Asian Pacific Journal of Tropical Biomedicine

85–87

Authors: Juraj Majtán, Jaroslav Černy, Alena Ofúkaná, Peter Takáč, Milan Kozánek

Objective To investigate a case of mass mortality of Garra rufa (G. rufa) from a fish hatchery farm in Slovakia. Methods Causative bacterial agent was swabbing out of affected fish skin area and subsequently identified using commercial test system. Antibiotic susceptibility was determined by the disk diffusion method. Results Infected G. rufa was characterized by abnormal swimming behaviour, bleeding of skin lesions and local haemorrhages. Despite of using recommended aquatic antibiotic treatment no improvement was achieved and Aeromonas sobria (A. sobria) was identified as a causative agent of fish mortality. Due to massive fish mortality, antibiotic susceptibility of pure isolated culture of A. sobria was evaluated employing eight antibiotics against human infections. A. sobria was resistant only against one antibiotic, namely ampicilin. Conclusions These results indicate that A. sobria can act as a primary pathogen of G. rufa and may be a potential risk factor for immunodeficient or immunoincompetent patients during the ichthyotherapy.

92–96

Authors: S Ravikumar, M Fredimoses, M Gnanadesigan

Objective To investigate the anticancer property of marine sediment actinomycetes against two different breast cancer cell lines. Methods In vitro anticancer activity was carried out against breast (MCF-7 and MDA-MB-231) cancer cell lines. Partial sequences of the 16s rRNA gene, phylogenetic tree construction, multiple sequence analysis and secondary structure analysis were also carried out with the actinomycetes isolates. Results Of the selected five actinomycete isolates, ACT01 and ACT02 showed the IC50 value with (10.13±0.92) and (22.34±5.82) µg/mL concentrations, respectively for MCF-7 cell line at 48 h, but ACT01 showed the minimum (18.54±2.49 µg/mL) level of IC50 value with MDA-MB-231 cell line. Further, the 16s rRNA partial sequences of ACT01, ACT02, ACT03, ACT04 and ACT05 isolates were also deposited in NCBI data bank with the accession numbers of GQ478246, GQ478247, GQ478248, GQ478249 and GQ478250, respectively. The phylogenetic tree analysis showed that, the isolates of ACT02 and ACT03 were represented in group I and III, respectively, but ACT01 and ACT02 were represented in group II. The multiple sequence alignment of the actinomycete isolates showed that, the maximum identical conserved regions were identified with the nucleotide regions of 125 to 221st base pairs, 65 to 119th base pairs and 55, 48 and 31st base pairs. Secondary structure prediction of the 16s rRNA showed that, the maximum free energy was consumed with ACT03 isolate (-45.4 kkal/mol) and the minimum free energy was consumed with ACT04 isolate (-57.6 kkal/mol). Conclusions The actinomycete isolates of ACT01 and ACT02 (GQ478246 and GQ478247) which are isolated from sediment sample can be further used as anticancer agents against breast cancer cell lines.

102–103

Authors: Shiwen Wang, Jing He, Lijuan Zhang

Objective To evaluate the Anaplasma phagocytophilum (A. phagocytophilum), Ehrlichia canis (E. canis), Dirofilaria immitis (D. immitis) (canine heartworm), Borrelia burgdorferi (B. burgdorferi) infections in countryside dogs from Yunnan, Hainan and Anhui provinces. Methods Serum samples were collected from 26 dogs in Yunnan, Hainan and Anhui provinces. The samples were tested using a commercial ELISA rapid diagnostic assay kit (SNAP® 4Dx®; IDEXX Laboratories, Inc. U.S.A.). Meanwhile, indirect immunofluorescence assay (IFA) recommended by WHO was conducted to detect IgG to A. phagocytophilum. Two methods were analyzed and compared. Results The number of serologically positive dogs for IgG to A. phagocytophilum was only 2 which was from Hainan province and none of the 26 dogs responded positive for E. canis, D. immitis (canine heartworm), and B. burgdorferi by ELISA rapid diagnostic method. The number of serologically positive dogs for IgG to A. phagocytophilum was 13 (50%) by IFA method. Data of the two methods were analyzed by statistical software and the difference was statistically significant (P=0.002). Conclusions It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method. However, we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including A. phagocytophilum, E. canis, D. immitis (canine heartworm), and B. burgdorferi which have public health significance.

109–117

Authors: S Bhagavathy, P Sumathi

Objective To identify the available phytochemicals and carotenoids in the selected green algae and evaluate the potential genotoxic/antigenotoxic effect using lymphocytes. Methods Organic solvent extracts of Chlorococcum humicola (C. humicola) were used for the phytochemical analysis. The available carotenoids were assessed by HPLC, and LC-MS analysis. The genotoxicity was induced by the benzo(a)pyrene in the lymphocyte culture, the genotoxic and antigenotoxic effects of algal carotenoids with and without genotoxic inducer were evaluated by chromosomal aberration (CA), sister chromatid exchange (SCE) and micronucleus assay (MN). Results The results of the analysis showed that the algae were rich in carotenoids and fatty acids. In the total carotenoids lutein, β-carotene and α-carotene were found to be present in higher concentration. The frequency of CA and SCE increased by benzo(a)pyrene were significantly decreased by the carotenoids (P<0.05 for CA, P<0.001 for SCE). The MN frequencies of the cells were significantly decreased by the treatment with carotenoids when compared with the positive controls (P<0.05). Conclusions The findings of the present study demonstrate that, the green algae C. humicola is a rich source of bioactive compounds especially carotenoids which effectively fight against environmental genotoxic agents, the carotenoids itself is not a genotoxic substance and should be further considered for its beneficial effects.

125–127

Authors: Shakhawoat Hossain, Golam Kader, Farjana Nikkon, Tanzima Yeasmin

Objective To investigate the cytotoxicity of the crude ethanol extract of the rhizome of Zingiber zerumbet (Z. zerumbet) (L) Smith. and Curcuma zedoaria (C. zedoaria) Rosc. against Artemia salina Leach. Methods Fresh rhizomes of Z. zerumbet (L) Smith. and C. zedoaria Rosc. were extracted separately in cold with ethanol (2.5 L) and after concentration a brownish syrupy suspension of ethanol extracts of Z. zerumbet (L) Smith. and C. zedoaria Rosc. was obtained. The cytotoxic effect of the crude ethanol extracts of both plants was determined by brine shrimp lethality bioassay. Results Crude ethanol extracts of the rhizome of Z. zerumbet (L) Smith. showed the highest cytotoxicity (LC50 was 1.24 µg/mL) against brine shrimp nauplii as compared with C. zedoaria Rosc. (LC50 was 33.593 µg/mL) after 24 h of exposure. Conclusions It can be concluded that the rhizome of Z. zerumbet (L) Smith. and C. zedoaria Rosc. can be used as a source of cytotoxic agent.

134–139

Authors: Patel DK, Kumar R, Kumar M, Sairam K, Hemalatha S

To evaluate the aldose reductase inhibitory (ARI) activity of different fractions of Hybanthus enneaspermus for potential use in diabetic cataract. Methods Total phenol and flavonoid content of different fractions was determined. ARI activity of different fractions in rat lens was investigated in vitro. Results The results showed significant level of phenolic and flavonoid content in ethyl acetate fraction [total phenol (212.15±0.79 mg/g), total flavonoid (39.11±2.27 mg/g)] and aqueous fraction [total phenol (140.62±0.57 mg/g), total flavonoid (26.07±1.49 mg/g)] as compared with the chloroform fraction [total phenol (68.56±0.51 mg/g), total flavonoid (13.41±0.82 mg/g)] and petrolium ether fraction [total phenol (36.68±0.43 mg/g), total flavonoid (11.55±1.06 mg/g)]. There was a significant difference in the ARI activity of each fraction, and it was found to be the highest in ethyl acetate fraction [IC50 (49.26±1.76 µg/mL)] followed by aqueous extract [IC50 (70.83±2.82 µg/mL)] and it was least in the petroleum ether fraction [IC50 (118.89±0.71 µg/mL)]. Chloroform fraction showed moderate activity [IC50 (98.52±1.80 µg/mL)]. Conclusions Different fractions showed significanct amount of ARI activity, where in ethyl acetate fraction it was found to be maximum which may be due to its high phenolic and flavonoid content. The extract after further evaluation may be used in the treatment of diabetic cataract.

144–148

Authors: Jude E Okokon, Anwanga E Udoh, Samuel G Frank, Louis U Amazu

Objective To evaluate the anti-inflammatory and analgesic activities of leaf extract of Melanthera scandens (M. scandens). Methods The crude leaf extract (39–111 mg/kg) of M. scandens was investigated for anti-inflammatory and analgesic activities using various experimental models. The anti-inflammatory activity was investigated using carragenin, egg-albumin induced oedema models, while acetic acid, formalin-induced paw licking and thermal-induced pain models were used to evaluate the antinociceptive property. Results The extract caused a significant (P<0.05 – 0.001) dose-dependent reduction of inflammation and pains induced by different agents used. Conclusions The leaf extract possesses anti-inflammatory and analgesic effects which may be mediated through the phytochemical constituents of the plant.

152–155

Authors: Radhika Warikoo, Ankita Ray, Jasdeep Kaur Sandhu, Roopa Samal, Naim Wahab, Sarita Kumar

Objective To assess the larvicidal and irritant activities of the hexane extracts of leaves of Citrus sinensis (C. sinensis) against the early fourth instars and female adults of Aedes aegypti (Ae. aegypti). Methods The larvicidal potential of the prepared leaf extract was evaluated against early fourth instar larvae of Ae. aegypti using WHO protocol. The mortality counts were made after 24 h and LC50 and LC90 values were calculated. The efficacy of extract as mosquito irritant was assessed by contact irritancy assays. Extract-impregnated paper was placed on a glass plate over which a perspex funnel with a hole on the top was kept inverted. Single female adult, 3-day old unfed/blood-fed, was released inside the funnel. After 3 min of acclimatization time, the time taken for the first take-off and total number of flights undertaken during 15 min were scored. Results The citrus leaf extracts from hexane possessed moderate larvicidal efficiency against dengue vector. The bioassays resulted in an LC50 and LC90 value of 446.84 and 1 370.96 ppm, respectively after 24 h of exposure. However, the extracts were proved to be remarkable irritant against adults Ae. aegypti, more pronounced effects being observed on blood-fed females than unfed females. The extract-impregnated paper was thus proved to be 7–11 times more irritable as compared with the control paper. Conclusions The hexane extracts from C. sinensis leaves are proved to be reasonably larvicidal but remarkably irritant against dengue vector. Further studies are needed to identify the possible role of extract as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the extract could help in formulating strategies for mosquito control.

159–162

Authors: Orawan Khow, Sunutcha Suntrarachun

Bacteria have long been the favorite expression system for recombinant protein production. However, the flaw of the system is that insoluble and inactive proteins are co-produced due to codon bias, protein folding, phosphorylation, glycosylation, mRNA stability and promoter strength. Factors are cited and the methods to convert to soluble and active proteins are described, for example a tight control of Escherichia coli milieu, refolding from inclusion body and through fusion technology.

166–168

Authors: Suman S Karanth, Hariharan Regunath, Kiran Chawla, Mukhyaprana Prabhu

Burkholderia cepacia (B. cepacia) infection is rarely reported in an immunocompetent host. It is a well known occurence in patients with cystic fibrosis and chronic granulomatous disease where it increases both morbidity and mortality. It has also been included in the list of organisms causing nosocomial infections in an immunocompetent host, most of them transmitted from the immunocompromised patient in which this organism harbors. We report a rare case of isolation of B. cepacia from the bronchoalveolar lavage fluid of an immunocompetent agriculturist who presented with productive cough and fever associated with a pyopneumothorax. This is the first case of community acquired infection reported in an immunocompetent person in India.

464-469

Authors: Ramadoss Karthikeyan, PannemVenkatesh, Nesepogu Chandrasekhar

Objective: To evaluate the pharmacognostic parameters of the leaves of Abutilon indicum (Linn.) Sweet which will assist in standardization, quality assurance, purity and sample identification of the species. Methods: The pharmacognostic studies were carried out in terms of organoleptic, microscopic, powder microscopic, leaf constants and fluorescence analysis. Results: Macroscopic study showed that the leaf shape -cordate, Size -2-4 cm long, Colour - Green, Odour -Characteristic, Taste -Characteristic, Surface - Smooth, Apex -Acute to acuminate, Lamina-Simple, Cordate, Reticulate, Dentate, Margin-Crenate-Dentate. The microscopic features of leaves were observed as covering trichomes, glandular trichome, vascular bundles, crystals, stomata ,mucilage secretory cells adaxial epidermis, palisade mesophyll, spongy mesophyll and lateral vein. Further the study was evaluated leaf constants, powder microscopy and fluorescence study of leaf powder. Conclusion: Various pharmacognostic characters observed in this study help in the identification and standardization of Abutilon indicum (Linn.) Sweet.

477-483

Authors: Md. Sarfaraj Hussain, Sheeba Fareed, Mohammad Ali

Objective: A simple, accurate, and rapid high-performance thin-layer chromatographic (HPTLC) method for simultaneous quantification of the two biologically active flavonoidal compounds, gallic acid and quercetin, in Hygrophila auriculata (K. Schum) Heine (HA) has been established and validated. Methods: Chromatography was performed on aluminium foil-backed silica gel 60 F254 HPTLC plates with the binary mobile phase toluene: ethyl acetate: formic acid (5:4:1, v/v/v). Ultraviolet detection was performed densitometrically at the maximum absorbance wavelength, 270nm. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with ICH guidelines. Results: The system was found to give compact spots for gallic acid (GA) and quercetin (QE) (Rf value of 0.31 and 0.50, respectively). The limit of detection (23 and 41 ng band-1) limit of quantification (69 and 123 ng band-1), recovery (99.4-99.9 and 98.7-99.4%), and precision (i.e ≤1.98 and 1.97) were satisfactory for gallic acid and quercetin respectively. Linearity range for GA and QE were 100-1000 (r2= 0.9991) and 150-900 ng band-1 (r2= 0.9956) and the contents estimated as 0.28±0.01% and 0.41±0.01% w/w respectively. Conclusions: This simple, precise and accurate method gave good resolution from other constituents present in the extract. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing Hygrophila auriculata (K. Schum) Heine.

488-493

Authors: Pramod V. Pattar, M. Jayaraj

Objective: An efficient reproducible protocol has been developed for in vitro regeneration of plantlets from leaf and nodal explants of Aristolochia indica L. Methods: Wild grown plants Aristolochia indica L. were collected and grown in the departmental garden. Leaf and nodal segments (0.5-1.0 cm) from young healthy plants were first washed thoroughly under running tap water for 15 - 20 minutes and then treated with liquid detergent [5% (v/v) Tween-20] for 5-10 minutes. Later these explants were washed with double-distilled water for 5 minutes. Subsequently, explants were immersed in 70% (v/v) ethanol for 2 - 3 minutes and washed with sterile glass double distilled water for 2-3 times. Eventually, the explants were treated with an aqueous solution of 0.1% (w/v) HgCl2 for 1 - 2 minutes and rinsed for two-to-three times in sterile ddH 2O to remove all traces of HgCl2. The sterilized explants were inoculated aseptically onto solid basal Murashige and Skoog’s medium with different concentrations and combinations of BAP and NAA for in vitro regeneration of plants. Results: Both leaf and nodal explants cultured on MS medium supplemented with 0.8 mg/L BAP developed into mass of callus. These calli were subcultured for the induction of shoots and roots. Shoots were induced from both calli on MS medium supplemented with 0.8 mg/L BAP+0.5 mg/L NAA. Roots were induced from in vitro shoots on MS medium supplemented with 0.8 mg/L NAA for 4 weeks. Nodal explants were more regenerative with 95 % response compared to leaf explants with 85%. Finally, these in vitro regenerated plantlets were hardened, acclimatised and successfully transferred to the field. Conclusions: The present protocol for in vitro regeneration of Aristolochia indica L. can be used to make this plant available throughout the year for traditional healers, pharmaceutical usages, germplasm conservation, commercial cultivation, and also for the production of secondary metabolites.

501-505

Authors: Deenanath Jhade, Sachin Jain, Ankit Jain, Praveen Sharma

Objective: Pharmacognostic Screening and evaluate the in-vitro free radical scavenging activity of roots Acacia leucophloea. Methods: Pharmacognostic Standardization, Physico-chemical evaluation of the roots of Acacia leucophloea was carried out to determine its macro-and microscopical characters and also some of its quantitative standards. Microscopical studies were done by using trinocular microscope. Microscopically, root showed cork, cortex, stellar region and calcium oxalate crystals. Petroleum ether, ethanol, aqueous extracts of Acacia leucophloea were prepared, with successive extraction in soxhlet apparatus. Each extract was selected to study the free radical scavenging activity by superoxide scavenging assay method. Results: It was found that aqueous extract contained carbohydrates, glycosides amino acids flavonoids, tannins, alkaloids, steroids; ethanolic extract contained glycosides amino acids flavonoids, tannins, alkaloids, steroids. Ethanolic extract of Acacia leucophloea shows maximam inhibition in superoxide scavenging model. Aqueous extract also showed almost similar activity compared to ethanolic extract), while Petroleum ether extract showed poor inhibition of superoxide scavenging activity. Conclusion: The present study on pharmacognostic standardization, physico and phytochemical evaluation of Acacia leucophloea root might be useful to supplement information about its identification parameters assumed significantly in the way of acceptability of herbal drugs in present scenario lacking regulatory laws to control quality of herbal drugs.

511-513

Authors: P.K. Chauhan, V. Singh

Objective: To evaluate acute and subacute toxicity of the acetone extract of Centella asiatica (Brahmi). Methods: Toxicity of Centella asiatica was evaluated in Swiss mice after ingestion of the extract during one day (acute model) and during 15 days (subacute model). The Biochemical parameters evaluated included creatinine, calcium, inorganic phosphorous, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were assessed using commercial kits. Results: The results of the present investigation revealed that the LD50 of the extract is higher than 4000mg/kg and subacute treatment did not shows any change in corporal weight and hematological parameters. However, a change in liver weight but not in hepatic enzymes was observed. This suggested that the liver function is not altered by Centella asiatica. Some changes in the creatinine content were observed but could not be relative with the extract dose. Conclusions: The results suggest that the plant seems to be destitute of toxic effects in mice.

519-525

Authors: P Vijayabaskar, ST Somasundaram

Objective: To investigate the potent and specific anticoagulant activity of molluscan glycosaminoglycans (GAGs) isolated from whole clam tissue Donax cuneatus (D. cuneatus). Methods: Purification of few milligram quantities of GAGs from this tissue sample permitted a thorough examination of its anticoagulant activity characterization, which was partially purified by fractionation by anion exchange chromatography using DEAE cellulose column. The isolated crude and partially purified fractionated sample was showing metachromatic shift while using azure-A. The sample also exhibited prominent of biological and anti-fXa anticoagulant activity assays. Mobility was analyzed by two different buffer systems using agarose gel electrophoresis. Results: The fractionated molluscan GAG was also found to have similar peaks as that of standard heparin when assessed by the FT-IR spectrum. Finally molecular weight was determined by the gradient PAGE for crude and fractionated-II GAG, which were found to be 65 000 Da and 50 000 Da, respectively. Conclusions: The bivalve GAG was subjected to fractionation for further purification and its chemical components were analyzed. The fractionated clam heparin also showed substantial in vitro anticoagulant activity than that of commercial heparins.

532-535

Authors: Joghee Suresh, Jitin Ahuja, Nallupillai Paramakrishnan

Objective: To investigate the pharmacognostic characters of important medicinal plant, Artemisia parviflora (A. parviflora) Roxb. Methods: The pharmacognostic studies were carried out in terms of macroscopical, microscopical characters, physicochemical parameters of A. parviflora. Samples of different plant parts were fixed with formalin, acetic acid and ethyl alcohol (FAA), sectioned with rotary microtome and studied under microscope. Results: Some of the diagnostic features of the leaves and stems are the presence of amphistomatic epidermis, cyclocytic stomata, and collateral vascular bundles. Macroscopic study showed basal and lower stem leaves cuneate, oblong-obovate to flabellate, coarsely dentate to incised-dentate at the apex; middle and upper stem leaves mostly basally auriculate, palmatifid to deeply palmatisect or irregularly incised laciniate into linear to narrow lanceolate, acute segments. Further, ash and extractive values were calculated as per WHO guidelines. The preliminary phytochemical screening showed the presence of alkaloids, sterols/terpenoids, flavonoids, tannins, phenols and coumarins. Conclusion: Various pharmacognostical characters were observed in study which can help in identification and standardization of A. parviflora.

