KR20040094886A - Composition comprising zoanthus sp. extract with anti-fouling activity and a use thereof - Google Patents

Abstract

The present invention provides a composition useful as an antifouling agent by inhibiting microbial growth, including Joanthus spp. Extract and fractions thereof, and optionally deodorants, and the extracts and fractions having about 100% antibacterial activity against contaminating microorganisms. It is about a use.

Description

Compositions comprising Joanthus spp. Extracts having anti-pollution activity and uses thereof COMPOSITION COMPRISING ZOANTHUS SP. EXTRACT WITH ANTI-FOULING ACTIVITY AND A USE THEREOF}

Chemical anti-pollution defense strategies against marine organisms have been studied, either directly against target contaminating organisms such as bacteria, fungi, algae, invertebrate larvae, or the like (Amade et al., 1987, Mar. Biol. 94, 271-275; Slattery et al., 1995, J. Exp. Mar. Biol. Ecol, 190, 61-77; Shellenberger and Ross, 1998, Northwest Science, 72 (1), 23-33)], or settlement There are other means of contamination such as obstruction, larval removal rate, photosynthesis, etc. (Kon-ya et al., 1994, Fish. Sci, 60 (6), 773-775; Kato et al., 1995, Tetrahedron letters, 36 (12), 2133-2136; Mizobuchi et al., 1996, Fisheries Sci 62 (1), 98-100; Becerro et al., 1997, J. Chem. Ecol, 23 (6), 1527-1547).

Information on the production of secondary metabolites in marine organisms is largely about the isolation and characterization of compounds and little is known about their ecological and biological significance. Recently, attempts have been made in this direction to some groups, such as soft corals, gorgonian, algae, sponges, bryozoan [Targett, 1988, in Marine Biodeterioration, MF Thompson., R. Sarojini., R. Nagabhushanam., AA Balkema eds, Rotterdam, 607-617; Standing et al., 1984, J Chem. Ecol, 10, 823-824; Davis et al., 1991, Mar. Ecol, Prog Ser, 72, 117-123; Vitalina et al., 1991, in Bioactive compounds from marine organisms, MF Thompson., R. Sarojini., R. Nagabhushanam eds, Oxford and IBH Publishing Co. Pvt. Ltd, 63-68 Szewzyk et al., 1991, Mar. Ecol. Prog. Ser, 75, 259-265.] Many other groups have not been investigated. Zoanthid (Phylum Coelenterata; Family Zoanthidae) is one of the few groups that received little attention with regard to chemical defense strategies.

Joantids live under water, have no nipples or stems, and contain thousands of individual polyps. Several compounds with various pharmacological properties have been isolated from coantides (Moore and Scheuer, 1971, Science, 172, 495-498; Cimino et al., 1973; Sturaro et al., 1982; Rao et al. , 1985, J. Org.Chem, 50, 3757-3760; Bakus et al., 1986, J Chem. Ecol, 12, 951-987; Quinn, 1988, in Bioorganic Marine Chemistry, PJ Scheuer ed.Vol. 2, 25-27; See Shigemori et al., 1999, J Nat. Prods, 62 (2), 373). However, little is known about the function of these metabolites in their natural habitats. At the Anjuna beach of Goa (Indian West Coast), they occur as dense mats and colonize large intertidal regions. Thus, the main focus of the present invention is to identify one natural product that has anti-contamination defense properties against conventional micro-contaminant organisms, ie bacteria and diatoms, uses extracts of Joanthus species and is useful in anti-contamination techniques. It's about ujidjrjtjadj ^ e ^ h ^ nucal.

Surfaces exposed to the marine environment undergo sequential changes that begin with the adsorption of the bio-polymer layer, followed by the formation of bacterial films, the growth of pinnate diatoms and protozoa (Baier, 1984, in Marine Biodeterioration: An Interdisciplinary Study, JD Costlow and RC Tipper, ed, Naval Institute Press, Anapolis, Maryland, 57-62; Mitchell and Kirchman, 1984, in Marine Biodeterioration: An Interdisciplinary Study, JD Costlow and RC Tipper, ed, Naval Institute Press, Anapolis, Maryland, 49-56; Wahl, 1989, Mar. Ecol. Prog. Ser, 58, 175-189). Thus, the numerous bacterial species forming the microbial film may affect, inhibit or sometimes have no effect on subsequent settled species that make up invertebrate larvae that settle and become adult (Bakus et al., 1986). , J. Chem. Ecol, 12, 951-987). In most cases where microbial films have been found necessary for the settlement of larvae and algae propagules, inhibition of bacterial growth prevents microbial film formation and thus inhibits larval settlement and ultimately epibiiont the surface. Will be maintained without).

