Potential antimicrobial activity of marine molluscs from tuticorin, southeast coast of India against 40 biofilm bacteria.ABSTRACT Methanol methanol, methyl alcohol, or wood alcohol, CH3OH, a colorless, flammable liquid that is miscible with water in all proportions. Methanol is a monohydric alcohol. It melts at −97. : water (1:1), methanol: dichloromethane (1:1) and acetone acetone (ăs`ĭtōn), dimethyl ketone (dīmĕth`əl kē`tōn), or 2-propanone (prō`pənōn), CH3COCH3 extracts of molluscs comprising 77 whole body, four inks, four opercula o·per·cu·lum n. pl. o·per·cu·la or o·per·cu·lums A lid or flap covering an aperture, such as the gill cover in some fishes or the horny shell cover in snails or other mollusks. , 10 egg masses, and 10 digestive glands digestive gland n. A gland, such as the liver or pancreas, that secretes into the alimentary canal substances necessary for digestion. were screened for antimicrobial antimicrobial /an·ti·mi·cro·bi·al/ (-mi-kro´be-al) 1. killing microorganisms or suppressing their multiplication or growth. 2. an agent with such effects. activity on marine biofilm Biofilm An adhesive substance, the glycocalyx, and the bacterial community which it envelops at the interface of a liquid and a surface. When a liquid is in contact with an inert surface, any bacteria within the liquid are attracted to the surface and adhere bacteria. The methanol: water (1:1) whole body extracts of Nerita albicilla and Nerita oryzarum showed broad spectral spectral /spec·tral/ (spek´tral) pertaining to a spectrum; performed by means of a spectrum. spec·tral adj. Of, relating to, or produced by a spectrum. inhibitory activity against 93% and 95% of the 40 biofilm bacteria. The egg masses from 10 gastropods showed activity against more than a quarter of the biofilm bacteria. The methanolic extract of Chicoreus virgineus, Chicoreus ramosus egg masses, and acetone extract of the egg mass of Rapana rapiformis showed broad-spectrum antibacterial antibacterial /an·ti·bac·te·ri·al/ (-bak-ter´e-al) destroying or suppressing growth or reproduction of bacteria; also, an agent that does this. an·ti·bac·te·ri·al adj. activity against all the 40 biofilm bacterial strains. The activity in gastropod gastropod, member of the class Gastropoda, the largest and most successful class of mollusks (phylum Mollusca), containing over 35,000 living species and 15,000 fossil forms. egg masses was localized to their internal matrix. Overall screening showed activity in 38.1% of the methanolic extracts followed by 13.3% of methanol: water, 12.4% of methanol: dichloromethane, and 3.8% of acetone extracts. Gastropods showed good activity when compared with bivalves and cephalopods. KEY WORDS: antifouling an·ti·foul·ing adj. Counteracting or preventing the building up of deposits on underwater surfaces, such as the undersides of boats: antifouling paint. , bacteria, biofilm, egg mass, mollusc mollusc members of the phylum Mollusca, which comprises about 50,000 species. Includes snails, slugs and the aquatic molluscs—oysters, mussels, clams, cockles, arkshells, scallop, abalone, cuttlefish, squid. INTRODUCTION The efforts to control marine biofouling bi·o·foul·ing n. The impairment or degradation of something, such as a ship's hull or mechanical equipment, as a result of the growth or activity of living organisms. are ongoing since humans started venturing into the sea. Among the antifoulants developed to date, organotin compounds are considered as the most effective and have been widely used since the 1970s. It is well established that TBT TBT, n See theta brainwave training. TBT Transcervical balloon tuboplasty, see there compounds cause imposex universally (Barroso et al. 2002) and has been documented in over 118 species in 63 genera genera, in taxonomy: see classification. (Bettin et al. 1996). Organotin pollution has also raised concerns over the accumulation of organotin compounds in the food chain and the associated risks related to the presence of contaminants (Belfroid et al. 2000). International Maritime Organization International Maritime Organization (IMO), specialized agency of the United Nations established in 1948, with headquarters in London and 158 member nations. IMO is one of the smallest of the UN agencies. (IMO "In my opinion." See IMHO and digispeak. IMO - IMHO ) has proposed a complete phasing out of TBT use by the year 2008. The alternative tin free coatings are not only costly, but also their metal constituents may pose environmental problems (Hellio et al. 2004). So, a great deal of research has been focused on finding an alternate nontoxic and eco-friendly antifoulant (De Nys & Steinburg 2002, Fusetani 2004). In this context, marine organisms, especially the sedentary sedentary /sed·en·tary/ (sed´en-tar?e) 1. sitting habitually; of inactive habits. 2. pertaining to a sitting posture. sedentary of inactive habits; pertaining to a fat, castrated or confined animal. forms have interested researchers very much because they exhibit characteristic chemical defense against the epizooic organisms. More than 100 species of marine organisms have been shown to exhibit antimicrobial activity, as well as the ability to prevent settlement of fouling organisms (Clare 1996, Koh 1997). Most currently available antifouling compounds are active against barnacles and algae algae (ăl`jē) [plural of Lat. alga=seaweed], a large and diverse group of primarily aquatic plantlike organisms. These organisms were previously classified as a primitive subkingdom of the plant kingdom, the thallophytes (plants that and not against microbes like biofilm bacteria, which cause initial fouling of a submerged surface (Tang tang, in zoology tang: see butterfly fish. & Cooney 1998). Bacteria are the first organisms to foul submerged surfaces in the marine environment. Their subsequent multiplication multiplication, fundamental operation in arithmetic and algebra. Multiplication by a whole number can be interpreted as successive addition. For example, a number N multiplied by 3 is N + N + N. and production of exopolymers leads to the formation of bacterial biofilms, which attract a large number of secondary fouling organisms (Steinberg et al. 2002). The interaction between bacteria and macrofoulers are dynamic and varied. The resultant biofouling community leads to economic and ecologic implications on both human-made structures and living surfaces. However, bacteria do not colonize col·o·nize v. col·o·nized, col·o·niz·ing, col·o·niz·es v.tr. 1. To form or establish a colony or colonies in. 2. To migrate to and settle in; occupy as a colony. 3. all submerged surfaces in a uniform manner and are influenced by many physical and chemical factors (Fletcher & Marshall 1982). The colonization colonization, extension of political and economic control over an area by a state whose nationals have occupied the area and usually possess organizational or technological superiority over the native population. process of the bacteria in the living marine surfaces may also be influenced by the secondary metabolites Secondary metabolites are organic compounds that are not directly involved in the normal growth, development or reproduction of organisms. Unlike primary metabolites, absence of secondary metabolities results not in immediate death, but in long-term impairment of the organism's produced by the epibiont host. There exists an inverse correlation between macrofouling and antimicrobial activity of the epibiont host (Koh 1997, Santhana Ramasamy & Murugan 2003). This has been proposed as evidence that secondary metabolites are involved in the control of epibiosis (Slattery et al. 1995, Newbold et al. 1999). So, the natural mechanism of upsetting the development of the microfouling bacterial community by nontoxic marine natural compounds produced by the foul free host organisms may be taken as the most desirable way for breaking the fouling chain to reduce macrofouling settlement. The cnidarians, sponges, ascidians, bryozoans, marine plant, and the like have received much attention and widely studied for antifouling activity (Clare 1996, Fusetani 2004). The study on the bioactivity bi·o·ac·tiv·i·ty n. The effect of a given agent, such as a vaccine, upon a living organism or on living tissue. of marine mollusc metabolites Metabolites Substances produced by metabolism or by a metabolic process. Mentioned in: Interactions is scanty except for the nudibranchs. Rinehart et al. (1981) opined that marine molluscs might provide potential for isolating compounds with specific activity against certain microorganisms or cell types. Molluscan mol·lus·can also mol·lus·kan adj. Of or relating to the mollusks. n. A mollusk. forms do exhibit adaptations to avoid fouling of their surface. The rare incidence of fouling on the Helgoland snails of Littorina littorea is due to the density dependent mutual grazing grazing, n See irregular feeding. grazing 1. actions of herbivorous animals eating growing pasture or cereal crop. 2. area of pasture or cereal crop to be used as standing feed. See also pasture. (Wahl et al. 1998). The periostracum per·i·os·tra·cum n. pl. per·i·os·tra·ca The hard chitinous outer covering of the shell of many mollusks, especially freshwater ones, that protects the shell from the erosive action of water. of the mussel mussel, edible freshwater or marine bivalve mollusk. Mussels are able to move slowly by means of the muscular foot. They feed and breathe by filtering water through extensible tubes called siphons; a large mussel filters 10 gal (38 liters) of water per day. Mytilus edulis has been shown to possess some repellents to avoid fouling of their surface (Wahl et al. 1998). The gelatinous gelatinous /ge·lat·i·nous/ (je-lat´i-nus) like jelly or softened gelatin. ge·lat·i·nous adj. 1. Of, relating to, or containing gelatin. 