Synergetic response on herbal and probiotic applications: a review.

Herbs and their by-products are important traditional medicines and food supplements; they provide numerous beneficial effects for animals. Consequently, probiotics are living cell organisms, nontoxic, and friendly microbes. Probiotics have numerous beneficial activities such as inhibition of pathogens, enhancement of the immune system, growth, disease resistance, improving water quality, reducing toxic effects, synthesis of vitamins, prevention of cancer, reduction of irritable bowel syndrome, and more positive responses in animals. Herbal and probiotic combinations have more active responses and produce new substances to enhance beneficial responses in animals. Herbal and probiotic mixture report is still limited applications for animals. However, the mechanisms by which they interact with the immune system and gut microbiota in animals are largely unclear. This review provides some information on the effect of herbal and probiotic blend on animals. This review discusses current research advancements to fulfill research gaps and promote effective and healthy animal production.

Toxic and bioaccumulative effects of zinc nanoparticle exposure to goldfish, Carassius auratus (Linnaeus, 1758).

The widespread use of produced metal oxide nanoparticles (NPs) has increased major concerns about their impact on human as well as aquatic animal health. The present study shows that exposure to different concentrations of zinc oxide (ZnO) NPs led to high accumulations of Zn ions in the metabolic organs of fish (liver and gills), resulting in severe oxidative stress in Carassius auratus. The goldfish (C. auratus) was chosen as an aquatic species for the evaluation of the potential toxicity of aqueous ZnO-NPs (Treatments of hemoglobin and neutrophils (0, 0.5, 1, and 1.5 mg L- 1) following 14 days of exposure. A range of histological and hematological factors were examined. Exposure to the NPs produced significant reduction of red blood cell and white blood cell counts, hematocrit) were found to produce no significant differences in lymphocyte, monocyte, and eosinophil counts; as well as the mean corpuscular hemoglobin concentrations index (P > 0.05). Moreover, the results revealed significant alterations in serum biochemical parameters, hepatic enzyme levels, and immune and antioxidant responses; except for total protein and superoxide dismutase (SOD) of C. auratus exposed to ZnO-NPs, particularly at the 1 and 1.5 mg L- 1 concentrations. Fish exposed to 1 and 1.5 mg L-1 ZnO-NPs displayed a significant reduction in alternative complement pathway activity, lysozyme, and total protein contents of mucus compared to those in the control group. The results showed that hepatic SOD and catalase, and gill catalase activity were significantly decreased, and their malondialdehyde levels increased at 1 and 1.5 mg L-1 ZnO-NPs compared to the control group (P < 0.05). Significant accumulations of ZnO-NPs were observed in the liver, kidney, and gill tissues of fish leading to severe histopathological alterations in these organs. These results suggest that water-borne ZnO-NPs can easily accumulate in metabolic organs and lead to oxidative stress and destructive effects on the physiological features of C. auratus.

The Effects of Laurencia caspica Algae Extract on Hemato-Immunological Parameters, Antioxidant Defense, and Resistance against Streptococcus agalactiae in Nile tilapia (Oreochromis niloticus).

Natural immune stimulants are among the most effective chemicals for boosting immunity and fish welfare. This study aims to investigate the effects of red macroalgae extract (Laurencia caspica) on hematological, immunological, antioxidant, biochemical, and disease resistance against S. agalactiae in Nile tilapia for 50 days. For this purpose, fishes were assigned to four dietary treatments group in which the base meal was supplemented with 0.5%, 1%, and 2% of L. caspica extract. On days 25 and 50 of the experiment, samples were taken to investigate the hematological, immunological, biochemical, and antioxidant parameters. The white blood cells (WBCs), hemoglobin, and neutrophils significantly increased after 50 days of feeding with the L. caspica extract, but until the 25th day, no significant difference was observed among the treatments except for hemoglobin. Immunological parameters (including Immunoglobulin M [IgM] and complement 3 [C3]) were significantly higher in treated groups compared to control both 25 days and 50 days posttreatment. However, on the 25th day, no significant difference was noticed between treatments and control in the case of lysozyme activity. Alkaline phosphatase (ALP) and alanine aminotransferase (ALT) considerably increased in comparison to the control group on the 50th day, but no significant difference was observed on the 25th day. In addition, feeding with L. caspica significantly increased the antioxidant enzyme activities on the 25th day (L. caspica 1% and 2% in peroxidase [POD] and superoxide dismutase [SOD] in all groups) and 50th day (catalase [CAT], SOD and L. caspica 1% and 2% in POD) in the spleen. The survival rate of fish challenged with Streptococcus agalactiae was considerably greater than the control group. Finally, it can be concluded that L. caspica extract is an immunological stimulant that induces fish resistance to S. agalactiae.

