Search results for: herb
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 99

Search results for: herb

39 Determination of Heavy Metal Levels in Carissa spinarum and Toddalia asiatica Used as Herbal Medicines in Kisii and Nyamira Counties Region, Kenya

Authors: Moses A. Guto Maobe, Leonard Gitu, Erastus Gatebe

Abstract:

The plants Carissa spinarum and Toddalia asiatica have historically been used as herbal medicines in Kisii and Nyamira Counties region, Kenya. But, there is limited study about heavy metal contents in their different plant parts. Such information is necessary for proper use of the two plant species as herbal medicines. So, precise determination of heavy metal contents in different part of these herbs is required for quality, efficacy and safety use in the treatment of ailments. The main aim of this study was to standardize the two herbs of interest. The objective of this study was to evaluate the levels of heavy metal contents in the root of Carissa spinarum and Toddalia asiatica. A wet digestion method with concentrated nitric-hydrochloric acid was used for the dissolution of each herb part prior to elemental analysis. Standard solutions of various concentrations of each pure metal of analytical grade arsenic (As), cadmium (Cd) and mercury (Hg) were prepared and used. The analysis of As, Cd and Hg in each of two herbs was conducted by atomic absorption spectroscopy (AAS) Shimadzu model No. 6200. Data obtained from root of Carissa spinarum indicated concentration (mgkg⁻¹) of Arsenic (As), Cadmium (Cd) and Mercury (Hg) were 0.87 x 10⁻³, 7.02 x 10⁻⁶ and 0.66 x 10⁻³ respectively. Results obtained from root of Toddalia asiatica showed concentration (mgkg⁻¹) of Arsenic (As), Cadmium (Cd) and Mercury (Hg) were 1.33 x 10⁻³, 7.32 x 10⁻⁶ and 1.13 x 10⁻³, respectively. The permissible limits set by WHO for As, Cd and Hg in herbs are (mgkg⁻¹) < 1 - 5, < 0.3 – 1 and < 0.1- 0.5 respectively. The concentrations of As, Cd, and Hg determined were relatively higher in the root of Toddalia asiatica than the root of Carissa spinarum. It was concluded that levels of heavy metal contents of As, Cd, and Hg in the root of Carissa spinarum and Toddalia asiatica were within permissible limits set by WHO/FAO.

Keywords: heavy metals, Carissa spinarum, Toddalia asiatica, wet digestion, pollutants, AAS

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38 Comparative Therapeutic Effect of Acalypha indica Linn. Extract and Gemfibrozil on High Fructose and Cholesterol Diet Induced Pancreas Steatosis in Sprague-Dawley Mice

Authors: Adrian Reynaldo Sudirman, Siti Farida, Aisyah Aminy Maulidina, Caren Andika Surbakti

Abstract:

Sedentary lifestyle and imbalance consumption pattern has made metabolic syndrome as the global time bomb phenomenon in the world. The increasing tendency of people in consuming high amount of fructose and cholesterol food has worsened this issue in the society. Pancreas steatosis become one of the most comorbid when early diagnosis and prompt treatment has not been applied on hyperglycemic and hyperlipidemic condition in metabolic syndrome patient. Gemfibrozil become the drug of choice to prevent this issue, yet the efficacy of this regiment was still questionable. Acalypha indica Linn. is the herb that has protective effect on hyperlipidemic and hyperglycemic condition. This study was aimed to compare therapeutic effect of gemfibrozil (G) and Acalypha indica Linn. (AI) on high fructose and cholesterol diet-induced pancreas steatosis in Sprague-Dawley mice. The post induction mice were divided into four groups: control, gemfibrozil, AI extract, and G+AI combination regiment. Each group received four weeks intervention. The result of statistical analysis using the One-Way ANOVA test and Tukey Post Hoc test showed significant decrease in pancreatic steatosis between the control group and administered Acalypha indica group (p = 0.004, 95% CI: 0.170-0.959) and the group administered with a combination of Gemfibrozil-Acalypha indica (p = 0.023, 95% CI: 0.537-0.813). The protective effect of Acalypha indica Linn. shows that this plant has the potential as therapeutic option in overcoming the condition of pancreas steatosis in metabolic syndrome.

Keywords: Acalypha Indica Linn., Cholesterol, Fructose, Gemfibrozil, Pancreas Steatosis

Procedia PDF Downloads 279
37 Effect of Cellulase Pretreatment for n-Hexane Extraction of Oil from Garden Cress Seeds

Authors: Boutemak Khalida, Dahmani Siham

Abstract:

Garden cress (Lepidium Sativum L.) belonging to the family Brassicaceae, is edible growing annual herb. Its various parts (roots, leaves and seeds) have been used to treat various human ailments. Its seed extracts have been screened for various biological activities like hypotensive, antimicrobial, bronchodilator, hypoglycaemic and antianemic. The aim of the present study is to optimize the process parameters (cellulase concentration and incubation time) of enzymatic pre-treatment of the garden cress seeds and to evaluate the effect of cellulase pre-treatment of the crushed seeds on the oil yield, physico-chemical properties and antibacterial activity and comparing to non-enzymatic method. The optimum parameters of cellulase pre-treatment were as follows: cellulase of 0,1% w/w and incubation time of 2h. After enzymatic pre-treatment, the oil was extracted by n-hexane for 1.5 h, the oil yield was 4,01% for cellulase pre-treatment as against 10,99% in the control sample. The decrease in yield might be caused a result of mucilage. Garden cress seeds are covered with a layer of mucilage which gels on contact with water. At the same time, the antibacterial activity was carried out using agar diffusion method against 4 food-borne pathogens (Escherichia coli, Salmonella typhi,Staphylococcus aureus, Bacillus subtilis). The results showed that bacterial strains are very sensitive to the oil with cellulase pre-treatment. Staphylococcus aureus is extremely sensitive with the largest zone of inhibition (40 mm), Escherichia coli and salmonella typhi had a very sensitive to the oil with a zone of inhibition (26 mm). Bacillus subtilizes is averagely sensitive which gave an inhibition of 16 mm. But it does not exhibit sensivity to the oil without enzymatic pre-treatment with a zone inhibition (< 8 mm). Enzymatic pre-treatment could be useful for antimicrobial activity of the oil, and hold a good potential for use in food and pharmaceutical industries.

Keywords: Lepidium sativum L., cellulase, enzymatic pretreatment, antibacterial activity.

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36 The Treatment Effect of Turmeric (Curcuma domestica Val.) Rhizome Extract to Reduce Serum Transaminase Level on Paracetamol Induced Liver Toxicity in Wistar White Male Rats (Rattus norvegicus)

Authors: David Tanujaya Kurniawan

Abstract:

Background: Liver injury caused by paracetamol is marked by increased serum transaminase levels. Turmeric is a local herb that is available in large quantities and inexpensive in contradiction to its substantial benefits, including its potency to increase glutathione production and regenerate hepatocyte into normal condition. Aim: The aim of this study was to analyze the potencial treatment effect of turmeric rhizome extract to reduce serum transaminase level on paracetamol induced liver toxicity in rats. Methods: This study was a laboratory experimental research with post-test only controlled group design. A group of 24 Wistar white male rats was induced with paracetamol 360 mg/kg body weight for 10 days. The group was then separated into four groups: the first and the second was treated with 100 mg/kg body weight and 150 mg/kg body weight of turmeric rhizome extract, subsequently, the third as positive control was given 27 mg/kg body weight of lesichol, while the fourth as negative control was given CMC-Na 1%. Each of this treatment was given for seven days. At the end of the study, the blood samples were taken to measure SGOT and SGPT levels. The one-way Anova test revealed significant difference in mean of SGPT level (p=0,001). The LSD test showed significant differences of SGPT levels in both treatment groups and negative control group. However, there was no sgnificant difference between positive control and both treatment groups. Conclusion: Curcuma domestica Val. rhizome extract could not reduce SGOT level, but it reduced SGPT level significantly.

