Green ecofriendly enhancement of cellulase productivity using agricultural wastes by Aspergillus terreus MN901491: statistical designs and detergent ability on cotton fabrics.

BACKGROUND: Cellulase is considered a group member of the hydrolytic enzymes, responsible for catalyzing the hydrolysis of cellulose and has various industrial applications. Agricultural wastes are used as an inexpensive source for several utilizable products throughout the world. So, searching for cellulase enzymes from fungal strains capable of utilizing agricultural wastes to increase productivity, reduce costs and overcome waste accumulation in the environment is very important to evaluate its potency as a bio-additive to detergent agents. RESULTS: In the current study, the previously identified fungal strain Aspergillus terreus MN901491 was screened and selected for cellulase production. Medium parameters were optimized using one-factor-at-a-time (OFAT) and multi-factorial (Plackett-Burman and Box-Behnken) design methods. OFAT showed the ability of the fungal strain to utilize agricultural wastes (corn cob and rice straw) as a substrate. Also, yeast extract was the best nitrogen source for enhancing cellulase productivity. The most significant variables were determined by Plackett-Burman Design (PBD) and their concentrations were optimized by Response Surface Methodology (RSM) using Box-Behnken Design (BBD). Among eleven independent variables screened by PBD, malt extract, (NH4)2SO4, and KCl were the most significant ones followed by rice straw which affected cellulase production positively. The ANOVA results particularly the R2-value of PBD (0.9879) and BBD (0.9883) confirmed the model efficiency and provided a good interpretation of the experiments. PBD and BBD improved cellulase productivity by 6.1-fold greater than that obtained from OFAT. Medium optimization using OFAT and statistical models increased cellulase production from A. terreus MN901491 by 9.3-fold compared to the non-optimized medium. Moreover, the efficiency of cellulase activity on cotton fabrics as a bio-additive detergent was evaluated and estimated using whiteness and scanning electron microscope (SEM) that affirmed its potential effect and remarkable detergent ability to improve whiteness by 200% in comparison with non-washed fabric and by 190% in comparison with fabric washed by water. CONCLUSION: The presented work was stabilized as a multi-efficiency in which wastes were used to produce cellulase enzyme from the fungal strain, Aspergillus terreus MN901491 as a bio-additive to detergent applications that involved ecofriendly and green processes.

Covalent immobilization of ß-galactosidase using a novel carrier alginate/tea waste: statistical optimization of beads modification and reusability.

ß-galactosidase has been immobilized onto novel alginate/tea waste gel beads (Alg/TW) via covalent binding. Alg/TW beads were subjected to chemical modification through amination with polyethyleneimine (PEI) followed by activation with glutaraldehyde (GA). Chemical modification parameters including PEI concentration, PEI pH, and GA concentration were statistically optimized using Response Surface methodology (RSM) based on Box-Behnken Design (BBD). Analysis of variance (ANOVA) results confirmed the great significance of the model that had F value of 37.26 and P value < 0.05. Furthermore, the R2 value (0.9882), Adjusted R2 value (0.9617), and predicted R2 value (0.8130) referred to the high correlation between predicted and experimental values, demonstrating the fitness of the model. In addition, the coefficient of variation (CV) value was 2.90 that pointed to the accuracy of the experiments. The highest immobilization yield (IY) of ß-galactosidase (75.1%) was given under optimized conditions of PEI concentration (4%), PEI pH (9.5), and GA concentration (2.5%). Alg/TW beads were characterized by FT-IR, TGA, and SEM techniques at each step of immobilization process. Moreover, the immobilized ß-galactosidase revealed a very good reusability as it could be reused for 15 and 20 consecutive cycles keeping 99.7 and 72.1% of its initial activity, respectively. In conclusion, the environmental waste (tea waste) can be used in modern technological industries such as the food and pharmaceutical industry.

Antibacterial Evaluation of Gallic Acid and its Derivatives against a Panel of Multi-drug Resistant Bacteria.

