Search results for: antioxidant enzymes

Authors: Zoi Konsoula

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The increasing prevalence of Alzheimer’s disease (AD) and diabetes mellitus (DM) constitutes them major global health problems. Recently, the inhibition of key enzyme activity is considered a potential treatment of both diseases. Specifically, inhibition of acetylcholinesterase (AChE), the key enzyme involved in the breakdown of the neurotransmitter acetylcholine, is a promising approach for the treatment of AD, while inhibition of α-amylase retards the hydrolysis of carbohydrates and, thus, reduces hyperglycemia. Unfortunately, commercially available AChE and α-amylase inhibitors are reported to possess side effects. Consequently, there is a need to develop safe and effective treatments for both diseases. In the present study, pomegranate peel (PP) was extracted using various solvents of increasing polarity, while two extraction methods were employed, the conventional maceration and the ultrasound assisted extraction (UAE). The concentration of bioactive phytoconstituents, such as total phenolics (TPC) and total flavonoids (TFC) in the prepared extracts was evaluated by the Folin-Ciocalteu and the aluminum-flavonoid complex method, respectively. Furthermore, the anti-neurodegenerative and anti-hyperglycemic activity of all extracts was determined using AChE and α-amylase inhibitory activity assays, respectively. The inhibitory activity of the extracts against AChE and α-amylase was characterized by estimating their IC₅₀ value using a dose-response curve, while galanthamine and acarbose were used as positive controls, respectively. Finally, the kinetics of AChE and α-amylase in the presence of the most inhibitory potent extracts was determined by the Lineweaver-Burk plot. The methanolic extract prepared using the UAE contained the highest amount of phytoconstituents, followed by the respective ethanolic extract. All extracts inhibited acetylcholinesterase in a dose-dependent manner, while the increased anticholinesterase activity of the methanolic (IC₅₀ = 32 μg/mL) and ethanolic (IC₅₀ = 42 μg/mL) extract was positively correlated with their TPC content. Furthermore, the activity of the aforementioned extracts was comparable to galanthamine. Similar results were obtained in the case of α-amylase, however, all extracts showed lower inhibitory effect on the carbohydrate hydrolyzing enzyme than on AChE, since the IC₅₀ value ranged from 84 to 100 μg/mL. Also, the α-amylase inhibitory effect of the extracts was lower than acarbose. Finally, the methanolic and ethanolic extracts prepared by UAE inhibited both enzymes in a mixed (competitive/noncompetitive) manner since the Kₘ value of both enzymes increased in the presence of extracts, while the Vmax value decreased. The results of the present study indicate that PP may be a useful source of active compounds for the management of AD and DM. Moreover, taking into consideration that PP is an agro-industrial waste product, its valorization could not only result in economic efficiency but also reduce the environmental pollution.

Keywords: acetylcholinesterase, Alzheimer’s disease, α-amylase, diabetes mellitus, pomegranate

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Authors: Fadila Mohamed Mahmoud., Derkaoui I., Krimi Z.

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Date palm is a plant which presents a very good adaptation to the difficult conditions of the environment in particular to the drought and saline stress even at high temperatures. This adaptation is related on the biology of the plant and to the presence of a microflora endophyte which live inside its tissues. Fifteen endophytics fungi isolated from date palm were tested in vitro in the presence of various NaCl concentrations to select halotolerantes isolates. These same endophytes were tested for their colonizing capacity by the description of the production of secondary metabolites more particularly the enzymes (pectinases, proteases, and phosphorylases), and the production of antibiotics and growth hormones. Significant difference was observed between the isolates with respect to the tests carried out.

Keywords: Date palm, Halotolerantes, endophyte, Secondary metabolites.

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Authors: Fuad Ameen, Mohamed Moslem, Sarfaraz Hadi

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Twenty-eight fungal isolates belonging to 12 genera were derived from debris, sediment and water samples collected from Avicennia marina stands 25km south of Jeddah city on the Red Sea coast of Saudi Arabia. Eight of these isolates were found to be able to grow in association cellulosic waste materials under in vitro conditions in the absence of any carbon source. Isolates were further tested for their potential to degrade paper and clothes wastes by co-cultivation under aeration on a rotary shaker. These fungi accumulated significantly higher biomass, produced ligninolytic and cellulase enzymes, and liberated larger volumes of CO2. These observations indicated that the selected isolates were able to break down and consume the waste materials.

Keywords: biodegradation, enzyme activity, waste materials, mangrove

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Authors: Niki Mougiou, Spyros Didos, Ioanna Bouzouka, Athina Theodorakopoulou, Michael Kornaros, Anagnostis Argiriou

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Grains from traditional old Greek cereal varieties were evaluated and compared to commercial cultivars, like Simeto and Mexicali 81, in an effort to valorize local products and assess the nutritional benefits of ancient grains. The samples studied in this research included common wheat, durum wheat, emmer (Triticum dicoccum) and einkorn (Triticum monococcum), as well as barley, oats and rye grains. The Internal Transcribed Spacer 2 (ITS2) nuclear region was amplified and sequenced as a barcode for species identification, allowing the verification of the label of each product. After that, the total content of bound and free polyphenols and flavonoids, as well as the antioxidant activity of bound and free compounds, was measured by classic colorimetric assays using Folin- Ciocalteu, AlCl₃ and DPPH‧ (2,2-diphenyl-1-picrylhydrazyl) reagents, respectively. Moreover, the level of variation of fatty acids was determined in all samples by gas chromatography. The results showed that local old landraces of emmer and einkorn had the highest polyphenol content, 2.4 and 3.3 times higher than the average value of 5 durum wheat samples, respectively. Regarding the total flavonoid content, einkorn had 2.6-fold and emmer 2-fold higher values than common wheat. The antioxidant activity of free or bound compounds was at the same level, at about 20-30% higher in both einkorn and emmer compared to common wheat. Five main fatty acids were detected in all samples, in order of decreasing amounts: linoleic (C18:2) > palmitic (C16:0) ≈ , oleic (C18:1) > eicosenoic (C20:1, cis-11) > stearic (C18:0). Emmer and einkorn showed a higher diversity of fatty acids and a higher content of mono-unsaturated fatty acids compared to common wheat. The results of this study demonstrate the high nutritional value of old local landraces that have been put aside by more productive, yet with lower qualitative characteristics, commercial cultivars, underlining the importance of maintaining sustainable agricultural practices to ensure their continued cultivation.

