Identification and florfenicol-treatment of pseudomonas putida infection in gilthead seabream (Sparus aurata) fed on tilapia-trash-feed.

The present study aimed to determine the major cause of the high mortality affecting farmed gilthead seabream (Sparus aurata) and controlling this disease condition. Fifteen diseased S. aurata were sampled from a private fish farm located at Eldeba Triangle, Damietta, fish showed external skin hemorrhages, and ulceration. Bacterial isolates retrieved from the diseased fish were identified biochemically as Pseudomonas putida and then confirmed by phylogenetic analysis of the 16 S rRNA gene sequence. P. putida was also isolated from three batches of tilapia-trash feed given to S. aurata. Biofilm and hemolytic assay indicated that all P. putida isolates produced biofilm, but 61.11% can haemolyse red blood cells. Based on the antibiotic susceptibility test results, P. putida was sensitive to florfenicol with minimum inhibitory concentrations ranging between 0.25 and 1.0 µg mL- 1, but all isolates were resistant to ampicillin and sulfamethoxazole-trimethoprim. Pathogenicity test revealed that P. putida isolate (recovered from the tilapia-trash feed) was virulent for S. aurata with LD50 equal to 4.67 × 107 colony forming unit (CFU) fish- 1. After intraperitoneal (IP) challenge, fish treated with 10 mg kg- 1 of florfenicol showed 16.7% mortality, while no mortality was recorded for the fish group that received 20 mg kg- 1. The non-treated fish group showed 46.7% mortality after bacterial challenge. HPLC analysis of serum florfenicol levels reached 1.07 and 2.52 µg mL- 1 at the 5th -day post-drug administration in the fish groups received 10 and 20 mg kg- 1, respectively. In conclusion, P. putida was responsible for the high mortality affecting cultured S. aurata, in-feed administration of florfenicol (20 mg kg- 1) effectively protected the challenged fish.

Dietary effects of microalga Tetraselmis suecica on growth, antioxidant-immune activity, inflammation cytokines, and resistance of Nile tilapia fingerlings to Aeromonas sobria infection.

The dietary effects of the green microalga Tetraselmis suecica (TS) on the growth, digestive enzymes, immune and antioxidant responses, genes expression, and disease resistance of Nile tilapia (Oreochromis niloticus) fingerlings were investigated. This microalga was mixed with the diet' ingredients at doses of 0.0 (the control), 5, 10, 15, and 20 g/kg diet and then fed to fish daily for 84 days. After the feeding trial, fish were experimentally challenged with Aeromonas sobria, infection and fish mortalities were recorded for another 10 days. Dietary TS significantly (p < 0.05) enhanced growth, digestive enzymes activities, and blood proteins, particularly at the level of 15 g/kg diet. Feeding the fish on 15 TS/kg feed exhibited highest mRNA expressions of GH and IGF-1 genes as well as SOD, CAT, and GPx genes compared to other TS groups. Moreover, highest levels of hepatic antioxidant and immune indices were found in the treatment of 15 g TS/kg feed. Significant downregulation of IL-1ß and IL-8 genes expression and significant upregulation of IL-10 gene expression were observed in TS-fed fish, principally in fish groups fed on 15-20 g TS/kg feed. Conversely, hepatic malondialdehyde levels, blood glucose, and the activities of transaminases (ALT and AST) were significantly (p < 0.05) decreased in fish fed with 15-20 g TS/kg diet. Serum bactericidal activity against A. sobria was significantly higher in TS-fed fish groups, and its highest levels were found in treatments of 15-20 g/kg diet. Of interest, the survival rates of fish groups fed diets with 10-20 g TS/kg feed were higher after the challenge with A. sobria infection than the control group. Accordingly, we can conclude that supplementing fish diets with a 15 g TS/kg diet enhanced the growth, antioxidant and immune activities, and resistance of Nile tilapia fingerlings to possible A. sobria infection.

Prophylactic effects of dietary caper (Capparis spinosa) extracts on the control of Streptococcus agalactiae infection, growth, immune-antioxidant, and inflammation cytokine responses of Nile tilapia fingerlings.

