Probiotics enhance resistance to Streptococcus iniae in Nile tilapia (Oreochromis niloticus) reared in biofloc systems.

Biofloc technology is a rearing technique that maintains desired water quality by manipulating carbon and nitrogen and their inherent mixture of organic matter and microbes. Beneficial microorganisms in biofloc systems produce bioactive metabolites that may deter the growth of pathogenic microbes. As little is known about the interaction of biofloc systems and the addition of probiotics, this study focused on this integration to manipulate the microbial community and its interactions within biofloc systems. The present study evaluated two probiotics (B. velezensis AP193 and BiOWiSH FeedBuilder Syn 3) for use in Nile tilapia (Oreochromis niloticus) culture in a biofloc system. Nine independent 3785 L circular tanks were stocked with 120 juveniles (71.4 ± 4.4 g). Tilapia were fed for 16 weeks and randomly assigned three diets: a commercial control diet or a commercial diet top-coated with either AP193 or BiOWiSH FeedBuilder Syn3. At 14 weeks, the fish were challenged with a low dose of Streptococcus iniae (ARS-98-60, 7.2 × 107 CFU mL-1 , via intraperitoneal injection) in a common garden experimental design. At 16 weeks, the fish were challenged with a high dose of S. iniae (6.6 × 108 CFU mL-1 ) in the same manner. At the end of each challenge trial, cumulative per cent mortality, lysozyme activity and expression of 4 genes (il-1ß, il6, il8 and tnfα) from the spleen were measured. In both challenges, the mortalities of the probiotic-fed groups were significantly lower (p < .05) than in the control diet. Although there were some strong trends, probiotic applications did not result in significant immune gene expression changes related to diet during the pre-trial period and following exposure to S. iniae. Nonetheless, overall il6 expression was lower in fish challenged with a high dose of ARS-98-60, while tnfα expression was lower in fish subjected to a lower pathogen dose. Study findings demonstrate the applicability of probiotics as a dietary supplement for tilapia reared in biofloc systems.

Effects of two host-associated probiotics Bacillus mojavensis B191 and Bacillus subtilis MRS11 on growth performance, intestinal morphology, expression of immune-related genes and disease resistance of Nile tilapia (Oreochromis niloticus) against Streptococcusiniae.

The intensification and diversification of production systems have increased the incidence of diseases, which are usually treated with antibiotics. However, its use should be restricted due to the increasing prevalence of antibiotic-resistant bacteria. Probiotics represent therefore an alternative environmentally friendly strategy for improving growth and disease resistance in aquaculture. Considering that host-derived probiotics may offer greater advantages than those from other environments in terms of safety and efficacy, two potential host-associated probiotic strains (Bacillus mojavensis B191 and Bacillus subtilis MRS11) were used in the present study, which were previously isolated from intestinal mucus of Nile tilapia (Oreochromis niloticus). This study was conducted to assess the effects of dietary administration of two Bacillus strains on growth performance, intestinal morphology, immunity, and disease resistance of Nile tilapia. A total of 375 fish were randomly divided into five groups in triplicate. Nile tilapia were fed a basal diet (control group) or a basal diet supplemented with Bacillus mojavensis B191 (BM) or Bacillus subtilis MRS11 (BS) spores at different concentrations of 1 × 106 (BM6 and BS6, respectively) and 1 × 108 (BM8 and BS8, respectively) CFU/g of feed for 60 days. Moreover, the survival rate of tilapia upon challenge with Streptococcus iniae was determined following the feeding trial. After the feeding trial, the growth performances were significantly improved in all probiotic-fed groups, with the BS8 group being the highest. Light and electron microscopy observations revealed elevated goblet cells, intestinal villus length (except BM8), microvilli length, microvilli density, and perimeter ratio increase in the intestine of all probiotic-fed groups compared with the control group. Regarding the expression analysis, HSP70 gene was only up-regulated in the BM8 group and a general trend of up-regulation of some immune-related cytokines (TGF-ß, IL-10, TNF-α and IL-1ß) was observed in all probiotic-fed groups. Likewise, the best protection against Streptococcus iniae was observed in the BS8 group, followed by BS6, BM6 and BM8 groups. Altogether, dietary probiotic supplementation with BS8 and BM6 may improve growth performance, intestinal morphology, immunity, and disease resistance in Nile tilapia.

Mooseer (Allium hirtifolium) boosts growth, general health status, and resistance of rainbow trout (Oncorhynchus mykiss) against Streptococcus iniae infection.