Keywords: biochemical analysis, nutritional value, plant barcoding, wheat

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Authors: Rajeev Ravindran, Amit K. Jaiswal

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Lignocellulose is the largest reservoir of inexpensive, renewable source of carbon. It is composed of lignin, cellulose and hemicellulose. Cellulose and hemicellulose is composed of reducing sugars glucose, xylose and several other monosaccharides which can be metabolised by microorganisms to produce several value added products such as biofuels, enzymes, aminoacids etc. Enzymatic treatment of lignocellulose leads to the release of monosaccharides such as glucose and xylose. However, factors such as the presence of lignin, crystalline cellulose, acetyl groups, pectin etc. contributes to recalcitrance restricting the effective enzymatic hydrolysis of cellulose and hemicellulose. In order to overcome these problems, pre-treatment of lignocellulose is generally carried out which essentially facilitate better degradation of lignocellulose. A range of pre-treatment strategy is commonly employed based on its mode of action viz. physical, chemical, biological and physico-chemical. However, existing pretreatment strategies result in lower sugar yield and formation of inhibitory compounds. In order to overcome these problems, we proposes a novel pre-treatment, which utilises the superior oxidising capacity of alkaline potassium permanganate assisted by ultra-sonication to break the covalent bonds in spent coffee waste to remove recalcitrant compounds such as lignin. The pre-treatment was conducted for 30 minutes using 2% (w/v) potassium permanganate at room temperature with solid to liquid ratio of 1:10. The pre-treated spent coffee waste (SCW) was subjected to enzymatic hydrolysis using enzymes cellulase and hemicellulase. Shake flask experiments were conducted with a working volume of 50mL buffer containing 1% substrate. The results showed that the novel pre-treatment strategy yielded 7 g/L of reducing sugar as compared to 3.71 g/L obtained from biomass that had undergone dilute acid hydrolysis after 24 hours. From the results obtained it is fairly certain that ultrasonication assists the oxidation of recalcitrant components in lignocellulose by potassium permanganate. Enzyme hydrolysis studies suggest that ultrasound assisted alkaline potassium permanganate pre-treatment is far superior over treatment by dilute acid. Furthermore, SEM, XRD and FTIR were carried out to analyse the effect of the new pre-treatment strategy on structure and crystallinity of pre-treated spent coffee wastes. This novel one-step pre-treatment strategy was implemented under mild conditions and exhibited high efficiency in the enzymatic hydrolysis of spent coffee waste. Further study and scale up is in progress in order to realise future industrial applications.

Keywords: spent coffee waste, alkaline potassium permanganate, ultra-sonication, physical characterisation

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Authors: Camelia Vizireanu, Daniela Istrati, Alina Georgiana Profir, Rodica Mihaela Dinica

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Globally, there is a great interest in promoting the consumption of fruit and vegetables to improve health. Due to the content of essential compounds such as antioxidants, important amounts of fruits and vegetables should be included in the daily diet. Juices are good sources of vitamins and can also help increase overall fruit and vegetable consumption. Starting from this trend (introduction into the daily diet of vegetables and fruits) as well as the desire to diversify the range of functional products for both adults and children, a fermented juice was made using probiotic microorganisms based on root vegetables, with potential beneficial effects in the diet of children, vegetarians and people with lactose intolerance. The three vegetables selected for this study, red beet, carrot, and celery bring a significant contribution to functional compounds such as carotenoids, flavonoids, betalain, vitamin B and C, minerals and fiber. By fermentation, the functional value of the vegetable juice increases due to the improved stability of these compounds. The combination of probiotic microorganisms and vegetable fibers resulted in a nutrient-rich synbiotic product. The stability of the nutritional and sensory qualities of the obtained synbiotic product has been tested throughout its shelf life. The evaluation of the physico-chemical changes of the synbiotic drink during storage confirmed that: (i) vegetable juice enriched with honey and vegetable pulp is an important source of nutritional compounds, especially carbohydrates and fiber; (ii) microwave treatment used to inhibit pathogenic microflora did not significantly affect nutritional compounds in vegetable juice, vitamin C concentration remained at baseline and beta-carotene concentration increased due to increased bioavailability; (iii) fermentation has improved the nutritional quality of vegetable juice by increasing the content of B vitamins, polyphenols and flavonoids and has a good antioxidant capacity throughout the shelf life; (iv) the FTIR and Raman spectra have highlighted the results obtained using physicochemical methods. Based on the analysis of IR absorption frequencies, the most striking bands belong to the frequencies 3330 cm⁻¹, 1636 cm⁻¹ and 1050 cm⁻¹, specific for groups of compounds such as polyphenols, carbohydrates, fatty acids, and proteins. Statistical data processing revealed a good correlation between the content of flavonoids, betalain, β-carotene, ascorbic acid and polyphenols, the fermented juice having a stable antioxidant activity. Also, principal components analysis showed that there was a negative correlation between the evolution of the concentration of B vitamins and antioxidant activity. Acknowledgment: This study has been founded by the Francophone University Agency, Project Réseau régional dans le domaine de la santé, la nutrition et la sécurité alimentaire (SaIN), No. at Dunarea de Jos University of Galati 21899/ 06.09.2017 and by the Sectorial Operational Programme Human Resources Development of the Romanian Ministry of Education, Research, Youth and Sports trough the Financial Agreement POSDRU/159/1.5/S/132397 ExcelDOC.