Keywords: Curcuma domestica val., SGOT, SGPT, paracetamol, liver toxicity

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35 Performances of Ashwagandha (Withania somnifera Duanal) as Affected by Method of Planting and Source of Nutrients

Authors: Ewon Kaliyadasa, U. L. B. Jayasinghe, S. E. Peiris

Abstract:

Ashwagandha (Withania sominifera Duanal) is an important medicinal herb belongs to family Solanaceae. This plant has raised its popularity after discovering anti stress and sex stimulating properties that mainly due to the presence of biologically active alkaloid compounds. Therefore it is vital to adapt to a proper agro technological package that ensure optimum growth of ashwagandha to obtain the finest quality without degrading pharmacologically active constituents. Organic and inorganic fertilizer mixtures were combined with direct seeding and transplanting as four different treatments in this study. Tuber fresh and dry weights were recorded up to twelve months starting from two months after sowing (MAS) while shoot height, root length, number of leaves, shoot fresh and dry weights and root: shoot ratio up to 6MAS. Results revealed that growth of ashwagandha was not affected significantly by method of planting or type of fertilizer or its combinations during most of the harvests. However, tubers harvested at 6MAS recorded the highest dry tuber weight per plant in all four treatments compared to early harvests where two direct seeded treatments are the best. Chemical comparison of these two treatments, direct seeding coupled with organic and inorganic fertilizer shown that direct seeding with organic treatment recorded the highest values for alkaloid and withaferine A content with lower percentage of fiber. Further these values are in concurring with the values of commercially available tuber samples. Having considered all facts, 6MAS can be recommended as the best harvesting stage to obtain high quality tubers of ashwagandha under local conditions.

Keywords: alkaloids, direct seeding, dry tuber weight, inorganic fertilizer, organic fertilizer, transplanting, withaferine a

Procedia PDF Downloads 315
34 Adaptive Response of Plants to Environmental Stress: Natural Oil Seepage; The Living Laboratory in Tramutola, Basilicata Region

Authors: Maria Francesca Scannone, Martina Bochicchio

Abstract:

One of the major environmental problems today is hydrocarbon contamination. The promising sustainable technologies for the treatment of these contaminated sites involves the use of biological organisms. In Agri Valley (Basilicata Region) there is a living laboratory (natural oil seeps) where the selective pressure has enriched the environmental matrices with microorganisms, fungi and plant species able to use the hydrocarbons as a source of metabolic energy, to degrade or tolerate hydrocarbons. Observers visiting this area are fascinated by its unspoiled nature, and the condition of the ecosystem does not appear to has been damaged. The amazing resiliency observed in Tramutola site is of key importance to try to bring green remediation technologies, but no research has been done to identify high-performing native species. The aim of this research was to study how natural processes affect the fate of released oil or how individual species or communities of plants and animals are capable of dealing with the burden of otherwise toxic chemicals. The survey of vegetation was carried out, more than 60 species have been identified and divided into tree, shrub and herb layer. Plant data sheets have been completed only for the species that showed the most appropriate properties for phytoremediation. In general, members of the Salicales, Cyperales, Poales, Fagales, Cornales, Equisetales orders were the most commonly identified orders. They are pioneer plants with high adaptive capacity and vegetative propagation. The literature review has highlighted the existence of rhizosphere effect and a green liver model on selected plants. The study provides significant information on the environmental stress adaptation processes of many indigenous plants that are living and growing on a natural leak of crude oil and gas that migrates up through subsurface.

Keywords: green liver, hydrocarbon degradation, oil seeps, phytoremediation

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33 Potential Application of Thyme (Thymus vulgaris L.) Essential Oil as Antibacterial Drug in Aromatherapy

Authors: Ferhat Mohamed Amine, Boukhatem Mohamed Nadjib, Chemat Farid

Abstract:

The Lamiaceae family is widely spread in Algeria. Due to the application of Thymus species growing wild in Algeria as a culinary herb and in folk medicine, the purpose of the present work was to evaluate antimicrobial activities of their essential oils and relate them with their chemical composition, for further application in food and pharmaceutical industries as natural valuable products. The extraction of the Thymus vulgaris L. essential oil (TVEO) was obtained by steam distillation. Chemical composition of the TVEO was determined by Gas Chromatography. A total of thirteen compounds were identified. Carvacrol (83.8%) was the major component, followed by cymene (8.15%) and terpinene (4.96%). Antibacterial action of the TVEO against 23 clinically isolated bacterial strains was determined by using agar disc diffusion and vapour diffusion methods at different doses. By disc diffusion method, TVEO showed potent antimicrobial activity against gram-positive bacteria more than gram-negative strains and antibiotic discs. The Diameter of Inhibition Zone (DIZ) varied from 25 to 60 mm for S. aureus, B. subtilisand E. coli. However, the results obtained by both agar diffusion and vapour diffusion methods were different. Significantly higher antibacterial effect was observed in the vapour phase at lower doses. S. aureus and B. subtilis were the most susceptible strains to the oil vapour. Therefore, smaller doses of EO in the vapour phase can be inhibitory to pathogenic bacteria. There is growing evidence that TVEO in vapour phase are effective antiseptic systems and appears worthy to be considered for practical uses in the treatment of human infections oras air decontaminants in hospital. TVEO has considerable antibacterial activity deserving further investigation for clinical applications. Also whilst the mode of action remains mainly undetermined, this experimental approach will need to continue.

Keywords: antimicrobial drugs, carvacrol, disc diffusion, Thymus vulgaris, vapour diffusion

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32 Critical Assessment of Herbal Medicine Usage and Efficacy by Pharmacy Students

Authors: Anton V. Dolzhenko, Tahir Mehmood Khan

Abstract:

An ability to make an evidence-based decision is a critically important skill required for practicing pharmacists. The development of this skill is incorporated into the pharmacy curriculum. We aimed in our study to estimate perception of pharmacy students regarding herbal medicines and their ability to assess information on herbal medicines professionally. The current Monash University curriculum in Pharmacy does not provide comprehensive study material on herbal medicines and students should find their way to find information, assess its quality and make a professional decision. In the Pharmacy course, students are trained how to apply this process to conventional medicines. In our survey of 93 undergraduate students from year 1-4 of Pharmacy course at Monash University Malaysia, we found that students’ view on herbal medicines is sometimes associated with common beliefs, which affect students’ ability to make evidence-based conclusions regarding the therapeutic potential of herbal medicines. The use of herbal medicines is widespread and 95.7% of the participated students have prior experience of using them. In the scale 1 to 10, students rated the importance of acquiring herbal medicine knowledge for them as 8.1±1.6. More than half (54.9%) agreed that herbal medicines have the same clinical significance as conventional medicines in treating diseases. Even more, students agreed that healthcare settings should give equal importance to both conventional and herbal medicine use (80.6%) and that herbal medicines should comply with strict quality control procedures as conventional medicines (84.9%). The latter statement also indicates that students consider safety issues associated with the use of herbal medicines seriously. It was further confirmed by 94.6% of students saying that the safety and toxicity information on herbs and spices are important to pharmacists and 95.7% of students admitting that drug-herb interactions may affect therapeutic outcome. Only 36.5% of students consider herbal medicines as s safer alternative to conventional medicines. The students use information on herbal medicines from various sources and media. Most of the students (81.7%) obtain information on herbal medicines from the Internet and only 20.4% mentioned lectures/workshop/seminars as a source of such information. Therefore, we can conclude that students attained the skills on the critical assessment of therapeutic properties of conventional medicines have a potential to use their skills for evidence-based decisions regarding herbal medicines.

Keywords: evidence-based decision, pharmacy education, student perception, traditional medicines

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31 Antioxidant Activity and Total Phenolic Content within the Aerial Parts of Artemisia absinthium

Authors: Hallal Nouria, Kharoubi Omar

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Wormwood (Artemisia absinthium L.) is a medicinal and aromatic bitter herb, which has been used as a medicine from ancient times. It has traditionally been used as anthelmintic, choleretic, antiseptic, balsamic, depurative, digestive, diuretic, emmenagogue and in treating leukemia and sclerosis. The species was cited to be used externally as cataplasm of crushed leaves for snake and scorpion bites or decoction for wounds and sores applied locally as antiseptic and antifungal. Wormwood extract have high contents of total phenolic compounds and total flavonoids indicating that these compounds contribute to antiradical and antioxidative activity. Most of the degenerative diseases are caused by free radicals. Antioxidants are the agents responsible for scavenging free radicals. The aim of present study was to evaluate the phytochemical and in vitro antioxidant properties of Wormwood extract. DPPH assay and reducing power assay were the method adopted to study antioxidant potentials of extracts. Standard methods were used to screen preliminary phytochemistry and quantitative analysis of tannin, phenolics and flavanoids. Aqueous and alcoholic extracts were showed good antioxidant effect with IC50 ranges from 62 μg/ml for aqueous and 116μg/ml for alcoholic extracts. Phenolic compounds, tannins and flavonoids were the major phytochemicals present in both the extracts. Percentage of inhibition increased with the increased concentration of extracts. The aqueous and alcoholic extract yielded 20, 15& 3, 59 mg/g gallic acid equivalent phenolic content 2, 78 & 1,83 mg/g quercetin equivalent flavonoid and 2, 34 & 6, 40 g tannic acid equivalent tannins respectively. The aqueous and methanol extracts of the aerial parts showed a positive correlation between the total phenolic content and the antioxidant activity measured in the plant samples. The present study provides evidence that both extracts of Artemisia absinthium is a potential source of natural antioxidant.

