Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.     

Antibacterial,antiviral and antioxidant activities of aerial part extracts of Peganum harmala L. grown in Tunisia

Ethyl acetate, chloroform, butanol, and methanol extracts from the aerial part of Peganum harmala were tested for antibacterial, antioxidant, and antiviral activities. The antibacterial activity was evaluated by the determination of minimum inhibitory concentration (MIC) using the solid medium technique. Oxacillin, amoxicillin, ticarcillin, cefotaxim, and amphotericin were used as control agents. The antiviral activity was evaluated against human cytomegalovirus (HCMV) strain AD-169 (ATCC Ref. VR 538) and Coxsackie B virus type 3 (CoxB-3) using a cytopathic effect (CPE) reduction assay. The antioxidant activity was evaluated using two tests: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical scavenging and ammonium thiocyanate methods. Among tested extracts; the chloroform extract displayed a higher antibacterial activity against Gram-positive than Gram-negative bacteria. The butanol extract demonstrated the highest antioxidant activity. The methanol extract showed significant antiviral activity against CoxB-3 virus. The chloroform extract may be an important source of bactericidal compounds against Gram-positive bacteria.     

Stress-induced rapid release of antibacterials by scleractinian corals

Mechanical stress on the coral Pocillopora damicornis caused the release of material that killed the coral pathogen Vibrio coralliilyticus. The bactericidal material was released into the surrounding seawater rapidly, reaching a maximum killing activity within 1 min of the stress. The coral antibacterial activity, referred to as CAA, was retained following filter sterilization and storage at –20°C. Exposure of V. coralliilyticus to CAA for 30 s, 1 min and 6 min resulted in the death of 82%, 89% and 99% of the bacteria, respectively. Release of CAA following mechanical stress was also observed with four other coral species tested. P. damicornis CAA was bactericidal to a wide variety of Gram-negative and Gram-positive bacteria. This is the first report that hard corals rapidly release fast-acting bactericidal material following mechanical stress. The release of CAA was demonstrated with both aquarium corals and corals taken directly from the sea. It is suggested that CAA is part of hard corals host defense system against infection, the natural stimulation for release of CAAs being the bite of a predator. Previous failures to detect antibacterial activity in hard corals can be attributed to a lack of understanding of the sensitive mechanism by which they are released.Communicated by M. Kühl, Helsingør     

Antibacterial activity of the marine blue-green alga Trichodesmium erythraeum in the gastro-intestinal contents of the sea gull Laurus brunicephalus

The gastro-intestinal contents of the sea gull Laurus brunicephalus Jerdon were found to possess antibacterial activity. This activity could be traced to heavy accumulations of the marine blue-green alga Trichodesmium erythraeum in the gut. During 1969, a bloom of T. erythraeum began in Porto Novo waters about the middle of February, and attained a peak during the second week of March. It has been reported previously by the author that antibacterial properties are exhibited by T. erythraeum maintained in laboratory cultures, as well as in water samples collected from a red tide area. It was also reported (Ramamurthy, 1970) that the gut contents in 2 pelagic fishes, Hilsa kanagurta and Rastrelliger kanagurta, collected during the same red tide bloom period, possessed antibacterial properties. During this period large numbers of these fishes were consumed by sea gulls L. brunicephalus. In view of this finding, experimental procedures were adopted to determine whether extracts of T. erythraeum occurring in the gut of the sea gulls might exhibit antibacterial activity. It was found that T. erythraeum collected from the gut of L. brunicephalus could inhibit both gram positive and gram negative bacteria. Gastro-intestinal extracts from L. brunicephalus collected during the non-bloom period of T. erythraeum showed heavy microbial growth of bacteria and fungi. Evidently, antibacterial or sterile conditions prevail in the gut of these tropical sea gulls in a manner similar to that observed in Polar penguins by Sieburth (1959, 1961).     

