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.

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:

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.

It was at the spur of the moment that we ventured to write something about pheromone attraction. Such is the amount of matter that is available on pheromone attraction.

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.

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.

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

Table 1. List of Seaweeds species collected for the present study SPECIES NAME FAMILY Halimeda gracilis Chlorophyceae Ulva lactuca Chlorophyceae

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

Understanding the various varieties of perfume can help to cut through the difficulty in selecting a fragrance. With so many variances within each category, choosing exactly the right aroma for your body or anyone else's for that matter, is no easy task.

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

Caulerpa racemosa Chlorophyceae Dictyota dichotoma Phaeophyceae Turbinaria ornata Phaeophyceae Padina gymnospora Phaeophyceae Sargassum cinearifolium Phaeophyceae

This champion, this Hero, this version of you, must slay many dragons in your quest. You must confront formidable enemies. Climb perilous peaks. Cross frothing oceans. Must drive your sword deep into the heart of ferocious, incredible beasts. And brave the dangers all alone. You must do these because there is no other way. It is your quest, not another's. And if you are not prepared to do these things, do them mightily, and succeed, then you cannot have this Lover. She will slip onto the lips of another. You will be led to exclaim, "I did not love her well. I did not love her enough." And you will cry until your soul is empty.

Colony morphology, Gram staining, Motility test, Oxidase test, Catalase test, Indole Production, Methyl red test, Voges Proskauer test, Citrate Utilization test, Triple sugar Iron test, Nitrate reduction test, Lactose fermentation, Urease test

No, it is only that her Hero sees in her beautiful sleep what no others can see. It is your ability to follow the lead of her loving fragrance, and count all the challenges to kiss her as mere distractions and delays until you can lay your lips upon hers and pull her to full awareness. It is this enduring quality that she seeks, because she intends to love you long, and love you well, and nothing short of this will enable you to stay the course and give her all she desires. For want of this, she chooses to sleep, resting her energies for the passionate embrace she intends to fill her lifetime. Is there any other reason to stay awake?

In this experiment among 17 Producer and Inducer strains only 3 of them have responded to the quorum sensing principle. (BrB+/BrB-), (GcC+/GcC) and (SA+/SA-)

Mckenney, D., K. E. Brown, and D. G. Allison. 1995. Influence of Pseudomonas aeruginosa exoproducts on virulence factor production in Burkholderia cepacia: evidence of interspecies communication. J. Bacterial 177: 6989 - 6992.

ABSTRACT Bacterial biofilms are sessile communities with high cell density that are ubiquitous in natural, medical, and engineering environments; they are fascinating since they are primitive tissues with an advanced chemical communications network. Currently, there is an explosive amount of biofilm research, most of it with the ultimate aims of biofilm prevention, control, or eradication.The present investigation was aimed to study the cell signaling principle among seaweed epibiotic bacterial organisms , totally 54 bacterial isolates were made out of 20 seaweed species. Among 54 bacterial isolates 17 of them producer, another 17 of them inducer strains rest of 20 isolates were normal that have not showing any signs of activity. The 17 producers and 17 inducer strains were subjected to the cross species induction by quorum sensing principle and found that only 3 of them have responded to the cross species induction in production of antibacterial compound against the respective inducer strains. All the 3 producers and inducer were identified by biochemical analysis and surprisingly that all the 3 producer strain belongs to Genus Pseudomonas and inducer strains were belongs to Genus Vibrio. The supernatant obtain from mixed culture of Pseudomonas & Vibrio shows the antibiotic activity against the various Vibrio species which were isolated from other sources. Such as Vibrio from primary film, Vibrio from sediment Vibrio from seaweeds epibiotics. The obtain results clearly shows the particular Pseudomonas from seaweed epibionts has the capability of producing potential antibiotic compound against wide range of Vibrio species through quorum sensing.

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

Burgess, J. G., H. Miyashita, H. Sudo, and T. Matsunaga.1991. Antibiotic production by marine photosynthetic bacterium Chromatium purpuratum NKPB 031704: Localization of activity to the chromatophores. FEMS Microbiol. Lett. 84: 301 - 306.

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.

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.

A sleeping Lover lays at the core of many timeless legends. A kiss, or magic potion is needed to awaken her. To grasp the eternal truths displayed so beautifully in these old texts, you need only to see it is not a story. It is instruction to you. It is your guide, your encouragement, your path to be a Hero.

