Exploring the impact of nano-Se and nano-clay feed supplements on interleukin genes, immunity and growth rate in European Sea Bass (Dicentrarchus labrax).

This study aimed to investigate the effects of adding Nano-Selenium (NSe) and Nano-clay (NC) as feed supplements on European Sea Bass (Dicentrarchus labrax). Two separate experiments were conducted, one with NC and the other with NSe. Each experiment consisted of four sub-groups with varying concentrations of NC or NSe. The expression levels of five immune-related genes (TNF-α, TNF-ß, IL-2, IL-6 and IL-12) were measured using Real-time Quantitative PCR (Rt-PCR) Assay. The results showed an increase in the expression of interleukins (IL-2, IL-6 and IL-12) and pro-inflammatory cytokines (TNF-α and TNF-ß) after exposure to NC and NSe. TNF-α gene expression was significantly higher with both 1 mg and 10 mg concentrations of NC and NSe. TNF-ß gene expression was highest with the 5 mg concentration of NC. The concentrations of 1 mg and 10 mg for NC, and 1 mg, 5 mg, and 10 mg for NSe, led to the highest (p < 0.05) levels of IL-2 expression compared to the control. Similar trends were observed for IL-6 and IL-12 gene expression. Understanding the impact of these concentrations on gene expression, growth rate, biochemical indices, and antioxidant status can provide valuable insights into the potential applications of NC and NSe supplements on European Sea Bass.

Toxicity Evaluation, Oxidative, and Immune Responses of Mercury on Nile Tilapia: Modulatory Role of Dietary Nannochloropsis oculata.

The current study evaluated the potential ameliorative effect of a dietary immune modulator, Nannochloropsis oculata microalga, on the mercuric chloride (HgCl2)-induced toxicity of Nile tilapia. Nile tilapia (45-50 g) were fed a control diet or exposed to » LC50 of HgCl2 (0.3 mg/L) and fed on a medicated feed supplemented with N. oculata (5% and 10% (50 or 100 g/kg dry feed)) for 21 days. Growth and somatic indices, Hg2+ bioaccumulation in muscles, and serum acetylcholinesterase (AChE) activity were investigated. Antioxidant and stress-related gene expression analyses were carried out in gills and intestines. Histopathological examinations of gills and intestines were performed to monitor the traits associated with Hg2+ toxicity or refer to detoxification. Hg2+ toxicity led to significant musculature bioaccumulation, inhibited AChE activity, downregulated genes related to antioxidants and stress, and elicited histopathological changes in the gills and intestine. Supplementation with N. oculata at 10% was able to upregulate the anti-oxidative-related genes while downregulated the stress apoptotic genes in gills and intestines compared to the unexposed group. In addition, minor to no histopathological traits were detected in the gills and intestines of the N. oculata-supplemented diets. Our data showed the benefit of dietary N. oculata in suppressing Hg2+ toxicity, which might support its efficacy as therapeutic/preventive agent to overcome environmental heavy metal pollution in aquatic habitats.

Effects of L-ascorbic acid on growth, non-specific immunity, antioxidant capacity, and intestinal and hepatopancreatic histology of red swamp crayfish, Procambarus clarkii.