BACKGROUND: Infectious diseases are the second leading cause of deaths worldwide. Pathogenic bacteria have been developing tremendous resistance against antibiotics which has placed an additional burden on healthcare systems. Gallic acid belongs to a naturally occurring phenolic class of compounds and is known to possess a wide spectrum of antimicrobial activities. AIMS & OBJECTIVES: In this study, we synthesized thirteen derivatives of gallic acid and evaluated their antibacterial potential against seven multi-drug resistant bacteria, as well as cytotoxic effects against human embryonic kidney cell line in vitro. Methods: 13 compounds were successfully synthesized with moderate to good yield and evaluated. Synthesized derivatives were characterized by using nuclear magnetic resonance spectroscopy, mass spectrometry, and Fourier transformation infrared spectroscopy. Antibacterial activity was determined using microdilution while cytotoxicyt was assessed using MTT assay. RESULTS: The results of antibacterial assay showed that seven out of thirteen compounds exhibited antibacterial effects with compound 6 and 13 being most potent against Staphylococcus aureus (MIC 56 µg/mL) and Salmonella enterica (MIC 475 µg/mL) respectively. On the other hand, most of these compounds showed lower cytotoxicity against human embryonic kidney cells (HEK 293), with IC50 values ranging from over 700 µg/mL. CONCLUSION: Notably, compound 13 was found to be non-toxic at concentrations as high as 5000 µg/mL. These findings suggest that the present synthetic derivatives of gallic acid hold potential for further studies in the development of potent antibacterial agents.

The role of landscape management practices to address natural resource degradation and human vulnerability in Awash River basin, Ethiopia.

Landscape management practices (LMP) support addressing the vulnerability of small-scale producers (SSPs) through providing a means of sustaining and strengthening community livelihoods and building their resilience and the environment. However, addressing the vulnerability of SSPs through the implementation of LMP requires meaningful community engagement and assessing the benefits and costs from the perspective of local communities. This study was conducted in two watersheds, Maybar-Felana and Gelana, in the Awash River basin, Ethiopia. The study assessed the links between natural resource degradation and the vulnerability of SSPs, local communities' opinion on the benefits and costs of LMP and the implications of implementing LMP for addressing vulnerability. It gathered and analyzed data through key informant interviews (KII), focus group discussions (FGDs) and GIS and remote sensing techniques. Diverse LMP such as afforestation/reforestation, exclosures, terrace and bunds and crop- and soil-based soil amendments were adopted in the studied watersheds. These practices contributed to the improvement of natural resources such as forests and the services they provide. Over the last 21 years (2000-2021), forest cover increased by 11.5 and 42.5% in Maybar-Felana and Gelana watersheds, respectively, while shrublands increased by 41.1% in Maybar-Felana. In line with this, the SSPs identified multiple benefits of LMP including the restoration of degraded vegetation, reducing runoff and soil loss, improving access to water for multiple uses and increasing agricultural productivity. The adopted LMP contributed to reducing livelihood vulnerability through reducing incidents of weather extremes such as flood and drought, improving food and water security, enhancing resource availability, and building livelihood assets. The SSPs also identified multiple economic and social costs of LMP, suggesting that addressing the economic and social costs through balancing short-term economic losses with long-term environmental benefits of interventions is crucial to sustaining the LMP and the benefits they provide.

Identification of potent anti-Candida metabolites produced by the soft coral associated Streptomyces sp. HC14 using chemoinformatics.

Candida albicans is the most common pathogen responsible for both spontaneous and recurrent candidiasis. The available treatment of Candida infections has several adverse effects, and the development of new drugs is critical. The current study looked at the synthesis of anti-Candida metabolites by Streptomyces sp. HC14 recovered from a soft coral. Using the Plackett Burman design, the medium composition was formulated to maximize production. Using GC-MS, the compounds have been identified, and a cheminformatics approach has been used to identify the potential source of activity. The compounds that showed high potential for activity were identified as pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(phenylmethyl)-3 and di-n-octyl based on their docking score against the cytochrome monooxygenase (CYP51) enzyme in Candida albicans. As a result of their discovery, fewer molecules need to be chemically synthesized, and fermentation optimization maximizes their synthesis, providing a strong foundation for the development of novel anti-Candida albicans agents.

