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This study provides a comprehensive summary of the findings regarding the application and diagnostic efficacy of droplet digital PCR (ddPCR) in detecting viral and bacterial pathogens in aquaculture. Utilizing a systematic search of four databases up to 6 November 2023, we identified studies where ddPCR was deployed for pathogen detection in aquaculture settings, adhering to Preferred Reporting Items for Systematic Reviews and Meta-analysis of Diagnostic Test Accuracy guidelines. From the collected data, 16 studies retrieved, seven were included in a meta-analysis, encompassing 1121 biological samples from various fish species. The detection limits reported ranged markedly from 0.07 to 34 copies/μL. A direct comparison of the diagnostic performance between ddPCR with quantitative PCR (qPCR) proved challenging due to limited data, thus only a pooled sensitivity analysis was feasible. The results showed a pooled sensitivity of 0.750 (95% confidence interval [CI]: 0.487-0.944) for ddPCR, compared to 0.461 (95% CI: 0.294-0.632) for qPCR, with no statistically significant difference in sensitivity between the two methods (p = .5884). Notably, significant heterogeneity was observed among the studies (I 2  = 93%-97%, p < .01), with the year of publication significantly influencing this heterogeneity (p < .001), but not the country of origin (p = .49). No publication bias was detected, and the studies generally exhibited a low risk of bias according to QUADAS-C criteria. While ddPCR and qPCR showed comparable sensitivities in pathogen detection, ddPCR's capability to precisely quantify pathogens without the need for standard curves highlights its potential utility. This characteristic could significantly enhance the accuracy and reliability of pathogen detection in aquaculture., (© 2024 John Wiley & Sons Ltd.)

Oxygen nanobubble (NB-O 2 ) technology has been introduced to the aquaculture industry in recent years. This treatment usually results in a tremendously high level of dissolved oxygen (DO) in the water. However, little is known about the possible negative effects of hyperoxia due to NB-O 2 treatment (hyper-NB-O 2 ) on farmed fish. Here, we investigated i) the effect of short-term hyper-NB-O 2 exposure (single treatment) on the innate immunity in Nile tilapia, Oreochromis niloticus, and ii) the effect of long-term hyper-NB-O 2 exposure (26-day treatments) on survival, growth performance, gill histology, and gut microbiome in Nile tilapia. A single treatment with NB-O 2 for 10 min in 50 L of water resulted in 24.2 ± 0.04 mg/L DO (approximately 2-3 × 10 7 nanoscale oxygen bubbles/mL). This treatment did not result in differences in expression of several immune-related genes (e.g., TNF-α, LYZ and HPS70) in various tissues (e.g., gill, head kidney, and spleen) compared to the non-treated control. Over a 26-day period of exposure, no significant differences were observed in survival and growth performance of the fish, but minor histological changes were occasionally noted on the gills. Analysis of the gut microbiome revealed a significant increase in the genera Bosea, Exiguobacterium, Hyphomicrobium, and Singulisphaera in the group receiving NB-O 2 . Moreover, no signs of "gas bubble disease" were observed in the fish throughout the duration of the experiment. Overall, these results suggest that both short- and long-term hyper-NB-O 2 exposure appears to be benign and has no obvious adverse effects on fish., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This study aimed to investigate the effects of mango peel powder (MGPP) on growth, innate immunity, and immune-antioxidant related gene expression of Nile tilapia reared under biofloc system. Three hundred Nile tilapia (average weight 14.78 ± 0.05 g) were distributed into 15 fiber tanks (300 L per tank) assigned to five treatments in triplication. Fish were fed basal diet containing different levels MGPP as follows: 0 (MGPP0: control), 6.25 (MGPP 6.25), 12.5 (MGPP 12.25), 25 (MGPP 25), and 50 (MGPP 50) g kg -1 diet for 8 weeks. Specific growth rate (SGR), weight gain (WG), final weight (FW), feed conversion ratio (FCR), skin mucus of lysozyme (SMLA), and peroxidase activities (SMPA), serum of lysozyme (SL) and peroxidase (SP) were measured every for weeks; while immune-antioxidant-related gene expressions were determined after 8 weeks post-feeding. The results indicated that MGPP 25 diet resulted in higher SGR, WG, FW, and FCR but no significant differences among treatments were noticed. In terms of immune responses, lysozyme and peroxidase activities in mucus and serum were significantly higher in MGPP 12.5 and MGPP 25 diets against the control. Similarly, significant up-regulation of IL-1 and IL-8 gene expressions was observed in fish fed MGPP 25 against the control. However, no significant differences in LBP, GSTa, GPX, and GSR among treatments were observed. Overall, dietary inclusion of