Effects of aqueous and ethanolic leaf extracts from drumstick tree (Moringa oleifera) on gilthead seabream (Sparus aurata L.) leucocytes, and their cytotoxic, antitumor, bactericidal and antioxidant activities.

Aqueous and ethanolic extracts of drumstick, Moringa oleifera, leaves were evaluated in vitro to ascertain their principal active components and determine their immunostimulant, cytotoxic, antitumoral, bactericidal and antioxidant activities. Phytochemical screening of M. oleifera leaf extracts showed a greater abundance of phenolic and cyanogenic glycosides in aqueous than in ethanolic extracts, characterized by several flavonoids, condensed tannins and saponins. No significant effects on gilthead seabream (Sparus aurata) head-kidney leucocyte activities (phagocytic ability and capacity, respiratory burst and peroxidase) were detected after incubation for 24 h with different concentrations (0.001/1 mg mL-1) of either extract. In addition, the aqueous extract showed a marked cytotoxic effect on both SAF-1 (at doses above 0.01 mg mL-1) and PLHC-1 (at doses above 0.25 mg mL-1) cell lines. The ethanolic extract improved the viability of SAF-1 cells and decreased the viability of PLHC-1 cells when used at higher concentrations. Both the ethanolic and, particularly, the aqueous extracts showed significant bactericidal activity on pathogenic Vibrio anguillarum and Photobacterium damselae strains. The antiradical activity of M. oleifera, as determined by the ABTS assay, increased in a linear dose-response with increasing extract concentrations. The results as a whole for the cytotoxic, bactericidal and antioxidant activities of M. oleifera leaf extracts point to their possible use as additives in functional diets for farmed fish.

Dietary supplementation of drumstick tree, Moringa oleifera, improves mucosal immune response in skin and gills of seabream, Sparus aurata, and attenuates the effect of hydrogen peroxide exposure.

In aquatic animals, the mucosal barrier is the first line of innate immune defence against external chemicals and pathogens. In this study, the effects of dietary Moringa oleifera leaf (MOL) supplementation on skin and gill mucosal immunity, antioxidants and stress responses were evaluated in seabream (Sparus aurata) fingerlings exposed to hydrogen peroxide (H2O2). A total of 144 specimens (10.11 ± 0.41 g) were divided into four treatments (three replicates per treatment contained 12 specimens each) and fed a non-supplemented control diet or a 1, 2.5 or 5% MOL-supplemented diet. After three weeks of feeding, six specimens from each aquarium were sampled for blood, mucus and tissues. The other six fish in each aquarium were subjected to H2O2 exposure. The results revealed that MOL did not negatively affect either cortisol or glucose levels. MOL supplementation significantly (P < 0.05) improved skin mucosal immunity-related characteristics, including phosphatase, peroxidase and lysozyme activity and IgM levels. Additionally, MOL upregulated the expression of antioxidant genes (sod and cat), an anti-inflammatory gene (tgf-ß), tight junction protein genes (occludin and zo-1), c3, and igm in both the skin and gills. However, H2O2 exposure significantly (P < 0.05) increased both cortisol and glucose levels and disrupted skin mucosal immune function by significantly (P < 0.05) decreasing phosphatase, peroxidase, protease, antiprotease and lysozyme activity and IgM levels. H2O2 exposure severely decreased the mRNA levels of the studied genes. MOL dietary supplementation at the 5% level successfully attenuated the negative effects of H2O2 on the mucosal immune response in both the skin and gills. In conclusion, dietary MOL supplementation at the 5% level is recommended to improve S. aurata mucosal immune function under both normal and stress conditions. Additionally, exposure to H2O2 disrupts the mucosal immunity of fish. This contributes knowledge on the routes involved in mucosal innate immunity and could help to understand the fish resistance against chemicals exposure. Graphical abstract.

The Optimization of Dietary Protein Level and Carbon Sources on Biofloc Nutritive Values, Bacterial Abundance, and Growth Performances of Whiteleg Shrimp (Litopenaeusvannamei) Juveniles.

A biofloc technology-based 75-day indoor growth trial in an 80 L glass aquaria was conducted to evaluate the effects of two different carbon sources (sugarcane bagasse, SB, and wheat flour, WF) on the biofloc composition, bacterial abundance, and growth of whiteleg shrimp (Litopenaeus vannamei) juveniles (0.23 ± 0.04 g). Three different levels of dietary protein content (250, 300, and 350 g protein kg−1 diet) and two carbon sources (SB and WF) were applied (SB250, WF250, SB300, WF300, SB350, and WF350, respectively), comparing to a controlled diet without biofloc and fed on a 450 g protein kg−1 diet (C450). With the addition of SB and WF, water quality was in the ideal recommended ranges for L. vannamei culture. At the end of the experiment, the biofloc volume increased with increasing dietary protein levels. The nutritional value of biofloc in different treatments was influenced by dietary protein and added SB and WF. Increasing dietary protein significantly increased the protein and lipid contents of the produced biofloc. The use of WF as a carbon source significantly increased lipids and nitrogen-free extract in the biofloc. The total heterotrophic bacterial (THB) count was significantly higher (p < 0.05) in WF300 and WF350 than in the other treatments. The mean effect of the protein levels and carbon source was significantly reported, whereas the highest significant THB count was recorded with 300 dietary protein and using WF as a carbon source. The growth performances of L. vannamei fed with biofloc treatments were significantly (p < 0.05) higher than the C450 group. The highest final weight and weight gain were recorded in SB350 treatment. The feed conversion ratio was not affected by reducing dietary protein levels; meanwhile, the protein efficiency ratio increased significantly in biofloc treatments than in the control. Overall, the results demonstrate that, compared to the control treatment of 450 dietary protein, the biofloc treatments using WF as a carbon source could compensate for the reduction in the dietary protein levels in the diet of L. vannamei and maintain higher zootechnical performance.