This study investigated the dietary effects of coated L-ascorbic acid (LA) on growth, feed utilization, survival, serum biochemical indices, immunity, antioxidant capacity, and intestinal and hepatopancreatic histology of the pre-adult red swamp crayfish. Four isoproteinous and isolipidic diets were formulated to contain several LA levels as 0, 1300, 1600, and 1900 mg/kg and designated as control (LA0), LA13, LA16, and LA19, respectively. However, the analyzed LA concentrations in diets were 0.00, 199.57, 360.45, and 487.50 mg/kg in LA0, LA13, LA16, and LA19, respectively. Triplicate treatments of crayfish (21.60 ± 0.14 g) were fed the test diets and reared in fiberglass tanks with a density of 20 individuals per each for eight weeks. Results revealed that all LA treatments had significantly enhanced growth performance compared to the control. Of interest, the LA16 treatment recorded the highest final tank biomass, biomass gain, total feed intake, condition factor, and muscle yield among the other treatments. The tank feed conversion ratio was significantly decreased in LA treatments compared to the control. Moreover, dietary LA16 and LA19 had significantly higher survival rates (93.3%) compared to (85.0%) in the LA0 group. All dietary doses of LA significantly increased serum parameters (total protein, albumin, globulin, lysozyme activity) and respiratory burst activity compared to the LA0 treatment. Dietary LA16 significantly boosted the hepatopancreatic antioxidant capacity, manifested by decreased malondialdehyde concentrations, increased catalase, superoxide dismutase, and glutathione peroxidase enzyme activities, and reduced glutathione content compared to the LA-free diet. A normal histoarchitecture of the hepatopancreatic tubules was found in all LA treatments except with some minor degenerative changes in the tubular lumen, and hepatopancreatic cells associated with enlarged nuclei were found in the LA19. However, normal intestinal histoarchitecture was found in all treatments with no recorded intestinal lesions. Of interest, the polynomial regression performed on the analyzed LA concentrations suggested that 380 mg/kg would be suitable to provide maximal biomass gain for pre-adult crayfish. In conclusion, results revealed that coated LA could enhance the growth, immunity, and antioxidant capacity of pre-adult red swamp crayfish, suggesting its potential as a functional and necessary micronutrient for crayfish diets.

Effects of exogenous bile acids (BAs) on growth, lipid profile, digestive enzymes, and immune responses of thinlip mullet, Liza ramada.

An eight-week trial was designed to explore the dietary effects of commercially purchased exogenous bile acids (BAs) on growth, whole-body composition, lipid profile, intestinal digestive enzymes, liver function enzymes, oxidative stress biomarkers, and serum immunity of thinlip mullet, Liza ramada. Four triplicate groups (10.50 ± 0.05 g) were fed four soybean meal (SBM)-based diets supplied with several BAs levels at 0 (control), 50, 130, or 350 mg/kg feed. Results indicated that the growth was significantly increased in groups fed BAs-based diets, especially at 130 mg/kg feed. The body composition analysis showed that feeding fish on diets supplied with BAs up to 130 mg/kg decreased moisture (%) alongside increased crude protein (%). However, the body composition of fish fed a diet with 350 mg BAs/kg had the lowest moisture (%) and the highest crude protein (%). Moreover, there were significant increases in the intestinal (protease, α-amylase, and lipase) enzyme activities in the groups supplied with BAs up to 130 mg BAs/kg. Liver function enzymes (aspartate aminotransferase and alanine aminotransferase enzyme activities) were significantly decreased in BAs-supplemented groups compared to those fed the BAs-free group. On the other hand, the control group had higher total cholesterol, triglycerides, and low-density lipoprotein alongside the lower high-density lipoprotein than BAs-supplemented groups, especially at 350 mg BAs/kg feed. BAs significantly decreased hepatic malondialdehyde concentrations and increased the activity of hepatic catalase, superoxide dismutase, and total antioxidant capacity compared with those reared on the control diet. Serum lysozyme, respiratory burst, and alternative complement activities were significantly increased in BAs-supplied groups, particularly in the group supplied with 130 mg BAs/kg compared to those fed on the control diet. Accordingly, our findings recommend that including 130 mg BAs/kg in an SBM-based diet enhanced the growth, digestive enzyme activities, and liver functions, alleviated oxidative stress, boosted serum immunity, and lowered lipid metabolites in thinlip mullet. These findings will be beneficial for improving the quality of feed prepared for feeding mullets and an effective alternative strategy to support mullet farming.

The role of antifreeze genes in the tolerance of cold stress in the Nile tilapia (Oreochromis niloticus).