Protease immobilization on a novel activated carrier alginate/dextrose beads: Improved stability and catalytic activity via covalent binding.

Protease from Bacillus thuringiensis strain-MA8 was successfully immobilized onto activated Alginate/dextrose (Alg/dex) beads as a new carrier with immobilization yield 77.6 %. The carrier was characterized using Scanning electron microscopy and Fourier transforms infrared spectrophotometer at every step of the immobilization process. Immobilized protease showed an increase of 10 °C in the optimum temperature compared to the free enzyme. However, the optimum pH for both the free and the Alg/dex/protease was found to be 8. The lower activation energy and deactivation rate constant and the higher half-life time and D-value confirm that the new Alg/dex carrier is suitable for promoting enzyme stability. The raise in thermal stability is also shown by the increased deactivation energy of the Alg/dex/protease compared to its free form by 1.47-fold. Likewise, the enzyme immobilization enhancement of Alg/dex/protease was accompanied by a marked increase in enthalpy and Gibbs free energy. The negative entropy for both free and Alg/dex/protease indicates that the enzyme is more stable in thermal deactivation. The Km and Vmax for the Alg/dex/protease were 2.05 and 1.22-times greater than the free form. Furthermore, Alg/dex/protease displayed good reusability as it retained 92.7 and 52.4 % of its activity after 8 and 12 hydrolysis cycles.

Statistical improvement of protease production from a new isolate Bacillus thuringiensis strain-MA8 and its application in the production of enzyme-modified cheese.

Enzyme-modified cheese (EMC) is a concentrated cheese flavor that is produced enzymatically from dairy substrates to provide an intense source of cheese flavor with broad applications. In this study, EMC was produced by enzymatic biotransformation from a new bacterial isolate described and molecularly identified as Bacillus thuringiensis strain-MA8. Optimization of protease production conditions using one-variable-at-a-time followed by multi-factorial (Plackett-Burman and Box-Behnken) designs increased production by 7-fold. Protease was used at different concentrations (300 and 900 U/100 g curd) as a cost-effective source of concentrated cheese flavor in the EMC preparation. Sensorial evaluation of EMC revealed that the overall acceptability, flavor, and texture were improved from the 2nd day compared to the control, and then decreased on the 4th day without any apparent bitterness. The chemical characteristics of EMC showed that the addition of protease extracts increased the total volatile fatty acids, water-soluble nitrogen, and acidity of EMC significantly (p≤0.05) compared to the control. The amino acids profile revealed that EMC1 which was treated with (300 U/100 g curd) protease had the highest essential amino acids (EAA) and EAA/total amino acids ratio. Nutritional parameters including protein efficiency ratio, biological value, and chemical score of EMC were higher than control based on Val, Met + Cys, Ile, Leu, and Phe + Tyr amino acids. Also, Scanning Electron Microscopy showed significant changes in EMC compared to the control. In conclusion, the addition of (300 U/100g curd) of protease revealed good EMC characteristics without any apparent defect.

Improving the reproductive, immunity and health status of rabbit does using honey bee venom.

