10/23/2018

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.


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

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.
 




When a child shows a flicker of understanding when talking about homemade pheromones, we feel that the objective of the meaning of homemade pheromones being spread, being achieved.

Scientists believe that animals (usually the females) such as insects and mammals send out these chemical signals to tell the male of their species that their genes are different from theirs. This gene diversity is important in producing offspring with better chances of survival. The perfume industry has capitalized on pheromones as a means to increase one's sexual attractiveness to the opposite sex. Animals such as the whale and the musk deer were hunted down for these chemicals.

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.

Meains - Spragg, A., M. Bregu, K. G. Boyd, and J. G. Burgess. 1998. Cross species induction and enhancement of antimicrobial activity produced by epibiotic bacteria from marine algae and invertebrates, after exposure to terrestrial bacteria. Letters in Applied Microbiology. 27: 142-146.

Today's fervor for environmental preservation has also hit the medical industry as researchers, scientists and doctors are discovering the importance of the newest emerging field called environmental endocrinology. Doctors are now learning how environmental endocrinology, or the effect of daily stressors like light, food and crowding on multiple endocrine systems, controls the rate of aging and the quality of life. This also covers reproductive endocrinology, converging to become what we call menopause medicine.

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

Calendar consciousness first developed in women because their natural body rhythms corresponded to observations of the moon, and as it turns out, 28 days is not the true average of the natural female cycle. In ancient mythologies it is clearly related to the full moon, but in the modern world the female cycle is disturbed by what some researchers believe to be the existence of artificial lights and the use of artificial hormones. Many light sources including TV and computer screens, have probably perturbed the female cycle, and shortened it.

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.

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

GcD- GcE- GcF- NIL NIL 26 NIL NIL NIL Ulva lactuca 13. U1+ 14: U2+ 15. U3+ 16. U4+ U1-

Does personality figure in sexual attraction? Yes, but so does your perception of a potential mate's personality. According to a research conducted by Klohnen, E.C., & S. Luo in 2003 on interpersonal attraction and personality, a person's sense of self-security and at least the person's perception of his/her partner were found to be strong determinants of attraction in hypothetical situations. What does this tell us? We prefer a certain personality type, which attracts you to a person. But aside from the actual personality of the person, which can only be verified through close interaction through time, it is your perception of your potential partner that attracts you to him/her, whether the person of your affection truly has that kind of personality or not. This could probably account for a statement commonly heard from men and women on their failed relationships: "I thought he/she was this kind of person."

Doctors who are studying environmental endocrinology are in the vanguard of an elite group of forward-thinking physicians and researchers trying to put the scientific method back into medicine, spearheaded by a researcher named T.S. Wiley.

In concluding this discussion, the quorum sensing is wider spread among bacterial population then was previously thought, (In Gram positive, Gram negative bacterial communication). Current assays for antimicrobial activities are inadequate because some antibiotic producing bacteria may require the presence of another bacterial species. These findings have important implication for the discovery of novel antimicrobial compounds from marine bacteria and may allow the development of new methods for screening novel compounds active against multidrug resistant bacteria.

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.

RESULTS AND DISCUSSION: QUORUM SENSING/CROSS SPECIES INDUCTION ANALYSIS In the present investigation totally 54 isolates were collected out of seaweed species. Among 54 isolates, 17 of them are producer strain, another 17 are the inducer strain rest of 20 isolates is normal and not showing any signs of activity (Table.2).

"Opposites attract" is a law of attraction, at least where electromagnetism is concerned. But are there laws about attraction between two people? "In a world that is full of strangers" as a line in a famous song of the 1980's goes, is there a clear set of rules that allows two people to fall for each other?

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.

In culture system B 200ul of 16 hours old producer strain alone was inoculated. In culture system C 200ul of 16 hours old inducer strain alone was inoculated.

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

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

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.

So should I stop trying to become attractive? More than trying to become physically attractive, work on all aspects of your health: physical, emotional, mental, and spiritual. Physical attraction is still a precursor. Remember, biology predisposes us to choose the partner with the healthiest genes. Where your emotions are concerned, just ask this to yourself: would you want to spend time with a person who feels insecure about him/herself?

When a child shows a flicker of understanding when talking about homemade pheromones, we feel that the objective of the meaning of homemade pheromones being spread, being achieved.



Icebreaker Pheromone Cologne



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-

ANTIBIOTIC ASSAYS Antibiotic activity was performed in duplicate using a standard paper disc diffusion method as well as well assay. In well assay 10mm in diameter wells were made in marine agar plates and the plates were swabbed with 16 hours old inducer strain. To these wells 200ul of cell free supernatant were added to each well. In paper disc assay the Watmann no.1 filter paper discs (6mm in diameter) were saturated with 200ul of cell free supernatant. The impregnant discs were Dlaced in the centre of the plates swabbed with test organisms. The plates were Incubated at 37??C overnight and observed for inhibition zone. The zone of inhibition was measured as the distance from the border of paper disc to the edge of the clear zone and expressed in mm.

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

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.

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.

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

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

Does personality figure in sexual attraction? Yes, but so does your perception of a potential mate's personality. According to a research conducted by Klohnen, E.C., & S. Luo in 2003 on interpersonal attraction and personality, a person's sense of self-security and at least the person's perception of his/her partner were found to be strong determinants of attraction in hypothetical situations. What does this tell us? We prefer a certain personality type, which attracts you to a person. But aside from the actual personality of the person, which can only be verified through close interaction through time, it is your perception of your potential partner that attracts you to him/her, whether the person of your affection truly has that kind of personality or not. This could probably account for a statement commonly heard from men and women on their failed relationships: "I thought he/she was this kind of person."

REFERENCES Alim, I., and K. Yuto. 2003. Mc21-A, a Bacteriocidal antibiotic produced by a new marine bacterium, Pseudoalteromonas phenotica sp. nov. O-Bc 30T, against Methicillin -Resistant Staphylococcus aureus. Antimicro. Agents Chemoth. 47: 480 - 488.

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

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

"Since my involvement with Wiley and environmental endocrinology, I have become more keenly aware of the nuances of hormonal interaction and understanding the molecular aspects of hormonal relationships," said Courtney Paige Ridley M.D."It answers the questions being posed regarding cancer and other dysfunction afflicting not only menopausal women but those women with significant alteration of cycle created by interaction with our estrogen toxic environment."

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

Is attraction a matter of chemistry? Maybe. According to scientists, the attraction between animals of the opposite sex is all about chemicals called pheromones. The effect of pheromones in behavior of insects is the most studied to date. It has been observed, at least in some experiments, that pheromones are responsible for communication among same species and colony of ants. The horrible odor released by skunks to ward off enemies is said to be a kind of pheromone. Some species of apes rub pheromone-containing urine on the feet of potential mates to attract them.

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.

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.

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.

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.

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.

- The women's ovulation period - The food that men eat - Perfumes and other scented body products - And lastly the use of contraceptive pills.

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.

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.

Probably not! There is wisdom in knowing yourself: who you are, your beliefs, values, and dreams. And do not pretend to be someone you are not. Fooling another person by making him/her think that you share the same values and beliefs is only going to cause you both disappointments. When you are healthy in all aspects, attractiveness becomes a consequence and not an end. As mentioned in the Klohnen and Luo's research, a person's sense of self-security matters, perhaps even beyond attraction. But remember: do these things for yourself and not for other people. Only then can you truly harness your attractiveness as a person.



It took great skill and will power to complete this article on homemade pheromones. We also request you to use your skill and will power to understand this matter.
Author Dave Thomson
 
 
 


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