Growth Performance, Immune-Related and Antioxidant Genes Expression, and Gut Bacterial Abundance of Pacific White Leg Shrimp, Litopenaeus vannamei, Dietary Supplemented With Natural Astaxanthin.

The current study examines the effect of dietary supplementation of ethanolic extract of Arthrospira platensis NIOF17/003, which is mainly natural astaxanthins (97.50%), on the growth performance, feed utilization, bacterial abundance, and immune-related and antioxidant gene expressions of the Pacific white leg shrimp, Litopenaeus vannamei. A total of 360 healthy L. vannamei postlarvae (0.19 ± 0.003 g) were divided into four groups (0, 2, 4, and 6 g natural astaxanthins/kg diet) each in three replicates, at an initial density of 30 PLs per tank (40 L capacity). The shrimp were fed the tested diets three times a day at a rate of 10% of their total body weight for 90 days. Diets supplemented with different astaxanthin levels significantly improved shrimp growth performance and feed conversion ratio compared to the control diet. No significant differences were observed in survival rates among all experimental groups. The immune-related genes (prophenoloxidase, lysozyme, beta-glucan binding protein, transglutaminase, and crustin) mRNA levels were significantly upregulated in groups fed with different concentrations of the natural astaxanthins in a dose-dependent manner. The prophenoloxidase gene is the highest immune-upregulated gene (14.71-fold change) in response to astaxanthin supplementation. The superoxide dismutase mRNA level was significantly increased with increasing dietary astaxanthin supplementation. In addition, increasing astaxanthin supplementation levels significantly reduced the count of heterotrophic bacteria and Vibrio spp. in the culture water and shrimp intestine. Overall, the current results concluded that diet supplementation with natural astaxanthin, extracted from Arthrospira platensis, enhanced the growth performance, immune response, and antioxidant status of L. vannamei.

Ginseng, Tribulus Extracts and Pollen Grains Supplementation Improves Sexual State, Testes Redox Status, and Testicular Histology in Nile Tilapia Males.

This study aimed to investigate the effect of dietary supplementation of three natural antioxidants on sex hormone levels, enzymatic and non-enzymatic antioxidant systems, and histological changes in the testes of male Nile tilapia, Oreochromis niloticus. A total of 210 male Nile tilapia were distributed into seven treatments (three replicates for each) with an initial weight of 3.67 g fish-1. The fish were fed experimental diets (32% crude protein) without supplementation as control or supplemented with ginseng extract (GE; 0.2 and 0.4 g GE kg-1 diet), Tribulus terrestris extract (TT; 0.6 and 1.2 g TT kg-1 diet), and date palm pollen grains (DPPG; 3 and 6 g DPPG kg-1 diet) for 84 days. The results revealed a significant increase in the luteinizing hormone level with TT, DPPG, and GE supplementation increased the levels by 22.9%, 18.5%, and 17.6%, respectively. The testosterone level also increased significantly with TT1.2, GE0.4, TT0.6, and DPPG6 by 86.23%, 64.49%, 57.40%, and 24.62%, respectively. The antioxidant status in the testis homogenate showed a significant decrease in the level of thiobarbituric acid-reactive substances when using different dietary substances. In addition, glutathione reduced contents, glutathione S-transferases, glutathione peroxidase, catalase, and superoxide dismutase activities significantly increased with different dietary supplementation in a dose-dependent manner. The histological evaluation revealed normal histological features of the testes in all treatments with increasing active seminiferous tubules (%) in GE, TT, and DPPG supplemented groups, especially with the highest levels. In conclusion, the dietary supplementation of GE, TT, and DPPG enhanced sex hormones level, redox status, and testis structure and could improve the male reproductive performance of Nile tilapia.

Salt-Tolerant Phenomena, Sequencing and Characterization of a Glyoxalase I (Jojo-Gly I) Gene from Jojoba in Comparison with Other Glyoxalase I Genes.

Plant response to salt stress and the mechanism of salt tolerance have received major focus by plant biology researchers. Biotic stresses cause extensive losses in agricultural production globally, but abiotic stress causes significant increase in the methylglyoxal (MG) level of GlyoxalaseI (Gly I). Identification of salt-tolerant genes when characterizing their phenotypes will help to identify novel genes using polymerase chain reaction (PCR) to amplify the DNA coding region for glyoxalase I. This method is specific, requiring only genomic DNA and two pairs of PCR primers, and involving two successive PCR reactions. This method was used rapidly and easily identified glyoxalase I sequences as salt-tolerant genes from Jojoba (Simmondsia chinensis (Link) Schneider). In the present study, the glyoxalase I gene was isolated, amplified by PCR using gene-specific primers and sequenced from the jojoba plant, then compared with other glyoxalase I sequences in other plants and glyoxalase I genes like in Brassica napus, ID: KT720495.1; Brassica juncea ID: Y13239.1, Arachis hypogaea; ID: DQ989209.2; and Arabidopsis thaliana L, ID: AAL84986. The structural gene of glyoxalase I, when sequenced and analyzed, revealed that the uninterrupted open reading frame (ORF) of jojoba Gly I (Jojo-Gly I) spans 775 bp, corresponding to 185 amino acid residues, and shares 45.2% amino acid sequence identity to jojoba (Jojo-Gly I). The cloned ORF, in a multicopy constitutive expression plasmid, complemented the Jojo-Gly I, confirming that the encoded Jojo-Gly I in jojoba showed some homology with other known glyoxalase I sequences of plants.