MGPP 25 significantly enhanced immune response and immune related gene expressions but not growth performance and antioxidant gene expressions. The results implied that MGPP can be potentially used as an immunostimulants in Nile tilapia culture., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Individual and combined efficacy of chitooligosaccharides (COS) and alginic acid (AA) at 1 g, 2 g, and 3 g per kg diet was assessed on growth and disease resistance in silver carp (Hypophthalmichthys molitrix) against Edwardsiella ictaluri. Growth parameters including specific growth rate (SGR), weight gain (WG), and feed conversion rate (FCR) were significant in fish fed 2 g and 3 g kg -1 of COS or AA, and fish fed combined COS + AA at 1, 2 and 3 kg -1 diet. In all groups, the survival rate (SR) was recorded 100%, except in group fed 2 g kg -1 AA diet. All the hematological and biochemical profiles significantly increased in groups fed 2 g and 3 g kg -1 of COS, AA, and COS + AA diets. Lipase and amylase enzyme activities and superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPx) antioxidant enzyme activities were significantly increased in fish fed 2 g and 3 g kg -1 of COS, AA, and COS + AA diet. Respiratory burst (RB), lysozyme (Lyz), reactive oxygen species (ROS) activities, and immunoglobuline (Ig) level were enhanced significantly in fish fed 2 g kg -1 of COS or COS + AA and all 3 g kg -1 diets, whereas nitric acid (NO) production and serum AP activity were improved in 2 g kg -1 COS + AA and 3 g kg -1 COS or COS + AA diets. Pro-inflammatory cytokine such as IL-8 mRNA transcriptions was significant in 2 g kg -1 COS + AA diet and all 3 g kg -1 diet. The IL-10 anti-inflammatory cytokine mRNA transcriptions were significant in 3 g kg -1 COS or COS + AA diets. This study was confirmed that H. molitrix fed with 3 g kg -1 COS or COS + AA diets were better activity when compared to other diet., Competing Interests: Declaration of competing interest All authors declare that they have no conflict of interest., (Crown Copyright © 2022. Published by Elsevier Ltd. All rights reserved.)

This study investigated the effects of dietary supplementation with anthocyanin extracted from black rice bran (AR) on the growth rate, immunological response, and expression of immune and antioxidant genes in Nile tilapia raised in an indoor biofloc system. A total of 300 Nile tilapia fingerlings (15.14 ± 0.032 g) were maintained in 150 L tanks and acclimatized for two weeks. Five experimental AR diets (0, 1, 2, 4, and 8 g kg -1 ) with various anthocyanin doses were used to feed the fish. We observed that the growth and feed utilization of fish fed with different dietary AR levels increased significantly after eight weeks (p < 0.05). In addition, the serum immunity of fish fed AR diets was much greater than that of those fed non-AR diets (p < 0.05). However, there were little or no difference in between fish fed AR enriched diets and the control AR-free diet (p > 0.05). After eight weeks, fish fed AR-supplemented diets had significantly higher mRNA transcript levels in immune (interleukin [IL]-1, IL-8, and liposaccharide-binding protein [LBP]) and antioxidant (glutathione transferase-alpha [GST-α] and glutathione reductase [GSR]) genes compared to control fish fed the AR-free diet, with the greatest enhancement of mRNA transcript levels (in the case of IL-8 by up to about 5.8-fold) in the 4 g kg -1  AR diet. These findings suggest that dietary inclusion of AR extract from black rice bran at 4-8 g kg -1 could function as a herbal immunostimulant to enhance growth performance, feed consumption, and immunity in Nile tilapia., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The current perspective is a pioneer to assess the efficacy of Salvia officinalis leave powder (SOLP) on growth, intestinal enzymes, physiological and antioxidant status, immunological response, and gene expression of Common carp (Cyprinus carpio). We also looked into fish resistance after being challenged with Aeromonas sobria, a pathogenic zoonotic bacteria. Fish (N = 120) were fed four different experimental diets in triplicate for 8 weeks. The control diet (SOLP0 - without SOLP); meanwhile, the other three diets included SOLP of 2, 4, and 8 g kg -1 concentrations (SOLP2, SOLP4, and SOLP8), respectively. Findings demonstrated that fish fed SOLP4 and SOLP8 diets had better growth performance and improved digestion by noticeable enhancing lipase and amylase enzymes activity than other groups. Additionally, the antioxidant (superoxide dismutase and glutathione peroxidase) and immune activities (immunoglobulin M, nitric oxide, and antiprotease) clarified a significant increase (p < 0.05) in SOLP4 and SOLP8 groups. Enriched diets with SOLP4 and SOLP8 exhibited better expression of splenic genes (IL-1β, IL-6, IL-10, TLR-2, and SOD), intestinal genes (Slc26a6) and (PepT1 or Slc15a1), and muscular genes (IGF-1 and SOD), while MSTN was down-regulated. After 8 weeks of the experimental trial, C. carpio challenged by A. sobria exhibited the highest cumulative mortality (66.67%), while SOLP8-dietary intervention showed the best results in enhancing the fish resistance against A. sobria by lessening mortalities to 13.33% followed by SOLP4 diet (20%). The outcomes indicate that the expression of splenic, muscular, and intestinal genes confirm the efficacy of SOLP on enhancing growth, digestion, and immune-antioxidant status, and recommend the potential use of SOLP especially at 4 g kg -1 level as a valuable natural economic diet additive in C. carpio culture for sustaining aquaculture., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