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

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

*Corresponding author e-mail: vijay10bas@yahoo.co.in INTRODUCTION We are living in a microbial planet. About 71 % of the surface of this planet is covered by sea water. A typical milliliter of seawater contains 103 fungal cells, 106 bacteria, and 107 viruses, including pathogens that cause widespread mortalities and microbes that initiate fouling of host surfaces (Rheinheimer, 1992). Thus, marine plants and animals are continually exposed to high concentrations of potentially harmful microbes. These microorganisms in nature exists as free living planktonic mode of life in sea water or it may exist as epibiotic organisms in various living and nonliving surfaces. Among living organisms, seaweeds and invertebrates act as suitable substrate for the establishment of epibiotic organisms Seaweeds are known to release a large amount of organic carbon into the surrounding environment providing a nutrient rich habitat for microorganisms like bacteria. Bacteria are generally considered to be independent unicellular organisms. One cell accomplishes all of the tasks of feeding, locomotion, 'reproduction, respiration and all other processes necessary to keep an organism alive. There are several classes of bacteria such as primary film forming bacteria, sediment bacteria, symbiotic bacteria, and epibiotic bacteria in various aquatic organisms. The marine surface environment is a site of intense composition for living space by a wide variety of organisms. Bacteria are generally recognized as primary colonizers of this habitat and are able to form biofilm on marine surface such as invertebrates and algae (Bryers, et al., 1982). Bacteria may also be abundant on the surfaces of some algae as an important epibiotic organism. In many cases, the bacterial population found to be specific, with changes occurring throughout the year or life span of the algal surface. This algal-bacterial relationship is symbiotic in most cases; the epibiotic bacteria in seaweed play a protective role by releasing secondary metabolites into the surrounding seawater that help preventing extensive fouling of the surface. Epibiotic bacteria are therefore attracting attention as a source of new natural products. Bacteria from the larvae of some crustaceans protect them from fungal infection by the production of simple antimicrobial compounds. Bacteria isolated from the surface of a tunicate prevented the settlement of barnacle and tunicate larvae exposed to the bacteria as biofilm in petridishes (Evelyn et al., 2001).

It was at the spur of the moment that we ventured to write something about pheromone attraction. Such is the amount of matter that is available on pheromone attraction.

Icebreaker Pheromone Cologne

b) Among these 17-inducer strain 6 strains were isolated from Hypnea musiformis, 6 from Gracillaria edulis, 4 from Ulva lactuca & 1 from sediment.

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.

Zhang, L., P. J. Murphy, and A. Kerr. 1993. Agrobacterium conjugation and gene regulation by /V-acyl-L-homoserine lactones. Nature 362: 446 - 448.

GcA-, GcB-, GcC-, GcD- GcE-, GcF- Gracillaria edulis U1-, U2-, U3-, U4- Ulva lactuca SA- Sediment

Armstrong, E., J. D. Mckenzie, and G.T. G. Worthy. 1999. Aquaculture of sponges on scallops for natural products research and antifouling. J. Biotechnol. 70: 163-174. 70. 21-

illaud, A. C, and B. Austin. 1990. Inhibition of the fish pathogen, Serratia liquefaciens by an antibiotic producing isolates of planococcus recovered from sea water. Journal of

a) Among these 17 producers strain 6 strains were isolated from Hypnea musiformis. 6 from Gracillaria edulis, 4 from Ulva lactuca & 1 from Sediment.

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."

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

Pursuing the Scent of the Sleeping Lover So, the Hero who seeks to awaken the soul of the Lover must be well prepared. You must prepare your body for the rigors of prolonged loving. You must prepare your soul to reveal all, to give all, to allow it to be encircled. To be consumed again and again. This is the champion's quest, to prepare for a life of giving. To clear your mind of every obstacle, to rid your life of every hindrance, to tirelessly, single-mindedly pursue the scent of the sleeping Lover until you are lip-to-lip.

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.

Liquid Trust Liquid Trust contains oxytocin - hormone that controls the level of trust in people. Oxytocin makes people more willing to bond with others. Liquid Trust Body Spray is created to increase trust in the wearer. This product is applied every morning after showering along with a favorite cologne or perfume.

Dekievit, T. R., and B. H. Iglewski. 2000. Bacterial quorum sensing in pathogenic relationships. Infect. Immun. 66: 4839-4849. Dimango, E., H. J. Zar, R. Bryan, and A. Prince. 1995. Diverse Pseudomonas aerugniosa gene products stimulate respiratory epithelial cells to produce interleukin-8. J. Clin-lnvestig. 96: 2204-2210.

