Effects of supplementation with different zinc-based products on the growth and health of Nile tilapia.

Zinc is one of the essential microelements for the metabolism of animals. Zinc nanoparticles may have higher bioavailability due to their low specific surface area, facilitating absorption by fish. The present study aimed to evaluate the effects of supplementation with different zinc-based products on the growth and health of Nile tilapia Oreochromis niloticus. Zinc, in different sizes (nanoparticles or bulk) and forms (inorganic or organic), were used as a supplement in the tilapia diet at a dose of 15 mg kg feed-1 for 60 days. At the end of the feeding trial, production performance, hemato-immunological parameters, activity of antioxidant system enzymes, exposure to Streptococcus agalactiae and zinc concentration in the muscle were examined. After the bacterial challenge, the mean corpuscular hemoglobin concentration (MCHC) significantly increased in the fish treated with organic zinc, inorganic nano zinc, and organic nano zinc, while in the control group (inorganic zinc), MCHC remained unchanged. Regarding defense cells, dietary inorganic nano zinc increased the number of basophils (1.50 ± 1.10) compared to organic zinc (0.80 ± 0.90). Lymphocyte count increased after the challenge only in the organic zinc treatments (bulk and nanoparticles). Neutrophils decreased in the control (inorganic zinc) (2.20 ± 1.70) and inorganic nano zinc (2.60 ± 2.70) treatments after the challenge. When compared before and after the bacterial challenge, the plasma antimicrobial titer significantly increased after the bacterial challenge in all treatments. No significant differences were observed for total proteins, enzymes (SOD and CAT), cumulative survival and zinc deposition on fillet. In conclusion, organic zinc in nanoparticles or bulk size increased Nile tilapia innate defense during bacterial infection. However, the other parameters evaluated were not affected by zinc particle size or form (organic or inorganic), indicating that further evaluations should be conducted with organic zinc in nanoparticles or bulk size in the tilapia diet.

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.

Immune response and protective efficacy of two new adjuvants, Montanide™ ISA 763B VG and Montanide™ GEL02, administered with a Streptococcus agalactiae ghost vaccine in Nile tilapia (Oreochromis niloticus).

Streptococcus agalactiae is one of the most important pathogens infecting tilapia worldwide and causes meningoencephalitis, septicemia and high mortalities with considerable losses. Various types of vaccines have been developed against S. agalactiae infection, such as inactivated vaccines, live attenuated vaccines and subunit vaccines. Bacterial ghosts (BGs) are nonliving, empty cell envelopes and have been reported as novel vaccine candidates. Therefore, the main aims of this study were to develop an S. agalactiae ghost vaccine (SAGV) and to evaluate the immune response and protective effect of SAGV against S. agalactiae with two novel adjuvants, Montanide™ ISA 763B VG and Montanide™ GEL02. Nile tilapia, mean weight 50 g, were divided into four groups as follows; 1) fish injected with PBS as control, 2) fish injected with the SAGV alone; 3) fish injected with the SAGV+Montanide™ ISA 763B VG; and 4) fish injected with SAGV+Montanide™ GEL02. Following vaccination, innate immunity parameters including serum lysozyme, myeloperoxidase, catalase, and bactericidal activity were all significantly enhanced. Moreover, specific serum IgM antibodies were induced and reached their highest level 2-8 weeks post vaccination. Importantly, the relative percent survival of tilapia vaccinated against the SAGV formulated with both adjuvants was 80-93%. Furthermore, the transcription of immune-related genes (IgM, TCRß, IL-1ß, IL-8 and TNFα) were up-regulated in tilapia after vaccination, indicating that both cellular and humoral immune responses were induced by these adjuvanted vaccines. In summary, Montanide™ ISA 763B VG and Montanide™ GEL02 can enhance immunoprotection induced by the SAGV vaccine against streptococcosis, demonstrating that both have value as potential adjuvants of fish vaccines.

Impacts of pineapple peel powder on growth performance, innate immunity, disease resistance, and relative immune gene expression of Nile tilapia, Oreochromis niloticus.

