Enhancement of clavulanic acid production by Streptomyces sp MU-NRC77 via mutation and medium optimization

Purpose: To enhance clavulanic acid production using UV-mutagenesis on Streptomyces sp. NRC77.Methods: UV-mutagenesis was used to study the effect of Streptomyces sp. NRC77 on CA production. Phenotypic and genotypic identification methods of the promising mutant strain were characterized. Optimization of the fermentation medium and culture conditions were investigatedResults: Out of the screened mutants, 120A3 mutant isolate was selected as promising. The phenotypic properties of 120A3 mutant showed culture characteristics similar to those of Streptomyces species. Phylogenetic analysis of 16S rRNA gene sequence indicate that this strain has similarity (99 %) to Streptomyces sp.T2-7; therefore it was suggested as Streptomyces sp. MU-NRC77 and has Gen Bank accession no. KT953342.Conclusion: Improvement of CA yield by 48 % was obtained from fermentation medium and culture condition optimization. Further optimization by addition of H2O2 and activated charcoal to the production medium increased CA yield to 646.12 and 682.94 mg/L respectively, i.e., 83 % more than that obtained prior to addition.Keywords: Clavulanic acid, Medium optimization, Phenotypic and Genotypic identification, Streptomyces sp. MU-NRC77, UV-Mutagenesi

Modifying the design of pond production systems can improve the health and welfare of farmed Nile tilapia, Oreochromis niloticus

Infected fish have been known to recover from mild illness when they are able to locate to warmer water. This study aimed to replicate this 'behavioural fever' effect in an aquaculture setting by artificially heating a section of a fish pond (thereby introducing a thermal gradient) and effectively modifying pond design. This was achieved through the construction of a 'greenhouse' type structure above a section of the pond. Over the length of the production cycle at three typical Nile tilapia (Oreochromis niloticus) farms, the study collected data on water quality and fish growth and at the end of the cycle, blood samples were taken and total production was recorded. At each farm, fish were divided into two identical ponds, one with a greenhouse covering 3% of the pond surface area and one without. Results showed that greenhouse was effective in warming the surface of the water immediately below it. Oxygen levels were also higher under the greenhouse than outside of it and higher than in the control pond. Fish reared in the greenhouse ponds tended to be larger than the control ponds and had improved physiological and immune status (i.e., better liver and kidney function, higher antioxidant activity and lysozyme count; p < 0.05). The results of this study suggest that low-cost interventions that introduce thermal gradients in aquaculture systems may hold promise for improving health and welfare status of farmed fish in developing countries

Modifying the design of pond production systems can improve the health and welfare of farmed Nile tilapia, Oreochromis niloticus

Infected fish have been known to recover from mild illness when they are able to locate to warmer water. This study aimed to replicate this 'behavioural fever' effect in an aquaculture setting by artificially heating a section of a fish pond (thereby introducing a thermal gradient) and effectively modifying pond design. This was achieved through the construction of a 'greenhouse' type structure above a section of the pond. Over the length of the production cycle at three typical Nile tilapia (Oreochromis niloticus) farms, the study collected data on water quality and fish growth and at the end of the cycle, blood samples were taken and total production was recorded. At each farm, fish were divided into two identical ponds, one with a greenhouse covering 3% of the pond surface area and one without. Results showed that greenhouse was effective in warming the surface of the water immediately below it. Oxygen levels were also higher under the greenhouse than outside of it and higher than in the control pond. Fish reared in the greenhouse ponds tended to be larger than the control ponds and had improved physiological and immune status (i.e., better liver and kidney function, higher antioxidant activity and lysozyme count; p < 0.05). The results of this study suggest that low-cost interventions that introduce thermal gradients in aquaculture systems may hold promise for improving health and welfare status of farmed fish in developing countries.Article Information Received 26 September 2022 Revised 05 November 2022 Accepted 28 November 2022; O n l i n e F i r s t A r t i c l

The Effects of Wheat Flour and Barley Flour on the Quality and Properties of Biscuits Colored with Synthetic and Natural Colorants