This study aimed to evaluate the effects of rambutan peel powder (RP) on growth, skin mucosal and serum immunities, and immune-related gene expression of striped catfish (Pangasianodon hypophthalmus) reared in a biofloc system. Three hundred fingerlings (17.14 ± 0.12 g fish -1 ) were randomly selected and assigned to five treatments corresponding to five diets: 0 g kg -1 (control - RP0); 10 g kg -1  (RP10); 20 g kg -1  (RP20); 40 g kg -1  (RP40), and 80 g kg -1  (RP80) for 8 weeks. At weeks 4 and 8 post-feeding, growth, skin mucus, and serum immunity parameters were determined, whereas immune-related gene expressions were performed at the end of the feeding trial. Based on the results, skin mucus lysozyme (SML) and skin mucus peroxidase (SMP) were significantly higher in fish fed the RP diets compared to the control diet (P < 0.05). The highest SML and SMP levels were observed in fish fed RP40 diet, followed by RP20, RP80, RP10, and RP0. Fish-fed RP diets had higher serum lysozyme and serum peroxidase activities, with the highest value found in the RP40 diet (P < 0.05), followed by RP20, RP80, and RP10. Similarly, immune-related gene expressions (IFN2a, IFN2b, and MHCII) in the liver were significantly up-regulated in fish fed RP40. Up-regulation (P < 0.05) of IL-1, IFN2a, IFN2b, and MHCII genes was also observed in fish intestines, with the highest values observed in fish fed RP40 diet, followed by RP10, RP20, RP80, and RP0. Fish-fed diet RP diets also showed enhanced growth and FCR compared to the control, with the highest values observed in fish fed diet RP40. However, no significant differences in survival rates were found among diets. In conclusion, dietary inclusion of RP at 40 g kg -1 resulted in better growth performance, immune response, and immune related gene expressions of striped catfish (Pangasianodon hypophthalmus)., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The current study addressed to investigate the effect of lycopene (LYC) on blood physiology, digestive-antioxidant enzyme activity, specific-nonspecific immune response, and inflammatory gene transcriptional regulation (cytokines, heat shock proteins, vitellogenins) in spotted snakehead (Channa punctata) against Pseudomonas aeruginosa. In unchallenged and challenged fish treated with 200 mg LYC enriched diet the growth performance and digestive-antioxidant enzymes increased after 30 days, whereas with inclusion of 100 or 400 mg LYC in the diets, the increase manifested on or after 45 days. No mortality in fish treated with any LYC diet against P. aeruginosa was revealed. In the unchallenged and challenged fish the phagocytic (PC) activity in head kidney (HK) and spleen were significantly enhanced when fed the control diet or other LYC diets, whereas the respiratory burst (RB) activity and nitric oxide (NO) production significantly increased when fed the 200 mg diet for 45 and 60 days. Similarly, the lysozyme (Lyz) activity in the HK and spleen, and total Ig content in serum were significantly higher in both groups fed the 200 mg LYC diet for 15, 45, and 60 days. Heat shock protein (Hsp 70) was significantly improved in the uninfected group fed the 200 mg LYC diet for 45 and 60 days, but Hsp27 did not significantly change among the experimental groups at any time points. TNF-α and IL-6 mRNA pro-inflammatory cytokine expression significantly increased in both groups fed the 200 mg LYC diet after 45 and 60 days, while the IL-12 mRNA expression was moderate in both groups fed the same diet for 60 days. The IL-10 did not significant mRNA expression between groups at any sampling. The iNOS and NF-κB mRNA expression was pointedly high in both groups fed the 200 mg LYC diet on day 45 and 60. Vitellogenin A (VgA) mRNA was significantly higher in the uninfected fish fed the 100 and 200 mg LYC diets for 45 and 60 days, but VgB did not reveal significant difference between the treatment groups at any time points. The present results suggest that supplementation of LYC at 200 mg significantly modulate the blood physiology, digestive-antioxidant enzymes, specific-nonspecific immune parameters, and cytokines, Hsp, and vitellogenins in spotted snakehead against P. aeruginosa., (Crown Copyright © 2021. Published by Elsevier Ltd. All rights reserved.)