The antibacterial activity of aqueous (AE) or ethanolic extracts (EE) of caper (Capparis spinosa) against Streptococcus agalactiae was evaluated in vitro. Both caper extracts showed antagonistic activity against S. agalactiae and the inhibition zones in case of ethanolic extracts were larger than those of aqueous ones. Additionally, TEM investigations show that S. agalactiae cells treated with both C. spinosa extracts were damaged and degraded and this damage was greater in case of ethanolic extract. Another study was done to assess the promotion effects of dietary caper (C. spinosa) extracts on growth, antioxidant and immune activity, and inflammation cytokine responses of Nile tilapia (Oreochromis niloticus) and its resistance to S. agalactiae infection. However, fish (40 ± 2 g) were fed on diets containing 1.0 and 2.0 g/kg feed of each caper extract as well as the control group (free of caper) for 6 weeks. Fish were intraperitoneally injected (IP) with Streptococcus agalactiae at the end of the feeding trial, and fish mortality was tracked for additional ten days. Compared with other treatments, fish fed on 2.0 g EE/kg feed had higher counts of white and red blood cells as well as higher hemoglobin levels accompanied with lower AST and ALT activities. Antioxidant (superoxide dismutase and catalase activities) and immune, total protein, globulin, lysozyme, and immunoglobulin M) indices were increased along with significant decline in MDA levels in both caper extracts treated fish groups compared to the control group. Significant promotion in fish growth was affected positively with the increase in both caper extracts; particularly, the larger fish growth was observed in the treatment of 2.0 g EE/kg feed. Expressions of IL-1ß and IL-8 were declined; meanwhile levels of IL-10, SOD and CAT genes were upregulated in fish fed on 2.0 g EE/kg feed compared to other groups. After being challenged with S. agalactiae infection, fish survival was considerably (P < 0.05) greater in fish groups that fed on diets with caper extracts; particularly 2.0 g EE/kg feed (75%); while all fish fed on the control one were dead. According to these findings, the antioxidant and immune response of Nile tilapia fingerlings is stimulated by ethanolic extract of caper (2.0 g/kg feed), which also enhanced the growth performance and fish resistance to S. agalactiae infection.

Dietary Chlorella vulgaris effectively alleviates oxidative stress, immunosuppression, and enhances the resistance to Streptococcus agalactiae infection in cadmium-intoxicated Nile tilapia fingerlings.

Aquatic pollutants, including cadmium (Cd), cause oxidative stress on aquatic animals. The use of probiotics, including microalgae as a feed additive to alleviate the toxic impacts of heavy metals, is a much more interesting point. Hence, the current study investigated the oxidative stress and immunosuppression in Nile tilapia (Oreochromis niloticus) fingerlings caused by Cd toxicity as well as the preventive function of dietary Chlorella vulgaris against Cd toxicity. Accordingly, fish were fed on 0.0 (control), 5, and 15 g/kg diet of Chlorella up to satiation thrice a day, along with being exposed to 0.0 or 2.5 mg Cd/L for 60 days. Following the experimental procedure, fish from each group were intraperitoneally injected with Streptococcus agalactiae, and their survivability was observed for further ten days. Chlorella-supplemented diets meaningfully (P < 0.05) boosted the antioxidative capability of fish, which was evidenced by higher activities of hepatic superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) as well as higher levels of reduced glutathione (GSH) along with significant reductions in hepatic malondialdehyde levels. Moreover, the innate immunity indices [phagocytic activity (PA), respiratory burst activity (RBA), and alternative complement activity (ACH50)] were significantly higher in Chlorella-fed fish, particularly in the group of 15 g/kg diet. Additionally, serum of Chlorella-fed fish showed potent bactericidal activities against S. agalactiae, particularly at the treatment of a 15 g/kg diet. Feeding Chlorella diets to Nile tilapia fingerlings upregulated SOD, CAT, and GPx genes expression alongside the down-regulation of IL-1ß, IL-8, IL-10, TNF-α, and HSP70 genes expression. Conversely, Cd toxicity caused oxidative stress and suppressed the fish's innate immunity with upregulation of the expression of IL-1ß, IL-8, IL-10, TNF-α, and HSP70 genes. Feeding Cd-exposed fish on Chlorella-containing diets attenuated these adverse effects. The current research revealed that supplementing feeds with the treatment of 15 g/kg diet of C. vulgaris supports the antioxidant-immune responses and alleviates the Cd toxicity effects on Nile tilapia fingerlings.

Effects of dietary supplementation with microbial phytase on the growth, bone minerals, antioxidant status, innate immunity and disease resistance of African catfish fed on high soybean meal-based diets.

