Interference With Cell-to-cell Signaling: a Potential Therapeutic Approach Against Vibrio Species 

Vijaya baskar .P 1* AND Veera ravi.A 2 1. Department of biotechnology Dr. G.R.D.C.S, Coimbatore. 2. Department of biotechnology, Alagappa University, karaikudi.

Evelyn, A., Y. Liming, K.G. Boyd, P. C. Wright, and J. G. Burgess. 2001. The symbiotic role of marine microbes on living surfaces. Hydrobiologia. 461: 37 - 40

Dictyota batryensis Phaeophyceae Sargassum sps Phaeophyceae Hypnea musciformis Rhodophyceae Acanthophora dendroides Rhodophyceae Jania rubens Rhodophyceae

It was with keen interest that we got about to writing on attraction pheromone. Hope you read and appreciate it with equal interest.

Mearns - Spragg, A., K. G. Boyd, and M. O. Hubble. 1997. Antibiotics from surface associated marine bacteria. Proceedings of the fourth underwater science symposium. The society for underwater Technology, London: 147 - 157.

Burgess, J. G., M. Elizabeth, M. Bregu, A. Meams-Sprugg, and K. G. Boyd. 1999 Microbial antagonism: a neglected avenue of natural products research. J. Biotechnology. 70. 27-32.

Lemos, M. L, A. E. Toranzod, and L. J. Barja. 1986. Antibiotic activity of epiphytic bacteria isolated from intertidal seaweeds. Microbial Ecology. 11: 149-163.

This is the secret world of the undisclosed Lover. This is the place the Lover has chosen to launch her crusade to catch eternal love from the clutches of the ordinary world. She has chosen to sleep in this world for there is none within it to entice her to stay. But with her sleep she knows her sleeping pheromones will arouse only those most worthy of its essence, most able to arouse her and consummate the reason for her long sleep.

All the above mentioned biochemical tests were performed by following standard methodology given in the Microbiological Laboratory Manual by James 3.Cappuccino (1999).

Table 2: The results of Quorum Sensing analysis of epibiotic bacterial isolates from seaweeds. Seaweed sample Producer organism Inducer organism Cross-species producer with inducer Cross-species supernatant test with inducer Zone of clearance (mm)

Allison, D. G., B. Ruiz, C. Sanjose, A. Jaspe, and P.Gilbert. 1998. Extracellular products as mediators of the formation and detachment of Pseudomonas fluorescens biofilms. FEMS Microbial Lett. 167: 179-184.

In this present work the totally 54 isolates were screened from 20 different seaweed species out of which 34 species were showed the signs of quorum sensing i.e. 17 producer and 17 inducer strains, but when these organisms where subjected to quorum sensing principle in mixed culture only 3 them have responded. So the present study reveals around 17% of bacterial species isolated from seaweeds and sediment were responded to quorum sensing. According to the results of bacteria isolated from marine algae surfaces indicated that the incidence of antibiotic producing strains from this habitat was 20% where as that from sea water was only a few percent. In the present study also reveals more or less the same ratio in Pseudomonas spp. was observed. Our results also reveals a results of Kell et al., 1995; Stead et al., 1996; they have said the culture supernatant of Pseudomonas sps known to contain AHLS which induces the production of phenazine antibiotics. In this investigation due to time constraint, It was not attempted to identify the active compound secreted by Pseudomonas through quorum sensing, which may leave the space for the further intensive research in future.

Vandevivere, P., and D. L. Kirchman.1993. Attachment stimulates exopolysaccharide synthesis by a bacterium. Appl. Environ. Microbiol. 59: 3280-3286.

Then the quest changes, for the energy expended thereafter is to keep lips locked together, never allowing a sliver of light to come between them. This is the new, and greater quest. For, winning the heart of a sleeping Lover is one thing. Keeping the heart of an awakened Lover quite another. And if you are not prepared for this new consuming task before you begin, you will have scant time for reflection to do it once you have slipped endlessly into her soul. The Hero must prepare diligently. Or extinguish all hope of merging with the awakened, fully aware Lover.

