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.

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

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

Curcumin loaded liposome formulation: Enhanced efficacy on performance, flesh quality, immune response with defense against Streptococcus agalactiae in Nile tilapia (Orechromis niloticus).

Application of novel trend comprising antioxidant phytogenics is aiming to minimize the stress related factors and associated diseases in intensive fish culturing. Today, the concept of exploiting and protecting natural antioxidants represents a paradigm shift for the aqua feed industry. Therefore, our principal goal targeting liposome as a novel nanocarrier for curcumin is directed to attain superior performance, fillet antioxidant stability and bacterial resistance in Nile tilapia. A total of 500 Nile tilapia fingerlings (average body weight, 10.27 ± 0.10 g) assigned into five experimental groups in 25 glass aquaria of 120 L capacity at the density 20 fish/aquaria. The experimental groups were supplemented with varying doses of liposomal curcumin-NPs, LipoCur-NPs (0, 5, 15, 25 and 35 mg/kg diet) were reared for 12 weeks and later Streptococcus agalactiae (S. agalactiae) challenged model was performed. Inclusion of LipoCur-NPs (25 and 35 mg/kg diet) had the most prominent impact on Nile tilapia growth rate and feed conversion ratio. The immune boosting outcomes post supplementing 35 mg/kg diet of LipoCur-NPs were evidenced by higher myeloperoxidase, lysozyme and total immunoglobulin levels. Even after 4 weeks frozen storage, LipoCur-NPs at the dose of 35 mg/kg diet prominently increased (P < 0.05) the fillet scavenging capability for free radicals (1,1-diphenyl-2-picrylhydrazyl and 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) with an inverse reduction in lipid peroxidation biomarker (malondialdehyde). Notably, upregulation of GSH-Px, CAT, and SOD genes in fillet of 35 mg/kg LipoCur-NPs fed fish coordinated with higher T-AOC and lower oxidative markers (ROS and H2O2). Post S. agalactiae challenge, higher supplementation levels of LipoCur-NPs (35 mg/kg diet) greatly attenuated the expression of its vital virulence genes (cfb, fbsA and cpsA) with higher expression of Igm, CXC-chemokine and MHC genes. Concordantly, downregulation of inflammatory markers (IL-1ß, TNF-α and IL-8) and upregulation of anti-inflammatory ones (IL-10 and TGF-ß) were remarkably documented. Based on these findings, the innovative curcumin loaded liposome was considered a novel multitargeting alternative not only playing an imperative role in Nile tilapia growth promotion and fillet stability upon storage, but also protecting efficiently against S. agalactiae.

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

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

Effects of Elephantopus scaber extract on growth, proximate composition, immunity, intestinal microbiota and resistance of the GIFT strain of Nile tilapia Oreochromis niloticus to Streptococcus agalactiae.

This study aimed to investigate the effects of Elephantopus scaber extract on the GIFT (genetic improvement of farmed tilapia) strain of Nile tilapia Oreochromis niloticus. A total of 800 tilapia with an initial body weight of 1.34 ± 0.09 g each were randomly divided into five groups. The tilapia in the control group (E0 group) were fed on a basal diet only. Meanwhile, tilapia in the four experimental groups were fed on a basal diet supplemented with 1 g/kg (E1 group), 3 g/kg (E2 group), 5 g/kg (E3 group), and 7 g/kg (E4 group) of E. scaber extract for 10 weeks. Results showed that the survival rate was higher in the experimental groups than in the control group. Compared with the control group, some growth parameters (FW, WGR, SGR, VSI, and HSI) were significantly improved in the E1 group and E2 group. The crude lipid content in the dorsal muscle and liver was lower in the E1 group than in the control group. After E. scaber extract supplementation, activities of immunity-related enzymes (ACP, AKP, T-AOC, SOD, CAT, GSH-Px and LZM) in plasma, liver, spleen and head kidney, and expressions of immunity-related genes (IL-1ß, IFN-γ, TNF-α, and CCL-3) in liver, spleen and head kidney showed various degrees of improvement, while MDA content and Hsp70 expression level were decreased. The survival rate of tilapia increased in all the supplementation groups after Streptococcus agalactiae treatment. E. scaber extract addition changed the species composition, abundance, and diversity of intestinal microbiota in tilapia. These results demonstrate that E. scaber extract supplementation in diet can improve the growth, immunity, and disease resistance of GIFT against S. agalactiae. E. scaber extract supplementation can also change intestinal microbiota and reduce crude lipid content in dorsal muscle and liver. The above indicators show that the optimal dose of E. scaber extract for GIFT is 1 g/kg.

