 270 g kg^?1 (0.30 % day^?1) to 470 g kg^?1 (0.23 % day^?1). Next, opihi were fed four diets with protein levels from 210 to 500 g kg^?1 with a constant carbohydrate level at 120 g kg^?1. Weight gain and specific growth rates of opihi increased with increasing dietary protein from 210 to 350 g kg^?1, and significantly (P < 0.05) decreased at the 500 g kg^?1 diet. Highest weight gain, growth rates, and protein efficiency ratio were achieved at 350 g kg^?1. Elevated carbohydrate levels (180–370 g kg^?1) produced a significant difference (P < 0.05) in growth. The fastest growth rates of animals were obtained with 270 g kg^?1 (0.27 % day^?1) and 320 g kg^?1 (0.26 % day^?1). The weight gain of animals fed 180 and 370 g kg^?1 carbohydrate diets were significantly (P < 0.05) lower than those of animals fed 270 and 320 g kg^?1. We conclude that about 350 g kg^?1 protein and 320 g kg^?1 carbohydrate levels could be used for opihi.     

Modulation of appetite,lipid and glucose metabolism of juvenile grass carp (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>) by different dietary protein levels

This study was undertaken to explore the systemic metabolic strategies of juvenile grass carp (Ctenopharyngodon idellus) to maintain growth when fed with different dietary protein levels. The optimal growth group and two growing discomfort groups were selected through the basic data, to explain the growth difference from appetite regulation and lipid and glucose metabolism perspective. Three experimental diets were formulated with three dietary protein levels at 200.3, 296.1 and 442.9 g kg^?1, named P1, P2 and P3, respectively. Juvenile grass carp (initial body weight 12.28 ± 0.14 g) were fed with three diets with 3 replications per dietary treatment in an indoor recirculation system for an 8-week feeding trial. Fish fed with diet P2 dietary group showed significantly higher WG, SGR, FI and PER than other groups. Compared with other groups, mRNA expressions of NPY, Y8a and Y8b in fish fed with P2 significantly down-regulated, while the expressions of CCK and CART in fish fed with P3 significantly down-regulated (P < 0.05). With increasing dietary protein levels, G6Pase, GK, PK and PEPCK were all significantly inhibited (P < 0.05). For lipid metabolism, the mRNA expression of ACC in P1 dietary group was significantly higher than P3 dietary group; besides, LPL expression in P3 group was significantly higher than other two groups (P < 0.05). PPARα expression in P2 was significantly lower than other groups (P < 0.05). These results suggested that grass carp fed with P2 (296.1 g kg^?1 protein level) showed highest weight gain, contributed to more balanced nutrient metabolism and appetite regulation. Too high dietary protein (442.9 g kg^?1) should be avoided because it induced lowest PER, body lipid and liver lipid, and inhibited glucose and lipid metabolism in juvenile grass carp.     

Substitution of soybean meal with fermented <Emphasis Type="Italic">Jatropha</Emphasis> kernel meal: effect on growth performance,body composition,and metabolic enzyme activity of <Emphasis Type="Italic">Labeo rohita</Emphasis>

A 60-day feeding trial was conducted to evaluate the nutritional potential of fermented Jatropha kernel meal (FJKM) in the diets of rohu, Labeo rohita, fingerlings. Four iso-nitrogenous (336 g kg^?1) and iso-energetic (20 MJ kg^?1) diets containing 0, 100, 200, and 300 g kg^?1 FJKM in replacement of 0, 33.3, 66.7, and 100% soybean meal protein, respectively, were prepared and designated as control, T1, T2, and T3, respectively. A total of 180 acclimatized fish (average weight 6.05?±?0.04 g) were equally distributed into four experimental groups in triplicates, and were fed their respective diets to satiation twice daily at 10:00 h and 18:00 h. Higher percent weight gain (%WG); specific growth rate (SGR); protein efficiency ratio (PER); protease, aspartate amino transferase (AST), and alanine amino transferase (ALT) activities; and lower feed conversion ratio (FCR) were recorded in T2 and T3 groups compared to other groups. Whereas hepatosomatic index (HSI), intestinal somatic index (ISI), and amylase, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and catalase activities among the various dietary groups did not vary significantly (P?>?0.05). Whole body composition of fish analyzed at the end of the feeding trial exhibited significantly (P?<?0.05) higher ether extract and lower crude protein in the control group compared to the FJKM-fed groups. These results indicated that rohu fingerlings can efficiently utilize FJKM without any detrimental effect on growth performance, nutrient utilization, and metabolic response.     