Effect of Different Levels of Chlorogenic Acid on Growth Performance, Immunological Responses, Antioxidant Defense, and Disease Resistance of Rainbow Trout (Oncorhynchus mykiss) Juveniles.

The current study is designed to assay the efficacy of chlorogenic acid (ChA) in the diet on growth performance, digestive enzyme activity, serum immunological, biochemical, and antioxidant variables, and mucosal immune response as well as disease resistance of rainbow trout (Oncorhynchus mykiss) juveniles. Rainbow trout juveniles received diets supplemented with different inclusion levels of ChA (0 (ctrl), 200 (CA1), 400 (CA2), 600 (CA3), and 800 (CA4) mg kg-1 diet) for 60 days. According to the findings, fish from CA3 and CA4 groups demonstrated the best results considering the final weight (FW) and weight gain (WG) (P < 0.05). Also, the group that received 600 mg kg-1 ChA-supplemented diet showed the lowest feed conversion ratio (FCR) and the highest specific growth rate (SGR) compared to other groups (P < 0.05). Moreover, the minimum survival rate (SR) was only detected in the CA4 treatment (P < 0.05). Regression analysis exhibited that rainbow trout growth indices were polynomially linked to dietary chlorogenic acid concentrations. In this regard, the optimal levels of chlorogenic acid according to growth parameters (FCR and SGR) were 0.71 and 0.62 gr kg-1 diet, respectively. The results exhibited superior performance of protease and amylase activities in CA2, CA3, and CA4 groups with the maximum amount in the group receiving 600 mg kg-1 ChA-enriched diet (P < 0.05). Serum lysozyme (LYZ), immunoglobulin (Ig), and components 3 and 4 (C3 and C4) values of CA2, CA3, and CA4 groups were significantly higher than others with the highest amount in the CA3 group (P <0.05). Additionally, serum nitroblue tetrazolium (NBT) value in the CA3 and CA4 groups and myeloperoxidase (MPO) in the CA3 group were notably more than others (P < 0.05). Moreover, the lowest aspartate aminotransferase (AST), alkaline phosphatase (ALP), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) and the highest total protein (TP) and globulin (GLO) values were observed in CA3 treatment (P < 0.05). CA2 and CA3 groups demonstrated increased serum catalase (CAT) and decreased malondialdehyde (MDA) values compared to the control while the highest CAT and lowest MDA values were observed in CA3 treatment (P < 0.05). Considering mucus immunity, the significantly maximum LYZ and protease values were demonstrated in CA2 and CA3 groups, and the highest ALP, Ig, and esterase values were demonstrated in the CA3 group. In comparison with the control, the mortality rates of the groups that received the ChA diets were remarkably (P < 0.05) lower postchallenge with Y. ruckeri, and the highest survival and relative percentage of survival (RPS) (P < 0.05) belonged to the CA3 group. Results obtained from the current study suggested ChA as a functional dietary additive to raise growth parameters, immune indices, antioxidant capacity, and resistance to disease in rainbow trout.

Effects of Dietary Pediococcus acidilactici and Isomaltooligosaccharide on Growth Performance, Immunity, and Antioxidant Defense in Juvenile Common Carp.