Liming, Y., G. K. Boyd, and J. G. Burgess. 2002. Surface induced attachment induced production of Antimicrobial compounds by marine epibiotic Bacteria using modified Roller Bottle. Mar. Biotechnology. 4: 356 - 366.

One of the more musky yet strong aromas falls in the leather category. Leather fragrances have almost an animal like smokiness to them that is suitable for men and some women, although it is not recommended to purchase leather fragrances as a gift for women since it matches very few female pheromones. The leather scents are reminiscent of the aroma that tanning animal hides gives off. While it is not a fragrance for everyone, those for whom it works for creates a very original scent that can be very attractive.

This experiment does not showed any inhibition zones Bacterial identification The 3 producer and 3 inducer strains which were responded the quorum sensing principles alone were subjected to biochemical analysis for identification. The obtained results revealed that all the producer strains showed sings of Pseudomonas sps and the inducer strains showed signs of Vibrio sps. So, based on the obtained result all the producer strains seems to be a Pseudomonas sps where as all the Inducer strain belongs to the genus vibrio.

Kell, D. B., A. S. Kaprelyants, and A. Grafen. 1995. Pheromones, Social behaviour and the functions of secondary metabolism in bacteria. Trends in Ecol. and Evol. 10: 126-129.

EXPERIMENT NUMBER 2 In this experiment, culture supernatant was obtained as per the procedure given in the experiment 1. 50ml of supernatant was mixed with equal volume of 80% methanol and 1% acetic acid mixture and it was shaked thoroughly in a separating funnel. Finally the methanol and acetic acid fractions were collected and concentrated by evaporation using water bath at 55??C. The viscous colloidal residues were resuspended in 600 microlitre of 50% methanol and it was used for antibacterial assay against different test organism.

All the cultures were incubated at 28??C for 5 days. After the incubation period the cultures were centrifuged at 10,000 rpm for 15mins. The supernatant was collected and subjected to antibacterial assay with respective inducer strain.

Strip away the allusions to sensuality in these old stories. They are the stories of the culmination of the Hero Quest. So, listen carefully and hear your quest, your future revealed herein.

In gram positive bacteria peptide and derivative peptide based signaling molecules seem to be the predominant mode of communication. During high cell density the marine bacteria can produce enzymes, surfactants, toxins, and antibiotics by the chemical signal communication. Marine epibiotic bacteria are also known to produce compounds active against drug resistant hospital pathogen by the cross species induction method. Building on assays described by Austin (Billaud and Austin 1990) a screening procedure has been developed in which marine bacteria are challenged by exposing them to terrestrial bacteria prior to assay of antimicrobial compounds. Hence in this present investigation it is proposed to find out the ability of sea weed epibiotic bacterial organism to produce antibacterial compounds through quorum sensing.