In large-scale aquaculture, the fast growth rate of fish is positively influenced by feed additives such as medicinal plants. This is however; infectious disease may reduce fish growth and cause devastating economic loss. The present study investigated in vitro antibacterial efficacy of Mooseer (Allium hirtifolium) extract against Streptococcus iniae and its in vivo effects on growth, biochemical parameters, innate immunity of rainbow trout (Oncorhynchus mykiss). Therefore, six experimental diets were designed to include different levels of Mooseer from zero (as control), 5, 10, 15, 20, and 25 g per kg diet respectively referred to as M1 to M5. Results from the antibacterial evaluation showed that Mooseer extract inhibits S. iniae growth with MIC and MBC values of 128 and 256 µg ml-1. Appreciable results were obtained in the groups supplemented with Mooseer. Mooseer enhanced growth performance, and modulated serum biochemical and immunological parameters (total protein, albumin, triglyceride, glucose, cortisol, cholesterol, lysozyme, Ig, ACH50, ALP, and protease activity), and liver enzymes (ALT, AST and ALP). The greatest effects were found for higher doses of Mooseer supplementation (M4 and M5). Meanwhile, results from the survival rate of fish challenged with S. iniae showed higher survival in M2 and M4 treatments. The present findings suggest the beneficial use of Mooseer in rainbow trout diet, with 20 g kg-1 inclusion as the recommended dose.

Antibacterial Activities of Prenylated Isoflavones from Maclura tricuspidata against Fish Pathogenic Streptococcus: Their Structure-Activity Relationships and Extraction Optimization.

Streptococcus zoonotic bacteria cause serious problems in aquaculture with clinical effects on humans. A structure-antibacterial activity relationships analysis of 22 isoflavones isolated from M. tricuspidata (leaves, ripe fruits, and unripe fruits) against S. iniae revealed that prenylation of the isoflavone skeleton was an important key for their antibacterial activities (minimum inhibitory concentrations: 1.95-500 µg/mL). Through principal component analysis, characteristic prenylated isoflavones such as 6,8-diprenlygenistein (4) were identified as pivotal compounds that largely determine each part's antibacterial activities. M. tiricuspidata ripe fruits (MTF), which showed the highest antibacterial activity among the parts tested, were optimized for high antibacterial activity and low cytotoxicity on fathead minnow cells using Box-Behnken design. Optimized extraction conditions were deduced to be 50%/80 °C/7.5 h for ethanol concentration/extraction temperature/time, and OE-MTF showed contents of 6,8-diprenlygenistein (4), 2.09% with a MIC of 40 µg/mL. These results suggest that OE-MTF and its active isoflavones have promising potential as eco-friendly antibacterial agents against streptococcosis in aquaculture.

Supplementation of zebrafish (Danio rerio) diet using a short antimicrobial peptide: Evaluation of growth performance, immunomodulatory function, antioxidant activity, and disease resistance.

Short-chain bioactive peptides are new and promising antimicrobial, immune moderating, and antioxidant agents. Therefore, the present study was conducted to evaluate in vitro antibacterial activity of CM11, a short antimicrobial peptide (AMP), against Streptococcus iniae and Yersinia ruckeri as fish pathogenic bacteria using standard disk diffusion and microdilution assays. In addition, in vivo effects of CM11 on fish growth, immunity, antioxidant activity, and disease resistance were evaluated using zebrafish (Danio rerio) as an animal model. For in vivo study, based on in vitro susceptibility results, four diets were designed to include zero (as control), 10, 20, and 50 µg of CM11 per g diet referred to as control, P1, P2, and P3 treatments, respectively. After eight weeks of dietary trial, fish were challenged with Streptococcus iniae, and the survival rate was calculated for a period of two weeks. Results showed that CM11 effectively inhibited the growth of S. iniae and Y. ruckeri on agar plates at a concentration of eight µg/ml. Minimum inhibitory and minimum bactericidal concentrations of CM11 were measured at 8 and 32 µg/ml for S. iniae and 16 and 64 µg/ml Y. ruckeri, respectively. In vivo results showed no noticeable effects on fish growth parameters, however, feed conversion ratio (FCR) was found lower in P3 and P2 compared to control (P < 0.05). Immunological and antioxidant responses were found strongly affected by CM11 in all treatment groups in which the highest values were found in the P3 treated group. Key immune and antioxidant genes were up-regulated particularly in fish receiving the highest level of CM11 (P3). Fish receiving the CM11 peptide showed better survival when challenged with S. iniae. These findings suggest the potential of CM11 for use in aquaculture as an antibacterial and immunostimulant agent.

Dietary supplementation with dandelion (Taraxacum officinale) flower extract provides immunostimulation and resistance against Streptococcus iniae infection in rainbow trout (Oncorhynchusmykiss).