Bacterial colonization of the phyllosphere of nineteen plant species and antimicrobial activity of their leaf secondary metabolites against leaf associated bacteria

Summary. The scope of this work was to examine whether leaf constitutive secondary metabolites play a role in determining bacterial colonization of the phyllosphere. To this aim, we surveyed nineteen native or cultivated plant species that share a common bacterial pool in a North Mediterranean area, and estimated the size of total and ice nucleation active (INA) bacterial populations on their leaves. Large differences in the colonization of their phyllosphere were found; the population size of epiphytic bacteria ranged from 7.5 × 10² to 1 × 10⁶ CFU/g fresh weight, in eucalypt and celery, respectively. Species native in Mediterranean-type climate areas, particularly those belonging to the group of aromatic plants, are characterized by scarce presence of INA bacteria. The antibacterial activity of essential oils, surface phenolics and leaf tissue extracts was also estimated against the INA strains P. syringae and E. herbicola, isolated from two of these plant species. E. herbicola proved more sensitive than P. syringae. Of the species examined, oregano [Origanum vulgare L. subsp. hirtum (Link.) Ietswaart], an aromatic plant, had the highest antimicrobial activity, whereas six species showed no activity at all. Further experiments were performed with oregano and bean (Phaseolus vulgaris L.) that represent two extremes in their secondary metabolite content. Both plants were inoculated with P. syringae. By the end of incubation, the bacterial population on bean plants was about 100 times higher than that on oregano leaves. Scanning electron micrographs showed that bacterial growth on oregano leaves was confined to sites away from glandular hairs. Results from the bacterial colonization survey together with those from the toxicity tests showed that all species rich in antibacterial secondary metabolites harbored low leaf bacterial populations. These results provide substantial evidence that leaf secondary metabolites function as constitutive defense chemicals against microbial invasions. However, the fact that species with non- or moderately active leaf secondary metabolites are not always highly colonized suggests mediation of other unknown factors, the contribution of which requires further investigation.     

The influence of storage on antibacterial activity of sea water. I. Experiments with sea water stored at 18°C

Three sea-water samples of about 25 l each, stored at 18°C in the dark, were examined for antibacterial activity and bacterial content. Serratia marinorubra mainly was used as test bacterium. During storage for 197 to 454 days, bactericidal capacity varied considerably. In raw sea water, extreme log (N ₃–N ₀) values (N: number of colony-forming test bacteria/ml after 0 and 3 days of any test) were higher than-1 and lower than-4. Antibacterial activity of filtersterilized and autoclaved, stored, sea water likewise varied, but to a lesser degree. The number of colony-forming marine bacteria fluctuated about 3 orders of magnitude. Variations in bacterial content and in bactericidal action of stored sea water were positively correlated during the first months of storage; later, this correlation became negative. From these results it is concluded that variations in nutrient concentration — due to successive periods of growth and autolysis of marine bacteria — were the primary reason for the changes observed in antibacterial activity of stored sea-water samples.     

Seasonal changes in antibacterial activity of North Sea water

During 1969 and 1970, the antibacterial activity of North Sea water was investigated at Helgoland (southern North Sea). Tests were performed with resting cells of Escherichia coli, Staphylococous aureus and Serratia marinorubra, inoculated in freshly sampled sea water used raw or after filter sterilization. Simultaneously, growth of indigenous bacterial populations which occurred in the presence, as well as in the absence (only 1970), of test bacteria was controlled. Seasonal changes in inactivation of test cells and multiplication of marine bacteria were correlated with the life cycles of several diatom species. Breakdown of phytoplankton flowerings produced the most pronounced influence on antibacterial activity of sea water. Sometimes, enhancement of kill rate could be established during growth periods of various algal species, too. From the results obtained, it is concluded that the inactivation capacity of sea water depends, to a considerable degree, on the availability of nutritive organic matter, the concentration of which changes according to phytoplankton development. Strain-specific differences in resistance to inimical action of sea water are, in part, correlated with the ability of test cells to use, and to compete with, marine bacteria for nutrients from the sea. A somewhat modified interpretation of antibacterial activity of sea water is presented.     