539-542

Authors: Narender Prasad D, B Ganga Rao, E Sambasiva Rao, T Mallikarjuna Rao, VS Praneeth D

Objective: To investigate the quantification of total phenolic, alkaloid content and in-vitro antioxidant activity of ethanol (70%), methanol, ethyl acetate and hexane extracts of Mesua ferrea (M. ferrea) leaves. Methods: The quantification of the total phenolic and alkaloid contents were estimated by taking gallic acid and atropine are as a standard; In-vitro antioxidant activity was evaluated for extracts by using different free radicals (superoxide, hydroxyl and DPPH). Results: M. ferrea leaves ethanol (70%) extract have more phenolic and alkaloidal content than other extracts. The selected plant extracts were produced concentration dependent percentage inhibition of different free radicals and produced maximum activity at a concentration of 1 280 µg and there after the percentage inhibition were raised gradually to its maximum level with higher concentrations. Conclusion: In the present study we found that the extracts of M. ferrea showed good antioxidant activity. Among the four extracts, the ethanol (70%) extract showed better activity than other extracts.

547-549

Authors: Keivan Saeb, Sara Gholamrezaee

Objective: To determine the best time of harvest for Melissa officinalis (M. officinalis) L. to gain highest amounts essential oil.Methods: M. officinalis leaves were harvested in three different stages (before flowering stage, flowering stage and after of flowering stage) and were dried. The essential oils were isolated by hydro- distillation and analyzed by GC/MS.Results: It showed that most essential oils of plants were in before flowering stage. In before flowering stage 37 compounds were identified in leaves oil of M.officinalis. The major components before flowering stage were decadienal (29.38%), geraniol (25.3%), caryophyllene oxide (8.75%), geranyl acetate (5.41%). In the flowering stage 36 compounds were identified as the major components of plant essential oils: decadienal (28.04%), geraniol (24.97%), caryophyllene oxide (7.55%), caryophyllene E (4.65%) and 16 components in the after flowering stage of plant were identified as the major components carvacrol (37.62%), methyl citronellate (32.34%), geranyl acetate (5.82%), caryophyllene (5.50%).Conclusions: The essential oils yields vary considerably from month-to-month and is also influenced by the micro-environment (sun or shade) in which the plant is growing. We found that the essential oil content of M. officinalis L. of leaves is significantly affected by harvesting stages.

555-558

Authors: R. Subramanian, P. Subbramaniyan, V. Raj

Objective: To determine the minerals, Iron (Fe), Sodium (Na), Potassium (K), Manganese (Mn) and trace elements, Lead (Pb), Copper (Cu), Cadmium (Cd) and Zinc (Zn) in two tropical medicinal plants (Syzygium caryophyllatum and Syzygium densiflorum). Methods: All the elements were determined by using atomic absorption spectrophotometer (AAS). Results: The average concentrations of Fe, Zn, Cu, Mn, Pb and Cd detected in leaves of Syzygium caryophyllatum and Syzygium densiflorum were 99.275±0.022, 17.302±0.010, 45.498±0.072, 47.344±0.021, 7.634±0.050, 0.253±0.003 and 93.829±0.015, 17.412±0.064, 64.866±0.058, 31.777±0.051, 6.768±0.010 and 0.343± 0.004, respectively. Conclusions: The mineral contents in the medicinal plants are at different levels. Therefore, these medicinal plants are rich in some essential elements and very least amount in trace elements, especially Fe and Mn which are essential for human health.

568-574

Authors: Krishnaveni Eahamban, Johnson Marimuthu, Antonisamy

Objective: The present study aimed to investigate the preliminary phytochemical analysis and UV-VIS, HPTC profiling and the antibacterial activity of Gracilaria corticata J. Ag extracts against the Gram positive and Gram negative bacteria. Methods: Preliminary phytochemical screening was carried out by Harborne method. The G. corticata extracts were tested against bacteria by the agar disc diffusion method. Results: The results of the presence study showed the presence of alkaloids, steroids, phenolic groups, saponins, tannin, flavonoids, terpenoids, glycosides and sugars. Proteins, xantoproteins, coumarins and catechin did not show any positive result for their presence in any of the six extracts of Gracilaria corticata tested. The result of the present study revealed the various behavior character of Gracilaria corticata crude drug. The UV-VIS spectrum profile of Gracilaria corticata methanolic, petroleum ether, benzene and aqueous extracts profiles were recorded. The HPLC profile of Gracilaria corticata petroleum ether benzene and aqueous extracts were tabulated. The maximum (9/12 bacterial pathogens) degree of antibacterial activity was observed in isopropanol soxhlet extracts followed by isopropanol cold extracts (7/12 bacterial pathogens). Conclusion: The results of the present study showed that G. corticata may be rich sources of phytoconstituents which can be isolated and further screened for different kinds of biological activities, depending on their reported therapeutic uses.

580-583

Authors: Akpera T. MANASSEH (FIMLS), Jombo T.A. Godwin (FMCPath), EMANGHE E Ubleni (FMCPath), Oguntayo O. Borisde (MSc)

Objective: This study was set up to review the phytochemistry of Ganoderma applanatum, its potentiality in nanotechological engineering for clinical use as well as impact of aqueous extracts of Ganoderma applanatum on laboratoery rats infected with Trypanosoma brucei brucei. Methods: Aqueous extracts of Ganoderma applanatum were obtained using hot sterile distilled water and whatmann filter paper. The presence of saponins, alkaloids, tannins, anthraquinones, flavonoids, cardiac glycosides and steroids were tested using standard procedures. Acqueous extracts were also inoculated into laboratory rats infected with Trypanosoma brucei brucei along with both positive and negative controls. Blood samples were collected daily, stained with Giemsa’s stain and examined for the presence of parasites.Results: Ganoderma applanatum aqueous extracts contained detectable levels of saponins, flavonoids, cardiac glycosides and steroids but undetectable levels of alkaloids, tannins and anthraquinone. All the infected rats died by day 12 from overwhelming trypanosomal infections.Conclusion: The biochemical constituents of Ganoderma species should be subjected to nanotechnological engineering in order to probably discover more of its wider therapeutic benefits, and to further disprove its suitability or otherwise in the treatment of African sleeping sickness.

589-592

Authors: SS Saboo, GG Tapadiya, SS Khadabadi

Objective: To study the morpho-anatomy of the leaf, stem and root of these plants to increase the knowledge and standardization parameters of Trichodesma sedgwickianum (TRS) and Trichodesma indium (TRI) of Borginaceae family.Methods: Transverse section of different organs of plants carried out by free hand and various types of histochemical tests were performed for the identification of microchemical by using different reagents.Results: Transverse section of TRS leaf shows presense of phloem tissue and anomocytic stomata with both glandular and covering trichomes. Stem contain epidermal brown tissue and collenchyma. Root shows presence of xylem and starch grains. TRI leaf contains radiating arc of xylem, phloem and pericyclic fiber. Stem shows presence of cortex with chlorenchymatous cells. Anatomically roots revealed the presense of xylem, phloem and oil globules. Unicerate covering trichomes and anisocytic stomata are present. Conclusions: The pharmacognostic constants and diagnostic microscopic features reported in this work could be useful for the compilation of a suitable monograph and its proper identification and standardisation of these two species.

601-604

Authors: Arokiyaraj S, Sripriya N, Bhagya R, Radhika B, Prameela L, Udayaprakash NK

Objective: To evaluate methanolic extracts of leaves of Artemisia nilagirica, Mimosa pudica and Clerodendrum siphonanthus for phytochemical analysis, antibacterial activity and free radical scavenging activity. Methods: Antibacterial activity was performed by disc diffusion method against two gram positive and four gram negative strains. Free radical scavenging potential was evaluated using total antioxidant activity (thiocyanate method) and diphenyl-picryl-hydrazyl (DPPH) assay. Results: Results of the present study showed that Clerodendrum siphonanthus exhibited significant antibacterial effect against Klebsiella pneumoniae (30 mm), Proteus mirabilis (16 mm), Salmonella typhi (16 mm), Staphylococcus aureus (12 mm), Escherichia coli (11.5 mm) and Bacillus subtilis (10 mm). Mimosa pudica and Artemisia nilagirica showed good antibacterial effects. Clerodendrum siphonanthus was found to be extremely effective in scavenging lipid peroxide (IC50 8 mg/mL) and DPPH radicals (IC50 7 mg/mL), whereas Artemisia nilagirica and Mimosa pudica showed moderate activity. Phytochemical analysis of these plants revealed presence of tannins, alkaloids, flavanoids, terpenoids and glycosides. Conclusions: This study showed that Artemisia nilagirica, Mimosa pudica and Clerodendrum siphonanthus may serve as a potential agent for new therapeutics.

612-617

Authors: Md Sarfaraj Hussain, Sheeba Fareed, Mohammad Ali

Objective: To analyse the two marker compounds lupeol (LP) and stigmasterol (ST) from methanolic extract of Hygrophila auriculata (H. auriculata). Methods: Separation was achieved on aluminium plates precoated with silica gel 60F254 with toluene-methanol-formic acid (7.0: 2.7: 0.3 v/v/v) as mobile phase. Results: Densitometric analysis was performed at 530 nm in the reflectance mode. Compact bands for LP and ST were obtained at RF 0.52 ± 0.02 and 0.28 ± 0.05. Linearity (r2=0.998 5 and 0.993 7), limit of detection (45 and 18 ng/band) limit of quantification (135 and 54 ng/band), recovery (98.2%-99.7% and 97.2%-99.6%), and precision (<2.18 and 1.91) were satisfactory for LP and ST respectively. Linearity range for LP and ST were 100-1 000 and 50-500 ng/band and the contents estimated as (0.19±0.1)% and (0.47±0.1)% w/w respectively. Conclusion: The method demonstrated efficient analysis testing of LP and ST in samples; therefore it can be used for routine analysis.

623-626

Authors: Deepak M Kasote, Anand A Zanwar, Santosh T Devkar, Mahabaleshwar V Hegde, Keshav K Deshmukh

Objective: To investigate the immunomodulatory activity of ether insoluble phenolic components of n-butanol fraction (EPC-BF) of flaxseed in rat. Methods: Immunomodulatory activity of EPC-BF was assessed in rat at oral doses of 150 and 300 mg/kg using neutrophil adhesion test, heamagglutination test and measuring delayed type hypersensitivity response. Results: EPCBF modulates immune response by significantly reducing cell-mediated immune response at oral dose 150 mg/kg but not at dose 300 mg/kg. However, EPC-BF did not affect on the neutrophil adhesion and humoral immunity at both the studied doses. Conclusion: EPC-PC modulates immune response by reducing cell-mediated immune response at lower dose but not at higher dose, which could be due to the presence of both immunostimulant and immunosuppressant phenols together in EPC-BF. These studies suggest that EPC-BF could be a promising source for new immune modifier.

632-636

Authors: Sanchita Das, Asis Bala, Manas Bhowmik, Lakshmi Kanta Ghosh

Objective: To investigate Schima wallichii (S. wallichii) Choisy (Ternstroemiaceae) which is a well known plant of Sikkim in the Himalayan region, India. Methods: Therefore three major flavonoid enriched fractions (FPet.Ether, FChloroform and FEthylacetate) were isolated by petroleum ether chloroform and ethyl acetate successively. The reactive nitrogen species scavenging activity of the flavonoid fractions was established using biochemical assay to measure scavenging of 2, 2 diphenyl picrylhydrazyl (DPPH), nitric oxide (NO) and peroxinitrite. Results: FEthylacetate showed maximum scavenging activity: their IC50 being (7.33 ± 3.32), (7.11 ± 2.21), and (6.67 ± 2.23) µg/mL in DPPH, NO, peroxinitrite radical respectively. Presence of (57.32 ± 2.31) and (163.4 ± 2.22) µg of flavonoids and phenolic compound in 1 mg of extract is assumed to be responsible for free radical scavenging activity. Conclusion: Taken together S. wallichii has potent free radical scavenging property indicating its importance in food supplement as a rich source of active flavonoid and phenolic compounds in ethyl acetate fraction which is responsible for its free radical scavenging as well as antioxidant activity.

641-645

Authors: Vinod Kumar Nathan, Johnson Marimuthu Antonisamy, Wesely Edward Gnanaraj, Kavitha Marappampalayam Subramanian

Objective: The present study was aimed to investigate the preliminary phytochemical analysis and HPTLC profiling and the antibacterial activity of P. pterocarpum methanolic flower extracts against the bacteria isolated from human infections. Methods: The preliminary phytochemical screening was performed according to the Harborne method. HPTLC studies were carried out using Harborne and Wagner et al method. The methanolic flower extracts of P. pterocarpum were tested against Salmonella typhi (MTCC 733), Staphylococcus aureus (MTCC 96), Proteus mirabilis (MTCC 742), Bacillus subtilis (MTCC 441) and Escherichia coli (MTCC 443). The antimicrobial activity was tested through well diffusion method. Results: The phytochemical studies on methanolic flower extract of Peltophorum pterocarpum (DC.) Baker ex Heyne. revealed the presence of glycosides, flavonoids, phenolics, saponins, catechin and alkaloids. The HPTLC separation was achieved using ethyl acetate-methanol-ethanol-water (8.1: 1.1: 0.4: 0.8) as the mobile phase. The methanolic extract of P. pterocarpum showed four different Rf values 0.16, 0.31, 0.77 and 0.82 which indicated various glycosides present in the flower extract. The methanolic extract of P. pterocarpum showed the maximum zone of inhibition against Proteus mirabilis followed by Salmonella typhi. Conclusion: Bio-assay revealed the presence of specific and selective antimicrobial compounds in the fractions. Broad range activity of plant extracts as per observations in this study was due to presence of multiple antimicrobial compounds or synergic effects of these compounds. Therefore, standardization of active fractions and study for in vivo efficacy may result in development of better antimicrobial drugs.

656-661

Authors: Monjoy Kumar Choudhury, S Venkatraman, Lokesh Upadhyay

Objective: To investigate the anti-hyperglycemic and peripheral glucose utilization activity of petroleum ether extract of leafs of Streblus asper (SAPE) as well as to find out the hypoglycemic property of Apiol and its presence in the SAPE. Methods: The GCMS analysis was conducted to identify the Apiol present in the SAPE. Six groups of animals, containing six animals in each, were selected for this study. The first group served as control and provided only vehicle solution. Group second served as vehicle control and treated with 0.5% CMC (carboxy methyl cellulose) orally. Group third to sixth, were treated with glibemclamide (600 ug/kg bw) and SAPE 100 mg/kg bw, 250 mg/kg bw and 500 mg/kg bw respectively. In the second experiment, three groups of animals containing six animals in each were selected and were made diabetic as described earlier and simultaneously treated with pure Apiol at the dosages of 50 mg/kg bw, 75 mg/bw and 100 mg/ kg bw respectively. Results: The GCMS analysis reviled the presence of Apiol molecules in the leaves of SAPE. Administration of SAPE and Apiol to the diabetic rats exhibited significant anti diabetic property (P<0.01). After 30 days of SAPE treatment, the fasting blood sugar levels of the diabetic rats were significantly reduced (P<0.001) along with restoration in their glycolytic and gluconeogenic enzyme activities, glycogen content and insulin level. Conclusions: The SAPE exhibited remarkable anti diabetic activity and excellent control over peripheral glucose utilization. Thus, it can be concluded that Apiol may be responsible for its anti-diabetic property.

667-669

Authors: Hema Lohani, Harish C. Andola. Nirpendra Chauhan, Ujjwal Bhandari

Objective: To evaluate the variation of Cinnamomum tamala leaf essential oil in respect with months and tree size class. Methods: By fallowing standard laboratory methods using capillary GC-FID and GC-MS. Results: The oil samples were analyzed for two months and two different tree size classes. cinnamaldehyde, was the principal component, was higher in the both months i.e. October and January, highest in 30 cm girth class tree in January month (59.23%) and lowest in 60 cm girth class tree in a month of October (41.90%). Smaller size class tree contained greater amount of cinnamaldehyde as compared to higher size class. Conclusions: Consequences of the present study need to selection of proper, tree size and month for harsh economic potentials of such valuable resources.

680-684

Authors: Kalpna Rakholiya, Sumitra Chanda

Objective: The aim of the present study was to investigate the morphological and microscopical characters of Mangifera indica L. leaf collected from Gujarat region and its phytochemical and physicochemical analysis. Methods: Microscopic characters and powder analysis was done under microscope. The physiochemical properties such as loss on drying, total ash value, acid insoluble ash value, water soluble ash value, pH, solubility and extractive values of M. indica were carried out. Results: The upper epidermis was irregular, wavy with thick epidermal layer; and prismatic, rosette and cluster types of calcium oxalate crystals were found. In phytochemical analysis, cardiac glycosides and tannins showed maximum amounts. Conclusions: The present study provides pharmacognostical, physicochemical and phytochemical details of the M. indica leaf which are useful in laying down standardization and pharmacopoeia parameters.

689-694

Authors: Saboo Shweta, Tapadiya Ganesh, Khadabadi Somshekhar

Objectives: To study the morpho-anatomy of the aerial parts of Rivea hypocrateriformi (Convolvulaceae) to increase the knowledge and standardization parameters of these plants and its family. Methods: Morpho-anatomical studies of aerial parts of have been carried out by free hand. The different types of histochemical tests were performed for the identification of micro-chemical by using staining reagents. Preliminary phytochemical and quantitative estimation has been determined along with HPTLC fingerprinting. CAMAG HPTLC system equipped with Linomat V applicator, TLC scanner 3, Reprostar 3 and WIN CATS-4 software was used for fingerprinting. Results: The morpho-anatomical structure of plant revealed the presence of thick wall collenchyma cell and parenchyma cells along with spharaphides and columnar palisade cells. Presense of numerous covering trichomes, parasitic stomata, oval shaped vascular bundles and calcium oxalate crystals were observed. In powder microscopy lignified xylem vessels, and covering trichomes were clearly seen. Physicochemical parameters, ash value, inorganic elements, moisture content and extractive value were determined to develop the stringent pharmacognostic standards. Qualitative standardization and HPTLC fingerprinting of phytochemical revealed the presense of various plant secondary metabolites. Conclusions: These findings will be useful in establishing pharmacognostic and phytochemical standards for identification, as well as assessment of purity and quality of this plant, which definitely gaining the relevance in plant drug research and establishment of plant monograph.

700-705

Authors: Smilin Bell Aseervatham G, Shamna R, Sangeetha, Sasikumar JM

Objective: To evaluate the in vivo activity of bark extract of Bixa orellana L. (B. orellana) against acetaminophen induced oxidative stress. Methods: In the present study, antioxidant activity of B. orellana was evaluated by using normal and acetaminophen induced oxidative stressed rats at the dose of 100 mg/kg and 200 mg/kg p.o. oraly daily for 20 days. The animal's body weight was checked before and after treatment. Different biochemical parameters such as serum glutamate pyruvate transaminases, serum glutamate oxalo transaminases, alkaline phosphatase, total bilirubin, cholesterol, protein, lactate dehydrogenase, superoxide dismutase, catalase, ascorbic acid, lipid peroxide was performed. Histopathological analysis of the control and the hepatotoxicity induced rats were performed. Results: It was observed that the B. orellana bark extract showed significant protective activity against acetaminophen induced damage at 200 mg/kg dose level, while the 100 mg/kg dose showed moderate activity. Conclusions: From the result obtained in the present study suggest that B. orellana bark extract elicit protective activity through antioxidant activity on acetaminophen induced hepatic damage in rats.

712-715

Authors: Nidhi Gupta, Aditya Ganeshpurkar, Nishikant Jatav, Divya Bansal, Nazneen Dubey

Objective: To investigate chick pancreatic lipase inhibitory activities of the Abroma augusta (A. augusta). Methods: A. augusta was first extracted with methanol and subjected to phytochemical screenings. Quantitative estimation of flavonoids, phenolics and alkaloids was done. Pancreatic lipase from chick pancreas was isolated and used as substrate for anti-lipase studies. Results: A. augusta extract effectively inhibited concentration dependent lipase activity, whereby extract at concentration 100 µg/mL inhibited 88.6% enzyme activity. Conclusions: From these results, it could be concluded that A. augusta can be used as a potential source anti-lipase agents.

723-726

Authors: Jimuty Roy, Mohammad Kuddus, Bilkis Begum, Choudhury Hasan

Objective: To investigate the crude methanolic extract of whole plant part of Sansevieria roxburghiana Schult. and Schult. f. (Agavaceae) and the possibility of analgesic, cytotoxic and antioxidant activities. Methods: The analgesic activity was assessed by acetic acid induced writhing test in mice. The cytotoxic activity was evaluated by brine shrimp lethality bioassay while antioxidant effect was measured by 1, 1-diphenyl-2-picrylhydrazyl-hydrate (DPPH) free radical scavenging assay. Results: The ethyl acetate soluble fraction of the crude extract was fo und to have significant (P<0.001) analgesic activity at the oral dose of 100 mg/kg body weight. In brine shrimp lethality bioassay, the aqueous soluble fraction exhibited maximum toxicity towards the shrimp with LC50 value of 0.735 µg/mL compared to 0.544 µg/mL exhibited by standard vincristine. The crude methanolic extract along with its all partitionates revealed mild to moderate free radical scavenging activity. Conclusions: These primary findings suggest that the extract might possess some chemical constituents that are responsible for analgesic, cytotoxic and antioxidant activities.