The aim of this study was to investigate the synbiotic effects of Pediococcus acidilactici (PA) and isomaltooligosaccharide (IMO) on the performance of juvenile common carp (Cyprinus carpio). A total of 360 fish (17.22 ± 0.19 g) were randomly divided into six groups with three replicates of 20 fish each. The trial continued for 8 weeks. The control group was fed only basal diet; PA was fed basal diet supplemented with 1 g/kg (1010 CFU/kg) PA, IMO5 (5 g/kg IMO), IMO10 (10 g/kg IMO), PA-IMO5 (1 g/kg PA and 5 g/kg IMO), and PA-IMO10 (1 g/kg PA and 10 g/kg IMO). The results indicated that the diet containing 1 g/kg PA and 5 g/kg IMO significantly increased the fish growth performance and decreased the feed conversion ratio (p < 0.05). Overall, blood biochemical parameters, serum (lysozyme, complements C3 and C4) and mucosal (protein, total immunoglobulin, and lysozyme) immune responses, and antioxidant defense of fish also improved in the PA-IMO5 group (p < 0.05). Therefore, a combination of 1 g/kg (1010 CFU/kg) PA and 5 g/kg IMO can be recommended as a beneficial synbiotic additive and immunostimulant in juvenile common carp.

The Study of Caspian Roach (Rutilus caspicus) Fry Health Fed with Phytobiotic-Supplemented and Salinity Stress Resistance with Emphasis on Gill Tissue Pathology.

Herbal treatment augments immune and antioxidant responses and suppresses stress in fish. Ginger (Zingiber officinale) is a popular plant with medicinal uses because of its immunostimulant, antimicrobial, and antioxidant characteristics. This study aimed to investigate the effects of ginger (Z. officinale) on growth, digestive enzymes activity, antioxidant and immune response, and salinity stress resistance of Caspian roach (Rutilus caspicus). Fish (0.98 ± 0.09 g) were divided into four treatment-fed diets containing 0 (control, Z0), 10 (Z10), 20 (Z20), and 30 (Z30) g/kg ginger powder for 56 days and then subjected to 2 g/L salinity stress for 48 hr. The highest final weight of specific growth rate (SGR), superoxide dismutase (SOD), and catalase activity and the lowest final weight of feed conversion ratio (FCR) and malondialdehyde (MDA) were observed in fish-fed diets containing 10 and 20 g/kg ginger inclusion. Intestinal protease activity significantly increased in Z10 treatment, and the highest amylase and lipase activities were related to control, including 10 g/kg ginger in the diet compared to the control group had a significant effect on immune indices such as immunoglobulin M (IgM) and ACH50 (p < 0.05). The highest lysozyme was obtained in Z20 treatment, which had a significant difference in the control (p < 0.05). Cortisol and glucose levels were significantly lower in ginger treatments than the control before and/or after salinity stress. Histopathologic results showed that hyperplasia, edema, expansion of secondary lamella, epithelial cells, and necrosis of gills were most common lesions. However, the results of this study demonstrate that using ginger powder in addition to improving of growth, it can be also effective in survival rate of Caspian roach fingerling as an endangered species exposed to salinity stress.

Quercetin Application for Common Carp (Cyprinus carpio): I. Effects on Growth Performance, Humoral Immunity, Antioxidant Status, Immune-Related Genes, and Resistance against Heat Stress.

This study was done to evaluate the effect of different quercetin levels on growth performance, immune responses, antioxidant status, serum biochemical factors, and high-temperature stress responses in common carp (Cyprinus carpio). A total number of 216 common carp with an average weight of 27.21 ± 53 g were divided into 12 tanks (four treatments × three replications) and fed 0 mg/kg quercetin (T0), 200 mg/kg quercetin (T1), 400 mg/kg quercetin (T2), and 600 mg/kg quercetin (T3) for 60 days. There were significant differences in growth performance, and the highest final body weight (FBW), weight gain (WG), specific growth rate (SGR), and feed intake (FI) were observed in T2 and T3 (P < 0.05). Different quercetin levels significantly increased complement pathway activity (ACH50) and lysozyme activity both before and after heat stress (P < 0.05). Catalase (CAT), glutathione peroxidase (GPx), and malondialdehyde (MDA) were significantly increased in fish exposed to heat stress, but fish fed with a supplemented diet with quercetin showed the lowest levels both before and after heat stress (P < 0.05). Superoxide dismutase (SOD) levels were significantly enhanced in fish fed diets supplemented with quercetin in both phases (P < 0.05). Different quercetin levels led to a significant decrease in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) before and after the challenging test (P < 0.05). Glucose and cortisol levels were significantly higher in the control group compared to the other treatments in both phases (P < 0.05). The expression of glutathione peroxidase (GPx) and lysozyme was markedly upregulated in fish fed with quercetin-supplemented diets (P < 0.05). No marked effects were observed for growth hormone (GR) and interleukin-8 (IL8) (P > 0.05). In conclusion, dietary quercetin supplementations (400-600 mg/kg quercetin) improved growth performance, immunity, and antioxidant status and increased tolerance to heat stress.