Keywords: bioactive compounds, fermentation, synbiotic drink from vegetables, stability during storage

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Authors: C. C. Castro, T. Senechal, D. Lahem, A. L. Hantson

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Formaldehyde is one of the most representative volatile organic compounds present in the indoor air of residential units and workplaces. Increased attention has been given to this toxic compound because of its carcinogenic effect in health. Biological or enzymatic transformation is being explored to degrade this pollutant. Pseudomonas putida is a bacteria able to synthesize formaldehyde dehydrogenase, an enzyme known to use formaldehyde as a substrate and transform it into less toxic compounds. The immobilization of bacterial cells in the surface of different supports through spraying or dip-coating is herein proposed. The determination of the enzymatic activity on the coated surfaces was performed as well as the study of its effect on formaldehyde degradation in an isolated chamber. Results show that the incorporation of microbial cells able to synthesize depolluting enzymes can be an innovative, low-cost, effective and environmentally friendly solution for indoor air depollution.

Keywords: cells encapsulation, formaldehyde, formaldehyde dehydrogenase, indoor air depollution

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Authors: Zlatina Genisheva, Pedro Ferreira-Santos, Margarida Soares, Cândida Vilarinho, Joana Carvalho

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The wine industry produces significant quantities of waste, much of which remains underutilized as a potential raw material. Typically, this waste is either discarded in the fields or incinerated, leading to environmental concerns. By-products of wine production, like lees and grape pomace, are readily available at relatively low costs and hold promise as raw materials for biochemical conversion into valuable products. Reusing these waste materials is crucial, not only for reducing environmental impact but also for enhancing profitability. The Vinhos Verdes demarcated region, the largest wine-producing area in Portugal, has remained relatively stagnant over time. This project aims to offer an alternative income source for producers in the region while also expanding the limited existing research on this area. The main objective of this project is the study of the sustainable valorization of grape pomace and lees from the production of DOC Vinho Verde. Extraction tests were performed to obtain high-value compounds, targeting phenolic compounds from grape pomace and protein-rich extracts from lees. An environmentally friendly technique, microwave extraction, was used for this process. This method is not only efficient but also aligns with the principles of green chemistry, reducing the use of harmful solvents and minimizing energy consumption. The findings from this study have the potential to open new revenue streams for the region’s wine producers while promoting environmental sustainability. The optimal conditions for extracting proteins from lees involve the use of NaOH at 150ºC. Regardless of the solvent employed, the ideal temperature for obtaining extracts rich in polyphenol compounds and exhibiting strong antioxidant activity is also 150ºC. For grape pomace, extracts with a high concentration of polyphenols and significant antioxidant properties were obtained at 210ºC. However, the highest total tannin concentrations were achieved at 150ºC, while the maximum total flavonoid content was obtained at 170ºC.

Keywords: antioxidants, circular economy, polyphenol compounds, waste valorization

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Authors: Mesfin Angaw Tesfay

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Uridine-5’-diphosphate (UDP)-glucose is one of the most versatile building blocks within the metabolism of prokaryotes and eukaryotes, serving as an activated sugar donor during the glycosylation of natural products. It is formed by the enzyme UDP-glucose pyrophosphorylase (UGPase) using uridine-5′-triphosphate (UTP) and α-d-glucose 1-phosphate as a substrate. Herein, two UGPase genes from Lactobacillus kefiranofaciens ZW3 (LkUGPase) and Rhodococcus wratislaviensis IFP 2016 (RwUGPase) were identified through genome mining approaches. The LkUGPase and RwUGPase have 299 and 306 amino acids, respectively. Both UGPase has the conserved UTP binding site (G-X-G-T-R-X-L-P) and the glucose -1-phosphate binding site (V-E-K-P). The LkUGPase and RwUGPase were cloned in E. coli, and SDS-PAGE analysis showed the expression of both enzymes forming about 36 KDa of protein band after induction. LkUGPase and RwUGPase have an activity of 1549.95 and 671.53 U/mg, respectively. Currently, their kinetic properties are under investigation.

Keywords: UGPase, LkUGPase, RwUGPase, UDP-glucose, glycosylation

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Authors: Mirjana Petronijević, Sanja Panić, Aleksandra Cvetanović, Branko Kordić, Nenad Grba

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Enzyme peroxidases are biological catalysts and play a major role in phenolic wastewater treatments and other environmental applications. The most studied species from the peroxidases family is horseradish peroxidase (HRP). In environmental processes, HRP could be used in its free or immobilized form. Enzyme immobilization onto solid support is performed to improve the enzyme properties, prolong its lifespan and operational stability and allow its reuse in industrial applications. One of the enzyme supports of a newer generation is magnetic particles (MPs). Fe₃O₄ MPs are the most widely pursued immobilization of enzymes owing to their remarkable advantages of biocompatibility and non-toxicity. Also, MPs can be easily separated and recovered from the water by applying an external magnetic field. On the other hand, metals and metal oxides are not suitable for the covalent binding of enzymes, so it is necessary to perform their surface modification. Fe₃O₄ MPs functionalization could be performed during the process of their synthesis if it takes place in the presence of plant extracts. Extracts of plant material, such as wild plants, herbs, even waste materials of the food and agricultural industry (bark, shell, leaves, peel), are rich in various bioactive components such as polyphenols, flavonoids, sugars, etc. When the synthesis of magnetite is performed in the presence of plant extracts, bioactive components are incorporated into the surface of the magnetite, thereby affecting its functionalization. In this paper, the suitability of bio-magnetite as solid support for covalent immobilization of HRP across glutaraldehyde was examined. The activity of immobilized HRP at different pH values (4-9) and temperatures (20-80°C) and reusability were examined. Bio-MP was synthesized by co-precipitation method from Fe(II) and Fe(III) sulfate salts in the presence of water extract of the Allium cepa peel. The water extract showed 81% of antiradical potential (according to DPPH assay), which is connected with the high content of polyphenols. According to the FTIR analysis, the bio-magnetite contains oxygen functional groups (-OH, -COOH, C=O) suitable for binding to glutaraldehyde, after which the enzyme is covalently immobilized. The immobilized enzyme showed high activity at ambient temperature and pH 7 (30 U/g) and retained ≥ 80% of its activity at a wide range of pH (5-8) and temperature (20-50°C). The HRP immobilized onto bio-MPs showed remarkable stability towards temperature and pH variations compared to the free enzyme form. On the other hand, immobilized HRP showed low reusability after the first washing cycle enzyme retains 50% of its activity, while after the third washing cycle retains only 22%.