Dandelion flower is a well-known phytomedicine due to its therapeutic effects on dyspepsia, bile duct disorders, spleen and liver complaints, and inflammatory diseases. This study aimed to assess the effect of dietary dandelion flower extract (DFE) on skin mucosal and humoral immunity, immune-related gene expression, and disease resistance in rainbow trout. The fingerlings (N = 300, 20.1 ± 0.9 g) were randomly assigned to 15 tanks and fed with different levels of DEF including 0 (control), 1, 2, 3, and 4 g/kg for 56 days. At the end of the trial, the total leukocyte and lymphocyte counts were significantly enhanced in DFE-added groups compared to the control group. Complement components, immunoglobulin M, total protein, and lysozyme showed significantly higher values in the fish treated with dietary DFE at 3 g/kg than others (P < 0.05). The enzymatic activities of the skin mucus samples in the fish fed dietary levels of DFE were significantly higher than the control fish. Skin mucus protein was also improved significantly in 2 and 3 g/kg DFE compared to others. The transcription levels of interleukin-1ß and interleukin-6 genes were up-regulated in the fish fed with 3 and 4 g/kg DFE. Interleukin-8 and lysozyme gene expression levels were elevated in 3 g/kg DFE group compared to the control group. Fish mortality after challenging with Streptococcus iniae was significantly reduced from 43.33% in the control group to 73.33% in 3 g/kg DFE group. This study confirmed the beneficial effects of DFE as an immunostimulant in rainbow trout diet and the recommended dose of DFE is 2.49-2.74 g/kg based on the polynomial regression models.

Dietary effects of a low-molecular weight fraction (<10 kDa) from shrimp waste hydrolysate on growth performance and immunity of rainbow trout (Oncorhynchus mykiss): Employing nanodelivery systems.

Aquaculture by-products have been of great interest for producing protein hydrolysates with multiple biological activities. The present experiment was carried out to evaluate dietary effects of a low-molecular fraction (<10 kDa) from shrimp waste hydrolysate in forms of unprotected and nanocapsulated on growth and immunity of rainbow trout. Therefore, six diets were designed including a control diet (no supplementation), D1 (1 g kg-1 of unprotected fraction), D2 (1 g kg-1 chitosan nanocapsules), D3 (1 g kg-1 liposome nanocapsules), D4 (1 g kg-1 of fraction-loaded chitosan nanocapsules), D5 (1 g kg-1 of fraction-loaded liposome nanocapsules). Fish (0.91 ± 0.15 g) were fed with experimental diets until apparent satiation for six weeks followed by a 5-day experimental challenge with Streptococcus iniae. Results revealed that growth is strongly affected in fish receiving the fraction with D4 treatment showing the highest weight gain, SGR, final weight and the lowest FCR (p < 0.05). Nanocapsules without fraction did not show remarkable effects when compared to control group. In terms of serum and mucus immune parameters of lysozyme, complement activity, myeloperoxidase activity, and total protease, fish from D4 group showed the highest measured values followed by D5 (p < 0.05). Key immune related genes of IL-6 and TNF-α were noticeably up-regulated in fish from D1, D4, and D5 groups, which were consistent with survival rate after 5 days challenge with Streptococcus iniae. All together, the present findings highlighted the application of chitosan and liposome nanocarriers in aquaculture and potential of low-molecular weight fraction (<10 kDa) from shrimp wastes hydrolysate to improve growth performance and immune status of rainbow trout.

Herbal hydrogel-based encapsulated Enterococcus faecium ABRIINW.N7 improves the resistance of red hybrid tilapia against Streptococcus iniae.

AIMS: The streptococcal disease has been associated with serious mortality and significant global economic loss in the tilapia farming industry. The overall goal of this work was to test herbal hydrogels based on encapsulated Enterococcus faecium ABRIINW.N7 for potential probiotic anti-microbial activity against Streptococcus iniae in red hybrid tilapia. METHODS AND RESULTS: Abnormal behaviour, clinical signs, postinjection survival and histopathology (kidney, liver, eye and brain) were measured. Cumulative mortality of CON+ , free cells, ALG and treatments (F1-F7) was 30, 24, 22, 19, 17, 17, 16, 14, 14 and 12 out of 30 fish and the survival rates for E. faecium ABRIINW.N7 microencapsulated in an alginate-BS blend with 0·5, 1, 1·5, 2, 2·5 and 3% fenugreek were 43, 43, 47, 53, 53 and 60%, respectively. After the incorporation of fenugreek with the alginate-BS blend, there was an 8-21% increase in probiotic cell viability. Furthermore, the survival rate for the alginate-BS blend with 2·5 and 3% fenugreek (F6 and F7) was significantly (P ≤ 0·05) higher than other blends. The highest encapsulation efficiency, viability in gastrointestinal conditions and during storage time and excellent antipathogenicity against S. iniae were observed in alginate-BS +3% fenugreek formulation (F7). CONCLUSIONS: It is recommended that probiotic strains like E. faecium ABRIINW.N7 in combination with local herbal gums, such as BS and fenugreek plus alginate, can be used as a suitable scaffold and an ideal matrix for the encapsulation of probiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study proposes models connecting process parameters, matrix structure and functionality.