Modulation of humoral immunological and antioxidant responses and gut bacterial community and gene expression in rainbow trout, Oncorhynchus mykiss, by dietary lactic acid supplementation.

The present study was conducted to find the effects of dietary lactic acid (LA) supplementation on growth performance, hematological characteristics, humoral antioxidant and immunological factors, intestinal gene expression and bacteriological parameters in rainbow trout, Oncorhynchus mykiss. The fish (104 ± 0.90 g) were randomly distributed in twelve tanks (150 L water) as four treatments, receiving diets supplemented with 0, 5, 10, and 20 g/kg LA for 70 days. At the end of the feeding trial, no significant differences were observed in the fish growth performance and survival among the treatments. Blood/plasma red blood cell (5-20 g/kg LA), white blood cell (10 g/kg LA), hematocrit (5 and 10 g/kg LA), hemoglobin (5 and 10 g/kg LA), superoxide dismutase (10 g/kg LA), glutathione peroxidase (10 g/kg LA), total antioxidant capacity (10 g/kg LA), lysozyme (5-20 g/kg LA), and alternative complement activity (10 g/kg LA) significantly increased, as malondialdehyde (10 g/kg LA) significantly decreased in the LA-treated fish. Dietary LA supplementation induced no significant changes in the intestinal defensin gene expression, but significantly up-regulated the expression of toll-like receptor-3 (5-20 g/kg LA), Mx-9 (5-20 g/kg LA), and heat shock protein-70 (5 g/kg LA) genes; the highest expressions were observed at 5 g/kg LA supplementation. Dietary LA supplementation significantly increased total bacterial count (5 and 10 g/kg LA), Lactobacillus sp. (5-20 g/kg LA), and Bacteroides sp. (5-20 g/kg LA), but decreased Streptococcus iniae (5-20 g/kg LA) in the fish intestine; the highest Lactobacillus sp. And Bacteroides sp. Were observed in 10 and 5 g/kg LA supplementation, respectively. Overall, dietary LA supplementation, although has no significant effects on the fish growth performance, is beneficial to improve humoral antioxidant parameters, humoral and intestinal immunological parameters and intestinal microflora. According to the results, dietary 5-10 g/kg LA is recommended for the formulation of rainbow trout feed.

Dietary red macroalgae (Halopithys incurva) improved systemic an mucosal immune and antioxidant parameters and modulated related gene expression in zebrafish (Danio rerio).

In the present study, the effects of dietary Halopithys incurva, a red macroalgae species, (0.25, 0.50, 1%) on whole-body serum and skin mucus immune system, antioxidant system and expression of various genes in zebrafish were investigated. At the end of the 8-week study, total protein, total immunoglobulin and lysozyme activities in whole-body serum and skin mucus increased in fish fed H. incurva (P < 0.05). While an increase was observed in superoxide dismutase (SOD), Catalase (CAT) and glutathione peroxidase (GPx) which are antioxidant enzyme activities in whole-body serum and skin mucus, a decrease in malondialdehyde (MDA) levels was detected (P < 0.05). All of the immune-related genes examined, such as Interleukin-1 beta (IL-1ß), Tumor necrosis factor-alpha (TNF-α), Interferon-gamma (INF- γ), were upregulated by the addition of 0.5% H. incurva (P < 0.05). While SOD and GPx, which are antioxidant enzyme-related genes, were remarkably upregulated in macroalgae fed fish (P < 0.05), no change was observed in CAT gene expression (P > 0.05). Based on the results of this study, it is considered that the addition of 0.5% H. incurva to the diets of commercially farmed fish will increase their immune and antioxidant defences and may contribute to the aquaculture sector for more sustainability.

Bioactive immunostimulants as health-promoting feed additives in aquaculture: A review.