Keywords: bio-magnetite, enzyme immobilization, water extracts, environmental protection

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Authors: F. Pires, V. Geraldo, O. N. Oliveira Jr., M. Raposo

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Catechins are powerful antioxidants which have attractive properties useful for tumor therapy. Considering their antioxidant activity, these molecules can act as a scavenger of the reactive oxygen species (ROS), alleviating the damage of cell membrane induced by oxidative stress. The complexity and dynamic nature of the cell membrane compromise the analysis of the biophysical interactions between drug and cell membrane and restricts the transport or uptake of the drug by intracellular targets. To avoid the cell membrane complexity, we used biomimetic systems as liposomes and Langmuir monolayers to study the interaction between catechin and membranes at the molecular level. Liposomes were formed after the dispersion of anionic 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)(sodium salt) (DPPG) phospholipids in an aqueous solution, which mimic the arrangement of lipids in natural cell membranes and allows the entrapment of catechins. Langmuir monolayers were formed after dropping amphiphilic molecules, DPPG phospholipids, dissolved in an organic solvent onto the water surface. In this work, we mixed epigallocatechin-3-gallate (EGCG) with DPPG liposomes and exposed them to ultra-violet radiation in order to evaluate the antioxidant potential of these molecules against oxidative stress induced by radiation. The presence of EGCG in the mixture decreased the rate of lipid peroxidation, proving that EGCG protects membranes through the quenching of the reactive oxygen species. Considering the high amount of hydroxyl groups (OH groups) on structure of EGCG, a possible mechanism to these molecules interact with membrane is through hydrogen bonding. We also investigated the effect of EGCG at various concentrations on DPPG Langmuir monolayers. The surface pressure isotherms and infrared reflection-absorption spectroscopy (PM-IRRAS) results corroborate with absorbance results preformed on liposome-model, showing that EGCG interacts with polar heads of the monolayers. This study elucidates the physiological action of EGCG which can be incorporated in lipid membrane. These results are also relevant for the improvement of the current protocols used to incorporate catechins in drug delivery systems.

Keywords: catechins, lipid membrane, anticancer agent, molecular interactions

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

Abstract:

One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

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Authors: Callistus I. Iheme, Kenneth E. Asika, Emmanuel I. Ugwor, Chukwuka U. Ogbonna, Ugonna H. Uzoka, Nneamaka A. Chiegboka, Chinwe S. Alisi, Obinna S. Nwabueze, Amanda U. Ezirim, Judeanthony N. Ogbulie

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Antimicrobial resistance (AMR) is a major threat to the global health sector. Zinc oxide nanocomposites (ZnONCs), composed of zinc oxide nanoparticles and phytochemicals from Azadirachta indica aqueous leaf extract, were assessed for their physico-chemicals, in silico and in vitro antimicrobial properties on multidrug-resistant Escherichia coli enzymes. Gas chromatography coupled with mass spectroscope (GC-MS) analysis on the ZnONCs revealed the presence of twenty volatile phytochemical compounds, among which is scoparone. Characterization of the ZnONCs was done using ultraviolet-visible spectroscopy (UV-vis), energy dispersive spectroscopy (EDX), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and x-ray diffractometer (XRD). Dehydrogenase enzyme converts colorless 2,3,5-triphenyltetrazolium chloride to the red triphenyl formazan (TPF). The rate of formazan formation in the presence of ZnONCs is proportional to the enzyme activities. The color formation is extracted and determined at 500 nm, and the percentage of enzyme activity is calculated. To determine the bioactive components of the ZnONCs, characterize their binding to enzymes, and evaluate the enzyme-ligand complex stability, respectively Discrete Fourier Transform (DFT) analysis, docking, and molecular dynamics simulations will be employed. The results showed arrays of ZnONCs nanorods with maximal absorption wavelengths of 320 nm and 350 nm and thermally stable at the temperature range of 423.77 to 889.69 ℃. In vitro study assessed the dehydrogenase inhibitory properties of the ZnONCs, conjugate of ZnONCs and ampicillin (ZnONCs-amp), the aqueous leaf extract of A. indica, and ampicillin (standard drug). The findings revealed that at the concentration of 500 μm/mL, 57.89 % of the enzyme activities were inhibited by ZnONCs compared to 33.33% and 21.05% of the standard drug (Ampicillin), and the aqueous leaf extract of the A. indica respectively. The inhibition of the enzyme activities by the ZnONCs at 500 μm/mL was further enhanced to 89.74 % by conjugating with Ampicillin. In silico study on the ZnONCs revealed scoparone as the most viable competitor of nicotinamide adenine dinucleotide (NAD⁺) for the coenzyme binding pocket on E. coli malate and histidinol dehydrogenase. From the findings, it can be concluded that the scoparone components of the nanocomposites in synergy with the zinc oxide nanoparticles inhibited E. coli malate and histidinol dehydrogenase by competitively binding to the NAD⁺ pocket and that the conjugation of the ZnONCs with ampicillin further enhanced the antimicrobial efficiency of the nanocomposite against multidrug resistant E. coli.

Keywords: antimicrobial resistance, dehydrogenase activities, E. coli, zinc oxide nanocomposites

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Authors: Temesgen Geremew Tefery

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Nanomaterials, due to their unique properties, have revolutionized the field of biosensing. Their functionalization, or modification with specific molecules, is crucial for enhancing their biocompatibility, selectivity, and sensitivity. This review explores recent advancements in nanomaterial functionalization for biosensing applications. We discuss various strategies, including covalent and non-covalent modifications, and their impact on biosensor performance. The use of biomolecules like antibodies, enzymes, and nucleic acids for targeted detection is highlighted. Furthermore, the integration of nanomaterials with different sensing modalities, such as electrochemical, optical, and mechanical, is examined. The future outlook for nanomaterial-based biosensing is promising, with potential applications in healthcare, environmental monitoring, and food safety. However, challenges related to biocompatibility, scalability, and cost-effectiveness need to be addressed. Continued research and development in this area will likely lead to even more sophisticated and versatile biosensing technologies.