Seaweeds itself secretes secondary metabolites to prevent fouling and grazing. In addition to that epibiotic bacteria on macro algae can also produce antifouling compounds that work in concert with the seaweed derived compounds to protect the seaweed surface. Recent studies have highlighted important roles of epibiotic bacteria colonizing the surface of seaweeds and releasing antifouling compounds. For the past 50 years antibiotics have revolutionized medicine by providing cures for formerly life threatening diseases. However, strains of bacteria have recently emerged that are virtually unresponsive to antibiotics such multidrug resistance, arising mainly through antibiotic misuse, is now recognized as a global health problem. In this situation, it is clear that new classes of antibiotics are urgently needed. Many marine bacteria have been shown to produce secondary metabolites that display antibacterial properties. The first antibiotic from a marine bacterium was identified and characterized in 1966. In addition, bacteria in biofilm on the surface of marine organisms have been documented to contain a higher proportion of antibiotic producing bacteria than some other marine environments (Burgess, et al., 1999). Marine epibiotic bacteria, associated with nutrient-rich algal surfaces have also been shown to produce antibacterial secondary metabolites which inhibit the settlement of potential competitors. Recently a lot new novel antibiotics such as Phenazine, thiomarinol, phenazine-1-carboxylic acid, 1-hydroxyphenazine 2-n-heptylquinol-4-one, 2-n-nonylquinol-4-one pyolipic, loloatins, agrochelin, sesbanimides, pelagiomicins, indomycione and indomycione have been identified from various marine epibiotic bacterial organisms. In particular, some species of the genus Pseudomonas produce both antibiotics and several other bioactive substances. For example, Pseudoalteromonas rubra and Pseudoalteromonas aurantia have been reported to be antibiotic producing bacteria. The phenomenon of higher organisms utilizing their associated microflora for the production of beneficial secondary metabolites is common in the marine environment (Yotsu, et al., 1987). A study of bacteria isolated from marine algae surfaces indicated that the incidence of antibiotic producing strains from this habitat was 20% whereas that from sea water was only a few percent. In addition, some bacteria that previously did not produce any active compounds have been found to be producing such metabolites when they are exposed to other bacterial species or extra cellular chemical from other bacteria. Bacteria may also produce antimicrobial compounds when they sense the presence of competing organisms. However, few attempts have been made to study such chemical communication between different bacterial species or how this might affect. The secretion of antimicrobial compounds (Mearns-Spragg, et al., 1998). Bacterial communication by the chemical signals for specific function is simply known as Quorum sensing. In which a bacterial population receives input from the environment and elicits an appropriate response (Hiroaki and Kristina. 2003). The term "quorum sensing" describes the ability of a microorganism to perceive and response to diffusible signal molecules. Bacterial cells sense their population density through a sophisticated cell to cell communication system and trigger expression of particular genes. Tne first system of density-dependent regulation was studied in detail with the luminescence of Photobacterium fischeri (formerly known as Vibrio fischeri) by Bassler et al., 1997. Eventually, they discovered that 3-oxo-N-(tetrahydro-2-oxo-3-furanyl) hexanamid or N-3-(oxohexanoyl) homoserine lactone (OHHL) was responsible the agent in the broth that induced luminescence. Followed by this many researchers have confirmed that in Gram negative bacteria acyl-homoserine lactone is responsible for the cell to cell communication system.

It was with keen interest that we got about to writing on attraction pheromone. Hope you read and appreciate it with equal interest.

Icebreaker Pheromone Cologne

The Oriental aromas are more balsamic with a hint of oak moss, animal scents, or vanilla which creates a noticeable lingering in the air. Obsession is a very popular brand of an Oriental based perfume. Oriental perfumes are considered quite full if not even "heavy."

Bainton, N. J., P. Stead, and S.R. Chhabra. 1992. N-(3-oxohexanoyl)-L-homoserine Lactone regulated carbapenem antiobiotic production in Erwinia carotovora. Biochem. J. 288: 997-1004.

c) The normal 20 bacterial strains isolated from 20 algal species were crossed with terrestrial bacteria such as E-coli, Staphylococcus aureus separately

NIL NIL NIL NIL Gracillaria edulis 7. GcA+ 8. GcB+ 9. GcC+ 10. GcD+ 11. GcE+ 12. GcF+ GcA- GcB-

Br B+ /Br B- Br C+/Br C- Br D+/Br D- Br E+/Br E- Br F+/Br F- BrA- Br B- BrC- BrD- BrE- BrF- NIL 39

Liming, Y., K. G. Boyd, and D. R. Adams. 2003. Biofilm specific cross species induction of antimicrobial compounds in Bacilli. AppI.Envi.Microbiol, 69:3719-3727.

Primal Instinct This is a very strong androstenone product. There is greater risk of overdose, so it is advised not to apply too much of this product. Very often, even one drop is too much for some men. There are two versions of this oil-based pheromone product - scented and unscented. Users should be aware that this product is not recommended for young men.

