Screening of antimicrobial potential of polysaccharide from cuttlebone and methanolic extract from...

8/13/2019 Screening of antimicrobial potential of polysaccharide from cuttlebone and methanolic extract from body tissue of

1/5

S244

Document heading

Screening of antimicrobial potential of polysaccharide from cuttlebone

and methanolic extract from body tissue of Sepia prashadiWinkworth,

1936

Pasiyappazham Ramasamy, Aruldhason Barwin Vino, Ramachandran Saravanan, Namasivayam Subhapradha, Vairamani

Shanmugam, Annaian Shanmugam*

Centre of Advanced Study in Marine Biology Faculty of Marine Sciences, Annamalai University Parangipettai - 608 502, India

Asian Pacific Journal of Tropical Biomedicine (2011)S244-S248

Asian Pacific Journal of Tropical Biomedicine

journal homepage:www.elsevier.com/locate/apjtb

*Corresponding author: Annaian Shanmugam, Centre of Advanced Study in MarineBiology Faculty of Marine Sciences, Annamalai University Parangipettai - 608502, India. Tel: +91-9443043597 E-mail: [email protected] Foundation Project: Supported by Centre for Marine Living Resources and Ecology

(CMLRE), Ministry of Earth Science (grant No. MoES/CMLRE/10A-4(6)/08).

1. Introduction

The cephalopod which includes the Nautilus, cuttlefishes,squids and octopods, is the most advanced class of phylum:Mollusca adapted to a swimming existence. They areexclusively marine, diverse in form, size and nature[1]andoccupy littoral and benthic to pelagic environments of allworld oceans. Cephalopods are considerably important as afood resource as well as in scientific investigations. Thereis an ever continuous and urgent need to discover newantimicrobial compounds with diverse chemical structures

and novel mechanisms of action due to the alarmingincrease that has been witnessed in the incidence of bothnew and reemerging infectious diseases. A further bigconcern is the development of resistance to the antibioticsin current clinical use[2].

Although antibiotics are life saving drugs but nowadays,as a result of careless and promiscuous use of antibiotics,various pathogenic microbes are gaining resistances.Among marine invertebrates, cephalopods belong to a

molluscan group comprising 700species in which bacterialassociations have been known for a long time and caninclude the reproductive organs (accessory nidamentalglands) of myopsids, sepiolids and sepiids and the lightorgan of sepiolids[3-5]. In recent years, human pathogenic microorganisms havedeveloped resistance in response to the indiscriminate useof commercial antimicrobial drugs commonly employedin the treatment of infectious diseases. This situation,the undesirable side effect of certain antibiotics, and theemergence of previously uncommon infections, has forced

scientists to look for new antimicrobial substitutionsfrom various sources such as from plant origin andanimal origin[6]. Marine invertebrates offer a source ofpotential antimicrobial drugs[7-9]. Discovered bioactivecompounds in molluscs were identified essentially aspeptide, depsipeptide, sterols, sesquiterpene, terpenes,polypropionate, nitrogenous compounds, macrolides,prostaglandins and fatty acid derivatives, sterols,miscellaneous compounds and alkaloids; they all presentedspecific types of activities[10-12]. Defer et al[13]reported theantibacterial and antiviral activities in three bivalve and twogastropod marine molluscs (Cerastoderma edule,Ruditapesphilippinarum, Ostrea edulis,andBuccinum undatum).

Studies of antimicrobial activity provide valuable

ARTICLE INFO ABSTRACT

Article history:Received 15August 2011Received in revised form 8September 2011Accepted 1 October 2011Available online 15 October 2011

Keywords:

Antimicrobial activityCephalopodPolysaccharideCuttlebone

Objective: To evaluate the antimicrobial activity of polysaccharide from cuttlebone andmethanolic extract from body tissue of Sepia prashadi, against ten human pathogenic bacteriaand five fungi. Methods:The activity of polysaccharide and methanolic extract was investigatedagainst Vibrio cholerae, Pseudomonas aeruginosa, Klebsiella pneumoniae, Vibrio alginolyticus,Staphylococcus aureus, Vibrio parahaemolyticus, Streptococcus sp., Streptococcus pneumoniae,Salmonella sp. and Escherichia coli, and five fungal strains such as Alternaria alternata,Candida tropicalis, Penicillium italicum, Fusarium equisetiand Candida albicanusing discdiffusion method and minimum inhibitory concentration (MIC)were also calculated. Results:Both polysaccharide and methanolic extract was active against gram positive than that of gramnegative pathogenic bacteria but inactive against fungi. The MICof both the extract rangingfrom 60to 100mg/mL. Conclusions:These results suggest that cephalopod polysaccharide andmethanolic extract possess relatively good antibacterial activity.