As part of this effort, test animals were collected and crude methanol extracts were prepared. The anti-fouling potential of the extract was then examined for contaminating diatoms and bacteria known in the laboratory. After testing the activity of the crude extract, it was isolated using increasing polar solvents. These fractions were tested for the contaminating diatoms. As a result of the evaluation, it was found that the extract of Joanthus species has anti-fouling properties.

Object of the present invention

The main object of the present invention is to develop a composition comprising an extract of Joanthus spp.

Another main object of the present invention is to develop a composition having anti-fouling properties.

It is another object of the present invention to develop fractions from extracts of animal Joanthus species.

Another object of the present invention is to fractionate the crude extract using solvents of increasing polarity, ie petroleum ether, chloroform, n-butanol and an aqueous fraction.

Another object of the present invention is to identify the most effective fractions for contaminating diatoms.

Another object of the present invention is to develop a use for preparing a composition comprising an extract of Joanthus spp.

Another object of the present invention is to develop a use for the preparation of the composition having 100% anti-bacterial activity.

Another object of the present invention is to develop a use for preparing the composition having 100% microbial growth inhibitory activity against contaminating microorganisms.

It is another object of the present invention to develop a use for the preparation of such compositions having anti-microbial activity which is particularly effective in marine environments.

It is another object of the present invention to develop a use for the preparation of such compositions having particularly effective anti-microbial activity against bacteria and diatoms.

Summary of the invention

The present invention relates to compositions comprising extracts and / or fractions thereof from Joanthus species, and optionally deodorants, which are useful as anti-fouling agents by inhibiting microbial growth, and about 100% antibacterial active contaminating microorganisms. It relates to the use of the extract and its fractions having the same.

The present invention relates to compositions comprising extracts and / or fractions thereof from Joanthus species, and optionally deodorants, which are useful as anti-fouling agents by inhibiting microbial growth, and about 100% antibacterial active contaminating microorganisms. It relates to the use of the extract and its fractions having the same.

Accordingly, the present invention provides a composition comprising extracts and / or fractions thereof from Joanthus species, and optionally deodorants, useful as anti-fouling agents by inhibiting microbial growth, and about 100% antibacterial activity. It relates to the use of said extract and its fractions with contaminating microorganisms.

In one embodiment of the present invention there is provided a composition comprising an extract from Joanthus species and / or fractions thereof, and optionally a deodorant, useful as an anti-fouling agent against microorganisms.

In another embodiment of the invention, the extract and / or fractions thereof are prepared using polar or nonpolar solvents.

In another embodiment of the present invention, the preferred animal species to be used for said activity is Coelenterata Zoanthidea.

In another embodiment of the invention, the composition is prepared using systemic tissue of the species. In another embodiment of the present invention, the de-odorizing agent has no effect on the activity of the extracts and fractions.

In another embodiment of the present invention, the ratio of extract and / or fraction and deodorant is 1: 1 to 1:10.

In another embodiment of the invention, the composition is particularly effective against bacteria and diatoms.

In another embodiment of the invention, the invention relates to the use of producing extracts and fractions from Zoanthus sp useful as antifouling agents by inhibiting microorganisms.

In another embodiment of the invention, the washed sample of the species is immersed in methanol.

In another embodiment of the invention, crude extracts are obtained from systemic tissues of the species.

In another embodiment of the invention, the solvent extract is decanted.

In another embodiment of the invention, the decanted extract is filtered.

In another embodiment of the invention, the filtrate is evaporated at 40 ° C. to obtain a concentrated crude extract.

In another embodiment of the present invention, the concentrated crude extract is classified using an increased polar solvent.

In another embodiment of the invention, the crude extract is introduced into sterile filter paper discs at a concentration in the range of 0.1-5.0 mg / 6 mm diameter discs in methanol.