Keywords: pharmaceutical industries, medicinal and aromatic plant, antioxidants, phenolic compounds, Artemisia absinthium

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30 Optimization of Sucrose Concentration, PH Level and Inoculum Size for Callus Proliferation and Anti-bacterial Potential of Stevia Rebaudiana Bertoni

Authors: Inayat Ur Rahman Arshad

Abstract:

Stevia rebaudiana B. is a shrubby perennial herb of Asteraceae family that possesses the unique ability of accumulative non caloric sweet Steviol Glycosides (SGs). The purpose of the study is to optimize sugar concentration, pH level and inoculum size for inducing the callus with optimum growth and efficient antibacterial potential. Three different experiments were conducted in which Callus explant from three-months-old already established callus of Stevia reabudiana of four different sizes were inoculated on Murashige and Skoog (MS) basal medium supplemented with five different sucrose concentration and pH adjusted at four different levels. Maximum callus induction 100, 87.5 and 85.33% was resulted in the medium supplemented with 30g/l sucrose, pH maintained at 5.5 and inoculated with 1.25g inoculum respectively. Similarly, the highest fresh weight 65.00, 75.50 and 50.53g/l were noted in medium fortified with 40g/l sucrose, inoculated 1.25g inoculum and 6.0 pH level respectively. However, the callus developed in medium containing 50g/l sucrose found highly antibacterial potent with 27.3 and 26.5mm inhibition zone against P. vulgaris and B. subtilize respectively. Similarly, the callus grown on medium inoculated with 1.00g inoculum resulted in maximum antibacterial potential against S. aureus and P. vulgaris with 25 and 23.72mm inhibition zones respectively. However, in the case of pH levels the medium maintained at 6.5pH showed maximum antibacterial activity against P. vulgaris, B.subtilis and E.coli with 27.9, 25 and 23.72mm respectively. The ethyl acetate extract of Stevia callus and leaves did not show antibacterial potential against Xanthomonas campestris and Clavebactor michiganensis. In the entire experiment the standard antibacterial agent Streptomycin showed the highest inhibition zones from the rest of the callus extract, however the pure DMSO (Dimethyl Sulfoxide) caused no inhibitory zone against any bacteria. From these findings it is concluded that among various levels sucrose at the rate of 40g L-1, pH 6.0 and inoculums 0.75g was found best for most of the growth and quality attributes including fresh weight, dry weight and antibacterial activities and therefore can be recommended for callus proliferation and antibacterial potential of Stevia rebaudiana

Keywords: Steviol Glycosides, Skoog, Murashige, Clavebactor michiganensis

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29 Optimization of Sucrose Concentration, pH Level and Inoculum Size for Callus Proliferation and Anti-Bacterial Potential of Stevia rebaudiana Bertoni

Authors: Inayat Ur Rahman Arshad

Abstract:

Background: Stevia rebaudiana B. is a shrubby perennial herb of Asteraceae family that possesses the unique ability of accumulative non-caloric sweet steviol glycosides (SGs). Purpose: The purpose of the study is to optimize sugar concentration, pH level, and inoculum size for inducing the callus with optimum growth and efficient antibacterial potential. Method: Three different experiments were conducted in which Callus explant from three-months-old already established callus of Stevia reabudiana of four different sizes was inoculated on Murashige and Skoog (MS) basal medium supplemented with five different sucrose concentration and pH adjusted at four different levels. Results: Maximum callus induction 100, 87.5, and 85.33% resulted in the medium supplemented with 30 g/l sucrose, pH maintained at 5.5, and inoculated with 1.25g inoculum, respectively. Similarly, the highest fresh weights 65.00, 75.50, and 50.53 g/l were noted in a medium fortified with 40 g/l sucrose, inoculated 1.25g inoculum, and 6.0 pH level, respectively. However, the callus developed in a medium containing 50 g/l sucrose was found to be highly antibacterial potent with 27.3 and 26.5 mm inhibition zone against P. vulgaris and B. subtilis, respectively. Similarly, the callus grown on a medium inoculated with 1.00 g inoculum resulted in maximum antibacterial potential against S. aureus and P. vulgaris with 25 and 23.72 mm inhibition zone, respectively. However, in the case of pH levels, the medium maintained at 6.5 pH showed maximum antibacterial activity against P. vulgaris, B.subtilis, and E.coli with 27.9, 25, and 23.72 mm, respectively. The ethyl acetate extract of Stevia callus and leaves did not show antibacterial potential against Xanthomonas campestris and Clavebactor michiganensis. In the entire experiment, the standard antibacterial agent Streptomycin showed the highest inhibition zones among the rest of the callus extract; however, the pure dimethyl sulfoxide (DMSO) caused no inhibitory zone against any bacteria. Conclusion: From these findings, it is concluded that among various levels, sucrose @ 40 g L⁻¹, pH 6.0, and inoculums at 0.75 g were found best for most of the growth and quality attributes, including fresh weight, dry weight, and antibacterial activities and therefore can be recommended for callus proliferation and antibacterial potential of Stevia rebaudiana.

Keywords: Stevia rebaudiana, Steviol Glycosides, callus, Xanthomonas campestris

Procedia PDF Downloads 56
28 Synergistic Anti-Proliferation Effect of PLK-1 Inhibitor and Livistona Chinensis Fruit Extracts on Lung Adenocarcinoma A549 Cells

Authors: Min-Chien Su, Tzu-Hsuan Hsu, Guan-Xuan Wu, Shyh-Ming Kuo

Abstract:

Lung cancer is one of the clinically challenging malignant diseases worldwide. For efficient therapeutics in cancer, combination therapy has developed to acquire a better outcome. PLK-1 was one of the major factors affecting cell mitosis in cancer cells, its inhibitor Bi6727 was proven effective in treating several different cancers namely oral cancer, colon cancer and lung cancer. Despite its low toxicity toward normal cells compared to traditional chemotherapy, it is still yet to be evaluated in detail. Livistona Chinensis (LC) is a Chinese herb that used as a traditional prescription to treat lung cancer. Due to the uncertainty of the efficacy of LC, we utilized a water extraction method to extract the Livistona Chinensis and then lyophilized into powder for further study. In this study we investigated the antiproliferation activities of Bi6727 and LC extracts (LCE) on A549 non-small lung cancer cells. The IC50 of Bi6727 and LCE on A549 are 60 nM and 0.8 mg/mL, respectively. The fluorescent staining images shown nucleolus damage in cells treated with Bi6727 and mitochondrial damage after treated with LCE. A549 cells treated with Bi6727 and LCE showed increased expression of Bax, Caspase-3 and Caspase-9 proteins from Western blot assay. LCE also inhibited A549 cells growth keeping cells at G2-M phase from cell cycle assay. Apoptosis assay results showed that LCE induced late apoptosis of A549 cells. JC-1 assay showed that the mitochondria damaged at the LCE concentration of 0.4 mg/mL. In our preliminary anti-proliferation test of combined LCE and Bi-6727 on A549 cells, we found a dramatically decrease in proliferation after treated with LCE first for 24-h and then Bi-6727 for extra 24-h. This was an important finding regarding synergistic anti-proliferation effect of these drugs, However, the usage, the application sequence of LCE and Bi-6727 on A549 cells and their related mechanisms still need to be evaluated. In summary, the drugs exerted anti-proliferation effect on A549 cells independently. We hopefully combine the usage of these two drugs will bring a different and potential outcome in treating lung cancer.