Bioactive immunostimulants could be derived from different sources like plants, animals, microbes, algae, yeast, etc. Bioactive immunostimulants are the most significant role to enhance aquatic production, as well as the cost of this method, which is effective, non-toxic, and environment-friendly. These immunostimulants are supportive to increase the immune system, growth, antioxidant, anti-inflammatory, and disease resistance of aquatic animals' health and also improve aquatic animal feed. Diseases are mainly targeted to the immune system of aquatic organisms in such a way that different processes of bioactive immunostimulants progress are considered imperative techniques for the development of aquaculture production. Communicable infections are the main problem for aquaculture, while the mortality and morbidity connected with some outbreaks significantly limit the productivity of some sectors. Aquaculture is considered the mainly developing food production sector globally. Protein insists is an important issue in human nutrition. Aquaculture has been an exercise for thousands of years, and it has now surpassed capture fisheries as the most vital source of seafood in the world. Limited study reports are available to focal point on bioactive immunostimulants in aquaculture applications. This review report provides information on the nutritional administration of bioactive immunostimulants, their types, functions, and beneficial impacts on aquatic animals' health as well as for the feed quality development in the aquaculture industry. The scope of this review combined to afford various kinds of natural derived bioactive molecules utilization and their beneficial effects in aquaculture applications.

Dietary plant pigment on blood-digestive physiology, antioxidant-immune response, and inflammatory gene transcriptional regulation in spotted snakehead (Channa punctata) infected with Pseudomonas aeruginosa.

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.

Effects of dietary rambutan (Nephelium lappaceum L.) peel powder on growth performance, immune response and immune-related gene expressions of striped catfish (Pangasianodon hypophthalmus) raised in biofloc system.

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).

Influences of spent coffee grounds on skin mucosal and serum immunities, disease resistance, and growth rate of Nile tilapia (Oreochromis niloticus) reared under biofloc system.

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.

Effects of dietary thyme essential oil and prebiotic administration on rainbow trout (Oncorhynchus mykiss) welfare and performance.

The aim of this study was to examine the combined effects of thyme essential oil (TEO) and prebiotic (Immunogen ®) on growth performance, hematological parameters, innate immunity, and oxidative status of rainbow trout, Oncorhynchus mykiss. For this purpose, the fish (11.92 ± 0.06 g) were fed (3% of biomass) experimental diets that contained 0 (Control; without Immunogen and/or TEO); 1% TEO + 0.1% Immunogen (T1); 1% TEO + 0.2% Immunogen (T2); 2% TEO + 0.1% Immunogen (T3) and 2% TEO + 0.2% Immunogen (T4) for 60 days. According to results, all experimental treatments exhibited similar final weight, weight gain, specific growth rate (SGR), and survival rate (SR), which were significantly higher than those of the control treatment. Although all treatments reduced the feed conversion ratio (FCR) compared to the control group, the lowest value was observed in T4. All experimental treatments showed a significant increase in amylase and protease activity compared to the control group. Moreover, the fish fed on T4 and T1 diets showed the highest and lowest lipase activity. Dietary TEO and Immunogen supplementations significantly increased WBC count and Hb level compared to the control group. Fish fed on control and T2 diets displayed lower hematocrit than fish fed on other experimental diets. The highest and lowest MCH index were recorded in T3 and control groups, respectively. The fish fed diets supplemented with T3 diet presented significantly higher MCV index compared to the control and T2 treatment. All experimental treatments exhibited similar AST, and ALP activities, which were significantly lower than those of the control group. Also, the lowest ALT activity was observed in T2 and T4 treatments compared to other groups. Dietary TEO and Immunogen supplementations significantly enhanced skin mucus total Ig, total protein level, and ACH50, protease, and lysozyme. All experimental treatments exhibited enhanced intestine total Ig, ACH50, and lysozyme level. Dietary thyme essential oil and Immunogen supplementations significantly enhanced liver antioxidant parameters including catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and superoxide dismutase (SOD) and decreased MDA production compared to fish fed on control diet. As a result, it can be suggested that the combination of thyme essential oil and Immunogen (specially 2% TEO + 0.2% Immunogen) is useful for enhancing the yield and well-being of farmed rainbow trout.

Effect of a diet enriched with sodium propionate on growth performance, antioxidant property, innate-adaptive immune response, and growth-related genes expression in critically endangered beluga sturgeon (Huso huso).