Biscuit is one of the most popular processed ready to eat snacks that have possesses several attractive features including wide consumer base, relatively less expensive, more convenient with long shelf-life and have ability to serve as vehicles for important nutrients. It’s usually available in different sizes, tastes and shapes.  It can also be enriched or fortified with other ingredients in order to meet specific nutritional or therapeutic needs of consumers. In recent years, numerous studies have shown the potential of utilizing natural plant such as green leafy vegetable in cookies and biscuit production; the potential of amaranthus leaves used as a source of iron and β-carotene inpakora, vada, namakpal, kurmure, biscuit and cake. The present research was carried out to throw the light on the effect of wheat flour and barley flour with natural and synthetic colors to improve chemical composition, the nutritional value and quality characteristics of producing biscuits. The results indicate that that treatment 100% barley flour 72% with turmeric and cocoa as a natural colors recorded a significant increase in the content of protein, ash, minerals such as iron, zinc, potassium, magnesium and some physical analysis like weight, volume, density as well as sensory evaluation. As compared with wheat flour 72% and synthetic color tartrazine ADI 7.5 mg/kg.bw and chocolate brown ADI 0.15 mg/kg.bw. In conclusion, results showed that barley flour improved nutritional quality, physical characteristics and sensory characteristics of produce biscuit with mixture natural and natural color

Biochemical Changes in Fatty Acids, Hydrocarbons and Sterols as well as Total Lipids of Albino Rats Ingested some Synthetic Colourants and Flavourants as Food Additives

The present study was carried out to investigate the effect of ingested synthetic food colourants or flavourants on total lipids content feces. The feces of rats ingested synthetic food colourants or flavourants has more lipids than that of control feces. In treated rat feces some fatty acids were disappeared (C19) or appeared (C14) while the other were increased (C15, C20, C18:2) or decreased (C6) when compared with the control feces. Most of unsaturated fatty acids (un SFA) might bind with the synthetic food colourants or flavourants and secreted in feces, but less consumed than saturated fatty acids (SFA) which, observed on hydrocarbon components, but sterols including cholesterol were more secreted by synthetic food additives treatments. Generally, the increase of total lipids and lipid fractions in feces such as unsaturated fatty acids (un SFA), total sterols (TS) especially cholesterol maybe due to their abilities to bind with food additives and form complex which secreted in feces

Biochemical Changes in Fatty Acids, Hydrocarbons and Sterols as well as Total Lipids of Albino Rats Ingested some Synthetic Colourants and Flavourants as Food Additives

The present study was carried out to investigate the effect of ingested synthetic food colourants or flavourants on total lipids content feces. The feces of rats ingested synthetic food colourants or flavourants has more lipids than that of control feces. In treated rat feces some fatty acids were disappeared (C19) or appeared (C14) while the other were increased (C15, C20, C18:2) or decreased (C6) when compared with the control feces. Most of unsaturated fatty acids (un SFA) might bind with the synthetic food colourants or flavourants and secreted in feces, but less consumed than saturated fatty acids (SFA) which, observed on hydrocarbon components, but sterols including cholesterol were more secreted by synthetic food additives treatments. Generally, the increase of total lipids and lipid fractions in feces such as unsaturated fatty acids (un SFA), total sterols (TS) especially cholesterol maybe due to their abilities to bind with food additives and form complex which secreted in feces

Chromosomal aberrations and nucleic acids systems affected by some Egyptian medicinal plants used in treating female pregnant diabetic rats

ABSTRACT The influences of medicinal plants Juniperus Phoenicea (Araar), Hyphaene thebaica (Doum), Anastatica hierochuntica (Kafta) and Cleome droserifolia (Sammo) as antidiabetic agents were investigated using female pregnant albino rats. Female rats were injected with 60 mg/kg b.w. alloxan to induce diabetes. Diabetic rats treated orally with the methanol extracts of tested plants till the 19 day of gestation. The present studies include the frequencies of chromosomal aberrations and nucleic acid system of liver in the female pregnant rats and their embryos. The results showed that injection of alloxan caused highly significant increase in chromosomal aberrations as well as in blood glucose levels as a result of diabetes in pregnant females. It also caused a high incidence of chromosomal deviation in embryos and decreased the liver soluble protein contents of female rats and their embryos. These effects in alloxanized animals were treated and improved by ingestion of the methanol extracts of the tested plants (Araar, Doum, Kafta and Somma) in which under their treatments, the inceased level of blood glucose of diabetic rats was deceased. Ingestion with the plants methanolic extracts improved and normalized the effects of diabetes in nucleic acids values of liver tissues. These were accompanied with nucleases (RNAase and DNAase) activities. The inhibited activities of both DNA ase and RNA ase of pregnant rats and their embryos were stimulated and readjusted around the normal values. Also administration of the plants methanol extracts decreased the percentage of chromosomal aberrations in the female rats and embryos. It is concluded that there are some biochemical dynamics which might occur in the metabolism of glucose, nucleic acids and proteins in order to prevent or to reduce the oxidative stress of diabetes by flavonoides treatment