Garlic: An Alternative Treatment for Group B Streptococcus.

Prenatal screening in pregnant women between 35 and 37 weeks of gestation and intrapartum antibiotic prophylaxis has successfully reduced the incidence of neonatal morbidity and mortality related to Streptococcus agalactiae. However, the contamination rates of newborns are still considerable. In traditional and folk medicines, it has been observed that garlic has been effective in treating S. agalactiae infection. The aim of this study was to isolate and identify the active compounds from garlic that have antimicrobial activity against S. agalactiae. In order to do this, SP80 (Sep-Pak 80%) obtained from crude garlic extract (CGE) was fractionated by reverse-phase ultrafast liquid chromatography with UV (RP-UFLC-UV) using a Shim-pack PREP-ODS column. All fractions obtained were tested using a microbial growth inhibition test against the S. agalactiae strain (ATCC 12386). Five clinical isolates were used to confirm the action of the fractions with antimicrobial activity, and the bacterial growth curve was determined. Identification of the antimicrobial compounds was carried out through liquid chromatography coupled with mass spectrometry (LC/MS) and nuclear magnetic resonance (NMR). The active compounds found to exhibit antimicrobial activity were Ƴ-glutamyl-S-allyl-cysteine (fraction 18), Ƴ-glutamyl-phenylalanine (fraction 20), and the two stereoisomers (E and Z) of ajoene (fraction 42). The MICs of these fractions were 5.41 mg/ml, 4.60 mg/ml, and 0.16 mg/ml, respectively, and they inhibited the growth of the clinical isolates tested. Antimicrobial compounds from garlic may be a promising source in the search for new drugs against S. agalactiae. IMPORTANCE Invasive disease due to group B streptococcal (GBS) infection results in a wide spectrum of clinical disease in neonates. Maternal colonization by GBS is the primary risk factor for disease. The strategy recommended by the Centers for Disease Control to reduce neonatal GBS infection is the culture-based screening of all pregnant women at 35 to 37 weeks of gestation and intrapartum antibiotic prophylaxis (IAP). However, indiscriminate use of antibiotics favors the selection and spread of resistant bacteria. The global scenario of antibacterial resistance has been of great concern for public health, and natural products can be a source of new substances to help us grapple with this problem.

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.

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.

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.

Effects of Assam tea extract on growth, skin mucus, serum immunity and disease resistance of Nile tilapia (Oreochromis niloticus) against Streptococcus agalactiae.

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

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.

Traditional Chinese Medicine Enhances Growth, Immune Response, and Resistance to Streptococcus agalactiae in Nile Tilapia.

Herbal supplements are suitable for improving fish health and combating diseases in fish culture. However, the mechanism of action of many herbal mixtures remains unclear. This study was conducted to evaluate the effects of traditional Chinese medicine (TCM; a mix of Astragalus membranaceus, Angelica sinensis, and Crataegus hupehensis at a ratio of 1:1:1 on a weight basis) on growth, immune response, and disease resistance in Nile tilapia Oreochromis niloticus. Experimental fish (mean ± SE weight = 57 ± 1 g) were divided into two groups: a control group and a TCM (10 g/kg) group. After 4 weeks of the experimental trial, a significant increase in weight gain and specific growth rate and a lower feed conversion ratio were observed in fish fed a TCM-supplemented diet compared with control fish. Similarly, the immune response of Nile Tilapia in the TCM group showed enhanced lysozyme, superoxide dismutase, catalase, and immunoglobulin levels compared with the control fish. In comparison with the control, fish fed TCM showed significant up-regulation of ß-defensin, lysozyme, heat shock protein 70, superoxide dismutase, and catalase genes in the intestine and head-kidney tissues. After a Streptococcus agalactiae challenge, survival of Nile Tilapia in the TCM group was 70% compared with 35% in the control. These results indicate that the TCM mixture in this study can elevate the immune response and disease resistance of Nile Tilapia.