Inclusion of <Emphasis Type="Italic">Pediococcus acidilactici</Emphasis> as probiotic candidate in diets for beluga (<Emphasis Type="Italic">Huso huso</Emphasis>) modifies biochemical parameters and improves immune functions

Administration of probiotic candidates in fish has generally been shown as a useful strategy to improve growth performance, survival, digestive enzyme activity, and gut microbiota. Unfortunately, the sero-immunological responses of different fish to different probiotic candidates are poorly understood. The present study assessed the effect of Pediococcus acidilactici as a probiotic on the biochemical and immunological parameters of beluga. Fish (248.32?±?10.21 g) were fed a control diet (without P. acidilactici( and three different doses of P. acidilactici-supplemented diets (10⁷, 10⁸, and 10⁹ CFUg^?1 diets) for 8 weeks. On week 8, blood and serum were sampled. Dose-dependent increase of immunological parameters (respiratory burst activity, lysozyme content, serum antibacterial activity, and total immunoglobulin) and biochemical parameters (total protein and albumin levels) was observed. However, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly increased in the juvenile beluga fed by 10⁹ CFUg^?1 P. acidilactici-supplemented diet compared to the other groups. Based on the results of this evaluation, it is reasonable to conclude that the inclusion of P. acidilactici as probiotic in diets for juvenile beluga improves the sero-immunological parameters of the fish and should be considered by farmers as a strategy to improve fish health.     

Influence of the dietary inclusion of <Emphasis Type="Italic">Gracilaria cornea</Emphasis> and <Emphasis Type="Italic">Ulva rigida</Emphasis> on the biodiversity of the intestinal microbiota of <Emphasis Type="Italic">Sparus aurata</Emphasis> juveniles

Partial substitutions of fish meal by 5, 15, or 25 % of Gracilaria cornea or Ulva rigida in experimental diets were evaluated to study their effects on biodiversity of intestinal microbiota composition in gilthead seabream (Sparus aurata) juveniles. The diets were offered to duplicate groups of 15 juvenile fish (14.0 ± 0.5 g) for 70 days, and at the end of the experiment the intestinal microbiota from four specimens of each treatment was analysed by denaturing gel gradient electrophoresis. Results showed that the substitution of fish meal by algae meal induced important modifications in the intestinal microbiota community, as a big reduction of the biodiversity when the highest percentage (25 %) of U. rigida was included. On the contrary, an increase on the number of species was detected when a 15 % of algae was included. Various Lactobacillus delbrueckii subspecies were selectively stimulated when G. cornea was included in the feed, and other bacterial species, such as those included in the Vibrio genus, were reduced.     

Cloning and characterization of <Emphasis Type="Italic">Bax1</Emphasis> and <Emphasis Type="Italic">Bax2</Emphasis> genes of <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> and evaluation of transcript expression in response to grass carp reovirus infection

Multidomain proapoptotic Bcl-2-associated X (Bax) protein is an essential effector responsible for mitochondrial outer membrane permeabilization, resulting in cell death via apoptosis. In this study, two Bax genes of grass carp (Ctenopharyngodon idellus), designated as CiBax1 and CiBax2, were isolated and analyzed. The obtained CiBax1 cDNA is 2058 bp long, with a 579 bp open reading frame (ORF) coding a protein of 192 amino acid residues. The full-length cDNA of CiBax2 is 1161 bp, with a 618 bp ORF coding 205 amino acids. Both CiBax1 and CiBax2 are typical members of Bcl-2 family containing conserved Bcl and C-terminal domains, and they share conserved synteny with zebrafish Bax genes despite the grass carp Bax mapping to different linkage groups. Phylogenetic analysis showed that CiBax1 was clustered with Bax from most teleost fish, and CiBax2 was close to Bax2 from teleost fish but far separated from that of Salmo salar. Quantitative real-time PCR analysis revealed broad expression of CiBax1 and CiBax2 in tissues from healthy grass carp, but the relative expression level differed. The mRNA expression of CiBax1 and CiBax2 was both upregulated significantly and peaked in all examined tissues at days 5 or 6 post-infection with grass carp reovirus. Subcellular localization indicated that CiBax1 protein was localized in both nucleus and cytosol, while CiBax2 protein only in cytosol. Moreover, CiBax2, but not CiBax1 was colocalized with mitochondrion under normal condition. Taken together, the findings would be helpful for further understanding of the function of Bax in teleost fish.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.