Asian Pacific Journal of Tropical Biomedicine

89-94

Authors: Freddie Masaninga, Emmanuel Chanda, Pascalina Chanda-Kapata, Busiku Hamainza, Hieronymo T Masendu, Mulakwa Kamuliwo, Wambinji Kapelwa, John Chimumbwa, John Govere, Mac Otten, Ibrahima Soce Fall, Olusegun Babaniyi

A comprehensive desk review of malaria trends was conducted between 2000-2010 in Zambia to study malaria epidemiology and trends to guide strategies and approaches for effective malaria control. This review considered data from the National Health Information Management System, Malaria Surveys and Programme Review reports and analyzed malaria in-patient cases and deaths in relation to intervention coverage for all ages. Data showed three distinct epidemiological strata after a notable malaria reduction (66%) in in-patient cases and deaths, particularly between 2000-2008. These changes occurred following the (re-)introduction and expansion of indoor residual spraying up to 90% coverage, scale-up of coverage of long-lasting insecticidetreated nets in household from 50% to 70%, and artemisin-based combination therapy nationwide. However, malaria cases and deaths re-surged, increasing in 2009-2010 in the northern-eastern parts of Zambia. Delays in the disbursement of funds affected the implementation of interventions, which resulted in resurgence of cases and deaths. In spite of a decline in malaria disease burden over the past decade in Zambia, a reversal in impact is notable in the year 2009-2010, signifying that control gains are fragile and must be sustained toeliminate malaria.

100-104

Authors: Wabo Poné J, Payne V K, Mbogning Tayo Gertrude, Komtangi Marie Claire, Yondo Jeannette, Ngangout Alidou M, Mpoame Mbida, Bilong Bilong CF

Objective: To evaluate the ovicidal and larvicidal activities of aqueous and ethanolic extracts of leaves of Dichrocephala integrifolia (D. integrifolia) against the eggs (fresh and embryonnated), the first and second larval stages of Heligmosomoides bakeri. In order to verify if this medicinal plant possesses active compounds capable of inhibiting the embryonation and hatching of eggs or to induce the mortality of larvae (L1 and L2). Methods: Dried extracts were diluted in distilled water to obtain five different concentrations: 625, 1250, 2500, 3750 and 5000 μg/mL. Fresh eggs obtained from artificially infected mice feces were exposed to these different concentrations for 48 h. Time of contact for embryonated eggs was 6 h while L1 and L2 larvae were exposed for 24 h. Distilled water (placebo) and 1.5% DMSO were used as negative controls. Results: Distilled water, and 1.5% DMSO had no effect on embryonation, hatching and larval survival. Aqueous extracts of D. integrifolia showed a weak activity against all stages of the parasite at all concentrations tested. On the contrary, the ethanolic extract of D. integrifolia inhibited the embryonation of 87.5% of fresh eggs, the hatching of 81.1% of embryonated eggs and induced the mortality of 98.1% and 98% of L1 and L2 larvae respectively at 5 000 μg/mL. Conclusions: The results of the present study indicate that the ethanolic extracts of D. integrifolia contained compounds with ovicidal and larvicidal properties. In spite of these results, in vivo tests, studies on toxicity and mechanism of action of active compounds are also needed to validate the utilisation of this medicinal plant by population of Dschang-Cameroon to treat gastro-intestinal parasites.

111-117

Authors: Zulfaidah Penata Gama, Nobukazu Nakagoshi, Suharjono, Faridah Setyowati

Objective To investigate the toxicity of indigenous Bacillus thuringiensis (B. thuringiensis)isolates from Malang City for controlling Aedes aegypti (Ae. aegypti) larvae. Methods Soil samples were taken from Purwantoro and Sawojajar sub-districts. Bacterial isolation was performed using B. thuringiensis selective media. Phenotypic characteristics of the isolates were obtained with the simple matching method. The growth and prevalence of spores were determined by the Total Plate Count method, and toxicity tests were also performed on the third instar larval stage of Ae. aegypti. The percentage of larval mortality was analysed using probit regression. The LC50 was analysed by ANOVA, and the Tukey HSD interval was 95%. Results Among the 33 selected bacterial isolates, six were obtained (PWR4-31, PWR4-32, SWJ4-2b, SWJ4-4b, SWJ-4k and SWJ5-1) that had a similar phenotype to reference B. thuringiensis. Based on the dendrogram, all of the bacterial isolates were 71% similar. Three isolates that had a higher prevalence of reference B. thuringiensis were PWR4-32, SWJ4-4b and SW5-1, of which the spore prevalence was 52.44%, 23.59%, 34.46%, respectively. These three indigenous isolates from Malang City successfully killed Ae. aegypti larvae. The PWR4-32 isolates were the most effective at killing the larvae. Conclusions Six indigenous B. thuringiensis isolates among the 33 bacterial isolates found in the Sawojajar and Purwantoro sub-districts were toxic to the third instar larvae of Ae. aegypti. The PWR4-32 isolates were identical to the reference B. thuringiensis and had 88% phenotype similarity. The PWR4-32 isolates had the highest spore prevalence (52.44%), and the early stationary phase occurred at 36 h. The PWR4-32 isolates were the most effective at killing Ae. aegypti larvae (LC50-72 h=2.3×108 cells/mL).

123-128

Authors: Abdollah Ghasemi Pirbalouti, Maryam Mohammadi

Objective To examine the chemical variability in inflorescences of wild populations of Stachys lavandulifolia Vahl (S. lavandulifolia) collected throughout two provinces (Isfahan and Chaharmahal va Bakhtiary), Southwest Iran. Methods The essential oils of S. lavandulifolia Vahl from seven locations were obtained by hydro-distillation and analysed by gas chromatography and gas chromatography-mass spectrometry. Results The results revealed that distinct differences in the content of compounds depending on region of sample collection. The main constituents of the essential oils were α-thujone (0.3%-32.3%), α-pinene (trace to 37.3%), myrcene (0.5%-15.9%), β-phellandrene (1.1%-37.9%), germacrene D (0.4%-11.3%), Δ-cadinene (trace to 11.6%) and 1, 4-methano-1 H-indene (trace to 10.1%). Conclusions The results of the present study indicated that essential oil components of S. lavandulifolia Vahl can be varied with genetic (ecotype), environmental conditions and geographic origin. In general, the essential oils of various populations of S. lavandulifolia Vahl were rich in monoterpenoids and sesquiterpenoids.