The microbial phytase, derived from Buttiauxella gaviniae, Yersinia mollarettiv and Hafnia spp., is proven to be safe for avian and porcine feeding and promotes their overall growth performance. Here, we have evaluated microbial phytase's effects on the growth, bone mineral content, antioxidant status, immune responses and the resistance of African catfish (Clarias gariepinus) fed with high soybean meal-based diets against Aeromonas hydrophila infection. Five isonitrogenous diets (40% protein) were supplemented with different levels of microbial phytase ranging from 0 as a control to 250, 500, 750 and 1000 FTU/kg diet. African catfish (n = 300; 8.5 ± 0.3 g) were allocated in 15 50-L tanks (in triplicates) and were fed on the prepared tested diets for 12 weeks. After the end of the feeding period of 12 weeks, 10 fish from each replicate was intraperitoneally infected with A. hydrophila (0.5 × 105 CFU/ml) and monitored for 14 days. Dietary phytase levels linearly and quadratically improved the growth performance of African catfish and stimulated feed intake. Bone levels of calcium, phosphorus, magnesium and zinc were also positively modulated in phytase-fed fish, especially at 750-1000 FTU/kg diet. Similarly, counts of red and white blood cells as well as haemoglobin, packed cells volume, platelets, lymphocytes and heterocytes were significantly modulated in all fish fed with phytase-supplemented diets. Higher levels of serum total protein, albumin and globulin were also observed in fish fed with a 750-1000 FTU/kg diet of phytase. Conversely, aspartate and alanine aminotransferase activities were lower in fish fed with a 750-1000 FTU/kg diet of phytase, compared to those fed the control diet. Moreover, antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase), reduced glutathione levels and immune responses (lysozyme, respiratory burst, protease and alkaline phosphatase activities) biomarkers were linearly and quadratically elevated, while malondialdehyde values were linearly and quadratically decreased in fish groups fed with phytase-based diets. After administering A. hydrophila, 60.0% of the fish fed the control diet perished, while no mortalities were observed in fish fed with 750-1000 FTU/kg diets. Taken together, the current study reveals that dietary phytase could improve the growth performance, blood profile, bone mineralization, antioxidant activities, immunity and overall protection of African catfish against A. hydrophila infection. Dietary phytase may be efficiently used in the feeding of African catfish to enhance their overall performance and mitigate health conditions with optimum level of 900 FTU/kg diet.

The growth performance, antioxidant and immune responses, and disease resistance of Litopenaeus vannamei fed on diets supplemented with Indian ginseng (Withania somnifera).

In the current study, white-leg shrimp (Litopenaeus vannamei) were fed on diets containing varying doses of Withania somnifera aqueous extract (WSAE) at a rate of 0 (control), 0.5, 1.0, and 2.0 g/kg feed for 56 days. After the feeding trial, shrimps in all groups were challenged with the exposure to Vibrio harveyi for ten days during which animals' mortality was observed. It is noted that the dietary WSAE linearly and quadratically stimulated shrimp's growth indices particularly at the treatment of 2.0 g/kg feed. Compared to the control group, the WSAE-fed L. vannamei had significantly higher villi length, villi width, and absorption area particularly in the treatment of 2.0 g/kg feed. Furthermore, L. vannamei fed on WSAE-enriched diets consumed more feed and exhibited higher total proteolytic activity, lipase, and α-amylase activities as compared with the control group. The dietary WSAE at escalating levels linearly and quadratically enhanced the antioxidant activity (serum superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), total antioxidant capacity, and reduced glutathione) and the immune response (total hemocyte counts, total protein, lysozyme, and phagocytic activity). Similarly, the mRNA expression levels of cMn-SOD, CAT, and GPx genes were linearly and quadratically upregulated in the hepatopancreas of L. vannamei fed on WSAE-enriched diets (especially in the 2.0 g/kg feed treatment), while their lowest levels were significantly observed in the control group. On the other hand, malondialdehyde levels were significantly decreased in WSAE-supplemented shrimp groups, and its highest levels were observed in animals fed on the control diet. After the bacterial exposure, the survival rates of L. vannamei fed on 1.0 and 2.0 g WSAE/kg feed (61.3% and 66.7%, respectively) were higher than those in the control animals. Taken together, the results obtained herein indicate that inclusion of WSAE in diets of L. vannamei effectively enhanced the growth, antioxidant biomarkers, immune response, and resistance to the V. harveyi infection, particularly at the treatment of 2.0 g/kg feed.

Growth performance, antioxidant activities, and immunological responses of hapa-reared thinlip mullet (Liza ramada) juveniles fed on diets supplemented with spirulina (Arthrospiraplatensis).

Spirulina (Arthrospira platensis) (SP) has been utilized for a long time as a valued feed supplement because of its proteinous content and other beneficial phytochemical compounds. Herein, we investigated the influences of SP-supplemented diets on growth, body somatic indices, digestive enzymes, hepatic antioxidant activities, and immunological responses of hapa-reared thinlip mullet (Liza ramada) juveniles. Fish were assigned in six triplicate groups and were fed for consecutive 60 days on the prepared experimental diets containing varying SP levels as 0.0, 2.0, 4.0, 6.0, 8.0, and 10.0 g/kg diet and defined as control (CNT or SP0), SP2, SP4, SP6, SP8, and SP10 groups, respectively. The results indicated that dietary SP supplementation linearly and quadratically improved the fish growth performance, and the highest growth indices were found in the SP8 group. However, dietary SP supplementation did not significantly alter feed conversion ratio (FCR), survival rate (%), hepato-somatic index, and viscera-somatic index among all experimental groups. Meanwhile, digestive enzymes (lipase, α-amylase, and proteases) in the mid-intestine were also linearly and quadratically increased in all SP-fed groups, and their uppermost values were noted in the SP8 group. Hepatic antioxidants such as superoxide dismutase, catalase, and total antioxidant capacity in SP-supplemented groups were significantly elevated than the CNT group. Conversely, hepatic malondialdehyde contents were decreased significantly along with increasing dietary SP-supplementation levels. The immunological parameters such as lysozyme, respiratory burst, and alternative complement activities were significantly elevated in SP-fed groups than in the CNT group. These findings evoked that feeding SP-supplemented diets (especially at 8.0 g/kg diet) significantly promoted the growth, digestive enzymes, hepatic antioxidant status, and immunity of L. ramada juveniles.