Organic acids are active substances required for improving the productivity and wellbeing of aquatic animals. Herein, the study investigated the effects of sodium propionate on growth performance, antioxidative and immune responses, and growth-related genes expression in beluga sturgeon (Huso huso). For eight weeks, fish fed sodium propionate at 0, 1.2, 2.5, and 5 g kg-1. The final weight, weight gain, and SGR were substantially increased while FCR decreased by dietary sodium propionate at 2.5 and 5 g kg-1 (P < 0.05). The expression of Growth hormone (GH) and insulin-like growth factor 1 (IGF-1) was markedly upregulated (P < 0.05) by dietary sodium propionate in the gills and livers of beluga. The highest mRNA level of GH and IGF-1 has been observed in fish fed a 2.5 g sodium propionate/kg diet. The red blood cells count, and hemoglobin level were meaningfully increased (P < 0.05) by 2.5 and 5 g sodium propionate/kg diet compared with 0 and 1.2 g kg-1 levels. Further, the hematocrit level was increased (P < 0.05) by a dietary 5 g sodium propionate/kg diet. The total protein level and lysozyme activity were meaningfully increased (P < 0.05) by 2.5 and 5 g sodium propionate/kg diet compared with 0 and 1.2 g kg-1 levels. The highest superoxide dismutase was observed in fish fed 2.5 g sodium propionate/kg diet. Catalase activity was significantly higher in fish fed 5 g kg-1 than 1.2 g kg-1. The glutathione peroxidase activity was markedly higher in fish fed 2.5, and 5 g kg-1 than fish fed control diet. The lowest malondialdehyde levels were observed in fish fed 1.2, and 2.5 g sodium propionate/kg diets. Moreover, the highest mucosal total protein, total immunoglobulin and lysozyme were recorded in fish fed 2.5, and 5 g sodium propionate/kg diets. The obtained results indicate that dietary sodium propionate is recommended at 2.5-5 g kg-1 to improve beluga sturgeon's growth performance, feed utilization, and wellbeing.

Influence of black rice (Oryza sativa L.) bran derived anthocyanin-extract on growth rate, immunological response, and immune-antioxidant gene expression in Nile tilapia (Oreochromis niloticus) cultivated in a biofloc system.

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.

Effect of Dietary Administration of Green Macroalgae (Ulva intestinalis) on Mucosal and Systemic Immune Parameters, Antioxidant Defence, and Related Gene Expression in Zebrafish (Danio rerio).

This study investigated the effects of adding green macroalgae gutweed (Ulva intestinalis) powder to zebrafish (Danio rerio) feed at different levels on innate immune responses, antioxidant defence, and gene expression. A total of 600 zebrafish (0.3 ± 0.08 g) were randomly allocated to 12 aquariums in four treatments with three replicates (50 fish per aquarium). Zebrafish were fed with different levels of U. intestinalis powder 0, 0.25, 0.5, and 1% for eight weeks. Whole-body extract (WBE) immune parameters including total protein level, globulin level, and lysozyme activity were evaluated and revealed statistically significant increased in all U. intestinalis supplemented groups compared to the control (P < 0.05). However, mucus immune parameters (total protein, globulin, and lysozyme) were statistically different in only 1% gutweed supplemented groups from other groups. While glutathione peroxidase (GPx) and superoxide dismutase (SOD) increased with the addition of gutweed (P < 0.05), catalase (CAT) did not change (P > 0.05). The study results showed that dietary gutweed remarkably upregulated immune-related genes such as lysozyme (Lyz) and Interleukin 1 beta (IL-1ß). Antioxidant-related genes (SOD and CAT) and growth-related genes, including growth hormone (GH) and insulin-like growth factor-I (IGF-1), were remarkably upregulated with gutweed treatment (P < 0.05). In conclusion, dietary U. intestinalis showed beneficial effects on immunity, and same effects were observed in case of antioxidant and growth related genes expression in zebrafish.

The Effects of Dietary Thyme Oil (Thymus vulgaris) Essential Oils for Common Carp (Cyprinus carpio): Growth Performance, Digestive Enzyme Activity, Antioxidant Defense, Tissue and Mucus Immune Parameters, and Resistance against Aeromonas hydrophila.

