The Optimum Dietary Phenylalanine Requirement of Hybrid Grouper (Epinephelusfuscoguttatus ♀ × Epinepheluslanceolatus ♂) Juveniles: Effects on Growth Performance, Gut Micromorphology, and Antioxidation.

The optimum phenylalanine (Phe) requirement for hybrid grouper (Epinephelusfuscoguttatus ♀ × Epinepheluslanceolatus ♂) juveniles was determined through an 8-week growth trial. A total of seven isoenergetic (340 kcal per 100 g of dry matter), isonitrogenous, and isolipidic diets were made, containing 8.2 (Phe 8.2), 9.2 (Phe 9.2), 10.1 (Phe 10.1), 11.2 (Phe 11.2), 13.3 (Phe 13.3), 15.2 (Phe 15.2), and 17.3 g/kg (Phe 17.3), respectively. Triplicate tanks of juvenile fish (about 16.7 g/fish) were fed each experimental diet twice daily until apparent satiation. The results indicated that different dietary Phe levels significantly influenced weight gain percentage (WG), feed efficiency (FE), protein efficiency ratio (PER), as well as, productive protein value (PPV). Fish fed Phe 8.2 had the lowest WG or PPV among all experimental treatments. Furthermore, the optimal dietary Phe level increased fold height, width, enterocyte, and microvillus height of fish. The Phe 10.1 group exhibited higher growth hormone (GH) expression in the pituitary compared to other groups. Expression of hepatic insulin-like growth factor-1 (IGF-1) and growth hormone receptor 1 (GHR1) displayed a similar pattern of variation to that of GH. The Phe 13.3 group had lower expression of S6 kinase 1 (S6K1) and target of rapamycin (TOR) than other groups. In addition, fish fed Phe 10.1 had lower levels of nuclear factor erythroid 2 (Nrf2) and heat shock protein 70 (HSP70) in the head kidney, and Cu/Zn-superoxide (Cu/ZnSOD) dismutases in the midgut compared to fish fed other Phe levels. Generally, optimal Phe content in the diet of hybrid grouper was estimated to be 12.7 g/kg of dry matter (27.3 g/kg of dietary protein), and at this level, the feed utilization, gut micromorphology, and immunity of fish were also elevated.

Effects of Dietary Protein-to-Energy Ratios on Growth, Immune Response, Antioxidative Capacity, Liver and Intestinal Histology, and Growth-Related Gene Expression in Hybrid Yellow Catfish (Pelteobagrus fulvidraco ♀ × Pelteobagrus vachelli ♂).

This study is aimed at evaluating the effects of dietary protein-to-energy ratios on the growth, immunological response, antioxidative capacity, liver and intestinal histology, and growth-related gene expression of hybrid yellow catfish (Pelteobagrus fulvidraco ♀ × Pelteobagrus vachelli ♂). Eight diets were formulated to form different protein/energy ratios of 84, 88, 90, 93, 95, 96, 99, and 103 mg/kcal (P/E84, P/E88, P/E90, P/E93, P/E95, P/E96, P/E99, and P/E103), respectively. These diets contain different levels of gross energy (GE), ranging from 4.13 to 4.76 kcal g-1. Seven hundred and twenty healthy fish (17.15 ± 0.02 g) were randomly dispersed into 24 rectangular fiberglass tanks with 8 treatments in triplicate groups. The fish fed a P/E ratio of 95 mg/kcal demonstrated the best growth and feed utilization. A significant (P < 0.05) increase in percent weight gain (WG%) and specific growth rate (SGR) was seen as the dietary P/E ratio ameliorated from P/E84 to P/E95, followed by a decreased pattern in these parameters. Feed conversion ratio (FCR) and daily feed intake (DFI) were significantly impacted by dietary P/E ratios (P < 0.05). Additionally, an optimum P/E ratio improved intestinal morphology. However, low or high P/E ratio diets can cause oxidative stress, impaired liver function, and significantly reduced nonspecific immunity. The expression of target of rapamycin (TOR) and insulin-like growth factor-1 (IGF1) genes in the liver was considerably influenced by dietary protein-to-energy ratios (P < 0.05). Based on the statistical analysis of WG% against the dietary P/E ratio, the optimal P/E ratio for the studied species was estimated to be 92.92 mg/kcal.