Dietary curcumin nanoparticles promoted the performance, antioxidant activity, and humoral immunity, and modulated the hepatic and intestinal histology of Nile tilapia fingerlings.

The current study investigated the effects of dietary curcumin nanoparticles (C-NPs) on the performance, hemato-biochemical profile, digestive enzymes activities, antioxidant status, humoral immunity, and liver and intestinal histology of Nile tilapia (Oreochromis niloticus). Fish (4.3 ± 0.5 g) were fed with diets enriched with 0.0 (control), 15, 30, 45, and 60 mg C-NPs/kg diet up to apparent satiety thrice a day for 60 days. The growth-stimulating effects of dietary C-NPs were significantly observed in terms of final weight, weight gain %, specific growth rate, and feed intake. Compared with the control group, serum amylase, lipase, and proteases activities of Nile tilapia significantly (P < 0.05) increased alongside the increase in dietary levels of C-NPs in a dose-dependent manner. The counts of red blood cells and white blood cells as well as hemoglobin and hematocrit levels of Nile tilapia fed with 30-60 mg C-NPs/kg diet were statistically (P < 0.05) higher than fish in the control group with no significant differences among them (P > 0.05). Moreover, lymphocytes and monocytes significantly (P > 0.05) increased; meanwhile neutrophils significantly (P > 0.05) decreased as C-NPs levels in diets increased. In a similar trend, antioxidant (malondialdehyde, superoxide dismutase, catalase, and glutathione peroxidase) and humoral immunity (lysozyme and total immunoglobulin) biomarkers were significantly higher in C-NPs-fed fish. Liver histology showed improvements in the cell architecture of fish fed with C-NPs containing diets up to 45 mg/kg diet. Compared with the control diet, feeding Nile tilapia with C-NPs diets resulted in a higher villi length/width and absorption area. According to the regression curves, the current study recommends using the dietary C-NP with optimum values of 45-55 mg/kg diet to improve the performance, digestive enzymes, antioxidant activities, and immunity response of Nile tilapia.

Impact of grape pomace flour (GPF) on immunity and immune-antioxidant-anti-inflammatory genes expression in Labeo rohita against Flavobacterium columnaris.

This study evaluates the effects of dietary inclusion of grape pomace flour (GPF) on growth, antioxidant, anti-inflammatory, innate-adaptive immunity, and immune genes expression in Labeo rohita against Flavobacterium columnaris. In both normal and challenged fish the growth rate, hematology and biochemical parameters significantly increased when fed with 200 and 300 mg GPF enriched diets; similarly the activities of antioxidants and innate-adaptive immune parameters, such as malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (GSH), phagocytic (PC), respiratory burst (RB), alternative pathway complement (ACP), lysozyme (Lyz), and total immunoglobulin M (IgM) significantly increased in both groups. Similarly, the immune, antioxidant, and anti-inflammatory-related gene mRNA expression was significantly up-regulated in head kidney (HK) tissues. The challenged fish fed without GPF always exhibited lower values of all the studied parameters. The results indicate that both normal and challenged fish treated with 200 mg GPF inclusion diet had significantly enhanced growth rate, antioxidant status, and immune defense mechanisms than with 300 mg GPF diet in L. rohita against F. columnaris.

Effect of diet enriched with Agaricus bisporus polysaccharides (ABPs) on antioxidant property, innate-adaptive immune response and pro-anti inflammatory genes expression in Ctenopharyngodon idella against Aeromonas hydrophila.