In another embodiment of the invention, the solvent is evaporated from the disc.

In another embodiment of the invention, the microorganism is exposed to the extract.

In another embodiment of the invention, the disc is placed in a medium plate supplied with test bacteria.

In another embodiment of the invention, the control is performed with a solvent that does not contain an extract.

In another embodiment of the invention, the plate is incubated overnight at a temperature of 25 to 32 ° C.

In another embodiment of the invention, the growth inhibition region is surveyed.

In another embodiment of the invention, the extract is used to determine the inhibition rate of microorganisms.

In another embodiment of the invention, diatoms are placed at an initial cell suspension concentration of 100-5000 cells / ml in the flask to determine the antidiatom activity of the crude extract and the fractions.

In another embodiment of the invention, the crude extract at a concentration of 10-5000 ppm is added to the cell suspension.

In another embodiment of the invention, the fraction is added in a concentration of 10-500 ppm in a separate flask.

In another embodiment of the present invention, the flask is incubated at a temperature of 25-32 ° C. under 12 hours of light and then 12 hours of dark exposure.

In another embodiment of the present invention, growth inhibition is determined after about 6 days, ie during the exponential growth phase of diatoms.

In another embodiment of the invention, said species preferably used for said activity is Coelenterata Zoanthidea .

In another embodiment of the invention, said use is particularly effective in marine environments.

In another embodiment of the invention, the use is particularly effective against bacteria and diatoms.

In another embodiment of the invention, the washed sample is immersed in methanol for 5 to 15 days.

In another embodiment of the present invention, the species is obtained from the rocks of the intertidal coast.

In another embodiment of the invention, the decanted extract is filtered through Whatman filter paper I.

In another embodiment of the invention, the filtrate is evaporated in vacuo.

In another embodiment of the present invention, the use uses Zobel marine media for bacterial culture.

In another embodiment of the invention, the concentration of said extract for bacteria is preferably about 0.5 mg / mm diameter disk.

In another embodiment of the invention, the extract is preferably incubated at 28 ° C.

In another embodiment of the invention, growth inhibition regions of less than 1 mm are recorded.

In another embodiment of the invention, the fractionation solvent is selected from the group comprising petroleum ether, chloroform, n-butanol and aqueous solutions.

In another embodiment of the invention, the nonpolar fractional petroleum ether is dissolved in acetone (0.005-.01 ml).

In another embodiment of the invention, the bacterium is Bacillus cereus, B.. B. circulans, Pseudomonas vesicularis, and blood. P. Putida.

In another embodiment of the invention, the antibacterial activity of the extract is 100%.

In another embodiment of the present invention, diatoms are selected from the group comprising Navicula subinflata and Nitzschia closterium.

In another embodiment of the invention, the calculation of growth inhibition with extracts and / or fractions with 50% growth inhibition is considered as the active source.

In another embodiment of the present invention, the extract exhibits 100% inhibition rate in diatoms at a concentration of 350 ppm or more.

In another embodiment of the invention, all said fractions of the crude extract exhibit inhibitory activity against diatoms.

In another embodiment of the invention, the petroleum ether fraction is the most effective of all fractions for diatoms.

In another embodiment of the invention, the petroleum ether fraction exhibits 100% inhibition against diatoms at a concentration of about 80 ppm or more.

In another embodiment of the present invention, the chloroform fraction shows about 100% inhibition against diatoms at a concentration of about 200 ppm.

In another embodiment of the present invention, the n-butanol fraction shows about 100% inhibition against diatoms at a concentration of 150-250 ppm.

In another embodiment of the present invention, the aqueous fraction shows 70-85% inhibitory activity against diatoms at a concentration of 140-250 ppm.

In another embodiment of the invention, the extract / fraction is free of toxicity / side effects on non-target organics.

In another embodiment of the invention, media plate diffusion is used to analyze the microorganisms in the medium.

In another embodiment of the invention, the use is used to prevent contamination in food.

In another embodiment of the invention, the use is used to prevent contamination in biological samples.