Cutaneous mucosal immune-parameters and intestinal immune-relevant genes expression in streptococcal-infected rainbow trout (Oncorhynchus mykiss): A comparative study with the administration of florfenicol and olive leaf extract.

This study evaluated changes in cutaneous mucosal immunity (total protein (TP) and immunoglobulin (TIg), lysozyme, protease, esterase, and alkaline phosphatase (ALP)) and some immune-related genes expression (tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-8, hepcidin-like antimicrobial peptides (HAMP), and immunoglobulin M (IgM)) in the intestine of rainbow trout (Oncorhynchus mykiss) orally-administrated florfenicol (FFC) and/or olive leaf extract (OLE), experimentally infected with Streptococcus iniae. The juvenile fish (55 ± 7.6 g) were divided into different groups according to the use of added OLE (80 g kg-1 food), the presence/absence of FFC (15 mg kg-1 body weight for 10 consecutive days), and the streptococcal infectivity (2.87 × 107 CFU mL-1 as 30% of LD50-96h). The extract's chemical composition was analyzed using the high-performance liquid chromatography (HPLC) system. The skin mucus and intestine of fish were sampled after a 10-day therapeutic period for all groups, and their noted indices were measured. Our results signified that the oleuropein, quercetin, and trans-ferulic acid were the most obvious active components of OLE which were found by HPLC analysis. The combined use of OLE and FFC could lowered some skin mucus immunological indices (e.g., TP, TIg, and ALP), and the gene expression of inflammatory cytokines (e.g., TNF-α and IL-1ß) of rainbow trout. Moreover, lysozyme and protease activities respectively were invigorated by the FFC and OLE treatment. Also, the use of OLE as a potential medicine induced the gene expression of HAMP. As the prevention approach, it would be recommended to find the best dose of OLE alone or in combination with the drug through therapeutics period before the farm involved in the streptococcal infection.

Withania somnifera dietary supplementation improves lipid profile, intestinal histomorphology in healthy Nile tilapia (Oreochromis niloticus), and modulates cytokines response to Streptococcus infection.

Despite Withania somnifera (WS), stimulating effects have been investigated on many animal species, its role on lipid profile and intestinal histomorphology in healthy animals, and its modulating role on pro-inflammatory cytokines following infection in fish are yet scarce. In this context, lipid profile, liver, and intestinal histomorphology were measured in Nile tilapia fed with a basal diet or diets containing 2.5 and 5% of supplementary WS for 60 days. Besides, cytokines response was measured at 1, 3,7, and 14 days following Streptococcus iniae (S. iniae) infection after the feeding trial. All lipid profile parameters were nominally lowered, excluding high-density lipoprotein (HDL) that exhibited a significant increase in WS 5% group compared to other groups. Improved gut health integrity was observed, especially in WS 5% group in terms of increased goblet cell numbers, villous height, the width of lamina propria in all parts of the intestine, and a decrease in the diameter of the intestinal lumen of the distal intestine only. A significant down-regulation in the mRNA transcript level of cytokine genes (interleukin 1ß/IL-1ß, tumor necrosis factor α/TNFα, and interleukin 6/IL-6) was demonstrated in the kidney and spleen of WS-supplemented groups following S. iniae infection compared with the control infected (positive control/PC) group. Our findings give new insights for the potential roles of WS dietary inclusion not only on lipid profile and intestinal health integrity improvement in healthy fish under normal rearing but also as a prophylactic against the infection. Thus, WS can be incorporated as a promising nutraceutical in aquaculture.

Therapeutic and Histopathological Effect of Aloe vera and Salvia officinalis Hydroethanolic Extracts against Streptococcus iniae in Rainbow Trout.

Some of the medicinal plants have antibacterial contents and appear to be proper alternatives for antibiotics in the treatment of streptococcal disease, which causes plenty of mortalities in fish farms annually. Therefore, this study investigated the therapeutic effect of Aloe vera and Salvia officinalis hydroethanolic extracts against Streptococcus iniae in rainbow trout. Plant extracts components were analyzed by Gas chromatography-mass spectrometry method and tested in vitro against S. iniae by disk diffusion assay. In in vivo, 480 rainbow trout (10&plusmn;0.1 g) were distributed in 9 groups (with 3 replication), and all groups (except for the first group as the negative control that was injected with 100 &micro;l of physiologic serum) were injected by 100 &micro;l of LD50 (3.66&times;108.5CFU/ml) of S. iniae suspension, intraperitoneally. The fish of groups were treated by A. vera and S. officinalis extracts in doses 0 (positive control group was fed by commercial diet without plant extract), 0.5, 1, and 1.5% (supplemented diet) and 80 mg/kg body weight erythromycin for the next 10 days. At the end of the study period, tissue samples of the gills and livers of all groups were taken for the histopathological lesion assay. The results showed that A. vera and S. officinalis had antibacterial components as Cineol, and S. iniae was sensitive to both A. vera (MBC=4.067 mg/ml) and S. officinalis (MBC=5.185 mg/ml) extracts. At the end of the treatment period, there were no significant differences among erythromycin, A. vera (1.5%), and A. vera (1%) in terms of the mortality of the infected fish (P˂0.05). Moreover, A. vera (1.5%) showed a significantly lower mortality rate, compared to the positive control (P˂0.05). A. vera (1.5%) was the best group to moderate all histopathological lesions, compared to other groups. Accordingly, A. vera (1.5 %) is useful to treat streptococcosis (caused by S. iniae) and alter gill and liver histopathological lesions in rainbow trout.

