Effects of dietary nanoliposome-coated astaxanthin on haematological parameters, immune responses and the antioxidant status of rainbow trout (Oncorhynchus mykiss).

BACKGROUND: Astaxanthin is the most prevalent carotenoid in the marine environment and is widely used as an additive in formulated aquafeeds. OBJECTIVES: A 60-day feeding trial was conducted to consider the effect of dietary nanoliposome-coated astaxanthin (NA) on haematological parameters, serum antioxidant activities and immune responses of rainbow trout, Oncorhynchus mykiss. METHODS: A total of 450 healthy fish weighing 31.00 ± 2.09 g were randomly assigned in triplicate (30 fish per replicate) to 5 dietary treatments: 0 (control), 25.00, 50.00, 75.00, and 100.00 mg kg-1 NA. RESULTS: Fish fed the diet supplemented with 50.00 mg kg-1 NA exhibited the highest values of red blood cells, white blood cells, haemoglobin and haematocrit of 1.64 ± 0.01 × 106 mm-3, 5.54 ± 0.21 × 103 mm-3, 8.73 ± 0.24 g dL-1 and 46.67% ± 0.88%, respectively, which were significantly higher than those fed the basal diet (p < 0.05). The lowest and highest percentages of lymphocytes (67.67% ± 0.33%) and neutrophils (27.33% ± 1.20%) were also obtained in fish fed 50.00 mg kg-1 NA compared to those fed the basal diet (p < 0.05). Fish receiving diet supplemented with 50.00 mg kg-1 NA revealed the highest serum activity in superoxide dismutase, catalase, glutathione peroxidase, lysozyme and alternative complement and the lowest level of total cholesterol, cortisol, aspartate aminotransferase and alanine aminotransferase than fish receiving the basal diet (p < 0.05). Serum immunoglobulin (Ig) and ACH50 contents significantly increased with increasing dietary NA supplementation to the highest values of 43.17 ± 1.46 and 293.33 ± 2.03 U mL-1, respectively, in fish fed diet supplemented with 50 mg kg-1 NA (p < 0.05). CONCLUSIONS: Supplementation of NA in rainbow trout diet at 50 mg kg-1 exhibited a positive effect on haematological parameters, antioxidant capacity and immune responses. Administration of such dosage can enhance rainbow trout immune responses against unfavourable or stressful conditions, for example disease outbreaks, hypoxic condition, thermal stress and sudden osmotic fluctuations, which usually happen in an intensive culture system.

Streptococcosis a Re-Emerging Disease in Aquaculture: Significance and Phytotherapy.

Streptococcosis, particularly that caused by S. iniae and S. agalactiae, is a major re-emerging bacterial disease seriously affecting the global sustainability of aquaculture development. Despite a wide spread of the disease in aquaculture, few studies have been directed at assessing the in vitro antagonistic activity and in vivo efficacy of medicinal herbs and other plants against streptococcal agents. Most in vitro studies of plant extractives against S. iniae and S. agalactiae have found antibacterial activity, but essential oils, especially those containing eugenol, carvacrol or thymol, are more effective. Although essential oils have shown better anti-streptococcal activity in in vitro assays, in vivo bioassays require more attention. The extracts examined under in vivo conditions show moderate efficacy, increasing the survival rate of infected fish, probably through the enhancement of immunity before challenge bioassays. The available data, however, lack dosage optimization, toxicity and bioavailability assays of a specific plant or its bioactive compound in fish organs; hence, it is difficult to judge the validation of clinical efficacy for the prevention or treatment of fish streptococcosis. Despite the known bioactive compounds of many tested plants, few data are available on their mode of action towards streptococcal agents. This review addresses the efficacy of medicinal plants to fish streptococcosis and discusses the current gaps.

Immunomulation effect of alginic acid and chitooligosaccharides in silver carp (Hypophthalmichthys molitrix).

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

Study on antioxidant potential, immunological response, and inflammatory cytokines induction of glycyrrhizic acid (GA) in silver carp against vibriosis.

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

Lactococcosis a Re-Emerging Disease in Aquaculture: Disease Significant and Phytotherapy.

