Phylogenetic analysis and biological characteristic tests of marine bacteria isolated from Southern Ocean(Indian sector)water

Fifty-seven bacteria were isolated from Southern Ocean （Indian sector） water samples which were collected from different latitude and longitude of the ocean. All the isolates were able to grow at 4℃, 20℃, 37℃ and tolerable NaCI concentration up to 13.5% （w/v）. 29 out of 57 isolates were identified using 16S rDNA amplification and the sequences were submitted to National Center for Biotechnology Information （NCBI）. All the isolates were classified by using Ribosomal Database Project （RDP） and found that isolates belongs to Proteobacteria and Bacteriodes. The average G＋C content was 56.4%. The isolates were screened for the presence of extracellular enzymes, viz. amylase, catalase, urease, esterase, lipase and protease. The disc diffusion method is used to screen antibiotic production by the isolates against four pathogenic bacteria, viz. Salmonella typhimurium （NCIM 2501）, Staphylococcus aureus （NCIM 2122）, Bacillus subtilis （NCIM 2193）, and Pseudomonas aeruginosa （NCIM 2036）. Nine out of 29 were found to be antibiotic producer.

Construction and Identification of the Helper Plasmids for Reverse Genetic System of Rabies Virus Street Strain

To obtain the helper plasmids for a reverse genetics system of rabies virus, the cDNAs of the complete open reading frames of the N, P, G, and L genes of rabies street virus stain HN10 were each cloned into expression vector pVAX1, These four plasmids were identified by restriction enzyme digestion and gene sequencing. The plasmid encoding the N protein was selected to determine the expression effect of these plasmids in NA cells. The results showed that the helper plasmids for a reverse genetics system of rabies street virus strain HN10 had been successfully constructed.

The Effects of Omega-3 Fatty Acids Supplementation in Bronchial Asthma

Bronchial asthma is a chronic inflammatory disease. Allergies are intensely related to bronchial asthma and to other respiratory diseases such as chronic sinusitis, middle ear infections, and nasal polyps. Treatment can vary from inhalers to oral medications to drugs delivered in a nebulizer or breathing machine. Besides, dietary involvement was a significant tool to reduce the severity of many chronic inflammatory diseases including asthma. Randomly assigned, double blind, and placebo controlled 290 adults with mild to moderately persistent bronchial asthma concluded in this study, were subjected to alternating phases of supplementation with omega-3 fatty acids, vitamin C and Zn either singly or in combination. Subjective symptom improvement, pulmonary function, and biochemical tests were carried out at the beginning of the study and at the end of each therapeutic phase. The study findings showed that nearly half of the cases were in age between 18 years to 40 years followed by 29.6 of the cases that fall in age range between 40 years and 60 years. It was clear that all the four supplements (ogema-3 fatty acids, vitamin C and Zn, and combination) contributed more than placebo in reducing the severity of bronchial asthma. However, omega 3 fatty acids and combined supplement significantly contributed in symptomatic improvement (p < 0.05). There was a significant improvement of pulmonary function and sputum inflammatory markers with diet supplementation (p < 0.05). So, the subjects with mild and moderately persistent bronchial asthma may get benefit from their dietary supplementation with omega 3 fatty acids, Zinc and vitamin C. It is evident from this study that omega 3 fatty acids and combined supplementation significantly improved the severity of bronchial asthma.

Ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry-characterized extract of Aerides odorata Lour alleviates paracetamol-induced hepatotoxicity in animal model evidenced by biochemical,molecular,and computational studies