Keywords: biosensing, nanomaterials, biotechnology, nanotechnology

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Authors: H. Y. Al-Matubsi, G. A. Oriquat, M. Abu-Samak, O. A. Al Hanbali, M. Salim

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Background: Uncontrolled diabetes mellitus (DM) is an etiological factor for recurrent pregnancy loss and major congenital malformations in the offspring. Antioxidant therapy has been advocated to overcome the oxidant-antioxidant disequilibrium inherent in diabetes. The aims of this study were to evaluate the protective effect of lipoic acid (LA) on fetal outcome and to elucidate changes that may be involved in the mechanism(s) implicit diabetic fetopathy. Methods: Female rats were rendered hyperglycemic using streptozocin and then mated with normal male rat. Pregnant non-diabetic (group1; n=9; and group2; n=7) or pregnant diabetic (group 3; n=10; and group 4; n=8) rats were treated daily with either lipoic acid (LA) (30 mg/kg body weight; groups 2 and 4) or vehicle (groups 1 and 3) between gestational days 0 and 15. On day 15 of gestation, the rats were sacrificed, and the fetuses, placentas and membranes dissected out of the uterine horns. Following morphological examination, the fetuses, placentas and membranes were homogenized, and used to measure cyclooxygenases (COX) activities and metabolisms of prostaglandin (PG) E2 (PGEM) and PGF2 (PGFM) levels. Maternal liver and plasma total glutathione levels were also determined. Results: Supplementation of diabetic rats with LA was found to significantly (P<0.05) reduce resorption rates in diabetic rats and increased mean fetal weight compared to diabetic group. Treatment of diabetic rats with LA leads to a significant (P<0.05) increase in liver and plasma total glutathione, in comparison with diabetic rats. Decreased levels of PGEM and elevated levels of PGFM in the fetuses, placentas and membranes were characteristic of experimental diabetic gestation associated with malformation. LA treatment to diabetic mothers failed to normalize levels of PGEM to the non-diabetic control rats. However, the levels of PGEM in malformed fetuses from LA-treated diabetic mothers was significantly (P < 0.05) higher than those in malformed fetuses from diabetic rats. Conclusions: We conclude that LA can reduce congenital malformations in the offspring of diabetic rats at day 15 of gestation. However, LA treatment did not completely prevent the occurrence of malformations, other factors, such as arachidonic acid deficiency and altered prostaglandin metabolismmay be involved in the pathogenesis of diabetes-induced congenital malformations.

Keywords: diabetes, lipoic acid, pregnancy, prostaglandins

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Authors: Haniza Ahmad, Norliza Saparin, Ahmadilfitri Md Noor, Mohd Suria Affandi Yusoff

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Enzymatic remediation is applied in low free fatty acid (FFA) (<4%) crude palm oil (CPO) to investigate if further FFA reduction is able to take place to produce premium CPO (<1% FFA). There are four different lipase Candida Antartica brands used in this study. Samples submit to enzymatic remediation using rotary evaporator under 100mbar vacuum with rotation at 260rpm. Samples were taken at 4hours, 8hours and 24hours for analyses. FFA less than 1% was achieved after 24hours reaction with 1% enzyme and 2% glycerol. The FFA reduction was intensified with the presence of glycerol who provides more sites for fatty acid attachment. At 2% glycerol, 71-88% FFA was reduced whereas at 1% glycerol, 46-75% FFA reduced. However, partial glycerides was increased with presence of glycerol with 2% add in glycerol showed greater partial glycerides increment compared to 1% glycerol.

Keywords: enzymes, crude palm oil, free fatty acid, glycerol

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Authors: Mesfin Angaw Tesfay

Abstract:

Uridine-5’-diphosphate (UDP)-glucose is one of the most versatile building blocks within the metabolism of prokaryotes and eukaryotes serving as an activated sugar donor during the glycosylation of natural products. It is formed by the enzyme UDP-glucose pyrophosphorylase (UGPase) using uridine-5′-triphosphate (UTP) and α-d-glucose 1-phosphate as a substrate. Herein two UGPase genes from Lactobacillus kefiranofaciens ZW3 (LkUGPase) and Rhodococcus wratislaviensis IFP 2016 (RwUGPase) were identified through genome mining approaches. The LkUGPase and RwUGPase have 299 and 306 amino acids, respectively. Both UGPase has the conserved UTP binding site (G-X-G-T-R-X-L-P) and the glucose -1-phosphate binding site (V-E-K-P). The LkUGPase and RwUGPase were cloned in E. coli and SDS-PAGE analysis showed the expression of both enzymes forming about 36 KDa of protein band after induction. LkUGPase and RwUGPase have an activity of 1549.95 and 671.53 U/mg respectively. Currently, their kinetic properties are under investigation.

Keywords: UGPase, LkUGPase, RwUGPase, UDP-glucose, Glycosylation

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Authors: Mohammadhosein Rahimi, Mohammad Raouf Hosseini, Mehran Bakhshi, Alireza Baghbanan

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Silver ions (Ag⁺) and their compounds are consequentially toxic to microorganisms, showing biocidal effects on many species of bacteria. Silver-phosphate (or silver orthophosphate) is one of these compounds, which is famous for its antimicrobial effect and catalysis application. In the present study, a green method was presented to synthesis silver-phosphate nanoparticles using Sporosarcina pasteurii. The composition of the biosynthesized nanoparticles was identified as Ag₃PO₄ using X-ray Diffraction (XRD) and Energy Dispersive Spectroscopy (EDS). Also, Fourier Transform Infrared (FTIR) spectroscopy showed that Ag₃PO₄ nanoparticles was synthesized in the presence of biosurfactants, enzymes, and proteins. In addition, UV-Vis adsorption of the produced colloidal suspension approved the results of XRD and FTIR analyses. Finally, Transmission Electron Microscope (TEM) images indicated that the size of the nanoparticles was about 20 nm.