Microbiological evaluation of 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one with a comparison to antibacterial activity of standard drugs

Nucleophilic substitution of 4-chloro-3-nitro-2-oxo-2H-chromen-2-one 3 by 6-chloro-7H-purin-2-ylamine (a), yielded the corresponding 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one 4. The chemical structure of synthesized compound was characterized using IR and NMR spectra, and elemental analyses. The purified synthesized compound 4 was tested at concentrations 1, 3, 5 mg/ml for its antibacterial activity against three bacterial cultures: Staphylococcus aureus, Escherichia coli, and Bacillus cereus. The antibacterial activity of synthesized compound was compared to antibacterial activity of standard antibiotics cephalexine and streptomycin.     

Antibacterial and antimutagenic activitiy of extracts and essential oil from (Tunisian) Pistacia lentiscus

Aqueous and flavonoid-enriched extract as well as essential oil (EO) obtained from leaves of Pistacia lentiscus were assessed for antibacterial and antimutagenic activities. Antibacterial activity of different extracts and EO were evaluated against six bacterial strains. A marked inhibitory effect was observed against Salmonella typhimurium, whereas lower activity was observed against Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella enteritidis. EO showed significant inhibitory effects against Salmonella typhimurium, Salmonella enteritidis and Staphylococcus aureus. The antimutagenic activity of the different extracts against Aflatoxin B₁ (AFB₁) and sodium azide was demonstrated with the Salmonella typhymurium assay. The number of revertants per plate decreased significantly when the plant extracts were added to the assay system using Salmonella typhimurium TA100, TA98 and TA1535.     

纳米银复合材料与抗生素的联合抗菌性能及相关机制研究

抗生素的滥用导致细菌耐药问题日益严重,人类迫切需要开发出新的抗菌药物以减少细菌耐药问题。基于纳米银制备而成的纳米银复合材料在兼顾纳米银抗菌性能的同时不仅能够克服单一纳米银释放速度快、不稳定等缺点,还能缓解细菌耐药的问题,因此被认为是一类具有广泛应用前景的新型抗菌剂。已有研究表明,单一纳米银与某些抗生素的联合使用可以达到协同抗菌效果,但目前尚缺乏对纳米银复合材料与抗生素的联合抗菌性能及机制的研究。本文首先制备出3种不同结构的纳米银复合材料,包括二氧化硅-聚多巴胺-纳米银复合材料(SiO_2-PD-AgNPs)、纳米银@二氧化硅复合材料(AgNPs@SiO_2)和纳米银@二氧化硅-聚多巴胺-纳米银复合材料(AgNPs@SiO_2-PD-AgNPs)。随后测定了纳米银复合材料对大肠杆菌(Escherichia coli, E. coli)和枯草芽孢杆菌(Bacillus subtilis, B. subtilis)的单一毒性效应。结果显示,AgNPs@SiO_2-PD-AgNPs复合材料对2种菌的单一毒性均大于其余2种纳米银复合材料。因此,笔者以AgNPs@SiO_2-PD-AgNPs作为代表,测定了纳米银复合材料与硫酸卡那霉素(kanamycin sulfate, KS)/盐酸土霉素(oxytetracycline hydrochloride, OH)的二元联合抗菌性能,发现AgNPs@SiO_2-PD-AgNPs与KS联合可以对E. coli产生协同效应。协同效应产生的主要原因可能是:AgNPs@SiO_2-PD-AgNPs释放出的纳米银会和KS发生键合反应生成KS-纳米银复合物,导致纳米银释放出大量的Ag+增加了细胞膜的通透性,从而使得进入细菌内的Ag~+和KS比单独作用时进入胞内的抗菌剂增多,产生更强的抗菌性能,从而表现出协同抗菌效应。本研究基于新型纳米银复合材料与抗生素的联合抗菌性能实验探究了纳米银复合材料与特定抗生素联合用药的最佳组合和相关机制,为今后开发新型抗菌材料提供了新思路并为相关联合用药提供参考。     

Synthesis and antimicrobial activity of esters of 3-ethoxy-4-hydroxybenzaldehyde oxime

A series of oxime esters was synthesized by esterification of 3-ethoxy-4-benzaldehyde oxime with acid chlorides in the presence of triethylamine. They were evaluated for their in vitro antifungal activity against three pathogenic fungi, namely Rhizoctonia bataticola, Fusarium udum, and Alternaria porii, and their antibacterial activity against the three bacterial strains Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumoneae. The compounds bearing medium-length alkyl chains showed greater activity than those with long alkyl chains. An electron-donating group in para position of the aryl ring confers highest activity.     