Background:Many kinds of orchids have significant health benefits although ade-quate research on their biological functions is yet to be carried out.This study inves-tigated the paracetamol-induced liver damage-protecting effect of epiphytic Aerides odorata methanol extract(AODE).Methods:The protective effects of AODE were studied by analyzing its effect on liver function parameters,messenger RNA(mRNA)expression,and tissue histopatho-logical architecture.The results were confirmed by ligand-receptor interaction of molecular docking and multitarget interaction of network pharmacological analyses.Results:AODE significantly(p<0.05)minimized the dose-dependent increase in acid phosphatase,aspartate aminotransferase,alanine aminotransferase,alkaline phos-phatase,γ-glutamyl transferase,lactate dehydrogenase,and total bilirubin compared to the reference drug silymarin.Malondialdehyde level decreased,and the antioxidant genes catalase(CAT),superoxide dismutase(SOD),β-actin,paraoxonase-1(PON1),and phosphofructokinase-1(PFK-1)were upregulated in AODE-treated paracetamol-intoxicated rats.A total of 376 compounds comprising phenols and flavonoids were identified using ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry(UPLC-qTOF-MS).The online toxicity assessment using SwissADME and admetSAR exhibited drug-like,nontoxic,and potential pharmaco-logical properties.Additionally,in silico analysis showed that isoacteoside,one of the identified compounds,exhibited the best docking score(−11.42)with the liver pro-tein human pituitary adenylate cyclase-1(Protein Data Bank ID:3N94).Furthermore,network pharmacology analysis identified the top 10 hub genes,namely AKT1(protein kinase B),CTNNB1(catenin beta-1),SRC(proto-oncogene c-Src),TNF(tumor necrosis factor),EGFR(epidermal growth factor receptor),HSP90AA1(heat shock protein 90α),MAPK3(mitogen-activated protein kinase 3),STAT3(signal transducer and activator of transcription 3),CASP3(caspase protein),and ESR1(estrogen receptor 1),which are responsible for hepatoprotective activity.Conclusion:The findings demonstrate that AODE could be a novel hepatoprotective target in drug-induced liver damage with a further single compound-based animal study.

Comparative assessment of total polyphenols, antioxidant and antimicrobial activity of different tea varieties of Bangladesh

Objective: To determine the total polyphenol content, antioxidant activity and antibacterial properties of the extracts of different Bangladeshi tea varieties such as flowery broken orange pekoe, broken orange pekoe, red dust and green tea.Methods: Total phenolic and flavonoid contents were determined by Folin–Ciocalteu and aluminum chloride colorimetric assay, respectively.The antioxidant capacity was determined by ferric ion reducing antioxidant power and phosphomolybdenum method.Antibacterial activity was evaluated by disc diffusion method in agar plate and subsequently, the minimum inhibitory concentration was determined by broth dilution method.Results: Total phenolic and flavonoid contents were significantly higher(P < 0.05) in green tea compared to other three black tea varieties.The green tea also showed a higher free radical scavenging and antioxidant activities than all the other tea varieties tested(P < 0.05).In addition, the extracts of all four tea varieties showed inhibitory activity against several pathogenic bacteria and also the same trend of higher antimicrobial activity of green tea than other tea varieties was observed.Conclusions: Taken together, the results of this study demonstrated that Bangladeshi tea,especially the green tea, may act as a substitute for natural antioxidants and as a promising antibacterial agent for beneficial influence in human health.

In vitro callus induction and plantlet regeneration of Achyranthes aspera L.,a high value medicinal plant

Objective:To study callus induction from different explants(internode,leaf,root)and in vitro plantlets propagation from medicinally important plant Achyranthes aspera L.Methods:Sterilized explants were prepared by uning 0.1%HgCl_2 and 0.5%Bavistin and callus was obtained when cultured onto Murashige Skoog's(MS)medium by using different concentrations and combination of 2,4-D.NAA.BAP,IAA,IBA with 3%sucrose and 0.8%agar.Induced callus was immediately transferred to MS medium containing at different concentrations of phytohormones for shootlets and rootlets induction respectively.Results:Sterilization treatment of 0.1%HgCl_2.for 2-3 min and Bavistin 0.5%for 10-12 min showed the highest percentage of asepsis and survival rate.Maximum induction of callus was obtained from a combination of 2.0 mg/L 2,4-D and 0.5 mg/L NAA from leaf.Highest shootlets number(4.83±0.l7)and length(3.8±0.16)cm were observed on full strength MS medium when fortified with BAP 4.0 mg/L and KIN 0.5 mg/L.Concerted efforts of BAP 10 mg/L and NAA 0.5 mg/L on full strength MS medium showed highest leaf number(6.77±0.94).In vitro raised shoots were allowed to root on different strengths of MS medium fortified with IAA and IBA at different concentrations.Experimentally,3.0 mg/L IBA was enabled to induce maximum rootlets number(10.0±9.82)on full strength MS medium.Afterwards,regenerated shoots with well developed roots were successfully subjected to hardening process and were acclimatized.The survived plantlets showed 66.67%survival frequency without any morphological abnormality.Conclusions:The results demonstrated that different explants were good source of callus induction,morphology analysis as well as indirect plantlets regeneration.