Keywords: bacteria, biosynthesis, silver-phosphate, Sporosarcina pasteurii, nanoparticle

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Authors: M. Akoz, O. B. Citil, I. Aydin

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Colostrum secreted by the mammary glands after birth in the early days, a high proportion of fat, protein and ash containing a secretion containing low amounts of casein and lactose. Especially immunoglobulins contain high proportions. Maternal immunoglobulins own immune system to protect the newborn against neonatal disease until development are very important matter. However, colostrum is transferred to the offspring due to placental barrier in ruminants. Immunoglobulins are absorbed through the intestinal epithelium but absorption can vary under the influence of some factors. These factors are among the priority ones taking colostrum first time, amount, concentration, the metabolic status of the newborn. intestinal absorption of immunoglobulins occurs over the first 24 h high. Absorption from the gut after nine hours, 50% after 24 hours was only 11%. On the other hand pup's digestive system degrade the enzymes after 24 hours immunoglobulins. Bovine colostrum in the composition while basic immune IgG, IgA and IgM are also available. Total IgG in colostrum of ruminants, while in other species is a greater amount in blood serum.

Keywords: immunoglobulin, ruminants, colostrum, immune system

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Authors: D. J. Madhu Kumar, D. Jagadeesh Prasad, Sana Sheik, E. P. Rejeesh

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A series of Mannich bases derived from 1,2,4-triazole(3a-k and 4a-k) are synthesized by treating a Schiff base with various substituted primary/secondary amines and formaldehyde. The Schiff base is prepared by treating 3-methyl-4-amino-5-mercapto-1,2,4-triazole with 3,4-dimethoxybenzaldehyde in the presence of acid catalyst. The triazole is prepared by treating acetic acid with thiocarbohydrazide at reflux temperature. All the synthesized samples are characterised by FT-IR, 1H-NMR, and LC-MASS spectral studies and screened for their anti-oxidant activity.

Keywords: mannich bases, anti-oxidant activity, schiff base, triazole

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Authors: Lazhar Zourgui

Abstract:

Opuntia species are used as local medicinal interventions for chronic diseases and as food sources, mainly because they possess nutritional properties and biological activities. Opuntia ficus-indica (L.) Mill, commonly known as prickly pear or nopal cactus, is the most economically valuable plant in the Cactaceae family worldwide. It is a tropical or subtropical plant native to tropical and subtropical America, which can grow in arid and semi-arid climates. It belongs to the family of angiosperms dicotyledons Cactaceae of which about 1500 species of cacti are known. The Opuntia plant is distributed throughout the world and has great economic potential. There are differences in the phytochemical composition of Opuntia species between wild and domesticated species and within the same species. It is an interesting source of plant bioactive compounds. Bioactive compounds are compounds with nutritional benefits and are generally classified into phenolic and non-phenolic compounds and pigments. Opuntia species are able to grow in almost all climates, for example, arid, temperate, and tropical climates, and their bioactive compound profiles change depending on the species, cultivar, and climatic conditions. Therefore, there is an opportunity for the discovery of new compounds from different Opuntia cultivars. Health benefits of prickly pear are widely demonstrated: There is ample evidence of the health benefits of consuming prickly pear due to its source of nutrients and vitamins and its antioxidant properties due to its content of bioactive compounds. In addition, prickly pear is used in the treatment of hyperglycemia and high cholesterol levels, and its consumption is linked to a lower incidence of coronary heart disease and certain types of cancer. It may be effective in insulin-independent type 2 diabetes mellitus. Opuntia ficus-Indica seed oil has shown potent antioxidant and prophylactic effects. Industrial applications of these bioactive compounds are increasing. In addition to their application in the pharmaceutical industries, bioactive compounds are used in the food industry for the production of nutraceuticals and new food formulations (juices, drinks, jams, sweeteners). In my lecture, I will review in a comprehensive way the phytochemical, nutritional, and bioactive compound composition of the different aerial and underground parts of Opuntia species. The biological activities and applications of Opuntia compounds are also discussed.

Keywords: medicinal plants, cactus, Opuntia, actives biomolecules, biological activities

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Authors: Houneïda Benbouzid, Houria Berrebbah, Mohammed-Réda Djebar

Abstract:

The toxicological impacts of the increasing number of synthetic compounds present in the aquatic environment are assessed predominantly in laboratory studies where test organisms are exposed to a range of concentrations of single compounds. The bio-indicator Paramecium sp., characterized by a short life cycle, rapid multiplication and normal behavior that may be affected by the presence of pollutants. We therefore investigated the inhibitory effect of a newly synthesized acaricide: the chlorfenapyr tested at concentrations of 250, 300, and 350 µM on a pure culture of Paramecium sp. during 6 day. Paramecia treated with different concentrations of Chlorfenapyr illustrate strong inhibition of cell growth from the second day of treatment. Low levels of glutathione, increased glutathione S-transferase and the decrease in respiratory metabolism, recorded in the presence of different concentrations of Chlorfenapyr, involve the activation of detoxification system.

Keywords: Paramecium sp., chlorfenapyr, oxidative enzymes, detoxification

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Authors: B. Selma, S. Chouraqui

Abstract:

The pancreas is an elongated organ that extends across the abdomen, below the stomach. In addition, it secretes certain enzymes that aid in food digestion. The pancreas also manufactures hormones responsible for regulating blood glucose levels. In the present paper, we propose a mathematical model to study the homeostasis of glucose and insulin in healthy human, and a simulation of this model, which depicts the physiological events after a meal, will be represented in ordinary humans. The aim of this paper is to design an algorithm which regulates the level of glucose in the blood. The algorithm applied the concept of expert system for performing an algorithm control in the form of an "active" used to prescribe the rate of insulin infusion. By decomposing the system into subsystems, we have developed parametric models of each subsystem by using a forcing function strategy. The results showed a performance of the control system.