Keywords: anti-proliferation, A549, Livistona Chinensis fruit extracts, PLK-1 inhibitor

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27 Application of a Synthetic DNA Reference Material for Optimisation of DNA Extraction and Purification for Molecular Identification of Medicinal Plants

Authors: Mina Kalantarzadeh, Claire Lockie-Williams, Caroline Howard

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DNA barcoding is increasingly used for identification of medicinal plants worldwide. In the last decade, a large number of DNA barcodes have been generated, and their application in species identification explored. The success of DNA barcoding process relies on the accuracy of the results from polymerase chain reaction (PCR) amplification step which could be negatively affected due to a presence of inhibitors or degraded DNA in herbal samples. An established DNA reference material can be used to support molecular characterisation protocols and prove system suitability, for fast and accurate identification of plant species. The present study describes the use of a novel reference material, the trnH-psbA British Pharmacopoeia Nucleic Acid Reference Material (trnH-psbA BPNARM), which was produced to aid in the identification of Ocimum tenuiflorum L., a widely used herb. During DNA barcoding of O. tenuiflorum, PCR amplifications of isolated DNA produced inconsistent results, suggesting an issue with either the method or DNA quality of the tested samples. The trnH-psbA BPNARM was produced and tested to check for the issues caused during PCR amplification. It was added to the plant material as control DNA before extraction and was co-extracted and amplified by PCR. PCR analyses revealed that the amplification was not as successful as expected which suggested that the amplification is affected by presence of inhibitors co-extracted from plant materials. Various potential issues were assessed during DNA extraction and optimisations were made accordingly. A DNA barcoding protocol for O. tenuiflorum was published in the British Pharmacopoeia 2016, which included the reference sequence. The trnH-psbA BPNARM accelerated degradation test which investigates the stability of the reference material over time demonstrated that it has been stable when stored at 56 °C for a year. Using this protocol and trnH-psbA reference material provides a fast and accurate method for identification of O. tenuiflorum. The optimisations of the DNA extraction using the trnH-psbA BPNARM provided a signposting method which can assist in overcoming common problems encountered when using molecular methods with medicinal plants.

Keywords: degradation, DNA extraction, nucleic acid reference material, trnH-psbA

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26 Biosynthesis of Tumor Inhibitory Podophyllotoxin, Quercetin and Kaempferol from Callogenesis of Dysosma Pleiantha (Hance) Woodson

Authors: Palaniyandi Karuppaiya, Hsin Sheng Tsay, Fang Chen

Abstract:

Medicinal herbs do represent a huge and noteworthy reservoir for novel anticancer drugs discovery. Dysosma pleiantha (Hance) Woodson (Berberidaceae), one of the oldest traditional Chinese medicinal herb, highly prized by the mountain tribes of Taiwan and China for its medicinal properties contained pharmaceutically important antitumor compounds podophyllotoxin, quercetin and kaempferol. Among lignans, podophyllotoxin is an active antitumor compound and has now been modified to produce clinically useful drugs etoposide and teniposide. In recent years, natural populations of D. peliantha have declined considerably due to anthropogenic activities such as habitat destruction and commercial exploitation for medicinal applications. As to its overall conservation status, D. pleiantha has been ranked as threatened on the China Species Red List. In the present study, an efficient in vitro callus culture system of D. pleiantha was established on Gamborg’s medium with various combinations and concentrations of different auxins and cytokinins under dark condition. Best callus induction was recorded in 2 mg/L 2, 4 - Dichlorophenoxyacetic acid (2,4-D) along with 0.2 mg/L kinetin and the maximum callus proliferation was achieved at 1 mg/L 2,4-D. Among the explants tested, maximum callus induction (86 %) was achieved from tender leaves. Hence, in subsequent experiments, leaf callus was further investigated for suitable callus biomass and production level of anticancer compounds under the influence of different additives. A maximum fresh callus biomass (8.765 g) was recorded in callus proliferation medium contained 500 mg/L casein hydrolysate. High performance liquid chromatography results revealed that the addition of different concentrations of peptone (1, 2 and 4 g/L) in callus proliferation medium enhanced podophyllotoxin (16 fold), quercetin (12 fold) and kaempferol (5 fold) accumulation than control. Thus, the established in vitro callus culture under the influence of different additives may offer an alternative source of enhanced production of podophyllotoxin, kaempferol and quecertin without harming natural plant population.

Keywords: dysosma pleiantha, kaempferol, podophyllotoxin, quercetin

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25 Synergistic and Antagonistic Interactions between Garlic Extracts and Metformin in Diabetes Treatment

Authors: Ikram Elsiddig, Yacouba Djamila, Amna Hamad

Abstract:

Abstract—The worldwide increasing of using herbs in form of medicine with or without prescription medications potentiates the interactions between herbal products and conventional medicines; due to more research for herb-drug interactions are needed. for a long time hyperglycemia had been treated with several medicinal plants. A. sativum, belonging to the Liliaceae family is well known for its medicinal uses in African traditional medicine, it used for treating of many human diseases mainly diabetes, high cholesterol and high blood pressure. The purpose of this study is to determine the interaction effect between A. sativum bulb extracts and metformin drug used in diabetes treatment. The in vitro and in vivo evaluation were conducted by glucose reuptake using isolated rats hemidiaphgrams tissue and by estimate glucose tolerance in glucose-loaded wistar albino rats. The results showed that, petroleum ether, chloroform and ethyl acetate extracts were found to have activity of glucose uptake in isolated rats hemidiaphgrams of 24.11 mg/g, 19.07 mg/g and 15.66 mg/g compared to metformin drug of 17 mg/g. These activity were reducded to 17.8 mg/g, 13.59 mg/g and 14.46 mg/g after combination with metformin, metformin itself reduced to 13.59 mg/g, 14.46 mg/g and 12.71 mg/g in comination with chloroform and ethyl acetate. These decrease in activity could be due to herbal–drug interaction between the extracts of A. sativum bulb and metformin drug. The interaction between A. sativum extract and metformin was also shown by in vivo study on the induced hyperglycemic rats. The glucose level after administered of 200 mg/kg was found to be increase with 47.2 % and 17.7% at first and second hour compared to the increase of blood glucose in the control group of 82.6% and76.7%.. At fourth hour the glucose level was became less than normal with 3.4% compared to control which continue to increase with 68.2%. Dose of 400 mg/kg at first hour showed increase in blood glucose of 31.5 %, at second and fourth hours the glucose level was became less than normal with decrease of 3.2 % and 30.4%. After combination the activity was found to be less than that of extract at both high and low dose, whereas, at first and second hour, the glucose level was found to be increase with 50.4% and 21.2%, at fourth hour the glucose level was became less than normal with 14%. Therefore A. sativum could be a potential source for anti-diabetic when it used alone, and it is significant important to use the garlic extract alone instead of combined with Metformin drug in diabetes- treatment.

Keywords: Antagonistic, Garlic, Metformin, Synergistic

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24 Efficacy and Safety of Eucalyptus for Relief Cough Symptom: A Systematic Review and Meta-Analysis

Authors: Ladda Her, Juntip Kanjanasilp, Ratree Sawangjit, Nathorn Chaiyakunapruk

Abstract:

Cough is the common symptom of the respiratory tract infections or non-infections; the duration of cough indicates a classification and severity of disease. Herbal medicines can be used as the alternative to drugs for relief of cough symptoms from acute and chronic disease. Eucalyptus was used for reducing cough with evidences suggesting it has an active role in reduction of airway inflammation. The present study aims to evaluate efficacy and safety of eucalyptus for relief of cough symptom in respiratory disease. Method: The Cochrane Library, MEDLINE (PubMed), Scopus, CINAHL, Springer, Science direct, ProQuest, and THAILIS databases. From its inception until 01/02/2019 for randomized control trials. We follow for the efficacy and safety of eucalyptus for reducing cough. Methodological quality was evaluated by using the Cochrane risk of bias tool; two reviewers in our team screened eligibility and extracted data. Result: Six studies were included for the review and five studies were included in the meta-analysis, there were 1.911 persons including children (n: 1) and adult (n: 5) studies; for study in children and adult were between 1 and 80 years old, respectively. Eucalyptus was used as mono herb (n: 2) and in combination with other herbs form (n: 4). All of the studies with eucalyptus were compared for efficacy and safety with placebo or standard treatment, Eucalyptus dosage form in studies included capsules, spray, and syrup. Heterogeneity was 32.44 used random effect model (I² = 1.2%, χ² = 1.01; P-value = 0.314). The efficacy of eucalyptus was showed a reduced cough symptom statistically significant (n = 402, RR: 1.40, 95%CI [1.19, 1.65], P-value < 0.0001) when compared with placebo. Adverse events (AEs) were reported mild to moderate intensity with mostly gastrointestinal symptom. The methodological quality of the included trials was overall poor. Conclusion: Eucalyptus appears to be beneficial and safe for relieving in respiratory diseases focus on cough frequency. The evidence was inconclusive due to limited quality trial. Well-designed trials for evaluating the effectiveness in humans, the effectiveness for reducing cough symptom in human is needed. Eucalyptus had safety as monotherapy or in combination with other herbs.