140-145

Authors: Reza Mirnejad, Sepideh Mostofi, Faramaz Masjedian

Objective To investigate antibiotic resistance and carriage class 1 and 2 integrons in clinical isolates of Acinetobacter baumannii (A. baumannii) from Tehran, Iran. Methods Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute. The presence of integrons was investigated by PCR using specific primers. Results Among isolated A. baumannii strains, 82% were multidrug resistant, 27 samples (54%) were resistant to three or more than three antibiotics and 16 samples (32%) showed resistance to two antibiotics. Integrons were detected from 44 of 50 isolates (88%), with classes 1 and 2 being observed in 42% (21/50) and 82% (41/50) of isolates, respectively. Integron-positive A. baumannii isolates showed higher antibiotic resistance than integron-negative isolates and all showed a multidrug-resistant phenotype. Conclusions Our findings show that classes 1 and 2 integrons, and especially classes 2 integrons are widely disseminated among A. baumannii strains isolated from Tehran and these structures are playing a major role in the acquisition of multidrug resistance in these strains. So monitoring of drug resistance with investigating carriage class 1 and 2 integrons is very important to plan specific infection control measures due to multidrug resistance A. baumannii in Iran hospitals.

148-153

Authors: Debasmita Dubey, Shakti Rath, Mahesh C. Sahu, Subhrajita Rout, Nagen K. Debata, Rabindra N. Padhy

Objective To investigate the infection of hospital- and community-acquired “erythromycin-induced clindamycin resistant” strains or D-test positives of clinical isolates of Staphylococcus aureus (S. aureus) (with and without methicillin resistance) in a hospital. Methods Strains of S. aureus isolated from clinical specimens were subjected to D-test and antibiotic profiling. Results Of the total 278 isolates, 140 (50.35%) were D-test positives and the rest were D-test negatives. Further, of 140 (100%) positives, 87 (62.14%) and 53 (37.85%) strains were from males and females, respectively. Of 140 (100%) positives, 117 (83.57%) were methicillin resistant S. aureus and 23 (16.42%) were methicillin sensitive S. aureus; of 140 strains, 103 (73.57%) strains from persons with and 37 (26.42%) were without related infections; of 140 strains, 91 (65%) and 49 (35%) were from hospital- and community-acquired samples, respectively. In 140 strains, 118 (84.28%) with comorbidities and 22 (15.71%) without comorbidities cases were recorded; similarly, persons with prior antibiotic uses contributed 108 (77.14%) and without 32 (22.85%) positive strains. These binary data of surveillance were analyzed by a univariate analysis. It was evident that the prior antibiotic uses and comorbidities due to other ailments were the determinative factors in D-test positivity, corroborated by low P values, P=0.001 1 and 0.002 4, respectively. All isolates (278) were resistant to 17 antibiotics of nine groups, in varying degrees; the minimum of 28% resistance for vancomycin and the maximum of 97% resistance for gentamicin were recorded. Further, of 278 strains, only 42 (15.1%) strains were resistant constitutively to both antibiotics, erythromycin resistant and clindamycin resistant, while 45 (16.2%) strains were constitutively sensitive to both antibiotics (erythromycin sensitive and clindamycin sensitive). Further, of the rest 191 (68.7%) strains were with erythromycin resistant and clindamycin resistant, of which only 140 (50.35%) strains were D-test positives, while the rest 51 (18.34%) strains were D-test negatives. Conclusions In view of high prevalence of D-test positive S. aureus strains, and equally high prevalence of multidrug resistant strains both in community and hospital sectors, undertaking of D-test may be routinely conducted for suppurative infections.

156-162

Authors: Vivek Kumar Vyas, Keyur Brahmbhatt, Hardik Bhatt, Utsav Parmar

Many active secretions produced by animals have been employed in the development of new drugs to treat diseases such as hypertension and cancer. Snake venom toxins contributed significantly to the treatment of many medical conditions. There are many published studies describing and elucidating the anti-cancer potential of snake venom. Cancer therapy is one of the main areas for the use of protein peptides and enzymes originating from animals of different species. Some of these proteins or peptides and enzymes from snake venom when isolated and evaluated may bind specifically to cancer cell membranes, affecting the migration and proliferation of these cells. Some of substances found in the snake venom present a great potential as anti-tumor agent. In this review, we presented the main results of recent years of research involving the active compounds of snake venom that have anticancer activity.

8-16

Authors: Kajal Chakraborty, Nammunayathuputhenkotta Krishnankartha Praveen, Kodayan Kizekadath Vijayan, Gonugontla Syda Rao

Objective: To evaluate the antioxidant activities and total phenolic contents of brown seaweeds belonging to Turbinaria spp. [Turbinaria conoides (T. conoides) and Turbinaria ornata (T. ornata)collected from Gulf of Mannar of southeastern coast of India in various in vitro systems. Methods:The antioxidant activity was evaluated using different in vitro systems, viz., 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2, 2′-azino-bis-3ethylbenzothiozoline-6-sulfonic acid diammonium salt(ABTS), H2O2/HO radical scavenging, Fe2+ ion chelating ability, and reducing potential. FolinCiocalteu method was used to determine the total phenolic content of the extracts, and the results were expressed as mg of gallic acid equivalents (GE)/g of the seaweed extracts. Thiobarbituric acid-reactive substances assay was employed to assess the ability of the seaweed extracts to inhibit lipid oxidation. Results: Ethyl acetate (EtOAc) fraction of T. conoides registered significantly higher phenolic content (105.97 mg GE/g) than that of T. ornata (69.63 mg GE/g). Significantly higher antioxidant potential as determined by DPPH (64.14%) radical scavenging acti vity was registered in EtOAc fraction of T. ornata. A higher ABTS.+ radical scavenging (IC50 3.16 µg/mL),Fe2+ chelating (IC50 0.46 mg/mL), H2O2 scavenging (IC50 4.25 mg/mL), lipid peroxidation inhibitory (TBARS, IC50 0.21 mg/mL), and reducing abilities (IC50 52.67 mg/mL) (P<0.05) were realized in EtOAc fraction of T. ornata than other fractions. Conclusions: This study indicated the potential use of T. conoides and T. ornata as candidate species to be used as food supplements/functional foods to increase shelf-life of food items for human consumption, and nutraceuticals to deter deleterious free radical-induced life-threatening diseases.