In another embodiment of the invention, the invention is

i) collecting live / fresh samples of Zoanthus sp from rocks on the intertidal shores,

ii) washing the sample cleanly with fresh sea water and soaking in methanol for about 10 days,

iii) filtering the extract with Whatman filter paper I and concentrating to a residue prepared by vacuum evaporation at 40 ° C.

It provides a method of preparing a crude extract from Zoanthus sp, comprising.

In one embodiment, the crude extract of Zoanthus sp exhibits antibacterial properties.

In a preferred embodiment, the crude extract of Zoanthus sp is Bacillus cereus , Bacillus circulans , Bacillus pumilus , Pseudomonas vesiciulans and Pseudomonas vesiciulans . Inhibits the growth of contaminating bacteria, such as Pseudomonas putida .

In another embodiment, the crude extract of Zoanthus sp inhibits the growth of contaminated diatoms at all concentrations.

In a preferred embodiment, crude extracts of Joanthus species are grown at a concentration of at least 600 ppm of diatom Navicula subinflata , at a concentration of at least 400 ppm of Nitzshia closterium and at least 400 ppm of Nitzshia closterium . I suppress it.

In another embodiment of the present invention, a fraction of crude antifouling extract from Joanthus species, based on increasing solvent polarity, ie, based on petroleum ether, chloroform, n-butanol, and aqueous fraction, and A process for testing the antifouling properties of contaminated diatoms of Navikula Subinplata and Nizshua Closterium (this process involves obtaining crude extracts from live and freshly collected Joantus species, Washing with water, immersing in methanol for about 10 days, filtering the extract through Whatman filter paper I, and concentrating the crude residue under vacuum at 40 ° C.). Presented.

In one embodiment, the crude extract of Joanthus species is fractionated based on increasing the polarity of the solvent with petroleum ether, chloroform, n-butanol and aqueous fractions.

In another embodiment, a total of four fractions of Joanthus species inhibit the growth of contaminating diatoms Nabikula Subinplata and Nizshua Closterium.

In another embodiment, the percentage inhibition in all fractions varies with concentration.

In another embodiment, petroleum ether fractions of Joanthus species are more effective at inhibiting the growth of contaminating diatoms Nabicula suvinplata and Nizshua closterium.

In a further embodiment, the petroleum ether fraction of the crude extract of Joanthus spp. Completely inhibits the growth of Navikula suvinplata at a concentration of at least 160 ppm, Nizchia Closterium at a concentration of at least 80 ppm.

In a further embodiment of the present invention, a method for preparing antifouling crude extracts from Joanthus species and testing the crude extracts for contaminating bacteria and diatoms is provided. The present invention also examines the method for fractionating crude extracts and the antifouling properties against contaminating bacteria of the fractions.

The method for producing an antifouling crude extract from Joanthus spp.

Collection of live / fresh samples of Joanthus species from intertidal rocks,

Wash samples with fresh sea water and soak in methanol for about 10 days,

Extractive filtration through Whatman filter paper I and vacuum distillation at 40 ° C. for concentration to crude residue.

It includes.

In another embodiment of the present invention, the crude extract of Joanthus species exhibits antimicrobial properties.

In another embodiment of the present invention, Joan tooth species crude extract is Bacillus cereus (Bacillus cereus), Bacillus CYR particulate lance (Bacillus circulans), Bacillus pumil Ruth (Bacillus pumilus), Pseudomonas shi Circular less (Pseudomonas vesiciularis) of and Inhibits the growth of contaminating bacteria, such as Pseudomonas putida .

In another embodiment of the present invention, crude extracts of Joanthus species inhibit the growth of contaminating diatoms at all concentrations.

In another embodiment of the present invention, the crude extract of Joanthus spp. Completely inhibits its growth at a concentration of 600 ppm or higher of diatoms Nabikula suvinplata and at a concentration of 400 ppm or higher of Nischia Closterium.

In another embodiment of the present invention, crude extracts of Joanthus species may have potential use as antifouling agents in aquatic environments.

In another embodiment of the invention, a method for fractionating crude antifouling extracts from Joanthus species based on increasing solvent polarity, ie petroleum ether, chloroform, n-butanol and aqueous fractions. Obtaining crude extract from freshly harvested Joanthus species, washing it cleanly with seawater, immersing in methanol for about 10 days, filtering the extract through Whatman filter paper I, and drying under vacuum at 40 ° C. Concentrating to the resulting residue).