Contents lists available at ScienceDirect


2/5

Pasiyappazham Ramasamy et al./Asian Pacific Journal of Tropical Biomedicine (2011)S244-S248 S245

information for new antibiotic discoveries and give newinsights into extraction of bioactive compounds fromaquacultured molluscs. Romanenko et al[14] investigatedthe isolation, phylogenetic analysis and screening ofmarine mollusc-associated bacteria for antimicrobial,hemolytic and surface activities. In most of the publicationsconcerning antimicrobial activity in molluscs, either singlebody component alone, like haemolymph and egg masses, or

extracts of whole body tissues have been tested for activity.Typhoid fever is a more classical systemic infectioncaused by the typhoid bacillus,Salmonella entericaserovarTyphi, the most common cause of enteric fever, which alsoincludes paratyphoid fever caused by Salmonella paratyphi.With an estimated 16-33million cases resulting in 500000to 600 000 deaths annually in endemic areas, the WHOidentifies typhoid as a serious public health problem[15].Due to the permanent resistance of the microorganisms toavailable drugs, continuous search for new antimicrobialsis a scientific challenge. In our continuous search ofantimicrobial agents from natural sources, this studywas designed to assess the antimicrobial activity of thepolysaccharides extracted from cuttlebone and methanolicextract from body tissue of Sepia prashadi (S. prashadi).

2. Materials and methods

2.1. Sampling and identification

The specimen (S. prashadi)were collected from Cuddalorelanding centre (Lattitude 1042 N; Longitude 7946 E), southeast coast of India. The Publication of Roperet al, Jothinayagam and Shanmugam et alwere used inidentification[16-18].

2.2. Preparation of polysaccharide extract from cuttlebone

The polysaccharide extact was prepared using the methoddescribed by Okutani et al[19].The air dried cuttleboneswere pulverized and washed with acetone. The powderwas extracted with hot 10mMEDTAsolution and filteredusing Whatman No. 1 filter paper with hyflo super-cel.Then saturated barium hydroxide solution was added to thefiltrate. The precipitate obtained after standing overnightwas collected on Whatman No. 1paper with hyflo super-cel and washed with water. The precipitate was dissolved in10mMEDTAsolution and was dialyzed against deionizedwater. The dialysate was freeze-dried which was then used

in the present investigation.2.3. Preparation of methanolic extracts from the body tissues

The methanolic extracts of the body tissues were preparedby following the method of Elyet al[20].S. prashadiwerebrought to laboratory; body tissues were removed, cutinto small pieces and homogenized (REMI, RQ-127A)andextracted with MeOHat room temperature fore 24-48h.Then the methanolic extract was centrifuged to collect thesupernatant and concentrated under vacuum in a rotaryevaporator at low temperature.

2.4. Microbial cultures

Ten species of bacteria and five species of fungi were usedas test organisms. The bacterial strains contains Gram-positive strains: Streptococcus sp., Streptococcus pneumoniae(S. pneumoniae), Staphylococcus aureus (S. aureus); Gram-negative strains:Escherichia coli (E. coli), Vibrio cholerae(V. cholerae), Vibrio alginolyticus (V. alginolyticus), Vibrioparahaemolyticus (V. parahaemolyticus), Pseudomonas

aeruginosa (P. aeruginosa ), Klebsiella pneumoniae (K.pneumoniae)and Salmonella sp..The fungal strains containsAlternaria alternata, Candida tropicalis, Penici lliumitalicum, Fusarium equiseti and Candida albicans. Allthe bacterial and fungal strains were clinical isolates,obtained from the Raja Muthaiah Medical College Hospital,Annamalai University, Annamalai Nagar, India.

2.5. Inoculum preparation for bacteria

Nutrient broth was prepared and sterilized in an autoclaveat 15lbs pressure for 15min. All the ten bacterial strainswere individually inoculated in the sterilized nutrient brothand incubated at 37

for 24h. Mueller Hinton agar (MHA,

Himedia)was prepared, sterilized in an autoclave at 15 lbspressure for 15minutes and poured into sterile petridishesand incubated at 37 for 24h. The 24 hour-old bacterialbroth cultures were inoculated in the petridishes by using asterile cotton swab.