Authentication of the therapeutic Lamiaceae taxa by using pollen traits observed under scanning electron microscopy.

Herbal medicines are gaining popularity worldwide for human healthcare because of their therapeutic potential. However, adulteration and use of unauthentic raw herbals as substitutes have become a major issue for the local communities and industry for reasons of safety and efficacy. Therefore, the authentication of medicinal plants before their use in herbal medicines is a need of time. Hence, the present study was designed with an aim, to authenticate the therapeutic Lamiaceous taxa by using pollen traits observed under scanning electron microscopy. Pollen micro-morphological studies solve the problem through discrimination and correct identification of the Lamiaceae species from the adulterants. Based on pollen features, Lamiaceae were further divided into two sub-families Lamioideae (tricolpate) and Nepetoideae (hexa-colpate). The pollen grains of Lamioideae were found as small to medium-sized, tricolpate, radially/bilateral symmetrical, sub-spheroidal and oblate shape. Besides, exine patterns bireticulate, reticulate and micro-reticulate, colpus surface sculpturing as psilate, gemmate, scabrate, and verrucate have also been reported. A significant variation was found in the pattern of the reticulum, thickness, and the number of secondary lumina per primary lumen. Similarly, Nepetoideae has a hexa zono-colpate pollen but tri and tetra zono-colpate pollen have also been observed. Hence, this study contributes to the authentication and correct identification of medicinally important Lamiaceae taxa by using scanning electron microscopic techniques and can help to solve the adulteration problem. Highlights Authentication of medicinally important Lamiceous taxa was carried out through scanning electron microscopic techniques. Chemotaxonomic characterization was used for the accurate identification of the therapeutic taxa. The medicinal, palynological and phytochemical significance of Lamiaceae taxa were evaluated. A significant variation was seen in the palynological traits that help in the determination and authentication of the therapeutic Lamiaceous species. Based on the chemotaxonomic characterization, our study can help to solve the adulteration problem for the reason of safety and efficacy.

Pollen morphology and its implication in the taxonomy of some selected taxa of the bi and tri-ovulate Euphorbiaceae of the Hainan Island by using multiple microscopic techniques.

The family Euphorbiaceae is one of the most taxonomically complex and diverse family among the angiosperm. Therefore, this study aims to document the pollen morphology of the Euphorbiaceae taxa collected from different geographical regions of the Hainan Island South China, to strengthen the taxonomy and support the subfamilial classification. The Euphorbiaceae includes 23 species and 10 genera of the bi-ovulate subfamily Phyllanthoideae and tri-ovulate Crotonoideae, Euphorbioideae, and Acalyphoideae were examined under light and scanning electron microscopy. The principal component analysis was performed by using six variables to show variation and relationships between the species. A significant variation was found in the pollen morphology (exine and colpus sculpturing). The croton-type pattern of the pollen was found the diagnostic trait of the Crotonoideae. The Euhphorboideae taxa were mostly homogenous however, variation was found in the muri pattern. The Acalyphoideae, have fossulate scabrate and verrucate exine sculpturing that can delimit from the rest of sub-families. The Phyllanthoideae, showed reticulate exine, tri to tetra-colporate having various colpus morphology. The maximum pollen size was found in Croton hirtus (59.49 × 60.86 µm) and minimum in Acalypha indica (10.86 × 73 µm). Similarly, the maximum colpus width was observed in Euphorbia pulcherrima (8.3 µm). Hence pollen traits have relevance in the taxonomy and phylogenetic of the family to define species boundaries at various taxonomic levels. This study can support and strengthen the taxonomy of subfamilies and fill the palynological gap of these taxa in Hainan Island and will contribute to future systematic studies of the Euphorbiaceae.