2. Resembling gelatin; viscous. egg masses of marine molluscs also have no physical protection from predation predation Form of food getting in which one animal, the predator, eats an animal of another species, the prey, immediately after killing it or, in some cases, while it is still alive. Most predators are generalists; they eat a variety of prey species. . So, many marine molluscs might have evolved mechanisms to avoid predation and surface fouling on their egg masses. Lord (1986) has demonstrated that the content of the egg capsules in Nucella lapillus la·pil·lus n. pl. la·pil·li A small, solidified fragment of lava. [Latin, diminutive of lapis, stone. was free from bacterial contamination and the capsule capsule In botany, a dry fruit that opens when ripe. It splits from top to bottom into separate segments known as valves, as in the iris, or forms pores at the top (e.g., poppy), or splits around the circumference, with the top falling off (e.g., pigweed and plantain). wall was impermeable impermeable /im·per·me·a·ble/ (-per´me-ah-b'l) not permitting passage, as of fluid. im·per·me·a·ble adj. Impossible to permeate; not permitting passage. to bacteria. Benkendorff et al. (2001) indicated that antibacterial compounds are common in both tough egg capsules and gelatinous egg masses, suggesting that chemical defense is the primary mechanism used to maintain an axenic axenic /axen·ic/ (a-zen´ik) not contaminated by or associated with any foreign organisms; used in reference to pure cultures of microorganisms or to germ-free animals. Cf. gnotobiotic. environment for embryonic development. The egg masses of some marine molluscs have been the source of several compounds with antibacterial activity (Kamiya et al. 1984, Matsunaga et al. 1986, Yamazaki 1993, Benkendorff et al. 2000a, Pawlik et al. 1988). But, study on the antimicrobial properties of the marine molluscs against marine bacteria is scanty except a few works (Kamiya et al. 1984, Benkendorff et al. 2000b, Mitta et al. 2000, Benkendorff et al. 2001) and no previous studies have tested for activity against biofilm bacteria. So, in this study antimicrobial activity of a broad range of marine molluscs was evaluated against marine biofilm bacteria. MATERIALS AND METHODS The screened molluscs in the present study are constituted by 77% gastropods, 21% bivalves, and 2% cephalopods comprising the whole body, digestive gland, opercula, ink, and egg masses. It is to be noted that the ecologic role of the activity has not been addressed in this study. Because the study is on shelled molluscs and only the inanimate inanimate /in·an·i·mate/ (-an´im-it) 1. without life. 2. lacking in animation. in·an·i·mate adj. surfaces of the animals are exposed to the marine environment, the whole body was selected instead of surface layer. The digestive gland was also analyzed from a subset of species, because there have been reports of bioaccumulation bi·o·ac·cu·mu·la·tion n. The increase in the concentration of a substance, especially a contaminant, in an organism or in the food chain over time. of secondary metabolites by some molluscs from their food (Faulkner 1988, Cimino & Sodano 1993, Rogers et al. 2000). The foul free surface of the opercula is the reason for its inclusion in the study. The previous report of antimicrobial property of egg masses (Benkendorff et al. 2001) has formed the basis of their selection. Extraction of Molluscs Molluscs collected alive from the fish landings of the Tuticorin coast (Lat (Local Area Transport) A communications protocol from Digital for controlling terminal traffic in a DECnet environment. LAT - Local Area Transport . 8[degrees]45'N; Long. 78[degrees]10'E) were transported to the laboratory and identified up to species level using standard keys (Satyamurti 1952a and b, Hylleberg & Kilburn 2002, Personal communications from Dr. Alan Kohn, University of Washington, USA and Mr. Peter Middelfart of Australian Museum This article is about the museum in Sydney. For the museum in Canberra opened in 2001, see National Museum of Australia. The Australian Museum is the oldest museum in Australia, with an international reputation in the fields of natural history and anthropology. , Australia; BIOSIS BIOSIS Biosciences Information Service Indo-Pacific Mollusc database). The shells were cracked by a light tap with a hammer to remove the soft parts. Then the tissues were rinsed with sterile distilled water Noun 1. distilled water - water that has been purified by distillation H2O, water - binary compound that occurs at room temperature as a clear colorless odorless tasteless liquid; freezes into ice below 0 degrees centigrade and boils above 100 degrees centigrade; and the whole body, digestive gland, opercula, inks, and egg masses of the respective molluscs were dissected dis·sect·ed adj. 1. Botany Divided into many deep, narrow segments: dissected leaves. 2. Geology Cut by irregular valleys and hills. Adj. 1. and extracted (10 mL solvent to 0.1 g of tissue) by using the solvents methanol: dichloromethane (1:1), methanol, methanol: water (1:1), and acetone separately. The solvents/combinations used for extraction were developed based on random experiments. Replicate tissue samples were used for extraction in different solvents. The molluscan whole body and the digestive glands were cut into small pieces and air-dried for 24 h at room temperature before extraction with solvents. The opercula were air dried, ground well, and extracted with solvents. Fresh encapsulated encapsulated Localized Oncology adjective Confined to a specific area, surrounded by a thin layer of fibrous tissue; encapsulation generally refers to a tumor confined to a specific area, surrounded by a capsule. See Islet encapsulation. egg masses were crushed before extraction with solvents. The developmental stages were not taken into consideration in this study. The effect of epibiotic bacteria and the presence of activity in the internal matrix of the egg masses of Chicoreus virgineus, C. ramosus, and Rapana rapiformis were determined by screening the empty (spent) egg masses. Raw inks obtained from the respective glands were directly used for extraction. The extracts were cold steeped overnight at -18[degrees]C, filtered with Whatman No. 1 filter paper, evaporated evaporated reduced in volume by evaporation; concentrated to a denser form. , and concentrated (Becerro et al. 1994, Riguera 1997, Wright 1998, Murugan & Santhana Ramasamy 2003). These crude extracts were used for antibacterial assay against biofilm bacterial strains isolated from the panels. Isolation of Biofilm Bacteria The fouling bacteria used in the antibacterial assay were isolated from the biofilm formed over wood, stainless steel stainless steel: see steel. stainless steel Any of a family of alloy steels usually containing 10–30% chromium. The presence of chromium, together with low carbon content, gives remarkable resistance to corrosion and heat. , and fiber panels by pour plate technique (Wahl 1995). The panels were deployed for about a month during February and June 2002 at a depth of 1 m in the vicinity of Tuticorin Harbour (Lat 8[degrees]45'N; Long 78[degrees]10'E). The panels were washed with sterile seawater seawater Water that makes up the oceans and seas. Seawater is a complex mixture of 96.5% water, 2.5% salts, and small amounts of other substances. Much of the world's magnesium is recovered from seawater, as are large quantities of bromine. before swabbing with sterile cotton swabs "Q-Tip" redirects here. For the rapper, see Q-Tip (rapper). For the band, see Q-Tips (band). Cotton swabs (British English: cotton buds) are used in first aid, cosmetics application, and a variety of other uses. . The swabs were placed in a tube containing sterile seawater, serially diluted and inoculated in marine agar plates An agar plate is a sterile Petri dish that contains a growth medium (typically agar plus nutrients) used to culture microorganisms. Selective growth compounds may also be added to the media, such as antibiotics. (Zobell Marine Agar, Himedia, Mumbai). The plates were incubated at room temperature for 7 days. The pure bacterial strains based on colony morphology morphology In biology, the study of the size, shape, and structure of organisms in relation to some principle or generalization. Whereas anatomy describes the structure of organisms, morphology explains the shapes and arrangement of parts of organisms in terms of such , pigmentation pigmentation, name for the coloring matter found in certain plant