Analysis of immune responses against H pylori in rabbits

AIM： To investigate the immunogenicity of H pylori proteins, to evaluate the production rate of anti H pylori IgG antibodies in relation to time and to demonstrate the fidelity of newly optimized in-house enzymelinked immunosorbent assay （ELISA） technique as an alternative for H pylori infection assay. METHODS： In the present study, 100 μg of formalinfixed H pylori whole cell antigens was injected into an experimental animal （New Zealand white female rabbit） intramuscularly on d 0, 16, 27 and 35. The first two doses were injected with adjuvants. On d 0, a serum sample was collected from the rabbit before immunization and this pre-immunized serum was used as a negative control for the whole study. To evaluate the immunogenic responses of the injected antigen, serum samples were collected from the rabbit at regular intervals up to d 42. The sera were analyzed using inhouse ELISA and Western blot techniques.RESULTS： The production of anti Hpylor/IgG antibodies in the rabbit in response to the injected antigen increased almost exponentially up to d 14 and after that it was maintained at the same level until the last day （d 42）. By analyzing the immune profiles of immunized sera, 11 proteins were identified to be immunogenic, among them 2 （approximately 100 kDa and 85 kDa） were most prominent. CONCLUSION： Analysis of the immune responses against pathogenic microorganisms like H py/ori is necessary for the development of various diagnostic and preventive approaches. The results of this experiment reveal that the formalin-fixed H pylori whole cell antigens injected into the rabbit are highly immunogenic. These prominent proteins （approximately 100 kDa and 85 kDa） might have higher immunogenic effects among humans infected with H pylori and some of these immunogenic proteins can be included in diagnostic approaches based on serology and also for vaccine formulation. The in- house ELISA is a promising alternative compared to invasive techniques.

Fatty acid methyl ester profiles and nutritive values of 20 marine microalgae in Korea

Objecive:To screen the fatty acid(FA) composition of 20 marine microalgae species,including seven Diophyceae,six Bacillariophyeae four Chlorophyceae,two Haptophyceae and one Raphidophyceae species.Methods:Microalgal cells cultured at the Korea Institute of Ocean Science & Technology were harvested during the late exponential growth phase and the FA composition analyzed.Results:The FA composition of microalgae was speciesspecific.For example,seven different species of Dinophyceae were composed primarily of C14:0,C16:0.C18:0.C20:4n-6.C20:5n-3 and C22:6n-3.while C14:0.C16:0,C16:1.C18:0.C20:5n-3 and C22:6n-3 were abundant FAs in six species of Bacillariophyceae.In addition,four Chlurophyceae,two Haptopkyeeae and one Raphidophyceae species all contained a high degree of C16:1 n-7[(9.2R-34.91)%and(34.48-35.04)%].C14:0[(13.34-25.96)%]and[(26.69-Z8.24)%],and C16:0[(5.89-29.15)%]and[(5.70-16.81)%].Several factors contribute to the nutritional value of microalgae.including the polyunsaturated FA content and n-3 to n-6 FA ratio,which could be used to assess the nutritional quality of microalgae.Conclusions:This study is the first comprehensive assessment of the FA composition and nutritional value of microalgae species in South Korea,and identifies the potential utility of FAs as species-specific biomarkers.

Atmospheric pressure gliding arc discharge plasma treatments for improving germination, growth and yield of wheat

Wheat （Triticum aestivum） seeds were treated with atmospheric pressure gliding arc discharge plasmas to investigate the effects on water absorption, seed germination rate, seedling growth and yield in wheat. The surface architectures and functionalities of the seeds were found to modify due to plasma treatments. 6 rain treatment was provided 95%-100% germination rate. For the treatment duration of 3 and 9 rain the growth activity, dry matter accumulation, leaves chlorophyll contents, longest spikes, number of spikes/spikelet and total soluble protein content in shoots were improved. The grain yield of wheat was increased ,--20% by 6 min treatment with H2O/O2 plasma with respect to control.