732-736

Authors: Thambiraj J, Paulsamy S

Objective: The in vitro antioxidant capability of methanolic leaf extract of the medicinal plant, Acacia caesia (L.) Willd (Mimosaceae) was evaluated by different assaying. Methods: The in vitro antioxidant activity was evaluated for total antioxidant, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, hydroxyl radical scavenging and metal chelating activities. Ascorbic acid and butylated hydroxy toluene (BHT) were kept as standards. Results: IC50 values observed fo r DPPH, hydroxyl radical scavenging and metal chelating assays were 109, 177 and 295µg/ ml respectively. Conclusions: The results clearly indicate that the methanolic leaf extract of the study species is effective in scavenging free radicals and has the potential to be powerful antioxidant.

741-749

Authors: Malairaj Sathuvan, Anadhan Vignesh, Ramar Thangam, Perumal Palani, Ramasamy Rengasamy, Kandasamy Murugesan

Objective: To evaluate the antioxidant and anticancer potential of different fractions of bark of Costus pictus using various in vitro antioxidant assay systems. Methods: In this study, assay like DPPH radical, superoxide anion radical scavenging activity, nitric oxide scavenging activity, hydrogen peroxide scavenging activity, metal chelating activity and reducing power were used. The concentrations of total phenolic and flavonoids were also calculated for the extracts. Result: The present study elucidated for the first time the antioxidant property of bark of C. pictus. This study suggested that, among the three fractions, the chloroform fraction possesses high antioxidant activity which might be helpful in preventing or slowing the progress of various oxidative stress related disorders. Moreover, all fractions possess potent anticancer properties against colon cancer cells of HT29 and lung carcinoma cells of A549. Conclusions: It can be concluded that the extract of the bark of C. pictus has potential natural antioxidant and this can be used in food industries. There are few reports on the antioxidant capacity of bark of C. pictus and the mechanism of different fractions of bark of C. pictus as antioxidative agents is still not fu lly understood. Hence further research is underway to analyse and isolate the active compounds responsible for the antioxidant and anticancer activity of different fractions of the bark of C. pictus.

753-756

Authors: Subash vijayakumar, Ramaiyan Dhanapal, I. Sarathchandran, A. Saravana Kumar, J.Vijaya Ratna

Objective: To study the effect of antioxidant activity of the ethanol extract of whole plant of Ammania baccifera, Linn (EEAB). Methods: To investigate in rats against carbon tetrachloride (CCl4) induced erythrocyte damage. Simultaneous intraperitoneal administration of the crude extracts (600 or 800 mg/kg body weight/day) with carbon tetrachloride (1ml/kg of body weight) to rats for alternate days of two weeks protected the loss of functional integrity and membrane lipid alteration in red blood cells induced by oxidative stress. Results: EEAB inhibited the accumulation of lipid peroxidation products in the plasma as well as maintained the activities of antioxidant enzymes such as superoxide dismutase (SOD) and catalase. The extracts further had the ability to decrease the membrane fluidity induced by carbon tetrachloride. Conclusion: It can therefore be suggested that the whole plant of Ammania baccifera, Linn possess an erythrocyte protective activity against drug induced oxidative stress. These findings also provide a rationale for further studies on isolation of active principles and its pharmacological evaluation.

765-771

Authors: Ayyakkannu Purushothaman, Elumalai Nandhakumar, Panchanatham Sachdanandam

Objective: To investigate the therapeutic efficacy of a Shemamruthaa (SM), (combination of Hibiscus rosasinensis (H. rosasinensis) flowers, fruits of Phyllanthus emblica (P. emblica) and pure honey in definite ratio), against lipid peroxidation (LPO) and antioxidant status in experimentally induced mammary carcinoma rats. Methods: Adult female Sprague-Dawley rats were used for the study and were divided into four groups. Group I control animals received standard pellet diet and water ad libitum. Group II rats were induced with 7,12-dimethyl benz[a]anthracene (DMBA) (25 mg in 1 mL olive oil) by gastric intubation, whereas another set of DMBA-induced rats were treated with SM (400 mg/kg body weight/d) in olive oil orally by gastric intubation for 14 d after 3 months of induction period (group III). Group IV rats served as SM-treated control animals. At the end of the experimental period, the rats were anaesthetised and sacrificed and used for biochemical measures and histology studies. Results: The LPO was increased and antioxidant levels were decreased in the serum, liver and mammary tissues of cancer-induced rats. The administration of SM drug significantly (P<0.05) decreased LPO and reversed the status of antioxidants to near normal level in cancer-bearing animals. Conclusions: The results obtained indicate the additive and synergistic action of constituents’ plants in the SM drug against oxidative damage and its protective role in DMBA induced mammary cancer.

781-787

Authors: Manish Kumar, Madhu Chandel, Neha Sharma, Subodh Kumar, Satwinderjeet Kaur

Objective: To investigate the antioxidant and genoprotective potential of the methanol extract (METH-KP) along with its hexane fraction (HEX-KP) from the leaves of Koelreuteria paniculata (K. paniculata) Laxm. Methods: The antioxidant potential was checked using metal chelation assay, ABTS, DPPH (2,2-diphenyl-2-picryl-hydrazyl), reducing power and superoxide anion radical scavenging assay, Folin-Ciocalteu reducing capacity (FCR assay), Total flavonoid content (TFC assay) and genoprotective activity against the DNA damage induced by Fenton's reagent using pUC18/calf thymus. Results: Results showed that radical scavenging activities of the both test extract/fraction revealed a concentration-dependent antiradical activity in all the assays. In Metal chelation assay, METH-KP and HEX-KP showed 39.04% and 32.51% of scavenging at highest tested concentrations. The METH-KP exhibited IC50 of 54.54 µg/mL in ABTS, 115 µg/mL in DPPH, 110 µg/mL in reducing power and 135 µg/mL in superoxide anion radical scavenging assay while that HEX-KP was found to be very poor in radical scavenging in all the above assays. The phytochemical analysis showed good amount of phenolic and flavonoid compounds in METHKP while the HEX-KP fraction lacks phenols. The METH-KP extract and HEX-KP fraction both showed DNA protective effect in Calf thymus/pUC18 DNA protection studies. Conclusions: The activity of METH-KP may be attributed to its polyphenolic constituents which needs further isolation of its active constituents which may lead to the development of novel drugs to combat cancer.

796-799

Authors: Supaluk Prachayasittikul, Apilak Worachartcheewan, Sakda Yainoy, Natthakaln Lomchoey, Paveena Kittiphatcharin, Somsak Ruchirawat, Virapong Prachayasittikul

Objective: To investigate antioxidant and antimicrobial activities of Saraca thaipingensis Cantley ex Prain; and isolation of its flower extracts. Methods: The plant species (flowers, leaves, and twigs) were extracted by hexane, dichloromethane, ethyl acetate and methanol; and tested for antioxidant activity (DPPH assay) and antimicrobial activity (agar dilution method) against twenty-seven strains of microorganisms; gram positive and gram negative bacteria, and diploid fungus. Bioactive constituents were isolated by column chromatography. Results: The plant extracts has been firstly reported to display strong antioxidant activity and antimicrobial activity selective against gram positive bacteria (Corynebacterium diphtheriae NCTC 10356 and Streptococcus pyogenes) with MIC of 256 µg/mL. Stigmasterol and a mixture of triterpenoids and phenolic compounds were isolated from the flower extracts. Conclusions: The study revealed that the S. thaipingensis is a new source of natural antioxidants and antimicrobials with potential fo r medicinal uses.

808-811

Authors: Sharma KK, Kotoky J, Kalita JC, Barthakur R

Objective: To investigate the in vitro antifungal activity of Ranunculus sceleratus (R. sceleratus) and Pongamia pinnata (P. pinnata), collected from North East India. Chloroform, Methanol and Water extracts of the plants were evaluated for anti-ringworm activity against five strains of dermatophytes viz., Trichophyton rubrum (T. rubrum), Trichophyton mentagrophytes (T. mentagrophytes), Trichophyton tonsurans (T. tonsurans), Microsporum gypseum (M. gypseum) and Microsporum fulvum (M. fulvum). Methods: Agar well diffusion method was used to determine the inhibition zone of the different extracts. The minimum inhibitory concentrations of the extracts were determined by broth macro dilution method. Results: When compared, the chloroform extracts of both the plants were found to be highly active against all the tested fungi fo llowed by the methanol extract. The water extract showed the least activity. The minimum inhibitory concentration (MIC) of the extracts was ranging from 1.25-10.00 mg/mL for chloroform and methanol extracts and greater that 10 mg/mL for the water extracts. Conclusion: The results obtained from the experiments suggest the anti-dermatophytic nature of the plants.

818-829

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

Objective: To evaluate antimicrobial potencies of 25 plants with reports on ethnomedicinal uses fo r infectious ailments by the aborigine Kandha tribe of Kalahandi district, Odisha state, India fo r urinary tract infections. Methods: Over a period of 6 months, multidrug resistant (MDR) strains of 6 uropathogenic bacteria Acinetobacter baumannii (A. baumannii), Citrobacter freundii (C. freundii), Klebsiella oxytoca (K. oxytoca), Proteus mirabilis (P. mirabilis), Proteus vulgaris (P. vulgaris) and Pseudomonas aeruginosa (P. aeruginosa) were isolated from clinical samples in a teaching hospital; their antibiograms were ascertained. Concentrated aqueous and ethanolic extracts of leaves and barks of plants were used for monitoring their antimicrobial potencies, by the agar-well diffusion method. Phytochemical analyses of plant parts were done. Results: All isolated bacterial strains were resistant to 15 antibiotics of 6 groups including β-lactams. From a surveillance of bacterial isolates, it was evident that the distribution of MDR strains of each was more in hospital acquired isolates than the community acquired ones. Both aqueous and ethanolic extracts of plants, Aegle marmelos (A. marmelos), Azadirachta indica (A. indica) and Withania somnifera (W. somnifera) were highly effective against MDR isolates of all these pathogens. Several plants were moderately effective during in vitro control of the pathogens. Plants, Anthocephalus cadamba (A. cadamba), Cleistanthus collinus (C. collinus) and Oroxylum indicum (O. indicum) were totally ineffective in the control of isolated MDR uropathogen. A. indica, T. arjuna and T. alata contained the full range of phytochemicals (alkaloids, glycosides, terpenoids, reducing sugars, saponins, tannins, flavonoids and steroids), which could be attributed to the significant anti-uropathogenic activities. Conclusion: Plants, A. indica, A. marmelos, Cassia fistula (C. fistula), T. arjuna, Salvadora persica (S. persica), W. somnifera and Vitex negundo (V. negundo), particularly could be useful for an use as complementary/ supplementary medicines for MDR uropathogens.

838-845

Authors: P Srinivas, S Ram Reddy

Objective: To investigate the antibacterial principle and activity of Aerva javanica, a medicinal plant. Methods: Crude extracts of different parts of Aerva javanica were made with hexane, chloroform and methanol. Phytochemical analysis of the crude extracts was done by following the standard methodology, and antibacterial activity was evaluated by inhibition zone and MIC values. Crude extracts were resolved through HPTLC and the antibacterial activity of the separated compounds was evaluated by bioautography. Results: The yields of crude extracts made from different plant parts varied both with plant part and solvent. Methanolic extracts of leaf and flower have shown a wide range of phytochemicals and more antibacterial activity. HPTLC separation of extracts coupled with bioautography studies revealed that apigenin followed by rutin and kaempferol has shown antibacterial activity against more number of bacteria. Conclusions: The present study supports the use of Aerva javanica in the traditional medicine, and it can be used against bacterial infections.

855-858

Authors: Saravana Kumar Arthanari, Jayachandran Vanitha, Mani Ganesh, Krishnasamy Venkateshwaran, De Clercq

Objective: To investigate the cytotoxicity and antiviral activity of methanolic extract of S. grandiflora flowers using different cell lines and viruses. Methods: The methanolic flower extracts were prepared and evaluated for their antiviral and cytotoxic activities using viruses like herpes simplex-1 and 2, vaccinia, vesicular stomatitis, cox sackie, respiratory syncytical, fe line corona, feline herpes, para influenza, reo-1, sindbis and punta toro viruses in different cell lines, like Hel, HeLa, Crandell Reus feline kidney and Vero cell cultures. Results: Among the viruses used the extract possessed strongest antiviral activity against herpes simplex 1 and 2, repiratory syncytical, para influenza, reo, sindbis, cox sackie and punta toro viruses that was (EC50 =20 µg/mL and 45 µg/mL) and moderate activity for remaining viruses (EC50= 100 µg/mL). The antiviral activities assessed by calculating the selectivity index may be due to the presence of fl avonoids in the extracts there by inhibit the virus cell fusion in the early and replication stages. The cytotoxicity effect was evaluated using MTT assay and the results revealed that the extracts exhibited cytotoxicity from the range of 20 to 100 µg/mL. Conclusions: Present results confirmed that the S. grandiflora used as a good antimicrobial agent in future.

863-869

Authors: Venkatachalam Uthayakumar, Venkatachalam Ramasubramanian, Dhanabalan Senthilkumar, Venkatesan Brindha Priyadarisini, Ramasamy Harikrishnan

Objective: To characterize the biochemical, antimicrobial and hemolytic activities of Mastacembalus armatus skin mucus. Methods: Antimicrobial and antifungal activities of mucus extractions against human and fish pathogens were tested along with ampicillin as control. Hemolytic activity of the extraction was evaluated against sheep and cow blood cells. Amino acid and fatty acid profiles were analyzed by HPLC and gas chromatography in the mucus of fish. SDSPAGE analysis of mucus and muscle tissue was done. Oneway-ANOVA was performed against all extraction and pathogens, amino acids and fatty acids. Result: All the mucus extracts exhibited higher inhibitory activity than antibiotic ampicillin against bacterial and fungal pathogens. The hemolytic activity was increased with higher mucus concentrations in both sheep and cow blood cells. The protein content soluble and insoluble fractions of mucus were 63.22 µg/g and 55.79 µg/g, respectively. Out of 17 amino acids leucine was higher (8.54 mole %) in soluble gel, and glutamic acid was higher (6.92 mole %) in the insoluble gel, Histidine was very low (i.e. 0.20 mole %) both in soluble and (0.30 mole %) insoluble gel. In SDS-PAGE analysis, 6 bands of mucus and 9 bands of muscle were observed. Conclusions: The soluble and insoluble proteins are responsible fo r antimicrobial and hemolytic activity, these results indicate that mucus gel was prospective applications in fish and human therapeutics.

876-880

Authors: Kalpna Rakholiya, Sumitra Chanda

Objective: To evaluate the in vitro interaction between methanolic extracts of Terminalia catappa (T. catappa) (Combretaceae) and Carica papaya (C. papaya) (caricaceae) leaves and certain known antimicrobial drugs like penicillin G (P), ampicillin (AMP), amoxyclav (AMC), cephalothin (CEP), polymyxin B (PB), rifampicin (RIF), amikacin (AK), nilidixic acid (NA), gentamicin (GEN), chloramphenicol (C), ofloxacin (OF) against five Gram positive and five Gram negative bacteria. Methods: Evaluation of synergy interaction between plant extracts and antimicrobial agents was carried out using disc diffusion method. Results: The results of this study showed that there is an increased activity in case of combination of methanolic plant extracts and test antimicrobial agents. The more potent result was that the synergism between methanolic extract of C. papaya and antibiotics showed highest and strong synergistic effect against tested bacterial strains; though methanolic extract of C. papaya alone was not showing any antibacterial activity. Conclusions: These results indicate that combination between plant extract and the antibiotics could be useful in fighting emerging drug-resistance microorganisms.

884-887

Authors: Shashi Kant Singh, Rupali M Tiwari, Saurabh K Sinha, Chhanda C Danta, Satyendra K Prasad

Objective: To isolate the mangiferin from Mangifera indica (M. indica) and assess the antimicrobial activity of different analogues synthesized from mangiferin. Methods: Mangiferin was isolated by column chromatography from the ethanolic extract of stem bark of M. indica. Mangiferin was further converted to 5-(N-phenylamino methyleno) mangiferin, 5-(N-pchlorophenylamino methyleno) mangiferin, 5-(N-2-methyl phenylamino methyleno) mangiferin, 5-(N-p-methoxy phenylamino methyleno) mangiferin, 5-(N, N-diphenylamino methyleno) mangiferin, 5-(N-α-napthylamino methyleno) mangiferin and 5-(N-4-methyl phenylamino methyleno) mangiferin. Mangiferin and its analogues were characterized by melting point and Rf value determination and through spectral technique like UV, IR, and NMR spectral analysis. The antimicrobial effect of mangiferin and its derivatives was studied according to the disc diffusion method. Results: The solutions of mangiferin and its derivatives in polyethylene glycol-400 showed an activity with regard to four bacterial species-Bacillus pumilus (B. pumilus), Bacillus cereus (B. cereus) and Salmonella virchow (S. virchow) and two fungal species-Thermoascus aurantiacus (T. aurantiacus) and Aspergillus flavus (A. flavus). Conclusion: The present study confirms the antimicrobial activity of different analogues of mangiferin which could be further processed for the development of a potential antimicrobial agent.

892-894

Authors: Alwar Vidyalakshmi, Kandaswamy Kruthika

Objective: To test efficacy of Parmelia perlata (P. perlata), which is used in traditional medicine fo r rapid wound healing against test bacteria that cause wound infections. Methods: Different solvents such as methanol, ethyl acetate and acetone were used for extraction of P. perlata. The sensitivity of the test bacteria to solvent extracts of P. perlata was tested by measuring the zone of inhibition on growth media and by determining the minimal inhibitory concentration and minimal bactericidal concentration. Results: Methanol, ethyl acetate and acetone extracts of P. perlata have shown inhibitory activity against Staphylococcus aureus (S.aureus). Conclusions: The results of the present study indicate that P. perlata has potential antibacterial compounds against S.aureus that causes multitude of skin infections among human beings. Development of drugs fr om natural compounds can help us to combat antibiotic-resistant bacteria.

901-904

Authors: M. Karunai Raj, V. Duraipandiyan, P. Agastian, S. Ignacimuthu

Objective: To investigate the antibacterial and antifungal properties of crude methanol extract of Peltophorum pterocarpum (P. pterocarpum) flower. Methods: Disc diffusion method was used fo r preliminary screening of extracts against microbes. The micro-dilution method was used fo r the determination of the minimal inhibition concentration (MIC). The isolated compound was identified by crystallography (XRD) technique. Results: Bergenin (C-glycosyl benzoic acid) was isolated from methanol fraction of P. pterocarpum flowers and confirmed using X-ray crystallography. It was tested against bacteria and fungi. Bergenin showed antifungal activity against Trichophyton mentagrophytes (T. mentagrophytes) (MIC 250 µg/mL), Epidermophyton floccosum (E. floccosum) (MIC 500 µg/mL), Trichophyton rubrum (T. rubrum) (MIC 500 µg/mL), Aspergillus niger (A. niger) (MIC 500 µg/mL) and Botrytis cinerea (B. cinerea) (MIC 250 µg/mL). No antibacterial activity was seen. Conclusion: The present study was helpful to discovery of new drug and complimentary of alternative medicine.

910-913

Authors: T.Thirumalai, Beverly C David, K.Sathiyaraj, B.Senthilkumar, E David

Objective: To conduct an ethnobotanical survey and to collect information from local people of Javadhu hills about the use of traditional medicinal plants diabetes treatment. Methods: Javadhu hills were surveyed through interviewing randomly selected 312 local participants using semi-structured questionnaire and regular field visits. Results: The investigations revealed that about 40 traditional plant species and their local names with parts used for the treatment were recorded by the local people of Javadhu hills. Conclusions: The study led to the abundant knowledge of wealth of traditional medicinal plants that are being used for the diabetes treatment by the local people of Javadhu hills.

919-923

Authors: Sunil Jawla, Y Kumar, MSY Khan

Objective: Bougainvillea spectabilis (B. spectabilis) is one of the main constituent of various herbal formulations available for diabetes. The aim of present study was to screen hypoglycemic potential of B. spectabilis stem bark extracts in albno rats (Wistar strain). Methods: The EtOH extracts (100, 250 and 500 mg/kg/day) of the B. spectabilis were administered to both normal and alloxan induced diabetic rats at defined time intervals. Blood glucose levels were measured at 0, 0.5, 1, 2, 4, 6 h, and on 0, 1, 3, 5, 7th day after oral administration of extracts. Of the doses test, highest anti-hyperglycemic effect was observed by the extract of stem bark at 250 mg/kg after a week treatment. Results: B. spectabilis stem bark extract exhibited significant hypoglycemic activity at different doses and intervals. Stem bark extract was found to be 22.2% more potent than standard oral hypoglycemic drug, glibenclamide 0.2 mg/kg. Conclusion: Treatment of alloxan induced diabetic rats up to a week with stem bark extract reversed the permanent hyperglycemia. Hence, B. spectabilis stem bark alcoholic extract exhibited potent hypoglycemic activity.

930-933

Authors: R Karthic, S Nagaraj, P Arulmurugan, S Seshadri, R Rengasamy, K Kathiravan

Objective: To study the antidiabetic effects of suspension cell extract of Gymnema sylvestre (G. sylvestre) along with field grown and wild plants. Methods: The effect of ethanolic extracts of the in vitro grown suspension cells of G. sylvestre along with field grown and wild plant leaves of G. sylvestre was tested on alloxan induced diabetic rats. Results: While oral administration of the extracts reduced the glucose content in blood and urine, sugar and lipids in serum significantly (P≤0.05), it also increased the body weight, total haemoglobin and plasma protein content. Conclusions: It can be concluded that G. sylvestre suspension cell extract show excellent antidiabetic potential against alloxan induced diabetic albino male rats therefore be considered as potent antidiabetic drug.