The effect of Agaricus bisporus polysaccharides (ABPs) supplemented diet on growth rate, antioxidant capacity, innate-adaptive immune response, proinflammatory and antiinflammatory genes expression in Ctenopharyngodon idella against Aeromonas hydrophila is reported. In both normal and challenged groups fed with 1.0 and 1.5 mg kg-1 ABPs diets resulted in a significant weight gain and feed intake. The survival was 100% in normal fish fed without or with any ABPs diet; the challenged fish fed with 1.0 mg kg-1 ABPs diet had 98.6% survival. The RBC and WBC counts, Hb, and Hct levels were significant in both normal and challenged groups fed with 1.0 and 1.5 mg kg-1 ABPs diets. A significant increase in total protein and albumin level was observed in both groups fed with 1.0 and 1.5 mg kg-1 ABPs diets. Significant increase in GPx, ROS, GR, GSH, PC, and MnSOD activity was observed in HK of both groups fed with 1.0 and 1.5 mg kg-1 ABPs diets; similarly both groups when fed with the same ABPs diets showed significant Lz, C3, and C4 activity. However, both groups fed with 1.0 mg kg-1 ABPs diet showed significant ß-defensin, LEAP-2A, IL-6, and NF-κB P65 mRNA expression. Similarly, IFN-γ2, IL-10, and TNFα mRNA expressions were significant in both groups fed with 1.0 mg kg-1 ABPs diet. The results indicate that both normal and challenged C. idella fed with a 1.0 mg kg-1 ABPs diet had better growth, antioxidant status, immune response, and pro-anti-inflammatory gene modulation against A. hydrophila.

Potential effects of dietary seaweeds mixture on the growth performance, antioxidant status, immunity response, and resistance of striped catfish (Pangasianodon hypophthalmus) against Aeromonas hydrophila infection.

Striped catfish (Pangasianodon hypophthalmus) is one of the important fish species in the world, which is widely cultured in several countries worldwide including Egypt. Aeromonas hydrophila infection showed a high prevalence in the farmed striped catfish negatively affecting its production causing economic loss. Therefore, the finding of functional feed supplements with antimicrobial activity is urgently needed. The use of marine seaweeds as feed supplements has produced satisfactory results as antibacterial agents. Accordingly, the present study was designed to evaluate the dietary effects of seaweeds mixture (Ulva lactuca, Jania rubens, and Pterocladia capillacea) extract (SME) on the growth performance, haemato-chemical parameters, antioxidant capacity, immune indices, and resistance of striped catfish fry against A. hydrophila infection. Four isonitrogenous (30% crude protein) diets were prepared to contain 0.0% (control), 1%, 2%, and 3% SME and were fed to striped catfish fry (1.7 ± 0.1 g) up to apparent satiety three times a day for 60 days. After the feeding trial, fish of each treatment were intraperitoneally injected with a virulent A. hydrophila strain and the relative percentage of survival (RPS) was recorded. The growth-stimulating activity of dietary SME was observed especially at 2% and 3% SME treatments. Similarly, haematological parameters (red blood cells, haemoglobin, haematocrit, and white blood cells) and serum protein profile (total protein, albumin, and globulin) were significantly (P < 0.05) improved, meanwhile serum alanine and aspartate aminotransferases were significantly (P < 0.05) decreased in fish fed with 2 and 3% SME diets. Serum creatinine and uric acid levels revealed no significant (P > 0.05) differences among the experimental groups. Hepatic malondialdehyde and serum nitrous acid levels were significantly decreased; meanwhile hepatic superoxide dismutase, catalase, and glutathione peroxidase levels were significantly increased in all SME-treated groups than the control one. Also, SME supplementations significantly increased the serum lysozyme, total immunoglobulin, and complement C3 activities with highest values at 2% and 3% SME treatments. Interestingly, after bacterial challenge, fish fed SME-supplemented diet were more resistant with the lowest fish mortality (15%) at the 3% SME diet, which exhibit the highest RPS (81.9%). Meanwhile the control group showed highest mortality (85%). According to the regression fitting curve, the current study recommends using the dietary SME with optimum levels of 2.25-2.5% to improve the growth performance, welfare status, and resistance of striped catfish fry against A. hydrophila infection.

Dietary inclusion of watermelon rind powder and Lactobacillus plantarum: Effects on Nile tilapia's growth, skin mucus and serum immunities, and disease resistance.

An eight-week investigation was conducted to access the potential impact of dietary watermelon rind powder (WMRP) and L. plantarum CR1T5 (LP) administered individually or in combination on immunity, disease resistance, and growth rate of Nile tilapia fingerlings cultured in a biofloc system. Three hundred twenty fish (average weight 16.57 ± 0.14 g) were distributed into 16 tanks at a rate of 20 fish per tank. The fish were fed different diets: Diet 1 (0 g kg-1 WMRP and 0 CFU g-1 L. plantarum) (control), Diet 2 (40 g kg-1 WMRP), Diet 3 (108 CFU g-1 LP), and Diet 4 (40 g kg-1 WMRP + 108 CFU g-1 LP) for eight weeks. A completely randomized design (CRD) with four replications was applied. Skin mucus, serum immunity, and growth parameters were analyzed every 4 weeks, and a challenge study against S. agalactiae was conducted at the end of the experiment. The findings showed that the inclusion of WMRP + LP, administrated individually or in a mixture, significantly (P<0.05) stimulated growth, skin mucus, and serum immune parameters of Nile tilapia fingerlings compared with the control. The highest values were detected in fish fed the combination of WMRP and LP, as opposed to individual administration of either WMRP or LP, in which no significant differences were detected. Within the challenge study, the relative percent survival (RPS) in Diet 2, Diet 3, and Diet 4 was 48.0%, 52.0%, and 68.0%, respectively. Fish fed 40 g kg-1 WMRP + LP produced significantly higher RPS and protection against S. agalactiae than the other treated groups. Current results suggest that the dual administration of WMRP and LP maybe an effective feed additive for Nile tilapia grown in an indoor biofloc system, capable of improving growth parameters and increasing resistance to S. agalactiae infection.