BACKGROUND: Tilapia is one of the most essential farmed fishes in the world. It is a tropical and subtropical freshwater fish well adapted to warm water but sensitive to cold weather. Extreme cold weather could cause severe stress and mass mortalities in tilapia. The present study was carried out to investigate the effects of cold stress on the up-regulation of antifreeze protein (AFP) genes in Nile tilapia (Oreochromis niloticus). Two treatment groups of fish were investigated (5 replicates of 15 fish for each group in fibreglass tanks/70 L each): 1) a control group; the fish were acclimated to lab conditions for two weeks and the water temperature was maintained at 25 °C during the whole experimental period with feeding on a commercial diet (30% crude protein). 2) Cold stress group; the same conditions as the control group except for the temperature. Initially, the temperature was decreased by one degree every 12 h. The fish started showing death symptoms when the water temperature reached 6-8 °C. In this stage the tissue (muscle) samples were taken from both groups. The immune response of fish exposed to cold stress was detected and characterized using Differential Display-PCR (DD-PCR). RESULTS: The results indicated that nine different up-regulation genes were detected in the cold-stressed fish compared to the control group. These genes are Integrin-alpha-2 (ITGA-2), Gap junction gamma-1 protein-like (GJC1), WD repeat-containing protein 59 isoform X2 (WDRP59), NUAK family SNF1-like kinase, G-protein coupled receptor-176 (GPR-176), Actin cytoskeleton-regulatory complex protein pan1-like (PAN-1), Whirlin protein (WHRN), Suppressor of tumorigenicity 7 protein isoform X2 (ST7P) and ATP-binding cassette sub-family A member 1-like isoform X2 (ABCA1). The antifreeze gene type-II amplification using a specific PCR product of 600 bp, followed by cloning and sequencing analysis revealed that the identified gene is antifreeze type-II, with similarity ranging from 70 to 95%. The in-vitro transcribed gene induced an antifreeze protein with a molecular size of 22 kDa. The antifreeze gene, ITGA-2 and the WD repeat protein belong to the lectin family (sugar-protein). CONCLUSIONS: In conclusion, under cold stress, Nile tilapia express many defence genes, an antifreeze gene consisting of one open reading frame of approximately 0.6 kbp.

Nutritional and immunological evaluation of Nannochloropsis oculata as a potential Nile tilapia-aquafeed supplement.

Nannochloropsis oculata (N. oculata) is a marine microalga containing bioactive compounds and a high omega-3 polyunsaturated fatty acid (ω-3 PUFAs). Therefore, it is very promising for nutraceutical and the functional food industry applications. Three groups of Nile tilapia (forty-five fish/group) were fed on basal diets or diets containing 5% (N5) or 10% (N10) of the microalga N. oculata for seven weeks. Fish growth performance, proximate composition, and lipid (fatty acids/ FAs and lipoproteins) profile were estimated. In addition, the expression pattern of some lipid metabolism and immune-relevant genes were assessed. An enhancement in whole body crude protein and growth indices of Nile tilapia was observed on both the supplemented groups N5 and N10. Higher levels of high-density lipoproteins (HDL); and lower levels of the low-density lipoproteins (LDL) were evident in both supplemented groups, while the cholesterol and triglycerides (TG) levels were similar among groups. Ω-3 PUFAs were the significant FAs profile of tilapia fed on N. oculata-supplemented diets in terms of eicosapentaenoic acid, docosahexaenoic acid, and n3/n6 ratio. Concerning the gene expression pattern, heat-shock protein70, glutathione-S-transferase, glutathione peroxidase, and interleukin-1ß (IL-1ß) were elevated significantly in both supplemented groups. IL-10 is only upregulated in the N10 group. The lipid metabolism-related gene expression showed downregulation of only fatty acid synthase (FAS) in both supplemented groups, with no statistical changes in Peroxisome proliferator-activated receptor alpha (PPARα). Tumor necrosis factor-α (TNF-α), Transforming growth factor-ß1 (TGF-ß1), and the apoptotic related genes [caspase3 and Proliferating cell nuclear antigen (PCNA)] showed insignificant changes among groups. The histopathological examination of the intestine, liver, and spleen supports our findings and confirms the benefits and safeness of N. oculata dietary inclusion. Collectively, N. oculata is a very promising nutraceutical for improving fish health and sustainability of aquaculture production.

DNA barcoding of marine macroalgae as bioindicators of heavy metal pollution.

Metal pollution in the marine coastal environment is an important topical issue. In this study, the water quality in five locations along the Alexandria coast (Eastern Harbor, El-Tabia pumping station, El Mex Bay, Sidi Bishir, and Abu Talat) was assessed by measuring physicochemical parameters from water samples. Depending on the morphological classification of macroalgae, the collected morphotypes were related to Ulva fasciata, Ulva compressa, Corallina officinalis, Corallina elongata, and Petrocladia capillaceae. Corallina officinalis and Corallina elongata demonstrated a high capacity for Cd, Pb, and Ni accumulation, and the highest values of Fe, Cu, and Mn were reported in Ulva fasciata and Ulva compressa. Two standard markers were applied, and results showed that the morphological classification matched the molecular data. Moreover, the analysis of algae can only reflect the accumulation of metals. The conclusion is that Ulva compressa and Corallina officinalis are potentially suitable indicators of localized short-term heavy metal pollution.