941-946

Authors: David Hansi Prisilla, Rangachari Balamurugan, Harshit R Shah

Objective: To investigate the antihperglycemic activity of methanol extract of Acorus calamus (AC) rhizome in streptozotocin (STZ) induced diabetic rats. Methods: Oral glucose tolerance test (OGTT) was performed in normal rats. Male albino rats were rendered diabetic by STZ (40 mg/kg, intraperitoneally). 200 mg/kg of AC extract was administered orally to diabetic rats for 21 days to determine the antihyperglycemic activity by estimating various biochemical parameters. Results: Oral administration of AC methanol extract showed significant restoration of the levels of blood glucose level. After 21 days of treatment, blood glucose, lipid profile (total cholesterol, LDL and HDL-cholesterol), glucose 6-phosphatase, fructose 1,6 bis phosphatase levels and hepatic markers enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase) were decreased when compared with diabetic control. Plasma insulin, tissue glycogen, glucose- 6-phosphate dehydrogenase levels were increased significantly compared to diabetic control. Concurrent histopathological studies of the pancreas showed comparable regeneration by extract which were earlier necrosed by STZ. Conclusions: The results exhibited that AC methanol extract possess potent antihyperglycemic activity in normal and STZ induced diabetic rats and so might be of useful in the management of diabetes. The phyto-treatment showed more efficient antihyperglycemic effect than the standard drug glibenclamide.

953-961

Authors: Raja Chakraborty, Biplab De, Nayakanti Devanna, Saikat Sen

Objective: To evaluate analgesic, anti-inflammatory and in vitro antioxidant potential and determine total phenolic, total flavonoid content of leaves extracts of Phyllanthus acidus, a folk medicinal plant of India. Methods: Anti-inflammatory activity was evaluated using carrageenan induced paw oedema, cotton pellet induced granuloma, membrane stabilizing activity method. Analgesic activity of the extracts was estimated against acetic acid induced writhing, tail immersion method, formalin test. Free radical scavenging and antioxidant potential of the extracts of Phyllanthus acidus leaves was performed using several in vitro and ex vivo assay models. Total phenolic and total flavonoid contents of the extracts were determined using standard chemical methods. Results: The extracts exhibited significant anti-inflammatory and analgesic activities at dose dependent manner. Methanol extract at a dose of 500 mg/kg showed superior activity which was comparable with the standard drugs. Ethyl acetate extract showed moderate activity while petroleum ether extract showed least activity. Total phenolic and total flavonoid content in methanol extract were 73.08±0.682 mg GAE/g and 61.28±0.062 mg QE/g respectively. The extracts possess significant antioxidant activity, methanol extract showed highest IC50 value. The contents of flavonoids and phenolic compounds could be correlated with the antioxidant, analgesic and anti-inflammatory activities observed for Phyllanthus acidus leaves. Conclusion: Our findings suggest that Phyllanthus acidus contains potential antioxidant, analgesic and antiinflammatory compounds which could be tested as drug candidates against oxidative stress, pain and inflammation related pathological diseases.

970-975

Authors: Sheetal S Chaudhari, Sanjay R Chaudhari, Machindra J Chavan

Objective: To evaluate the analgesic, anti-inflammatory and anti-arthritic activity of leaves of Cassia uniflora (C. uniflora) Mill. Methods: Petroleum ether, ethyl acetate and methanolic extract of C. uniflora (100 and 200 mg/kg, body weight) was screened for analgesic (Eddy's hot plate and acetic acid induced writhing), anti-inflammatory (Carrageenan induced paw edema) and anti-arthritic (Complete Freund's Adjuvant induced arthritis). In Complete Freund's Adjuvant arthritis model degree of inflammation was evaluated by hind paw swelling, body weight, and biochemical parameters and supported by radiological analysis. Results: Treatment with extracts of C. uniflora showed significant (P<0.05) and dose dependant increase in paw licking time in Eddy's hot plate method. In writhing test, extracts were significantly reduced the number of writhes. A dose dependant and significant inhibition of edema was observed in carrageenan induced paw edema. Petroleum ether extract at a dose of 100 mg/kg body weight showed most potent and significant activity which is supported by the results of body weight, biochemical parameters and radiological analysis in complete Freund's Adjuvant arthritis model. Conclusion: The extract possesses analgesic, anti-inflammatory and anti-arthritic activity which may be mediated through the phytochemical constituents of the plant.

981-984

Authors: Oluwatoyin Adenike FABIYI, Olubunmi ATOLANI, Oluyomi S. ADEYEMI, Gabriel Ademola OLATUNJI

Objective: The objective of this work was to determine the beta-amyrin acetate fraction in leave extract of Bridelia ferruginea and evaluate for its antioxidant and cytotoxicity potentials. Methods: The dried and pulverized leaves of Bridelia ferruginea was extracted with hexane and then with ethyl acetate. The concentrated ethylacetate extract subjected to silica gel column chromatography and eluted with a mixture of equal volume of hexane and dichloromethane afforded two major fractions. The more polar fraction was concentrated and subjected to GCMS analysis which afforded the steroid, 12-Oleanen-3yl acetate commonly known as beta-amyrin acetate (66.14%). Its ability to act as a scavenger of DPPH radical and its cytotoxicity potential based on brine shrimp assay were investigated. Results: The DPPH antioxidant assay revealed that the fraction had a higher antioxidant potential with an IC50 value of 158.2µg/mL relative to gallic acid which had IC50 of 201.1 µg/mL. The cytotoxicity assay using the brine shrimp a gave LC 50 values of 319 and 5.86 µg/mL for acute and lethal doses respectively indicating extreme toxicity when compared to reference drug, cyclophosphamide which had LC50 value of 2506 µ g/ mL. Conclusions: Thus, the beta-amyrin acetate has been identified for the first time in the leave of Bridelia ferruginea. The data here suggest that the beta-amyrin acetate fraction of the leave of Bridelia ferruginea could be further explored in biological profiling requiring antioxidant and cytotoxic dependent therapeutics as the plant could be a viable source of antioxidant and cytotoxic agents in cancer chemotherapy in the near future.

988-994

Authors: Bikram K. Mallik, Tribhuban Panda, Rabindra N. Padhy

Objective: To record ethnobotanical information from a hill-dwelling aboriginal tribe of Odisha. Methods: Gathering information on medicinal uses of plants by moving door to door for conducting personal interviews with socio-economically backward tribals (aborigine people) of Niyamagiri hill area of Kalahandi district, Odisha state, India and recording the accumulated knowledge with the snowball technique. Plants were identified by the users on forest floor and were botanically classified. Results: This communication consists of uses of parts of 111 plant species belonging to 105 genera of 59 plant families, against 68 human ailments, diarrhoea, dysentery, scabies, intestinal worms, gastrointestinal disorders, venereal disease, gynecological disorders, gingivitis, rheumatism, joint pains, wounds, cut injuries, mouth troubles, ear diseases, acute eye infections, foot inflammation, foot crack and eczema, particularly. These plants are used as herbal healing sources, as a part of cultural practice of aborigines down the ages. Among these plants, many are specific to the climate identified zone of the hilly forest patches of the district. Conclusions: The present record of ethnomedicinal data indicated that the backward local ethnic people use plants from their surroundings, as healing sources for all possible ailments.

1002-1009

Authors: Rajaretinam Rajesh Kannan, Samuel Gnana Prakash Vincent

Objective: To screen the herbal extracts influencing the hematopoietic stem cells (HSC) in zebrafish embryos and their chemical genetic effects. Methods: The herbals used in this study had been widely applicable in Siddha medicines in South India. Herbal extracts were treated in zebrafish embryos at 4 d post fertilization and the extracts inducing the HSC were enumerated in hemocytometer. The biocompatibility and the organogenesis of the screened extracts were assessed in the zebrafish embryos for their chemical genetic effects. The LC50 values were calculated with their parallel control. The blood cells were enumerated. Results: The level of RBC was found increased in the Bergera koenigii (B. koenigii) at 15 µg/mL (P<0.05), Mimosa pudica (M. pudica) at 20 µg/mL (P<0.05) and Solanum trilobatum (S. trilobatum) at 25 µg/mL (P<0.05) and decreased RBC level was found in Phyllanthus niruri (P. niruri) at 30 µg/mL (P<0.05). The WBC count was found increased in S. trilobatum at 20 µg/mL (P<0.05) and Annona muricata (Annona muricata) at 15 µg/mL (P<0.05) and the Vitis quadrangularis (V. quadrangularis) at 20 µg/mL (P<0.05) decreased the WBC level. There were no notable effects in heart beats and the chemical genetic effects were observed at higher concentration of the extract resulting in Pericardial bulging, trunk tail flexure with heart edema, fin fold deformities etc. Conclusions: This in vivo based screening of Hematopoiesis is an inexpensive assay to screen herbal compounds and found that S. trilobatum extract influenced embryonic HSC in zebrafish, which could be a therapeutic fo r blood related disorders.

1013-1016

Authors: Rasingam L, Jeeva S, Kannan D

Objective: To identify and understand the utilization and prioritization of medicinal plants used as tooth sticks by the select communities of Andaman and Nicobar islands. Methods: The information was collected through questionnaires and discussions among the informants in their local language regarding the plant parts used. Results: A total of 11 plant species belonging to 10 genera and 8 families were enumerated as tooth sticks, used by the Chota Nagpuri and Tamil inhabitants of Andaman and Nicobar islands to treat dental caries. Conclusion: The most important plant species harvested for tooth sticks belongs to the family Euphorbiaceae (3 species) and important as the tribal have used these plants since time immemorial and found effective in their teeth and gums health and this study has scopes on the conservation of certain medicinal plants, through sustainable utilization.

1024-1030

Authors: Ramaiyan Dhanapal, J.Vijaya Ratna, I. Sarathchandran, Malaya Gupta

Objective: To explore the antispermatogenic and testicular antisteroidogenic activities of Feronia limonia fruit pulp used traditionally to induce sterility in men in southern India. Methods: Fourty Wistar male albino rats (Rattus norvegicus) were equally divided into four groups. Experimental groups were administered with the ethanolic extract of Feronia limonia fruit pulp at doses of 250 and 500 mg/kg body weight once daily for 55 days. All the treated rats had corresponding recovery groups. At the end of each treatment periods, various spermatological indices, tissue biochemicals and testicular enzymes levels were analysed. Blood profiles were also estimated. Results: Compared with the control, the F. limonia fruit pulp at both dose levels did not decrease body weight, while the testes, epididymides and seminal vesicles were significantly (P<0.01) reduced this effects were associated with decline in epididymal sperm count, motility, viability and increased percent of abnormal sperm. Further, the Feronia limonia fruit pulp at 500 mg/kg body weight markedly reduced the epididymal and testicular protein content by 24.58 and 29.86%, respectively, as well as the glucose-6-phosphate dehydrogenase (G-6-PDH) and ∆5-3β-hydroxy steroid dehydrogenase (∆5-3β-HSD) levels by 42.82% and 38.08%, respectively, while a significant elevation was observed in testicular cholesterol and ascorbic acid content. A gradual recovery of all the parameters was observed after 55 days of treatment withdrawal. No significant alterations in haematological indices were observed. Conclusion: The present findings indicate that F. limonia fruit pulp may have reversible antispermatogenic and antisteroidogenic properties, and could partially support the traditional use as male contraceptive.

1036-1039

Authors: Shaikh M Mohsin Ali, Mele Jesmin, M Abul Kalam Azad, M Khairul Islam, Ronok Zahan

Objective: To study anti-inflammatory and analgesic activities in swiss albino mice, two schiff bases namely acetophenone semicarbazone (ASC) and benzophenone semicarbazone (BSC) were synthesized and characterized. Methods: Two doses of the test compounds 25 and 50 mg/kg (p.o) fo r each were selected throughout the research work. The anti-inflammatory activity of the test compounds was determined by ‘carragenan induced mice paw edema inhibition’ method. The analgesic activity was determined by both, ‘acetic acid induced writhing’ and ‘tail immersion' methods. All such data were compared with standard drugs at the dose of 10 mg/kg (p.o.). Results: Both ASC and BSC have showed positive effects as anti-inflammatory and analgesic agents. Anti-inflammatory and analgesic activities of the test compounds at 50 mg/kg (p.o.) were quite comparable to those of standard drugs at 10 mg/kg (p.o.). Conclusion: Both ASC and BSC can be considered as potent anti-inflammatory and analgesic agents.

1047-1051

Authors: Ramachandran M, Elumalai EK

Objective: To investigate the definite crystal structure of UDP-glucose 4-epimerase enzymes (EC 5.1.3.2) from Catharanthus roseus (C. roseus) for further research activities. Methods: The structure was modeled using homologous templates. The model validated under PROCHECK and WHAT-IF. Results: The model constructed using Modeller9v7 was validated. Moreover, 89% of residues lie in the most favored region. The model was checked for its grand average of hydropathicity and three binding sites were predicted using Molsoft ICM Pro v3.5. Conclusions: The model was suggested to be the good model. The constructed model can be used for further pharmacological studies and it can act as potential target against novel inhibitors.

1060-1063

Authors: Atul G. Babar, Anant Pande, Balasaheb G. Kulkarni

Objective: To identify the bioactive potential of crude methanolic extracts of five gastropods and one bivalve collected from the west coast of India (Mumbai coast). Methods: Discdiffusion assay and micro-dilution technique were used to test the bioactive potential of these molluscs against six pathogenic bacteria. Results: The crude methanolic extracts of all the molluscs showed significant activity against one or more bacteria tested. Minimum inhibitory concentration of crude methanolic extracts was found in the range of 0.10–0.35 mg/mL against human pathogenic bacterial strains. Hemifusus pugilinus and Nerita sp. extracts were most effective showing activity against all bacterial strains. Hemifusus pugilinus extract exhibited good activity against Vibrio cholerae contrary to earlier reports. Methanolic extract of Gafrarium divaricatum demonstrated considerable activity against both Escherichia coli and Streptococcus pyogenes. Trochus radiatus extract was inactive against all the bacteria tested except Escherichia coli. Euchelus asper extract was active against Klebsiella pneumoniae and Pseudomonas aeruginosa. Conclusions: All the molluscs screened exhibited potential activity against one or more bacterial strain. Further work is in progress to isolate the active constituents of the extracts.

1070-1076

Authors: Palavesam Suganthi, Sundaram Ravikumar

Objective: To investigate the crude extract of marine actinomycetes with adverse effect locally on the adult Wister albino rats or systematically in the blood circulation. Methods: Acute toxicity, sub acute toxicity, biochemical and histopathological were tested. Results: In the results acute toxicity (LD50=2 500 µg/kg bw), sub acute toxicity study (2 500 µg/kg bw) were significant at 5% level of each experimental groups compared to the control group. Biochemical and histopathological study also showed better as compared with control group Conclusion: This crude microbial extract from Streptomyces sp. RSAUT 20 and Streptomyces scabiei (S. scabiei) RSAUK 49 is potential source for novel antimicrobial compounds. The crude extract of Streptomyces sp. RSAUT 20 and S. scabiei RSAUK 49 were tested for in vivo toxicity study.

1081-1085

Authors: Alaganadham Elavarasi, Gnanaprakash Sathiya Rathna, Murugaiyan Kalaiselvam

Objective: To find out the anticancer properties of Taxol (paclitaxel) isolated from mangrove endophytic fungi. Methods: An endophytic fungus Fusarium oxysporum was isolated from Rhizophora annamalayana, a mangrove plant and analysis for Taxol production. The fungus was identified based on morphology and spore characteristics. The secondary metabolites Taxol were extracted with ethyl acetate. Taxol extracted was characterized by chromatographic and spectrometric analysis. Results: Thin layer chromatography plate shows violet red and IR spectrum values were conformed as group of terpenoid functional groups. The HPLC analysis showed the higher yield of Taxol 172.3 µg/L from potato dextrose liquid medium. Conclusion: The bioprospecting of entophytic fungus F. oxysporum isolated from mangrove is discussed and may serve as a potential material for the production of Taxol for anticancer treatment.

1090-1095

Authors: Samuel Jacob Inbaneson, Sundaram Ravikumar

Objective: To identify the possible antiplasmodial drugs from bacteria associated with marine sponge Clathria indica. Methods: Clathria indica samples were collected from Thondi coast and subjected for enumeration and isolation of associated bacteria. Filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125 µg.mL-1) from isolated bacterial isolates were screened for antiplasmodial activity against Palsmodium falciparum and potential extracts were also screened for biochemical constituents. Results: The count of bacterial strains were maximum in November 2007 (19× 104 CFU.g-1) and the average count was maximum during the monsoon season (107×103 CFU.g-1). Thirty one morphologically different bacterial isolates were isolated from Clathria indica and the ethyl acetate bacterial extracts were screened for antiplasmodial activity against Palsmodium falciparum. The antiplasmodial activity of a isolate THB23 (IC50 28.80 µg.mL-1) extract is highly comparable with the positive control chloroquine (IC50 19.59 µg.mL-1) and 17 bacterial extracts which showed IC 50 value of more than 100 µg.mL-1. Statistical analysis reveals that, significant in vitro antiplasmodial activity (P<0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes showed no morphological changes in erythrocytes by the ethyl acetate extract of bacterial strains after 48 h of incubation. The in vitro antiplasmodial activity might be due to the presence of carbohydrates and alkaloids in the ethyl acetate extracts of bacterial isolates. Conclusions: The ethyl acetate extracts of THB23 possesses novel compounds for the development of antiplasmodial drugs.

1102-1115

Authors: G.Marichamy, M.A.Badhul Haq, R.Vignesh, R.Shalini, A.R.Nazar

Objective: The objective of the study was to evaluate the essential and non essential fatty acids and the distribution of Omega-3 and Omega-6 fatty acids in twenty commonly consumed edible fi shes of parangipettai coastal waters. Methods: For fatty acid analysis, each fish specimens were beheaded, eviscerated and filleted manually. The tissue samples were oven dried at 67°C for 24hrs.After that the samples ware grounded finely with pestle and mortar. The saponified samples were cooled at room temperature for 25 min, they were acidified and methylated by adding 2 ml 54% 6 N Hcl in 46% aqueous methanol and incubated at 80°C for 10 min in water bath. Following the base wash step, the FAMEs were cleaned in anhydrous sodium sulphate and then transferred in to GC sample vial for analysis. FAMEs were separated by gas chromatograph. Results: The results of the present study revealed that the most abundant individual FAs were Palmitic acid, Oleic acid, Arachidonic acid (AA), Docosahexaenoic acid (DHA) in most the tissues. The total Arachidonic acid (C20:4ω-6) was found to be higher proportion (0.17-4.86%), when compared with other Omega-6 fatty acids. The values found for Linoleic acid (C18:2 ω-6) ranging from 0-7.23%. Siganus javus has 7.23% of Linoleic acid. Conclusion: Fatty acids are the principle components in lipids. The nutritional importance of fish consumption is in great extent associated with the content of omega-3 fatty acids. Sea food is an important dietary food for human beings. It constitute higher amount of protein, lipids, vitamins and essential and nonessential metals and low concentration of carbohydrates.

1120-1129

Authors: MA Badhul Haq, K Kalai Priya, R Rajaram, R Vignesh, M Srinivasan

Objective: To investigate the level of WSSV transmission from the infected tiger prawn Penaeus monodon (P. monodon) to specific pathogen free Litopenaeus vannamei (L. vannamei) in laboratory captivity condition in relation to PCR detection, histopathological observation and viral genome sequence. Methods: Viral DNA was isolated from purified virions by treatment with proteinase K (0.2 mg/mL) and Sarkosyl (1%). The purity and concentration of the DNA were determined by agarose gel electrophoresis. Moribund and dead shrimp were removed and processed for indirect immunofluorescence (IIF) analysis. Histological observation of infected L. vannamei shrimps were revealed by the degenerated cells which were characterized by intranuclear inclusions in the tissues of WSSV infected mid-gut gland, lymphoid organ, gill lamellae and gut epithelium. Total DNA was extracted, from shrimp hemolymph and tissues, with a High Puree PCR template preparation kit. WSSV-DNA was detected using a commercial 2-step PCR detection kit. Results: The present study compares the real-time PCR results with SYBR Green I concentration ranging fr om 0.2 to 0.7×. The positive standard was used in the range of 102, 104 106, 108 and 1010 copies/ng of DNA in general. The PCR analysis showed the appearance of a prominent band from the PCR amplified product of WSSV-DNA at internal control band of 848 bp. Moderate and severe levels were observed as 650 bp and 910 bp (200 & 2 000 copies) in various transmission routes. The WSSV content in moribund shrimp of all the experimental species (L. vannamei) approximately ranged in nucleotide application by quantification method from 0.000 001 WSSV copies/µg of total DNA. In whole moribund infection animal, approximately 0.02 WSSV copies/µg of DNA was detected in nucleotide applied animal. Conclusion: These results indicate that wild brood stock and native culture shrimp P. monodon may be infected with WSSV and can get transferred into the SPF L. vannamei farming environment. Based on the studies, we made in captivity condition in different WSSV transmission route in dissimilar infection range with the use of nucleotide for antiviral drugs. There is an urgent need to address and develop antiviral drugs and molecular based viral genome technique for control measures to salt away the aquaculture environments.

