ABSTRACT
The main objective of this study was to investigate the effects of freshly sprouted barley on the growth of lambs, in addition to its nutritional value and digestibility. In addition, sprouted barley digestibility and rumen fermentation were studied in vitro on a dry matter (DM) basis. A total of 45 three-month-old Awassi lambs were randomly assigned to five treatments of sprouted barley (0, 25, 50, 75, 100%) diets. Bodyweight, weight gain, feed intake and feed efficiency were recorded every two weeks. Nutrient analyses were performed on feed, faecal, and urine samples. DM and non-fibrous carbohydrates were measured. Digestibility of DM, organic matter (OM), and neutral detergent fiber (NDF), as well as gas production, pH value, ammonia-N, and volatile fatty acids (VFAs), were determined in vitro using continuous culture. The results showed that final bodyweight was lower (p < 0.05), while feed intake and the feed-to-gain ratio were increased (p < 0.05) in sprouted barley treatments. Nutrient analysis indicators of sprouted barley treatments (25 to100%) were lower (p < 0.05) for DM, crude protein, acid detergent fiber, lignin and ash, and higher for total digestible nutrients, NDF, fat, phosphorus, zinc, copper, and net energy than the traditional diet. In the in vivo study, the digestibility of nutrients in sprouted barley treatments was improved (p < 0.05), while the diet (sprouted barley 100%) had the lowest digestibility of DM, OM, and NDF compared with the other treatments in the in vitro study. In conclusion, the addition of sprouted barley improved digestibility, and fermentation characteristics, while having a negative effect on growth. Further studies are recommended for optimal growth performance.
ABSTRACT
Molybdenum ions (Mo) can improve plants' nutritional value primarily by enhancing nitrogenous metabolism. In this study, the comparative effects of seed priming using Mo were evaluated among sproutings of Canavalia species/cultivars, including Canavalia ensiformis var. gladiata (CA1), Canavalia ensiformis var. truncata Ricker (CA2), and Canavalia gladiata var. alba Hisauc (CA3). Mo impacts on growth, metabolism (e.g., nitrogen and phenolic metabolism, pigment and total nutrient profiles), and biological activities were assayed. Principal component analysis (PCA) was used to correlate Mo-mediated impacts. The results showed that Mo induced photosynthetic pigments that resulted in an improvement in growth and increased biomass. The N content was increased 0.3-fold in CA3 and 0.2-fold in CA1 and CA2. Enhanced nitrogen metabolism by Mo provided the precursors for amino acids, protein, and lipid biosynthesis. At the secondary metabolic level, phenolic metabolism-related precursors and enzyme activities were also differentially increased in Canavalia species/cultivars. The observed increase in metabolism resulted in the enhancement of the antioxidant (2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical scavenging, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP)) and antidiabetic potential (Glycemic index (GI) and inhibition activity of α-amylase, and α-glucosidase) of species. The antioxidant activity increased 20% in CA3, 14% in CA1, and 8% in CA2. Furthermore, PCA showed significant variations not only between Mo-treated and untreated samples but also among Canavalia species. Overall, this study indicated that the sprouts of Canavalia species have tremendous potential for commercial usage due to their high nutritive value, which can be enhanced further with Mo treatment to accomplish the demand for nutritious feed.
ABSTRACT
N-acetyl-5-neuraminic acid (NeuAc) plays crucial role in improving the growth, brain development, brain health maintenance, and immunity enhancement of infants. Commercially, it is used in the production of antiviral drugs, infant milk formulas, cosmetics, dietary supplements, and pharmaceutical products. Because of the rapidly increasing demand, metabolic engineering approach has attracted increasing attention for NeuAc biosynthesis. However, knowledge of metabolite flux in biosynthetic pathways is one of the major challenges in the practice of metabolic engineering. So, an understanding of the flux of NeuAc is needed to determine its cellular level at real time. The analysis of the flux can only be performed using a tool that has the capacity to measure metabolite level in cells without affecting other metabolic processes. A Fluorescence Resonance Energy Transfer (FRET)-based genetically-encoded nanosensor has been generated in this study to monitor the level of NeuAc in prokaryotic and eukaryotic cells. Sialic acid periplasmic binding protein (SiaP) from Haemophilus influenzae was exploited as a sensory element for the generation of nanosensor. The enhanced cyan fluorescent protein (ECFP) and Venus were used as Fluroscence Resonance Energy Transfer (FRET) pair. The nanosensor, which was termed fluorescent indicator protein for sialic acid (FLIP-SA), was successfully transformed into, and expressed in Escherichia coli BL21 (DE3) cells. The expressed protein of the nanosensor was isolated and purified. The purified nanosensor protein was characterized to assess the affinity, specificity, and stability in the pH range. The developed nanosensor exhibited FRET change after addition to NeuAc. The developed nanosensor was highly specific, exhibited pH stability, and detected NeuAc levels in the nanomolar to milimolar range. FLIP-SA was successfully introduced in bacterial and yeast cells and reported the real-time intracellular levels of NeuAc non-invasively. The FLIP-SA is an excellent tool for the metabolic flux analysis of the NeuAc biosynthetic pathway and, thus, may help unravel the regulatory mechanism of the metabolic pathway of NeuAc. Furthermore, FLIP-SA can be used for the high-throughput screening of E. coli mutant libraries for varied NeuAc production levels.