22-27

Authors: Shahrokh Bagheri, Hassan Ahmadvand, Ali Khosrowbeygi, Farshid Ghazanfari, Narges Jafari, Habibolah Nazem, Reza Haji Hosseini

Objective: To assess various antioxidative activities of Satureja khozestanica essential oil(SKE) and its effect on oxidation of low density lipoprotein (LDL) induced by CuSO4 in vitro by monitoring the formation of conjugated dienes and malondialdehyde (MDA). Methods: The formation of conjugated dienes, lag time and MDA were measured. Inhibition of this Cu-induced oxidation was studied in the presence of several concentrations of SKE. Also total antioxidant activity and free radical scavenging of SKE were determinated. Results: It was demonstrated that SKE was able to inhibit LDL oxidation and decrease the resistance of LDL against oxidation. The inhibitory effects of SKE on LDL oxidation were dose-dependent at concentrations ranging from 50 to 200 μg/mL. Total antioxidant capacity of SKE was (3.20±0.40) nmol of ascorbicacid equivalents/g SKE. The SKE showed remarkable scavenging activity on 2, 2-diphenylpicrylhydrazyl, IC50 (5.30± 0.11) ng/mL. Conclusions: This study shows that SKE is a source of potent antioxidants and prevents the oxidation of LDL in vitro and it may be suitable for use in food and pharmaceutical applications.

35-40

Authors: Nabila Benariba, Rabeh Djaziri, Wafaa Bellakhdar, Nacera Belkacem, Marcel Kadiata, Willy J. Malaisse, Abdullah Sener

Objective: To study the phytochemical screening of different extracts from Citrullus colocynthis (C. colocynthis ) seeds extracts and to assess their antioxidant activity on the DPPH free radical scavenging. Methods: Phytochemical screening, total content of polyphenols and flavonoids of C. colocynthis seeds extracts, including a crude aqueous extract (E1), a defatted aqueous extract (E2), a hydromethanolic extract (HM), an ethyl acetate extract (EA) and a n-butanol extract (n-B) was carried out according to the standard methods and to assess their corresponding effecton the antioxidant activity of this plant. Results: None of these extracts contained detectable amount of alkaloid, quinone, antraquinone, or reducing sugar. Catechic tannins and flavonoids were abundant in E1, HM and EA, whilst terpenoids were abundantly present in E1 and n-B but only weekly in HM. Coumarins were found in E2, EA and n-B. Polyphenols, expressed as gallic acid equivalent, amounted, per 100 g plant matter, to 329, 1002 and 150 mg in EA, HM an E1 respectively. Flavonoids, expressed as catechin equivalent, amounted, per 100 g plant matter to 620, 241 and 94 mg in EA, HM and E1 respectively. Comparable values were found in n-B and E1, with lower values in E2. Quercetin, myricetin and gallic acid were found in the EA and HM extracts by thin layer chromatography, The antioxidative effect of these extracts yielded, when tested at a concentration of 2 000 μg/mL in a 1,1-diphenyl-2-picrylhydrazyl assay, a reducing percentage of 88.8% with EA, 74.5% with HM and 66.2% with E1, and corresponding IC50 of 350, 580 and 500 μg/mL as compared to 1.1 μg/mL for ascorbic acid. Conclusions: These qualitative and quantitative analytical data document the presence in C. colocynthis extracts of such chemical compounds as flavonoids responsible for the antioxidant activity, as well as other biological activities of this plant.

47-52

Authors: Sirikamon Saengmee-anupharb, Toemsak Srikhirin, Boonyanit Thaweboon, Sroisiri Thaweboon,Taweechai Amornsakchai, Surachai Dechkunakorn, Theeralaksna Suddhasthira

Objective: To evaluate the antimicrobial activities of silver inorganic materials, including silver zeolite (AgZ), silver zirconium phosphate silicate (AgZrPSi) and silver zirconium phosphate (AgZrP), against oral microorganisms. In line with this objective, the morphology and structure of each type of silver based powders were also investigated. Methods: The antimicrobial activities of AgZ, AgZrPSi and AgZrP were tested against Streptococcus mutans, Lactobacillus casei, Candida albicans and Staphylococcus aureus using disk diffusion assay as a screening test. The minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were determined using the modified membrane method. Scanning electron microscope and X-ray diffraction were used to investigate the morphology and structure of these silver materials. Results: All forms of silver inorganic materials could inhibit the growth of all test microorganisms. The MIC of AgZ, AgZrPSi and AgZrP was 10.0 g/L whereas MLC ranged between 10.0-60.0 g/L. In terms of morphology and structure, AgZrPSi and AgZrP had smaller sized particles (1.5-3.0 µm) and more uniformly shaped than AgZ. Conclusions: Silver inorganic materials in the form of AgZ, AgZrPSi and AgZrP had antimicrobial effects against all test oral microorganisms and those activities may be influenced by the crystal structure of carriers. These results suggest that these silver materials may be useful metals applied to oral hygiene products to provide antimicrobial activity against oral infection.

58-63

Authors: Ghassan Mohammad Sulaiman, Wasnaa Hatif Mohammed, Thorria Radam Marzoog, Ahmed Abdul Amir AlAmiery, Abdul Amir H. Kadhum, Abu Bakar Mohamad

Objective: To synthesize silver nanopaticles from leaves extract of Eucalyptus chapmaniana (E. chapmaniana) and test the antimicrobial of the nanoparticles against different pathogenic bacteria, yeast and its toxicity against human acute promyelocytic leukemia (HL-60) cell line. Methods: Ten milliliter of leaves extract was mixed with 90 mL of 0.01 mmol/mL or 0.02 mmol/mL aqueous AgNO3 and exposed to sun light for 1 h. A change from yellowish to reddish brown color was observed. Characterization using UV-vis spectrophotometery and X-ray diffraction analysis were performed. Antimicrobial activity against six microorganisms was tested using well diffusion method and cytoxicity test using 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, a yellow tetrazole was obtained on the human leukemia cell line (HL-60). Results: UVvis spectral analysis showed silver surface plasmon resonance band at 413 nm. X-ray diffraction showed that the particles were crystalline in nature with face centered cubic structure of the bulk silver with broad beaks at 38.50 ° and 44.76 °. The synthesized silver nanoparticles efficiently inhibited various pathogenic organisms and reduced viability of the HL-60 cells in a dosedependent manner. Conclusions: It has been demonstrated that the extract of E. chapmaniana leaves are capable of producing silver nanoparticles extracellularly and the Ag nanoparticles are quite stable in solution. Further studies are needed to fully characterize the toxicity and the mechanisms involved with the antimicrobial and anticancer activity of these particles.