This study evaluated the effects of dietary supplementation of thyme (Thymus vulgaris) essential oil (TVO) on growth performance, digestive enzymes, biochemical parameters, hematological indices, liver enzymes, and pathogen resistance in common carp (Cyprinus carpio). Triplicate groups of fish (15.36 ± 0.10 g) were fed daily with diets supplemented with TVO at 0, 0.5, 1, and 2 percent for 60 days then challenged with Aeromonas hydrophila. The results determined that supplementation of thyme resulted in significantly higher final body weights (FBW) and lower feed conversion ratios (FCR). Furthermore, no mortality was observed in the thyme-supplemented treatments. Regression analysis showed that fish growth parameters were polynomially related to dietary TVO levels. The optimum dietary TVO level, based upon the varied growth parameters, was 1.344 to 1.436%. Digestive enzymes activity, including amylase and protease, significantly increased in fish fed the supplemented diets. The thyme-supplemented diets also significantly increased the biochemical parameters, including total protein, albumin, and acid phosphatase (ACP), compared to the control group. We also observed significant increases in hematological indices, including red blood cells (RBC), white blood cells (WBC), hematocrit (Hct), and hemoglobin (Hb) in common carp fed diets containing thyme oil (P < 0.05). Liver enzymes activity including alanine aminotransferase (ALT), alkaline phosphatase (ALP), and aspartate aminotransferase (AST) was also reduced (P < 0.05). Immune parameters, including total protein and total immunoglobulin (total Ig) levels, alternative complement pathway hemolytic (ACH50), lysozyme, protease, and ALP in the skin mucus, and lysozyme, total Ig, and ACH50 in the intestine were higher (P < 0.05) in TVO-supplemented fish. Catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), and glutathione peroxidase (GPx) in the liver were also elevated (P < 0.05) in TVO administered groups. Lastly, thyme-supplementation resulted in higher survival rates after the A. hydrophila challenge compared to the control (P < 0.05). In conclusion, dietary inclusion of thyme oil (1 and 2%) effectively improved fish growth, immune systems, and resistance to A. hydrophila.

Effects of dietary marjoram, Origanum majorana extract on growth performance, hematological, antioxidant, humoral and mucosal immune responses, and resistance of common carp, Cyprinus carpio against Aeromonas hydrophila.

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.

The effects of combined inclusion of Malvae sylvestris, Origanum vulgare, and Allium hirtifolium boiss for common carp (Cyprinus carpio) diet: Growth performance, antioxidant defense, and immunological parameters.

The present study aimed to investigate the effects of a mixture of three medicinal plants extracts (COP: common mallow (Malvae sylvestris), oregano (Origanum vulgare), and Persian shallot (Allium hirtifolium boiss)) on growth, hematological indices, immunological parameters, liver antioxidant, and digestive enzyme activity of common carp (Cyprinus carpio). Accordingly, 540 common carp fingerlings were randomly allotted to 18 fiberglass tanks, including six treatments each in triplicate. Fish were fed with experimental diets supplemented with 0, 0.5, 1, 2, 3, and 5% of the herbal mixture. After 60 days, the effects on growth performance, antioxidant, and immunological parameters were evaluated. According to the results, the highest final weight was observed in the fish fed 2 and 3% of the herbal mixture. Also, the feed conversion ratio (FCR) showed a significant decrease in fish treated with the herbal mix, so that the lowest FCR was observed in the treatment 1%. Hematological examinations such as red blood cells (RBC), white blood cell (WBC), and hemoglobin showed significant changes between different parameters compared to the control group. The highest protein content was observed in the fish supplemented with 2, 3, and 5% herbs. Besides, in contrast to glucose, the cortisol level showed a significant decrease compared to the control group. The digestive enzymes (Amylase, Lipase, and Protease) were significantly higher in 2 and 3% treatments than in the control group. Also, a significant difference was found in Superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) activity, and malondialdehyde (MDA) content between the experimental and control treatments. Examination of liver enzymes (and alanine aminotransaminase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP)) showed a significant difference in herbal mixture treatments with the control group, according to which the lowest amount of these enzymes was recorded in 2 and 3% herb supplemented fish. Herbal mix resulted in higher mucosal and intestinal immunity parameters, including total immunoglobulin (total Ig), lysozyme, alternative complement activities (ACH50), protease, and ALP. According to the results, supplementing the diet with a mixture of the COP can significantly improve growth parameters, stimulate the innate immune system, and improve antioxidant defenses and liver health of common carp.