1135-1137

Authors: Faranak Firoozfar, Nahid norjah, Mojgan Baniardalani, Seyed Hassan Moosa-Kazemi

Objective: The study was carried out to determine the knowledge, believe and practice of students about insects for forthcoming research due to tropical medicine. Its application to tropical medicine is acceptable. Methods: The study was conducted among 300 students by pretrained interviewers using a pre-test questionnaire in secondary, high school, and university in the Tehran city from June 2008 to May 2009. All statistics analysis was carried out the spss ver 11.5 EPI6, stata 80, chi-square and fisher exact test. Results: Significantly, 82.3% of the interviewers had problem when expose to insects (P<0.05). Domestic animals and unhygienic human habitant most importantly incriminate to present of insects. Out of 73.8% of the respondents believed the association between entomophobia and human habitat. Entomophobia was significantly associated with density and size of insects (P<0.05). Out of 37% of the respondents agree with killing of insects pest (P<0.05). Significantly steers was the common sign of entomophobia among the respondents (P<0.05). Out of %49.7 of audiences belived to association between entomophobia and age.Conclusion: In conclusion important the education and communication messages against entomophobia take in to consideration the local practice.

1147-1150

Authors: Smriti Singh, Pramod Kumar, N Gopalan, Bhuvnesh Shrivastava, RC Kuhad, Hotam Singh Chaudhary

Objective: To isolate strains of Actinomycetes from different locations of Gwalior to evaluate its antimicrobial activity against multidrug resistant pathogenic strains. Method: Soil samples collected from different niche habitats of Gwalior were serially diluted and plated on selective media. Potential colonies were further purified and stored in agar slants and glycerol stocks. Isolates were biochemically characterized and purified isolates were test against pathogenic microorganisms for screening. Isolates with antagonistic properties were inoculated in production media and secondary metabolites or antimicrobial products were extracted. Result: The seven actinomycetes strains showing maximum antibacterial activity were isolated further characterized based on their colony characteristics and biochemical analyses. The isolates were screened for their secondary metabolites activity on three human pathogenic bacteria are Escherichia coli (E. coli), Methicillin-Resistant Staphylococcus aureus (S. aureus) and Vancomycin-Resistant Enterococci (VRE). Discussion: The strain MITS 1005 was found to be more active against the test bacteria.

1156-1158

Authors: Saraei M1, Zakilo M, Tavazoei Y, Jahanihashemi H, Shahnazi M

Objective: To define contamination of parks to Toxocara eggs in Qazvin city which is the nearest province center to Tehran, capital of Iran. Methods: A total of 180 soil and grass samples were collected from three city parks and examined for Toxocara spp. eggs by sedimentationfl otation method. Results: Overall, 5.8% of samples were positive for Toxocara spp. eggs with contamination rate of 3.15% and 8.42% for soil and grass samples (P>0.05), respectively. The fr equency of egg numbers in positive samples was between 1 to 5 and all unembryonated. Conclusions: It seems that the low frequency of pet dogs is the main cause of low contamination by Toxocara spp. eggs in this city.

1162-1168

Authors: Atanu Bhattacharjee, Shastry Chakrakodi Shashidhara, Aswathanarayana

Crataeva nurvala Buch-Hum (Varuna) is well known traditional Indian medicinal plant used to treat various ailments in particular urolithiasis. During last two decades, numerous ethnopharmacological and scientific reports have been cited in the literature to support its multidirectional therapeutic potential. The plant is rich in alkaloids, saponins, triterpenes, tannins, fl avanoid glycosides, glucosinolates and phytosterols. The review emphasizes primarily on fo lkloric uses, biological activities of isolated compounds, pharmacological activities of the extracts, clinical studies and safety profile of Crataeva nurvala to provide a comprehensive data fo r researchers to hit upon new chemical entity responsible for its claimed traditional uses.

1176-1188

Authors: Mayank Gangwar, Ragini Singh, RK Goel, Gopal Nath

Liposomes have been widely investigated since 1970 as drug carriers for improving the delivery of therapeutic agents to specific sites in the body. As a result, numerous improvements have been made to make this technology potential the treatment of certain diseases in the clinics. This review mainly focused on various aspects related to the vesicular system, including method of preparation, stabilization, drawbacks, and applications. Various types of vesicular systems such as liposomes, niosomes, transfersomes, pharmacosomes, and nanoparticle have been discussed briefly along with some other emerging vescicular systems (photosomes, archaesomes, genosomes, cryptosomes, discomes) focusing on cell specific gene transfer, photodynamic therapy and ligand mediated drug targeting. Present applications of the liposomes are in the immunology, dermatology, vaccine adjuvant, eye disorders, brain targeting, infective disease and in tumour therapy. The new developments in this field are of specific binding properties of a drug-carrying liposome to a target cell such as a tumor cell and specific molecules in the body (antibodies, proteins, peptides etc), stealth liposomes which are especially used as carriers for hydrophilic (water soluble) anticancer drugs like doxorubicin, mitoxantrone and bisphosphonate-liposome mediated depletion of macrophages. This review would help researchers working in the area of liposomal drug delivery.

1194-1199

Authors: CP Kashyap, Vikrant Arya, Narender Thakur

Traditional herbal medicines are practiced in the entire world and their ethnopharmacological records reveal that most of the people of the world have been using plants, animals, microorganisms and minerals for treating various diseases. Crataegus oxyacantha (C. oxyacantha) Linn. (Rosaceae) commonly known as Hawthorn is an official plant in Homeopathic System of Medicine to treat various conditions of cardiovascular system. In recent times, this drug has been subjected to phytochemical, pharmacological, pre-clinical and clinical investigations and many new investigations have been indicated. Current review finds ethnomedicinal and phytopharmacological potential of leaves, flowers, berries, bark etc. for exploring the immense medicinal potential of Hawthorn. At the same time, studies to evaluate the dosage, toxicity and interactions with drugs and herbs on simultaneous use, which is imperative for optimal and safe utilization of this plant, are explained. There are few comprehensive reports available on clinical use of Hawthorn in chronic heart failure patients have shown promising results.

1206-1208

Authors: Ali Raghibi, Wan Hazabbah Wan Hitam, Raja Azmi Mohd Noor, Zunaina Embong

A 58-year-old Malay lady, presented with gradual loss of vision in the right eye for 1 month duration. It was associated with right sided headache, nausea and vomiting. Patient had a history of contact with tuberculosis patient. On examination, visual acuity in right eye was no perception of light with presence of relative afferent pupillary defect. The left eye visual acuity was 6.0/7.5. There was a total restriction of extraocular movement in all direction (frozen eye) in the right eye. Both anterior segments were unremarkable. Fundoscopy in both eyes showed normal appearance of optic disc and retinal vessels. Other cranial nerves were normal. Lungs were clear and had no lymphadenopathy. Chest radiograph was normal. Erythrocyte sedimentation rate was elevated by 35 mm/h and Mantoux test was positive (20 mm). MRI of brain and orbit demonstrated thickening and irregularity of the right optic nerve on axial view with doughnut sign on coronal view. Patient was diagnosed to have optic perineuritis secondary to tuberculosis. She was treated with anti-TB drugs. Systemic corticosteroid was commenced after 2 weeks. The visual acuity was improved to hand motion after 1 month with almost complete resolution of extraocular movement.

1–5

Authors: Wesely Edward Gnanaraj, Johnson Marimuthu Antonisamy, Mohanamathi RB, Kavitha Marappampalyam Subramanian

Objective To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants. Methods Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3–4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration. Results Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition, Sixty five percentages of plants were established in the field. Conclusions The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A. aspera and A. bidentata. The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can be easily adopted for commercial large scale cultivation.

11–15

Authors: M Nirmala, K Girija, K Lakshman, T Divya

Objective To investigate the hepatoprotective activity of stem of Musa paradisiaca (M. paradisiaca) in CCl4 and paracetamol induced hepatotoxicity models in rats. Methods Hepatoprotective activity of alcoholic and aqueous extracts of stem of M. paradisiaca was demonstrated by using two experimentally induced hepatotoxicity models. Results Administration of hepatotoxins (CCl4 and paracetamol) showed significant biochemical and histological deteriorations in the liver of experimental animals. Pretreatment with alcoholic extract (500 mg/kg), more significantly and to a lesser extent the alcoholic extract (250 mg/kg) and aqueous extract (500 mg/kg), reduced the elevated levels of the serum enzymes like serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP) and bilirubin levels and alcoholic and aqueous extracts reversed the hepatic damage towards the normal, which further evidenced the hepatoprotective activity of stem of M. paradisiaca. Conclusions The alcoholic extract at doses of 250 and 500 mg/kg, p.o. and aqueous extract at a dose of 500 mg/kg, p.o. of stem of M. paradisiaca have significant effect on the liver of CCl4 and paracetamol induced hepatotoxicity animal models.

21–23

Authors: T Thirumalai, E David, Therasa S Viviyan, EK Elumalai

Objective To evaluate the effect of aqueous seed extract of Solanum surattense (S. surattense) on the oxidative potential of cauda epididymal spermatozoa. Methods S. surattense seed extract was orally administered at the dosage of 10 mg/kg b.w. for 15 days, after which aspartate transferase (AST), alanine transferase (ALT), glutamate dehydrogenase (GDH), citric acid and iso-citrate dehydrogenase (ICDH) were assayed. Results The activity levels of the enzymes AST and ALT, which are considered to be the androgenicity in the sperm suspension, were depleted in the extract fed rats. The activity level of the enzyme ICDH, was reduced significantly in the treated group (P<0.001). Conclusions It can be concluded that the oral administration of the aqueous seed extract of S. surattense can deplete the oxidative stress of cauda epididymal spermatozoa in albino rats.

31–35

Authors: Prabu K Ananda, CT Kumarappan, Christudas Sunil, VK Kalaichelvan

Objective To investigate the effect of aqueous solution of Biophytum sensitivum leaf extract (BSEt) on normal and streptozotocin (STZ)-nicotinamide-induced diabetic rats. Methods Diabetes was induced in adult male Wistar rats by the administration of STZ-nicotinamide (40, 110 mg/kg b.w., respectively) intraperitoneally. BSEt (200 mg/kg) was administered to diabetic rats for 28 days. The effect of extract on blood glucose, plasma insulin, total haemoglobin, glycosylated haemoglobin, liver glycogen and carbohydrate metabolism regulating enzymes of liver was studied in diabetic rats. Results BSEt significantly reduced the blood glucose and glycosylated haemoglobin levels and significantly increased the total haemoglobin, plasma insulin and liver glycogen levels in diabetic rats. It also increased the hexokinase activity and decreased glucose-6-phosphatase, fructose-1, 6-bisphosphatase activities in diabetic rats. Conclusions The results of our study suggest that BSEt possesses a promising effect on STZ-nicotinamide-induced diabetes.

41–46

Authors: Nabil M Ibrahim, Esam A Eweis, Hossam S El-Beltagi, Yasmin E Abdel-Mobdy

Objective To evaluate the effect of different doses of lead acetate (1/20, 1/40 and 1/60 of LD50) on body weight gain, blood picture, plasma protein profile and the function of liver, kidney and thyroid gland. Methods Male albino rats were divided into four groups, the first group represented the health control animals, while the second, third and fourth groups were ingested orally with sub lethal doses of lead acetate (1/20, 1/40 and 1/60) of the oral LD50, respectively. One dose was ingested every two days during the experimental period (14 weeks) including the adaptation time. Blood was collected and used for all analysis. Results The results showed that, the ingestion of Pb2+ induced significant stimulation in glutamic-pyruvic transaminase (ALT) and glutamic-oxalacetic transaminease (AST) activity. Also, total soluble protein and albumin contents of plasma were significantly decreased, while the content of globulin was changed by the Pb2+ treatments. The cholinesterase activity was inhibited, but the activities of alkaline and acid phosphates and lactate dehydrogenase were stimulated, while plasma glucose level was elevated as a result of lead acetate intoxication. In case of blood picture, Pb2+ ingestion reduced the contents of hemoglobin and RBCs count of intoxicated rat's blood and the plasma levels of T3, T4 and blood WBCs count were decreased. Conclusions It can be concluded that lead acetate has harmful effect on experimental male albino rats. Therefore, the present work advises people to prevent exposure to the lead compound to avoid injurious hazard risk.

50–54

Authors: Zailatul Hani Mohamad Yadzir, Rosmilah Misnan, Noormalin Abdullah, Faizal Bakhtiar, Masita Arip, Shahnaz Murad

Objective To characterize the major allergens of Macrobrachium rosenbergii (giant freshwater prawn). Methods Raw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools. Results SDS-PAGE of the raw extract showed 23 protein bands (15–250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients' sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase. Conclusions It can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies.

61–65

Authors: Tan Zi Ning, Wong Weng Kin, Shaymoli Mustafa, Arefuddin Ahmed, Rahmah Noordin, Tan Gim Cheong, Olivos-Garcia Alfonso, Lim Boon Huat

Objective To compare the efficacy of three different tissue stains, namely haematoxylin and eosin (H&E), periodic-acid Schiff (PAS) and immunohistochemical (IHC) stains for detection of Entamoeba histolytica (E. histolytica) trophozoites in abscessed liver tissues of hamster. Methods Amoebic liver abscess was experimentally induced in a hamster by injecting 1 × 106 of axenically cultured virulent E. histolytica trophozoites (HM1-IMSS strain) into the portal vein. After a week post-inoculation, the hamster was sacrificed and the liver tissue sections were stained with H&E, PAS and IHC stains to detect the amoebic trophozoite. Results The three stains revealed tissue necrosis and amoebic trophozoites, but with varying clarity. H&E and PAS stained the trophozoites pink and magenta, respectively, however it was difficult to differentiate the stained trophozoites from the macrophages because of their similarity in size and morphology. On the other hand, IHC stain revealed distinct brown appearance of the trophozoites in the infected liver tissues. Conclusions It can be concluded that out of the three stains, IHC is the best for identification of E. histolytica trophozoites in tissue sections.

70–75

Authors: Yan-Bo Zeng, Shun-Hai Zhu, Hui Dong, Hong-Yu Han, Lian-Lian Jiang, Quan Wang, Jun Cheng, Qi-Ping Zhao, Wei-Jiao Ma, Bing Huang

Objective To identify more effective and less toxic drugs to treat animal toxoplasmosis. Methods Efficacy of seven kinds of sulfonamides against Toxoplasma gondii (T. gondii) in an acute murine model was evaluated. The mice used throughout the study were randomly assigned to many groups (10 mice each), which either remained uninfected or were infected intraperitoneally with tachyzoites of T. gondii (strains RH and CN). All groups were then treated with different sulfonamides and the optimal treatment protocol was determined candidates. Sulfadiazine-sodium (SD) was used for comparison. Results The optimal therapy involved gavaging mice twice per day with 250 mg/kg bw of sulfachloropyrazine-sodium (SPZ) for five days. Using this protocol, the average survival time and the time-point of 50% fatalities were prolonged significantly compared with SD treatment. Treatment with SPZ protected 40% of mice from death, and the heart and kidney tissue of these animals was parasite-free, as determined by nested-PCR. SPZ showed excellent therapeutic effects in the treatment of T. gondii in an acute murine model and is therefore a promising drug candidate for the treatment and prevention of T. gondii in animals. Conclusions It can be concluded that the effective drug sulfachloropyrazine may be the new therapeutic options against animal toxoplasmosis.

80–82

Authors: Bava Javier, Lloveras Susana, Garro Santiago, Troncoso Alcides

We communicate the diagnosis by microscopy of a pulmonary coinfection produced by Cryptococcus neoformans and Pneumocystis jiroveci, from a respiratory secretion obtained by bronchoalveolar lavage of an AIDS patient. Our review of literature identified this coinfection as unusual presentation. Opportunistic infections associated with HIV infection are increasingly recognized. It may occur at an early stage of HIV-infection. Whereas concurrent opportunistic infections may occur, coexisting Pneumocystis jiroveci pneumonia (PCP) and disseminated cryptococcosis with cryptococcal pneumonia is uncommon. The lungs of individuals infected with HIV are often affected by opportunistic infections and tumours and over two-thirds of patients have at least one respiratory episode during the course of their disease. Pneumonia is the leading HIV-associated infection. We present the case of a man who presented dual Pneumocystis jiroveci and cryptococcal pneumonia in a patient with HIV. Definitive diagnosis of PCP and Cryptococcus requires demonstration of these organisms in pulmonary tissues or fluid. In patients with < 200/microliter CD4-lymphocytes, a bronchoalveolar lavage should be performed. This patient was successfully treated with amphotericin B and trimethoprim sulfamethoxazole. After 1 week the patient showed clinical and radiologic improvement and was discharged 3 weeks later.

1-6

Authors: MD Nazneen Bobby, Wesely EG, Johnson M

Objective: To establish alkaloids profile for the medicinally important plant Albizia lebbeck (A. lebbeck) using high performance thin layer chromatography (HPTLC). Methods: Preliminary phytochemical screening was carried out by Harborne method. HPTLC studies were followed by Harborne and Wagner et al method. Ethyl acetate-methanol-water (100:13.5:10) was employed as mobile phase for alkaloids. The developed plate was sprayed with Dragendorff’s reagent followed by 10% sodium nitrite reagent as spray reagent and dried at 100 ℃ in hot air oven for 3 min. The plate was photo-documented at UV 366 nm and daylight using photo-documentation chamber. Results: Alkaloids, steroids, terpenoids, flavonoids, saponins and glycosides present in the petroleum ether, ethyl acetate and methanolic extracts of A. lebbeck leaves. The petroleum ether extracts of A. lebbeck leaves displayed the presence of 10 types of alkaloids with 10 different Rf values ranging from 0.02 to 0.85. The ethyl acetate extract of A. lebbeck leaves illustrated the presence of 5 different types of alkaloids with 5 different Rf values ranging from 0.09 to 0.84. The methanolic extract of A. lebbeck leaves demonstrated the presence of 4 different types of alkaloids with 4 different Rf values with range from 0.02 to 0.79. Maximum number (10) of alkaloids has been observed in petroleum ether followed by ethyl acetate (5) extracts of the leaves of A. lebbeck. Conclusions: In the present study we observed various alkaloids profile of the medicinally important plant using HPTLC. This profile can be used for the identification of the medicinally important plants from the adulterant. Further, separation and characterization of the bioactive compound (principles) from the plants is to be evaluated and reported in near future.

12-14

Authors: Sharad Srivastava, Adarsh Pratap Singh, Ajay Kumar Singh Rawat

Objective: To develop a simple precise and novel method for the identification and quantification of bioactive molecules ferulic acid in Lycopodium clavatum. Methods: Bioactive molecules ferulic acid in Lycopodium clavatum was indentified and estimated by the assay combined separation and quantitative estimation of the analyte on silica gel 60F254 HPTLC plates with visualization under UV and scanning at 320 nm. Results: HPTLC studies showed the separation and determination of ferulic acid (0.443%) in Lycopodium clavatum. The result of recovery ranged in 95%-97% for ferulic acid. Conclusions: This method is simple, sensitive, economic, novel and first time reported from this species. Therefore, ferulic acid might be a useful chemotaxonomic marker for the genus Lycopodium and other lycopods.

21-26

Authors: M. Johnson

Objective: To analyze the similarity and variation among different populations of Ocimum sanctum (O. sanctum) by isoperoxidase, isoesterase, acid phosphatase and alkaline phosphatase profile. Methods: The isolation, separations and staining of the enzymes isoperoxidase, isoesterase, acid phosphatase and alkaline phosphatase were performed as described by Sadasivam and Manickam. Results: In isoperoxidase system, the band PRX4 1 (0.36) was present only in the accession collected from Salem, similar to that band PRX4 2 showed its presence only in the accession collected from Kolli Hills and PRX5 1 (0.42) was expressed only in Wyanad accession. The accession collected from Tirunelveli showed its unique banding profile in region ACP2 3 with 0.18 molecular weight-Rf values in the acid phospahatase system. In esterase system, EST5 1 and 8 1 were observed only in the accession collected from Tenkasi. In the alkaline phosphatase enzyme system, only one region (AKP6) of activity with single band (AKP6 1 -0.51) was observed. In this enzyme system, all the selected accession showed their expression. Highest percentage (0.666 7) of similarity was observed between the accessions collected from Tirunelveli and Kolli Hills, Nagercoil and Kolli Hills and Salem and Andaman while the highest percentage (0.800 0) of variation was observed between the accessions collected from Salem and Andaman. The isozyme profile based cladogram showed two clusters, cluster 1 (C1) included the accessions collected from Tenkasi, Tirunelveli, Nagercoil and Kolli Hills and they showed highest percentage of divergence with the other three accessions. The cluster 2 (C2) included the accessions collected from Wyanad, Salem and Andaman. Conclusions: Unique banding profiles of esterase, peroxidase, acid phosphatase and alkaline phospahatase were observed in O. sanctum, which represent the fingerprint of medicinally important plant. Such finger printing is useful in differentiating the species and act as biochemical markers for these species in plant systematic studies in the near future.