In another embodiment of the present invention, crude extracts of Joanthus species are fractionated based on increasing the polarity of the solvent with petroleum ether, chloroform, n-butanol and aqueous fractions.

In another embodiment of the present invention, a total of four fractions of crude extracts of Joanthus species inhibit the growth of contaminating diatoms nabicula suvinplata and nizchia closterium.

In another embodiment of the invention, the percentage inhibition in all fractions varies with concentration.

In another embodiment of the present invention, the petroleum ether fraction of the crude extract of Joanthus species is more effective at inhibiting the growth of contaminating diatoms nabicula suvinplata and Nizshua closterium.

In another embodiment of the present invention, the petroleum ether fraction of the crude extract of Joanthus species inhibits its growth completely at a concentration of at least 160 ppm of Navikula subvinplata and at a concentration of at least 80 ppm of Nizchia Closterium.

In another embodiment of the invention, all fractions of crude extracts of Joanthus species, in particular petroleum ether fractions, may have potential use as antifouling agents in aquatic environments.

The invention is further illustrated by the following examples and should not be construed as limiting the scope of the invention.

Example 1

Joanthus species collected from Anjuna Beach (Goa) were used for the present invention. During the low tide, these animals were carefully scraped from intertidal rocks using metal spatula. A sample weight of about 2 kg was used for this study. Samples were washed thoroughly with fresh sea water and soaked in methanol for 6-10 days to prepare the crude extract. Extracts were obtained from whole body tissues. The solvent extract was decanted, filtered through Whatman filter paper I, and evaporated in vacuo at 40 ° C. to give a concentrate to the crude extract. About 30 gm of crude extract was thus obtained. This example illustrates the methodology used for the preparation of crude methanol extracts from Joanthus species.

Example 2

Five bacterial isolates, all belonging to the contaminated community, were used for the analysis. Bacterial analysis was performed by agar plate diffusion method (Amade et al., 1987, Mar. Biol, 94, 271-275) using Zobell Marine Agar (Himedia, Mumbai). Bacterial strains used in the analysis were Bacillus cereus, B. cyculus, B. pumilus, Pseudomonas veciculis and P. putida. A concentrate of 0.5 mg / 6 mm diameter disc of extract prepared in methanol was introduced onto the sterile filter paper disc. After solvent evaporation, the plates were placed on agar plates sprayed with test bacterial control (duplication replicated) containing only solvent. Four replicates of each concentration were used for the analysis and averaged. Plates were incubated overnight at room temperature (<28 ° C.). Growth inhibition zone was measured in mm (Berquist and Bedford, 1978, Mar. Biol , 46, 215-221). Inhibition zones measured below 1 mm were not scored. The results of this study are provided in Table 1. This example illustrates a methodology for performing antimicrobial assays using extracts of Joanthus spp.

Antibacterial Screening with Crude Extracts of Joanthus Species Germ control Bacillus cereus + Bacillus sirculans + Bacillus pumilus + Pseudomonas Besicularis + Pseudomonas Putida + + = 1-2 mm diameter suppression

Example 3

Both diatom Navicula subinflata and Nitzschia closterium belong to contaminated populations and were maintained in flotation water (Gentile and Johnsor, 1974, in, Proceedings on a Workshop on Marine Bioassays) , convened by Geraldine Cox, USA, 127). Actively growing cultures with an initial cell suspension of about 500-1000 cells ml " ¹ were used for analysis. The concentration series of extracts ranged from 200 to 1000 ppm. Aliquots of each extract stock solution prepared in distilled water The amount (1 cc = 50 mg) was taken according to the desired experimental concentration and the volume was adjusted to 50 ml in a 125 ml Elenmeyer flask All analyzes were performed four times The control flask was only algae cells in medium The flasks were incubated at room temperature (about 28 ° C.) for 12 hours of light and 12 hours of dark exposure conditions, growth inhibition was after 6 days, ie algae attached using Sedgwick rafter. Determined during exponential growth phase by number of cells and expressed as% growth inhibition in Table 2. Extracts that can provide 50% growth inhibition were considered active. Not to carry out tooth species / anti diatoms analysis / Using illustrates the methodology used.