The study was executed to find out the potential effects spent coffee ground (SCG) on Nile tilapia's skin mucosal and serum immunities, disease prevention, and growth rate reared in a biofloc system. Nile tilapia fingerlings (average weight 15.25 ± 0.07 g) were disseminated into 15 aquaria (150 L tank -1 ) at a density of 20 fish per aquarium and treated five diets: SCG1 (control), SCG2 (10 g kg -1 ), SCG3 (20 g kg -1 ), SCG4 (40 g kg -1 ), and SCG5 (80 g kg -1 ) for eight weeks. A Completely Randomized Design (CRD) with three replications was applied. Growth rate, skin mucus, and serum immunities were quantified every 4 weeks; whereas the challenge study was conducted at the termination of the feeding trial. The outputs indicated that dietary incorporation of SCG give rise to the enhancement of SGR and FCR in comparison with the control, with best levels noted in fish fed SCG2 diet. Similarly, significant enhancements in skin mucosal and serum immunities were revealed in fish treated SCG2 over the control and other SCG diets. Likewise, higher survival rates against Streptococcus agalactiae were displayed in fish fed SCG, with the maximum level displayed in the fish treated SCG2. In conclusion, dietary supplementation of SCG2 (10 g kg -1 ) can be potential used as immunostimulants in tilapia aquaculture., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Effect of bamboo vinegar powder (BVP) on growth, immunity, disease resistance, and immune-related gene expressions in juvenile Asian sea bass (barramundi), Lates calcarifer against Vibrio anguillarum was investigated. V. anguillarum infected fish fed by 2g BVP kg -1 diet exhibited significant growth, albumin (AB), serum total protein (TP), and globulin (GB) levels. Similarly, enhanced lysozyme (LZM), phagocytic (PC), anti-protease (AP), respiratory burst (RB), bactericidal (BC) activities and total immunoglobulin (Ig) levels were observed in fish fed 2g BVP kg -1 . In addition, use of 2g BVP kg -1 in diet modulated several immune related gene expressions against Vibrio disease. This study exhibited a positive effect of dietary 2g BVP kg -1 diet on hemato-biochemical and immunological responses, which enhance the immunocompetence and health status of L. calcarifer against V. anguillarum infection., (Crown Copyright © 2021. Published by Elsevier Ltd. All rights reserved.)

Effect of dietary with 100, 200, and 300 mg kg -1 glycyrrhizic acid (GA) on growth enhancer, blood physiology, digestive-antioxidant enzyme ability, innate-adaptive defense, and inflammatory cytokines induction was studied in silver carp, Hypophthalmichthys molitrix against vibriosis caused by Vibrio alginolyticus. Significant weight gain (WG), specific growth rate (SGR), and 100% survival rate (SR) was attained non-infected health (NiH) fish fed in control or all GA diets on 30, 45, and 60 days. Both NiH and V. alginolyticus challenged (VaC) fish treated with 200 mg GA diet significantly (P < 0.05) exhibited an enhancement in leucocytes value on 30, 45, and 60 days. Albumin (AB) or total proteins (TP) levels were significantly (P < 0.05) better in both groups fed 200 GA on 45 and 60 days. Malondialdehyde (MDA) and superoxide dismutase (SOD) activities were also substantial (P < 0.05) in both groups fed 200 mg GA on days 30, 45, and 60; whereas glutathione peroxidase (GPx) and catalase (CAT) activities were significantly (P < 0.05) better in both groups received 200 mg GA on days 45 and 60. Phagocytic (PC) and lysozyme (Lyz) activities significantly enhanced in both groups fed 200 or 300 mg GA on 45 and 60 days. Respiratory burst (RB), reactive oxygen species (ROS) and immunoglobulin (Ig) production significantly (P < 0.05) increased in both groups administered 200 or 300 mg GA. Growth hormone (GH) mRNA was up regulated in 200 mg GA trial on 45 days and in 200 or 300 mg GA treatments on 60 days. The IL-8 cytokine mRNA expression was up-regulated in both groups 200 and 300 mg GA on days 45 and 60, whereas TNF-α mRNA expression was increased in 200 mg GA. In addition, IL-10 cytokine mRNA expression was up regulated in 200 mg GA on 45 days whereas it was increased in both 200 mg and 300 mg GA trial on 60 days. The present study revealed that feeding fish 200 mg GA per kg diet demonstrated a better growth, digestive-antioxidant activity, innate-adaptive defense, and inflammatory cytokines induction than lower or higher dosage of GA in H. molitrix against V. alginolyticus., (Crown Copyright © 2021. Published by Elsevier Ltd. All rights reserved.)