Detection of coat protein gene of nervous necrosis virus using loopmediated isothermal amplification

Objective:To establish a novel and highly specific loop-mediated isothermal amplification(LAMP) assay for the identification of nervous necrosis virus(NNV) infection.Methods:A set of synthesized primers was used to match the sequences of a specific region of the nnr gene from the National Center for Biotechnology Information database,not originating from NNV-infected fish,the efficiency and specificity of LAMP were measured dependent on the concentration of DNA polymerase and the reaction temperature and time.In addition,to determine species-specific LAMP primers,cross reactivity testing was applied to the reaction between NVV and other virus families including viral hemorrhagic septicemia virus and marine birnavirus.Results:The optimized LAMP reaction carried out at 64 ℃ for 60 min,and above 4 U Bst DNA polymerase.The sensitivity of LAMP for the detection of nnv was thus about 10 times greater than the sensitivity of polymerase chain reaction.The LAMP assay primers were specific for the detection NNV infection in Epinephelus septemfasciatus.Conclusions:The development of LAMP primers based on genetic information from a public database,not virus-infected samples,may provide a very simple and convenient method to identify viral infection in aquatic organisms.

Emblica officinalis improves glycemic status and oxidative stress in STZ induced type 2 diabetic model rats

Objective:To evaluate the antidiabetic and antioxidant potential of Emblica officinalis(E.officinalis)fruit on normal and type 2 diabetic rats.Methods:Type 2 diabetes was induced into the male Long-Evans rats.The rats were divided into nine groups including control groups receiving water,type 2 diabetic controls,type 2 diabetic rats treated with glibenclamide(T2GT)and type 2diabetic rats treated with aqueous extract of fruit pulp of E.officinalis.They were fed orally for8 weeks with a single feeding.Blood was collected by cutting the tail tip on 0 and 28 days and by decapitation on 56 day.Packed red blood cells and serum were used for evaluating different biochemical parameters.Results:Four weeks administration of aqueous extract of E.officinalis improved oral glucose tolerance in type 2 rats and after 8 weeks it caused significant(P<0.007)reduction in fasting serum glucose level compared to 0 day.Triglycerides decreased by 14%but there was no significant change in serum ALT,creatinine,cholesterol and insulin level in any group.Furthermore,reduced erythrocyte malondialdehyde level showed no significant change(P<0.07)but reduced glutathione content was found to be increased significantly(P<0.05).Conclusions:The aqueous extract of E.officinalis has a promising antidiabetic and antioxidant properties and may be considered for further clinical studies in drug development.

Characterization, prevalence and antibiogram study of Staphylococcus aureus in poultry

Objective: To reveal the presence of methicillin resistant Staphylococcus aureus(S. aureus)(MRSA) in poultry samples and to determine the antibiogram pattern against five antibiotics.Methods: Samples from different poultry farm of Chittagong city, Bangladesh were examined for S. aureus by different biochemical tests and confirmed as MRSA by identifying the presence of mec A gene using PCR. Antibiotic resistance pattern in S. aureus was determined by antibiotic disk diffusion method.Results: In this study, a total of 60 samples(30 from nasal swabs and 30 from cloacal swabs) were used, of which 54 were confirmed as S. aureus by different biochemical tests. Among these, 12 were confirmed as MRSA by detecting mec A gene using PCR.During antibiogram study, both nasal and cloacal samples showed the highest resistance against penicillin-G and the lowest resistance was observed against neomycin.Conclusions: Based on the present study, it can be said that different antibiotics are used extensively in poultry that leads to MRSA and is alarming for human health.

Construction of EST-SSR Databases for Effective Cultivar Identification and Their Applicability to Complement for Lettuce (Lactuca sativa L.) Distinctness Test

The objectives of this study were to construct a database of expressed sequence tag (EST)-simple sequence repeat (SSR) markers to identify lettuce cultivars. A set of 370 EST-SSR primer pairs were applied for fingerprinting the lettuce cultivars. Fifty-eight EST-SSR markers showed hy-per-variability and were able to differentiate 92 cultivars. A total of 176 polymorphic amplified fragments were obtained by the 58 markers, and two to eight SSR alleles were detected for each l&degCus with an average of three alleles per locus. Average polymorphism information content (PIC) was 0.425, ranging from 0.022 to 0.743. Cluster analysis was based on Jaccard’s distance coefficients using the method of unweighted pair group. In this method we used arithmetical averages (UPGMA) algorithm categorized 4 major groups, which were in accordance to morphological traits. The eight cultivars of three groups with 100% genetic similarity through SSR analysis were investigated by phenotypic traits. These cultivars including these pairs are very similar in 27 morphological characteristics. Therefore, these EST-SSR markers could be used to select similar cultivars through management of reference collection to complement distinctiveness test of lettuce cultivars.