Keywords: cough, eucalyptus, cineole, herbal medicine, systematic review, meta-analysis

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23 Effect of Labisia pumila var. alata with a Structured Exercise Program in Women with Polycystic Ovarian Syndrome

Authors: D. Maryama AG. Daud, Zuliana Bacho, Stephanie Chok, DG. Mashitah PG. Baharuddin, Mohd Hatta Tarmizi, Nathira Abdul Majeed, Helen Lasimbang

Abstract:

Lifestyle, physical activity, food intake, genetics and medication are contributing factors for people getting obese. Which in some of the obese people were a low or non-responder to exercise. And obesity is very common clinical feature in women affected by Polycystic Ovarian Syndrome (PCOS). Labisia pumila var. alata (LP) is a local herb which had been widely used by Malay women in treating menstrual irregularities, painful menstruation and postpartum well-being. Therefore, this study was carried out to investigate the effect of LP with a structured exercise program on anthropometric, body composition and physical fitness performance of PCOS patients. By using a single blind and parallel study design, where by subjects were assigned into a 16-wk structured exercise program (3 times a week) interventions; (LP and exercise; LPE, and exercise only; E). All subjects in the LPE group were prescribed 200mg LP; once a day, for 16 weeks. The training heart rate (HR) was monitored based on a percentage of the maximum HR (HRmax) achieved during submaximal exercise test that was conducted at wk-0 and wk-8. The progression of aerobic exercise intensity from 25–30 min at 60 – 65% HRmax during the first week to 45 min at 75–80% HRmax by the end of this study. Anthropometric (body weight, Wt; waist circumference, WC; and hip circumference, HC), body composition (fat mass, FM; percentage body fat, %BF; Fat Free Mass, FFM) and physical fitness performance (push up to failure, PU; 1-minute Sit Up, SU; and aerobic step test, PVO2max) were measured at wk-0, wk-4, wk-8, wk-12, and wk-16. This study found that LP does not have a significant effect on body composition, anthropometric and physical fitness performance of PCOS patients underwent a structured exercise program. It means LP does not improve exercise responses of PCOS patients towards anthropometric, body composition and physical fitness performance. The overall data shows exercise responses of PCOS patients is by increasing their aerobic endurance and muscle endurance performances, there is a significant reduction in FM, PBF, HC, and Wt significantly. Therefore, exercise program for PCOS patients have to focus on aerobic fitness, and muscle endurance.

Keywords: polycystic ovarian syndrome, Labisia pumila var. alata, body composition, aerobic endurance, muscle endurance, anthropometric

Procedia PDF Downloads 183
22 Phytochemical Screening and Anti-Hypothyroidism Activity of Lepidium sativum Ethanolic Extract

Authors: Reham Hajomer, Ikram Elsiddig, Amna Hamad

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Lepidium sativum (Garden Cress) belonging to Brassicaceae family is an annual herb locally known as El-rshad. In Ayurveda it is an important medicinal plant, traditionally used for the treatment of jaundice, liver problems, spleen diseases, gastrointestinal disorders, menstrual problems, fracture, arthritis, inflammatory conditions and for treatment of hypothyroidism. Hypothyroidism is a condition in which the thyroid gland does not produce enough thyroid hormones (Triiodithyronine T3 and Thyroxine T4) which are commonly caused by iodine deficiency. It’s divided into primary and secondary hypothyroidism, the primary caused by failure of thyroid function and secondary due to the failure of adequate thyroid-stimulating hormone (TSH) secretion from the pituitary gland or thyroid -releasing hormone (TRH) from the hypothalamus. The disease is most common in women over age 60. The objective regarding this study is to know whether Lepidium sativum would affect the level of thyroid hormones. The extract was prepared with 96% ethanol using Soxhlet apparatus. The anti-hypothyroidism activity was tested by using thirty male Wistar rats weighing (100-140 g) were used in the experiment. They were grouping into five groups, Group 1: Normal group= Administered only distilled water. Then 10 mg/kg Propylthiouracil was added to the drinking water of all other groups to induce hypothyroidism. Group 2: Negative control without any treatment; Group 3: Test group= treated with oral administration of 500mg/kg extract; Group 4: treated with oral administration of 250mg/kg of the extract; Group 5: Standard group (positive control) = treated with intraperitoneal Levothyroxine. All rats were incubated for 20 days at animal house with room temperature of proper ventilation provided with standard diet. The result show that the Lepidium sativum extract was found to increases the T3 and T4 in the propylthiouracil induced rats with values (0.29 ng/dl T3 and 0.57 U T4) for the 500mg/kg and (0.27 ng/dl T3 and 0.517 U T4) for the 250mg/kg in comparison with standard with values (0.241 ng/dl T3 and 0.516 U T4) so that Lepidium sativum can be stimulatory to thyroid function and possess significant anti-hypothyroidism effect with p-values ranges from (0.000006*-0.893472). In conclusion, from results obtained, Lepidium sativum plant extract was found to posses anti-hypothyroidism effects so its act as an agent that stimulates thyroid hormone secretion.

Keywords: anti-hypothyroidism, extract, lepidium, sativum

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21 Being Reticent for Healing – Singularity and Non-Verbalization in Indigenous Medical Practices in Sri Lanka

Authors: Ayami Umemura

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The purpose of this paper is to examine the meaning of verbalization in clinical practice using the keywords silence and singularity. A patient's experience of illness and treatment is singular, irreplaceable, and irreproducible and ultimately cannot be compared with that of others. In his book Difference and Repetition, Gilles Deleuze positioned irreplaceable singularity as the opposite concept of particularity as a generalizable and substitutable property and matched the former with universality. He also said that singularity could not be represented because of its irreplaceable nature. Representation or verbalization is a procedure that converts an irreplaceable, idiosyncratic reality into something that can be substituted. Considering the act of verbalizing medical diagnosis based on this, it can be said that diagnosis is the practice of decontextualizing and generalizing the suffering embedded in the patient's irreplaceable life history as a disease. This paper examines the above with the key concept of the practice of "non-verbalization" in traditional medical practices in Sri Lanka. In the practice of Sri Lankan traditional medicine and the inheritance of medical knowledge and care techniques, there is a tendency to avoid verbalizing specific matters or stating them aloud. Specifically, the following should be avoided. The healer informs the patient of the name of the disease, mentions the name of the herb used in front of the patient, explains the patient's condition to the healer, and referring the names of poisonous animals, such as poisonous snakes that have been damaged. And so on. Furthermore, when passing on medical knowledge and skills, it is also possible to avoid verbalizing knowledge of medicinal herbs and medical treatment methods and explaining them verbally. In addition to the local belief that the soul of language in Sri Lanka is deeply involved in this background, Sri Lankan traditional medicine has a unique view of the human body and personality that is rooted in the singularity that appears in the relationship with the movement of celestial bodies and the supernatural realm. It can be pointed out that it is premised on the view. In other words, the “silence” in Sri Lankan indigenous medicine is the reason for emphasizing specificity. Furthermore, we can say that "non-verbalization" is a practice aimed at healing. Based on these discussions, this paper will focus on the unique relationships between practitioners and patients that become invisible due to verbalization, which is overlooked by clinical medicine, where informed consent, ensuring transparency, and audit culture is dominant. We will examine the experience of treatment and aim to relativize clinical medicine, which is based on audit cultures.

Keywords: audit cultures, indigenous medicine, singularity, verbalization

Procedia PDF Downloads 63
20 Comparative Therapeutic Potential of 'Green Synthesized' Antimicrobials against Scalp Infections

Authors: D. Desai, J.Dixon, N. Jain, M. Datta

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Microbial infections of scalp consist of symptomatic appearances associated with seborrhoeic dermatitis, folliculitis, furuncles, carbuncles and ringworm. The main causative organisms in these scalp-based infections are bacteria like S. aureus, P. aeruginosa and a fungus M. Furfur. Allopathic treatment of these infections is available and efficient, but occasionally, topical applications have been found to cause side effects. India is known as the botanical garden of the world and considered as the epicentre for utilization of traditional drugs. Many treatments based on herb extracts are commonly used in India. It has been observed treatment with ethnomedicines requires a higher dosage and greater time period. Additionally, repeated applications are required to obtain the full efficacy of the treatment. An attempt has been made to imbibe the traditional knowledge with nanotechnology to generate a proficient therapeutic against scalp infections. We have imbibed metallic nanoparticles with extracts from traditional medicines and propose to formulate an antimicrobial hair massager. Four commonly used herbs for treatment against scalp disorders like Zingiber officinale (ginger), Allium sativum (garlic), Azadirachta indica (neem) leaves and Citrus limon (lemon) peel was taken. 30 gms of dried homogenized powder was obtained and processed for obtaining the aqueous and ethanolic extract by soxhlet apparatus. The extract was dried and reconstituted to obtain working solution of 1mg/ml. Phytochemical analysis for the obtained extract was done. Synthesis of nanoparticles was mediated by incubating 1mM silver nitrate with extracts of various herbs to obtain silver nanoparticles. The formation of the silver nanoparticles (AgNPs) was monitored using UV-Vis spectroscopy. The AgNPs thus obtained were centrifuged and dried. The AgNPs thus formed were characterized by X Ray Diffraction, scanning electron microscopy and transmission electron microscopy. The size of the AgNPs varied from 10-20 nm and was spherical in shape. P. aeruginosa was plated on nutrient agar and comparative antibacterial activity was tested. Comparative antimicrobial potential was calculated for the extracts and the corresponding nanoconstructs. It was found AgNPs were more efficient than their aqueous and ethanolic counterparts except in the ase of C. limon. Statistical analysis was performed to validate the results obtained.