Dietary garlic and chitosan enhanced the antioxidant capacity, immunity, and modulated the transcription of HSP70 and Cytokine genes in Zearalenone-intoxicated European seabass.

The present study was performed to evaluate the toxic effects of feed-born zearalenone (ZEN) on antioxidative status, immunity, transcriptomic responses of European seabass, and the modulating roles of dietary garlic and/or chitosan powders. Fish (30.7 ± 0.6 g) were randomly arranged in five experimental groups (in triplicates), whereas the first group was fed on the control diet only without any supplements (control), and the second group was fed on the basal diet contaminated with ZEN (0.725 mg/kg diet). Three other groups were fed on ZEN-contaminated diets and simultaneously supplemented with garlic powder (GP) (30 g/kg diet) (ZEN + GP), chitosan powder (CH) (10 g/kg diet) (ZEN + CH), and a mixture of GP and CH (ZEN + GP + CH). Fish were fed on the experimental diets thrice a day for 4 weeks. Two-way ANOVA revealed a gradual decline in serum superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities in the ZEN group reaching their lowest levels at the 4th week. Contrariwise, malondialdehyde levels were markedly higher in the ZEN group reaching their highest values at the end of the experiment. A significant decline of total immunoglobulins (P < 0.05) was observed in the serum of the ZEN group, especially after the 4th week. Moreover, significant down-regulation of interleukin-4 (IL-4) and interleukin 1 beta (IL-1ß) genes (P < 0.05) alongside significant up-regulation of tumor necrosis factor-alpha (TNF-α) and heat shock protein 70 (HSP70) genes (P < 0.05) in the liver and anterior kidney of ZEN-intoxicated group. Interestingly, dietary supplementation with GP and CH significantly attenuated ZEN-induced oxidative stress, immunosuppression, and modulated transcriptomic responses of ZEN-exposed fish. Moreover, combined dietary supplementation of both feed additives resulted in better effects than each one alone.

Effects of sodium butyrate nanoparticles on the hemato-immunological indices, hepatic antioxidant capacity, and gene expression responses in Oreochromisniloticus.

Recently, nanotechnology has been greatly developed to provide the aquaculture industry with new beneficial nanomaterials to improve the health and welfare of aquatic animals. Herein, an eight-week experiment was designed to examine the dietary impacts of sodium butyrate nanoparticles (SB-NPs) on the hematological profile, blood proteins, immunological indices, antioxidant capacity, and expression analysis of cytokines and antioxidant-related genes in Oreochromis niloticus. Fish were randomly assigned into 5 experimental groups (3 replicates per group) and were fed diets supplemented with 5 levels of SB-NPs as 0.0 (control), 0.5, 1.0, 1.5, and 2.0 mg kg-1. The results revealed that supplementing diets with SB-NPs (1.0-2.0 mg kg-1) significantly elevated erythrocyte and leukocyte counts, hemoglobin concentrations, hematocrit values, total albumin, globulin, serum lysozyme activities, and total immunoglobulin M values compared with the control group. Notably, the highest levels of the parameters mentioned above were noticed in the group fed diet supplemented with 1.5 mg kg-1 SB-NPs. Moreover, dietary SB-NPs modulated the fish's antioxidant defense mechanisms, whereas there was a significant increase in hepatic superoxide dismutase, catalase, and glutathione peroxidase enzyme activities along with a significant decline in hepatic malondialdehyde concentrations in fish groups fed diets supplemented with SB-NPs (1.0-2.0 mg kg-1). A significant upregulation of antioxidant enzyme genes (gpx and sod), anti-inflammatory cytokine (il-10), and pro-inflammatory cytokines (il-1ß and il-8) were noticed in liver tissues of SB-NPs groups (0.5-1.5 mg kg-1). The highest mRNA expression folds of the above genes were recorded in the fish group fed diet supplemented with 1.5 mg kg-1 SB-NPs. In this context, we hypothesized that dietary supplementation with SB-NPs can boost the antioxidant status and immunity of O. niloticus. However, further research studies are still recommended.

Black soldier fly (Hermetia illucens) larvae meal in diets of European seabass: Effects on antioxidative capacity, non-specific immunity, transcriptomic responses, and resistance to the challenge with Vibrio alginolyticus.