Palynomorphological diversity among the Asteraceous honeybee flora: An aid to the correct taxonomic identification using multiple microscopic techniques.

This study aimed to characterize the palynological morphology of melliferous species of family Asteraceae belonged to seven tribes which were categorized into 15 genera and were studied under light and scanning electron microscopy. The pollen grains were acetolyzed, measured, and described qualitatively. Quantitative data were analyzed by descriptive and multivariate statistical analysis. The species were analyzed considering 11 quantitative pollen characteristics. The study revealed that the pollen grains are circular, triangular, angular, semi-angular, quadrangular, pentagonal and hexagonal amb, and 3-colporate or 3-colpate. The size of the pollen is variable among the species, ranged from 56.1 to 23.2 µm. Three pollen types: oblate spheroidal (eight species), prolate spheroidal (six species), and suboblate (two species) were observed. The surface pattern of the exine varies from echinoperforate, echinate, echinate microperforate, fenestrate, echinoperforate reticulate, microreticulate, echinate perforate, tectate, and scabrate. Exine thickness was calculated minimum in Cichorium intybus (2.47 µm) and maximum in Taraxacum campylodes (5.15 µm). A key to studied bee floral species, based on the morphological features of pollen grains, is also provided. The palynomorphological characters here analyzed can be potentially used to correctly identify the Asteraceous honeybee floral species most commonly plants visited by honeybees in the study area.

Toxicological effects of the chemical and green ZnO NPs on Cyprinus carpio L. observed under light and scanning electron microscopy.

Nanoparticles in aquatic bodies cause serious harm to the aquatic organisms when accumulated in high amounts. However, green nanoparticles synthesized using plants can be less toxic as compared to chemical nanoparticles. Hence, we designed our study to investigate the toxicological effects of chemical and green zinc oxide nanoparticles (ZnO NPs) on the biological activity of juvenile Cyprinus carpio. The green ZnO NPs were synthesized from Solieria robusta, and chemical ZnO NPs were synthesized using zinc chloride solution and ammonium hydroxide. Characterization was done by using light microscopy, scanning electron microscope (SEM), Fourier transmission infrared radiation, and X-ray diffraction (XRD) techniques. The highest absorbance of nanoparticles was observed at 360 which confirmed the synthesis of ZnO. The SEM analysis showed that green nanoparticles were hexagonal while the chemical nanoparticles were spherical to cubic in shape. Definite peaks were observed in XRD of green and chemical NPs at 2θ angles 45.84° and 32.18°, respectively. Oxidative stress was determined by chemical analysis of catalase, glutathione S-transferase (GST), glutathione (GSH), and lipid peroxidation (LPO) activities. The toxicological effects of chemical ZnO NPs on the catalase, LPO, GST, and GSH activities were more than green ZnO NPs. The histopathological investigation proved that the effect of chemical nanoparticles was worse than green ZnO NPs. More tissue damage was found in chemical nanoparticles than green synthesized nanoparticles. It was concluded that chemical nanoparticles can be replaced by green nanoparticles, as green nanoparticles are eco-friendly with less toxicological effects. This replacement can limit the toxic effect of nanoparticles when they get accumulated in high amounts in water bodies.

Chemo-taxonomic and biological potential of highly therapeutic plant Pedicularis groenlandica Retz. using multiple microscopic techniques.