2.6. Inoculum preparation for fungi

Czapek dox (Hi-media)broth was prepared and sterilizedin an autoclave at 15 lbs pressure for 15min. Five fungalstrains were inoculated in the broth and incubated at 37for72h. The sterilized Czapek dox agar was poured into sterilepetridishes and incubated at 37 for 3days. The 72hour

-old fungal broth cultures were inoculated in the petridishesusing a sterile cotton swab.

2.7. Disc diffusion method

Antibacterial and antifungal activity was determinedfollowing the method of El-Masry[21]. The stock solution ofpolysaccharide and methanolic extracts was prepared at aconcentration of 100mg/mL.Sterile antimicrobial disc (Hi-media)was impregnated with 50Lof polysaccharide andmethanolic extract of the four concentrations tested. Positivecontrol disc containing 50Lof tetracycline (1mg/mL)andas negative control, 50Lof 10mMEDTAand methanol

were used. These impregnated discs were allowed to dry atlaminar air flow chamber for 3hours, and were placed atthe respective bacterial and fungal plates and incubatedat 37 for 24h for bacteria and 72hours for fungi. Thediameter (mm)of the growth inhibition halos produced bythe polysaccharide and methanolic extract of cephalopodswas examined. Result was calculated by measuring the zoneof inhibition in millimeters. All the tests were performed intriplicate.

2.8. Determination of the minimum inhibitory concentration

The polysaccharide and methanolic extracts were selectedfor the determination of MIC following the method of


3/5

Pasiyappazham Ramasamy et al./Asian Pacific Journal of Tropical Biomedicine (2011)S244-S248S246

Rajendran and Ramakrishnan[22].

2.9. Statistical analysis

Data on the inhibitory effects of polysaccharide andmethanolic extracts of cephalopods was analyzed by one-way analysis of variance (ANOVA)using SPSS-16versionsoftware followed by Duncuns multiple range test (DMRT)

and Mean SEM. Pvalues at


4/5

Pasiyappazham Ramasamy et al./Asian Pacific Journal of Tropical Biomedicine (2011)S244-S248 S247

4. Discussion

In recent years, great attention has been paid to thebioactivity of natural products because of their potentialpharmacological utilization. Most homeopathic medicinesare either of plant or animal origin. Several moleculesextracted from marine invertebrates, including bivalves,possess broad spectrum antimicrobial activities, affecting

the growth of bacteria, fungi and yeasts[23]. The overallobjective of the current study was to compare the ability ofantibacterial and antifungal activity of methanolic extractof the body tissue and polysaccharide extract from thecuttlebone of S. prashadi. The results of the present studyclearly showed that the 75% and 100 % concentrationsexhibited appreciable antibacterial activity against humanpathogens (Tables 1& 2). Antibacterial activity has previously been described in awide range of molluscan species such as oyster (Crassostreavirginica), mussel (Mytilus edulis and Geukensia demissa),muricid mollusc (Dicathais orbita)and sea hare (Dolabellaauricularia)[24-26]. Prem Anand and Patterson Edwardreported moderate antibacterial and antifungal activity from

the extracts of five species of Cypraea namely C. errones,C. arabica, C. onyx, C. tigris and C. vitellas[27] . Pattersonand Murugan reported the broad spectrum of antibacterialactivity for aqueous extract of ink of the cephalopods Loligoduvaucelii (L. duvaucelii) and Sepia pharaonisagainstnine human pathogens[28]. However the majority of marineorganisms are yet to be screened for discovering usefulantibiotics.

There are only a few studies carried out hitherto on theantibacterial activity of the internal bone of cephalopods.Barwin Vino [29], for the EDTAextract (polysaccharides)of Doryteuthis sibogae (D. sibogae )gladius reported 10mm inhibition zone against E. coliand K. pneumoniae, 9mm inhibition zone against S. aureusand 7mm againstSalmonella typhi (S. typhii)(excluding the disc size of 5mmdia). Whereas the EDTAextract of L. duvauceligladiusshowed only low activity i.e., 5mm against P. aeruginosa,4mm against S. typhiand E. coli (excluding the disc sizeof 5mm dia). At the same time, the gladius extract of boththe species showed no activity against V. cholerae. Thepolysaccharide extract from the gladius of D. sibogaerecorded potent antibacterial activity against all thebacterial strains mentioned above and at the same time thepolysaccharide of theL. duvauceliigladius extract recordedonly low activity. The polysaccharide extract from thegladius ofL. duvaucelishowed activity against the fungi suchas Aspergillus fumigatus(A. fumigatus), Aspergillus flavus(A. flavus)and Rhizopus sp.; whereas gladius extract of D.sibogaereported activity againstA. fumigatusandRhizopussp. only. But at the same time both the species showed noactivity againstCandida sp. at all the concentrations tested.