Antibacterial and antioxidant activity of clove oil against Streptococcus iniae infection in Nile tilapia (Oreochromis niloticus) and its effect on hepatic hepcidin expression.

This study was designed to evaluate the modulating effect dietary clove essential oil (CL) has on the antioxidant and immunological status of Nile tilapia following Streptococcus iniae (Si) infection. Fish were placed on either control or (1.5 and 3%) CL-supplemented diets for 4 weeks. After sampling, the remaining fish in the control group were divided into 2 groups: an unchallenged (negative control) and an Si-challenged positive control. On the other hand, the remaining fish in CL-supplemented groups were challenged with Si, and mortality was checked for two weeks before the final sampling. Serum immunological parameters, tissue antioxidants, and oxidative stress markers were determined. Moreover, hepatic hepcidin expression was also measured in different groups. The obtained results showed improvements in blood phagocytic, bactericidal, lysozyme, and respiratory burst activities in CL-supplemented fish before and after the Si challenge. Si-challenge caused a remarkable increase in tissue malondialdehyde (MDA) levels that was inhibited by CL supplementation. The activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD) in tissues were significantly elevated in a dose-dependent manner in CL-supplemented groups in both pre- and post-challenge experiments; renal SOD did not show any differences. Hepatic nitric oxide (NO) level was significantly decreased in CL-supplemented fish in a dose-dependent manner. In the post-challenge experiment, nitrosative stress was apparent in the liver and kidney; however, CL supplementation was sufficient to reverse it. Interestingly, a remarkable induction of the hepatic hepcidin expression was observed in all CL-supplemented groups in the pre-challenge experiment and Si-challenged fish, underscoring the role of CL as an antibacterial through inducing hepatic hepcidin expression to combat S. iniae infection. CL-supplementation was associated with lower mortality rates after Si-challenge, which was more pronounced in CL-3% supplemented fish. In conclusion, our results demonstrate that CL has a potent antioxidant role via increasing antioxidant enzymes' activities and antagonizing lipid peroxidation. Moreover, CL has an immune-stimulant effect by inducing the hepatic hepcidin expression and immunological markers in response to S. iniae infection.

Effects of dietary non-viable Bacillus sp. SJ-10, Lactobacillus plantarum, and their combination on growth, humoral and cellular immunity, and streptococcosis resistance in olive flounder (Paralichthys olivaceus).

Heat-killed (HK) Bacillus sp. SJ-10 (B), HK Lactobacillus plantarum (P), and their combination were dietary supplemented to olive flounder (Paralichthys olivaceus) to quantify the effects on growth, innate immunity, and disease resistance. Four test diets were supplied: a control feed free of HK probiotics, 1 × 108 CFUs g-1 single treatments of each of HK B (HKB) and HK P (HKP), and an equal proportion of (0.5 HKB + 0.5 HKP) × 108 CFUs g-1 (HKB0.5 HKP0.5). At 8 weeks of completion feeding trail, HKB0.5 HKP0.5 significantly (P < .05) improved growth, feed utilization, and nonspecific immune parameters (respiratory burst and superoxide dismutase) compared to the control group. Similarly, serum lysozyme and myeloperoxidase activities were higher in both HKB and HKB0.5HKP0.5 groups. The levels of pro-inflammatory cytokine IL-6 in the liver and IL-1ß in the liver, kidney, and spleen were also improved in the treatments, but microvilli length was only increased in HKB0.5HKP0.5. After Streptococcus iniae 1 × 108 CFUs mL-1 challenged; HKB and HKB0.5HKP0.5 had a higher survival than control and HKP. Overall, dietary administration of synergy HK probiotics elevated growth, cellular and humoral immunity, and streptococcosis resistance in olive flounder.

High efficacy and economical procedure of oral vaccination against Lactococcus garvieae/Streptococcus iniae in rainbow trout (Oncorhynchus mykiss).