Inhibition of growth of diatoms at different concentrations of crude extracts of Joanthus spp. Diatom species Concentration (ppm) % Suppression ± SD Navikula Suflata 200 49.7 ± .8 400 96.3 ± 0.4 600 100.0 ± 0.0 800 100.0 ± 0.0 1000 100.0 ± 0.0 Nitzshua Closterium 200 5.5 ± 8.3 400 100.0 ± 0.0 600 100.0 ± 0.0 800 100.0 ± 0.0 1000 100.0 ± 0.0

Example 4

The crude extract was then fractionated into four fractions using a highly polar solvent, ie petroleum ether, chloroform, n-butanol and aqueous solution. All fractions were tested for activity as shown in Table 3 using the above-mentioned usage. o ￡. The nonpolar fraction insoluble in distilled water during the preparation of the stock solution was dissolved in acetone (0.005-0.01 ml). This set of control flasks contained cells + acetone (the same volume added to make a stock solution) in medium. This example illustrates the methodology used for the fractionation of crude extracts and their testing for anti-diagnostic analysis.

% Growth Inhibition of Diatoms at Different Concentrations of Four Fractions of Crude Extracts of Joanthus Species Diatom species Concentration (ppm) % Suppression ± SD Petroleum ether chloroform n-butanol Mercury Navikula Subinplata 40 22.8 ± 3.0 stimulus 2.8 ± 1.6 stimulus 80 30.5 ± 3.0 stimulus 6.7 ± 1.6 11.3 ± 4.2 120 99.7 ± 0.1 stimulus 15.0 ± 1.2 42.2 ± 9.3 160 100.0 ± 0.0 14.8 ± 5.4 64.6 ± 0.9 44.7 ± 21.4 200 100.0 ± 0.0 89.4 ± 11.7 74.5 ± 1.2 69.0 ± 7.1 Nitzshua Closterium 40 81.0 ± 5.2 stimulus 5.2 ± 6.8 23.3 ± 11.3 80 100.0 ± 0.0 44.1 ± 8.9 14.8 ± 1.6 30.7 ± 3.0 120 100.0 ± 0.0 50.4 ± 5.9 59.9 ± 7.2 48.7 ± 13.7 160 100.0 ± 0.0 77.7 ± 14.8 100.0 ± 0.0 67.2 ± 6.3 200 100.0 ± 0.0 100.0 ± 0.0 100.0 ± 0.0 77.8 ± 8.7

Example 5

This example describes the methodology used for the treatment of results during antidiatomical analysis with fractions of crude and crude extracts.

% Growth inhibition is calculated using the following formula.

Where inocoolum cell number at O = 0 hours; A = number of cells attached in control flasks after 6 days; B = number of cells attached in experimental flask after 6 days.

Advantage of the present invention

1. This is the first time to confirm the antifouling properties of crude extracts of Joanthus spp.

2. Laboratory studies have shown that crude extracts of Joanthus species effectively inhibit the growth of contaminating bacteria and diatoms, the major organisms that colonize any oceanic base.

3. All four fractions, ie petroleum ether, chloroform, n-butanol and aqueous fractions, were separated from crude extracts showing antifouling properties.

4. The petroleum ether fraction of the four fractions was found to be more effective at inhibiting the growth of contaminating algae.

5. This is a natural product that may not have toxicity / side effects on non-target organisms.

6. It has the potential to form natural compounds in antifouling products.

Claims (35)