and animal cells and for the color produced thereby. Pigmentation occurs in nearly all living organisms. , and appearance were isolated by repeated streaking and identified up to genus genus, in taxonomy: see classification. genus Biological classification. It ranks below family and above species, consisting of structurally or phylogenetically (see level (Holt et al. 1994). The isolated strains were stored in marine agar slants at 4[degrees]C. Antibacterial Assay Antibacterial assay was carried out by using the standard disc diffusion method (Becerro et al. 1994, Slattery et al. 1995, Murugan & Santhana Ramasamy 2003). The disc diffusion assay with the marine bacteria as the target organism has been widely used as a primary screening tool to assess the antifouling potential of a substance (Clare 1996). The advantage of such a preliminary screening is that it is simple, less time-consuming, and requires only a small quantity of the material, which ensures minimal removal of biodiversity biodiversity: see biological diversity. biodiversity Quantity of plant and animal species found in a given environment. Sometimes habitat diversity (the variety of places where organisms live) and genetic diversity (the variety of traits expressed from the environment (Devi et al. 1997). But, the results of the disc diffusion assay have to be interpreted carefully taking into account the fact that the lipophilic lipophilic, adj/n the ability to dissolve or attach to lipids. lipophilic (lipōfil´ik), adj 1. showing a marked attraction to, or solubility in, lipids. 2. active compounds do not migrate across the watery wa·ter·y adj. 1. Filled with, consisting of, or soaked with water; wet or soggy. 2. Secreting or discharging water or watery fluid, especially as a symptom of disease. agar matrix (Benkendorff et al. 2000b). The disc diffusion assay in the present study is not used for quantitative estimation of the activity or for comparison between different species. The Whatman No. 1 sterilized ster·il·ize tr.v. ster·il·ized, ster·il·iz·ing, ster·il·iz·es 1. To make free from live bacteria or other microorganisms. 2. discs of 9-mm diameter, impregnated im·preg·nate tr.v. im·preg·nat·ed, im·preg·nat·ing, im·preg·nates 1. To make pregnant; inseminate. 2. To fertilize (an ovum, for example). 3. with 50 [micro]g/disc of the crude extract, and air dried were placed on the marine agar plates (90 x 15 mm) seeded with individual biofilm bacterial strains. The plates were incubated for 24 h at 30[degrees]C. The 40 biofilm bacterial strains isolated from the fouling panels were used for the antibacterial assay. The biofilm bacterial strains were grown in Zobell marine broth broth liquid media for culturing microorganisms. cooked meat broth a medium useful for culturing anaerobic bacteria. enrichment broth one modified to permit growth by selected bacteria. for 24 h before seeding the marine agar plates. The assay was carried out in triplicate. Control plates with solvents were also maintained separately. The zone of inhibition was measured from the edge of the disc to the clear zone in millimeter. The positive results of the antibacterial assay were expressed as percent inhibition of extract against 40 biofilm bacteria. RESULTS The 105 crude extracts of various parts of marine molluscs tested against 40 biofilm bacteria showed varied antibacterial activity (Tables 1, 2, 3, 4, and 5). The overall screening showed activity in 38.1% of the methanolic extracts followed by 13.3% of methanol: water, 12.4% of methanol: dichloromethane and 3.8% of acetone extracts. No activity against any biofilm bacteria was observed for 32.4% of the extracts. Characterization of Bacteria The 40 biofilm bacterial strains isolated from the fouling panels included five strains of Vibrio vibrio Any of a group of aquatic, comma-shaped bacteria in the family Vibrionaceae. Some species cause serious diseases in humans and other animals. They are gram-negative (see sp., four strains each of Micrococcus micrococcus Any of the spherical bacteria that make up the genus Micrococcus. Widespread in nature, these gram-positive (see gram stain) cocci (see coccus) are usually not considered to cause disease. sp., Pseudomonas Pseudomonas A genus of gram-negative, nonsporeforming, rod-shaped bacteria. Motile species possess polar flagella. They are strictly aerobic, but some members do respire anaerobically in the presence of nitrate. sp., Flavobacterium sp., Aeromonas sp., Bacillus bacillus (bəsĭl`əs), any rod-shaped bacterium or, more particularly, a rod-shaped bacterium of the genus Bacillus. Some bacterium in the genus cause disease, for example B. sp., Enterobacter sp., Alteromonas sp., Cornebacterium sp., and three species of Alcaligenes sp. (Table 6). Antibacterial Assay of Whole Body Extracts Nearly 71.4% of the 77 species tested using whole body molluscan extracts showed activity (Table 1). The 14.3% of the species showed activity against more than 50% of biofilm bacteria. Extracts from only two species (2.6%) showed high activity and inhibited more than 90% biofilm bacteria. The remaining 28.6% of the whole body extracts did not show any activity. Among the whole body extracts of the gastropods screened, 70.7% showed activity against some biofilm bacteria and no activity was observed in 29.3% of the species. Among the gastropod extracts, which showed activity, 75.6% showed inhibitory activity against at least 50% of biofilm bacteria and 24.4% inhibited the growth of more than 50% biofilm bacteria. The methanol: water (1:1) extracts of Strombus marginatus (23), methanol: dichloromethane (1:1) extract of Cronia margariticola (28), methanol extracts of Bufonaria rana (47), Turbo bruneus (7), and acetone extract of Murex mu·rex n. pl. mu·ri·ces or mu·rex·es Any of various marine gastropods of the genus Murex common in tropical seas and having rough spiny shells, especially M. trunculus, the source of Tyrian purple. tribulus (30) exhibited activity against more than 60% of bacteria (Table 1). The whole body methanol: water (1:1) extracts of Nerita albicilla and Nerita oryzarum showed broad spectral activity against 95% and 93% of the biofilm bacteria, and the inhibition zone ranged from nil to 4 mm and nil to 8 mm respectively (Table 6). N. oryzarum methanol: water extracts showed high inhibitory activity against Aeromonas sp. 1 (Table 6). With regard to the bivalves, 70.6% of the whole body extracts showed activity, almost equal to that of gastropod. But, no high activity (not more than 60% strains inhibited) was observed for bivalve bivalve, aquatic mollusk of the class Pelecypoda ("hatchet-foot") or Bivalvia, with a laterally compressed body and a shell consisting of two valves, or movable pieces, hinged by an elastic ligament. extracts. Nearly 91.7% of the bivalve species showed activity against less than 50% biofilm bacteria and 29.4% showed no activity. Among the bivalves, the methanol: dichloromethane extract of Pinna pinna /pin·na/ (pin´ah) auricle (1).pin´nal pin·na n. pl. pin·nae See auricle. pin bicolor bicolor a coat color of two colors. In dogs, usually black with tan markings but may be other combinations such as ticking on a white background. In cats, more than two spots of color on the body, either white and one basic color, or white with one tabby color. (72) showed high inhibitory activity against 55% biofilm bacteria. The activity of two cephalopod cephalopod (sĕf`ələpŏd'), member of the class Cephalopoda, the most highly organized group of mollusks (phylum Mollusca), and including the squids, octopuses, cuttlefish, and nautiluses. whole body extracts was not prominent and they exhibited inhibition of less than 40% of biofilm bacteria (Table 1). Antibacterial Assay of Digestive Gland, Ink, and Opercula Extracts The digestive gland extracts of the 10 gastropods did not show much activity. Only the methanol extracts of Melo melo (81) and Haliotis varia var·i·a n. A miscellany, especially of literary works. [Latin, from neuter pl. of varius, various.] (78) exhibited activity, with the remaining digestive gland extracts showing no activity (Table 2). The ink extracts of two gastropods and two cephalopods, screened for antibacterial activity, did not show promising inhibitory activity either. Only the methanolic extracts of Aplysia extraordinaria (88), Dolabella auricularia (89) inks showed activity, but against less than 50% of biofilm bacteria (Table 3). Among the five opercula screened, only that of Lambis lambis (93) and Chicoreus virgineus (95) showed activity (Table 4). The activity exhibited by opercula of Lambis lambis (93) was not prominent because it inhibited only 15% of biofilm bacteria. Antibacterial Assay of Molluscan Egg Mass Extracts Interestingly, all encapsulated egg masses of the 10 gastropods exhibited inhibitory activity (Table 5). Sixty percent of the egg masses showed activity against more than 50% of biofilm bacteria. The egg masses of the family Muricidae inhibited the highest number of biofilm bacteria, with, Chicoreus virgineus, C. ramosus, and Rapana rapiformis showing broad spectrum antibacterial activity against all the 40 biofilm bacterial strains (Table 6). The methanolic