Yotsu, T., Y.Meguro, A. Endo, M. Murate, H. Naold, and T. Yasumoto.1987. Production of tetrodotoxin and its derivatives by Pseudomonas sp. Toxicon 25: 225 - 228

GcC- GcD- GcE- GcF- Gc A+/GcA- Gc B+/GcB- Gc C+/GcC- Gc D+/GcD- Gc E+ /GcE- Gc F+/GcF- GcA- GcB- GcC-

Hiroaki, S., and K. M. Smith. 2003. Molecular mechanisms of bacterial quorum sensing as a new drug target. Curr. Opin. Chem. Biol. 7: 586-591. Hoang, T., Y. Ma, R. J. Stern, M. R. Meneil, and H. P. Schwezer. 1999. Construction and use of low-copy number T7 expression vectors for purification of problem proteins: purification of Mycobacterium tuberculosis RmID and Pseudomonas aeruginosa Lasl and Rhll proteins, and functional analysis of purified Rhll. Gene. 237: 361 - 371.

Hypnea valentiae Rhodophyceae Hypnea pannose Rhodophyceae Hypnea esperi Rhodophyceae Acanthophora spicifera Rhodophyceae ISOLATION OF EPIPHYTIC BACTERIA

Patterson, G. L., and C. Ml. Bolis. 1997. Fungal cell - wall poly sacchraides elicit an antifungal secondary metabolite in the Cyanobacterium Scytonema ocellatum. J. Phycology. 33: 54 - 60.

You must be one with your quest, your ardent dream. There can be no separation between it and you; none whatsoever between life and Hero Quest. The perilous journey IS life. It is the arduous journey, the heroic tasks you must conquer to have it. Or you will not have life at all.

Rheinheimer, G.1992.Aquatic microbiology (Wiley, New York), 3rd Ed. Stead, P., B. A. M. Rudd, H. Bradshaw, D. Nobel, and M. J. Dawson. 1996. Induction of phenazine biosynthesis in cultures Pseudomonas aeruginosa by L-N-(3-oxohexanoyl) homoserine lactone. FEMS Microbiology Letters 140: 15-22.

The Hero she seeks must prepare himself for her awakening long before the opportunity to win her wafts into the air. It is not his fortune that she seeks. It is not his beauty - for her Hero, her champion, will always be beautiful to her, no matter his visage. It is not his courage to best all in his quest to find her that underlies her dormant love.

In the gram negative bacteria AHSL is an active principle of quorum sensing. Our producer strain is also been identified as Pseudomonas sps. So in these organisms also active principle must falls under the AHSL. The cell-cell signaling mechanism can either require import of the signal and subsequent interaction with intracellular effectors or a two-component signaling system that transducers the information across the membrane. In V. harveyi genetic analysis of the density sensing apparatus has two independent density-sensing systems, and each is composed of a sensor-auto inducer pair; system one is composed of sensor I and Al -1, and system two is composed of sensor 2 and AI-2. The two densities - sensing system are redundant, because a null mutation in either system alone results in expression of hidden genes (Bassler, et al.,1999.).

MATERIALS AND METHODS: SAMPLE COLLECTION: Seaweed samples were collected from Gulf of Mannar Marine Biosphere Reserve and identified up to species level by using CMFRI bulletin (14) as follows:

Once awake, she will be demanding. She will want all of your body. Nothing less. She will want every part of your soul. She will consume your body and soul, then she will ask for more. More still. More again. And if you, the Hero, the enduring champion cannot quench her hunger, and that thrice daily, she will find another. Or sleep once again.

Bassler, B. L., E. P. Greenberg, and A. M. Sterens. 1997. Cross species induction of luminescence in the quorum - sensing bacterium Vibrio harveyi. J. Bacteriology. 179: 4043-4045.