A composition of matter comprising one of an petroleum ether fraction, a chloroform fraction, an n-butanol fraction and an extract fraction of Zoanthus sp. Which is an aqueous fraction. The composition of claim 1 wherein the fraction is prepared using a polar or nonpolar solvent. The composition of claim 1 wherein the preferred animal species used for said activity is Coelenterata Zoanthidea . The composition of claim 1 prepared using systemic tissue of said species. The composition of claim 1, which is particularly effective against bacteria and diatoms. (a) dipping a washed sample of Zoanthus sp. in methanol, (b) obtaining a crude extract from systemic tissues of said species, (c) decanting with solvent extracts, (d) filtering the decanted extract, (e) extracting the filtrate at 40 ° C. to obtain a concentrated crude extract, (f) sorting the concentrated crude extract with an increased polar solvent, (g) introducing the crude extract in a concentration ranging from 0.1 to 50 mg / 6 mm diameter disc into methanol or sterile filter paper disc, (h) evaporating the solvent from the disc, (i) exposing the microorganisms to the extract, (j) placing the disk on agar plates seeded with test bacteria, (k) performing control with a solvent free of extracts, (l) incubating the plate overnight at a temperature of 25-32 ° C., (m) measuring the growth inhibition zone, (n) determining the percentage of inhibition of the microorganism using the extract, (o) measuring the anti-diatom activity of the crude extract and the fraction separately by placing the diatoms in an initial cell suspension at a concentration of 100 to 5000 cells / ml in the flask, (p) adding the crude extract at a concentration of 10 to 5000 ppm to the cell suspension, (q) adding said fraction at a concentration of 10 to 500 ppm to a separate flask, (r) light incubating the flask at a temperature of 25 to 32 ° C. for 12 hours and then incubating for 12 hours under dark exposure conditions, (s) from Zoanthus sp. useful as an anti-founling agent after about 6 days, i.e. by measuring the percentage of growth inhibition during diatom exponential growth phase . For the preparation of extracts and fractions thereof. 9. Use according to claim 8, wherein the species preferably used for said activity is Coelenterata Zoanthidea. 9. Use according to claim 8, which is particularly effective in marine environments. 9. Use according to claim 8, which is particularly effective against contaminating microorganisms. 9. Use according to claim 8, which is particularly effective for bacteria and diatoms. Use according to claim 8, wherein the washed sample is immersed in methanol for 5 to 15 days. 9. Use according to claim 8, wherein said species are obtained from intertidal rocks. The use according to claim 8, wherein the decanted extract is filtered through whatman filtration pattern I. The use according to claim 8, wherein the filtrate is evaporated using a vacuum. The use according to claim 8, wherein Zobel marine agar is used for bacterial culture. The use according to claim 8, wherein the concentration of said extract relative to bacteria is preferably a diameter disk of about 0.5 mg / mm. The use according to claim 8, wherein said extract is incubated at 28 ° C. The use according to claim 8, wherein the growth inhibition zone of less than 1 mm is not recorded. The use according to claim 8, wherein the classification solvent is selected from the group comprising petroleum ether, chloroform, n-butanol and aqueous. The use according to claim 8, wherein the nonpolar fractional petroleum ether is dissolved in acetone (0.005 to 0.01 ml). The bacterium of claim 8, wherein the bacterium is Bacillus cereus, b. B. circulans, Pseudomonas vesicularis and blood. Use is selected from the group comprising p. Putida. Use according to claim 8, wherein the antibacterial activity of the extract is 100%. Use according to claim 8, wherein the diatoms are selected from the group comprising Navicula subinflata and Nitzschia closterium. 9. Use according to claim 8, wherein calculating the percentage growth inhibition with extracts and / or fractions having growth inhibition greater than 50% is considered active. The use according to claim 8, wherein said extract exhibits 100% inhibition in diatoms at a concentration of at least 350 ppm. 9. Use according to claim 8, wherein all of the crude extract fractions exhibit inhibitory activity against diatoms. The use according to claim 8, wherein the petroleum ether fraction is the most effective of all fractions for diatoms. The use according to claim 8, wherein the petroleum ether fraction exhibits 100% inhibition at diatoms at a concentration of at least about 80 ppm. The use according to claim 8, wherein the chloroform fraction shows about 100% inhibition in diatoms at a concentration of about 200 ppm. The use according to claim 8, wherein the n-butanol fraction shows about 100% inhibition against diatoms at a concentration of 150 to 250 ppm. The use according to claim 8, wherein the aqueous fraction exhibits 70-85% inhibitory activity against diatoms at a concentration of 140-250 ppm. The use according to claim 8, wherein said extract / fraction does not show toxicity / side effects on non-target organisms. 9. The use of claim 8 for assaying microorganisms in culture medium using agar plate diffusion applications. 9. Use according to claim 8 for preventing contamination in foods using said use. The method of claim 8, wherein said use is used to prevent contamination in a biological sample.