Shanmugam et al[30]observed that the antibacterial activitywas predominant among cuttlebone extracts (using EDTA)of the cuttlefishes such asSepia aculeata (S. aculeata)andSepia brevimana (S. brevimana)against almost all the 9pathogenic bacterial strains tested viz., Bacillus subtilis,

E. coli, K. pnemoniae, S. aureus, V. parahaemolyticus,V. cholerae, S. typhi, P. aeruginosa and Shigella sp. Theactivity was recorded in almost all the concentrations exceptin negative control. The cuttlebone extract of S. aculeataand S. brevimanaagainst four fungal stains such as A.fumigatus, A. flavus, Candida sp. andRhizopus sp. showedthe maximum antifungal activity in 100%concentration and

the activity was found to be in an increasing order from thelower to higher concentration. On comparison the activitywas higher in the cuttlebone extract of S. aculeata than S.brevimana. But in the present study at 100%concentration,the polysaccharide extract from the cuttlebone reported (131.53)mm of inhibition zone against V. parahaemolyticusandthe methanolic extract from the body tissue showed (121.25)mm of inhibition zone against S. aureus. This highest activityof polysaccharides may be due to the destruction of cellwall of V. parahaemolyticusand S. aureus. This result wasconsistent with that of oleoyl-chitosan, which could destroycell wall of S. aureusand lead to intracellular componentsbeing released[31]. Water-soluble intracellular protein of S.aureustreated with the polysaccharide increased rapidly,which indicated that cell wall and membrane of S. aureusmight be disrupted by the polysaccharide [32]. When compared to the above mentioned studies,polysaccharides (EDTA)extract from S. prashadicuttleboneexhibited better activity. In the present study the activitywas also dose dependent. At the same time, the zone ofinhibition was relatively higher than that of the previousabove study. However there exists a difference in theactivity shown by the compound(s)present in the extracts,in laboratory studies and natural environment while may bedue to their varying concentration present in the extractsused in both the places[33,34]. Although different species and experimental procedureswere used in the different studies, they indicated the highfrequency of detectable antimicrobial activity in marinemolluscs. These results enforce the idea that cephalopods area source to be considered in the discovery of new substancesfor drug development. In the present investigation, thereexists antibacterial against different bacterial strains. Butat the same time no antifungal activity was recorded in bothpolysaccharide and methanolic extracts studied. Premanandand Patterson also reported no antifungal activity in themethanolic extracts fromDidemnum psammathodes[27]. The

known antimicrobial mechanisms associated to each groupof chemical to which the isolated compounds belong mayexplain the antimicrobial potency of the crude extracts andcompounds from Vismia laurentii. Membrane disruptioncould be suggested as one of the likely mechanisms of actionof friedelin[35]. In the present study a wide spectral antibacterial activityhas been recorded in almost all the concentrations. Oncomparison the maximum zone of inhibition was obtained forthe methanolic extract than the polysaccharide extract fromS. prashadiwhich explains and supports the presence ofactive principle in both the methanolic and polysaccharideextracts obtained from the body tissue and cuttlebone of S.prashadi. Further the dose as well as the concentration of the

active principle in the extract shows either bactericidalorbacteriostaticaction against the bacteria i.e. a low dose of abactericidalantibacterial agent may only inhibit bacterialgrowth, while a high dose of a bacteriostaticantibacterialagent will be bactericidal[36]. Further investigation on thepurification and chemical elucidation of the active principlepresent in both the extracts shall pave the way for thedevelopment of either the base or a new drug itself in future.

Conflict of interest statement

We declare that we have no conflict of interest.


5/5

Pasiyappazham Ramasamy et al./Asian Pacific Journal of Tropical Biomedicine (2011)S244-S248S248

Acknowledgements

Authors are thankful to the Director & Dean, CASin MarineBiology, Faculty of Marine Sciences, Annamalai Universityfor providing with necessary facilities, and to the Centre forMarine Living Resources and Ecology (CMLRE), Ministry ofEarth Science, Cochin for providing the financial assistance.Grant No. of this project- MoES/CMLRE/10A-4(6)/08.