An 8-week growth trial was conducted to examine the efficacy of pineapple peel powder (PAPP) on growth rate and immunity of Nile tilapia, O. niloticus. Three hundred Nile tilapia (20.91 ± 0.11 g) were fed five diets containing different levels of PAPP at 0, 10, 20, 30 and 40 g kg-1 PAPP, respectively. After four and eight weeks of the feeding trial, growth rates, and immune responses were tested. A challenge test using Streptococcus agalactiae and relative immune gene expression were performed after eight weeks of PAPP feeding. It was found that skin mucus and serum lysozyme, skin mucus and serum peroxidase, alternative complement, phagocytosis, and respiratory burst activities were significantly increased with the addition of PAPP. The maximum (P ≤ 0.05) innate immune values were noted in fish fed 10 g kg-1 PAPP. Similarly, the up-regulation of IL1, IL8, and LBP gene expressions were also detected in fish fed PAPP diets, with the maximum value was found in 10 g kg-1 PAPP fed fish. The relative percentage of survival (RPS) of Oreochromis niloticus after the challenge test were (56.00%, 72.00%, 60.00%, and 44.00%) for the 5, 10, 20 and 40 g kg-1 PAPP diets, respectively. Fish fed the 10 g kg-1 PAPP supplemented diet achieved the highest (P < 0.05) survival rate against S. agalactiae. Growth and feed efficiency were outstandingly (P < 0.05) enhanced in the PAPP groups. In conclusion, PAPP can be potentially used as a feed additive in Nile tilapia culture under Biofloc system.

Dietary cinnamaldehyde nanoemulsion boosts growth and transcriptomes of antioxidant and immune related genes to fight Streptococcus agalactiae infection in Nile tilapia (Oreochromis niloticus).

The present study was conducted to investigate the effects of dietary cinnamaldehyde nanoemulsion (CNE) on growth, digestive activities, antioxidant and immune responses and resistance against Streptococcus agalactiae (S. agalactiae) in Nile tilapia. Four experimental diets were formulated containing CNE at levels of 0, 100, 200 and 300 mg/kg diet for 12 weeks. At the end of the experiment, all fish were challenged by S. agalactiae. The results showed that the final body weight was increased in fish groups fed 200 and 300 mg CNE/kg diet by 18.4 and 17.2% with respect to the control group. Moreover, feed conversion ratio and digestive enzymes' activities were improved in groups fed 200 and 300 then 100 mg of dietary CNE/kg diet. Groups fed CNE exhibited a significant increase in serum immune-related parameters when compared with control group. Additionally, the hypocholesterolemic effects was achieved after CNE feeding unlike the control group in a dose dependent manner. With increasing dietary CNE levels, genes expression of cytokines and antioxidant enzymes were upregulated. Less severe adverse clinical symptoms and respectable cumulative mortalities associated with S. agalactiae infection were observed in fish fed CNE. To our knowledge, this study was the first offering a protective effect of CNE against S. agalactiae infection in Nile tilapia with a maximum down-regulation of cylE and hylB virulence genes expression noticed in group fed 300 mg of CNE/kg diet (up to 0.10 and 0.19- fold, respectively). Therefore, the present study recommended that an incorporation of CNE at level of 300 mg/kg diet for Nile tilapia could promote their growth, enhance their immunity and antioxidant status and provide protection against virulent S. agalactiae.

Dietary ß-glucan (MacroGard®) improves innate immune responses and disease resistance in Nile tilapia regardless of the administration period.