extract of Chicoreus virgineus egg mass showed an inhibition zone ranging from 1-10 mm against different bacteria. The acetone extract of the egg mass of Rapana rapiformis showed an inhibition zone of 1-7 mm. The methanolic extract of Chicoreus ramosus egg mass showed an inhibition zone of 1-5 mm. The extracts of these three egg masses showed high inhibitory activity against the biofilm bacteria Vibrio sp. 2 (Table 6). The extracts of empty egg masses did not show any activity. DISCUSSION Forty biofilm bacterial strains belonging to 10 genera were isolated and used in antibacterial assays. The strains are the common marine biofilm bacteria in India (Santhana Ramasamy & Murugan 2003). It is well known that many fouling invertebrates require the presence of a microbial microbial pertaining to or emanating from a microbe. microbial digestion the breakdown of organic material, especially feedstuffs, by microbial organisms. surface film as a prerequisite for settlement (Scheltema 1974). Many marine organisms produce bioactive bi·o·ac·tive adj. Of or relating to a substance that has an effect on living tissue. bioactive having an effect on or eliciting a response from living tissue. metabolites, which may play a direct role in preventing fouling (Pawlik 1993) and their production can be considered as a kind of autogenic au·tog·e·nous also au·to·gen·ic adj. 1. Produced from within; self-generating. 2. Medicine Originating with the individual to which applied: an autogenous graft; an autogenous vaccine. protection (Wahl et al. 1998). In this study, some marine molluscs were shown to inhibit the growth of biofilm bacteria, although since the activity was not associated with surface tissues, the ecologic role is uncertain. Nevertheless, the extracts of marine organisms showing antimicrobial activity against biofilm bacteria could have potential applications as antifouling agents, irrespective of irrespective of prep. Without consideration of; regardless of. irrespective of preposition despite their ecologic function. In general, the whole body extracts of the molluscs with very few exceptions showed antibacterial activity against a small proportion of biofilm bacteria. Among the inhibitory activity observed in the whole body extracts, only Nerita albicilla and Nerita oryzarum showed broad spectral activity (Table 6). The observed inhibition range of [less than or equal to]8 mm was also higher when compared with the whole body extracts of Cerithidea cingulata and Hemifusus pugilinus, which showed a range of up to 1.5 mm against nine pathogenic bacteria Pathogenic bacteria Bacteria that produce illness. Mentioned in: Gastroenteritis (Rajaganapathi 1996). The broad spectral activity observed for Nerita albicilla could be due to the previously reported occurrence of antibacterial secondary metabolites in Nerita albicilla (Sanduja et al. 1985). The bivalve whole body extracts in this study have not shown promising activity except the moderate activity observed for Pinna bicolor (72) and Pteria inquinata (74). The observation of lack of activity for Circe scripta (65) in the present study is in contrast to the observation of Jayaseeli et al. (2001) who have reported 20mm zone of inhibition for the ethanolic extract of Circe scripta against Bacillus subtilis Noun 1. Bacillus subtilis - a species of bacillus found in soil and decomposing organic matter; some strains produce antibiotics Bacillus globigii, grass bacillus, hay bacillus . Though the previous reports indicated broad spectrum activity against the human pathogenic bacteria, the extracts of bivalves in the present study seem to be ineffective or inhibit fewer biofilm bacteria. No promising antibacterial activity has been recorded for the whole body extracts and inks of cephalopods against the biofilm bacterial strains. But, broad-spectrum antibacterial activity for aqueous aqueous /aque·ous/ (a´kwe-us) 1. watery; prepared with water. 2. see under humor. a·que·ous adj. ink extract of the cephalopods Loligo duvaucelli and Sepia pharaonis against nine human pathogens has been reported (Patterson Edward & Murugan 2000). This suggests that marine biofilm bacteria are more resistant to the active compounds found in cephalopod ink. The observation of antibacterial activity in the egg masses of all the 10 gastropods in this study (Table 5) substantiates previous reports that a number of marine molluscs protect their eggs with natural products (see Kamiya et al. 1984, Matsunaga et al. 1986, Yamazaki 1993, Benkendorff et al. 2000a, Pawlik et al. 1988). The observation on Rapana rapiformis egg mass coincides with the observation of Rajaganapathi (1996) and Prem Anand et al. (1997) who have observed broad spectrum activity of the egg mass against nine pathogenic bacteria. The gelatinous egg masses of molluscs in the Aplysiidae exhibit broad spectrum of biologic activity, and several glycoproteins from the egg masses of Aplysia kurodai and A. juliana were found to have antimicrobial activity (Kamiya et al. 1984, Yamazaki et al. 1984, Yamazaki et al. 1985, Kisugi et al. 1987, Yamazaki 1993). The high antimicrobial activity of the egg masses of Chicoreus ramosus, C. virgineus, and Rapana rapiformis indicated the localization Customizing software and documentation for a particular country. It includes the translation of menus and messages into the native spoken language as well as changes in the user interface to accommodate different alphabets and culture. See internationalization and l10n. of the inhibitory substance in the egg masses. Three antimicrobial compounds namely Tyrindoleninone, Tyriverdin, and 6-bromoisatin from the egg masses of muricids have been attributed to the antimicrobial properties observed in these egg capsules (Benkendorff et al. 2001). Benkendorff et al. (2000a) observed a form of chemical ripening ripening said of meat. See curing. in the egg masses of Dicathais orbita where the toxic compound tyrindoleninone is converted into tyriverdin, a bacteristatic compound that is then converted into Tyrian Tyr´i`an a. 1. Of or pertaining to Tyre or its people. 2. Being of the color called Tyrian purple. The bright-eyed perch with fins of Tyrian dye. - Pope. purple at the time of hatching and has no antibiotic property. It is likely that the egg masses of Chicoreus ramosus, C. virgineus, and Rapana rapiformis possess the same types of bioactive compounds, because these also turn purple on exposure to sunlight. The origin of these antibacterial compounds in the egg masses has been traced to hypobranchial glands. This could be substantiated from the observation of broad-spectrum inhibitory activity of the hypobranchial gland gland, organ that manufactures chemical substances. A gland may vary from a single cell to a complex system of tubes that unite and open onto a surface through a duct. The endocrine glands, e.g. of Chicoreus virgineus against pathogenic bacteria (Rajaganapathi 1996, Prem Anand et al. 1997). Murugan et al. (1991) reported the antibacterial property of the hypobranchial gland extract of Rapana rapiformis against eight human pathogens. It is possible that the antimicrobial substances in the adult molluscs may be held in a nontoxic state and then enzymatically activated in the reproductive tract or egg masses as opined by Benkendorff (1999). This statement is substantiated by the observation of low antibacterial activity for the whole body extract of these three muricids in the present study. The activity against fewer biofilm bacteria observed in Cronia margariticola and in other gastropod egg masses of Cassidae, Conidae, Cypraeidae, Alplysidae, and Buccinidae in the present study could be influenced by their stage of development. Benkendorff et al. (2001) demonstrated that antimicrobial metabolites are likely to be more concentrated in freshly laid egg masses than in well developed or hatching egg masses. This has been demonstrated in the gelatinous egg masses of sea hare Aplysia juliana wherein the loss of antimicrobial activity was observed with embryonic development (Kamiya et al. 1988). In this study, only the egg masses with developing embryos were selected and the developmental stages have not been taken into account. Earlier studies indicate that a range of micro and macro algae and protozoans foul the egg capsules, which may have harmful or beneficial effects (Biermann et al. 1992, Benkendorff 1999, Przeslawski & Benkendorff 2005). However, the proteinaceous matrix of the leathery leath·er·y adj. Having the texture or appearance of leather: a leathery face. leath  er·i·ness n. egg
capsules appears to provide less suitable surface for fouling organisms
than gelatinous egg masses (Przeslawski & Benkendorff 2005). The
screening of empty egg cases of Chicoreus ramosus, C. virgineus, and
Rapana rapiformis did not show any inhibitory activity, indicating the
presence of active substances in only the intracapsular fluid and/or
embryos of the egg masses. The wide spectrum antibacterial activity
exhibited by the encapsulated egg masses of the three muricid gastropods
indicated the presence of potential antifouling natural products that
are worthy of further isolation and characterization.