Molecular characterization of Cryptosporidium xiaoi in goat kids in Bangladesh by nested PCR amplification of 18S rRNA gene

Objective:To investigate the prevalence of Cryptosporidium spp.in goat kids in selected areas of Bangladesh and to elucidate the potential zoonotic hazards.Methods:In the present study,we have used Ziehl-Neelsen staining and nested PCR approach to identify and characterize the Cryptosporidium sp.from diarrhoeic feces of goat kids.A total of 100 diarrhoeic feces samples were collected from Chittagong region in Southern Bangladesh.For nested PCR analysis,specific primers for amplification of 581 base pair fragments of 18 S rRNA gene were used.Results:A total of 15%and 3%samples were found positive in microscopic study and in nested PCR analysis respectively.Phylogenetic analysis of sequence data showed similarity with that of Cryptosporidium xiaoi recorded from sheep and goat.Conclusions:To our knowledge,this is the first report of Cryptosporidium xiaoi responsible for diarrhoea in goat kids in Bangladesh.Further study can highlight their zoonotic significance along with genetic diversity in other host species inside the country.

Protective Effect of Coleus aromaticus Benth(Lamiaceae)against Naphthalene-induced Hepatotoxicity

Objective To investigate protective effect of Coleus aromaticus leaf extract against naphthalene induced hepatotoxicity in rats. Methods Eighteen male rats were divided into three groups. Group I rats were treated as control. Group II rats were intraperitoneally administered with naphthalene （435 mg/kg b.wt） dissolved in corn oil once a day for a period of 30 days. Group III rats were treated with leaf extract （100 mg/kg b.wt） dissolved in 0.9% saline and naphthalene （435 mg/kg b.wt） dissolved in corn oil once a day for a period of 30 days. Results Significant protective effect was observed against naphthalene induced liver damage, which appeared evident from the response levels of marker enzymes （aspartate transaminase, alanine transaminase, acid phosphatase, alkaline phosphatase and lactate dehydrogenase）. The biochemical components viz. triglycerides, free fatty acids, cholesterol acyl transferase, high-density lipoprotein, low-density lipoprotein, cholesterol and bilirubin were found to be increased in liver and serum of naphthalene stressed rats when compared to control. Conclusion Treatment of naphthalene intoxicated rats with plant extract reversed these distorted parameters to near normal levels. Liver histology showed supportive evidence regarding the protective nature of plant extract against fatty changes induced by naphthalene. The present study provides a scientific rationale for using C. aromaticus in the management of liver disorders.

Biological and Functional Properties of Wedelolactone in Human Chronic Diseases

Medicinal herbs are well known and studied over the past millennia in most of the developing countries as a rational means of treatment against various diseases and disorders.Wedelolactone(WDL),a major bioactive compound in Eclipta prostrata L(Eclipta alba L),has been reported with potential benefits in human health against chronic diseases.However,a comprehensive study on WDL pharmacological benefits in various ailments,to the best of our knowledge,is not yet reported.Thereof,the present review provides the recent therapeutic applications in reference to biological and functional activities against major human chronic diseases,including cardiovascular,cancer,diabetes mellitus,liver disease,Alzheimer’s disease,and androgenetic alopecia.In this study,we collected all the relevant experimental information on WDL from Scientific databases such as PubMed,Web of Science,Science Direct,and Google Scholar.Conclusively,WDL is recognized as a key anti-oxidant with both specific regulator and inhibitor of major drug targetable proteins in human chronic diseases and disorders.Hence,WDL as a novel therapeutic bioactive molecule is advised to explore further for relevant pharmacological activities.

Effects of zinc and nitrogen fertilizer and their application method on yield and yield components of <i>Phaseolus vulgaris</i>L.