Black soldier fly larvae meal (BSFLM) has been successfully demonstrated as a promising fish meal (FM) replacer in diets of several fish species including European seabass (Dicentrarchus labrax). However, its impacts on antioxidant capacity, and immune responses of treated fish are still poorly understood. A 60-day feeding trial was carried out to evaluate the effects of partial substitution of FM with different levels of dry BSFLM on the antioxidative status, non-specific immunity, transcriptomic responses, and resistance of European seabass to the challenge with Vibrio alginolyticus. Four isoproteic (45%) and isolipidic diets were formulated by replacing 0.0%, 25%, 35%, and 50% of the dietary FM. Each diet was randomly assigned to four fish groups (in triplicates) (initial mean body weight, 12.1 ± 0.21 g) (20 fish per aquarium) (n = 240). Fish were fed three times daily to the apparent satiation. At the end of the feeding trial, serum antioxidant biomarkers such as malondialdehyde levels, and catalase, superoxide dismutase, and glutathione peroxidase enzyme activities were significantly increased in all BSFLM groups in a dose-dependent manner compared to the control group (P < 0.05). The non-specific immune indices, including phagocytic activity, phagocytic index, serum lysozyme and respiratory burst activities were significantly elevated in BSFLM groups compared to those in the control group (P < 0.05). Significant upregulation of the mRNA expression levels of hepatic heat shock protein 70, interleukin-1beta and interleukin-10 genes were observed in all BSFLM groups compared to the control group (P < 0.05). Additionally, after the challenge with V. alginolyticus, the relative percent of survival was significantly elevated in fish groups fed on diets containing graded levels of BSFLM over the control group (P < 0.05). Conclusively, the present study suggests the potential efficacy of partial replacement of dietary FM protein for up to 50% by BSFLM without negative effects on fish health with possible potentiation of the antioxidative status, and the immune responses of the European seabass.

Dietary origanum essential oil improved antioxidative status, immune-related genes, and resistance of common carp (Cyprinus carpio L.) to Aeromonas hydrophila infection.

The use of herbal essential oils in aquafeeds is an important approach to maintain the fish health status. The origanum essential oil (OEO), extracted from Origanum vulgare, was included at levels of 0, 5, 10, 15, and 20 g per kg diet to evaluate its beneficial effects on antioxidant, immunity status, and expression of immune-related genes of common carp, Cyprinus carpio L. After 8 weeks, dietary inclusion of OEO significantly increased the activities of hepatic superoxide dismutase and catalase (P < 0.05); meanwhile, levels of hepatic malonaldehyde significantly decreased (P < 0.05). The levels of serum lysozyme activity, phagocytic activity, and phagocytic index have been linearly and quadratically increased by dietary OEO in a dose-dependent regime showing their highest levels in 15-20 g OEO/kg diet. Moreover, dietary OEO significantly upregulated the relative expression of hepatic interleukin-1beta (IL-1ß) and interleukin-10 (IL-10) genes (P < 0.05). Dietary OEO maintained the normal histomorphological criteria and density of melanomacrophage centers of spleens, and hematopoietic tissues of the anterior kidneys in all fish groups with respect to the control. After challenge with pathogenic bacterium Aeromonas hydrophila, the 10-day cumulative mortality significantly decreased (P < 0.05); meanwhile, the relative percent of survival of common carp significantly increased by OEO administration (P < 0.05) in a dose-dependent manner. The obtained results reflect the beneficial roles of dietary OEO to boost the antioxidative status and the immune responses in common carp with an optimum level of 15 g/kg diet.

Dietary taurine incorporation to high plant protein-based diets improved growth, biochemical, immunity, and antioxidants biomarkers of African catfish, Clarias gariepinus (B.).

Plant protein (PP) sources are generally used in high levels in fish diets. Mostly, PP sources are deficient in taurine; hence, there is a need for its supplementation to fish fed high PP diets. Therefore, effects of dietary taurine were examined on growth performance, feed utilization, immunity, and antioxidant parameters of African catfish, Clarias gariepinus (B.). Fish (10.3 ± 0.4 g) were fed on diets (40% crude protein) containing different taurine levels of 0 (control), 10, 20, 30, or 40 g/kg diet for 12 weeks. Fish fed a taurine-free diet (the control) with high PP sources showed poor growth as compared with these fed taurine-enriched diets where taurine stimulatory effects were observed on fish growth and feed intake. Feed conversion ratio and fish survival rate were not significantly differed among different treatments. Fish fed taurine-enriched diets showed also higher levels of serum glucose, cholesterol, total protein, albumin, globulin, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, urea, and creatinine over that fed the control diet. Furthermore, lysozyme and respiratory burst activities as well as superoxide dismutase and catalase activities were significantly elevated in fish fed taurine-enriched diets (P < 0.05) and their highest levels were observed in fish fed 30 g/kg diet. Additionally, taurine deposition in fish muscles was positively correlated with dietary taurine levels (P < 0.05). The present study concludes that taurine is a limiting factor for growth, immunity, and antioxidants responses of African catfish fed high PP-based diets and it should be incorporated in its diets with an optimum level of 20 g/kg diet.