30-33

Authors: Solomon Jeeva, Johnson Marimuthu @ Antonisamy, Cosman Domettila, Babu Anantham, Mony Mahesh

Objective: To explore the phytochemical constituents of Ulva reticulata (U. reticulata) and Sargassum wightii (S. wightii). Methods: The preliminary phytochemical screening was performed by Harborne method. Results: The results of the phytochemical screening revealed the presence of steroids, phenolic groups, saponins, tannin, flavonoids, carbohydrates, coumarins, and xantoproteins in the extracts of U. reticulata and steroids, phenolic groups, saponins, tannin, flavonoids, carbohydrates, carboxylic acid, coumarins, and xantoproteins were detected in the extracts of S. wightii. Conclusions: The solvent extracts of U. reticulata and S. wightii show a number of metabolites presence, further work will emphasize the isolation and characterization of active principles responsible for bio-efficacy and bioactivity.

40-45

Authors: Muraleedharannair Jalajakumari Mithraja, Johnson Marimuthu @ Antonisamy, Mony Mahesh， Zachariah Miller Paul, Solomon Jeeva

Objective: To examine the phyto-constituents of Adiantum caudataum (A. caudataum), Adiantum latifolium (A. latifolium), Adiantum lunulatum (A. lunulatum), Christella dentata (C. dentate) and Christella parasitica (C. parasitica), to provide chemical marker and find inter- specific variation between the medicinally important genuses. Methods: The dried and powered leaves materials (50 g) were extracted successively with 250 mL of petroleum ether, ethyl acetate, methanol, chloroform, acetone, benzene and water by using Soxhlet extractor for 8 h at a temperature not exceeding the boiling point of the solvent. The aqueous extracts were filtered using Whattman filter paper (No.1) and then concentrated in vacuum at 40°C using Rotary evaporator. The residues obtained were stored in a freezer -70 °C until further tests. Phytochemical screening of the extracts was carried out according to the standard methods. Results: A total of five plants and 30 extracts were examined for the phytochemical screening. The crude extracts of A. caudataum, A. latifolium, A. lunulatum, C. dentata and C. parasitica showed varied degree of phyto-constituents with reference to solvents of the plant extracts. A total of three plants and 18 extracts were examined for the phytochemical screening A. caudataum, A. latifolium, A. lunulatum. The steroid is totally absent in the tested extracts of A. lunulatum. Similar to that caoboxylic acid also absent in A. caudatum and Coumarin showed its presence only in A. caudatum. A total of two plants and twelve extracts were examined for the phytochemical properties of Christella. Of which phenol is present in 11 extracts tested. Steroid is present only in one extract of C. parasitica and coumarin is present only in C. parasitica five extracts. Conclusions: The present study result provids a simple and cheap tool (Chemical marker) for the characterization and identification of the medicinally important taxa. With the help the phytochemical screenings we can find inter- specific variation between the medicinally important genuses and easily we can differentiate the Adiantum and Christella genuses.

50-56

Authors: J.K. Patra, S. Gouda, S.K. Sahoo, H.N. Thatoi

Objective: To separate the bioactive compounds from crude methanol extract of leaf of Excoecaria agallocha (E. agallocha), a mangrove plant from Bhitarkanika, Odisha, India. Methods: Chromatographic separation was carried out by paper, thin layer chromatography (TLC) and column chromatographic techniques. Bioautography of the column fractionated extract and TLC chromatogram were evaluated in vitro for antibacterial and DPPH free radical scavenging activity. The HPLC and 1 H NMR spectroscopy of the extract was also carried out to detect the presence of compounds and possible functional groups in the crude plant extract. Results: Evaluation of solvent system for chromatographic separation revealed that n-butanol: acetic acid: water in the ratio of 4:1:1 was the most appropriate for the separation of phenolic compounds in the crude extract. The antibacterial bioautography screening of the TLC fractionated extract of E. agallocha showed positive activity against Staphylococcus aureus (MTCC-1141) and Pseudomonas aeruginosa (MTCC 1034). Evaluation of antioxidant properties of TLC fraction of E. agallocha showed 23.36% scavenging activity using DPPH assay at 40 µL concentration. HPLC chromatogram of the plant extract produced seven peaks for the leaf and two peaks for the stem/bark with the highest peaks at 2.563 and 2.493 minutes of retention time, respectively. The 1 H NMR spectroscopic analysis of the plant extract revealed the presence of acyclic aliphatic and α- mono substituted aliphatic group of compounds in the sample. Conclusions: Thus, it is apparent from the present study that methanol crude extract of E. agallocha is rich in aliphatic group of compounds which can be purified and characterized for possible pharmaceutical utilization.

64-66

Authors: Selvamony Sukumaran, Mony Mahesh, Solomon Jeeva

Objective: To identify the phytoconstituents present in the fronds of Tectaria zeylanica. Methods: Screening of phytochemicals was done by using the standard methods given by Harborne. Results: Phytochemical screening of the fronds of Tectaria zeylanica confirmed the presence of various derivatives such as phenols, saponins, steroids, tannins, xanthoproteins, coumarins and carbohydrates. Conclusions: The present observations suggest a possibility to establish high yielding plantlets by in vitro culture for the production of medicinally important bioactive compounds against human pathogenic microorganisms.

74-78

Authors: R. Subramanian, S. Gayathri, C. Rathnavel, V. Raj

Objective: To determine sodium (Na), iron (Fe), magnesium (Mg), manganese (Mn), lead (Pb), zinc (Zn), cadmium (Cd) and copper (Cu) in some medicinal plants regularly used in cooking in Indian curries. Methods: All the nutrients were determined by using atomic absorption spectrophotometer (AAS). Results: The average concentrations detected were ranged from 150.22 to 521.98, 782.42 to 813.84, 15.75 to 532.72, 85.72 to 101.50, 0.478 to 9.890, 0.684 to 2.751, and 11.51 to 94.05, 6.94 to 49.76 mg/kg for Na, Fe, Mg, Mn, Pb, Zn, Cd and Cu, respectively. Conclusions: The mineral and metal contents in the samples were found at different levels. Therefore, these medicinal plants are rich in some essential minerals, especially Fe and Mg which are essential for human health.

83-89

Authors: PN Ansil, SP Prabha, A Nitha, PJ Wills, V Jazaira, MS Latha

Objective: To evaluate the curative effect of the methanolic extract of Amorphophallus campanulatus (A. campanulatus) tuber (ACME) against thioacetamide induced oxidative stress in rats. Methods: The curative potential of two different doses of ACME (125 and 250 mg/kg; p.o.) was evaluated against thioacetamide (TAA) induced oxidative stress in male Wistar rats. Single dose of TAA (100 mg/kg; s.c.) was administered to the rats in all groups except the normal control. In treatment groups, rats were administered with silymarin-the standard drug (100 mg/kg; p.o.) and ACME 2, 24 and 48 h after TAA intoxication. Hepatotoxicity was assessed by quantifying the serum levels of AST, ALT, ALP and LDH. The antioxidant potential of ACME were also evaluated by the estimation of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), reduced glutathione (GSH) and lipid peroxidation (Thiobarbituric Acid Reactive Substances-TBARS) in hepatic and renal tissues. In addition, the liver sections were also evaluated for its histopathologic changes. Results: ACME significantly (P<0.05) reversed the elevation of serum AST, ALT, ALP, LDH and tissue malondialdehyde levels. Hepatic and renal GSH, GST, GR, GPx and catalase levels were remarkably increased by the treatment with the extract. Quantification of histopathological changes also supported the dose dependent curative effects of ACME. Conclusion: This study demonstrates the antioxidant and hepatoprotective activity of ACME and thus scientifically supports the use of this tuber in traditional medicine for the treatment of liver disorders.

99-103

Authors: Alak Kanti Dutta, Partha Sarathi Gope, Subrata Banik, Sukh Makhnoon, Muhammad Ali Siddiquee, Yearul Kabir

Objective: To study total phenolic content and antioxidant properties of 80% methanol extracts of ten high yielding rice varieties, five each from two different seasons namely aman and boro of Bangladesh. Methods: Total phenolic content was measured by Folin-Ciocalteau method while 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, hydroxyl ion scavenging, ferric reducing antioxidant power (Ferric reducing antioxidant power), and total antioxidant capacity (TAC) by ammonium molybdate, were used to analyze their Antioxidant properties. Results: Rice variaty BR5 of aman and BRRI dhan28 of boro season comparatively showed higher TPC and Antioxidant properties than the other rice varieties. BR22 of aman season showed the highest hydroxyl ion scavenging activity although it displayed the lowest TPC. Except for hydroxyl ion scavenging activity, aman rice varieties displayed comparatively higher total phenolic content and antioxidant property than the boro rice varieties. Conclusions: The results of the present study implies that the selected rice varieties possess moderate antioxidant capacity and therefore, can be considered as health supplements and nutraceuticals foods as rice is the staple food of Bangladesh.

112-115

Authors: S. Sundarammal, R.Thirugnanasampandan, M. Tamil Selvi

Objective: Orthosiphon thymiflorus (Roth) Sleesen (O. thymiflorus) is an aromatic plant widely distributed in the Western Ghats of South India. The present study investigates the chemical composition and antioxidant activities of O. thymiflorus essential oil isolated from the leaves. Methods: The essential oil was hydrodistilled from the leaves by using Clevenger apparatus. Isolated essential oil was chemically analyzed by gas chromatography (GC) and gas chromatography- mass spectrometry (GC-MS). In vitro antioxidant activity of the isolated essential oil was determined. Results: Chemical composition analysis of essential oil reported to the presence of thirty three compounds. The major compound was 2-isopropyl-5-methyl-9-methylene-bicyclo-1-decene (4.4.0) (42.62%) followed by carotol (16.48%), α -cadinol (6.14%) and δ-3-carene (5.15%). In vitro antioxidant activity of the isolated oil showed dose dependent free radical scavenging activity against DPPH (IC50 437.12±0.02 µg/mL) and hydroxyl radicals (IC50317.88依0.02µg/mL). Metal chelating and prevention of deoxyribose degradation activities of the oil was observed with the IC50 of 339.78±0.01µg/mL and 158.33±0.02µg/mL respectively. Antioxidant activity of the oil was low when compared with the positive controls such as BHT and ascorbic acid. Conclusion: The present study shows various chemical components present in the O. thymiflorus essential oil and with it’s significant antioxidant activity thus has great potential to be used as natural food supplement.

120-126

Authors: Prabhjit Kaur, Manish Kumar, Bikram Singh, Subodh Kumar, Satwinderjeet Kaur

Objective: To evaluate antigenotoxic, antioxidant and COX-2 inhibitory potential of ‘GG-1’ fraction isolated by Medium Pressure Liquid Chromatography (MPLC) from ethyl acetate extract of Glycyrrhiza glabra (G. glabra) L rhizomes. Methods: The antigenotoxic activity was tested in human blood lymphocytes using comet assay and SOS chromotest using PQ37 strain of Escherichia coli (E. coli) against the diagnostic mutagens viz. 4-NQO and H2O2. Antioxidant activity was determined by DPPH radical scavenging assay. It was also investigated for the COX-2 inhibitory activity using Cayman COX (ovine) inhibitory screening assay. Results: The spectroscopic data of ‘GG-1’ fraction revealed it to be 7-hydroxycoumarin (Umbelliferone). In SOS chromotest, at the concentration of 616.75 µM, umbelliferone exhibited moderate response by reducing the induction factor of hydrogen peroxide by 68.99% (IC50 223.44 µM) and that of 4NQO by 59.71% (IC50 280.74 µM). In comet assay, exhibited a good activity by inhibiting the genotoxicity of both hydrogen peroxide and 4NQO by 61.64% (IC50 330.02 µM) and 50.66% (IC50 577.83 µM) respectively. The umbelliferone exhibited free radical scavenging potential of 43.88% at 616.75 µM. Umbelliferone possessed 95.68% inhibition of COX-2 at 10 µM concentration with IC50<1. Conclusions: The antigenotoxic, antioxidant and COX-2 inhibitory properties showed by the umbelliferone suggests that it may have several applications in nutraceuticals and human health care.

131-135

Authors: Muraleedharannair Jalajakumari Mithraja, Varaprasadham Irudayaraj, Solomon Kiruba, Solomon Jeeva

Objective: To make scientific validation of the ethnomedicinal fern Drynaria quercifolia (L.) J. Smith in relation to urinary disorders. Methods: Antibacterial study was carried out on clinically isolated Urinary Tract Infecting (UTI) bacteria by disc diffusion method. Results: Among the six extracts tested against eight different UTI bacteria, acetone extract was effective against Enterococcus faecalis and Streptococcus pyogenes, while ethanol extract was effective against Pseudomonas aeruginosa. Preliminary phytochemical screening shows the presence of phytoconstituents viz. phenols, tannin, alkaloids, proteins, xanthoproteins, carboxylic acid, coumarins and saponins. Conclusions: Based on the present study, along with previous studies, the ethnomedicinal use of Drynaria quercifolia for the treatment of urinary infection has been scientifically validated.

140-146

Authors: Xavier Devanya Rosaline, Shanmugavel Sakthivelkumar, Kuppu Rajendran, Sundaram Janarthanan

Objective: To screen the antibacterial efficacy of various solvent extracts of marine algae such as Sargassum wightii (S. wightii), Chaetomorpha linum (C. linum) and Padina gymnospora (P. gymnospora) against some selected gram-positive and gram-negative human pathogenic bacteria. Methods: Crude extracts were prepared from the selected marine algae using different solvents namely, hexane, ethyl acetate, acetone and methanol and were tested for their antibacterial activity against human pathogenic bacteria using disc diffusion method. Minimum inhibitory concentration (MIC) was also performed for selected solvent extracts for all the bacterial species. A suitable positive control was also maintained. Results: Among the three marine algae screened P. gymnospora and S. wightii were found to be more active than C. linum. It was observed that the acetone extracts of all the three marine algae showed higher inhibitory activity for the selected bacterial species than other solvent extracts. The results revealed that the crude acetone extracts seem to be a good source material in identifying the effective pure antibacterial compound(s) in all the three marine algae and particularly, S. wightii. Conclusions: The present study showed that the acetone extracts of marine algae such as S. wightii, C. linum and P. gymnospora exhibited good antimicrobial activity. But the acetone extracts of S. wightii possessed highest antibacterial activity than others and so it could be useful in seeking active principles against human pathogenic bacteria.

151-154

Authors: Solomon Jeeva, Johnson Marimuthu @ Antonisamy

Objective: To investigate the antibacterial activity and phytochemical properties of Begonia floccifera Bedd. (B. floccifera) methanolic flower extracts against the selected pathogens. Methods: 20 g of fresh flowers were soaked in 100 mL methanol for 48 h at room temperature. After 48 h, the extracts were filtered using Whatman No. 41 filter paper. The filtrates were collected, made up to known volume and stored in refrigerator at 4 曟. The methanolic extracts were used for phytochemical and antibacterial studies. The preliminary phytochemical screening was performed according to the modified Harborne method. The methanolic flower extracts of B. floccifera Bedd. was tested against Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Salmonella typhi, Serratia marcescens, Proteus mirabilis, Enterococcus faecalis and Streptococcus pyogenes by the disc diffusion method. Results: The results of the phytochemical screening revealed that phenol, tannins, xanthoproteins, steroids, tannins, steroids, phytosterols, triterpenoids, sapogenins, coumarins and carbohydrates presence in the methanolic extracts of B. floccifera. The antibacterial activity has been observed in the methanolic extracts of B. floccifera against the tested bacteria with varied activity. The maximum zone of inhibition was 28 mm for Bacillus cereus, 25 mm for Staphylococcus aureus, 15 mm for Escherichia coli, 13 mm for Proteus mirabilis, 7 mm for Klebsiella pneumonia. Conclusions: The antimicrobial activity of B. floccifera methanolic flower extracts are comparable and their potential as alternative in the treatment of infectious by these microorganisms.

159-161

Authors: Sunil Christudas, Kulathivel TM, Agastian P

Objective: To investigate the phytochemical screening and antibacterial activity of pet. ether, chloroform and ethanolic extracts of Tridax procumbens (T. procumbens). Methods: The antibacterial activity of leaves of T. procumbens was evaluated by agar well diffusion method against three selected gram positive and gram negative bacterial species. Results: The presence of carbohydrates, proteins, tannins, steroids, alkaloids and purines in the different leaf extracts were established by phytochemical analysis. The chloroform extract was more effective against bacteria and the activity was comparable with that of standard drug ampicillin. Conclusions: The results in the present study suggest that T. procumbens leaves can be used for treating diseases caused by the tested organisms.

165-169

Authors: Patel DK, Prasad SK, Sairam K, Hemalatha S

Objective: To evaluate active aldose reductase (AR) inhibitory principles from ethanolic extract and one isolated compound beta sitostirol from Hybanthus enneaspermus (H. enneaspermus) Linn F. Muell. (Violaceae), used in the treatment of various complications including diabetes. Methods: AR inhibitory activities of H. enneaspermus and isolated compound beta sitosterol against rat lens AR enzyme were investigated in vitro using quercetin as a standard drug. Results: There was a significant difference in the AR inhibitory activity, and it was found to be more in H. enneaspermus [IC50 (49.46±2.26 µg/mL)] followed by beta sitosterol [IC50 (82.71±0.42 µg/mL)] but was less compared to standard quercetin [IC50 (3.326±0.11 µg/mL)]. From the value of Vmax, Km and Ki it was found that H. enneaspermus inhibited enzyme in a non competitive manner, whereas beta sitosterol inhibited enzyme in a competitive manner. Conclusions: H. enneaspermus showed significant AR inhibitory activity compared to the isolated compound beta sitosterol, which may be due to its high phenolic and flavonoid content. So this plant can be used to treat diabetic complications in the future, but further evaluation is necessary to know the exact mode of action involved in this process.

175-177

Authors: Sunder singh, Rai AK, Praveen Sharma, Yogesh Barshiliya, Mohini Sihare, Anju Negi

Objective: To evaluate the antidiarrhoeal effect of alcoholic extract of Rotula aquatica by using different experimental models. Methods: Antidiarrhoeal effect was evaluated by castor oil induced diarrhea, charcoal meal test and PGE2 induced diarrhea. Loperamide (2 mg/kg) and atropine (0.1mg/kg) were used as standard drugs. Alcoholic extract was used in 100 and 200 mg/kg dose. Results: In case of castor oil induce diarrhea dose 200 mg/kg showed delay in defecation time (186.980±15.660) min and 100 mg/kg showed delay in defecation time (97.779±4.952) min, while loperamide (2 mg/kg) showed delay in defecation time (221.148±21.919) min. Finally the reading showed that doses 200 mg/kg showed significant antidiarrhoeal effect from 100 mg/kg of alcoholic extract of Rotula aquatica in different model used for antidiarrhoeal activity. Conclusions: This study revealed that the alcoholic extracts has potential pharmacological activity against diarrhea and this may possible explain the use of the plant in traditional medicine.

181-186

Authors: Sheetal S. Chaudhari, Sanjay R. Chaudhari, Machindra J. Chavan

Objective: To evaluate the analgesic, anti-inflammatory and anti-arthritic activity of leaves of Cassia uniflora Mill. Methods: Petroleum ether, ethyl acetate and methanolic extract of Cassia uniflora (100 and 200 mg/kg, body weight) was screened for analgesic (Eddy’s hot plate and acetic acid induced writhing), anti-inflammatory (Carrageenan induced paw edema) and anti-arthritic (Complete Freund’s adjuvant induced arthritis). In complete Freund’s adjuvant arthritis model degree of inflammation was evaluated by hind paw swelling, body weight, and biochemical parameters and supported by radiological analysis. Results: Treatment with extracts of Cassia uniflora showed significant (P<0.05) and dose dependant increase in paw licking time in Eddy’s hot plate method. In writhing test, extracts significantly reduced the number of writhes. A dose dependant and significant inhibition of edema was observed in carrageenan induced paw edema. Petroleum ether extract at a dose of 100 mg/kg body weight showed most potent and significant activity which was supported by the results of body weight, biochemical parameters and radiological analysis in complete Freund’s adjuvant arthritis model. Conclusions: The extract possesses analgesic, anti-inflammatory and anti-arthritic activity which may be mediated through the phytochemical constituents of the plant.

192-196

Authors: Maharudra S Rakh, Amol N Khedkar, Nilesh N Aghav, Sanjay R Chaudhari

Objective: To evaluate the antiallergic and analgesic activities of seed of Momordica dioica Roxb. (M. dioica) Methods: Petroleum ether, ethyl acetate, methanol and aqueous extracts of M. dioica seed were screened for anti-allergic activity (200 mg/kg (i.p.) in mice body wt.) and analgesic (dose dependant 50 mg/kg and 100 mg/kg in mice body wt.). Results: Methanol extract of M. dioica seed shows more significant antiallergic activity (P<0.01) as compare to other extracts in all three models of milk induced leukocytosis, milk induced eosinophilia and differentia leukocytes count in mice. In dose dependant analgesic activity petroleum ether and methanol extracts gives significant hot plate activity (P<0.05, P<0.01) and in acetic acid induced writhing test significantly (P<0.01) reduced the number of writhing. Conclusions: The antiallergic activity may be due to phytochemical constituents such as polar constituents and flavonoids. The extract possesses analgesic activity which may be due to the phytochemical constituents of the plant.