An experiment in factorial format based on randomized complete block design with 3 replications was conducted to study the effects of zinc spray (0 and 1 g/L) and nitrogen fertilizer (0, 25, 50 and 75 kg/ha pure nitrogen) on yield and yield components of Phaseolus vulgaris. In maturity time, seed yield, 100 seed weight, number of pods per plant, number of seeds per pod and plant height were measured. Results showed that, use of zinc spray had a significant effect in 1% probability level on all measured traits. Also, the effect of nitrogen on all studied traits was significant in 1% probability level. Interaction effect of zinc spray and nitrogen fertilizer on number of seed per pod in 1% and on seed yield and plant height in 5% was significant and on other traits was non significant. The highest seed yield was obtained by zinc spray application with 1996 kg/ha. Among nitrogen fertilizer levels, use of 90 kg/ha pure nitrogen showed highest seed yield.

Efficient detection of pathogen virus in sand dabs,Paralichthys olivaceus using loop-mediated isothermal amplification(LAMP)

Viral hemorrhagic septicemia virus（VHSV） and marine birnavirus（MABV） are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogen infection. The loop-mediated isothermal amplification（LAMP） of DNA is currently one of the most commonly used molecular diagnostic tools, as it is simple, quick, and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fish was developed. Using a set of synthesized primers matching a specific region of the genome, the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficient detection of VHSV and MABV infection in the wild fish, Paralichthys olivaceus. Our results demonstrate the simplicity and convenience of this method for the detection of viral infection in aquatic organisms.

In silico identification and characterization of common epitope-based peptide vaccine for Nipah and Hendra viruses

Objective: To explore a common B-and T-cell epitope-based vaccine that can elicit an immune response against encephalitis causing genus Henipaviruses, Hendra virus(He V) and Nipah virus(Ni V). Methods: Membrane proteins F, G and M of He V and Ni V were retrieved from the protein database and subjected to different bioinformatics tools to predict antigenic B-cell epitopes. Best B-cell epitopes were then analyzed to predict their T-cell antigenic potentiality. Antigenic B-and T-cell epitopes that shared maximum identity with He V and Ni V were selected. Stability of the selected epitopes was predicted. Finally, the selected epitopes were subjected to molecular docking simulation with HLA-DR to confirm their antigenic potentiality in silico. Results: One epitope from G proteins, one from M proteins and none from F proteins were selected based on their antigenic potentiality. The epitope from the G proteins was stable whereas that from M was unstable. The M-epitope was made stable by adding flanking dipeptides. The 15-mer G-epitope(VDPLRVQWRNNSVIS) showed at least 66% identity with all Ni V and He V G protein sequences, while the 15-mer M-epitope(GKLEFRRNNAIAFKG) with the dipeptide flanking residues showed 73% identity with all Ni V and He V M protein sequences available in the database. Molecular docking simulation with most frequent MHC class-II(MHC II) and class-I(MHC I) molecules showed that these epitopes could bind within HLA binding grooves to elicit an immune response. Conclusions: Data in our present study revealed the notion that the epitopes from G and M proteins might be the target for peptide-based subunit vaccine design against He V and Ni V. However, the biochemical analysis is necessary to experimentally validate the interaction of epitopes individually with the MHC molecules through elucidation of immunity induction.

Investigation of in vivo neuropharmacological effect of Alpinia nigra leaf extract

Objective:To analyze in vivo neuro-pharmacological effects of Alpinia nigra as anxiety is a particular form of behavioral inhibition that occurs in response to novel environmental events.Methods:In present study,the extract of Alpinia nigra was evaluated for its central nervous system depressant effect using mice behavioral models,such as hole cross,open field and thiopental sodium induced sleeping time tests for its sedative properties and an elevated plusmaze test for its anxiolytic potential,respectively.Results:In anxiolytic study,the extract displayed increased percentage of entry into open arm ut the dose of 400 and 200 mg/kg.The extract produced a significant(P<0.01) increase in sleeping duration and reduction of onset of sleep compared to sodium thiopental at both doses(200 and400 mg/kg).The extract t.200 and 400 mg/kg) also showed a dose-dependent suppression of motor activity and exploratory activity of the mice in both open field and hole cross test.Conclusion:This study demonstrates that the treated extract has significant central nervous system depressant effect.Further studies on active constituent of the extract can provide approaches for therapeutic intervention.