Phytochemical screening and in-vitro biological properties of unprocessed and household processed fenugreek (Trigonella foenum-graecum Linn.) seeds and leaves

Abstract The impact of household processes on fenugreek leaves and seeds has been analyzed for total phenolic (TP) and total flavonoid content (TF), and in-vitro biological activities such as antioxidant, antimicrobial, and anti-inflammatory properties. Processes included air-drying for leaves and germinating, soaking, and boiling for seeds. Air-dried fenugreek leaves (ADFL) had high TP (15.27 mg GAE g−1 D.W.) and TF (7.71 mg QE g−1 D.W.) (milligram quercetin equivalents per gram dry weight). The TP contents of unprocessed, germinated, soaked, and boiled seeds were 6.54, 5.60, 4.59, and 3.84 mg gallic acid equivalents per gram of dry weight (mg GAE g−1 D.W.), respectively. The TF contents in unprocessed fenugreek seeds, germinated fenugreek seeds, soaked fenugreek seeds, and boiled fenugreek seeds (BFS) were 4.23, 2.11, 2.10, and 2.33 mg QE g−1 D.W., respectively. Sixteen phenolic and nineteen flavonoid compounds has been identified using high-performance liquid chromatography. Antioxidant activity using 2,2-diphenyl-1-picrylhydrazil (DPPH·), 2,2-azinobis (3-ethylbenothiazoline-6-sulfonic acid (ABTS+·), and ferric reducing antioxidant power (FRAP·) assays indicated that ADFL had the highest activity. Antimicrobial activity has been evaluated against each of the eight pathogenic bacterial and fungal strains. ADFL showed the strongest activity with minimum inhibitory concentrations values ranging from 0.03 to 1.06 and 0.04 to 1.18 mg ml·1 against bacterial and fungal strains, respectively. Anti-inflammatory activity was evaluated in-vitro against RAW 264.7 macrophage cells using the nitric oxide (NO) assay. Results revealed that ADFL had the highest cytotoxicity and anti-inflammatory activity according to the NO assay. Household processes significantly reduced the in-vitro biological properties of processed seeds

Cell Growth Inhibition, DNA Fragmentation and Apoptosis-Inducing Properties of Household-Processed Leaves and Seeds of Fenugreek (<i>Trigonella Foenum-Graecum</i> Linn.) against HepG2, HCT-116, and MCF-7 Cancerous Cell Lines

Household processing of fenugreek seeds and leaves, including soaking, germination, and boiling of the seeds, and air-drying of the leaves, has improved the levels of human consumption of the bitter seeds and increased the shelf life of fresh leaves, respectively. The potential anticancer activity of either unprocessed or processed fenugreek seeds or leaves and the relative expression of pro-apoptotic and anti-apoptotic genes of the studied cancerous cell lines exposed to IC50 crude extracts was investigated to observe the apoptotic-inducing property of this plant as an anticancer agent. The protein expression of IKK-α and IKK-β, as inhibitors of NF-KB which exhibit a critical function in the regulation of genes involved in chronic inflammatory disorders, were studied in the tested cancerous cell lines. In this study, the anticancer activity of household-processed fenugreek leaves and seeds against HepG2, HCT-116, MCF-7, and VERO cell lines was measured using an MTT assay. DNA fragmentation of both HepG2 and MCF-7 was investigated by using gel electrophoresis. RT-PCR was used to evaluate the relative expression of each p53, caspase-3, Bax, and Bcl-2 genes, whereas ELISA assay determined the expression of caspase-3, TNF-α, and 8-OHDG genes. Western blotting analyzed the protein-expressing levels of IKK-α and IKK-β proteins in each studied cell line. Data showed that at 500 µg mL−1, ADFL had the highest cytotoxicity against the HepG2 and HCT-116 cell lines. Although, each UFS and GFS sample had a more inhibitory effect on MCF-7 cells than ADFL. Gel electrophoresis demonstrated that the IC50 of each ADFL, UFS, and GFS sample induced DNA fragmentation in HepG2 and MCF-7, contrary to untreated cell lines. Gene expression using RT-PCR showed that IC50 doses of each sample induced apoptosis through the up-regulation of the p53, caspase-3, and Bax genes and the down-regulation of the Bcl-2 gene in each studied cell line. The relative expression of TNF-α, 8-OHDG, and caspase-3 genes of each HepG2 and MCF-7 cell line using ELISA assays demonstrated that ADFL, UFS, and GFS samples reduced the expression of TNF-α and 8-OHDG genes but increased the expression of the caspase-3 gene. Protein-expressing levels of IKK-α and IKK-β proteins in each studied cell line, determined using Western blotting, indicated that household treatments decreased IKK-α expression compared to the UFS sample. Moreover, the ADFL and SFS samples had the most activity in the IKK-β expression levels. Among all studied samples, air-dried fenugreek leaves and unprocessed and germinated fenugreek seeds had the most anti-proliferative and apoptotic-inducing properties against human HepG2, MCF-7, and HCT-116 cell lines, as compared to the VERO cell line. So, these crude extracts can be used in the future for developing new effective natural drugs for the treatment of hepatocellular, breast, and colon carcinomas