Lactococcosis, particularly that caused by Lactococcus garvieae, is a major re-emerging bacterial disease seriously affecting the sustainability of aquaculture industry. Medicinal herbs and plants do not have very much in vitro antagonism and in vivo disease resistance towards lactococcosis agents in aquaculture. Most in vitro studies with herbal extractives were performed against L. garvieae with no strong antibacterial activity, but essential oils, especially those that contain thymol or carvacrol, are more effective. The differences exhibited by the bacteriostatic and bactericidal functions for a specific extractive in different studies could be due to different bacterial strains or parts of chemotypes of the same plant. Despite essential oils being shown to have the best anti-L. garvieae activity in in vitro assays, the in vivo bioassays required further study. The extracts tested under in vivo conditions presented moderate efficacy, causing a decrease in mortality in infected animals, probably because they improved immune parameters before challenging tests. This review addressed the efficacy of medicinal herbs to lactococcosis and discussed the presented gaps.

Effects of dietary chitosan and nano-chitosan loaded clinoptilolite on growth and immune responses of rainbow trout (Oncorhynchus mykiss).

In this study, rainbow trout Oncorhynchus mykiss weighing 27.75 ± 0.34 g were orally subjected to eight experimental diets each in three replicates containing varying amounts of chitosan and nano-chitosan (0.05, 0.5 and 5 g kg-1) loaded in clinoptilolite (14.28 g kg-1) for 70 days; and the growth and immune responses were evaluated. Results showed that growth parameters in fish fed diets chit + clin2, chit + clin3, nchit + clin1, nchit + clin2 and nchit + clin3 were significantly higher than in fish fed the control diet. All feeds, except chit + clin3, and nchit + clin3, significantly increased the total protein level. Feeds containing chit + clin2, nchit + clin1, and nchit + clin2 significantly elevated serum lysozyme activity compared with the control group. All treatments, except chit + clin3, and nchit + clin3 exhibited higher serum immunoglobulin (Ig) level than control one. In contrast, diet nchit + clin1 significantly unregulated the expression of Ig M gene in fish head-kidney compared to other groups. Additionally, all feeds, except clinoptilolite, and nchit + clin3, significantly improved the serum complement activity. Diets chit + clin2, nchit + clin1, and nchit + clin2 also significantly elevated antibacterial activity against Yersinia ruckeri compared with the control diet. Expression of inducible nitric oxide synthase (iNOS) gene in fish fed diets clinoptilolite, chit + clin1, chit + clin3, nchit + clin1, nchit + clin2, and nchit + clin3 was significantly higher than the control diet. All diets, except clinoptilolite, increased IL-1ß gene expression compared to the control group. Present results suggest that diets supplemented with nchit + clin, especially at 0.05 g kg-1 nano-chitosan inclusion, could improve growth performance and immune parameters of rainbow trout.

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.

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.

Protective effect of zinc on related parameters to bone metabolism in common carp fish (Cyprinus carpio L.) intoxified with cadmium.

The short term effects of waterborne cadmium (Cd(+2)) on the levels of serum parameters related to bone metabolism including calcium (Ca), inorganic phosphorus (P(i)) and alkaline phosphatase (ALP) in common carp fish (Cyprinus carpio L.) were studied. Fish were treated with varying concentrations of Cd(+2) (0.22, 1.1 and 2.2 mg l(-1)) daily for 14 days. The results obtained show that serum P(i) and ALP concentrations were elevated by increasing Cd(+2) concentration in water containing fish whereas serum Ca level was decreased. At the same time, the protective role of waterborne zinc (Zn(+2), 1 mg l(-1)) on the same parameters was also investigated. Results showing that daily treatment of fish with Zn(+2), increased the concentrations of Ca and ALP in serum by 2.07 and 1.86 fold and decreased serum P(i) level by 57.7% in comparison with Cd(+2) treatment. The combination of Cd(+2) and Zn(+2) on the same parameters was studied next. There was a significant (P < 0.05) elevation in serum Ca and ALP levels in comparison with Cd(+2) treatment. Decreasing in serum P(i) level was not significant in comparison with Cd(+2) treatments. The protective effect of Zn(+2) on Cd(+2) disturbances in serum parameters related to bone metabolism in this manuscript has been also discussed.