The present study was aimed to examine the efficacy of chitosan-alginate coated vaccines against pathogenicity of Lactococcus garvieae and Streptococcus iniae in rainbow trout. Fish were divided into four groups including: Group A: fish immunized by chitosan-alginate coated vaccine, Group B: fish immunized by non-coated vaccine, Group C: fish feed by chitosan-alginate coated pellets without vaccine and Group D: fish feed by basic diet (non-coated and without vaccine). In groups A and B, the vaccination was carried out for 14 days and after that supplemented with fundamental diet (control diet). Comparable to groups A and B, fish of group C were also fed 14 days with test diets and after that fed control food. On day 0, 20, 40 and 60 of the experiment, serum samples were given. Fish have been challenged with live L. garvieae and S. iniae after 60 days. The levels of bactericidal activity and complement activity among innate immunity components extended on day 20 of the research and after that decreased in group A and B (P < 0.05) all through the examination. The relative expression of IL-6 and IgM in groups A and B extended on examination day 20. The expression of these genes illustrated no advancements in different groups in during the examination (P > 0.05). In group A, the serum antibody titer against L. garvieae and S. iniae broadly raised on day 40 and 60 of examination, whereas in group B, the immune response titer against S. iniae and L. garvieae illustrated a significant elevation on day 60 of the trial (P < 0.05). After challenge with live bacteria, survival rate of 83 ± 9.1%(challenged with S. iniae) and 72.18 ± 9.8% (challenged with L. garvieae) were gotten independently in group A, which were higher than survival of other exploratory groups (P < 0.05). In conclusion, the results of the present examination appear that the orally vaccination of rainbow trout with chitosan-alginate covered vaccine stimulates immunity system and also efficiently protects rainbow trout against Lactococcus garvieae and Streptococcus iniae.

Antibacterial activity of Oliveria decumbens against Streptococcus iniae in Nile tilapia (Oreochromis niloticus) and its effects on serum and mucosal immunity and antioxidant status.

The aims of this study were to investigate the antibacterial, immunostimulatory and antioxidant properties of different derivatives of Oliveria decumbens, in vitro and in vivo. The GC-MS spectrometry analysis showed γ-terpinene as the most frequent compound in essential oil, whereas carvacrol and thymol were the most common ones in aromatic water. Plant essential oil and hydroethanolic extract showed a positive in vitro bactericidal activity against Streptococcus iniae as evaluated by disc diffusion, minimum inhibitory concentration and minimum bactericidal concentration methods. Also, in vivo resistance against S. iniae and immune and antioxidant responses of Nile tilapia (Oreochromis niloticus) were assayed after 60 days treatment with O. decumbens derivatives. Plant hydroethanolic extract and essential oil and their 1:1 combination were added to diet at 0 (negative control), 0.01, 0.1 and 1% (w:w). The plant aromatic water at doses of 0.0312, 0.0625 and 0.1250% were also used as bath treatment. The results showed that aromatic water at lowest dose was more effective than other treatments in increment of fish resistance against S. iniae (7.14% mortality in comparison with 50% mortalities in control fish) and modulation of post-challenge respiratory burst activity. The bactericidal activity and biochemical contents of skin mucus did not change significantly among treatments. The levels of superoxide dismutase and catalase antioxidant enzymes activities in spleen tissue were significantly higher in treatments received extract, essential oils and their combination in comparison to other groups, while treatments did not affect peroxidase level. In conclusion, administration of different derivatives of Oliveria decumbens showed remarkable antibacterial activity against streptococcosis and enhanced antioxidant status and post-challenge immunity in Nile tilapia.

Efficacy of dietary Nano-selenium on growth, immune response, antioxidant, transcriptomic profile and resistance of Nile tilapia, Oreochromis niloticus against Streptococcus iniae infection.

As recently applicable, there are few studies on the impact of using nano-selenium (nano-Se) on varied fish species. Where nothing reachable focused on its impact on tilapias so, the present analysis evaluated the efficacy of using nano-Se in tilapias on immune response, antioxidant defense compared by conventional Se form. 480 O. niloticus fingerlings were haphazardly grouped firstly into three groups with four replicates of each. The control one (CT) was fed on a basal diet. The second and third one supplemented with 0.7 mg/kg-1 Se and nano-Se respectively for ten weeks. At the start day of the ninth week, two replicates from each group were injected by Streptococcus iniae where, the remaining replicates stand without challenge. Enhancement of growth performance measurements were noted in nano-Se compared to Se or CT groups. Existed anemia in S. iniae tilapias became alleviated by using nano-Se that also, improves the alteration of leucogram induced by challenge. Elevation of aminotransferases, alkaline phosphatase, lactate dehydrogenase (ALT, AST, ALP and LDH) and creatinine in Se and CT challenged replicates that seemed nearly normal by using nano-Se. Usage of nano-Se showed more powerful antioxidant activities than Se. There were an expansion of immunoglobulin M, lysozymes, glutathione peroxidase, nitric oxide, superoxide dismutase and catalase (IgM, LYZ, GPx, NO, SOD, CAT) and their related gene expression in nano-Se with contrast in Se or CT challenged groups. Nile tilapias challenged by S. iniae disclosed substantial expansion in the percentage of mortality in CT challenged fish (93.33%), followed by the group supplemented with Se (73.33%), whereas the lowermost one at fish supplemented by nano-Se (26.66%). The mortalities have been stopped from the 5th, 12th and 14th days in, nano-Se, Se and CT respectively. It can be concluded that using of Se 0.7 mg/kg-1induce immunosuppressive, antioxidant, liver and kidneys negative impact on tilapias where the same dose from nano-Se was more potent immunomodulating and antioxidant. Also it is attend in counteracting the serious impact induced by S. iniae challenge.