In this study, the frequency of activity was high in methanolic extracts (Table 1-6) and acetone extract of Rapana rapiformis egg mass. The observation of lack of activity for 32.4% of the molluscan extracts may be due to limitations with the disc diffusion assay, because only hydrophilic hydrophilic /hy·dro·phil·ic/ (-fil´ik) readily absorbing moisture; hygroscopic; having strongly polar groups that readily interact with water. hy·dro·phil·ic adj. compounds tend to diffuse across the watery agar surface (Benkendorff et al. 2000b).
TABLE 1.
Proportion of biofilm bacteria inhibited by the whole body extracts of
molluscs (% inhibition against 40 biofilm bacteria).
S. No Family Molluscan Species
Gastropoda: Eogastropoda:
Patellogastropoda
1. Patellidae Cellana radiata (Born, 1778)
2. Patellidae Cellana rota (Gmelin, 1791)
Gastropoda: Orthogastropoda:
Vestigastropoda
3. Haliotidae Haliotis varia (Linne, 1758)
4. Trochidae Euchelus asper (Gmelin, 1791)
5. Trochidae Trochus radiatus (Gmelin, 1791)
6. Trochidae Umbonium vestiarium (Linne, 1758)
7. Turbinidae Turbo bruneus (Roding, 1798)
Gastropoda: Orthogastropoda:
Neritimorpha
8. Neritidae Nerita oryzarum (Recluz, 1841)
9. Neritidae Nerita albicilla (Linne, 1758)
Gastropoda: Orthogastropoda:
Caenogastropoda
10. Cypraeidae Cypraea tigiris (Linne, 1758)
11. Cypraeidae Cypraea errones (Linne, 1758)
12. Cypraeidae Cypraea onyx (Linne, 1758)
13. Cypraeidae Cypraea arabica (Linne, 1758)
14. Cypraeidae Cypraea vitellus (Linne, 1758)
15. Cypraeidae Cypraea lentiginosa (Gray, 1825)
16. Cypraeidae Cypraea caurica (Linne, 1758)
17. Cypraeidae Cypraea moneta (Linne, 1758)
18. Cypraeidae Cypraea gracilis (Gaskoin, 1848)
19. Cypraeidae Cypraea caputserpentis (Linne, 1758)
20. Cypraeidae Cypraea talpa (Linne, 1758)
21. Strombidae Tibia delicatula (Nevill, 1881)
22. Strombidae Tibia curta (Sowerby, 1842)
23. Strombidae Strombus marginatus (Linne, 1767)
24. Strombidae Lambis lambis (Linne, 1758)
25. Olividae Agaronia gibbosa (Born, 1778)
26. Muricidae Chicoreus virgineus (Roding, 1798)
27. Muricidae Chicoreus ramosus (Linne, 1758)
28. Muricidae Cronia margariticola (Broderip, 1832)
29. Muricidae Rapana rapiformis (Born, 1778)
30. Muricidae Murex tribulus (Linne, 1758)
31. Muricidae Thais biserialis. (Blainville, 1832)
32. Muricidae Thais bufo (Lamarck, 1822)
33. Muricidae Thais tissoti (Petit, 1852)
34. Volutidae Harpulina Iapponica (Linne, 1767)
35. Buccinidae Cantharus wrightae (Cernohorsky, 1974)
36. Conidae Conus araneosus (Lightfoot, 1786)
37. Conidae Conus figulinus (Linne, 1758)
38. Conidae Conus madagascariensis (Sowerby, 1858)
39. Conidae Conus textile (Linne, 1758)
40. Potamididae Cerithidea cingulata (Gmelin, 1791)
41. Littorinidae Littorarina undulata (Gray, 1839)
42. Littorinidae Littorarina scabra (Linne, 1758)
43. Fasciolariidae Fusinus forceps (Perry, 1811)
44. Cassidae Phalium glauccum (Linne, 1758)
45. Cassidae Casmaria erinacea (Linne, 1758)
46. Harpidae Harpa davidis (Roding, 1798)
47. Bursidae Bufonaria rana (Linne, 1758)
48. Naticidae Natica vitellus (Linne, 1758)
49. Naticidae Tanea tigrina (Roding, 1798)
50. Naticidae Polinices didyma (Roding, 1798)
51. Naticidae Polinices albumen (Linne, 1758)
52. Turritellidae Turitella duplicata (Linne, 1758)
53. Ranellidae Cymatium perryi (Emerson & Old, 1963)
Gastropoda: Orthogastropoda:
Heterobranchia
54. Ficidae Ficus ficus (Linne, 1758)
55. Bullidae Bulla ampulla (Linne, 1758)
56. Hydatinidae Hydatina velum (Gmelin, 1791)
57. Aplysiidae Aplysia extraordinaria (Allan, 1932)
58. Aplysiidae Dolabella auricularia (Lightfoot, 1786)
Cephalopoda: Teuthoidea
59. Loliginidae Sepioteuthis lessoniana (Lesson, 1830)
Cephalopoda: Sepioidea
60. Sepiidae Sepia pharaonis (Ehrenberg, 1831)
Bivalvia: Veneroida
61. Donacidae Donax faba (Gmelin, 1791)
62. Donacidae Donax cuneatus (Linne, 1758)
63. Veneridae Gafrarium tumidum (Roding, 1798)
64. Veneridae Gafrarium pectinatum (Linne, 1758)
65. Veneridae Circe scripta (Linne, 1758)
66. Veneridae Dosinia modesta (Sowerby)
67. Veneridae Marcia opima (Gmelin, 1791)
68. Veneridae Meretrix casta (Gmelin, 1791)
69. Mesodesmatidae Atactodea striata (Gmelin, 1791)
70. Cardiidae Vasticardium pectiniforme (Born, 1778)
71. Carditidae Cardita bicolor (Linne, 1758)
Bivalvia: Pterioida
72. Pinnidae Pinna bicolor (Gmelin, 1791)
73. Pteriidae Pteria castanea (Reeve, 1858)
74. Pteriidae Pteria inquinata (Reeve, 1857)
Bivalvia: Mytiloida
75. Mytilidae Modiolus metcalfei (Hanley, 1843)
Bivalvia: Ostreoida
76. Spondylidae Spondylus layardi (Reeve, 1856)
77. Placunidae Placuna placenta (Lamarck, 1819)
Type of the Extract
S. No M MW MD A
1. -- -- 20 --
2. -- -- 23 --
3. -- -- -- --
4. -- 30 -- --
5. -- -- -- --
6. -- -- -- --
7. 73 -- -- --
8. -- 95 -- --
9. -- 93 -- --
10. -- -- 48 --
11. -- -- 40 --
12. -- -- 28 --
13. -- -- 25 --
14. -- -- 30 --
15. -- -- -- --
16. -- -- -- --
17. -- -- -- --
18. -- -- 45 --
19. -- 38 -- --
20. -- -- -- --
21. -- 40 -- --
22. -- 55 -- --
23. -- 70 -- --
24. -- -- -- 55
25. -- 40 -- --
26. 50 -- -- --
27. 58 -- -- --
28. -- -- 63 --
29. -- -- -- 28
30. -- -- -- 60
31. 35 -- -- --
32. 25 -- -- --
33. 18 -- -- --
34. 13 -- -- --
35. -- -- -- --
36. 33 -- -- --
37. 30 -- -- --
38. 15 -- -- --
39. 33 -- -- --
40. -- -- -- --
41. -- -- 28 --
42. -- -- 25 --
43. -- -- 30 --
44. -- -- -- --
45. 20 -- -- --
46. -- -- -- --
47. 63 -- -- --
48. -- -- -- --
49. -- -- -- --
50. -- -- -- --
51. -- -- -- --
52. 33 -- -- --
53. -- -- -- --
54. 45 -- -- --
55. 40 -- -- --
56. -- -- -- --
57. 28 -- -- --
58. 38 -- -- --
59. 40 -- -- --
60. 30 -- -- --
61. -- 25 -- --
62. -- 28 -- --
63. 33 -- -- --
64. 20 -- -- --
65. -- -- -- --
66. -- 25 -- --
67. -- 38 -- --
68. -- 30 -- --
69. -- -- -- --
70. 8 -- -- --
71. -- -- -- --
72. -- -- 55 --
73. 30 -- -- --
74. 45 -- -- --
75. -- -- -- --
76. -- -- -- --
77. -- 10 -- --
M, Methanol; MW, Methanol: Water; MD, Methanol: Dichloromethane; A,
Acetone.