Synthesis and in vitro biological evaluation of 3, 4-annelated coumarin-pyrimido-pyrimidine systems

3,4-Annelated coumarin-pyrimido-pyrimidine systems (7-imino-8-mercapto-7H-5-oxa-7a,9,12-triaza-benzo[a]anthracen-6-one 2 and 8-hydroxy-7-imino-10-methyl-7H-5-oxa-7a,9,12-triaza-benzo[a]anthracen-6-one 3) were synthesized by the action of 4-amino-2-mercapto-pyrimidine, 2-amino-4-hydroxy-6-methylpyrimidine, respectively, on 4-chlorocoumarin-3-carbonitrile 1. The synthesized compounds were examined for in vitro antibacterial activity against 4 bacterial cultures: Staphylococcus aureus, Streptococcus pneumonia, Escherichia coli, and Aeromonas Salmonicida. The antibacterial activity of synthesized compounds 2 and 3 was compared to antibacterial activity of novobiocin as a standard drug.     

Antimicrobial activities of marine sponges from the Mediterranean Sea

Extracts from marine sponges collected along the French coast of Mediterranean in 1983 have been tested for antibacterial and antifungal activities. The distribution of antimicrobial activities against five classes of microorganisms (5 Gram-positive and 5 Gram-negative terrestrial bacteria, 10 marine bacteria, 15 human pathogenic fungi and 5 phytopathogenic fungi) among 28 species of marine demosponges is reported. Antibacterial activity against Gram-positive bacteria (77%) is much more frequent than against Gram-negative bacteria (53%). Activity against marine bacteria and yeast is less frequent (32%). Those sponges which contain a large quantily of symbiotic bacteria display a weak antifungal activity.     

Antimicrobial activity of Red Sea corals

Scleractinian corals and alcyonacean soft corals are the two most dominant groups of benthic marine organisms inhabiting the coral reefs of the Gulf of Eilat, northern Red Sea. Antimicrobial assays performed with extracts of six dominant Red Sea stony corals and six dominant soft corals against marine bacteria isolated from the seawater surrounding the corals revealed considerable variability in antimicrobial activity. The results demonstrated that, while the majority (83%) of Red Sea alcyonacean soft corals exhibited appreciable antimicrobial activity against marine bacteria isolated from the seawater surrounding the corals, the stony corals had little or no antimicrobial activity. From the active soft coral species examined, Xenia macrospiculata exhibited the highest and most potent antimicrobial activity. Bioassay-directed fractionation indicated that the antimicrobial activity was due to the presence of a range of compounds of different polarities. One of these antibiotic compounds was isolated and identified as desoxyhavannahine, with a minimum inhibitory concentration (MIC) of 48 μg ml⁻¹ against a marine bacterium. The results of the current study suggest that soft and hard corals have developed different means to combat potential microbial infections. Alcyonacean soft corals use chemical defense through the production of antibiotic compounds to combat microbial attack, whereas stony corals seem to rely on other means.     

Bactericidal properties of natural and synthetic sea water as influenced by addition of low amounts of organic matter

The influence of added organic nutrients on the bactericidal capacity of natural and synthetic sea water was investigated, with special reference to levels below the normal concentration of orgnnic matter in natural sea water. Resting cells of Escherichia coli, Staphylococcus aureus and Serratia marinorubra were used as test organisms. The inactivation of test bacteria in synthetic and filter-sterilized natural sea water could be increased by addition of ZoBell's 2216E-broth (containing peptone and yeast extract), peptone or glucose. This effect of the nutrients depended on chemical nature and concentration as well as on specific properties of the test strains. In regard to peptone, less than 10^-4 mg/ml must be added to induce bacterial kill exceeding that observed with unsupplemented controls. With raw sea water, no fundamentally different results were obtained; however, the influence of marine bacteria was clearly discernible. All findings support a previously offered hypothesis of nutrient-dependent antibacterial activity of sea water which is discussed in detail.     