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.).

Whiteley, M., K.M.Lee, and E.P.Greenberg.1999. Identification of genes controlled by quorum sensing in Pseudomonas aeruginosa. Proc.Natl.Acad.Sci.USA.96:13904-13909.

Awaken The Lover Who awakens the soul of the Lover? Who has the magic potion to open her eyes wide and seek her consuming embrace? What first put the Lover to sleep? Where was the magic potion hidden? Who chose this Hero to apply it?

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

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.

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

U1+, U2+, U3+, U4+ Ulva lactuca SA+ Sediment INDUCER STRAIN BrA-, BrB-, BrC-, BrD-, BrE-, BrF- Hypnea musiformis

Chypre is another popular blend, typically a soft aroma of outdoor woody-leather scents mixed with citrus aromas. These blends are typically easy to spot when shopping as they are almost always encased in leather as a protective covering. The mixing of fragrances has shifted toward the fruity or "green" scents and leaves behind a gentle musk.

Holmstrom, C, D. Rittschof, and S. Kjelleberg. 1992. Inhibition of settlement by larvae of Balanus amphitrite by a surface - colonizing marine bacterium. Appl. Env. Microbiol. 58: 2111 -2115.

The most popular based on sales is the floral category. Floral scents are typically a varying concoction of flower aromas blended together for a unique scent. Jasmine, gardenia, and rose bases are the most popular of the popular, often for their unique blending style and their ability to match most body chemistries. Floral scents can vary from very strong to very mild.

Trischman, J.A., D.M.Tapiolas, P.R.Jensen, R.Dwight, W.Fenical, T.C.Mckee, C.M.Ireland, T.J.Stout, J.CIarely. 1994. Salinamide - A and salinamide - B - anti -inflammatory depsipeptides from a marine streptomycete. J.Am.Chem.Soc. 116: 757 -758.

Hypnea musiformis 1. BrA+ 2. BrB+ 3. BrC+ 4. BrD+ 5. BrE+ 6. BrF+ BrA- BrB- BrC- BrD- BrE- BrF- Br A+ /Br A-

The Quest for Fulfillment The quest of each individual, in this modern age, is arise out of the ashes of ancient myth, to slough off today's culture of "trying"; to turn sharply away from the lies of an under-achieving culture; to find the rough, perilous way to your soul. For it is the satisfaction of your soul that you are missing. No amount of "trying to make it happen" is going to get you there. Only finding the heroic core buried in your soul, your reason for existence, your drive to be a true Hero, can set you on the path to fulfillment... your soul desperately seeks you to touch this sleeping Lover.

In the present investigation, it was aimed to produce the antibiotics from the seaweed epibionts through quorum sensing principle. The bacterial isolates of seaweed epibionts were identified as species of Pseudomonas and Vibrio from seaweeds Hypnea musiformis and Gracillaria edulis. In this study the Pseudomonas acts as a producer strain and Vibrio as inducer strain. The recent finding says that the seaweed epibionts having potential to control the metabolic activity of competitor organisms. Allison et al., 1998 reported that many bacterial strains up on attaching to a surface produce exopolysaccharides or exopolypeptides. In addition, it has been postulated that exopolysaccharides could mediate the attachment of the bacteria to the surface and induce metabolic changes.

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.

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

All the 17 strains were named as PRODUCERS STAINS BrA+, BrB+, BrC+, BrD+, BrE+, BrF+ Hypnea musiformis GcA+, GcB+, GcC+, GcD+, GcE+, GcF+ Gracillaria edulis

The earlier genetic analysis in Pseudomonas reveals the Pseudomonas consist of two quorum sensing systems as Las R1-I and Rh1R-l and have linked with R and I genes, in addition recently a third Lux R homolog that is advanced to a cluster of quorum sensing - controlled (qsc) genes were detected. Las R is a transcriptional regulator that responses primarily to the Las I - generated signal and Rh1R is a transcriptional regularly that responses best to the Rhl -generated signal. In Pseudomonas auriginosa, at low population densities Las I produce a basel level of 3-O-C12-HSL. As density increases, 3-0-C12-HSL builds to a critical concentration, at which point interacts with LasR. This Las R -3-0-012-HSL complex that activates transcription of a number of genes [Whileley, et al., 1999].

?? 2007 Boyd L. Jentzsch

We hope that through reading of this article on pheromone attraction, interest in pheromone attraction is once again activated.
Author Dave Thomson

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