Growth performance, antioxidant activities, and immunological responses of hapa-reared thinlip mullet (Liza ramada) juveniles fed on diets supplemented with spirulina (Arthrospiraplatensis).

Spirulina (Arthrospira platensis) (SP) has been utilized for a long time as a valued feed supplement because of its proteinous content and other beneficial phytochemical compounds. Herein, we investigated the influences of SP-supplemented diets on growth, body somatic indices, digestive enzymes, hepatic antioxidant activities, and immunological responses of hapa-reared thinlip mullet (Liza ramada) juveniles. Fish were assigned in six triplicate groups and were fed for consecutive 60 days on the prepared experimental diets containing varying SP levels as 0.0, 2.0, 4.0, 6.0, 8.0, and 10.0 g/kg diet and defined as control (CNT or SP0), SP2, SP4, SP6, SP8, and SP10 groups, respectively. The results indicated that dietary SP supplementation linearly and quadratically improved the fish growth performance, and the highest growth indices were found in the SP8 group. However, dietary SP supplementation did not significantly alter feed conversion ratio (FCR), survival rate (%), hepato-somatic index, and viscera-somatic index among all experimental groups. Meanwhile, digestive enzymes (lipase, α-amylase, and proteases) in the mid-intestine were also linearly and quadratically increased in all SP-fed groups, and their uppermost values were noted in the SP8 group. Hepatic antioxidants such as superoxide dismutase, catalase, and total antioxidant capacity in SP-supplemented groups were significantly elevated than the CNT group. Conversely, hepatic malondialdehyde contents were decreased significantly along with increasing dietary SP-supplementation levels. The immunological parameters such as lysozyme, respiratory burst, and alternative complement activities were significantly elevated in SP-fed groups than in the CNT group. These findings evoked that feeding SP-supplemented diets (especially at 8.0 g/kg diet) significantly promoted the growth, digestive enzymes, hepatic antioxidant status, and immunity of L. ramada juveniles.

Toxicity, inflammatory and antioxidant genes expression, and physiological changes of green synthesis silver nanoparticles on Nile tilapia (Oreochromis niloticus) fingerlings.

The rapid increase of incorporating silver nanoparticles (Ag-NPs) in different anthropogenic and industrial activities increased the discharge of these particles in the aquatic ecosystem. The environmental impact of Ag-NPs, especially the green synthesized is still not completely understood on fish. Therefore, this study aimed to investigate the effects of exposure to graded series of starch-mediated Ag-NPs at levels of 0, 3.31, 6.63, 13.25, and 26.50 mg L-1 representing 0, 6.25, 12.5, 25, and 50% of LC50 on Nile tilapia (O. niloticus), respectively. Fish with initial weight 37.63 ± 0.41 g were maintained in 70 L glass aquaria and exposed to starch-mediated Ag-NPs (average particle size 40 nm) for 28 days. The results revealed that starch-mediated Ag-NPs induced severe changes in the mRNA levels of toxicity (CYP1A and Hsp70) and inflammatory (TNF-α and TGF-ß) genes. The expression of antioxidant genes (SOD and CAT) was significantly suppressed, and the activities of their enzymes were inhibited significantly upon exposure. Simultaneously, the malondialdehyde level increased significantly with increasing the exposure levels of starch-mediated Ag-NPs. The red blood cells, hemoglobin, hematocrit and white blood cell values were decreased significantly with doses over 3.31 mg L-1 of Ag-NPs. In addition, the total protein and globulin decreased significantly with increasing Ag-NPs in a dose-dependent manner. The liver function enzymes and kidney function indicators revealed severe toxicity with Ag-NPs exposure. In conclusion, the effect of starch-mediated Ag-NPs in doses over 3.31 mg L-1 induced obvious toxicity in the molecular and proteomic levels in Nile tilapia fingerlings.