Synthesis,structural characterization and antimicrobial activity of nickel(II) tetrathiocyanato dicuperate(I) complexes with some acyl hydrazones

Heterobimetallic complexes of the type Ni[Cu(SCN)₂]₂ · L (where L = acetophenone benzoylhydrazone, acetophenone isonicotinoyl hydrazone, acetophenone salicyloylhydrazone (ash), acetophenone anthraniloylhydrazone, p-hydroxy acetophenone benzoylhydrazone, p-hydroxy acetophenone isonicotinoyl hydrazone p-hydroxy acetophenone salicyloylhydrazone (phash), p-hydroxy acetophenone anthraniloyl hydrazone), were synthesized and characterized. The complexes are polymeric, insoluble in common organic solvents and are nonelectrolytes. Magnetic moments and electronic spectral studies suggest a spin-free octahedral geometry for the complexes. IR spectra show the bidentate nature of all the ligands bonding through >C=O and >C=N–groups. The SCN group acts as a bridge between two metal centers. X-ray powder diffraction parameters for Ni[Cu(SCN)₂]₂ · ash and Ni[Cu(SCN)₂]₂ · phash correspond to orthorhombic and tetrahedral crystal lattices, respectively, for these complexes. The complexes show a significant antifungal activity against Rizoctonia, Stemphylium and Aspergillus sp. and antibacterial activity against Clostridium and Pseudomonas sp. The metal complexes are more active than the ligands.     

Synthesis and antibacterial activity of a Fe3O4–AgCl nanocomposite against Escherichia coli

In this investigation, Fe₃O₄ magnetic nanoparticles (MNPs) were prepared by the alkalinization of an aqueous medium containing ferrous sulfate and ferric chloride. In the next step, a Fe₃O₄–AgCl magnetic nanocomposite was fabricated by the drop-by-drop addition of silver nitrate solution into a NaCl solution containing Fe₃O₄ MNPs. All prepared nanoparticles were characterized by transition electron microscopy (TEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDS). Both particle types varied in size from 2.5 to 20?nm, with an average size of 7.5?nm for Fe₃O₄ MNPs and 12.5?nm for Fe₃O₄–AgCl nanocomposites. The antibacterial effect of the Fe₃O₄ MNPs and fabricated Fe₃O₄–AgCl nanocomposites against Escherichia coli (ATCC 35218) were investigated by conventional serial agar dilution method using the Müller–Hinton Agar medium. The minimum inhibitory concentration was 4?mg?mL^?1 for Fe₃O₄ MNPs and 2?mg?mL^?1 for the Fe₃O₄–AgCl magnetic nanocomposites. Time-kill course assays showed that the Fe₃O₄–AgCl magnetic nanocomposites successfully killed all inoculated bacterial cells during an exposure time of 60?min. The antibacterial activity of recycled Fe₃O₄–AgCl magnetic nanocomposites over four 60?min cycles of antibacterial treatment was further tested against E. coli by the colony-forming unit (CFU) method. The antibacterial efficiency of the nanocomposites was constant over two cycles of antibacterial testing.     

Unraveling the antibacterial mode of action of a clay from the Colombian Amazon

Natural antibacterial clays can inhibit growth of human pathogens; therefore, understanding the antibacterial mode of action may lead to new applications for health. The antibacterial modes of action have shown differences based on mineralogical constraints. Here we investigate a natural clay from the Colombian Amazon (AMZ) known to the Uitoto natives as a healing clay. The physical and chemical properties of the AMZ clay were compared to standard reference materials: smectite (SWy-1) and kaolinite (API #5) that represent the major minerals in AMZ. We tested model Gram-negative (Escherichia coli ATCC #25922) and Gram-positive (Bacillus subtilis ATCC #6633) bacteria to assess the clay’s antibacterial effectiveness against different bacterial types. The chemical and physical changes in the microbes were examined using bioimaging and mass spectrometry of clay digests and aqueous leachates. Results indicate that a single dose of AMZ clay (250 mg/mL) induced a 4–6 order of magnitude reduction in cell viability, unlike the reference clays that did not impact bacterial survival. AMZ clay possesses a relatively high specific surface area (51.23 m²/g) and much higher total surface area (278.82 m²/g) than the reference clays. In aqueous suspensions (50 mg clay/mL water), soluble metals are released and the minerals buffer fluid pH between 4.1 and 4.5. We propose that the clay facilitates chemical interactions detrimental to bacteria by absorbing nutrients (e.g., Mg, P) and potentially supplying metals (e.g., Al) toxic to bacteria. This study demonstrates that native traditional knowledge can direct scientific studies.     