Dietary sodium butyrate (Butirex® C4) supplementation modulates intestinal transcriptomic responses and augments disease resistance of rainbow trout (Oncorhynchus mykiss).

Intestine in fish is a complex multifunctional organ, not only plays roles in digestion and absorption of nutrient, but also has critical role in immunity. The present study evaluated the effects of different levels of dietary sodium butyrate [Butirex® C4 (Butirex)] on intestinal immune-,antioxidant-and tight junction-related gene expression injuvenile rainbow trout(Oncorhynchusmykiss). 240 healthy rainbow trout were dispensed in 12 fiberglass tanks appointed to four treatments [0 (control), 1.5 (B1.5), 2.5 (B2.5) and 5 (B5)g Butirex per kg diet]. After a 45-day feeding trial, the fish fed with the Butirex-supplemented diets showed higher intestinal lysozyme (LYZ), complement(ACH50) and bactericidal activities; the elevations in ACH50 and bactericidal activities depended on Butirex levels (P < 0.05). The Butirex-supplemented groups, particularly the B2.5 group, had significantly higher LYZ gene expression compared to the control group (P < 0.05). Butirex at 2.5 and 5 g/kg levels led to significantly higher IL-1ß gene expression. B2.5 and B5 had significantly lower and higher TNF-α gene expression compared to the control group (P < 0.05). The B2.5 group had significantly higher TGF-B, and significantly lower IL-8 compared to the control group (P < 0.05). The B1.5 and B2.5 group had significantly higher IL-10 gene expression compared to the control group (P < 0.05). The B2.5 and B5 groups had significantly higher SOD gene expression compared to the other groups; the highest expression was related to the B2.5 group (P < 0.05). Dietary Butirex supplementation significantly up-regulated CAT and GPx genes expression compared to the control group; the highest expression as related to the B2.5 and B5 groups (P < 0.05). The B2.5 group had significantly lower CLD12 gene expression compared to the control group (P < 0.05). The B2.5 and B5 groups had significantly higher CLD3, OCLD and ZO-1 gene expression compared to the control. The highest CLD3, ZO-1 gene expressions was related to the B2.5, and B5 groups respectively (P < 0.05). After challenge with Streptococcus iniae, B2.5 and B5 had significantly higher survival compared to the control group (55.6 ±â€¯7.70 and 68.9 ±â€¯10.2 vs. 33.3 ±â€¯6.67). In conclusion, Butirex is efficient immune stimulant and health booster in rainbow trout, which augments the fish resistance to disease. Modulation of immune components, cytokines, antioxidant system and intestinal integrity might involve in improving disease resistance in Butirex-treated fish. Although most of the examined genes were modulated by 2.5 g/kg Butirex under normal conditions, 5 g/kg level is recommended under pathogenic state to mitigate mortality.

Dietary vitamin E deficiency inhibits fat metabolism, antioxidant capacity, and immune regulation of inflammatory response in genetically improved farmed tilapia (GIFT, Oreochromis niloticus) fingerlings following Streptococcus iniae infection.

Vitamin E plays an important role in maintaining normal metabolism and physiological functions in animals. The health of fish fingerlings directly affects the rate of disease incidence in adult fish, and healthy fingerlings ultimately result in better breeding outcomes for cultured fish. To date, no previous studies have focused on the effects vitamin E deficiency on tilapia at the fingerling stage. In this study, we investigated the effects of dietary vitamin E on the growth, fat metabolism, antioxidant capacity, and inflammatory response of genetically improved farmed tilapia (GIFT, Oreochromis niloticus) fingerlings. Vitamin E at different concentrations (0, 20, 40, 80, 160, and 320 mg/kg) was added to the diet and GIFT were fed for 55 days. Then, the GIFT were intraperitoneally injected with Streptococcus iniae and tested for infection. Vitamin E deficiency decreased growth and increased the food conversion ratio of GIFT fingerlings. Vitamin E deficiency also reduced the white blood cell count, increased hematocrit and hemoglobin contents in the blood, increased serum aspartate aminotransferase and alanine aminotransferase activities, and increased liver stress (P < 0.05). Vitamin E deficiency inhibited fat metabolism, down-regulated the expression of genes encoding lipoprotein lipase and heart-type and liver-type fatty acid-binding proteins, and increased serum total protein and fat deposition. Vitamin E deficiency significantly decreased superoxide dismutase, glutathione peroxidase, and catalase activities, increased malondialdehyde content, and caused oxidative damage. Vitamin E deficiency also up-regulated the expression of genes encoding interleukin 1ß and tumor necrosis factor α in the head kidney, and stimulated a pro-inflammatory response. Overall, vitamin E deficiency inhibited growth, impaired fat metabolism, and disrupted the inflammatory response of GIFT fingerlings, whereas vitamin E supplementation in the diet reversed these negative effects. The diets with high concentrations of vitamin E (160-320 mg/kg) led to vitamin E accumulation in the fish tissues and rapid activation of the inflammatory response and antioxidant capacity in GIFT fingerlings exposed to S. iniae.