The effects of dietary ß-glucan on innate immune responses have been shown in a number of different vertebrate species. However, there is conflicting information about the period of administration (shorter vs. longer), and it is also unclear to what extent ß-glucan's effects can be observed post-treatment in fish. Thus, we fed Nile tilapia for 0 (control group; 45 days of control diet), 15 (30 days of control followed by 15 days of ß-glucan), 30 (15 days of control followed by 30 days of ß-glucan) or 45 days with a diet containing 0.1% of ß-glucan (MacroGard®). We evaluated the growth performance at the end of the ß-glucan feeding trial and the innate immune function immediately after the feeding trial and 7 and 14 days post-feeding trial. In addition, at day 10 post-feeding trial, we assessed the tilapia's resistance against a bacterial infection. No significant differences were observed in growth performance between the groups; however, fish fed with ß-glucan for 30 and 45 days had higher (approx. 8%) relative weight gain compared to the control. Regardless of the administration period, fish fed with ß-glucan had higher innate immune responses immediately after the feeding trial such as lysozyme activity in plasma, liver and intestine and respiratory burst compared to the control, and in general these differences were gradually reduced over the withdrawal period (up to 14 days). No differences were observed in the plasma hemolytic activity of the complement or myeloperoxidase activity in plasma or intestine. Moreover, fish from the control group had early mortalities (2 vs. 4-5 days post-infection, respectively) and a lower survival rate (60 vs. 80%, respectively) compared to fish fed with ß-glucan for 15 or 30 days, and, interestingly, fish fed for 45 days with ß-glucan had no mortality. This study indicates that regardless of the administration period (i.e., 15 up to 45 days), the ß-glucan improved the innate immune responses and the tilapia's resistance to disease, and this protection could be observed up to 10 days post-feeding trial, adding in vivo evidence that ß-glucan may contribute to a trained innate immunity. Additionally, we showed that a longer period of administration did not cause immunosuppression as previously hypothesized but promoted further growth and immune performance. These findings are relevant to the aquaculture industry and demonstrate that a longer ß-glucan feeding protocol may be considered to achieve better results.

Preclinical safety and immunogenicity of Streptococcus pyogenes (Strep A) peptide vaccines.

We have developed two candidate vaccines to protect against multiple strains of Strep A infections. The candidates are combinatorial synthetic peptide vaccines composed of a M protein epitope (J8 or p*17) and a non-M protein epitope (K4S2). To enhance immunogenicity, each peptide is conjugated to the carrier protein CRM197 (CRM) and formulated with aluminium hydroxide adjuvant Alhydrogel (Alum) to make the final vaccines, J8-CRM + K4S2-CRM/Alum and p*17-CRM + K4S2-CRM/Alum. The safety and toxicity of each vaccine was assessed. Sprague Dawley rats were administered three intramuscular doses, over a six-week study with a 4-week recovery period. A control group received CRM only formulated with Alum (CRM/Alum). There was no evidence of systemic toxicity in the rats administered either vaccine. There was an associated increase in white blood cell, lymphocyte and monocyte counts, increased adrenal gland weights, adrenocortical hypertrophy, and increased severity of granulomatous inflammation at the sites of injection and the associated inguinal lymph nodes. These changes were considered non-adverse. All rats administered vaccine developed a robust and sustained immunological response. The absence of clinical toxicity and the development of an immunological response in the rats suggests that the vaccines are safe for use in a phase 1 clinical trial in healthy humans.

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.

Dietary inclusion of chestnut (Castanea sativa) polyphenols to Nile tilapia reared in biofloc technology: Impacts on growth, immunity, and disease resistance against Streptococcus agalactiae.

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.

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.

Lancefield Whole Blood Killing Assay to Evaluate Vaccine Efficacy.

While the Lancefield whole blood killing assay is named after the renowned streptococcal researcher Rebecca Lancefield, the protocol was first described by Todd in 1927 (Br J Exp Pathol 8:1-5, 1927). Initially, the assay was used to identify novel Group A Streptococcal (GAS) serotypes through the supplementation of non-immune human blood (often from infants) with type-specific antisera prepared in rabbits (Lancefield, J Exp Med 106:525-544, 1957; Maxted, Br J Exp Pathol 37:415-422, 1956) and to demonstrate the impressive longevity of type-specific immunity in patients following invasive GAS infection (Lancefield, J Exp Med 110:271-292, 1959). The modern assay is routinely used to screen defined GAS mutants (Wessels, Bronze, Proc Natl Acad Sci U S A 91:12238-12242, 1994; Zinkernagel et al., Cell Host Microbe 4:170-178, 2008) or transposon libraries (Le Breton et al., Infect Immun 81:862-875, 2013) for enhanced susceptibility to opsonophagocytic killing or to screen vaccine antisera (Salehi et al., mSphere 3:e00617-e00618, 2018) or other serological preparations (Reglinski et al., Sci Rep 5:15825, 2015) for anti-streptococcal activity.