Impact of conditions of cultivation and adsorption on antimicrobial activity of marine bacteria

The potency of free-living and animal-associated marine bacteria to produce antimicrobial substances has been studied in 491 strains isolated from northern and southern parts of the Pacific Ocean. A total of 26% (126 out of 491) of the strains examined produced antimicrobial compounds against 11 test bacterial strains (TBS) including the fish pathogens Aeromonas hydrophila and Vibrio anquillarum. Antimicrobial substances (AS) produced by marine bacteria were especially active against Staphylococcus epidermidis, Proteus vulgaris, Enterococcus faecalis, and Candida albicans. Twelve strains, isolated from different sources, were chosen as promising candidates, producing a number of AS. Production of AS varied within 24 to 72 h, increasing in a culture medium based on natural sea water with Br-ions, and after attachment to polymeric surfaces. In order to study the influence of adsorption, selected strains with a high potential for antimicrobial production were cultivated on polymeric surfaces with different hydrophobicities and chemical functionalities. These parameters of the surface hydrophobicity (measured by means of water contact angles) and chemical functionality of the surfaces were manipulated using the photo- and thermochemistry of a polymeric system (diazo-naphto-quinone/novolak) commonly used as a photoresistant material in semiconducto-manufacturing. The highest antimicrobial activities occurred on hydrophilic surfaces (standard exposed photoresistant films), whereas the number of attached cells was greater on hydrophobic surfaces, characterized as unexposed resistant films. These results suggest that the chemical nature of induced hydrophilicity may also be a major factor in controlling antimicrobial activity of adsorbed bacteria. Received: 5 March 1997 / Accepted: 24 August 1997     

Inception of formation and early morphogenesis of the bacterial light organ of the sea urchin cardinalfish, <Emphasis Type="Italic">Siphamia versicolor</Emphasis>

The cardinalfish Siphamia versicolor (Perciformes: Apogonidae) forms a bioluminescent symbiosis with the marine luminous bacterium Photobacterium mandapamensis, harboring the bacteria in a ventral, disc-shaped light organ and using the bacterial light apparently for counterillumination and attracting prey. Little definitive information has been available on the developmental and microbiological events surrounding the initiation of symbiosis, a critical stage in the life history of the fish, in S. versicolor or any of the many other species of bacterially luminous fish. To identify the stage at which light organ formation begins, to determine the origin of cells forming the light organ, and to characterize its bacterial colonization status during development, early developmental stages of S. versicolor obtained and reared from wild-caught mouth-brooding males were examined with histological and microbiological methods. A light organ primordium was not evident in embryos, post-embryos, or pre-release larvae, whereas the light organ began to form within 1 day of release of full-term pre-flexion larvae from the mouths of male fish. Analysis of post-release larvae revealed that the light organ arises from a proliferation and differentiation of intestinal epithelial cells, and that it quickly develops structural complexity, including the formation of chambers and gaps contiguous with the intestinal epithelium. However, the nascent light organ remained uncolonized by the symbiotic bacteria through several days of post-release development, even in the presence of high numbers of the symbiotic bacteria. These results demonstrate that the inception of light organ formation in S. versicolor occurs independently of its symbiotic bacteria and that receptivity to bacterial colonization apparently requires substantial post-release development of the light organ. Larvae therefore most likely acquire their symbiotic bacteria from seawater, during or shortly after the transition from the pre-flexion to the flexion developmental stage.