Pedicularis groenlandica is one of the most important medicinal plant of Deosai Plateau (Gilgit-Baltistan) Pakistan. The present study was aim to evaluate the micromorphological features, phytochemical screening and pharmacological potential of P. groenlandica by using multiple microscopic techniques. Six different solvents were used to prepare P. groenlandica extracts. Phytochemical and antioxidant activities were determined calorimetrically. To investigate antidiabetic, α-amylase inhibition assay was performed. Cytotoxicity was tested using brine shrimp assay. Anti-leishmanial via MTT assay. Disc-diffusion assay was used for protein kinase inhibitory, antibacterial and antifungal activities. Palyno-anatomical study showed significant variation for the authentication and correct identification of this highly therapeutic plant by using light and scanning electron microscopic techniques. All extracts were found rich in phytochemicals, significant amount of phenolic and flavonoid contents were found in methanol extract (PGM) 95.78 mg GAE/g and 66.90 mg QE/g. Highest DPPH scavenging potential with IC50 88.65 µg/mL, total antioxidant capacity (60.33 mg AAE/g sample) and total reducing power (83.97 mg AAE/g) were found for PGM. Disc-diffusion method showed significant antibacterial and antifungal activities. Noticeable growth inhibition in L. tropica was displayed by n-hexane extract (IC50 112 µg/mL). Brine shrimp with highest LD50 (67.65 µg/mL) in ethyl-acetate extract. Ethanol extract gives persuasive protein kinase inhibition (26 mm) against Streptomyces 85-E hyphae. Highest alpha-amylase inhibition (74.10%) was found in n-hexane extract. In conclusion, our findings scientifically support the ethno-medicinal and biological potential of P. groenlandica. In future, the plant needs to be explored for further identification and isolation of bioactive compounds to develop new drugs to treat several aliments.

Effects of Dietary Carbohydrate to Lipid Ratios on Growth Performance, Muscle Fatty Acid Composition, and Intermediary Metabolism in Juvenile Black Seabream (Acanthopagrus schlegelii).

An 8-week feeding trial was conducted to evaluate the effects of dietary carbohydrate to lipid (CHO:L) ratios on growth performance, muscle fatty acid composition, and intermediary metabolism in juvenile black seabream (Acanthopagrus schlegelii). Five isonitrogenous and isoenergetic diets (48.0% crude protein and 18.0 MJ kg-1 gross energy) were formulated to contain different CHO:L ratios ranging from 0.33 to 3.75. Triplicate groups of 20 fish averaging 0.51 ± 0.01 g were fed with experimental diets twice daily to apparent satiation. The results indicated that final body weight (FBW), percentage weight gain (PWG), specific growth rate (SGR), and protein efficiency ratio (PER) were significantly influenced by the dietary CHO:L ratios (p < 0.05). The highest FBW, PWG, and SGR were observed in fish fed the diet with a CHO:L ratio of 1.36 (p < 0.05). A two-slope broken-line regression analysis based on PWG indicated that the optimal dietary CHO:L is 1.08. Lipid content in the whole body decreased, and glycogen concentration in the liver increased with the increase of dietary CHO:L ratios from 0.33 to 3.75 (p < 0.05). Moreover, there was a positive correlation between muscle fatty acid composition and dietary fatty acid composition. The relative expression levels of genes involved in glucose metabolism, such as gk, pepck, and glut2 were upregulated by increasing the dietary CHO:L ratio. Also, the mRNA expression level of genes related to lipid synthesis, such as fas and accα were significantly upregulated with dietary CHO:L ratios increasing from 0.33 to 3.75. The highest expression of genes involved in fatty acid ß-oxidation, such as cpt1 and acox1, were observed in fish fed the 1.36 CHO:L ratio diet. The gene expression of Δ6 fatty acyl desaturase (fads2) in the liver significantly increased with increase of dietary CHO:L ratios from 0.33 to 3.75. Fish fed the diet with CHO:L ratios of 2.26 and 3.75 had lower expression levels of elovl5 than those fed the other diets. These results demonstrate that dietary optimal CHO:L ratios could improve PWG and SGR but also influence expression of genes involved in glucose and lipid metabolism. Based on the overall results, the optimal dietary CHO:L ratio is 1.08 for black seabream.

Influence of dietary replacement of fish meal with fish soluble meal on growth and TOR signaling pathway in juvenile black sea bream (Acanthopagrus schlegelii).