Immunostimulatory effects of Sarcodia suiae water extracts on Nile tilapia Oreochromis niloticus and its resistance against Streptococcus agalactiae.

In this study, water extracts of the red seaweed Sarcodia suiae were obtained using solid-liquid extraction (SLE) or pressurized liquid extraction (PLE) methods. The extracts were used to investigate immunostimulatory activity by measuring the phagocytic activity of Nile tilapia (Oreochromis niloticus) hepatic and splenic macrophages and the tilapia head kidney (THK) cell line, and modulation of immune-related genes in primary head kidney (HK) cells and THK cells. At 10 µg/ml, both extracts promoted the proliferation of hepatic and splenic macrophages. Expression levels of proinflammatory cytokines (IL-1ß and IL-8), antimicrobial peptides (TP2 and TP4), and pattern recognition receptors (TLR5) were elevated in SLE extracts-treated primary HK leukocytes. Similarly, IL-1ß, IL-8, and TNFα expression was also induced by SLE extract in THK cells. Phagocytic activity in primary HK cells and THK cells was induced by SLE extract 12 h and 24 h post-stimulation, while PLE extract only induced phagocytic activity in THK cells at early time points. SLE extract (100 µg/g body weight) increased the expression of IL-1ß, IL-8, TNFα, TP2, TP4, TLR2 and TLR5 in the spleen and immunoprotective efficiency against Streptococcus agalactiae infection. Taken together, these results show that S. suiae can differentially stimulate the immune response of tilapia in vitro and in vivo and could potentially be used as an immunomodulator in tilapia culture.

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.

Streptococcal H2O2 inhibits IgE-triggered degranulation of RBL-2H3 mast cell/basophil cell line by inducing cell death.

Mast cells and basophils are central players in allergic reactions triggered by immunoglobulin E (IgE). They have intracellular granules containing allergic mediators (e.g., histamine, serotonin, inflammatory cytokines, proteases and ß-hexosaminidase), and stimulation by IgE-allergen complex leads to the release of such allergic mediators from the granules, that is, degranulation. Mast cells are residents of mucosal surfaces, including those of nasal and oral cavities, and play an important role in the innate defense system. Members of the mitis group streptococci such as Streptococcus oralis, are primary colonizers of the human oral cavity. They produce hydrogen peroxide (H2O2) as a by-product of sugar metabolism. In this study, we investigated the effects of streptococcal infection on RBL-2H3 mast cell/basophil cell line. Infection by oral streptococci did not induce degranulation of the cells. Stimulation of the RBL-2H3 cells with anti-dinitrophenol (DNP) IgE and DNP-conjugated human serum albumin triggers degranulation with the release of ß-hexosaminidase. We found that S. oralis and other mitis group streptococci inhibited the IgE-triggered degranulation of RBL-2H3 cells. Since mitis group streptococci produce H2O2, we examined the effect of S. oralis mutant strain deficient in producing H2O2, and found that they lost the ability to suppress the degranulation. Moreover, H2O2 alone inhibited the IgE-induced degranulation. Subsequent analysis suggested that the inhibition of degranulation was related to the cytotoxicity of streptococcal H2O2. Activated RBL-2H3 cells produce interleukin-4 (IL-4); however, IL-4 production was not induced by streptococcal H2O2. Furthermore, an in vivo study using the murine pollen-induced allergic rhinitis model suggested that the streptococcal H2O2 reduces nasal allergic reaction. These findings reveal that H2O2 produced by oral mitis group streptococci inhibits IgE-stimulated degranulation by inducing cell death. Consequently, streptococcal H2O2 can be considered to modulate the allergic reaction in mucosal surfaces.