Keywords: ethnomedicine, nanoconstructs, scalp infections, Zingiber officinale

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19 Insectivorous Medicinal Plant Drosera Ecologyand its Biodiversity Conservation through Tissue Culture and Sustainable Biotechnology

Authors: Sushil Pradhan

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Biotechnology contributes to sustainable development in several ways such as biofertilizer production, biopesticide production and management of environmental pollution, tissue culture and biodiversity conservation in vitro, in vivo and in situ, Insectivorous medicinal plant Drosera burmannii Vahl belongs to the Family-Droseraceae under Order-Caryophyllales, Dicotyledoneae, Angiospermeae which has 31 (thirty one) living genera and 194 species besides 7 (seven) extinct (fossil) genera. Locally it is known as “Patkanduri” in Odia. Its Hindi name is “Mukhajali” and its English name is “Sundew”. The earliest species of Drosera was first reported in 1753 by Carolous Linnaeus called Drosera indica L (Indian Sundew). The latest species of Drosera reported by Fleisch A, Robinson, AS, McPherson S, Heinrich V, Gironella E and Madulida D.A. (2011) is Drosera ultramafica from Malaysia. More than 50 % species of Drosera have been reported from Australia and next to Australia is South Africa. India harbours only 3 species such as D. indica L, Drosera burmannii Vahl and D. peltata L. From our Odisha only D. burmannii Vahl is being reported for the first time from the district of Subarnapur near Sonepur (Arjunpur Reserve Forest Area). Drosera plant is autotrophic but to supplement its Nitrogen (N2) requirement it adopts heterotrophic mode of nutrition (insectivorous/carnivorous) as well. The colour of plant in mostly red and about 20-30cm in height with beautiful pink or white pentamerous flowers. Plants grow luxuriantly during November to February in shady and moist places near small water bodies of running water stream. Medicinally it is a popular herb in the locality for the treatment of cold and cough in children in rainy season by the local Doctors (Kabiraj and Baidya). In the present field investigation an attempt has been made to understand the unique reproductive phase and life cycle of the plant thereby planning for its conservation and propagation through various techniques of tissue culture and biotechnology. More importantly besides morphological and anatomical studies, cytological investigation is being carried out to find out the number of chromosomes in the cell and its genomics as there is no such report as yet for Drosera burmannii Vahl. The ecological significance and biodiversity conservation of Drosera with special reference to energy, environmental and chemical engineering has been discussed in the research paper presentation.

Keywords: insectivorous, medicinal, drosera, biotechnology, chromosome, genome

Procedia PDF Downloads 357
18 A Potential Bio-Pesticidal Molecule Derived from Indian Traditional Plant

Authors: Bunindro Nameirakpam, Sonia Sougrapakam, Shannon B. Olsson, Rajashekar Yallappa

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Natural sources for new pesticidal compounds hold promise in view of their eco-friendly nature, selectivity and mammalian safety. Despite a large number of plants that show insecticidal activity and diversity of natural chemistry with inherent eco-friendly nature, newer classes of insecticides have eluded discovery. Artemisia vulgaris, known as Mugwort, is a universal herb used for folk medicine and religious purposes throughout the ancient world. In India, the essential oils of Artemisia vulgaris are used for its insecticidal, anti parasiticidal and antimicrobial properties. Traditionally, the dried leaves of Artemisia vulgaris are used to repel insects as well as rats in and around the granaries in the North-East India. Artemisia vulgaris collected during November from different ecological sites were studied for the bio-pesticidal utility against the stored grain pests. The insecticidal activities were found in the crude extracts of n-hexane and methanol from the samples collected in Sikkim and Manipur respectively. Using silica gel column chromatography protocol, we have isolated one novel bioactive molecule from the aerial parts of Artemisia vulgaris L based on various physical-chemical and spectroscopic techniques (IR, 1H NMR, 13C NMR and mass). The novel bioactive molecule is highly toxic and very low concentration (4.35 µg/l) is needed to control the stored product insects. In additional experiment results clearly showed the involvement of sodium pumps inhibition in the insecticidal action of purified compound in the Sitophilus oryzae. The knockdown activity of the purified compound is concomitant with the in vivo inhibition of Na+/ K+- ATPase. Further, our study showed insignificant differences in the seed germination of control and the treated grains. The lack of adverse effect of the novel bioactive molecule on the seed germination is highly desirable for seed/grain protectant and showing the potential to be developed as possible natural fumigants for the control of stored grain pests. The novel bioactive molecule is selective insecticide with a high margin of safety to mammals and showed promise as novel biopesticide candidate for grain protection. It is believed that Bio-pesticides can serve as the most important pest management tools as far as global safety is concerned.

Keywords: Indian traditional plant, Artemisia vulgaris, bio-pesticides, Na+/ K+- ATPase, seed germination

Procedia PDF Downloads 165
17 Prophylactic Effect of Dietary Garlic (Allium sativum) Inclusion in Feed of Commercial Broilers with Coccidiosis Raised at the Experimental Animal Unit of the Department of Veterinary Medicine, University of Ibadan, Oyo State, Nigeria

Authors: Ogunlesi Olufunso, John Ogunsola, Omolade Oladele, Benjamin Emikpe

Abstract:

Context: Coccidiosis is a parasitic disease that affects poultry production, leading to economic losses. Garlic is known for medicinal properties and has been used as a natural remedy for various diseases. This study aims to investigate the prophylactic effect of garlic inclusion in the feed of commercial broilers with coccidiosis. Research Aim: The aim of this study is to determine the possible effect of garlic meal inclusion in poultry feed on the body weight gain of commercial broilers and to investigate it's therapeutic effect on broilers with coccidiosis. Methodology: The study conducted a case-control study for eight weeks with One hundred Arbor acre commercial broilers separated into five (5) groups from day-old, where 6,000 Eimeria oocysts were orally inoculated into each broiler in the different groups. Feed intake, body weight gain, feed conversion ratio, oocyt shedding rate, histopathology and erythrocyte indices were assessed. Findings: The inclusion of garlic meal in the broilers' diet resulted in an improved feed conversion ratio, decreased oocyst counts, reduced diarrhoeic fecal spots, decreased susceptibility to coccidial infection, and increased packed cell volume (PCV). Theoretical Importance: This study contributes to the understanding of the prophylactic effect of garlic supplementation, including its antiparasitic properties on commercial broilers with coccidiosis. It highlights the potential use of non-conventional feed additives or ayurvedic herb and spices in the treatment of poultry diseases. Data Collection and Analysis Procedures: The study collected data on feed intake, body weight gain, oocyst shedding rate, histopathological observations, and erythrocyte indices. Data were analyzed using Analysis of Variance and Duncan's Multiple range Test. Questions Addressed: The study addressed the possible effect of garlic meal inclusion in poultry feed on the body weight gain of broilers and its therapeutic effect on broilers with coccidiosis. Conclusion: The study concludes that garlic inclusion in the feed of broilers has a prophylactic effect, including antiparasitic properties, resulting in improved feed conversion ratio, reduced oocyst counts and increased PCV.

Keywords: broilers, eimeria spp, garlic, Ibadan

Procedia PDF Downloads 44
16 An Efficient and Low Cost Protocol for Rapid and Mass in vitro Propagation of Hyssopus officinalis L.

