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

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

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.

The effects singular or combined administration of fermentable fiber and probiotic on mucosal immune parameters, digestive enzyme activity, gut microbiota and growth performance of Caspian white fish (Rutilus frisii kutum) fingerlings.

The aim of the present study was to investigate the effects of single or combined administration of dietary fermentable fiber (Vitacel®) and probiotic PrimaLac® on mucosal immune parameters, digestive enzyme activity, gut microbiota and growth performance of Caspian white fish (Rutilus frisii kutum) fingerlings. Fish were transferred to laboratory, acclimatized for two weeks and then fish (0.56 ±â€¯0.026 g) were allocated into 12 tanks (30 fish per tank). Triplicate groups were fed a basal diet (Control) or basal diet supplemented with fermentable fiber [Vitacel®] (FF), probiotic [PrimaLac®] (P) and combined fermentable fiber and probiotic (FF + P). At the end of feeding trial, growth performance and feed utilization parameters were significantly (P < 0.05) improved in FF, P and FF + P treatments compared control group. Evaluation of digestive enzyme activity revealed significant (P < 0.05) increase of lipase activity in fish fed supplemented diet. However, amylase, protease and alkaline phosphatase were significantly higher (P < 0.05) only in P and FF + P treatments. Furthermore, total autochthonous intestinal microbiota and autochthonous LAB levels significantly increased in fish fed supplemented diet (P < 0.05). Also, inclusion of FF, P and FF + P in Caspian white fish diet remarkably increased skin mucus immune parameters compared control group (P < 0.05). These results indicate that singular or combined administration of FF and P can be considered as a beneficial dietary supplement for early stages of Caspian white fish (Rutilus fresii kutumn) culture.

Dietary Lactobacillus acidophilus modulated skin mucus protein profile, immune and appetite genes expression in gold fish (Carassius auratus gibelio).

The objective of the present study was to investigate the effect of dietary Lactobacillus acidophilus on skin mucus protein pattern, immune and appetite related genes expression as well as growth performance in gold fish (Carassius auratus gibelio). Three hundred healthy gold fish (2.5 ± 0.05) juveniles were randomly distributed in 12 glass aquariums (400-L; 25 fish per aquaria) and fed experimental diets contain different levels of L. acidophilus (0, 1.5 × 108, 3 × 108 and 6 × 108) for 8 weeks. SDS-PAGE analysis of skin mucus protein profile at the end of the feeding trial revealed differences in protein profile of probiotic fed fish and control group; even three new bands were observed in L. acidophilus treated groups. Furthermore, fish fed 6 × 108 CFU g-1 supplemented diet showed up-regulation of both TNF-1α and TNF-2α gene expression (P < 0.05). Evaluation of appetite related gene expression showed down-regulation of ghrelin in probiotic fed fish compared those of control treatment (P < 0.05). However, administration of different levels of L. acidophilus had no significant effects on growth performance (P > 0.05). These results demonstrated that while no beneficial effects on growth performance, dietary L. acidophilus affects immune and appetite related genes expression as well as skin mucus protein profile.

Immune related transcriptional responses and performance of Litopenaeus vannamei post-larvae fed on dietary probiotic PrimaLac(®).

The present study investigated the effects of various levels of multi-strain probiotic on the immune related gene expression, digestive enzyme activity, growth performance, body chemical composition and survival of Litopenaeus vannamei post-larvae. After transferring post-larvae of L. vannamei to indoor conditions and subsequent acclimation to laboratory condition for 14 days, the shrimp were fed multi-strain probiotic at four different doses of 0, 0.25, 0.5 and 1.0 g kg(-1) for eight weeks. Shrimp fed 0.5 and 1.0 g kg(-1) probiotic PrimaLac(®) exhibited significantly (p < 0.05) higher weight gain, specific growth rate, body crude protein as well as lower FCR compared to other groups. Feeding on 0.5 and 1 g kg(-1) dietary multi-strain probiotic significantly (p < 0.05) increased the level of body crude protein. Oral administration of 0.5 and 1.0 g kg(-1) multi-strain probiotic significantly (p < 0.05) decreased body crude lipid and body moisture respectively. 30 days after feeding, protease, amylase and lipase activity increased in groups fed 0.5 and 1.0 g kg(-1) probiotic PrimaLac(®). However, on the 60th day, specific protease and amylase activity in all treatment groups were significantly higher than control group (p < 0.05) but lipase activity was higher (p < 0.05) in groups fed 0.5 and 1.0 g kg(-1) multi-strain probiotic. Oral administration of 1.0 g kg(-1) probiotic increased (p < 0.05) the level of prophenoloxidase and g-type lysozyme gene on day 30th and 60th after treatment. On day 30th and 60th, penaeidin gene expression was significantly higher in all treatment groups compared to the control group (p < 0.05). In general, findings of this study demonstrated that oral administration of 0.5 and 1.0 g kg(-1) multi-strain probiotic improved the performance of the fish and increased the expression of immune related genes.

Protective effects of the prebiotic on the immunological indicators of rainbow trout (Oncorhynchus mykiss) infected with Aeromonas hydrophila.