An 8-week feeding trial was conducted to evaluate the effect of replacement of fish meal (FM) with fish soluble meal (FSM) on growth performance, feed utilization and expression of genes involved in TOR signaling pathway for juvenile black sea bream (Acanthopagrus schlegelii). Six isonitrogenous (41%) and isolipidic diets were prepared to contain graded levels of FSM which replaced 0% (control diet), 10%, 20%, 30%, 40% and 60% protein from FM. Triplicate groups of 20 fish with initial weight 0.51 ± 0.01 g were fed with experimental diets twice daily to apparent satiation. The results showed significant differences in growth performance and feed utilization among all treatments, final body weight (FBW), percent weight gain (PWG), specific growth rate (SGR) and protein efficiency ratio (PER) significantly increased with dietary replacement levels of FM with FSM increasing from 0% to 40% (P < 0.05), PWG, SGR and PER were significantly reduced when replacement of FM with FSM further increased from 40% to 60%. Based on PWG against replacement levels of FM with FSM, A two-slope broken-line model analysis indicated that the optimal replacement of FM with FSM is to be 42.59%. Moreover, the lowest feed conversion ratio (FCR) was observed in fish fed the 40% FSM replacement diet. Muscle amino acid profile in muscle revealed that total essential amino acids, arginine and threonine were significantly influenced by replacement levels of FSM, while there was no significant difference in NEAA among all treatments. The hematological indices were not affected by the replacement levels of FM with FSM. The relative expression levels of irs-1, pi3k, akt, igf-1, s6k1 and tor were up-regulated when replacement levels of FM with FSM increased from 0% to 40%, and higher values were observed in fish fed with 40% FSM replacement diet compared to those fed the other diets. However, relative expression of 4e-bp2 was down-regulated when replacement levels of FM with FSM increased from 0% to 40% (P < 0.05). In summary, the results of present study indicated that FSM could be a viable alternative protein source for black sea bream, dietary FSM supplementation could improve growth and up-regulate the relative expression of irs-1, pi3k, akt, igf-1, s6k1 genes related to TOR signaling pathway in liver of juvenile black sea bream.

The effects of dietary yeast hydrolysate on growth, hematology, antioxidant enzyme activities and non-specific immunity of juvenile Nile tilapia, Oreochromis niloticus.

The present study was aimed to compare and evaluate the impacts of supplemented diets with different yeast hydrolysate (YH) levels on growth performance, body composition, hematological characteristics, antioxidant enzyme activities, and non-specific immunity (intestinal cytokines) of juvenile Nile tilapia (Oreochromis niloticus). Three isonitrogenous (protein, 33%) and isolipidic (lipid, 6%) experimental diets supplemented graded levels of YH (0% for control; 1% and 3% as tested diets) were fed to juvenile Nile tilapia. A total of 240 fish with initial body weight averaging 3.5 ± 0.02 g were randomly divided into three groups with four replicates per group and 20 fish for each replicate. For apparent satiation, the fish were fed twice daily during eight weeks. The results showed no significant difference in survival among all treatments. The fish fed the diet containing 1% yeast hydrolysate had significantly elevated weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER) compared to the control group and lower feed conversion ratio (FCR). The fish fed 1% and 3% YH showed higher glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT) activity and a significantly lower malondialdehyde (MDA) level in the liver than the control group, indicating enhancement of the anti-oxidant status. Serum lysozyme activity was significantly increased in the diet having 1% and 3% yeast hydrolysate supplementation groups, suggesting an improvement influence on the non-specific immune response. The expression of IL-1ß, IL-10, TNF-α, TGF-ß2, ALP and TLR2 was significantly elevated in fish fed the diet containing 1% YH. In conclusion, dietary supplementation with 1% yeast hydrolysate improves growth performance, and feed utilization enhances the antioxidant status and exerts an adequate stimulus on the non-specific immunity (intestinal cytokines) of Nile tilapia.