69-73

Authors: Laila Salim Al Hashmi, Mohammad Amzad Hossain, Afaf Mohammed Weli, Qasim Al-Riyami, Jamal Nasser AlSabahi

Objective: To isolate and analyze the chemical composition in different crude extracts of from the leaves of locally grown of Thymus vulgaris L (T. vulgaris) by gas chromatographymass spectrometry (GC-MS). Methods: The shade dried leaves powder was extracted with methanol by using Soxhlet extractor. Methanol crude extracts of T. vulgaris and the derived fractions of hexane, chloroform, ethyl acetate and butanol were obtained. Results: Qualitative analyses of various organic crude extracts of T. vulgaris by using GC-MS showed that there were different types of high and low molecular weight compounds. Most of the isolated and identified compounds by GC-MS in the crude extracts are basically biologically important. Further, the T.vulgaris leaf possessed certain characteristics that can be ascribed to cultivation on a domestic plantation. The crude extracts were prepared from the powder leaves of T. vulgaris for respective compounds can be chosen on the basis of above GC-MS analysis. Conclusions: All the major compounds were identified and characterized by spectroscopic method in different organic crude extracts of T. vulgaris are biologically active molecules. Thus the identification of a good number of compounds in various crude extracts of T. vulgaris might have some ecological role.

79-84

Authors: Rajamani Saranya, Thirunavukkarasu Thirumalai, Munisami Hemalatha, Ranganathan Balaji, Ernest David

Traditional medicine system of India comprised varieties of plants which are playing a significant role in curing diseases from ancient times. Among them, Enicostemma littorale blume (E.littorale) a perennial herb of the family Gentianaceae is cosmopolitan in occurrence in India. The bittery natured plant acts as a laxative, helps in curing fever, rheumatism, skin diseases, abdominal disorders, snake bite, obesity and helps to regulate blood sugar levels. The plant constituents have been reported for possessing antimicrobial, antioxidant, antiulcer, antiinflammatory, hypolipidaemic, hepatoprotective and hypoglycemic properties. This review provides a bird’s eye view about geographical distribution, physicochemical parameters, phytoconstituents and pharmacological properties of E. littorale.

930–935

Authors: P Kumar, S Senthamil Selvi, M Govindaraju

Objective To investigate the anti-biofilm and anti-bacterial activity of Junceella juncea (J. juncea) against biofilm forming pathogenic strains. Methods Gorgonians were extracted with methanol and analysed with fourier transform infrared spectroscopy. Biofilm forming pathogens were identified by Congo red agar supplemented with sucrose. A quantitative spectrophotometric method was used to monitor in vitro biofilm reduction by microtitre plate assay. Anti-bacterial activity of methanolic gorgonian extract (MGE) was carried out by disc diffusion method followed by calculating the percentage of increase with crude methanol (CM). Results The presence of active functional group was exemplified by FT-IR spectroscopy. Dry, black, crystalline colonies confirm the production of extracellular polymeric substances responsible for biofilm formation in Congo red agar. MGE exhibited potential anti-biofilm activity against all tested bacterial strains. The anti-bacterial activity of methanolic extract was comparably higher in Salmonella typhii followed by Escherichia coli, Vibrio cholerae and Shigella flexneri. The overall percentage of increase was higher by 50.2% to CM. Conclusions To conclude, anti-biofilm and anti-bacterial efficacy of J. juncea is impressive over biofilm producing pathogens and are good source for novel anti-bacterial compounds.

944–947

Authors: Rabia Nazm, Asghari Bano

Objective To investigate the in vitro antimicrobial activities of the leaf extract in different solvents viz., methanol, ethanol and water extracts of the selected plant Ricinus communis. Methods Agar well diffusion method and agar tube dilution method were carried out to perform the antibacterial and antifungal activity of methanol, ethanol and aqueous extracts. Results Methanol leaf extracts were found to be more active against Gram positive bacteria (Bacillus subtilis: ATCC 6059 and Staphylococcus aureus: ATCC 6538) as well as Gram negative bacteria (Pseudomonas aeruginosa: ATCC 7221 and Klebsiella pneumoniae) than ethanol and aqueous leaf extracts. Antifungal activity of methanol and aqueous leaf extracts were also carried out against selected fungal strains as Aspergillus fumigatus and Aspergillus flavus. Methanolic as well as aqueous leaf extracts of Ricinus communis were effective in inhibiting the fungal growth. Conclusions The efficient antibacterial and antifungal activity of Ricinus communis from the present investigation revealed that the methanol leaf extracts of the selected plant have significant potential to inhibit the growth of pathogenic bacterial and fungal strains than ethanol and aqueous leaf extracts.

953–959

Authors: Swetha Sunkar, C Valli Nachiyar

Objective To synthesize the ecofriendly nanoparticles, which is viewed as an alternative to the chemical method which initiated the use of microbes like bacteria and fungi in their synthesis. Methods The current study uses the endophytic bacterium Bacillus cereus isolated from the Garcinia xanthochymus to synthesize the silver nanoparticles (AgNPs). The AgNPs were synthesized by reduction of silver nitrate solution by the endophytic bacterium after incubation for 3-5 d at room temperature. The synthesis was initially observed by colour change from pale white to brown which was confirmed by UV-Vis spectroscopy. The AgNPs were further characterized using FTIR, SEM-EDX and TEM analyses. Results The synthesized nanoparticles were found to be spherical with the size in the range of 20-40 nm which showed a slight aggregation. The energy-dispersive spectra of the nanoparticle dispersion confirmed the presence of elemental silver. The AgNPs were found to have antibacterial activity against a few pathogenic bacteria like Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Klebsiella pneumoniae. Conclusions The endophytic bacteria identified as Bacillus cereus was able to synthesize silver nanoparticles with potential antibacterial activity.