References

[1] Worms J. World fisheries for cephalopods. A synoptic overview.In: Caddy JF, editor.Advances in sssessment of world cephalopodresources.Virginia: Food and Agriculture Organization of theUnited Nations; 1983, p. 231-452.

[2] Ilhan S, Savarolu F, olak F. Antibacterial and antifungalactivity of Corchorus olitoriusL. (Molokhia)Extracts. Inter J Nat

Engi Sci2007; 1(3): 59-61.[3] Bloodgood RA. The squid accessory nidamental gland, ultra

structure and association with bacteria. Tissue Cell 1977; 9:197-208.

[4] Pichon D, Gaia V, Norman MD, Boucher-Rodoni R. Phylogenetic

diversity of epibiotic bacteria in the accessory nidamental glandsof squids (Cephalopoda: Loliginidae and Idiosepiidae).Mar Biol2005; 147(6): 1323-1332.

[5] Nishiguchi MK. Host-symbiont recognition in the environmentallytransmitted sepiolid squid-Vibrio mutualism.Microb Ecol2002;44:10-18.

[6] Ramachandran M, Thirumalai T, Vinothkumar P. Antibacterialactivity of various leaf extracts of Merremia emarginataEKElumalai.Asian Pac J Tropical Biomed 2011; 1: 406-408.

[7] Bansemir A, Blume M, Schrder S, Lindequist U. Screeningof cultivated seaweeds for antibacterial activity against fishpathogenic bacteria. Aquaculture2006; 252: 79-84.

[8] Mayer AMS, Rodriguez AD, Berlinck RGS, Hamann MT. Marinepharmacology in 2003-4: marine compounds with anthelminticantibacterial, anticoagulant, antifungal, anti-inflammatory,antimalarial, antiplatelet, antiprotozoal, antituberculosis, andantiviral activities; affecting the cardiovascular, immune andnervous systems, and other miscellaneous mechanisms of action.Comp Biochem Physiol C Toxicol Pharmacol2007; 145: 553-581.

[9] Jayaraj SS, Thiagarajan R, Arumugam M, Mullainadhan P.Isolation, purification and characterization of [beta]-1,3-glucanbinding protein from the plasma of marine mussel Perna viridis.Fish Shellfish Immunol 2008; 24: 715-725.

[10] Maktoob A, Ronald HT. Handbook of natural products frommarine invertebrates: Phylum Mollusca Part I.Newark: HarwoodAcademic Publishers; 1997, p. 1-288.

[11] Balczar JL, Blas Id, Ruiz-Zarzuela I, Cunningham D, Vendrell D,Mzquiz JL. The role of probiotics in aquaculture. Vet Microbiol2006; 114: 173-186.

[12] Blunt JW, Copp BR, Munro MHG, Northcote PT, Prinsep MR.Marine natural products.Nat Prod Rep 2006; 23: 26-78.

[13] Defer D, Bourgougnon N, Fleury Y. Screening for antibacterialand antiviral activities in three bivalve and two gastropod marinemolluscs.Aquaculture2009; 293: 1-7.

[14] Romanenko LA, Uchino M, Kalinovskaya NI, Mikhailov VV.Isolation, phylogenetic analysis and screening of marine mollusc-associated bacteria for antimicrobial, hemolytic and surfaceactivities.Microbiol Res2008; 163: 633-644.

[15] World Health Organization. Diarrhoeal Diseases. Geneva: WHO;[Online] Available at: www.who.int/vaccine research/diseases/diarrhoeal/en/index7/html (accessed on 16March 2009).

[16] Roper CFE, Sweeney MJ, Nauen CE. Cephalopods of the world.Rome, Italy:Food and Agriculture Organization;1984; 3: 1-277.

[17] Jothinanayagam JT. Cephalopods of the Madras coast, zoological

survey of India. Tech Monogra;1987; 15: 85.[18] Shanmugam A, Purushothaman A, Sambassivam S, Vijayalaksmi

S, Balasubramaniyan T. Cephalopods of Parangipettai coast, Eastcoast of India: Monograph. India, Annamalai University: Centre ofAdvanced Study in Marine Biology, 2002; p.45.