TABLE 2.
Proportion of biofilm bacteria inhibited by the molluscan
digestive gland extracts (% inhibition against 40 biofilm bacteria).
S. No Family Molluscan Species
Gastropoda: Orthogastropoda:
Vestigastropoda
78. Haliotidae Haliotis varia (Linne, 1758)
Gastropoda: Orthogastropoda:
Caenogastropoda
79. Fasciolariidae Pleuroploca trapezium (Linne, 1758)
80. Tonnidae Tonna dolium (Linne, 1758)
81. Volutidae Melo melo (Lightfoot, 1786)
82. Melongenidae Hemifusus pugilinus (Born, 1778)
83. Buccinidae Babylonia spirata (Linne, 1758)
84. Buccinidae Babylonia zeylanica (Bruguiere, 1789)
85. Muricidae Chicoreus virgineus (Roding, 1798)
86. Muricidae Chicoreus ramosus (Linne, 1758)
87. Strombidae Lambis lambis (Linne, 1758)
Type of the Extract
S. No M MW MD A
78. 23 -- -- --
79. -- -- -- --
80. -- -- -- --
81. 28 -- -- --
82. -- -- -- --
83. -- -- -- --
84. -- -- -- --
85. -- -- -- --
86. -- -- -- --
87. -- -- -- --
M, Methanol; MW, Methanol: Water; MD, Methanol: Dichloromethane; A,
Acetone.
TABLE 3.
Proportion of biofilm bacteria inhibited by the molluscan ink extracts
(% inhibition against 40 biofilm bacteria).
S. No Family Molluscan Species
Gastropoda: Orthogastropoda:
Heterobranchia
88. Aplysiidae Aplysia extraordinaria (Allan, 1932)
89. Aplysiidae Dolabella auricularia (Lightfoot, 1786)
Cephalopoda: Teuthoidea
90. Loliginidae Sepioteuthius lessoniana (Lesson, 1830)
Cephalopoda: Sepioidea
91. Sepiidae Sepia pharaonis (Ehrenberg, 1831)
Type of the Extract
S. No M MW MD A
88. 30 -- -- --
89. 43 -- -- --
90. -- -- -- --
91. -- -- -- --
M, Methanol; MW, Methanol: Water; MD, Methanol: Dichloromethane; A,
Acetone.
TABLE 4.
Proportion of biofilm bacteria inhibited by the molluscan opercula
extracts (% inhibition against 40 biofilm bacteria).
S. No Family Molluscan Species
Gastropoda: Orthogastropoda: Neritimorpha
92. Neritidae Nerita oryzarum (Recluz, 1841)
Gastropoda: Orthogastropoda: Caenogastropoda
93. Strombidae Lambis lambis (Linne, 1758)
94. Buccinidae Babylonia spirata (Linne, 1758)
95. Muricidae Chicoreus virgineus (Roding, 1798)
Type of the Extract
S. No M MW MD A
92. -- -- -- --
93. 15 -- -- --
94. -- -- -- --
95. 35 -- -- --
M, Methanol; MW, Methanol: Water; MD, Methanol: Dichloromethane; A,
Acetone.
TABLE 5.
Proportion of biofilm bacteria inhibited by the encapsulated molluscan
egg mass extracts (% inhibition against 40 biofilm bacteria).
S. No Family Molluscan Species
Gastropoda: Orthogastropoda:
Caenogastropoda
96. Muricidae Chicoreus virgineus (Roding, 1798)
97. Muricidae Chicoreus ramosus (Linne, 1758)
98. Muricidae Cronia margariticola (Broderip, 1832)
99. Muricidae Rapana rapiformis (Born, 1778)
100. Cassidae Casmaria erinacea (Linne, 1758)
101. Conidae Conus madagascariensis (Sowerby, 1858)
102. Cypraeidae Cypraea errones (Linne, 1758)
103. Buccinidae Babylonia spirata (Linne, 1758)
Gastropoda: Orthogastropoda:
Heterobranchia
104. Aplysiidae Aplysia extraordinaria (Allan, 1932)
105. Aplysiidae Dolabella auricularia (Lightfoot, 1786)
Type of the Extract
S. No M MW MD A
96. 100 -- -- --
97. 100 -- -- --
98. 70 -- -- --
99. -- -- -- 100
100. 55 -- -- --
101. 53 -- -- --
102. 35 -- -- --
103. 35 -- -- --
104. 40 -- -- --
105. 28 -- -- --
M, Michael; MW, Methanol; Water; MD, Methanol; Dichloromethane; A,
Acetone.
TABLE 6.
Zone of inhibition (mm) of five molluscan crude extracts against 40
biofilm bacteria.
Inhibition Zone (mm)
Chicoreus
Chicoreus Rapana ramosus
virgineus rapiformis (Egg Mass)
Biofilm Bacteria (Egg Mass)(M) (Egg Mass)(A) (M)
Micrococcus sp. 1 7 5 3
Micrococcus sp. 2 5 4 3
Micrococcus sp. 3 4 2 1
Micrococcus sp. 4 3 2 1
Vibrio sp. 1 8 6 4
Vibrio sp. 2 10 7 5
Vibrio sp. 3 5 4 2
Vibrio sp. 4 3 2 1
Vibrio sp. 5 2 2 1
Pseudomonas sp. 1 6 4 3
Pseudomonas sp. 2 5 4 3
Pseudomonas sp. 3 6 3 3
Pseudomonas sp. 4 2 1 2
Alcaligenes sp. 1 3 1 1
Alcaligenes sp. 2 2 2 2
Alcaligenes sp. 3 6 5 3
Flavobacterium sp. 1 4 3 3
Flavobacterium sp. 2 3 1 1
Flavobacterium sp. 3 6 5 1
Flavobacterium sp. 4 7 4 1
Aeromonas sp. 1 4 3 2
Aeromonas sp. 2 8 3 2
Aeromonas sp. 3 7 5 2
Aeromonas sp. 4 3 3 3
Bacillus sp. 1 6 1 1
Bacillus sp. 2 3 2 3
Bacillus sp. 3 2 1 1
Bacillus sp. 4 4 1 1
Enterobacter sp. 1 4 3 4
Enterobacter sp. 2 5 1 3
Enterobacter sp. 3 8 4 5
Enterobacter sp. 4 2 2 1
Alteromonas sp. 1 5 2 3
Alteromonas sp. 2 7 5 4
Alteromonas sp. 3 4 3 3
Alteromonas sp. 4 6 4 4
Cornebacterium sp. 1 2 2 1
Cornebacterium sp. 2 1 1 1
Cornebacterium sp. 3 2 1 1
Cornebacterium sp. 4 4 2 1
Inhibition Zone (mm)
Nerita
Nerita oryzarum
albicilla (Whole Body)
Biorilm Bacteria (Whole Body)(MW) (MW)
Micrococcus sp. 1 1 2
Micrococcus sp. 2 2 4
Micrococcus sp. 3 4 1
Micrococcus sp. 4 1 1
Vibrio sp. 1 1 Trace
Vibrio sp. 2 1 1
Vibrio sp. 3 2 2
Vibrio sp. 4 2 2
Vibrio sp. 5 2 1
Pseudomonas sp. 1 1 1
Pseudomonas sp. 2 1 3
Pseudomonas sp. 3 Nil 1
Pseudomonas sp. 4 Trace Nil
Alcaligenes sp. 1 Trace 3
Alcaligenes sp. 2 2 3
Alcaligenes sp. 3 1 2
Flavobacterium sp. 1 Trace 1
Flavobacterium sp. 2 3 Trace
Flavobacterium sp. 3 3 4
Flavobacterium sp. 4 Trace Trace
Aeromonas sp. 1 1 8
Aeromonas sp. 2 4 3
Aeromonas sp. 3 4 3
Aeromonas sp. 4 2 2
Bacillus sp. 1 Trace Trace
Bacillus sp. 2 Nil Trace
Bacillus sp. 3 Nil Trace
Bacillus sp. 4 Trace 1
Enterobacter sp. 1 2 1
Enterobacter sp. 2 2 1
Enterobacter sp. 3 1 1
Enterobacter sp. 4 1 1
Alteromonas sp. 1 3 2
Alteromonas sp. 2 1 Nil
Alteromonas sp. 3 1 Trace
Alteromonas sp. 4 2 2
Cornebacterium sp. 1 2 3
Cornebacterium sp. 2 1 1
Cornebacterium sp. 3 3 1
Cornebacterium sp. 4 1 1
M, Methanol extracts; A, Acetone extracts; MW, Methanol: Water
extracts.