Potential Application of Bacteriophages in Enrichment Culture for Improved Prenatal Streptococcus agalactiae Screening.

Vertical transmission of Streptococcus agalactiae can cause neonatal infections. A culture test in the late stage of pregnancy is used to screen for the presence of maternal S. agalactiae for intrapartum antibiotic prophylaxis. For the test, a vaginal⁻rectal sample is recommended to be enriched, followed by bacterial identification. In some cases, Enterococcus faecalis overgrows in the enrichment culture. Consequently, the identification test yields false-negative results. Bacteriophages (phages) can be used as antimicrobial materials. Here, we explored the feasibility of using phages to minimize false-negative results in an experimental setting. Phage mixture was prepared using three phages that specifically infect E. faecalis: phiEF24C, phiEF17H, and phiM1EF22. The mixture inhibited the growth of 86.7% (26/30) of vaginal E. faecalis strains. The simple coculture of E. faecalis and S. agalactiae was used as an experimental enrichment model. Phage mixture treatment led to suppression of E. faecalis growth and facilitation of S. agalactiae growth. In addition, testing several sets of S. agalactiae and E. faecalis strains, the treatment with phage mixture in the enrichment improved S. agalactiae detection on chromogenic agar. Our results suggest that the phage mixture can be usefully employed in the S. agalactiae culture test to increase test accuracy.

Sylimarin as hepatic protector and immunomodulator in Nile tilapia during Streptococcus agalactiae infection.

This study investigated the use of silymarin, an extract obtained from the milk thistle (Silybum marianum) and its effects as a possible hepatoprotector in Nile tilapia (Oreochromis niloticus). Silymarin was used as feed additive to the diet at a concentration of 0.1% (1 kg per ton of dry ration) with the commercial product named Di-Heptarine S® (16% silymarin phosphatide). A total of 90 juvenile tilapia with approximately 45 days old and mean weight of 0.72 ±â€¯0.04 g were distributed in two groups, one fed with a diet with the hepatoprotector and the other without the additive. At the end of the assay (55 days after feeding), samples of blood were collected for hematological, immunological, histological (liver, spleen and intestine) and enzymatic analysis such as superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST). After 55 days all fish were challenged with Streptococcus agalactiae serotype Ib to verify the sylimarin effects on the immunological parameters and its protection effect while challenged. During the challenge period another biological material sample was collected for hematological, immunological and histopathological analysis (liver, spleen and intestine). Before the challenge, an increase on the count of thrombocyte was found in the supplemented fish. In the liver, dilation of the sinusoids was observed in unsupplemented fish while supplemented fish the alteration was less severe. No significant alteration was found in SOD, CAT and GST between the groups. Histological changes after the challenge were provoked by bacterial toxins as a result of inflammatory processes. Periacinar degeneration was less intense in unsupplemented fish when compared to supplemented fish. On the other hand, eosinophilic and lymphocytic infiltrate did occur in unsupplemented fish differently from supplemented fish which did not show the alteration. The survival was 28% higher in silymarin supplemented fish when compared to unsupplemented fish that presented no survival. Silymarin supplementation in the diet provided a hepatoprotective and immunomodulatory effect on Nile tilapia.

Different ß-glucans improve the growth performance and bacterial resistance in Nile tilapia.

The role of glucan as a biologically active immunomodulator has been well documented for more than 40 years. However, the wide diversity of ß-glucan forms and the extraction process has implications for the benefits of these compounds. Biorigin developed two samples of ß-glucans using different biotechnological processes. Thus, in the present study, we fed Nile tilapia (Oreochromis niloticus) diets containing these two ß-glucan molecules (BG01 and BG02) for 30 days prior to bacterial infection with Streptococcus agalactiae. The results showed that the different ß-glucan samples exhibited biologically differently behaviors, but both increased the resistance against bacterial infection. Specifically, BG01 increased immunostimulation, while BG02 improved growth performance. In summary, these findings confirm the benefits of ß-glucans in aquaculture and also provide further evidence of the growth promotion of these compounds.