Cell Growth Inhibition, DNA Fragmentation and Apoptosis-Inducing Properties of Household-Processed Leaves and Seeds of Fenugreek (Trigonella Foenum-Graecum Linn.) against HepG2, HCT-116, and MCF-7 Cancerous Cell Lines

Household processing of fenugreek seeds and leaves, including soaking, germination, and boiling of the seeds, and air-drying of the leaves, has improved the levels of human consumption of the bitter seeds and increased the shelf life of fresh leaves, respectively. The potential anticancer activity of either unprocessed or processed fenugreek seeds or leaves and the relative expression of pro-apoptotic and anti-apoptotic genes of the studied cancerous cell lines exposed to IC50 crude extracts was investigated to observe the apoptotic-inducing property of this plant as an anticancer agent. The protein expression of IKK-&alpha; and IKK-&beta;, as inhibitors of NF-KB which exhibit a critical function in the regulation of genes involved in chronic inflammatory disorders, were studied in the tested cancerous cell lines. In this study, the anticancer activity of household-processed fenugreek leaves and seeds against HepG2, HCT-116, MCF-7, and VERO cell lines was measured using an MTT assay. DNA fragmentation of both HepG2 and MCF-7 was investigated by using gel electrophoresis. RT-PCR was used to evaluate the relative expression of each p53, caspase-3, Bax, and Bcl-2 genes, whereas ELISA assay determined the expression of caspase-3, TNF-&alpha;, and 8-OHDG genes. Western blotting analyzed the protein-expressing levels of IKK-&alpha; and IKK-&beta; proteins in each studied cell line. Data showed that at 500 &micro;g mL&minus;1, ADFL had the highest cytotoxicity against the HepG2 and HCT-116 cell lines. Although, each UFS and GFS sample had a more inhibitory effect on MCF-7 cells than ADFL. Gel electrophoresis demonstrated that the IC50 of each ADFL, UFS, and GFS sample induced DNA fragmentation in HepG2 and MCF-7, contrary to untreated cell lines. Gene expression using RT-PCR showed that IC50 doses of each sample induced apoptosis through the up-regulation of the p53, caspase-3, and Bax genes and the down-regulation of the Bcl-2 gene in each studied cell line. The relative expression of TNF-&alpha;, 8-OHDG, and caspase-3 genes of each HepG2 and MCF-7 cell line using ELISA assays demonstrated that ADFL, UFS, and GFS samples reduced the expression of TNF-&alpha; and 8-OHDG genes but increased the expression of the caspase-3 gene. Protein-expressing levels of IKK-&alpha; and IKK-&beta; proteins in each studied cell line, determined using Western blotting, indicated that household treatments decreased IKK-&alpha; expression compared to the UFS sample. Moreover, the ADFL and SFS samples had the most activity in the IKK-&beta; expression levels. Among all studied samples, air-dried fenugreek leaves and unprocessed and germinated fenugreek seeds had the most anti-proliferative and apoptotic-inducing properties against human HepG2, MCF-7, and HCT-116 cell lines, as compared to the VERO cell line. So, these crude extracts can be used in the future for developing new effective natural drugs for the treatment of hepatocellular, breast, and colon carcinomas