Effects of dietary hydrolyzed yeast (Rhodotorula mucilaginosa) on growth performance, immune response, antioxidant capacity and histomorphology of juvenile Nile tilapia (Oreochromis niloticus).

An 8 weeks feeding experiment was conducted to evaluate the effects of dietary supplementation with hydrolyzed yeast (HY) (Rhodotorula mucilaginosa) on growth performance, hematological parameters, immune response and antioxidant ability of juvenile Nile tilapia. Five isonitrogenous and isolipidic diets (32% protein and 4% lipid) with different levels (0%, 0.125%, 0.25%, 0.5%, 1%) of HY were formulated. Each diet was randomly assigned to quadruplicate groups of fish (initial body weight 19.1 ±â€¯0.01 g). Results indicated that significantly higher specific growth rate (SGR) and lower feed conversion rate (FCR) were obtained in fish fed 1% HY diet than that of fish fed 0% HY diet (P < 0.05). Fish fed 0.25% HY diet showed the lowest value of hepatopancreas somatic indices (HSI) and significantly lower than that of fish fed 0% HY diet (P < 0.05). Meanwhile, protein and ash in the whole-body content of fish fed 1% HY diet was significantly higher than that of fish fed 0%-0.5% HY diets. Serum immunological parameters showed that the lysozyme (LZM) activity and Complement C3 content were significantly increased by dietary supplementation of 0.5%-1% HY (P < 0.05). However, dietary supplementation with 0.125%-1% HY significantly decreased the activity of myeloperoxidase (MPO) (P < 0.05). Antioxidant status in serum and liver was significantly enhanced by dietary supplementation of 0.25%-1% HY through the remarkably improved superoxide dismutase (SOD) activity both in serum and liver, the raised total antioxidative capacity (T-AOC) of serum as well as the notably reduced malondialdehyde (MDA) content in the liver (P < 0.05). However, T-AOC in the liver was not significantly influenced among all diet treatments (P > 0.05). Villi height and intraepithelial lymphocytes (IEFs) of mid-intestine were significantly higher in fish fed 0.5%-1% HY diets (P < 0.05). The challenge test demonstrated the enhanced protection against Streptococcus iniae strain by the obtained higher cumulative survival rate. In conclusion, dietary supplementation of 1% HY could maintain the better growth performance, nutrient composition as well as immune response and antioxidant capacity for juvenile Nile tilapia.

Heat-killed Bacillus sp. SJ-10 probiotic acts as a growth and humoral innate immunity response enhancer in olive flounder (Paralichthys olivaceus).

Investigations were carried out to evaluate and quantify the effects of dietary supplementation with heat-killed (HK) Bacillus sp. SJ-10 (BSJ-10) probiotic (1 × 108 CFU g-1) on the growth and immunity of olive flounder (Paralichthys olivaceus). Flounder (averagely 9.64 g) were divided into two groups, and fed control and HK BSJ-10 (HKBSJ-10)-inoculated diets for 8 weeks. Investigations were carried out on growth and feed utilizations, innate immunity, serum biochemical parameters, microvilli length, and pro- and anti-inflammatory cytokine gene (tumor necrosis factor [TNF]-α, interleukin [IL]-1ß, IL-6, and IL-10) transcriptions. Compared to control, HKBSJ-10 diet significantly (P < 0.05) enhanced weight gain and protein efficiency ratio, 1.17 and 1.11 folds respectively. Humoral innate immune parameters, lysozyme and superoxide dismutase in treatment group were also elevated by 1.34 and 1.16 folds. Similarly, an increased (P < 0.05) relative expressions of TNF-α, IL-1ß, IL-6 were recorded in liver (2.71, 3.38, and 4.12 folds respectively), and gill (2.08, 1.98, and 1.81 folds respectively) than that of controls. Moreover, after challenge with Streptococcus iniae (1 × 108 CFU mL-1), the HKBSJ-10-fed group exhibited significantly higher protection (P < 0.05) against streptococcosis compared to controls, validating the observed changes in immune parameters and induction on the cytokine-encoding genes. Therefore, HKBSJ-10 increases growth, modulates innate immune parameters, and protects olive flounders against streptococcosis.