966–974

Authors: Sivagnanam Selva Kumar, Mudiganti Ram Krishna Rao, Maruthaiveeran Periyasamy Balasubramanian

Objective To find out the anticancer effect of Indigofera aspalathoides (I. aspalathoides) on 20-methylcholanthrene induced fibrosarcoma in rats. Methods Fibrosarcoma was induced in Wistar strain male albino rats by 20-methylcholanthrene. Intraperitoneous (i.p.) administration of 250 mg/kg body weight/day of aqueous extract of I. aspalathoides for 30 d effectively suppressed chemically induced tumors. Parameters such as body weight, liver and kidney weight, tumor weight, mean survival time, behavioral changes, blood glucose, blood glycogen and marker enzymes such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), acid phosphatase (ACP) and 5′-nucleiotidase (5′-NT) in serum, liver and kidney and lipid profiles such as total cholesterol, phospholipids, free fatty acids in liver and kidney of control and experimental animals were studied. Results Fibrosarcoma bearing animals were ferocious and anxious. The mean survival time was found to increase after the treatment. The body weights were significantly decreased (P<0.001) in group II fibrosarcoma animals which steadily increased after the treatment with I. aspalathoides. The liver and kidney weights were significantly increased whereas the tumor weights decreased as compared to the weights in untreated fibrosarcoma bearing rats. The blood glucose and the liver and kidney glycogen levels were found to decrease significantly (P<0.001) in group II animals. Elevated activities of marker enzymes were observed in serum, liver and kidney of fibrosarcoma bearing Group II animals which were normalize after I. aspalathoides treatment. In the liver and kidney of Group II animals the total cholesterol increased whereas the phospholipids and free fatty acid levels decreased (P<0.001) which were normalized after treatment. Conclusions The treatment by I. aspalathoides on fibrosarcoma bearing rats has improved the levels of various parameters indicating its antiproliferative and anticancer activity.

981–986

Authors: Alagarmalai Jeyasankar, Selvaraj Premalatha, Kuppusamy Elumalai

Objective To evaluate the antifeedant, insecticidal and growth inhibition activities of Solanum pseudocapsicum (S. pseudocapsicum) seed extracts against Spodoptera litura (S. litura) and Helicoverpa armigera (H. armigera). Methods Hexane, diethyl ether, dichloromethane and ethyl acetate seed extracts were prepared and tested for antifeedant, insecticidal and growth inhibitory activities against fourth instar larvae of S. litura and H. armigera. Results Ethyl acetate extract showed promising antifeedant and insecticidal activities against S. litura and H. armigera. Percentage of deformed larvae, pupae and adults were maximum in treatment of ethyl acetate extract. Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae. Conclusions Ethyl acetate extracts of S. pseudocapsicum, showed higher efficiency of antifeedant, insecticidal and growth inhibition activities. Hence, it can be used to controll agricultural insect pests, S. litura and H. armigera.

992–998

Authors: Urmila M Aswar, Padmaja P Kalshetti, Suhas M Shelke, Sharad H Bhosale, Subhash L Bodhankar

Objective To study the derivatives of 1,2,4-triazino[5,6-b]indole-3-thione for antidepressant activity in olfactory bulbectomized (OBX) rats. Out of various derivatives tested for acute tail suspension test, the two derivatives showing prominent action were selected for bilateral olfactory bulbectomy model of chronic depression in rats. Methods The sub acute effects of 14-day oral pretreatment of two derivatives labeled as 3a (70 mg/kg) and 3r (70 mg/kg), imipramine (20 mg/kg), fluoxetine (30 mg/kg) and moclobemide (15 mg/kg) were evaluated on bilateral bulbectomy induced rise in body weight, hyperphagia, hyperactivity, and on sexual dysfunction. The serum sodium concentration, body temperature, and heart rate were also recorded. Results The derivatives 3a and 3r showed reversal of drop in body weight, reversed OBX induced hyperactivity, normalized body temperature, heart rate, and serum sodium concentration. In elevated maze test, moclobemide, 3a, 3r treatment significantly reduced time spent in open arm as compared to OBX rats. 3a and 3r also improved sexual behavior parameters. Conclusions The present study shows promising antidepressant action and provides a proof of concept for the chronic treatment of 3a, 3r to treat depression.

1002–1008

Authors: Priyanka Soni, Anees Ahmad Siddiqui, Jaya Dwivedi, Vishal Soni

India has a great wealth of various naturally occurring plant drugs which have great potential pharmacological activities. Datura stramonium (D. stramonium) is one of the widely well known folklore medicinal herbs. The troublesome weed, D. stramonium is a plant with both poisonous and medicinal properties and has been proven to have great pharmacological potential with a great utility and usage in folklore medicine. D. stromonium has been scientifically proven to contain alkaloids, tannins, carbohydrates and proteins. This plant has contributed various pharmacological actions in the scientific field of Indian systems of medicines like analgesic and antiasthmatic activities. The present paper presents an exclusive review work on the ethnomedical, phytochemical, pharmacological activities of this plant.

849–857

Authors: Al Thabiany Aziz, Hamady Dieng, Abu Hassan Ahmad, Jazem A Mahyoub, Abdulhafis M Turkistani, Hatabbi Mesed, Salah Koshike, Tomomitsu Satho, MR Che Salmah, Hamdan Ahmad, Wan Fatma Zuharah, Ahmad Saad Ramli, Fumio Miake

Objective To investigate the prevalence of container breeding mosquitoes with emphasis on the seasonality and larval habitats of Aedes aegypti (Ae. aegypti) in Makkah City, adjoining an environmental monitoring and dengue incidence. Methods Monthly visits were performed between April 2008 and March 2009 to randomly selected houses. During each visit, mosquito larvae were collected from indoors and outdoors containers by either dipping or pipetting. Mosquitoes were morphologically identified. Data on temperature, relative humidity, rain/precipitations during the survey period was retrieved from governmental sources and analyzed. Results The city was warmer in dry season (DS) than wet season (WS). No rain occurred at all during DS and even precipitations did fall, wetting events were much greater during WS. Larval survey revealed the co-breeding of Aedes, Culex and Anopheles in a variety of artificial containers in and around homes. 32 109 larvae representing 1st , 2nd, 3rd, and 4th stages were collected from 22 618 container habitats. Culicines was far the commonest and Aedes genus was as numerous as the Culex population. Ae. aegypti larval abundance exhibited marked temporal variations, overall, being usually more abundant during WS. Ten types of artificial containers were found with developing larvae. 70% of these habitats were located indoors. 71.42% of indoor containers were permanent and 28.58% was semi-permanent during WS. Cement tanks was the only container type permanent during DS. Ae. aegypti larval indices (CI, HI, BI) recorded were greater during WS. Conclusions Taken together, these results indicate a high risk of dengue transmission in the holy city.