200-202

Authors: Pallara J Wills, Velikkakathu V Asha

Objective: To investigate the acute and subacute toxicity of Lygodium flexuosum (L. flexuosum) extracts to substantiate the tribal or folk claims for its use as a safe hepatoprotective drug. Methods: The water, ethanol and n-hexane extracts of L. flexuosum were tested in vivo in Wistar rats. In acute toxicity model, rats received a single dose (5 g/kg) of water extract, ethanol extract, and n-hexane extract of L. flexuosum and kept under observation for 14 d. In subacute toxicity studies, animals were treated with daily doses (1 g/kg body weight) of water, ethanol and n-hexane extract respectively for 30 d. Results: In acute toxicity, the extracts administration up to a higher dose of 5 g/kg did not result in mortality or any change in behavior and biochemical parameters of the animals. Sub acute toxicity studies in rats showed that treatment with the extracts did not alter the serum biochemical and hematological parameters. Conclusions: The extracts were found to be devoid of any toxicity in acute (5 g/kg) and subacute (1 g/kg) toxicity evaluation in rats.

208-211

Authors: Vinod S Pawar, Shivakumar Hugar

Objective: To investigate the adaptogenic activity of methanolic extract of Trigonella foenum graecum seeds (METFGS) at 100, 250 and 500 mg/kg doses against anoxia stress tolerance in mice and immobilisation stress models. Methods: Withania somnifera (100 mg/kg, p.o.) was selected as reference standard and it showed significant effect in both stress models. The parameters like anoxia stress tolerance time was recorded in anoxia stress and estimation of biochemical marker levels and determination of organs weight were carried out in immobilisation stress model. Results: Concomitant treatment with METFGS at 500 mg/kg showed marked increase in anoxia stress tolerance time. But the lower doses (100 and 250 mg/kg) of METFGS produced mild increase in anoxia tolerance time. There was dose dependant significant reduction in biochemical parameters like serum glucose, cholesterol, triglycerides and BUN levels exhibited by test extract in immobilisation stress model as compared to stressed group. The stress induced increase in liver, adrenal gland weight and decrease in weight of spleen and testes were significantly reversed by the test extract at higher dose. But lower doses of extract caused the protective effect on weight of liver and adrenal gland only. Conclusion: The results from the present study indicate that METFGS possessed significant antistress activity

219-223

Authors: Lambole Vijay, Upendra Kumar

Objective: To investigate aqueous extract of bark of Moringa oleifera for normal wound healing and dexamethasone suppressed wound healing using incision, excision and dead space wound models in Wistar rats. Methods: In incision and dead space the extracts were applied daily topically till the 10th post wounding days while in excision model it was till the complete epithelialization process. Standard group were treated with Povidone iodine ointment topically daily. The breaking strength, percentage of wound contraction, period of epithelialization, dry granulation weight and hydroxyproline content were observed. Results: The aqueous extract of Moringa oleifera significantly increased the wound breaking strength in incision wound model. The aqueous extract treated wounds were found to epithelize faster and the rate of wound contraction was significantly increased as compared to control wounds. Significant increase in granulation breaking strength and hydroxyproline content in dead space wound was observed. The aqueous extract significantly decreased the antihealing activities of dexamethasone in all the wound models. Conclusions: The results indicated that the aqueous extract of Moringa oleifera promotes wound healing significantly and able to overcome the wound healing suppressing action of dexamethasone.

229-231

Authors: Pradeepa K, Krishna V, Venkatesh, Girish Kumar K, Santosh Kumar SR, Joy H Hoskeri, Gnanesh AU

Objective: To evaluate antinociceptive property of the leaf extract of Delonix elata (Linn.). Methods: Antinociceptive activity was evaluated by abdominal writhing and tail flick methods in Swiss albino mice. Acetyl salicylic acid was used as standard drug. Results: A dose dependent significant (P<0.01) antinociceptive activity was observed in three different concentrations 100, 200 and 300 mg/kg. Among them dose at 300 mg/kg, significantly (P<0.01) inhibited (74.94%) the nociception induced by acetic acid but less effective than the standard reference (84.06%). Tailflick test showed that the extract at the doses 300 mg/kg showed efficient results when compared to the other two doses (100 and 200 mg/kg) with increase in the latency to response of tail to the thermal stimulation. Conclusions: The significant antinociceptive activity of the leaf extract is due to the presence of a single active constituent in higher levels or due to the combined effect of more than one phytoconstituent. This investigation supported the ethnomedicinal claims of Delonix elata.

238-242

Authors: Pramod J Hurkadale, Pournima A Shelar, Siddhalingesh G Palled, Yuvaraj D Mandavkar, Ajay S Khedkar

Objective: To study the hepatoprotective activity of methanol and aqueous extracts of Amorphophallus paeoniifolius tubers against paracetamol induced liver damage in rats. Methods: Hepatotoxicity was induced by paracetamol and the biochemical parameters such as serum glutamic pyruvic transaminase (sGPT), serum glutamic oxaloacetic transaminase (sGOT) and serum alkaline phosphatase (sALP), serum bilirubin (SB) and histopathological changes in liver were studied along with silymarin and Liv-52 as standard hepatoprotective agents. Results: The phytochemical investigation of the extracts showed presence of carbohydrates, proteins, steroids and flavonoids. Pre-treatment of the rats with methanol and aqueous extract prior to paracetamol administration caused a significant reduction in the values of sGOT, sGPT, sALP and sB (P<0.01) almost comparable to the silymarin and Liv-52. The hepatoprotective was confirmed by histopathological examination of the liver tissue of control and treated animals. Conclusions: From the results it can be concluded that Amorphophallus paeoniifolius possesses hepatoprotective effect against paracetamol-induced liver damage in rats.

252-256

Authors: Sundaram Ravikumar, Samuel Jacob Inbaneson, Palavesam Suganthi

Objective: To identify the possible antiplasmodial compounds from Carica papaya (C. papaya), Pongamia glabra (P. glabra), Phyllanthus emblica (P. emblica) L, Phyllanthus acidus (P. acidus) L, Pisonia grandis (P. grandis) and Moringa pterygosperma (M. pterygosperma). Methods: The C. papaya, P. glabra, P. emblica L, P. acidus L, P. grandis and M. pterygosperma were collected from Ramanathapuram District, Tamil Nadu, India and the extraction was carried out in ethanol. The filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125 µg/mL) were tested for antiplasmodial activity against Plasmodium falciparum (P. falciparum). The potential extracts were also tested for their phytochemical constituents. Results: Of the selected plants species, the leaf extract of P. emblica L showed excellent antiplasmodial activity (IC50 35.09 µg/mL) followed by leaf extract of M. pterygosperma (IC50 42.36 µg/mL), leaf extract of C. papaya (IC50 46.23 µg/mL) and leaf extract of P. glabra (IC50 48.17 µg/mL). The flower extract of M. pterygosperma showed IC50 values of more than 100 µg/mL. Statistical analysis reveals that, significant antiplasmodial activity (P<0.01) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out and it shows that, there were no morphological changes in erythrocytes by the ethanolic extract of all the tested plant extracts. The in vitro antiplasmodial activity might be due to the presence of alkaloids, glycosides, carbohydrates, flavonoids, phenols, saponins, triterpenoids, steroids and tannins in the ethanolic extracts of tested plants. Conclusions: The ethanolic stem extracts of P. emblica L possess lead compounds for the development of antiplasmodial drugs.

260-264

Authors: Utpal Adhikari, Someshwar Singha, Goutam Chandra

Objective: To analyze the mosquito repellency and larvicidal activity of mature leaves of Swietenia mahagoni (S. mahagoni) against Culex quinquefasciatus (Cx. quinquefasciatus) Say in laboratory bioassay.Methods: Various concentrations of crude and petroleum ether extract of S. mahagoni mature leaves were exposed against all instars of Cx. quinquefasciatus larvae and the respective LC50, LC90, Regression equations and R values were calculated. Mosquito repellency was tested with chloroform: methanol (1:1, v/v) extract of mature leaves. Results: Cent percent mortality of 2nd instar mosquito larva was observed at 50 ppm concentration of petroleum ether extract after 72 h of exposure. 1st, 3rd and 4th instar larvae showed 96.66%, 90.00% and 60.00% mortality at 50 ppm concentration of petroleum ether respectively after 72 h of exposure. Preliminary qualitative phytochemical assay revealed the presence of saponins, alkaloids, tannin, flavonoids, and free glycoside bound anthroquinones. Chloroform: methanol extract showed repellency up to 2 h 15 min after application. No mortality was found in non target organisms, such as Gambusia affinis, Tadpole of Bufo and Chironomus larvae. Chloroform: methanol of mature leaves extract of S. mahagoni exhibits 100% repellency upto 2 h 15 min as no mosquito bites up to that time periods in the treated hands. Conclusions: Different solvent extracts of mature leaves of S. mahagoni can be effectively used as a potent ecofriendly biocontrol agent against larval form of Cx. quinquefasciat.

270-275

Authors: Bhavik kumar H Satani, Shri Hari Mishra

Objective: To generate pharmacognostic parameters of leaves of Heterophragma quadriloculare K. Schum since data on pharmacognostic and phytochemical characterization which provide means to authentic raw material or drug for medicinally important formulation are not available. Methods: The present study reports detailed pharmacognostic characters as identification parameters of the leaves which were subjected to morphological, histological, quantitative microscopic and chromatographic studies. Morphological, histological, quantitative microscopic were done using reported methods and chromatographic studies were performed using HPTLC. Results: Diagnostic features for identification of shape of leaves being compound, odd-pinnate, dorsiventral asymmetric at base with unicellular non-covering and glandular trichomes having unicellular stalk and bicellular head, actinocytic stomata and rhombus calcium oxalate crystals. Water soluble and alcohol soluble extractives were found to be (23.6±0.346)% w/w and (10.8±0.115)% w/w, respectively.Conclusions: The determinations from above studies establish the macro- and microscopic parameters which can be utilized for quick identification of the drug both as whole and in the powdered leaf materials and may helpful in laying down pharmacopoeial standards of this important member of traditional medicine.

284-289

Authors: S. Vijaya Kumar, Dhirendra B. Sanghai, C. Mallikarjuna Rao, C.S. Shreedhara

Objective: To standardize Melia azedarach L. by detailed study of macorscopical, microscopical and physiochemical parameters. Methods: Bark sample and dried powder of Melia azedarach were studied macroscopically and microscopically. Preliminary phytochemical investigation, physiochemical parameters of the bark sample as per WHO recommended guidelines for standardizations were also performed. Results: The detail microscopy revealed the presence of collapsed phloem, non-collapsed phloem, sieve elements, sieve plates, sieve tubes, companion cells, crystals and starch grains, narrow and wide fibres. Physiochemical parameters such as percentage of foreign matters, ash values, loss on drying, swelling index extractive values were determined. Total phenolic and flavonoid content were also determined. Preliminary phytochemical screening showed the presence of carbohydrates, tannins, terpenoids, glycosides, flavonoids and phenolic compounds. Conclusions: The macroscopic, microscopic and physiochemical analysis is useful in standardization of the Melia azedarach bark.

296-304

Authors: Sandhya S , Chandra Sekhar J, Vinod.K.R, Rao KNV, David Banji

Objective: The present research is envisaged on the preclinical studies of a polyherbal phytocomplex hair growth promoting cream incorporated with aqueous extracts of Trichosanthes cucumerina (T. cucumerina) Linn and Abrus precatorius (A. precatoius) Linn on enhancing the hair growth. Methods: Leaves of T. cucumerina and A. precatoius were separately extracted with distilled water and then subjected to preliminary chemical test. It was then made into a phyto-complex and finally formulated into a hair cream. The various evaluation parameters like organoleptic characters, presence of foreign particles/grittiness, determination of pH, irritancy, partition coefficient, drug content, diffusion studies, stability studies, sterility test, SEM, FTIR, solubility and hair growth promoting activity were performed. Results: Microscopic view of phyto-complex showed micelle structure. The formulated phyto-complex hair growth promoting cream had a smooth consistency with slightly acidic pH. FT-IR studies showed that the phytoconstituents were compatible with the excipients used. SEM studies revealed binding nature of the complex. The cream was found to be stable as the globule size, and was not altered on storage. Sterility testing proved the formulation to be free from microbial attack. The preclinical studies showed that the formulated 2% polyherbal phyto-complex hair growth promoting cream was an effective hair growth promoter as the results were analogous to that of minoxidil 2%. It was observed that percentage of hair follicles in the anagen phase increased considerably which predicts that the formulation can be used in alopecia. Conclusions: As the 2% polyherbal phytocomplex hair cream did not show any kind of irritancy or toxicity to the test animals it can be suggested that the prepared cream can be a better alternative for synthetic hair growth promoters.

308-311

Authors: Karthikeyan M, Umarul Mukhthar AK, Megha M, Shadeer Hamza P

Objective: To develop fast dissolving tablets of diclofenac in order to get fast relief from pain. Methods: Fast dissolving tablets of diclofenac potassium were prepared by direct compression method using Indion 214, Indion 234, Indion 244 and croscarmellose as superdisintegrants. Microcrystalline cellulose was used as diluents and mannitol, as sweetening agent. Tablets were evaluated for weight variation; weight variation of all the formulations was observed which were within the acceptable limit for uncoated tablets as per United States Pharmacopoeia. Results: Mechanical strength, the hardness of tablets was determined and was found to be in the range of 6.4 to 7.13 Kg/cm2 . In vitro disintegration time, the formulation containing indion214, indion234 andindion244 showed 34,31and 39 seconds value for in vitro disintegration respectively. Wetting time for all the formulations was found to be (4.425±0.25) to (5.75±2.25) seconds and the cumulative percentage drug release of formulations FD1- FD5. It was observed that in first 10 minutes, only 12.89% of drug was release from formulation FD5 without using superdisintegrants and while it was 88.56% in the case of formulation FD2. At the end of 24 minutes only 85.42 % of drug was released from the control tablet formulation FD5 whereas the cumulative release from other formulations FD1 and FD4 at the end of 12 minutes was 89.52 %and 79.29%, respectively. The results of in vitro disintegration time show that formulation containing Indion 244, showed the disintegration time of 29.66 seconds. Conclusions: Indion 244 was found to have super disintegrant property the addition technique is a best method for preparing fast dissolving tablets for rapid pain mangment.

316-322

Authors: S. Prabhakaran, R. Rajaram, V. Balasubramanian, K. Mathivanan

Objective: To screen the antifouling potential of various extracts from seaweeds (Ulva reticulate, Sargassum wightii, Halimeda macroloba), sea grasseses (Halodule pinifolia, Cymodocea serullata) and mangrove plants (Rhizophora apiculata, Rhizophora mucronata and Avicennia marina) against some marine fouling bacteria. Methods: The different species of seaweeds, seagrasses and mangrove samples were collected, washed, air dried and fine powdered samples were subjected to solvent extraction by cold steep method. The extracts fraction was eluted with the ethanol and subjected to FTIR. The biofilm forming bacteria were scrapped from the marine environment by biofilm formed PVC sheet. Among these ten strains isolated, four isolates (Flavobacterium sp., Bacillus sp., Cytophaga sp., Pseudomonas sp.) were chosen for this study. Results: Among the tested extracts, Avicennia marina limited the growth of Flavobacterium sp. (16 mm) and Bacillus sp. (20 mm) and the extracts of Rhizophora mucronata limited the growth of Flavobacterium sp. (18 mm) and Bacillus sp. (18 mm). While comparing the inhibition activity of all the extracts, mangrove plants extracts had higher inhibiting activity against primary biofilm forming bacteria than seaweeds and seagrasses. The inhibition activity was mainly correlated with the major functional groups [hydroxyl, amino, carbonyl and phosphoryl functionalities, aliphatic (fatty acids), NH2 (amide I & II)] of the extracts. Conclusions: The bioactive fractions from the above results indicates the occurrence of active constituents in the extracts of seaweeds, seagrasses and mangrove. It shows the improved antifouling activity against marine micro-fouling bacteria. These extracts can be used as the possible natural sources for anti-foulant.

329-333

Authors: Govindasamy C, M. Arulpriya, P. Ruban

Objective: To explore the antimicrobial compounds from the rhodophyceae family member algae Gracilaria corticata using 1 H and 13 C NMR spectral analysis. Methods: Antimicrobial activity was done by using the method of Prabhakar et al. Methanol and acetone extracts were tested against the human pathogens. The acetone extract was subjected to column chromatography using acetone as a solvent for purification and also the chemical analysis combined with 1 H and 13 C NMR analysis. Results: Among the two extracts, acetone extract showed higher activity than methanol extract. 1 H and 13 C NMR analysis showed the fraction extracted with acetone had polysaccharide like compounds. Conclusions: Altogether our observations suggest a triggering of antimicrobial activity by this polysaccharide like compound. Further the investigations are now in progress to purify and identify the structure of the compounds.

342-346

Authors: Karunamoorthy Manivannan, Perumal Anantharaman, Thangavel Balasubramanian

Objective: To evaluate the in vitro antioxidant capacity of a Chlorella marina (Butcher, 1952). Methods: Samples were tested for the total phenolic content, antioxidant activity, deoxy ribose radical scavenging activity and reducing power. Results: The methanolic extract was found to have high levels of phenolic (0.647±0.052 mg GAE/g), when compared to diethyl ether and hexane. The diethyl ether extract exhibited higher antioxidant potential as (0.816±0.366 mg AscAE/ g), higher percentage of deoxy ribose radical scavenging activity (0.399±0.004) and reducing power (2.814±0.014). Conclusions: The phenolic compounds were not a major contributor to the antioxidant capacities of these microalgae. This was very different from many other plant species like fruits, vegetables and medicinal plants. The microalgae could contain different antioxidant compounds from other plants.

352-357

Authors: J. Margaret Beula, M. Gnanadesigan, P. Banerjee Rajkumar, S. Ravikumar, M. Anand

Objective: To identify the antiviral, antioxidant and toxicological evalution of the mangrove plant from South East coast of India. Methods: In vitro antiviral analysis such as RT inhibition assay, HBsAg binding assay, HBV DNA polymerase Inhibitory assays were carried out with the leaf extract of Avicennia marina, Avicennia officinalis, Rhizophora mucronata and Rhizophora apiculata mangrove plants. Further, the in vitro antioxidant assays and in vivo toxicological assay was also carried out with the most potent Avicennia marina leaf extract. Results: Of the selected 4 mangrove plants, Avicennia marina leaf extract showed the minimum IC50 values with 403.91, 489.39 and 372.09 µg/mL concentrations against reverse transcriptase inhibition assay, HBsAg binding assay, HBV DNA polymerase Inhibitory assays, respectively. The antioxidant IC50 values were identified as 12.80±0.93, 640.06±34.93, 19.91±3.93 and 142.06±17.93 µg/mL concentrations against SOD, LPO, NO and DPPH assays, respectively. The phytochemical analysis of Avicennia marinaleaf extract showed the presence of reducing sugars, polyophenols, flavonoids and tannin. The LD50 value was 2 500 mg/kg bodyweight and the sub acute toxicity analysis did not show any significant changes in serum and haematological parameters when compared with the control animals. Conclusions: It can be concluded from the present findings that, the leaf extract of Avicennia marina can be further used as potential antiviral drug after completing the clinical trials.