The aim of this study was to investigate the protective effects of dietary administration of commercial prebiotic, Immunogen, on immunological indicators, enzymatic responses and stress tolerance in juvenile (81.65 ± 1.49) rainbow trout (Oncorhynchus mykiss) following Aeromonas hydrophila infection. The first group of fish was fed with the diet containing 2 g kg(-1) Immunogen whilst the control group received the diet free of Immunogen. There were three replicates per group. After 6 weeks feeding, the control group were divided into two treatments injected with saline buffer (control), and 1.5 × 10(8) CFU A. hydrophila respectively. The fish fed with the Immunogen supplemented diet were also injected with 1.5 × 10(8) CFU A. hydrophila. Our results revealed that dietary Immunogen increased the level of white blood cell (WBC) and percentage of lymphocyte (P < 0.05), however, the level of red blood cell (RBC), Hematocrit (Hct), hemoglobin (Hb) and percent of monocyte decreased in Untreated-Challenged group but unaffected in the group fed with Immunogen (P < 0.05). The level of lysozyme, alternative complement, antiprotease activity, total protein, albumin and globulin decreased in Untreated- Challenged group compared to control group. However, there was an increase in the level of lysozyme, alternative complement, antiprotease activity, bactericidal activity, in the Treated- Challenged group compared to other groups (P < 0.05). Serum alkali phosphatase (ALT) and aspartate aminotransferase, significantly increased fallowing challenge with A. hydrophila but in the Treated-Challenged group, there was no significant difference compared to the control group (P < 0.05). Lactate dehydrogenase (LDH) level was not different between groups (P > 0.05). Serum cortisol and glucose levels were higher in the challenge group, but these levels were lower in fish under challenge that were fed Immunogen-supplemented diet in contrast to the group fed control diet. The stress responses affected by A. hydrophila challenge (P < 0.05). Serum sodium, potassium and calcium concentration decreased by A. hydrophila exposure (P < 0.05), and Immunogen showed protection effect against this change.

Increased stocking density causes changes in expression of selected stress- and immune-related genes, humoral innate immune parameters and stress responses of rainbow trout (Oncorhynchus mykiss).

The present study investigated the effects of various stocking densities on the health status (stress and immune responses) of rainbow trout (Onchorhynchus mykiss). Juvenile rainbow trout were acclimated, placed in circular tanks under stocking densities of 10, 40 and 80 kg m(-3) and reared for 30 days. The relative expression of genes involved in stress and immunity such as HSP70, LyzII, TNF-1α, IL-1ß, IL-8 and IFN-γ1 in the head kidney was determined. Serum cortisol, ACTH, total antioxidant capacity, osmolality and lactate were measured after 30 days of culture at different stocking densities (D1:10 kg m(-3), D2: 40 kg m(-3) and D3: 80 kg m(-3)) as indices of stress responses. In addition, the effects of stocking densities on serum complement, bactericidal activity, agglutinating antibody titers, serum IgM, anti-protease activity, serum total protein and alkaline phosphatase of the fish were measured. HSP70 gene expression was significantly density-dependent upregulated in D2 and D3 densities compared to D1 (P < 0.05). Also, there was significant downregulation in expression of LyzII, TNF-1α, IL-1ß, IL-8 and IFN-γ1 in fish reared at density of either D2 or D3 (P < 0.05). In terms of stress responses, serum ACTH, cortisol and lactate level showed significant density-dependent increase (P < 0.05) while serum osmolality and total antioxidant capacity showed significant decline (P < 0.05) in fish reared at higher densities (D2 and D3) compared to fish reared at lower density (D1) (P < 0.05). Concordant with the expression of the immune-related genes, the serum complement and bactericidal activity as well as specific antibody titer against Aeromonas hydrophila, IgM and anti-protease activity decreased along with elevation of stocking density from D1 to D3 (P < 0.05). However, different stocking densities had no significant effect on serum total protein level and alkaline phosphatase activity. These results suggested that elevation of stocking densities and crowding resulted in the increase in HSP70 gene expression and the levels of selected stress responses in the serum. However, there was down-regulation of immune genes expression and decreased innate immune responses in the fish. The mRNA expression of the genes and immune parameters that were measured in this study could be helpful in monitoring the health status and welfare of the fish in aquaculture systems particularly in relation to increased stocking densities.

Dietary fermentable fiber upregulated immune related genes expression, increased innate immune response and resistance of rainbow trout (Oncorhynchus mykiss) against Aeromonas hydrophila.

This trial was carried out to investigate the effects of dietary administration of Vitacel(®), a commercial fermentable fiber, on immune related genes (Lysozyme, TNFα and HSP70) expression, innate immune response and resistance of rainbow trout against Aeromonas hydrophila. 120 healthy rainbow trout (81.65 ± 1.49 g) were distributed in six fiberglass tanks assigned to two treatments. The treatments were feeding rainbow trout with diets supplemented with 0 (control) or 10 g kg(-1) Vitacel(®) for 45 days. The results revealed that administration of fermentable fiber significantly (P < 0.05) upregulated lysozyme and TNFα gene expression. HSP70 gene expression was significantly lower in Vitacel(®) fed fish at the end of trial (P < 0.05). Furthermore dietary administrations of Vitacel(®) remarkably elevated rainbow trout innate immune parameters include serum lysozyme, ACH50, bactericidal activity and agglutination antibody titer (P < 0.05). Administration of 10 g kg(-1) Vitacel(®) significantly increased rainbow trout resistance against A. hydrophila (P < 0.05). The results of present study revealed that dietary Vitacel(®) can upregulates immune related genes expression and elevates innate immune response and disease resistance of rainbow trout.