863–869

Authors: Hala F Mohamed

Objective To characterize, identify and investigate the anticancer properties of two new soil fungal isolates, Emericella nidulans and Fusarium solani isolated from Wady El-Natron in Egypt against colon cancer Caco-2 (ATCC) cell line. Methods Soil sample was cultured and two strains were chosen for morphological and phenotypical characterization. Partial sequences of the 18s rRNA gene and the internal transcribed spacer region ITS of the two isolates were amplified by PCR. Phylogenetic tree construction and analysis of the resulted multiple sequences from the two fugal isolates were also carried out. In vitro anticancer activity of the two strains was done against colon Caco-2 cancer cell line. Reverse transcription – PCR was carried out to detect level of expression of p53 in Caco-2 cell line. Results HF.1 displayed morphological and genotypic characteristics most similar to that of Fusarium solani while HF.2 was most similar to Emericella nidulans with high similarity of 99% and 97% respectively. The multiple sequence alignment of the two fungal isolates showed that, the maximum identical conserved domains in the 18s rRNA genes were identified with the nucleotide regions of 51st to 399th base pairs, 88th to 525th base pairs respectively. While those in the ITS genes were identified with the nucleotide regions of 88th to 463rd and 51st to 274th. The two isolates showed IC50 value with (6.24±5.21) and (9.84±0.36) µg/mL) concentrations respectively at 28h. Reverse transcription – PCR indicated that these cells showed high level of expression for p53 mRNA. Conclusions The morphology and molecular analysis identified HF.1 and HF.2 to be Fusarium solani and Emericella nidulans; new isolates of anticancer producing fungi from Wady El-Natroon city in Egypt. Treatment with the two isolates caused P53 expression in Caco-2 cell line. These two isolates can be used as an anticancer agents.

875–884

Authors: AM Massoud, HA Shalaby, RM El Khateeb, MS Mahmoud, MA Kutkat

Objective To evaluate the effects of Mirazid® and myrrh volatile oil on adult Fasciola gigantica (F. gigantica ) under laboratory conditions. Methods The effects of oleoresin extract of myrrh (Mirazid®) and myrrh volatile oil on the surface morphology of adult F. gigantica following treatment in vitro had been determined by scanning electron microscopy. The results were compared with those observed in the fluke tegument following incubation in triclabendazole sulphoxide (TCBZ-SO), active form, (Fasinex®, Ciba-Geigy). Results Observations of the efficacy of Mirazid® oleoresin extract and myrrh volatile oil indicated that both products showed dose-dependent anthelmintic efficacy. The anterior half of the fluke was consistently more severely affected than the posterior half. The surface changes induced by Mirazid® oleoresin extract were less severe than those observed after exposure to either myrrh volatile oil or TCBZ-SO. Flukes showed swelling after these treatments, but its level and blebbing were much greater with myrrh volatile oil; in which patches of tegumental sloughing were observed in the apical cone and the posterior mid-body region of flukes. This was not observed after treatment with Mirazid® oleoresin extract. Conclusions The comparatively more disruption, observed in myrrh volatile oil exposed specimens, compared to that exposed to Mirazid® oleoresin extract might suggest that the anthelmintic activity of Mirazid® oleo resin extract was attributed to its content of volatile oil. So, increasing the concentration of myrrh volatile oil in Mirazid® might possibly help to developing its anthelmintic activity.

889–894

Authors: Mumuni Audu Momoh1, Charles Okechukwu Esimone

Objective To formulate gentamicin liposphere by solvent-melting method using lipids and polyethylene glycol 4 000 (PEG-4 000) for oral administration. Methods Gentamicin lipospheres were prepared by melt-emulsification using 30% w/w Phospholipon® 90H in Beeswax as the lipid matrix containing PEG-4 000. These lipospheres were characterized by evaluating on encapsulation efficiency, loading capacity, change in pH and the release profile. Antimicrobial activities were evaluated against Escherichia coli, Pseudomonas aeruginosa, Salmonella paratyphii and Staphylococcus aureus using the agar diffusion method. Results Photomicrographs revealed spherical particles within a micrometer range with minimal growth after 1 month. The release of gentamicin in vitro varied widely with the PEG-4 000 contents. Moreover, significant (P>0.05) amount of gentamicin was released in vivo from the formulation. The encapsulation and loading capacity were all high, indicating the ability of the lipids to take up the drug. The antimicrobial activities were very high especially against Pseudomonas compare to other test organisms. This strongly suggested that the formulation retain its bioactive characteristics. Conclusions This study strongly suggest that the issue of gentamicin stability and poor absorption in oral formulation could be adequately addressed by tactical engineering of lipid drug delivery systems such as lipospheres.

901–904

Authors: Watcharawit Rassami, Mayura Soonwera

Objective To determine the prevalence of infestation with head lice in primary schoolchildren in the eastern area of Bangkok, Thailand. Methods The present study was to determine the head lice infestation (Pediculosis) levels in primary schoolchildren, during May, 2011 to July, 2011, A total of 3 747 schoolchildren aged 5-12 years old from 12 selected primary school of Ladkrabang district, the eastern area of Bangkok were examined for head lice. Pediculosis was defined as the presence of at least on living adult, nymph and viable egg. Results The overall head lice infestation rate was 23.32% and infestation rate was higher in girls (47.12%) than in boys (0%). The infestation rate among schoolchildren varied from 12.62% to 29.76%. The infestation rate among girls varied from 26.07% (12 years old group) to 55.89% (8 years old group). Conclusions Pediculosis is a common public health problem affecting primary schoolchildren in eastern area of Bangkok and those levels are epidemic importance.