Keywords: modeling, algorithm, regulation, glucose-insulin, blood, control system

Procedia PDF Downloads 175

Authors: P. Hashim, M. S. Mohd Ridzwan, J. Bakar

Abstract:

Collagen was isolated from chicken feet by using papain and pepsin enzymes in acetic acid solution at 4°C for 24h with a yield of 18.16% and 22.94% by dry weight, respectively. Chemical composition and characteristics of chicken feet collagen such as amino acid composition, SDS-PAGE patterns, FTIR spectra and thermal properties were evaluated. The chicken feet collagen is rich in the amino acids glycine, glutamic acid, proline and hydroxyproline. Electrophoresis pattern demonstrated two distinct α-chains (α1 and α2) and β chain, indicating that type I collagen is a major component of chicken feet collagen. The thermal stability of collagen isolated by papain and pepsin revealed stable denaturation temperatures of 48.40 and 53.35°C, respectively. The FTIR spectra of both collagens were similar with amide regions in A, B, I, II, and III. The study demonstrated that chicken feet collagen using papain isolation method is possible as commercial alternative ingredient.

Keywords: chicken feet, collagen, papain, pepsin

Procedia PDF Downloads 422

Authors: Mohamed Rehan, Gamil E. Ibrahim, Mohamed S. Abdel-Aziz, Shaimaa R. Ibrahim, Tawfik A. Khattab

Abstract:

The utilization of natural products in finishing textiles toward multifunctional applications without side effects is an extremely motivating goal. Hibiscus sabdariffa usually has been used for many traditional medicine applications. To develop an additional use for Hibiscus sabdariffa, an extraction of bioactive compounds from Hibiscus sabdariffa followed by finishing on cellulosic fibers was designed to cleaner production of the value-added textiles fibers with multifunctional applications. The objective of this study is to explore, identify, and evaluate the bioactive compound extracted from Hibiscus sabdariffa by different solvent via ultrasonic technique as a potential eco-friendly agent for multifunctional cellulosic fabrics via two approaches. In the first approach, Hibiscus sabdariffa extract was used as a source of sustainable eco-friendly for simultaneous coloration and multi-finishing of cotton fabrics via in situ incorporations of nanoparticles (silver and metal oxide). In the second approach, the micro-capsulation of Hibiscus sabdariffa extracts was followed by coating onto cotton gauze to introduce multifunctional healthcare applications. The effect of the solvent type was accelerated by ultrasonic on the phytochemical, antioxidant, and volatile compounds of Hibiscus sabdariffa. The surface morphology and elemental content of the treated fabrics were explored using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), and energy-dispersive X-ray spectroscopy (EDX). The multifunctional properties of treated fabrics, including coloration, sensor properties and protective properties against pathogenic microorganisms and UV radiation as well as wound healing property were evaluated. The results showed that the water, as well as ethanol/water, was selected as a solvent for the extraction of natural compounds from Hibiscus Sabdariffa with high in extract yield, total phenolic contents, flavonoid contents, and antioxidant activity. These natural compounds were utilized to enhance cellulosic fibers functionalization by imparting faint/dark red color, antimicrobial against different organisms, and antioxidants as well as UV protection properties. The encapsulation of Hibiscus Sabdariffa extracts, as well as wound healing, is under consideration and evaluation. As a result, the current study presents a sustainable and eco-friendly approach to design cellulosic fabrics for multifunctional medical and healthcare applications.

Keywords: cellulosic fibers, Hibiscus sabdariffa extract, multifunctional application, nanoparticles

Procedia PDF Downloads 144

Authors: Maritsa Kurashvili, George Adamia, Tamar Ananiashvili, Lia Amiranasvili, Tamar Varazi, Marina Pruidze, Marlen Gordeziani, Gia Khatisashvili

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The goal of presented work is the development phytoremediation method targeted to cleaning environment polluted with organochlorine pesticides, based on joint application of plants and microorganisms. For this aim the selection of plants and microorganisms with corresponding capabilities towards three organochlorine pesticides (Lindane, DDT and PCP) has been carried out. The tolerance of plants to tested pesticides and induction degree of plant detoxification enzymes by these compounds have been used as main criteria for estimating the applicability of plants in proposed technology. Obtained results show that alfalfa, maize and soybean among tested six plant species have highest tolerance to pesticides. As a result of screening, more than 30 strains from genera Pseudomonas have been selected. As a result of GC analysis of incubation area, 11 active cultures for investigated pesticides are carefully chosen.

Keywords: DDT, Lindane, organochlorine pesticides, PCP, phytoremediation

Procedia PDF Downloads 312

Authors: Mohamed S. Sasi, Adel M. Mlitan, Abdulfattah M. Alkherraz

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This research project aims to investigate difference in relative rates concerning phosphoryl transfer relevant to biological catalysis of DNA and RNA in the pH-independent reactions. Activated Models of DNA and RNA for alkyl-aryl phosphate diesters (with 4-nitrophenyl as a good leaving group) have successfully been prepared to gather kinetic parameters. Eyring plots for the pH–independent hydrolysis of 1 and 2 were established at different temperatures in the range 100–160 °C. These measurements have been used to provide a better estimate for the difference in relative rates between the reactivity of DNA and RNA cleavage. Eyring plot gave an extrapolated rate of kH2O = 1 × 10-10 s -1 for 1 (RNA model) and 2 (DNA model) at 25°C. Comparing the reactivity of RNA model and DNA model shows that the difference in relative rates in the pH-independent reactions is surprisingly very similar at 25°. This allows us to obtain chemical insights into how biological catalysts such as enzymes may have evolved to perform their current functions.