ACKNOWLEDGMENTS The first author thanks DANIDA DANIDA Danish International Development Agency and Prof. Jorgen Hylleberg of Aarhus University, Denmark, for the fellowship through Tropical Marine Mollusc Programme (TMMP TMMP Tokyo Mew Mew Power (anime) TMMP Technical Manual Management Program/Plan TMMP Ttissue Matrix Metalloproteinase ). The authors thank Dr. Alan Kohn, University of Washington, USA and Mr. Peter Middelfart of Australian Museum, Australia, for help in identification of specimens. The authors also gratefully acknowledge the constructive review of the manuscript by Dr. Kirsten Benkendorff of School of Biological Sciences, Flinders University The university has established a reputation as a leading research institution with a devotion to innovation. It is a member of Innovative Research Universities Australia and ranks among the leading universities in Australia. , Australia. LITERATURE CITED Barroso, C. M., M. A. Reis-Henriques, M. S. Ferreira & M. H. Moreira. 2002. The effectiveness of some compounds derived from antifouling paints Noun 1. antifouling paint - a paint used to protect against the accumulation of barnacles etc. on underwater surfaces paint, pigment - a substance used as a coating to protect or decorate a surface (especially a mixture of pigment suspended in a liquid); dries to in promoting imposex in Nassarius reticulates. J. Mar. Biol. Assoc. UK. 82:249-255. Biermann, C., G. Schinner & R. Strathmann. 1992. Influence of solar radiation solar radiation, n the emission and diffusion of actinic rays from the sun. Overexposure may result in sunburn, keratosis, skin cancer, or lesions associated with photosensitivity. , microalgal fouling, and current on deposition site and survival of embryos of a dorid nudibranch nudibranch: see sea slug. nudibranch or sea slug Any marine gastropod in the order Nudibranchia. Most nudibranchs lack a shell, mantle cavity (see mollusk), and gills, and breathe through the body surface. gastropod. Mar. Ecol. Progr. Ser. 86:205-215. Becerro, M. A., N. I. Lopez, X. Turon & M. J. Uniz. 1994. Antimicrobial activity and surface bacterial film in marine sponges. J. Exp. Mar. Biol. Ecol. 179:195-205. Belfroid, A. C., M. Purperhart & F. Ariese. 2000. Organotin levels in seafood. Mar. Pollut. Bull. 40(3):226-232. Benkendorff, K. 1999. Bioactive molluscan resources and their conservation: Ph.D. Thesis, University of Wollongong History The University of Wollongong was founded in 1951 when a Division of the then New South Wales University of Technology (re-named the University of New South Wales in 1958) was established in Wollongong. , Australia. Available at: http://www.library.uow.edu/adt-NWU/public/adt-NWU20011204. 154039/index.html. Benkendorff, K., J. B. Bremner & A. R. Davis. 2000a. Tyrian purple precursors in the egg masses of the Australian muricid, Dicathais orbita: a possible defensive role. J. Chem. Ecol. 26:1037-1050. Benkendorff, K., A. R. Davis & J. B. Bremner. 2000b. Rapid screening methods for detecting antimicrobial activity in the egg masses of marine molluscs. J. Med. & Appl. Malacol. 10:211-223. Benkendorff, K., A. Davis & J. B. Bremner. 2001. Chemical defense in the egg masses of benthic ben·thos n. 1. The collection of organisms living on or in sea or lake bottoms. 2. The bottom of a sea or lake. [Greek. invertebrates: An assessment of antibacterial activity in 39 Molluscs and 4 Polychaetes. J. Invertebr. Pathol. 78: 109-118. Bettin, C., J. Oehlmann & E. Stroben. 1996. TBT-induced imposex in marine neogastropods is mediated me·di·ate v. me·di·at·ed, me·di·at·ing, me·di·ates v.tr. 1. To resolve or settle (differences) by working with all the conflicting parties: by an increasing androgen androgen (ăn`drəjən): see testosterone. androgen Any of a group of hormones that mainly influence the development of the male reproductive system. level. Helgolander Meeresunters 50:299-317. Cimino, G. & G. Sodano. 1993. Biosynthesis Biosynthesis The synthesis of more complex molecules from simpler ones in cells by a series of reactions mediated by enzymes. The overall economy and survival of the cell is governed by the interplay between the energy gained from the breakdown of compounds of secondary metabolites in marine molluscs. Top. Curr. Chem. 167:77-115. Clare, A. 1996. Marine natural product antifoulants: status and potential. Biofouling 9:211-229. De Nys, R. & P. D. Steinburg. 2002. Linking marine biology marine biology, study of ocean plants and animals and their ecological relationships. Marine organisms may be classified (according to their mode of life) as nektonic, planktonic, or benthic. Nektonic animals are those that swim and migrate freely, e.g. and biotechnology. Current Opinion in Biotechnology 13:244-248. Devi, P., W. Solimabi, L. D'Souza, S. Sonal, S. Y. Kamar & S. Y. S. Singbal. 1997. Screening of some marine plants for activity against marine fouling bacteria. Bot. Mar. 40:87-91. Faulkner, D. J. 1988. Feeding deterrent in Molluscs. in: D. G. Fautin, editor. Biomedical bi·o·med·i·cal adj. 1. Of or relating to biomedicine. 2. Of, relating to, or involving biological, medical, and physical sciences. importance of marine organisms. California Academy of Sciences The California Academy of Sciences is one of the ten largest natural history museums in the world, and one of the oldest in the United States of America. It is located in Golden Gate Park in San Francisco. . pp. 29-36. Fletcher, M. & K. C. Marshall. 1982. Are solid surfaces of ecological significance to aquatic bacteria? Adv. Microb. Ecol. 6:199-236. Fusetani, N. 2004. Biofouling and antifouling. Nat. Prod. Rep. 21:94-104. Holt, J. G., N. R. Krieg, P. H. Sneath, J. T. Staley & S. T. Williams. 1994. International edition: Bergey's manual of determinative bacteriology bacteriology Study of bacteria. Modern understanding of bacterial forms dates from Ferdinand Cohn's classifications. Other researchers, such as Louis Pasteur, established the connection between bacteria and fermentation and disease. . 9th ed. Maryland: Williams & Wilkine. Hellio, C., J.-P. Marechal, B. Veron, G. Bremer, A. S. Clare & Y. L. Gal. 2004. Seasonal variation of antifouling activities of marine algae from the Brittany Coast (France). Mar. Biotechnol. 6:67-82. Hylleberg, J. & R. N. Kilburn. 2002. Annotated inventory of molluscs from the Gulf of Mannar The Gulf of Mannar is a large shallow bay that is an arm of the Indian Ocean. It lies between the southeasternern tip of India and the west coast of Sri Lanka with widths between 160 and 200 km (100 to 125 mi). and vicinity. In: J. Hylleberg & A. Nateewathana, editors. Zoogeography zoogeography defining the location and numbers of animal populations, and their variability with time. and inventory of marine molluscs encountered in Southern India. PMBC PMBC Pasadena Mountain Bike Club (California) PMBC Physical Model Based Control PMBC Program Managers Briefing Course , Thailand. Phuket. Mar. Biol. cent. spec. publ. 29. pp. 19-70. Jayaseeli, A. A., T. Prem Anand & A. Murugan. 2001. Antibacterial activity of 4 bivalves from Gulf of Mannar. Phuket. Mar. Biol. Cent. Spec. Publ. 25(1):215-217. Kamiya, H., K. Muramoto & K. Ogata. 