The citrus family of perfumes is, as expected, a fruity scent derived from natural fruit oils. Pomello and lemon and orange and even grapefruit are among the most popular bases for citrus perfumes. Typically younger individuals prefer the citrus based over most other fragrances.

BACTERIAL IDENTIFICATION The organisms responded to the quorum sensing process alone were identified by the following biochemical analysis.

Wagner, V. E., D. Bushneil, and L. Passador. 2003. Micro array analysis of Pseudomonas aeruginosa quorum sensing regulons. J. Bacteriol, 185: 2080 - 2095.

U2- U3- U4- U1+/U1- U2+/U2- U3+/U3- U4+/U4- U1- U2- U3- U4- NIL NIL NIL NIL Sediment 17. SA+ SA- SA+/SA- SA- 28

Quorum sensing usually focused on the bacteria growing in homogeneous environment. However few studies have attempted to a study this principle in heterogeneous environment also. In this present investigation we have attempted to study both homogeneous as well as heterogeneous environment. In former one we have isolated producer strain in seaweed eipbionts and it shows inhibitory activity against the inducer organism at the same seaweed epibionts. Later producer strains from seaweed epibionts, were treated with various Vibrio organisms from different environment. The obtain result of this study shows that the producer strain are capable of secreting antibiotic compounds not only to their natural competitors in its own habitate but also to the pathogen inhabiting in a distant related environment.

Boyd, K.G., A. Mearns -Soragg, G. Briedley, K. Hatzidimitriou, A. Rennie, M. Bregu, M. 0. Hubble, and J. G. Burgess. 1998. Antifouling potential of epiphytic marine bacteria from the surface of marine algae. Plouzane, France: 128 -136.

TEST ORGANISMS: 1. Epiphytic Vibrio from seaweeds 2. Vibrio from primary film 3. Vibrio from Sediments 4. Pathogenic bacteria such as Escherichia coli, Staphylococcus aureus, Salmonella sp. and Proteus sp

Fougere is a French fragrance that has gained huge popularity in the United States. Literally translated it means "fern," and has an interested fragrance blend of musty moss-like aroma with herbs such as lavender. Men have a particular fondness for this perfume for its subtle blended scent.

The results of Vanderivere and Kirchman 1993 suggest that the addition of increased surface by adding sand will induce the exopolymer synthesis through the high cell density dependent system. In the same way the bacterial organisms attached to the surface of seaweed shows alteration of genes expression may be due to the response to the high competitive environment. When cell density increases the competition for space and nutrients are also increased. So the existing bacteria were forced to protect themselves in this competitive environment. Normally in this condition the bacteria will be activated to induce the expression of certain hidden genes in genetic material through quorum sensing. The quorum sensing is principles were active compounds (autoinducer) from bacterial cell will promote the expression of a particular hidden gene of other bacterial organism in a stressed condition.

The collected seaweed samples were thoroughly washed with sterile seawater to removes the loosely attached bacteria/particles. Seaweed fronds were scrubbed with sterile cotton swabs to obtain epiphytic bacteria. Epiphytic bacterial organism in the swab were inoculated in sterile peptone broth (50% sea water) and incubated at 28??C in an incubated shaker (220 rpm / min) for overnight. After the incubation period the enriched cultures were serially diluted up to 10-8 concentration and 200 microlitre of each diluted samples were transferred into the nutrient agar plate (50% sea water). The plates were incubated at 28??C for 5 days and the plates with crowded colonies were selected. In the crowded plates those colonies, which showed the sign of inhibition zone around its margin to the neighboring colony, were selected and considered as producer strain. The neighboring sensitive colonies were treated as inducer strain. Both producer and inducer strains were streaked repeatedly until to get pure culture. The pure culture were properly labeled and subjected to the quorum sensing analysis.

?? 2007 Boyd L. Jentzsch

It is with a heavy heart that we have come to the end of this beautiful composition on attraction pheromone. Please do disburse its beauty to others.
Author Dave Thomson

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