Effects of Cordyceps militaris spent mushroom substrate on mucosal and serum immune parameters, disease resistance and growth performance of Nile tilapia, (Oreochromis niloticus).

The aim of present study was determination effects of dietary administration of C. militaris spent mushroom substrate (SMS) on mucosal and serum immune parameters, disease resistance, and growth performance of Nile tilapia (Oreochromis niloticus). Two hundred twenty five fish of similar weight (37.28 ± 0.10 g) were assigned to the following diets [0 (T1- Control), 5 (T2), 10 (T3), 20 (T4) and 40 g kg-1 (T5) SMS]. After 60 days of feeding trial, growth performance, skin mucus lysozyme and peroxidase activities as well as serum innate immune were measured. In addition, survival rate and innate immune responses were calculated after challenge test (15 days) against Streptococcus agalactiae. The results revealed that regardless of inclusion levels, feeding Nile tilapia with SMS supplemented diets significantly increased skin mucus lysozyme and peroxidase activities as well as serum immune parameters (SL, ACH50, PI, RB, and RB) compared control group (P < 0.05). The highest increment of immune parameters was observed in fish fed 10 g kg-1 SMS which was significantly higher than other treatments (P < 0.05). Also, the relative percent survival (RSP) in T2, T3, T4, and T5 was 61.11%, 88.89%, 66.67, and 55.56%, respectively. Among the supplemented groups, fish fed 10 g kg-1 SMS showed significant higher RPS and resistance to S. agalactiae than other groups (P < 0.05). Regarding the growth performance, SGR, WG, FW, and FCR were remarkably improved (P < 0.05) in SMS groups; the highest improvement observed in 10 g kg-1 SMS treatment. According to these finding, administration of 10 g kg-1 SMS is suggested in tilapia to improve growth performance and health status.

The effects of dietary kefir and low molecular weight sodium alginate on serum immune parameters, resistance against Streptococcus agalactiae and growth performance in Nile tilapia (Oreochromis niloticus).

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

Combined administration of low molecular weight sodium alginate boosted immunomodulatory, disease resistance and growth enhancing effects of Lactobacillus plantarum in Nile tilapia (Oreochromis niloticus).

The present study investigates the effects of combined or singular administration of low molecular weight sodium alginate (LWMSA) and Lactobacillus plantarum on innate immune response, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus). Three hundred and twenty fish were supplied and randomly stocked in sixteen glass tanks (150 L) assigned to four treatments as follows: 0 g kg-1 LMWSA (Control, Diet 1), 10 g kg-1 LMWSA (Diet 2), 108 CFU g-1L. plantarum (Diet 3), and 10 g kg-1 LMWSA + 108 CFU g-1L. plantarum (Diet 4). Following 30 and 60 days of the feeding trial, serum lysozyme, phagocytosis, respiratory burst and alternative complement activities as well as growth performance parameters (specific growth rate, feed conversion ratio) were measured. Serum lysozyme, phagocytosis, respiratory burst, and alternative complement activities of fish were significantly stimulated by both LMWSA and L. plantarum diets, however, the highest innate immune response were observed in fish fed synbiotic diet. At the end of the experiment, eight fish per replication were randomly selected for a challenge test against Streptococcus agalactiae. The survival rate of the fish fed supplemented diets was significantly greater than the control treatment and the highest post challenge survival rate was observed in synbiotic diet. Furthermore, SGR and FCR were significantly improved in fish fed supplemented diets after 60 days and the highest growth performance was observed in fish fed synbiotic diet. These results suggest combined LMWSA and L. plantarum can be considered as a promising immunostimulant and growth enhancer in Nile tilapia diet.

Effects of low molecular weight sodium alginate on growth performance, immunity, and disease resistance of tilapia, Oreochromis niloticus.

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