ABSTRACT
Vitamin E plays an exemplary role in living organisms. α-Tocopherol is the most superior and active form of naturally occurring vitamin E that meets the requirements of human beings as it possesses the α-tocopherol transfer protein (α-TTP). α-Tocopherol deficiency can lead to severe anemia, certain cancers, several neurodegenerative and cardiovascular diseases, and most importantly male infertility. As a result of the depletion of its natural sources, researchers have tried to employ metabolic engineering to enhance α-tocopherol production to meet the human consumption demand. However, the metabolic engineering approach relies on the metabolic flux of a metabolite in its biosynthetic pathway. Analysis of the metabolic flux of a metabolite needs a method that can monitor the α-tocopherol level in living cells. This study was undertaken to construct a FRET (fluorescence resonance energy transfer)-based nanosensor for monitoring the α-tocopherol flux in prokaryotic and eukaryotic living cells. The human α-TTP was sandwiched between a pair of FRET fluorophores to construct the nanosensor, which was denoted as FLIP-α (the fluorescence indicator for α-tocopherol). FLIP-α showed excellence in monitoring the α-tocopherol flux with high specificity. The sensor was examined for its pH stability for physiological applications, where it shows no pH hindrance to its activity. The calculated affinity of this nanosensor was 100 µM. It monitored the real-time flux of α-tocopherol in bacterial and yeast cells, proving its biocompatibility in monitoring the α-tocopherol dynamics in living cells. Being noninvasive, FLIP-α provides high temporal and spatial resolutions, which holds an indispensable significance in bioimaging metabolic pathways that are highly compartmentalized.
ABSTRACT
Increased problems associated with side effects and bacterial resistance of chemical drugs has prompted the research focus on herbal medicines in the past few decades. In the present investigation, the antimicrobial activity of the various parts of Avicennia marina (AM), a mangrove plant, has been evaluated. The plants were collected from the Jazan area of the Kingdom of Saudi Arabia. Primary extracts of roots, stem, leaves, fruits, and seeds were made in ethanol and fractioned in ethanol, ethyl acetate, petroleum ether, chloroform, and water. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extracts were determined against Bacillussubtilis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. It has been observed that the chloroform extract of roots of the AM exhibited inhibitory effects against both S. aureus (MIC = 1.5 ± 0.03 mg/mL) and E. coli (MIC = 1.7 ± 0.01 mg/mL). The ethanolic extract of the AM roots has shown antibacterial activity against Pseudomonas aeruginosa (MIC = 10.8 ± 0.78 mg/mL), Bacillussubtilis (MIC = 6.1 ± 0.27 mg/mL), Staphylococcus aureus (MIC = 2.3 ± 0.08 mg/mL), and Escherichia coli (MIC = 6.3 ± 0.28 mg/mL). The leaf extract of the AM in ethyl acetate showed antibacterial activity against S. aureus and E. coli. Antifungal activity of these extracts was also investigated against Aspergillus fumigatus and Candida albicans. Ethanolic extract of roots and seeds of the AM has shown antifungal activity against Aspergillus fumigatus when applied individually. Ethanolic extract of the AM fruits has shown an inhibitory effect on the growth of Aspergillus fumigatus and Candida albicans. It is suggested that the plant extracts of AM have tremendous antimicrobial activity against a group of microbes, and this effect depends on both the plant part and the solvent used for extraction. Therefore, this plant can be considered to treat various diseases caused by antibiotic-resistant bacteria.
ABSTRACT
Colorimetric nanocomposite film sensor was fabricated by incorporating TCFH spectroscopic probe into cellulose nanowhiskers (CNW)/Urease enzyme matrix. CNW-TCFH can be used as disposable molecular biosensor in which CNW is the probe carrier comprising high surface area-to-volume ratio, urease is the catalyst and TCFH is the molecular probe. Tricyanofuran-hydrazone (TCFH) spectroscopic probe was prepared. UV-vis absorption spectra demonstrated solvatochromic behavior and a reversible color change of the tricyanofuran-hydrazone probe solution in acetone under acid/base conditions. CNW were reinforced with sodium alginate biopolymer to introduce biocomposite film. This CNW-TCFH film biosensor responds through visible color shift from light yellow to pink when exposed to urea in aqueous media. The morphology properties of CNW and CNW-TCFH films were examined by different tools. The photophysical properties of the prepared TCFH probe, including solvatochromism and pH sensory, were also studied.