368-372

Authors: RVignesh, M Srinivasan

Objective: To analyze the proximate composition, fatty acids, amino acids and nutritional composition of flour made from Tilapia (Oreochromis mossambicus) head and bones. Methods: Tilapia fish head and bone are the parts with unknown composition. Fish bones and heads were collected, cleaned and dried. The flours were prepared using 14-mesh stainless steel sieve and analysed further. Results: The results in 100 g of Tilapia head flour (THF) were composed of moisture [(5.89±0.01) %] protein [(32.59±0.02) %] total lipids [(0.2014±0.0002)%], and ash [(11.14±0.02) %]. The results in 100 g of Tilapia bone flour (TBF) were: moisture [(4.22±0.02)%], protein [(31.52± 0.02) %], total lipids [(0.8761±0.0002) %], and ash [(0.89±0.01)%]. The fatty acid composition of THF occurring in the highest proportions were alpha linolenic acid [(C18: 3; (2.4390±0.0025)%], stearic acid [(C18:0; (1.9883±0.0025)%], linoleic acid [(C18:2; (1.9860±0.0025)%], palmitic acid [C16:0; (1.1787 ±0.0002) %], oleic acid [(C18:1; (1.5468±0.0002)%] margaric acid acid [(C17:0 (0.4146±0.0002)%] and moroctic acid [(C18:4; (0.1141±0.0002)%]. The fatty acid compositions of TBF occurring in the highest proportions were stearic acid [(C18: 0; (0.7860±0.0025)%], oleic acid [(C18:1; (0.1141±0.0002)%], alpha linolenic acid [(C18: 3; (0.1141±0.0002)%], palmitic acid [(C16: 0; (0.1143±0.0025)%], linoleic acid [(C18:2; (0.0816±0.0002)%], margaric acid acid [(C18:0; (0.0088±0.0003)%] and moroctic acid [(C18:4; (0.0034±0.0003)%]. The major amino acids such as glutamic acid (0.9967%), aspartic acid (1.837%), lysine (1.048%) and leucine (0.807%) were found in THF and in TBF the major amino acids were found as glutamic acid (0.7865%), aspartic acid (0.9686%), lysine (1.342%) and leucine (0.7756%). The THF flour possessed seven essential minerals in milli grams per gram (mg/gm) such as calcium (56.7±0.2 mg), iron (3.098±0.002 mg), potassium (16.78±0.02 mg), magnesium (15.67±0.03 mg), copper (1.414±0.002 mg), sodium (34.67±0.03 mg) and zinc (0.343±0.003 mg). The TBF flour possessed calcium (67.76±0.02 mg), iron (2.41±0.02 mg), potassium (15.67±0.02 mg), magnesium (45.76±0.02mg), copper (0.131±0.001 mg), sodium (29.11±0.01 mg), and zinc (0.2415±0.0002 mg). Conclusions: The results indicate that the flours made from the fish head and bone of tilapia may result as a valuable alternative food in the human diet. This is the first study which reports the nutritive value of tilapia (Oreochromis mossambicus) byproducts such as heads and bones.

377-379

Authors: Vasanthi Rajendran, Sameer Hassan, Jerrine Joseph, Nagamiah Selvakumar, Vanaja Kumar

Objective: To do a comparative homology search for the 32 mycobacteriophages. Methods: This can be estimated as 3 400 proteins from 32 mycobacteriophages assuming each phage has 80-100 genes. The algorithm most widely used for homology detection in comparative genomics is Basic Local Alignment Search Tool (BLAST). Usually a stringent score cutoff is applied to distinguish putative homolog’s from possible false positive hits. As a consequence, some BLAST hits are discarded or put into insignificant hits that are in fact homologous. Assigning function to protein sequences is important. Here, we review the status of sequence (BLAST) based and domain based approaches to proteins that can provide functional insights such as database on Protein Families (PFAM). Results: The findings showed that 31% of proteins showed similarities with homologous protein with known function. Conclusions: In the present study only 31% of mycobacteriophage proteins functions were able to be predicted. Only for 6 proteins, the template structures were available but then they were not directly involved in phage lifecycle. Hence this emphasizes the importance of exploring the structures for mycobacteriophage proteins in order to understand their function and evolutionary significance. Ab initio methods for protein structure prediction can be only alternative for the rest of the proteins but accuracy of prediction is not highly dependable.

383-385

Authors: Jerrine Joseph, Sameer Hassan, Vasanthi Rajendran,Vanaja Kumar

Objective: To functionally annotate the protein sequences for 64 mycobacteriophage genomes using the ACLAME database and to discuss how structured information on mycobacteriophage encoded proteins helps global in silico analysis. Methods: Each of the protein sequences of 64 mycobacteriophage genomes were searched against ACLAME database Version 0.4 using BLAST and were further classified based on function. The default setting of ACLAME 0.4 BLAST was used. Results: Based on ACLAME, assignment of 72 protein functional families was done for the 64 sequenced genomes, and families were compared among and across genomes. Conclusions: The finding of the present study provides scope for designing novel approaches to decipher this huge chunk of unknown proteins by experimental methods.

392-400

Authors: C. Balachandran, V. Duraipandiyan, S. Ignacimuthu

Objective: To isolate active actinomycetes from soil samples of Northern Himalayas and study their culture characterization, protease production and cytotoxic effects on cancer cell line (A549). Methods: Forty six strains of actinomycetes were isolated from the soil collected from Northern Himalayas, India. Isolation of actinomycetes was performed by serial dilution plate technique. Forty six isolated actinomycetes cultures were grown in ISP 2 medium to study the morphology and biochemical characteristics. Isolated strains were studied for protease enzyme production in skim milk agar medium with solubilising capacity. Seven isolates were studied for melanin pigmentation and different NaCl concentration. Effects of environmental conditions on protease enzyme production of seven isolated strains were also studied at different pH, temperature and metal ions (毬-mercaptoethanol, dithiothreitol, iodoacetamide, MgSO4, CaCl2 and EDTA). Seven isolates were also studied for lytic enzyme activity using different bacteria and yeast such as Pseudomonas aeruginosa, Enterococcus feacalis, Escherishia coli, Candida albicans, Bacillus subtilis, Klebsiella pneumonia and Staphylococcus aureus. Results: Isolates ERIA-31 and ERIA-33 produced more protease enzyme activity in modified nutrient agar media compared to other actinomycetes cultures. ERIA-31 and ERIA-33 were tested for cytotoxic effect in human adenocarcinoma cancer cell line (A549). IC50 for ERIA-31 was 57.04 µg/mL and IC50 for ERIA- 33 was 55.07 µg/mL. Conclusions: Actinomycete being a protease producing bacteria has the potential for use in industrial purpose, pharmaceuticals, cytotoxic agent and its proteolytic activity. Isolates of ERIA-31 and ERIA-33 produced significant amount of protease enzymes.

406-410

Authors: Narayanapillai Udhayashree, Duraisamy Senbagam, Balakrishnan Senthilkumar

Objective: To isolate bacteriocin- producing lactic acid bacteria (LAB) and to asses its preservative effects on food products. Methods: Lactobacillus strain was isolated from chicken intestine using MRS broth (Hi-Media, India) at 37 °C for 48 h. The isolate was identified by 16s rRNA sequencing and phylogenetic analysis. Bacteriocin was produced, purified by ammonium sulphate precipitation and dialyzed. Antagonistic characteristics of bacteriocin were studied by agar well diffusion method. Bacteriocin stability was studied in different temperature, pH and enzymes. Biopreservative efficacy was determined for fish and fresh juice. Results: Lactobacillus fermentum(L. fermentum) strain isolated from chicken intestine was capable of producing bacteriocins, inhibiting the widest range of pathogenic bacteria. The bacteriocin was stable at 37 °C and acidic pH and proteinaceous in nature. Their biopreservative effect on refrigerated food products (apple juice and fish) show low count of colonies. Lactobacillus isolate was sequenced which shows high similarity with reference strain L. fermentum CECT 5716. Conclusions: Bacteriocin producing L. fermentum UN01 could be successfully used as biopreservative and to increase the shelf life of the food products.

418-422

Authors: Amrin Ali, Ritika Raj Shrivastava, Nirupama Agrawa

Objective: To observe their specificity, speciation and ecological equivalence by using previous and present records, from freshwater fish from Tanakpur to Sultanpur areas, during 2009 & 2010. Methods: Data is gathered from the published records to compare it with the present records and entered into a computer database. The taxonomy of monogenoideans follows and those of fish follows. The worms were gently scrapped from the gills of fish and counted manually. They were studied mostly live, under a phase contrast microscope (Olympus CX 41 U-DA 4E 03365 Japan) or fixed in 3% formalin, after being relaxed in lukewarm water. Results: These monogenoids have perfect ecological equivalence, as their macro habitat is geologically connected. They exhibit specificity at different taxonomic levels of the parasites and the hosts. Sympatric speciation of parasites and their host is evident in the present study. Conclusions: The monogenoidean fauna, in this region, has its species integrity in the past sixty to sixty five years. From this important body of study, it seems likely that host specificity among monogenoids from the fresh water fish is more widely spread than previously thought. Seasonal variations, probable controlling factor of infection, determine the intensity.

429-434

Authors: S Shanmugam, K Rajendran, K Suresh

Objective: To elucidate the medicinally important herbs and their role in the health cares of the villagers living in Sivagangai district of Tamil Nadu, India. Methods: Interviews and detailed personal discussions were conducted with the herbalist and local people to identify plants and collect the medicinal informations for 12 months (from June 2009 to May 2010). The medicinal important plants were botanically identified. Results: Totally 71 species of 61 genera belonging to 36 families were reported with ethnomedicinal values. Leaves are the mostly used part to prepare medicine. Generally fresh parts are used. Conclusions: Attention should be made on proper exploitation and utilization of these medicinally important plants.

439-445

Authors: Rajendra Yonzone, R. B. Bhujel, Samuel Rai

Objective: To find out the genetic resources, current ecological status and altitude wise distribution of medicinal plants diversity of Darjeeling Himalaya of West Bengal, India. Methods: The intensive field survey was conducted in the entire regions and necessary information were collected from the local inhabitants and literatures. Results: The present study revealed 218 medicinal plants species from 97 families with 195 genera distributed throughout the different altitudinal ranges of entire Darjeeling Himalaya. Of these, 6 plant species were from 3 500-45 00 m altitude, 22 from 1 800-3 500 m, 69 from 1 000-1 800 m and 121 from 130-1 000 m in the study area. There were 24 cultivated medicinal plants and the rest were wild. Conclusions: It is unique for rich, wide genetic diversity resources and distribution of medicinal plants in distinct climatic zones. However, the status of these some plant species are vulnerable, threatened, rare and critically endangered.

449-455

Authors: D. K. Patel, K. Patel, M. Gadewar, V. Tahilyani

Flavonoids are low molecular weight plant phytoconstituents, widely distributed in the leaves, seeds, bark and flowers of plants. It has various pharmacological activities and protects plants against ultraviolet radiation, pathogens, and herbivores. Most of the beneficial health effects of flavonoids are mainly due to its antioxidant properties. Galangin is a flavonol found in honey, Alpinia officinarum, Helichrysum aureonitens and in propolis. It showed anti-mutagenic, anticlastogenic, anti-oxidative, radical scavenging, metabolic enzyme modulating activity and effective against certain types of cancers. The present review described the pharmacological activity and bioanalytical aspects of galangin, which may be benificial to the researchers who wants to explore the hidden potential of galangin.

462-463

Authors: PV Chaitanya Varma, Shrikiran A Hebbar, Ramesh Y Bhat, Suneel C Mundkur, Karthick Annamalai, Sonia Bhatt

Antibodies against the Golgi complex (AGAs) have been reported rarely and are associated with disorders like systemic lupus erythematosus, Sjögren's syndrome and rheumatoid arthritis. We report a case of autoimmune haemolytic anemia with Antigolgi antibodies, the first such case in medical literature.

421-427

Authors: Santram Lodhi, Singhai AK

Objective: To evaluate the wound healing potential of fractions from ethanol extract of Martynia annua (M. annua) Linn leaves. Methods: Ethanol extract of M. annua Linn leaves was fractionate into three different fractions (MAF-A, MAF-B and MAF-C) which were screened for wound healing potential using two models: excision and incision on rats. The thin layer chromatography (TLC) profile of all fractions were analyzed and TLC of luteolin was also done. The PovidoneIodine Ointment was used as reference for comparision. Excision and incision wounds were created on dorsal portion of rats for study. Wound contraction, biochemical parameters (protein level and hydroxyproline level) and histopathological study were performed in excision wound model whereas incision model was used for determination of tensile strength. Results: The wound contraction and tensile strength of skin tissues were observed significantly greater in MAF-C fraction treated group than other two fractions (P<0.01). In excision wound method (on day 18) protein content and hydroxyproline were found significantly higher in MAF-C group than control group (P<0.01). Histopathological study also showed better angiogenesis, matured collagen fibres and fibroblast cells as compared with the control group. Conclusions: In conclusion, our findings suggest that fraction MAF-C from ethanol extract of M. annua leaves is found most effective in wound healing.

434-438

Authors: Endang Purwaningsih

Objective: To observe morphology of Auchenacantha galeopteri (A. galeopteri), Auchenacantha spinosa (A. spinosa) and Auchenacantha parva (A. parva) from Sukabumi and Ujung Kulon, Indonesia using scanning electron microscopy (SEM). Methods: Specimens for light microscopy examination were fixed with warm 70% alcohol, cleared and mounted in lactophenol for wet mounting. Drawings were made with the aid of a drawing tube attached to a Nikon compound microscope. Specimens for SEM examination were processed according to Bozzola. Measurements were given in micrometers (µm) as the average of findings, followed by the range in parentheses, unless otherwise stated. Results: The measurements of Auchenacantha spp are same with those of previously described. The striations of male and female A. galeopteri are widen towards outside and wavy. The mouth of female with 6 lips, each of them is wider at base than distal end in A. galeopteri and rectangular in A. spinosa. Both species have dentiform protrusion from inner surface of lips, such structure absent in A. parva, but the lips with transverse festoon like pattern at anterior end of each lip. Conclusions: Using SEM, the lip and the striation pattern of three species of Auchenacantha can be clearly distinguished. Sukabumi and Ujung Kulon are new locality of A. galeopteri and A. spinosa , and A. parva is the new record in Indonesia.

443-447

Authors: Mahendra Jain, Rakhee Kapadia, Ravirajsinh N Jadeja, Menaka C Thounaojam, Ranjitsinh V Devkar, SH Mishra

Objective: To evaluate the cytotoxicity and hepatoprotective potentials of extracts, fractions or isolated compound from the leaves of Feronia limonia (F. limonia). Methods: Qualitative phytochemical analysis of extracts, fractions or compound was performed by means of thin layer chromatography and spectroscopic assays. The % purity of compound was measured by analytical HPLC. Extracts, fractions or compound have been individually evaluated for their cytotoxicity effects (10, 20, 100, 250, 500, 750 and 1 000 µg/mL). Based on the inhibitory concentration (IC50) obtained from the cell viability assay, graded concentrations of extracts, fractions or isolated compound were assessed (10, 20, 50, 100, 200 µg/mL) for its hepatoprotective potential against CCl4-induced hepatotoxicity by monitoring activity levels of serum glutamatic pyruvatic transaminase (SGPT) and serum glutamic oxaloacetic transaminase (SGOT). Results: Results indicated that the methanol extract of F. limonia was non-toxic and hepatoprotective in nature as compared with the petroleum ether extract. The acetone fraction of methanolic extract also showed similar properties but the subsequent two fractions were cytotoxic. However, the pure compound isolated from the penultimate fraction of methanolic extract was non-toxic and hepatoprotective in nature. Biochemical investigations (SGOT, SGPT) further corroborated these cytological observations. Conclusions: It can be concluded from this study that F. limonia methanol extract, some fractions and pure isolated compound herein exhibit hepatoprotective activity. However, cytotoxicity recorded in the penultimate fraction and investigation of structural details of pure compound warrants further study.

452-455

Authors: Abdul Viqar Khan, Qamar Uddin Ahmed, M Ramzan Mir, Indu Shukla, Athar Ali Khan

Objective: To investigate the antibacterial potential of the polar and non-polar extracts of the seeds of Melia azedarach (M. azedarach) L. (Meliaceae) against eighteen hospital isolated human pathogenic bacterial strains. Methods: Petrol, benzene, ethyl acetate, methanol, and aqueous extracts at five different concentrations (1, 2, 5, 10 and 15 mg/mL) were evaluated. Disk diffusion method was followed to evaluate the antibacterial efficacy. Results: All extracts of the seeds demonstrated significant antibacterial activity against tested pathogens. Among all extracts, ethyl acetate extract revealed the highest inhibition comparatively. The present study also favored the traditional uses reported earlier. Conclusions: Results of this study strongly confirm that the seed extracts of M. azedarach could be effective antibiotics, both in controlling gram-positive and gram-negative human pathogenic infections.

461-467

Authors: Abdur Rashid, Mohammad Farooq Swati, Hassan Sher, Mohammad N Al- Yemeni

Objective: To determine the present status of plant communities and their possible association with the habitat in Malam Jabba, Swat, Pakistan. Methods: A study on the phytoecology was conducted in various ecologically important sites of Malam Jabba, Swat, Pakistan from 2002 to 2004. The altitude of these sites ranged from 1 200 m to 3 200 m. Quadrat method was used for evaluation of plants communities and the data on these attributes was converted to relative values. The plant communities were named after 3 leading species with highest importance values. Biological spectrum of the flora based on the life form was prepared by following Raunkiar’s life form classes. Results: The floristic composition and structure of the study area were found to be 200 species belonging to 75 families. Asteraceae, Lamiaceae and Poaceae were important families in the study area. The biological spectrum showed that therophytic and hemicrytophytic life form and micro-nonophyllous leaf sizes were dominant in the area. The air and soil temperatures were decreasing with increasing elevation. Both the air and soil temperatures were relatively higher in south slopes than on the northeast slopes. The vegetation analysis of the area indicated eleven plant communities around the area. The present vegetation is the relics of moist temperate coniferous forest in the area. The communities reflect highly deteriorated conditions. Both the structure and composition of the surrounding vegetation were associated with the types of habitats. Conclusions: The conservation of the remaining populations of the reported communities will be best achieved by proper time of sustainable harvesting. It is only possible with the participation of local communities.

472-477

Authors: H Nur Aida, Hamady Dieng, Abu Hassan Ahmad, Tomomitsu Satho, AT Nurita, MR Che Salmah, Fumio Miake, B Norasmah

Objective: To generate life table characteristics for the dengue vector Aedes albopictus (A. albopictus) under uncontrolled conditions, incorporating both the aquatic and the adult stages. Methods: Ten females derived from wild pupae were allowed to fully blood-feed on restrained mice. 774 eggs were hatched in seasoned water. F1 larvae were followed for development until their F2 counterparts emerged as adults. Some population parameters were monitored (F1) or estimated (F2). Results: A. albopictus exhibited increased fecundity and egg hatch success. Immature development was quick. Immature survival was high, with lowest rate in the pupal stage. Adult emergence was about 81% and sex ratio was close to 1:1. Generational mortality (K) was about 28%. A high proportion of females completed a reproductive cycle and the obtained parity rate was predicted to lead to higher fecundity in the next generation. Conclusions: It can be concluded that natural A. albopictus populations in Penang seem largely determined by quick development in combination with low immature loss and increased oviposition.

482-487

Authors: Subhankari Prasad Chakraborty, Santanu KarMahapatra, Sabyasachi Das, Somenath Roy

Objective: To evaluate the free radical generation and antioxidant enzymes status in murine peritoneal macrophage during in vitro amikacin resistant Pseudomonas aeruginosa (ARPA) treatment with different time interval. Methods: Peritoneal macrophages were treated with 1× 108 CFU/mL ARPA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and super oxide anion generation, NO generation, reduced glutathione level and antioxidant enzymes status were analyzed. Results: Super oxide anion generation and NO generation got peak at 12 h, indicating maximal free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during ARPA transfection. Reduced glutathione level and antioxidant enzymes status were decreased significantly (P<0.05) with increasing time of ARPA transfection. All the changes in peritoneal macrophages after 12 h in vitro ARPA transfection had significant difference (P<0.05). Conclusions: From this study, it may be summarized that in vitro ARPA infection not only generates excess free radical but also affects the antioxidant system and glutathione cycle in murine peritoneal macrophage.

491-492

Authors: Youssefi Mohammad Reza, Rahimi Mohammad Taghi

Objective: To survey malaria prevalence in Sarbaz from April 2009 to October 2010. Methods: Epidemiological data of 1 464 confirmed malarial patients were analyzed according to demographic status, sex, age, nationality, isolated species and residence place. Results: The majority of patients were male 950 (64.8%) but 514 (35.2%) were female. 82.5% of patients were Iranian, 14% Pakistani immigrants, and 3.5% Afghan immigrants. Data collected showed that 90% of isolated species were Plasmodium vivax, 7.8% Plasmodium falciparum, and 2.2% Plasmodium malariae and mixed species. Conclusions: Therefore, it is crystal clear that refugees should be prohibited by government and controlled by experts in health centers in order to campaign effectively with this life threating disease.

496-501

Authors: Kamal SM, Rashid AKM M, Bakar MA, Ahad MA

Anthrax is a zoonotic disease caused by Bacillus anthracis. It is potentially fatal and highly contagious disease. Herbivores are the natural host. Human acquire the disease incidentally by contact with infected animal or animal products. In the 18th century an epidemic destroyed approximately half of the sheep in Europe. In 1900 human inhalational anthrax occured sporadically in the United States. In 1979 an outbreak of human anthrax occured in Sverdlovsk of Soviet Union. Anthrax continued to represent a world wide presence. The incidence of the disease has decreased in developed countries as a result of vaccination and improved industrial hygiene. Human anthrax clinically presents in three forms, i.e. cutaneous, gastrointestinal and inhalational. About 95% of human anthrax is cutaneous and 5% is inhalational. Gastrointestinal anthrax is very rare (less than 1%). Inhalational form is used as a biological warefare agent. Penicillin, ciprofloxacin (and other quinolones), doxicyclin, ampicillin, imipenem, clindamycin, clarithromycin, vancomycin, chloramphenicol, rifampicin are effective antimicrobials. Antimicrobial therapy for 60 days is recommended. Human anthrax vaccine is available. Administration of anti-protective antigen (PA) antibody in combination with ciprofloxacin produced 90%-100% survival. The combination of CPG-adjuvanted anthrax vaccine adsorbed (AVA) plus dalbavancin significantly improved survival.