1984. Antibacterial activity in the egg mass of a sea hare. Experimentia 40:947. Kamiya, H., K. Muramoto, R. Goto & M. Yamazaki. 1988. Characterization of the antibacterial and antineoplastic antineoplastic /an·ti·neo·plas·tic/ (-ne?o-plas´tik) 1. inhibiting or preventing development of neoplasms; checking maturation and proliferation of malignant cells. 2. an agent that so acts. glycoproteins in a sea hare Aplysia juliana. Nippon Suisan Gakkaishi 54:773-777. Kisugi, J., H. Kamiya & M. Yamazaki. 1987. Purification and Characterization of Aplysianin E, an antitumour factor from sea hare eggs. Cancer Res. 47:5649-5653. Koh. 1997. Do scleratinian corals engage in chemical warfare chemical warfare, employment in war of incendiaries, poison gases, and other chemical substances. Ancient armies attacking or defending fortified cities threw burning oil and fireballs. A primitive type of flamethrower was employed as early as the 5th cent. B.C. against microbes? J. Chem. Ecol. 23:379-398. Lord, F. 1986. Are the contents of egg capsules of the marine gastropod Nucella lapillus (L) axenic. Am. Malac. Bull. 4:201-203. Matsunaga, S., N. Fusetani & K. Hashimoto. 1986. Kabiramide C, a novel antilungal macrolide from nudibranch egg masses. J. Am. Chem. Soc. 108:847-849. Mitta, G., F. Vandenbulcke, F. Hubert, M. Salzet & P. Roche. 2000. Involvement of mytilins in mussel antimicrobial defense. J. Biol. Chem. 275:12954-12962. Murugan, A. & M. Santhana Ramasamy. 2003. Biofouling deterrent natural product from the ascidian ascidian: see Chordata; tunicate. Distaplia nathensis. Indian J. Mar. Sci. 32:162-164. Murugan, A., J. K. Patterson Edward & K. Ayyankannu. 1991. Ecological, biochemical and antibacterial data from India. Phuket Mar. Biol. Cent. Spec. Publ. No. 9:108-110. Newbold, R. W., P. R. Jensen, W. Fenical & J. R. Pawlik. 1999. Antibacterial activity of Caribbean sponge extracts. Aquat. Microb. Ecol. 19: 279-284. Patterson Edward, J. K. & A. Murugan. 2000. Screening of cephalopods for bioactivity. Phuket Mar. Biol. Cent. Spec. Publ. No. 21 (1):253-256. Pawlik. J. R., 1993. Marine invertebrate invertebrate (ĭn'vûr`təbrət, –brāt'), any animal lacking a backbone. The invertebrates include the tunicates and lancelets of phylum Chordata, as well as all animal phyla other than Chordata. chemical defenses. Chem. Rev 93:1911-1922. Pawlik, J. R., M. R. Kernan, T. F. Molinski & M. K. Harper. 1988. Defensive chemicals of the Spanish Dancer The Spanish dancer, Hexabranchus sanguineus ("Bloody Six-Gills"), is a nudibranch gastropod of the family Hexabranchidae. It is a large, strong swimming species found throughout the tropical Indo-Pacific and Red Sea. nudibranch Hexabranchus sanguineus, and its egg ribbons, Macrolids derived from a sponge diet. J. Exp. Mar. Biol. Ecol. 119:99-109. Prem Anand, T., J. Rajaganapathi & J. K. Patterson Edward. 1997. Antibacterial activity of Marine Molluscs from Portonovo region. Indian J. Mar. Sci. 26:206-208. Przeslawski, R. & K. Benkendorff. 2005. The role of surface fouling in the development of encapsulated gastropod embryos. J. Mollusc. Stud. 71: 75-83. Rajaganapathi, J. 1996. Studies on antibacterial activity of five marine molluscs. M.Sc. thesis. Annamalai University Annamalai University is a state university in Annamalai Nagar, Tamil Nadu, India. History The Annamalai University owes its existence to the vision, wisdom, benefaction and philanthropy of Dr. Rajah Sri Annamalai Chettiar. , Parangipettai. India, pp. 43. Riguera, R. 1997. Isolating bioactive compound from marine organisms. J. Mar. Biotechnol. 5:187-193. Rinehart, K. L., P. D. Shaw, L. S. Shield, J. B. Gloer, G. C. Harbour, M. E. S. Koker, D. Samain, R. E. Schwartz, A. A. Tymiak, E. G. Swynenberg, D. A. String Fellow, J. J. Vavva, J. H. Coats, G. E. Zurenko, S. L. Kuentzel, L. H. Li, G. J. Bakus, R. C. Brasca, L. L. Craft, D. N. Young & J. L. Connot. 1981. Marine natural products as a source of antiviral antiviral /an·ti·vi·ral/ (-vi´ral) destroying viruses or suppressing their replication, or an agent that so acts. an·ti·vi·ral adj. , antimicrobial and antineoplastic agents antineoplastic agent, n a drug that prevents the development, maturation, or spread of neoplastic cells. . Pure Appl. Chem. 53:795-817. Rogers, C., R. De Nys, T. Charlton & P. Steinberg. 2000. Dynamics of algal algal pertaining to or caused by algae. algal infection is very rare but systemic and udder infections are recorded. See protothecosis. algal mastitis the algae Prototheca trispora and P. secondary metabolites in two species of sea hare. J. Chem. Ecol. 26:721-744. Sanduja, R., A. J. Weinheimer, K. L. Euler & M. Alam. 1985. Unusual occurrence of faloplumierin, an antibacterial agent in the marine mollusc Nerita albicilla. J. Natl. Prod. 48:335-336. Santhana Ramasamy, M. & A. Murugan. 2003. Chemical defense in ascidians Eudistoma viride and Didemnum psammathodes in Tuticorin, Southeast coast of India: Bacterial epibiosis and fouling deterrent activity. Indian J. Mar. Sci. 32(4):337-339. Satyamurti, S. J. 1952 a & b. The mollusca of Krusadi Island (in the Gulf of Mannar). I. Amphineura and Gastropoda; II. Scaphapoda, Pelecypoda and Cephalopoda. pp. 466. Scheltema, R. S. 1974. Biological interactions determining larval larval 1. pertaining to larvae. 2. larvate. larval migrans see cutaneous and visceral larva migrans. settlement of marine invertebrates. Thalassia Jugosl. 10:263-269. Slattery, M., J. B. McClintock & J. N. Heine. 1995. Chemical defenses in Antarctic soft corals: Evidence for antifouling compounds. J. Exp. Mar. Biol. Ecol. 190:61-77. Steinberg, P. D., R. De Nys & S. Kjelleberg. 2002. Chemical cues for surface colonization. J. Chem. Ecol. 28(10):1935-1951. Tang, R. J. & J. J. Cooney. 1998. Effects of marine paints on microbial biofilm development at three materials. J. Ind. Microbiol. Biotechnol. 20:275-280. Wahl, M. 1995. Bacterial epibiosis on Bahamian and Pacific ascidians. J. Exp. Mar. Biol. Ecol. 191:239-255. Wahl, M., K. Kroger & M. Lenz. 1998. Nontoxic protections against epibiosis. Biofouling 12(13):205-226. Wright, A. E. 1998. Isolation of marine natural products. In: R. P. J. Cannell, editor. Methods in biotechnology, vol. 4. Natural product isolation. New Jersey: Humana press Inc. pp. 365-408. Yamazaki, M. 1993. Antitumor an·ti·tu·mor also an·ti·tu·mor·al adj. Counteracting or preventing the formation of malignant tumors; anticancer. Adj. 1. and antimicrobial glycoprotens from seahares. Comp. Biochem. Physiol. 105C:141-146. Yamazaki, M., J. Kisugi, H. Kimura & D. Mizuno. 1985. Purification of antineoplastic factor from eggs of a seahare. FEBS FEBS Federation of European Biochemical Societies Lett. 185:295-298. Yamazaki, M., J. Kisugi, M. Ikenami, H. Kamiya & D. Mizuno. 1984. Cytolytic cytolytic pertaining to or emanating from cytolysis. cytolytic reactivity type II hypersensitivity. factor in eggs of the seahare Aplysia kutodai. Gann. 75:269-274. M. SANTHANA RAMASAMY AND A. MURUGAN * Suganthi Devadason Marine Research Institute, 44-Beach Road, Tuticorin-628 001, Tamil Nadu Tamil Nadu (tăm`əl nä`d  ), formerly Madras (mədrăs`, mədräs`), state (2001 provisional pop. , India
* Corresponding author. E-mail: muruganrsa@sanchamet.in |
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