The effect of cinnamaldehyde (CM) enriched diet on immunity and cytokine gene expression in Channa striatus against Aphanomyces invadans is reported. C. striatus was uniformly divided into eight groups (n = 25 fish each) and fed with formulated diets with 0, 5, 10, and 15 mg kg -1  CM enriched diet. In healthy and infected groups fed with 5 mg kg -1 diet the leukocytes count increased significantly after 4th week; with 10 mg kg -1  CM diet the increase manifested after 6th week, but with 15 mg kg -1 not even after 8th week. In both groups, 5 mg kg -1  CM diet resulted in a significant increase in the serum total protein, albumin, and globulin levels after 4th week, whereas with other diets this effect was observed only after 6th week. Similarly, with any enriched diet the lysozyme activity increased significantly, but with 15 mg kg -1  CM diet only after 6th week. In both groups the complement activity and lymphocyte production increased significantly when fed with 5 mg kg -1  CM diet after 4th week while with other enriched diets only after 6th week. The phagocytic activity increased significantly in both groups fed with 5 mg kg -1  CM diet after 6th week, whereas the SOD activity increased after 4th week. The IgM production increased significantly in both groups fed with 5 mg kg -1  CM diet after 2nd week, while with 5 and 10 mg kg -1  CM diet after 4th week. In both groups, the expression of CXCR3α was significant on 4th week when fed with 10 mg kg -1  CM diet, while in the healthy group fed with 15 mg kg -1  CM diet the expression manifested earlier than 4th week. However, when fed with 10 and 15 mg kg -1  CM diets the increase was observed on 6th week; whereas, the expression of MHC-I reached the maximum on 6th week with any enriched diet. The results indicate that in C. striatus the innate immunity and expression of cytokine and immune related genes were significantly modulated when fed with 5 mg kg -1  CM diet on 4th week against A. invadans., (Crown Copyright © 2021. Published by Elsevier Ltd. All rights reserved.)

This study reports the effect of ulvan enriched diet on the influence of growth, changes in hemato-biochemical indices, improvement of antioxidant system, enhancement of innate-adaptive immunity and modification of immuno-antioxidant genes expression in Labeo rohita against Flavobacterium columnaris. The weight gain (WG) was significantly high (P > 0.05) in unchallenged normal and challenged fish fed with diets enriched with 25 and 50 mg kg -1 ulvan; the FCR was better (P > 0.05) when fed with 50 mg kg -1 enriched diet. In normal fish fed with or without ulvan supplementation was noted 100% survival rate (SR). In both groups, the red blood cell (RBC) and while blood cell (WBC) counts increased significantly (P > 0.05) when fed with 50 mg kg -1 ulvan diet whereas the hemoglobin (Hb) level increased significantly on being fed with 25 and 50 mg kg -1 ulvan diets. The SOD activity was enhanced significantly in both groups fed with any dose of ulvan diets whereas the MDA and GPx activity increased only with 25 and 50 mg kg -1 ulvan diets. The phagocytic (PC) activity significantly increased with any enriched diet and control diet groups while the respiratory burst (RB) activity increased only with 50 mg kg -1 ulvan diet. The alternate complement pathway (ACP), activity of lysozyme (Lyz), and immunoglobuline M (IgM) were better in both groups fed with 50 mg kg -1 ulvan diet. The SOD and GPx antioxidant gene expression were significantly high in both groups fed with any ulvan diet while the Nrf2 gene expression was high with 50 mg kg -1 ulvan diet. The IL-1β, TNFα, hepcidin, Lyz, and IgM cytokines or proteins mRNA expression were significant in both groups fed with all ulvan supplement diet whereas the β-2M expression was significant only with 50 mg kg -1 ulvan diet. The present research indicates that both L. rohita groups fed with 50 mg kg -1 ulvan diet significantly improved growth, antioxidant system, immune defense system, and immuno-antioxidant related gene expression against F. columnaris., (Crown Copyright © 2021. Published by Elsevier Ltd. All rights reserved.)

The present study aimed to investigate the potential effects of dietary marjoram extract on growth performance, health, and disease resistance in common carp, Cyprinus carpio. To this purpose, the fish were assigned into four treatments and fed by diets supplemented with 0 (control), 100, 200, and 400 mg marjoram extract kg -1 over eight weeks and then challenged with Aeromonas hydrophila. According to the results, 200 mg kg -1 dietary marjoram extract inclusion showed the highest final weight, weight gain, and specific growth rate, and lowest feed conversion ratio (FCR). White blood cell number, Red blood cells, hematocrit, hemoglobin, mean corpuscular volume and mean corpuscular hemoglobin were markedly increased particularly at 200 mg kg -1 treatment. Marjoram extract significantly increased plasma superoxide dismutase activity and decreased malondialdehyde level compared to the control treatment. Plasma complement and lysozyme activities and total immunoglobulin levels, mucosal complement, lysozyme and alkaline phosphatase activities and immunoglobulin levels were significantly increased compared to the control group. The lowest post-challenge survival rate was observed in the control treatment, whereas the highest value was related to the 200 mg kg -1 marjoram treatment. In conclusion, the present study demonstrated that, marjoram extract is a suitable feed supplements for common carp, as it stimulates the fish growth, antioxidant, and immune systems, which suppress the fish mortality during Aeromonas septicemia. According to the results, 200 mg marjoram extract kg -1 is recommended for carp feed formulation., (Copyright © 2020. Published by Elsevier Ltd.)