Authors: Ira V. Stancheva, Ely G. Zayova, Maria P. Geneva, Marieta G. Hristozkova, Lyudmila I. Dimitrova, Maria I. Petrova

Abstract:

The study describes a highly efficient and low-cost protocol for rapid and mass in vitro propagation of medicinal and aromatic plant species (Hyssopus officinalis L., Lamiaceae). Hyssop is an important aromatic herb used for its medicinal values because of its antioxidant, anti-inflammatory and antimicrobial properties. The protocol for large-scale multiplication of this aromatic plant was developed using young stem tips explants. The explants were sterilized with 0.04% mercuric chloride (HgCl₂) solution for 20 minutes and washing three times with sterile distilled water in 15 minutes. The cultural media was full and half strength Murashige and Skoog medium containing indole-3-butyric acid. Full and ½ Murashige and Skoog media without auxin were used as controls. For each variant 20 glass tubes with two plants were used. In each tube two tip and nodal explants were inoculated. Maximum shoot and root number were obtained on ½ Murashige and Skoog medium supplemented with 0.1 mg L-1 indole-3-butyric acid at the same time after four weeks of culture. The number of shoots per explant and shoot height were considered. The data on rooting percentage, the number of roots per plant and root length were collected after the same cultural period. The highest percentage of survival 85% for this medicinal plant was recorded in mixture of soil, sand and perlite (2:1:1 v/v/v). This mixture was most suitable for acclimatization of all propagated plants. Ex vitro acclimatization was carried out at 24±1 °C and 70% relative humidity under 16 h illuminations (50 μmol m⁻²s⁻¹). After adaptation period, the all plants were transferred to the field. The plants flowered within three months after transplantation. Phenotypic variations in the acclimatized plants were not observed. An average of 90% of the acclimatized plants survived after transferring into the field. All the in vitro propagated plants displayed normal development under the field conditions. Developed in vitro techniques could provide a promising alternative tool for large-scale propagation that increases the number of homologous plants for field cultivation. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: Hyssopus officinalis L., in vitro culture, micro propagation, acclimatization

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15 The Efficacy of Thymbra spicata Ethanolic Extract and its Main Component Carvacrol on In vitro Model of Metabolically-Associated Dysfunctions

Authors: Farah Diab, Mohamad Khalil, Francesca Storace, Francesca Baldini, Piero Portincasaa, Giulio Lupidi, Laura Vergani

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Thymbra spicata is a thyme-like plant belonging to the Lamiaceae family that shows a global distribution, especially in the eastern Mediterranean region. Leaves of T. spicata contain large amounts of phenols such as phenolic acids (rosmarinic acid), phenolic monoterpenes (carvacrol), and flavonoids. In Lebanon, T. spicata is currently used as a culinary herb in salad and infusion, as well as for traditional medicinal purposes. Carvacrol (5-isopropyl-2-methyl phenol), the most abundant polyphenol in the organic extract and essential oils, has a great array of pharmacological properties. In fact, carvacrol is largely employed as a food additive and neutraceutical agent. Our aim is to investigate the beneficial effects of T. spicata ethanolic extract (TE) and its main component, carvacrol, using in vitro models of hepatic steatosis and endothelial dysfunction. As a further point, we focused on investigating if and how the binding of carvacrol to albumin, the physiological transporter for drugs in the blood, might be altered by the presence of high levels of fatty acids (FAs), thus impairing the carvacrol bio-distribution in vivo. For that reason, hepatic FaO cells treated with exogenous FAs such as oleate and palmitate mimic hepatosteatosis; endothelial HECV cells exposed to hydrogen peroxide are a model of endothelial dysfunction. In these models, we measured lipid accumulation, free radical production, lipoperoxidation, and nitric oxide release before and after treatment with carvacrol. The carvacrol binding to albumin with/without high levels of long-chain FAs was assessed by absorption and emission spectroscopies. Our findings show that both TE and carvacrol (i) counteracted lipid accumulation in hepatocytes by decreasing the intracellular and extracellular lipid contents in steatotic FaO cells; (ii) decreased oxidative stress in endothelial cells by significantly reducing lipoperoxidation and free radical production, as well as, attenuating the nitric oxide release; (ii) high levels of circulating FAs reduced the binding of carvacrol to albumin. The beneficial effects of TE and carvacrol on both hepatic and endothelial cells point to a nutraceutical potential. However, high levels of circulating FAs, such as those occurring in metabolic disorders, might hinder the carvacrol transport, bio-distribution, and pharmacodynamics.

Keywords: carvacrol, endothelial dysfunction, fatty acids, non-alcoholic fatty liver diseases, serum albumin

Procedia PDF Downloads 154
14 Evaluation of Medicinal Plants, Catunaregam spinosa, Houttuynia cordata, and Rhapis excelsa from Malaysia for Antibacterial, Antifungal and Antiviral Properties

Authors: Yik Sin Chan, Bee Ling Chuah, Wei Quan Chan, Ri Jin Cheng, Yan Hang Oon, Kong Soo Khoo, Nam Weng Sit

Abstract:

Traditionally, medicinal plants have been used to treat different kinds of ailments including infectious diseases. They serve as a good source of lead compounds for the development of new and safer anti-infective agents. This study aimed to investigate the antimicrobial potential of the leaves of three medicinal plants, namely Catunaregam spinosa (Rubiaceae; Mountain pomegranate), Houttuynia cordata (Saururaceae; "fishy-smell herb") and Rhapis excelsa (Arecaceae; “broadleaf lady palm”). The leaves extracts were obtained by sequential extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and water. The antibacterial and antifungal activities were assessed using a colorimetric broth microdilution method against a panel of human pathogenic bacteria (Gram-positive: Bacillus cereus and Staphylococcus aureus; Gram-negative: Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) and fungi (yeasts: Candida albicans, Candida parapsilosis and Cryptococcus neoformans; Moulds: Aspergillus fumigatus and Trichophyton mentagrophytes) respectively; while antiviral activity was evaluated against the Chikungunya virus on monkey kidney epithelial (Vero) cells by neutral red uptake assay. All the plant extracts showed bacteriostatic activity, however, only 72% of the extracts (13/18) were found to have bactericidal activity. The lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were given by the hexane extract of C. spinosa against S. aureus with the values of 0.16 and 0.31 mg/mL respectively. All the extracts also possessed fungistatic activity. Only the hexane, chloroform and ethyl acetate extracts of H. cordata exerted inhibitory activity against A. fumigatus, giving the lowest fungal susceptibility index of 16.7%. In contrast, only 61% of the extracts (11/18) showed fungicidal activity. The ethanol extract of R. excelsa exhibited the strongest fungicidal activity against C. albicans, C. parapsilosis and T. mentagrophytes with minimum fungicidal concentration (MFC) values of 0.04–0.08 mg/mL, in addition to its methanol extract against T. mentagrophytes (MFC=0.02 mg/mL). For anti-Chikungunya virus activity, only chloroform and ethyl acetate extracts of R. excelsa showed significant antiviral activity with 50% effective concentrations (EC50) of 29.9 and 78.1 g/mL respectively. Extracts of R. excelsa warrant further investigations into their active principles responsible for antifungal and antiviral properties.

Keywords: bactericidal, Chikungunya virus, extraction, fungicidal

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13 Antimicrobial Activity of Igusa and the Application to Foam Materials for Food Industry

Authors: I. Nanako, Mariko Era, Hiroshi Morita

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Objectives: Japanese uses TATAMI rather than flooring at home. Igusa ( Juncus effuses var. decipiens ), which is commonly known in the forms of TATAMI. Juncus spp. grow at a relatively high humidity area (Japan, China and Southeast Asia ). Yatsushiro region in the southern part of Kumamoto prefecture is major produing area of Igusa. Igusa found to have honeycomb structure and was also shown to have the ability to control humidity. And Igusa has been used as a medicinal herb for diuretic and antiphlogistic agent. In previous study, we investigated antimicrobial effects of Igusa, and showed high antimicrobial activity against food poisoning bacteria. Therefore, the food trays blended Igusa can be kept clean by antimicrobial activity of Igusa. We focus on ‘Igusa foam materials’. In this study, we investigated the antibacterial and antifungal activity of Igusa, and new application to foam materials for food industry. Materials and method: We used Igusa foam materials (3 × 3 × 3 cm) as a sample. We set about fifteen types of samples combined with a commercial antibacterial agent A, a commercial antibacterial agent B, potassium laurate (C12K) and a commercial antifungal agent C, a commercial antifungal agent D and a commercial antifungal agent E. We selected four bacteria strains (Escherichia coli NBRC 3972, Staphylococus aureus NBRC 12732, Salmonella typhimurium NBRC 13245, Bacillus subtilis NBRC 3335 ) and three fungus strains (Penicillium pinophilum NBRC 6345, Cladosporium cladosporioides NBRC 30314, Aspergillus oryzae NBRC 5238 ). The fungus was cultured at 30 °C on Igusa foam materials after inoculation of the fungus for fourteen days. The bacteria was cultured at 30 °C on Igusa foam materials after inoculation of the bacteria for three days. And the Igusa foam materials were washed with 10 mL normal saline after three days. The normal saline washed Igusa foam materials plated the NA medium. After, It was cultured at 30 °C and used colony counting method. Result and Conclusion: The fifteen types of sample of Igusa foam materials had antifungal activity against C. cladosporioides, A. oryzae and P. pinophilum for fourteen days. The four types of sample contained potassium laurate and antibacterial agent A, sample contained antibacterial agent B and antifungal agent D, sample contained A and antifungal agent E, sample contained B and E had antibacterial activity against B. subtilis. The three types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E had antibacterial activity against S. typhimurium. The five types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E, sample contained B and E, sample contained B and antifungal agent C had antibacterial activity against E. coli and S. aureus. These results indicate that Igusa of Igusa foam materials had high antifungal activity. In addition, Igusa foam materials combined with a commercial antibacterial agent had antibacterial activity. In the future, we consider that use of Igusa foam materials may be spread from food industry.