910–915

Authors: Viswanadha Vijaya Padma, Gurusamy Lalitha, Nicholson Puthanveedu Shirony, Rathinasamy Baskaran

Objective To assess the effect of quercetin (flavonoid) against lindane induced alterations in lipid profile of wistar rats. Methods Rats were administered orally with lindane (100 mg/kg body weight) and quercetin (10 mg/kg body weight) for 30 days. After the end of treatment period lipid profile was estimated in serum and tissue. Results Elevated levels of serum cholesterol, triglycerides, low density lipoprotein (LDL), very Low Density Lipoprotein (VLDL) and tissue triglycerides, cholesterol with concomitant decrease in serum HDL and tissue phospholipids were decreased in lindane treated rats were found to be significantly decreased in the quercetin and lindane co-treated rats. Conclusions Our study suggests that quercetin has hypolipidemic effect and offers protection against lindane induced toxicity in liver by restoring the altered levels of lipids. The quercetin cotreatment along with lindane for 30 days reversed these biochemical alterations in lipids induced by lindane.

923–924

Authors: Sadia Khan, Sujatha Sistla, Rahul Dhodapkar, Subhash Chandra Parija

Delftia acidovorans (earlier known as Comamonas acidovorans) is an aerobic, non-fermentative, Gram negative rod, classified in the Pseudomonas rRNA homology Group III. Reports of isolation of the organism from serious infections like central venous catheter associated bacteremia, corneal ulcers, otitis media exist. The microbiologists can identify this organism based on an orange indole reaction. This reaction demonstrates the organism's ability to produce anthranilic acid from tryptophan on addition of Kovac's reagent; which gives the media its characteristic “pumpkin orange” colour. Here we report the isolation of this organism from the Endotracheal tube aspirate of a 4 year old child. With the increasing use of invasive devices, it has become important to recognize these non fermentative gram negative bacilli as emerging source of infection even in immunocompetent individuals.

759–764

Authors: G Sarumathi, M Arumugam, S Kumaresan, T Balasubramanian

Objective To study the biological activities of the tissue extract of Cantharus tranquebaricus (C. tranquebaricus). Methods Crude extract of gastropod was tested for inhibition of bacterial growth. Antibacterial assay was carried out by disc diffusion method and the activity was measured accordingly based on the inhibition zone around the disc impregnated with gastropod extract. Molecular weight of the extract was determined by using SDS-PAGE. Plasma coagulation, Fibrin plate assay and substrate SDS-PAGE were used to determine the effect of sample on plasma coagulation, fibrin (ogen) olytic and proteolytic activity. Results The maximum inhibition zone (10 mm) was observed against Vibrio cholera (V. cholera) and minimum inhibition zone (2 mm) was noticed against Proteus mirablis (P. mirablis). The molecular weight was determined as 47-106 kDa. The tissue extract shows proteolytic activity above 48 kDa. SDS-PAGE analysis of fibrinogen after incubation with the tissue extract showed fibrinogenolytic activity. In plasma coagulation assay C. tranquebaricus tissue extract showed procoagulant property and it coagulated chicken plasma within 150 s, while control took 5 min to clot. The 9 HU hemolytic units were found against chicken blood and also exhibit high level of brine shrimp lethality. Conclusions This study suggests that C. tranquebaricus could be used as potential source for isolating bioactive compounds, since it is explored first time and found with promising results.

770–773

Authors: Sudha S, Masilamani Selvam M

Objective To investigate the cytotoxic activity of actinomycete isolated from marine sediment. Methods In the present study the DNA was isolated and the ITS region of 16s rRNA was amplified by polymerase chain reaction, using two universal bacterial primers, 1492R (5′-GGTTACCTTGTTAC GACTT-3′) and Eubac27F (5′-AGAGTTTGATCCTGGCTC AG-3′). The amplified products were purified using TIANgel mini purification kit, ligated to MD18-T simple vector (TaKaRa), and transformed into competent cells of Escherichia coli DH5α. 16S rRNA gene fragment was sequenced using forward primer M13F (-47) and reverse primer M13R (-48). Blast search sequence similarity was found against the existing non-redundant nucleotide sequence database thus, identified as Streptomyces sp SU, Streptomyces rubralavandulae strain SU1, Streptomyces cacaoi strain SU2, Streptomyces cavourensis strain SU3, Streptomyces avidinii strain SU4, Streptomyces globisporus strain SU5, Streptomyces variabilis strain SU6, Streptomyces coelicolor strain SU 7. Among the eight identified isolates, one actinomycete Streptomyces avidinii strain SU4 was selected for further study. Results Crude extract of the actinomycete isolate exhibited IC50 in 64.5 µg against Hep-2 cell line, 250 µg in VERO cell line. This value is very close to the criteria of cytotoxicity activity for the crude extracts, as established by the American National Cancer Institute (NCI) is in IC50 < 30 µg/mL. The GC MS analysis showed that the active principle might be 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (12.17%), isooctyl phthalate (15.29%) with the retention time 15.642 and 21.612, respectively. Conclusions This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs. These results help us to conclude that the potential of using metabolic engineering and post genomic approaches to isolate more bioactive compounds and make their possible commercial application is not far off.

782–786

Authors: Sasmal S, Majumdar S, Gupta M, Mukherjee A, Mukherjee PK

Objective To conduct a systemic evaluation of the medicinal value of seeds which include macroscopic and microscopic characterization, physiochemical evaluation, preliminary phytochemical screening and experimental antipyretic activity. Methods Saraca asoca seed was studied for pharmacognostical, phytochemical and other recommended methods for standardizations. Also, the acetone extract of the seeds was evaluated for acute toxicity study and antipyretic activity using Brewer's yeast induced pyrexia in Wistar rats at oral doses of 300 mg/kg and 500 mg/kg. Results After phytochemical screening, the acetone extract showed the presence of saponin, tannins and flavonoids which inhibit pyrexia. The therapeutic efficacy achieved at both the dose levels of the research drug and standard drug aspirin (100 mg/kg) showed significant (P<0.01) antipyretic activity when compared to the control group. The highly significant antipyretic effect exhibited at the dose of 500 mg/kg was also found to be sustainable in nature. Conclusions The antipyretic effect of the acetone extract showed significant results in rats at the dose of 500 mg/kg after following the standard pharmacognostical and phytochemical methods.