The effect of diet enriched with dried lemon (Citrus limon) peel was fed to Labeo rohita at three different levels (0, 1, 2.5, and 5 g kg -1 ) for a period of 60 days; the impact of the diet on the hematology, antioxidant activity and immunological reaction and gene expression against Aeromonas sorbia is reported. In both un-challenged and challenged groups treated with 2.5 g and 5 g kg -1 dried lemon peel diets, the enhanced significant changes are: the weight gain and specific growth rate, white blood cell and total protein content, the antioxidants: superoxide dismutase, catalase, glutathione peroxidase, and glutathione activities, the respiratory burst, alternative complement pathway, complement C3, and total immunoglobulin M levels. Similarly, the heat shock protein-70 and -90, superoxide dismutase, glutathione peroxidase, glutathione, interleukin-1β and -8, tumor necrosis factor alpha, inducible nitric oxide synthase, transforming growth factor beta, and immunoglobulin M were up-regulated significantly. Any dried lemon peel enriched diet increased the phagocytic and lysozyme activities significantly in both groups. In the un-challenged group treated with 0 g kg -1 diet or in both groups treated with 2.5 g kg -1 diet the SR was 100%. These results indicate that in both un-challenged and challenged-treated groups the 2.5 and 5 g kg -1 dried lemon peel enriched diets positively modulate growth rate, physiology, and antioxidant status, innate-adaptive immune response as well as antioxidant and immune related gene expression in L. rohita against A. sorbia., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A feeding trial was carried out to examine the effects of adding chestnut (Castanea sativa) polyphenols (CSP) on the growth, skin mucus and serum immune parameters of Nile tilapia (Oreochromis niloticus). Five experimental diets with inclusion levels of 0, 1, 2, 4, and 8 g kg -1 of CSP were fed to Nile tilapia fingerlings (12.77 ± 0.17 g fish -1 ) during an eight-week trial. Fish were analyzed on the fourth and eighth week to determine the influences of CSP on growth, skin mucus, and serum immune parameters. Challenging test versus Streptococcus agalactiae was evaluated at the end of the trial. Fish fed with CSP enriched diets displayed a significant increase (P ≤ 0.05) in growth and a decline in feed conversion ratio (P ≤ 0.05). Similarly, skin mucus and serum immune parameters were significantly increased (P ≤ 0.05) in fish fed CSP with respect to the control. The effects were already evident four weeks after the CSP administration. The disease protection test displayed that the fish's survival rate was significantly higher (P < 0.05) in CSP diets over the control. The relative percentage of survival (RSP) was 62.5, 75.0, 58.3, and 37.5 in fish fed diets contained 1, 2, 4, and 8 g kg -1 CSP, respectively. The best effect on growth, immune response, and disease resistance were shown in Nile tilapia fed with a diet supplementation of 2 g kg -1 CSP., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present work, herein, studied the effects of corncob-derived xylooligosaccharides (CDXOS) and Lactobacillus plantarum CR1T5 (LP) integrated into fish diets (diet 1 (0-control), diet 2 (10 g kg -1 CDXOS), diet 3 (10 8  CFU g -1 L. plantarum CR1T5), diet 4 (10 g kg -1 CDXOS +10 8  CFU g -1 L. plantarum CR1T5)) on growth performance, innate immune parameters, and disease resistance of Nile tilapia (Oreochromis niloticus). Fingerlings, with average mean weight of 4.97 ± 0.04, were randomly distributed into 16 glass tanks (20 fish per tank) for 12 weeks. Growth performance, skin mucus, and serum immune parameters were evaluated at the conclusion of the experiment. Eight randomly selected fish were used for challenge test against Streptococcus agalactiae. The results indicated that fish fed CDXOS and LP had significantly improved final weight (FW), weight gain (WG), specific growth rate (SGR), and feed conversion ratio (FCR). However, no significant difference in survival rate was observed between specimens fed the supplemented diets and the control. Regarding skin mucus, the dietary inclusion of CDXOS and LP significantly increased lysozyme and peroxidase activities compared with the control (P < 0.05). Similarly, significant increases in serum lysozyme, peroxidase, alternative complement, phagocytosis, and respiratory burst activities were observed in the fish fed the supplemented diets. However, no significant differences were found in these parameters between fish fed CDXOS and LP diets. For the challenge test, diet 4 produced a higher relative percentage of survival (RPS) and resistance to S. agalactiae than fish from the other experimental groups (P < 0.05). The results suggested that CDXOS and L. plantarum CR1T5 are viable considerations for potential feed-additive sources.