Modulation of mucosal parameters, innate immunity, growth and resistance against Streptococcus agalactiae by enrichment of Nile tilapia (Oreochromis niloticus) diet with Leucas aspera.

The present study aimed at evaluating the possible effects of Leucas aspera as immunostimulant on mucosal and serum immunity, as well as on growth and resistance against Streptococcus agalactiae infection in Nile tilapia (Oreochromis niloticus) fingerlings. In a 45 days trial, fish were fed experimental diets containing L. aspera 0 g kg-1 (Diet 1- control), 1 g kg-1 (Diet 2), 2 g kg-1(Diet 3), 4 g kg-1 (Diet 4) and 8 g kg-1 (Diet 5). The results revealed a significant increase in the specific growth rate (SGR), weight gain (WG), and final weight (FW) in fish fed diet 3 (2 g kg-1) of L. aspera compared to the control and other supplemented groups (P < 0.05). Also, feeding on diet 3 (2 g kg-1) of L. aspera enriched diet significantly (P < 0.05) increased lysozyme activities in the serum and mucus, serum peroxidase and phagocytosis activity. However, significant (P < 0.05) increase in mucus peroxidase activity was reported in fish fed diet 4 (4 g kg-1) and diet 5 (8 g kg-1) of L. aspera, whereas significantly higher (P < 0.05) alternative complement activity was reported in fish fed diet 2 (1 g kg-1) of L. aspera. At the end of the experiment, nine fish per replication were selected for a challenge test against S. agalactiae. The dietary supplementation of L. aspera significantly reduced the mortality rate and increased the resistance of Nile tilapia following by challenge with S. agalactiae. The highest post challenge survival of 100% was observed in tilapia fed diet 5 (8 g kg-1) following by 92.6% of RPS in fish fed diet 4 (4 g kg-1) and 88.9% in diet 3 (2 g kg-1), 77.8% in diet 2 (1 g kg-1) and 74.1% in diet 1(0 g kg-1).

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.

Effects of elephant's foot (Elephantopus scaber) extract on growth performance, immune response, and disease resistance of nile tilapia (Oreochromis niloticus) fingerlings.

Medicinal plant has been applied as an alternative strategy for antibiotics and chemotherapeutics for controlling the outbreak of diseases in tilapia farming. In this study, five doses of Elephantopus scaber extract (ESE) were added to the basal diet at 0, 2.5, 5, 10, and 20 g kg-1 feed of Nile tilapia fingerlings (13.92 ±â€¯0.06 g initial weight) in triplicate. After 4- and 8- weeks post-feeding, fish were sampled to determine the effects of the ESE supplemented on fish's growth performance, humoral, and skin mucus immune response. After 8 weeks post-feeding, a challenge test against Streptococcus agalactiae was carried out using 10 fish from each tank. Fish fed ESE showed significantly increased serum lysozyme (SL), serum peroxidase (SP), alternative complement (ACH50), phagocytosis (PI), and respiratory burst (RB) compared to the control group (P < 0.05). The skin mucus lysozyme (SMLA) and skin peroxidase (SMPA) were stimulated in fish fed ESE diets. Dietary inclusion of ESE significantly (P < 0.05) promoted final body weight (FW), weight gain (WG), and specific growth rate (SGR); while a reduction in feed conversion ratio (FCR) was observed in fish fed 5 g kg-1 ESE, after 8 weeks post-feeding. The challenge study indicated that the relative percent survival (RSP) was 38.10%, 76.19%, 66.67%, and 47.62% in Diet 2, Diet 3, Diet 4, and Diet 5, respectively. Among the supplemented groups, dietary of 5 g kg-1 ESE showed significantly higher RPS and the highest resistance to S. agalactiae in comparison with other groups. In conclusion, supplementation of ESE (5 g kg-1) enhanced the humoral and mucosal immunity, promoted growth performance, and improved disease resistance of Nile tilapia against Streptococcus agalactiae.