793–797

Authors: Ananthan G, Karthikeyan MM, Selva Prabhu A, Raghunathan C

Objective To investigate the seasonal fluctuations of the proximate composition of the ascidians muscle. Methods The moisture content was estimated by drying 1 g of fresh tissue at a constant temperature at 105 °C for 24 h.The loss of weight was taken as moisture content. The total protein was estimated using the Biuret method. The total carbohydrate in dried sample was estimated spectrophotometrically following the phenol- sulphuric acid method. The lipid in the dried sample tissue was gravimetrically estimated following the chloroform-methanol mixture method. Ash content was determined gravimetrically by incinerating 1 g dried sample in muffle furnace at about 550 °C for 6 h and results are expressed in percentage. Results It was found very difficult to compare the monthly variations, as all the ten species, exhibited wide fluctuations in their proximate compositions. For the sake of convenience, average seasonal values were calculated by summing the monthly values. Conclusions The proximate composition of the 10 commonly available ascidians showed high nutritive value and hence these groups especially solitary ascidians can be recommended for human consumption in terms of pickles, soup, curry and others after ensuring the safety of consumers.

803–810

Authors: Valan Arasu M, Asha KRT, Duraipandiyan V, Ignacimuthu S, Agastian P

Objective To isolate and indentify the promising antimicrobial metabolite producing Streptomyces strains from marine sediment samples from Andrapradesh coast of India. Methods Antagonistic actinomycetes were isolated by starch casein agar medium and modified nutrient agar medium with 1% glucose used as a base for primary screening. Significant antimicrobial metabolite producing strains were selected and identified by using biochemical and 16S rDNA level. Minimum inhibitory concentrations of the organic extracts were done by using broth micro dilution method. Results Among the 210 actinomycetes, 64.3% exhibited activity against Gram positive bacteria, 48.5 % showed activity towards Gram negative bacteria, 38.8% exhibited both Gram positive and negative bacteria and 80.85 % isolates revealed significant antifungal activity. However, five isolates AP-5, AP-18, AP-41 and AP-70 showed significant antimicrobial activity. The analysis of cell wall hydrolysates showed the presence of LL-diaminopimelic acid and glycine in all the isolates. Sequencing analysis indicated that the isolates shared 98.5%-99.8% sequence identity to the 16S rDNA gene sequences of the Streptomyces taxons. The antimicrobial substances were extracted using hexane and ethyl acetate from spent medium in which strains were cultivated at 30°Cfor five days. The antimicrobial activity was assessed using broth micro dilution technique. Each of the culture extracts from these five strains showed a typical polyene-like property. The lowest minimum inhibitory concentrations of ethyl acetate extracts against Escherichia coli and Curvularia lunata were 67.5 and 125.0 µg/mL, respectively. Conclusions It can be concluded that hexane and ethyl acetate soluble extracellular products of novel isolates are effective against pathogenic bacteria and fungi.

818–821

Authors: B. Ganga Rao, P. Umamaheswara Rao, E. Sambasiva Rao, T. Mallikarjuna Rao, and V. S. Praneeth. D

Objective To assess the in-vitro antibacterial activity and anti-inflammatory activity of orally administered different extracts (Hydro-alcoholic, methanolic, ethyl acetate and hexane) of Rauvolfia tetraphylla (R. tetraphylla) root bark in Carrageenan induced acute inflammation in rats. Methods In-vitro antibacterial activity was evaluated for extracts against four Gram positive and four Gram negative bacteria by using cylinder plate assay. Hydro-alcoholic extract (70% v/v ethanol) at 200, 400 and 800 mg/kg doses and methanolic, ethyl acetate and hexane extracts at doses 100, 200 and 400 mg/kg were tested for anti-inflammatory activity in Carrageenan induced rat paw oedema model and paw thickness was measured every one hour up to 6 hrs. Results All extracts of R. tetraphylla root bark showed good zone of inhibition against tested bacterial strains. In Carrageenan induced inflammation model, hydro-alcoholic and methanolic extract of R. tetraphylla root bark at three different doses produced significant (P<0.001) reduction when compared to vehicle treated control group and hexane, ethyl acetate extracts. Conclusions In the present study extracts of R. tetraphylla root bark shows good in-vitro antibacterial activity and in-vivo anti-inflammatory activity in rats.

826–829

Authors: Soundararajan Vijayarathna, Sreenivasan Sasidharan

Objective To investigate the cytotoxic effect of Elaeis guineensis methanol extract on MCF-7 and Vero cell. Methods In vitro cytotoxicity was evaluated in by MTT assay. Cell morphological changes were observed by using light microscope. Results The MTT assay indicated that methanol extract of the plant exhibited significant cytotoxic effects on MCF-7. Morphological alteration of the cell lines after exposure with Elaeis guineensis extract were observed under phase contrast microscope in the dose dependent manner. Conclusions The results suggest the probable use of the Elaeis guineensis methanol extract in preparing recipes for cancer-related ailments. Further studies on isolation of metabolites and their in vivo cytotoxicity are under investigation.

835–838

Authors: DK Patel, K Patel, V Tahilyani

Barbaloin is C-glucoside of aloe emodin anthrone which is found in the plant name as Aloe vera is a perennial succulent (Liliaceal), also called the healing plant. Barbaloin have variety of pharmacological activity such as strong inhibitory effect on histamine release, anti-inflammatory, cathartic, antiviral, antimicrobial, anticancer, antioxidant activity and alternative for pharmaceutical or cosmetic applications. The peak amount of barbaloin was reached about 3 h after oral administration. Concentration of barbaloin in Aloe vera leaves was shown to depend on the leaf part, age, and position of the leaf. Young leaves contain more barbaloin compared to old one. Various researches have been done on barbaloin but still the relationship between the barbaloin and its overall effect has not been clarified. A more specific perceptive of the pharmacological activities of barbaloin is required to develop for pharmaceutical purpose. Many attempts have been made regarding its isolation, biological activity to examine their effects, and clarify their functional mechanism. This review gives a brief idea about its uses, ethnomedicinally and commercially important analytical techniques and their pharmacological activities.