A trial was operated to assess the potential of using Lactobacillus plantarum L-137 (L-137) and/or β-glucan (BG) in improving the resistance of Nile tilapia against Aeromonas hydrophila. Control diet and 3 diets supplemented with L-137, BG or L-137 + BG were prepared. Final body weight, specific growth rate, superoxide dismutase, and catalase showed considerably (P < .05) increased values in L-137 or L-137/BG groups, while glutathione peroxidase increased significantly (P < .05) only in L-137/BG group. Fish fed L-137 and/or BG diets showed that feed conversion ratio and malonaldehyde levels were significantly decreased (P < .05). Also, both L-137 and BG helped Nile tilapia to have high phagocytosis activity and relative expression of tumor necrosis factor-alpha (TNF-α) and interleukin 1 beta (IL-1β) and interferon-gamma (INF-γ) genes. After A. hydrophila challenge, the intestinal villi epithelium of the L-137/BG group was intact and denser than the other groups. The hepatopancreas and spleen of the control group displayed severe necrosis in hepatocytes and congestion of blood sinusoids in addition to diffuse vacuolation. Regarding the L-137, BG and L-137/BG groups, there was a moderate and normal degree of vacuolation with focal necrosis and mild to moderate degree of congestion of blood sinusoids. Red blood cells, hemoglobin, and albumin showed meaningfully (P < .05) increased values in L-137 or L-137/BG groups. TNF-α, IL-1β, and INF-γ expressions were upregulated by L-137 and/or BG. The obtained results revealed the ability of L-137 and/or BG to protect Nile tilapia from the effects of A. hydrophila infection by the motivation of the immune, antioxidative, and antiinflammation responses., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present study was investigated the dietary administration of cassic acid (CA) on growth, innate immunity, transcription profiles of estrogen and follicle-stimulating hormones as well as lysozyme enzyme determined in Clarias gariepinus against Edwardsiella tarda. The weight gain (WG), protein efficacy ratio (PER), and feed conversion ratio (FCR) were significantly improved in infected fish fed dietary administration with CA at 5 and 10 mg kg -1 diets. The survival is higher (96.7% and 98.3%) in the infected groups fed at 5 and 10 mg kg -1  CA diets. The red (RBC) and white (WBC) blood cells, hemoglobin (Hb), and packed cell volume (PCV) was found significantly high in the infected fish feeding at 5 and 10 mg kg -1  CA diets. Total protein and albumin were significantly increased with 5 and 10 mg kg -1  CA diets among weeks 1-4 while the globulin and albumin: globulin ratio increased of these diet only after week 2. The phagocytic and respiratory burst activities were enhanced statistically the infected fish fed at 10 mg kg -1  CA diet group whereas the production of superoxide anion (SOA) and nitric oxide (NO) were significantly increased at 5 and 10 mg kg -1  CA diets. The lymphocyte proliferation and myeloperoxidase (MPO) activity were significantly high the infected fish when fed at 5 and 10 mg kg -1  CA diets after 2nd week whereas the alternate complement activity (ACP), generation of reactive oxygen species (ROS), and lysozyme activity (Lyz) were observed at 5 and 10 mg kg -1  CA diets among weeks 1-4. The accumulative mortality was 10% in infected fish fed at 1 and 5 mg kg -1  CA diets whereas 15% mortality found with 10 mg kg -1  CA diet. The highest levels of estrogen receptor alpha (ERα) mRNA expression found in gonad while the highest levels of follicle stimulating hormone-beta subunit (FSH-β) mRNA expression found in testis of the infected fish given at 5 and 10 mg kg -1  CA diets. The up-regulation of chick-type lysozyme (c-Lyz) mRNA was observed at 5 and 10 mg kg -1  CA diets after 2nd week while goose-type lysozyme (g-Lyz) mRNA was up-regulation amongst weeks 1-4 of these diets. The present study suggested that E. tarda challenged fish after feeding with 5 and 10 mg kg -1  CA diets did not affect growth and hemato-biochemical parameter, but it enhanced nonspecific immune system and improving ERα, FSH-β, c-Lyz, and g-Lyz mRNA expression in C. gariepinus against E. tarda., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present study aimed to assess the possible effects of Assam tea (Camellia sinensis) extract (ATE) on growth performances, immune responses, and disease resistance of Nile tilapia, Oreochromis niloticus against Streptococcus agalactiae. Five levels of ATE were supplemented into the based diet at 0, 1, 2, 4, and 8 g kg -1 feed of Nile tilapia fingerlings (10.9 ± 0.04 g initial weight) in triplicate. After four and eight weeks of feeding, fish were sampled to determine the effects of the tea supplements upon their growth performance, as well as serum and mucosal immune responses. A disease challenge using S. agalactiae was conducted at the end of the feeding trial. Fish fed ATE revealed significantly improved serum lysozyme, peroxidase, alternative complement (ACH50), phagocytosis, and respiratory burst activities compared to the basal control fed fish (P < 0.05). The mucus lysozyme and peroxidase activities were ameliorated through ATE supplementation in the tilapia diets. Supplementation of ATE significantly (P < 0.05) enhanced final body weight, weight gain, and specific growth rate; while a decreased feed conversion ratio was revealed at 2 g kg -1 inclusion level, after four and eight weeks. Challenge test showed that the relative percent survival (RSP) of fish in each treatment was 33.33%, 60.00%, 83.33%, 76.68%, and 66.68% in groups fed 0, 1, 2, 4, and 8 g kg -1 , respectively. In summary, diets supplemented with ATE especially at 2 g kg -1 increased the humoral and mucosal immunity, enhanced growth performance, and offered higher resistance against S. agalactiae infection in Nile tilapia., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