Immunostimulatory effect of dietary chitosan nanoparticles on the performance of Nile tilapia, Oreochromis niloticus (L.).

Feed supplements to fish are generally used to overcome any expected diseases and stressors and to sustain eco-friendly fish farming. One of these feed supplements is chitosan, which stimulated growth and immune properties for many aquatic organisms. It is expected that the nano-sized materials may have stronger immune activation in fish than the ordinary size. Therefore, the current study was conducted to evaluate the effect of dietary chitosan nanoparticles (CNP) on growth performance, antioxidant activity, and innate immunity of Nile tilapia, Oreochromis niloticus (L.). Fish (19.8 ±â€¯0.59 g) were fed on diets enriched with 0.0, 0.25, 0.5, 1.0, and 2.0 g CNP/kg diet for 45 days. Fish performance was significantly improved with increasing CNP levels over the control diet with optimum level of 1.0 g CNP/kg diet. Antioxidant-stimulated activity was observed due to dietary CNP supplementation over the control diet in a dose-dependent manner. However, malondialdehyde level decreased significantly, whereas activities of catalase, superoxide dismutase, lysozyme, and respiratory burst increased significantly due to CNP supplementation in a dose-dependent manner. The current study evoked that dietary CNP showed strong immune modulatory properties and enhanced significantly the performance and health of Nile tilapia with optimum level of 1.0 g CNP/kg diet.

Effect of dietary purslane (Portulaca oleracea L.) leaves powder on growth, immunostimulation, and protection of Nile tilapia, Oreochromis niloticus against Aeromonas hydrophila infection.

A feeding trial was conducted to assess the effect of inclusion of dried powder of purslane, Portulaca oleracea, leaves in diets on growth, antioxidant, and immunological responses of Nile tilapia, Oreochromis niloticus, as well as its resistance to pathogenic bacteria, Aeromonas hydrophila. Fresh leaves of purslane were collected, dried, and mixed thoroughly with a basal fish diet at levels of 0.0 (control), 1.0, 2.0, and 3%. Fish (18.2 ± 0.4 g) fed one of the experimental diets up to apparent satiation twice a day for 7 weeks. At the end of the feeding trial, fish were intraperitoneally injected with pathogenic bacteria, A. hydrophila, and fish mortalities were observed and recorded up to 10 days post-challenge. Fish growth, weight gain, and specific growth rate were retarded significantly with increasing purslane levels in fish diets as compared to the control group. On the other hand, significant (P < 0.05) increases in plasmatic superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities accompanied with significant decreases in malondialdehyde (MDA) were observed with increasing purslane levels in fish diets. The respiratory burst (RB) activity and lysozyme value were significantly enhanced, while nitrous oxide (NO) decreased significantly with increasing purslane levels in fish diets over those of the control fish. After 10 days of the bacterial infection, no significant changes of MDA and SOD levels were observed, whereas CAT and GPX activities were significantly elevated. Additionally, RB and lysozyme activities were lower, while NO levels were elevated more than those before bacterial infection. Fish survival before and after bacterial infection was significantly affected by dietary purslane supplements, bacterial infection, and their interaction where dietary purslane improved the fish resistance to A. hydrophila infection in a dose-dependent manner. Purslane-fed fish showed highest relative percent of survival especially at an inclusion level of 3% (81.25%), whereas the control fish group showed lowest relative percent of survival (0.0%).

Fish response to hypoxia stress: growth, physiological, and immunological biomarkers.

Water quality encompasses the water physical, biological, and chemical parameters. It generally affects the fish growth and welfare. Thus, the success of a commercial aquaculture project depends on supplying the optimum water quality for prompt fish growth at the minimum cost of resources. Although the aquaculture environment is a complicated system, depending on various water quality variables, only less of them have a critical role. One of these vital parameters is dissolved oxygen (DO) level, which requires continuous oversight in aquaculture systems. In addition, the processes of natural stream refinement require suitable DO levels in order to extend for aerobic life forms. The depletion of DO concentration (called hypoxia) in pond water causes great stress on fish where DO levels that remain below 1-2 mg/L for a few hours can adversely affect fish growth resulting in fish death. Furthermore, hypoxia has substantial effects on fish physiological and immune responses, making them more susceptible to diseases. Therefore, to avoid disease outbreak in modern aquaculture production systems where fish are intensified and more crowded, increasing attention should be taken into account on DO levels.