The aim of the present study was to investigate the long-term effect of using bee venom (BV) on the reproductive performance, immune, and health status of rabbit does and its effect on their litters. Sixty mature does, from Spanish V-line rabbit stock, were randomly assigned to four homogeneous groups with 15 does each. The 1st , 2nd and 3rd groups were injected twice weekly under the neck skin with 0.1 ml solution contains 0.1, 0.2 and 0.3 mg BV/rabbit respectively. The 4th group served as a control group. From the results, litter size at birth, litter weight and survival rate at weaning age as well as milk yield were significantly (p ≤ 0.05) increased in BV groups than in the control group. Serum estradiol 17-ß (E2) was significantly (p ≤ 0.05) higher (15%) in the rabbit does treated with BV compared to the control group. The treated does with BV at any study doses showed a gradual and significant (p ≤ 0.05) decrease (12%) in serum progesterone levels (P4) compared to the control. They also showed a significant (p≤0.05) increase in conception (17%) and fertility rates (10%) compared to the control does. Treatment of rabbit does with BV caused a gradual and significant (p ≤ 0.05) reduction in both aspartate aminotransferase (AST) (16%) and alanine aminotransferase (ALT) (37%) liver enzyme activities. Additionally, results have shown that BV resulted in a gradual and significant (p ≤ 0.05) increase in total antioxidant capacity (TAC), antioxidative enzymes such as glutathione S-transferase (GST) and glutathione peroxidase (GPx), serum IgG, IgM and IgA levels with significant (p ≤ 0.05) decrease in malondialdehyde (MDA) and thiobarbituric acid reactive substances (TBARS) in BV groups compared to the control group. Results suggest that BV can be used in rabbit farming as an effective and safe alternative to artificial chemical drugs (sexual-stimulants) to improve certain reproductive traits, immune response and health.

Corrigendum to "Catalytic, kinetic and thermal properties of free andimmobilized Bacillus subtilis -MK1 α-amylase on Chitosan-magnetic nanoparticles" [Biotechnol. Rep. 26 (2020) e00443].

[This corrects the article DOI: 10.1016/j.btre.2020.e00443.].

Effect of bee venom on reproductive performance and immune response of male rabbits.

The current study aimed to investigate the effects of injecting bucks with different doses of bee venom (BV) on reproductive performance and immune response during the summer season. Forty-eight male V-line rabbits were randomly distributed among four homogeneous groups (12 bucks each). Three groups were injected BV under the neck skin with 0.1 (G1), 0.2 (G2) and 0.3 (G3) mg/rabbit twice weekly over 20 wks of treatment period. The 4th group (G0) was not injected BV and served as a control group. Buck groups that were treated with BV showed significantly (p≤0.05) shorter reaction time (increased libido) compared to the control, and this effect was BV dose-dependent manner. Viable sperm and concentration, total sperm output, live sperm, and fertility percentage were significantly (p≤0.05) higher in BV groups than in the control group. Additionally, testosterone concentration, and some other blood biochemical constituents (total protein, albumin, and glucose) were significantly (p≤0.05) higher in BV groups than those in the control group. The BV doses resulted in a significant (p≤0.05) increase of antioxidant indices (TAC, GST, and GSH) compared to the control group. Furthermore, the Immunoglobulin A (IgA) and immunoglobulin M (IgM) were significantly (p≤0.05) higher in BV groupscompared to the control group. From these results, we concluded that BV had significant positive impacts on some semen quality traits, sexual behavior, blood biochemical parameters, blood antioxidant content, lipid peroxidation biomarkers, and immune response in V-line bucks.

Catalytic, kinetic and thermal properties of free andimmobilized Bacillus subtilis -MK1 α-amylase on Chitosan-magnetic nanoparticles.