[19] Okutani K, Morikawa N. Gel filtration and sugar constituent of thepolysaccharide extracted from the internal shell of squids. BullJap Soc Fish1978; 44: 369-372.

[20] Ely R, Tilvi Supriya C, Naik G. Antimicrobial activity of marine

organisms collected off the coast of South East India.J Exp MarBiol Ecol 2004; 309: 121-127.

[21] El Masry HA, Fahmy HH, Abdelwahad ASH. Synthesis andantimicrobial activity of some new benzimidazole derivatives.Molecules2000; 5: 1429-1438.

[22] Rajendran NK , Ramakrishnan J. In vi tr o evaluation ofantimicrobial activity of crude extracts of medicinal plants againstmulti drug resistant pathogens. Biyoloji Bilimleri Aratrma

Dergisi2009; 2(2): 97-101.[23] Mitta G, Hubert F, Dyrynda EA, Boudry P, Roch P. Mytilin B

and MGD2, two antimicrobial peptides of marine mussels: genestructure and expression analysis.Dev Comp Immunol2000; 24:381-393.

[24] Anderson RS, Beaven AE. Antibacterial activities of oyster

(Crassostrea virginica)and mussel (Mytilus edulisand Geukensiademissa)plasma.Aquat Living Resour 2001; 14: 343-349.[25] Benkendorff K, Bremner JB, Davis AR. Indole derivatives from the

egg masses of Muricid molluscs.Molecules2001; 6: 70-78.[26] Prem Anand T, Rajaganapathi J, Patterson Edward JK .

Antibacterial activity of marine molluscs from Portonovo region.Indian J Mar Sci 1997; 26: 206-208.

[27] Prem Anand T, Patterson Edward JK. Antimicrobial activity inthe tissue extracts of five species of cowries Cypreasp. (Mollusca:Gastropoda)and an ascidianDidemnum psammathodes (Tunicata:Didemnidae).Indian J Mar Sci 2002; 25: 239-242.

[28] Patterson EJK, Murugan A. Screening of cephalopods forbioactivity.Phuket Mar Biol Cent Spec Pubi2000; 21: 1253-256.

[29] Barwin Vino A. Studies on cephalopods with reference topolysaccharides M. Phil. Dissertation. Porto Novo, India:Annamalai University; 2003, p. 70.

[30] Shanmugam A, Mahalakshmi TS, Barwin Vino A. Antimicrobialactivity of polysaccharides isolated from the cuttlebone ofSepia aculeata (Orbingy, 1848)and Sepia brevimana (Steenstrup,1875): An approach to selected antimicrobial activity for humanpathogenic microorganisms. J Fish Aquat Sci2008a; 3(5): 268-274.

[31] Xing K, Chen XG, Kong M, Liu CS, Cha DS, Park HJ. Effect ofoleoyl-chitosan nanoparticles as a novel antibacterial dispersionsystem on viability, membrane permeability and cell morphologyofEscherichia coli and Staphylococcus aureus. Carbohydr Polym2009; 76(1): 17-22.

[32] He F, Yang Y, Yang G, Yu L. Studies on antibacterial activityand antibacterial mechanism of a novel polysaccharide fromStreptomycesvirginia H03. Food Control2010; 21: 1257-1262.

[33] Kelman D, Kashman Y, Rosenberg E, Kushmaro AI, Loya Y.Antimicrobial activity of Red Sea corals. Mar Biol 2006; 149:357-363.

[34] Mirnejad R, Jeddi F, Kiani J, Khoobdel M. Etiology of spontaneousbacterial peritonitis and determination of their antibioticsusceptibility patterns in Iran. Asian Pac J Trop Dis2011; 1(2):116-118.

[35] Kuete V, Nguemeving JR, Beng VP, Azebaze AGB, Etoa FX,Meyer M, et al. Antimicrobial activity of the methanolic extractsand compounds from Vismia laurentiiDe Wild (Guttiferae). J

Ethnopharmacol2007; 109: 372-379.[36] Zarakolu P, Coplu N, Arslanturk A, Levent B, Guvener E.

Comparison of inhibitory and bactericidal activities ofcefoperazone +sulbactam against Psedomonas aeruginosa strains.

Tr J Med Sci1999; 29: 607-609.

Screening of antimicrobial potential of polysaccharide from cuttlebone and methanolic extract from...

Documents

Transcript of Screening of antimicrobial potential of polysaccharide from cuttlebone and methanolic extract from...