Keywords: DNA and RNA models, relative rates, reactivity, phosphoryl transfe

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Authors: Nassima Khanfri, Ali Boucenna

Abstract:

As metallic nanoparticles are increasingly used in many economic sectors, there is interest in the biological and environmental safety of their production. The main methods of synthesizing nanoparticales are chemical and physical approaches that are often expensive and potentially harmful to the environment. The present study is devoted to the possibility of the synthesis of silver nanoparticales and zinc oxide from silver nitrate and zinc acetate using basilica plant extracts. The products obtained are characterized by various analysis techniques, such as UV/V, XRD, MEB-EDX, FTIR, and RAMAN. These analyzes confirm the crystalline nature of AgNps and ZnONps. These crystalline powders having effective biological activities regarding the antioxidant and antibacterial, which could be used in several biological applications.

Keywords: green synthesis, bio-reduction, metals nan Oparticales, Plants extracts

Procedia PDF Downloads 196

Authors: Anshu Siwach, Qianlai Zhuang, Ratul Baishya

Abstract:

Context: This study focuses on the influence of moss cover and seasonality on soil microbial biomass and enzymatic activity in different Central Himalayan temperate forest types. Soil microbial biomass and enzymes are key indicators of microbial communities in soil and provide information on soil properties, microbial status, and organic matter dynamics. The activity of microorganisms in the soil varies depending on the vegetation type and environmental conditions. Therefore, this study aims to assess the effects of moss cover, seasons, and different forest types on soil microbial biomass carbon (SMBC), soil microbial biomass nitrogen (SMBN), and soil enzymatic activity in the Central Himalayas, Uttarakhand, India. Research Aim: The aim of this study is to evaluate the levels of SMBC, SMBN, and soil enzymatic activity in different temperate forest types under the influence of two ground covers (soil with and without moss cover) during the rainy and winter seasons. Question Addressed: This study addresses the following questions: 1. How does the presence of moss cover and seasonality affect soil microbial biomass and enzymatic activity? 2. What is the influence of different forest types on SMBC, SMBN, and enzymatic activity? Methodology: Soil samples were collected from different forest types during the rainy and winter seasons. The study utilizes the chloroform-fumigation extraction method to determine SMBC and SMBN. Standard methodologies are followed to measure enzymatic activities, including dehydrogenase, acid phosphatase, aryl sulfatase, β-glucosidase, phenol oxidase, and urease. Findings: The study reveals significant variations in SMBC, SMBN, and enzymatic activity under different ground covers, within the rainy and winter seasons, and among the forest types. Moss cover positively influences SMBC and enzymatic activity during the rainy season, while soil without moss cover shows higher values during the winter season. Quercus-dominated forests, as well as Cupressus torulosa forests, exhibit higher levels of SMBC and enzymatic activity, while Pinus roxburghii forests show lower levels. Theoretical Importance: The findings highlight the importance of considering mosses in forest management plans to improve soil microbial diversity, enzymatic activity, soil quality, and health. Additionally, this research contributes to understanding the role of lower plants, such as mosses, in influencing ecosystem dynamics. Conclusion: The study concludes that moss cover during the rainy season significantly influences soil microbial biomass and enzymatic activity. Quercus and Cupressus torulosa dominated forests demonstrate higher levels of SMBC and enzymatic activity, indicating the importance of these forest types in sustaining soil microbial diversity and soil health. Including mosses in forest management plans can improve soil quality and overall ecosystem dynamics.

Keywords: moss cover, seasons, soil enzymes, soil microbial biomass, temperate forest types

Procedia PDF Downloads 63

Authors: Aqeel Ahmed Shad, Tanveer Ahmad, Muhammad Farooq Iqbal, Muhammad Javaid Asad

Abstract:

The supplemental effects of novel protease produced from Bacillus subtilis K-5 and beta-mannanase were evaluated on growth performance, carcass characteristics, nutrients digestibility, blood profile and intestinal morphometry of broilers fed guar meal (Cyamopsis tetragonoloba) based diets with reduced Crude Protein (CP), Essential Amino Acids (EAAs), and Metabolizable energy (ME) contents. One-day old Ross 308 broiler chicks (n=360) were randomly allotted to thirty six experimental units in a way that each of the nine dietary treatments received four replicates with ten birds per replicate. A control diet without guar meal (0GM) was formulated with standard nutrient specifications of Ross 308 for the starter and finisher phases. Two negative control diets, one with 5% (5GM) and second with 10% (10GM) guar meal, were formulated with reduction of 5% CP, 5% EAAs and 80 Kcal/kg ME. These three basal diets (no enzyme) were supplemented with novel protease enzyme (PROT) and commercial beta-mannanase (Beta-M) enzyme. The birds were reared up to 35d of age. The data on weekly body weight gain (BWG) and feed intake were recorded to compute feed:gain for the starter (0-21d) and finisher (22-35d) phases. At the end of 35d of experimental period, four birds per experimental unit were randomly selected for blood samples collection and later slaughtered for ileal digesta, intestinal tract and carcass trait sampling. The data on overall performance (1-35d) indicated improved (P<0.05) BWG and feed:gain in birds supplemented with PROT (1.41% and 1.67) and Beta-M (2.79% and 1.64) than non-supplemented groups. Improved (P<0.05) carcass yield, breast meat yield and thigh meat yield were noted with the supplementation of Beta-M. However, non-significant (P>0.05) effect on carcass traits was noted in broiler fed guar meal based PROT supplemented diets. Crude protein digestibility, nitrogen retention (Nret) and apparent digestibility coefficient for nitrogen (ADCN) were improved (P<0.05) only with PROT. The improvement in apparent metabolizable energy (AME) and apparent metabolizable energy corrected for nitrogen (AMEn) was noted (P<0.05) with both supplemented enzymes. However, no effect (P>0.05) of enzyme addition was noted on blood glucose, total protein and cholesterol. Improved villus height of duodenum, jejunum and ileum was noted (P<0.05) with the addition of both enzymes. The EAAs digestibility was improved (P<0.05) only with PROT. In conclusion, beta-mannanase and protease supplementation better improved the overall bird performance in low nutrient profile guar meal based diets than non-supplemented diets.

Keywords: novel protease, guar meal, broilers, low protein diets, low metabolizable energy diets, nutrients digestibility

Procedia PDF Downloads 58