Keywords: antibacterial, antifungal, foam materials, Igusa

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12 Effect of Plant Growth Regulators on in vitro Biosynthesis of Antioxidative Compounds in Callus Culture and Regenerated Plantlets Derived from Taraxacum officinale

Authors: Neha Sahu, Awantika Singh, Brijesh Kumar, K. R. Arya

Abstract:

Taraxacum officinale Weber or dandelion (Asteraceae) is an important Indian traditional herb used to treat liver detoxification, digestive problems, spleen, hepatic and kidney disorders, etc. The plant is well known to possess important phenolic and flavonoids to serve as a potential source of antioxidative and chemoprotective agents. Biosynthesis of bioactive compounds through in vitro cultures is a requisite for natural resource conservation and to provide an alternative source for pharmaceutical applications. Thus an efficient and reproducible protocol was developed for in vitro biosynthesis of bioactive antioxidative compounds from leaf derived callus and in vitro regenerated cultures of Taraxacum officinale using MS media fortified with various combinations of auxins and cytokinins. MS media containing 0.25 mg/l 2, 4-D (2, 4-Dichloro phenoxyacetic acid) with 0.05 mg/l 2-iP [N6-(2-Isopentenyl adenine)] was found as an effective combination for the establishment of callus with 92 % callus induction frequency. Moreover, 2.5 mg/l NAA (α-Naphthalene acetic acid) with 0.5 mg/l BAP (6-Benzyl aminopurine) and 1.5 mg/l NAA showed the optimal response for in vitro plant regeneration with 80 % regeneration frequency and rooting respectively. In vitro regenerated plantlets were further transferred to soil and acclimatized. Quantitative variability of accumulated bioactive compounds in cultures (in vitro callus, plantlets and acclimatized) were determined through UPLC-MS/MS (ultra-performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry) and compared with wild plants. The phytochemical determination of in vitro and wild grown samples showed the accumulation of 6 compounds. In in vitro callus cultures and regenerated plantlets, two major antioxidative compounds i.e. chlorogenic acid (14950.0 µg/g and 4086.67 µg/g) and umbelliferone (10400.00 µg/g and 2541.67 µg/g) were found respectively. Scopoletin was found to be highest in vitro regenerated plants (83.11 µg/g) as compared to wild plants (52.75 µg/g). Notably, scopoletin is not detected in callus and acclimatized plants, but quinic acid (6433.33 µg/g) and protocatechuic acid (92.33 µg/g) were accumulated at the highest level in acclimatized plants as compared to other samples. Wild grown plants contained highest content (948.33 µg/g) of flavonoid glycoside i.e. luteolin-7-O-glucoside. Our data suggests that in vitro callus and regenerated plants biosynthesized higher content of antioxidative compounds in controlled conditions when compared to wild grown plants. These standardized cultural conditions may be explored as a sustainable source of plant materials for enhanced production and adequate supply of oxidative polyphenols.

Keywords: anti-oxidative compounds, in vitro cultures, Taraxacum officinale, UPLC-MS/MS

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11 Arbuscular Mycorrhizal Symbiosis Modulates Antioxidant Capacity of in vitro Propagated Hyssop, Hyssopus officinalis L.

Authors: Maria P. Geneva, Ira V. Stancheva, Marieta G. Hristozkova, Roumiana D. Vasilevska-Ivanova, Mariana T. Sichanova, Janet R. Mincheva

Abstract:

Hyssopus officinalis L., Lamiaceae, commonly called hyssop, is an aromatic, semi-evergreen, woody-based, shrubby perennial plant. Hyssop is a good expectorant and antiviral herb commonly used to treat respiratory conditions such as influenza, sinus infections, colds, and bronchitis. Most of its medicinal properties are attributed to the essential oil of hyssop. The study was conducted to evaluate the influence of inoculation with arbuscular mycorrhizal fungi of in vitro propagated hyssop plants on the: activities of antioxidant enzymes superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase; accumulation of non-enzymatic antioxidants total phenols and flavonoid, water-soluble soluble antioxidant metabolites expressed as ascorbic acid; the antioxidant potential of hyssop methanol extracts assessed by two common methods: free radical scavenging activity using free stable radical (2,2-diphenyl-1-picrylhydrazyl, DPPH• and ferric reducing antioxidant power FRAP in flowers and leaves. The successfully adapted to field conditions in vitro plants (survival rate 95%) were inoculated with arbuscular mycorrhizal fungi (Claroideoglomus claroideum, ref. EEZ 54). It was established that the activities of enzymes with antioxidant capacity (superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase) were significantly higher in leaves than in flowers in both control and mycorrhized plants. In flowers and leaves of inoculated plants, the antioxidant enzymes activity were lower than in non-inoculated plants, only in SOD activity, there was no difference. The content of low molecular metabolites with antioxidant capacity as total phenols, total flavonoids, and water soluble antioxidants was higher in inoculated plants. There were no significant differences between control and inoculated plants both for FRAP and DPPH antioxidant activity. According to plant essential oil content, there was no difference between non-inoculated and inoculated plants. Based on our results we could suggest that antioxidant capacity of in vitro propagated hyssop plant under conditions of cultivation is determined by the phenolic compounds-total phenols and flavonoids as well as by the levels of water-soluble metabolites with antioxidant potential. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: antioxidant enzymes, antioxidant metabolites, arbuscular mycorrhizal fungi, Hyssopus officinalis L.

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10 White Clover Trifolium repens L. Genetic Diversity and Salt Tolerance in Urban Area of Riga

Authors: Dace Grauda, Gunta Cekstere, Inta Belogrudova, Andis Karlsons, Isaak Rashal

Abstract:

Trifolium repens L. (white or Dutch clover) is a perennial herb, belongs to legume family (Leguminosae Juss.), spread extensively by stolons and seeds. The species is cultivated worldwide and was naturalized in many countries in meadows, yards, gardens, along roads and streets etc., especially in temperate regions. It is widespread also in grasslands throughout Riga, the capital of Latvia. The goal of this study was to investigate genetic structure of white clover population in Riga and to evaluate influence of different salt concentration on plants. For this purpose universal retrotranspozone based IRAP (Inter-Retrotransposon Amplified Polymorphism) method was used. The plant material was collected in different regions of Riga and in several urban areas of Latvia. Plant DNA was isolated from in silicogel dried leaves of using 1% CTAB (cetyltrimet-ammonium bromide) buffer DNA extraction procedure. Genetic structure of city population and wild populations were compared. Soil salinization is an important issue associated with low water resources and highly urbanized areas in aride and semi-aride climate conditions, as well as de-icing salt application to prevent ice formation on roads in winter. The T. repens variety ‘Daile’ (form giganteum), one of the often used component of urban greeneries, was studied in this investigation. Plants were grown from seeds and cultivated in the light conditions (18-25 C, 16h/8h of day/night, light intensity 3000 lx) in plastic pots (200 ml), filled with commercial neutralized (pH 5.9 ± 0.3) peat substrate with mineral nutrients. To analyse the impact of increased soil salinity treatments with gradually rising NaCl (0; 20; 40; 60; 80; 100 mM) levels were arranged. Plants were watered when necessary with deionised water to provide optimum substrate moisture 60-70%. The experiment was terminated six weeks after establishment. For analysis of mineral nutrients, dry plant material (above ground part and roots) was used. Decrease of Na content can be significant under elevated salinity till 20 mM NaCl. High NaCl concentrations in the substrate increase Na, Cl, Cu, Fe, and Mn accumulation, but reduce S, Mg, K content in the plant above ground parts. Abiotic stresses generally changes the levels of DNA metilation. Several candidate gene for salt tolerance will be analysed for DNA metilation level using Pyromark-Q24 advanced.

Keywords: DNA metilation, IRAP, soil salinization, white clover

Procedia PDF Downloads 337