Bacillus subtilis strain-MK1 α-amylase was successfully immobilized on Chitosan-magnetic nanoparticles (Ch-MNP) that had been modified with polyethyleneimine (PEI) and glutaraldehyde (GA). Optimization of Ch-MNP/PEI/GA beads modification by Central Composite design enhanced the immobilization yield (IY %) by 1.5-fold. Ch-MNP/PEI/GA was characterized before and after modification and immobilization by FTIR and SEM. Ch-MNP/PEI/GA/Enzyme showed the same pH optima of free enzyme, while an elevation 10 °C in temperature optima was observed after its immobilization. Ch-MNP/PEI/GA/Enzyme displayed higher Km and Vmax values (2.1 and 1.2-fold) and lower Vmax/Km ratio (1.7-fold), respectively than the free enzyme. Compared to the free enzyme, Ch-MNP/PEI/GA/Enzyme exhibited lower activation energy, lower deactivation constant rate, higher D-values, higher half-life, and higher energy for denaturation. Immobilization of α-amylase increased enthalpy (4.2-fold), free energy (1.1-fold) and decreased entropy (4.6-fold) of thermal inactivation. A significant increase in pH stability of Ch-MNP/PEI/GA/Enzyme was observed especially at alkaline pH values. In addition, Ch-MNP/PEI/GA/Enzyme preserved 83.2 % of its initial activity after 15 consecutive cycles. When storing Ch-MNP/PEI/GA/Enzyme at 4 °C the residual activity was 100 and 86 %, respectively after 21 and 40 days. Finally, immobilization process improved the catalytic properties and stabilities, thus raising the suitability for industrial processes with lower cost and time.

Effect of carbon dioxide during the early stage of duck egg incubation on hatching characteristics and duckling performance.

Gaseous exchange is very important for embryonic development during the incubation of bird eggs. The present study was carried out to investigate the effect of carbon dioxide (CO2) pumping during the first 10 days of Pekin duck egg incubation on hatching characteristics, embryonic growth, hormonal concentrations and the post-hatch weight of the ducks. Two different ventilation conditions were used in this study. In one condition (V), the incubator was ventilated standard. In the other condition (NV), CO2 was gradually pumped into a non-ventilated incubator to reach and maintain a 1% concentration through the first 10 days of incubation, after which the incubator was ventilated for the rest of the incubation period. The CO2 percentage in the V incubator was constant at approximately 0.18%; by contrast, in the NV incubator, the concentration was increased gradually from 0.17% to 1%, where it was maintained through the 10th day of incubation. Throughout the incubation period, the egg weight loss and the embryonic mortality percentages were significantly lower in NV than in V. The hatchability of fertile eggs and the weights of ducklings at hatch were significantly higher in NV than in V. The embryonic T3, T4 and corticosterone levels in NV were significantly higher than those in V. Additionally, the haematological parameters (haemoglobin, packed cell volume and red blood cells) of the embryos were significantly higher in NV than in V. Furthermore, a total of 1198 ducklings for the V and NV conditions, all one day old, were individually weighed and then transferred into randomly assigned floor pens with 3 replicates for each ventilation condition. The feed conversion ratio of the ducks was significantly lower in NV than in V, while the reverse was true of body weight at different ages. Therefore, we can conclude that the NV condition with circulation of CO2 for the first 10 days of incubation is preferable to the V condition.

Outpatient balloon vaginoplasty for treatment of vaginal aplasia: retropubic balloon vaginoplasty was performed as an office procedure under local anesthesia.

We evaluated the feasibility and operative and postoperative outcomes of office balloon vaginoplasty. Office balloon vaginoplasty was performed successfully for a case with vaginal aplasia. She was currently married and presenting with minimal penetration and dyspareunia. We successfully performed the procedure through retro-pubic space under local anesthesia within 25 min total operative time and it was well tolerated by the patient. No operative complications were reported. The resulting neovagina was cosmetically attractive and 10 cm in depth. Sexual intercourse was started on the day of catheter removal. Penetration and satisfaction scores increased up to 90 points for both partners.

Ferulsinaic acid, a sesquiterpene coumarin with a rare carbon skeleton from Ferula species.

Fractionation of methylene chloride extracts of the resin of Ferula vesceritensis and F. sinaica afforded three sesquiterpene coumarins and a glucose derivative. One of them was a sesquiterpene with a rare carbon skeleton. The structures of these compounds were determined by extensive NMR studies, including DEPT, COSY, NOE, HMQC, and HMBC.