This investigation was aimed to determine the efficacy of coriander seed extract (Coriandrum sativum) on physiological responses, immunity and disease resistance of rainbow trout, Oncorhynchus mykiss for eight weeks. A total number of six hundred rainbow trout (62 ± 0.81 g) were divided into four feeding groups including 0 (control), 0.5%, 1% and 2% of coriander seed extract (CSE). In the present study, rainbow trout fed with 2% of CSE showed significantly higher values of specific growth rate (SGR), final weight (FW) and condition factor (CF) in comparison with control group after eight weeks (P < 0.05). Regarding hematological indices results, the 2% dosage of CSE showed the highest amount of hematocrit and hemoglobin compared to control group (P < 0.05). In addition, significant improvement of lysozyme and alternative complement activity, were observed in 2% of CSE treatment (P < 0.05). After eight weeks post-feeding, 30 fish from each treatment were challenged with Yersinia ruckeri for 14 days. The findings presented that fish fed with CES, especially 2% of CSE inclusion, improved survival rate of rainbow trout against Y. ruckeri; however, there were no significant differences among the fish in control and treatment groups at the end of the eight weeks feeding with coriander seed extract. The present study demonstrated, dietary incorporation of coriander extract can improve growth factors, immunological indices and resistance of rainbow trout (O. mykiss) against Y. ruckeri infection., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The present study evaluates the effects of dietary kefir and low molecular weight sodium alginate (LWMSA) (singular or combined) on non-specific immune response, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus). Fish with average weight of 18.60 ± 0.04 g were supplied and randomly stocked in sixteen glass tanks (150 L) at density of 20 fish per tank. Fish were fed experimental diets as follows: 0 g kg -1 LMWSA (Control, Diet 1), 10 g kg -1 LMWSA (Diet 2), 40 g kg -1 kefir (Diet 3), and 10 g kg -1 LMWSA + 40 g kg -1 kefir (Diet 4) for 50 days. At the end of the feeding trial, serum lysozyme (SL), phagocytosis (PI), respiratory burst (RB), and alternative complement (ACH50) activities as well as growth performance were measured. Singular and combined administration of kefir and low molecular weight sodium alginate (LMWSA) significantly increased serum SL, PI, RB, and ACH50 activities compared control group (P < 0.05); the highest innate immune responses were observed in fish fed combinational diet (kefir + LMWSA) (P < 0.05). The results of experimental challenge revealed significantly higher resistance against Streptococcus agalactiae in fish fed supplemented diets and the highest post challenge survival rate was observed in synbiotic diet (P < 0.05). Similar results obtained in case of growth parameters. Feeding on supplemented diet significantly improved SGR and FCR and the highest growth parameters was observed in fish fed synbiotic diet (P < 0.05). These finding revealed that combined administration of dietary kefir and LMWSA can be considered for improving immune response, disease resistance and growth performance of Nile tilapia., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Present study was carried out to evaluate the effects of low molecular weight sodium alginate (LMWSA) as potential prebiotic source on growth performance, innate immunity and disease resistance of tilapia, Oreochromis niloticus. Three hundred twenty fish were divided into four treatments and fed following diets 0 (T1- Control), 10 (T2), 20 (T3) and 30 (T4) g kg(-1) LMWSA for period of 60 days. A Completely Randomized Design with four replications was applied. At the end of experiment, fish in each replication were weighed and specific growth rate (SGR) and feed conversion ratio (FCR) were calculated. Five randomly selected fish were used for innate immune response measurement. Another ten fish were randomly selected for challenge test against Streptococcus agalactiae for a period of 18 days. The lysozyme, complement, phagocytosis, and respiratory burst activities were detected after 60 days of feeding trial and after challenge test. The results indicated that fish fed diet 10 g kg(-1) LMWSA significantly improved SGR and FCR after 60 days of feeding trial. The lysozyme, phagocytosis, respiratory burst, and complement activities were significantly higher in fish fed LMWSA diets compared to control. Fish fed 10 g kg(-1) LMWSA had greatest values compared to fish fed 20 and 30 g kg(-1) LMWSA. The survival rate of O. niloticus was significantly improved in fish fed LMWSA diets after challenge with S. agalactiae for 18 days. However, no significant difference in survival rate was observed among LMWSA supplemented diets. It is indicated that fish fed 10, 20 and 30 g kg(-1) LMWSA diets can stimulate growth performance, innate immunity and disease resistance in tilapia against S. agalactiae., (Copyright © 2016 Elsevier Ltd. All rights reserved.)