5/26/2018

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.


This article is under GNU FDL license and can be distributed without any previous authorization from the author. However the author's name and all the URLs (links) mentioned in the article and biography must be kept.

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




We were a bit tentative when embarking on this project on pheromone product. However, using the grit and determination we have, we have produced some fine reading material on pheromone product.

Aside from the studies conducted on the relationships between mother and child, there have been studies conducted regarding the effects of pheromones in humans when it comes to attracting the opposite sex. Although there has been much criticism regarding this matter and how it was not acceptable to some people that attraction between a male and female human being could be reduced to a simple interplay between the pheromones in humans, several scientists have made some discoveries in this area.

So how does attraction figure in relationships? You have probably heard that attraction is a prelude, or a factor towards a relationship. Most probably, at least in the beginning; but attraction alone cannot make a relationship work. It is that attraction that makes you notice a person from the opposite sex, but once you get to know the person more, attraction is just one consideration. Shared values, dreams, and passions become more significant in long-term relationships.

Like in the case of animals and insects where the young often pick up messages through the pheromones left by their mothers, infants display distinctive characteristics of recognition. Such recognition led scientists to believe that pheromones in humans also function much like the pheromones in other species although in a less pronounced manner.

As a result of this finding, perfume has become as much a science as it has an accessory. Aromatherapy studies the effects of natural botanical perfume ingredients, usually in the form of oils in their purest state. Its looks at the effects upon mood and general well being, and applies the knowledge by improving the perfume in the environment and offering a variety of aromatherapy services aimed at improving people`s lives. Aromacology applies this same approach to any available scent, from flowers to sand, and the findings are currently assisting perfume manufacturers worldwide.

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

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-)

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].

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

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-

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.

Holmstrom, C, and S. Kjelleberg. 1994. The effect of external biological factors on, settlement of marine invertebrates and new antifouling technologies. Biofouling. 8: 147-160.

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

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.

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.

Firstly, the effect of perfume on our brains is not necessarily recognised though the effects are certainly enjoyed. Each scent, whether natural or synthetic, emits molecules which enter the nasal passages and eventually reach the olfactory epithelium. This part of the nose has a vast amount of receptor neurons which are each covered in hairs called cilia. These cilia trap the scent and send a message to the brain which enables a person to recognise the perfume. The ability to smell immediately has a positive impact. This is important as some research has shown that people without a sense of smell have a less happy life than those who can smell; scents trigger emotions and affect our mood.

Microdictyon tenunis Chlorophyceae Chondrococcus hornemonii Chlorophyceae Enteromorpha intestinalis Chlorophyceae Caulerpa cupressoides Chlorophyceae

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

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

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.

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.

The test organisms Vibrio species were isolated from seaweed as epiphyles, biofilm, sediment and puffer fish by using TCBS medium (Hi media) The pathogenic bacteria were collected from clinical laboratories.

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.

Do pheromones in humans really work like the pheromones in insects and animals? A number of scientists would like to believe that they do. However, there are also many who vigorously object to such a conclusion because if we equate pheromones in human beings with that of other species, this could mean that like animals, humans could be made to falsely respond to certain synthetic forms of pheromones.

Cappuccino J. G., and N. Shermann. 1999. Microbiology, A Laboratory manual, Fourth edition, Addison Wesley, New York. Davies, D. G., M. R. Parsek, J. P. Pearson, B. H. Iglewski, J. W. Costerton, and E. P. Greenberg. 1998. The involvement of cell to cell signals in the development of a bacterial biofilm. Science. 280: 295-298.

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.

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.

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.

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

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.

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)

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.

We were a bit tentative when embarking on this project on pheromone product. However, using the grit and determination we have, we have produced some fine reading material on pheromone product.



Icebreaker Pheromone Cologne



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.

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

Ines, M. M., B. Karaigher, U. Cepon, and I. Mahne. 2003. Variability of the Quorum sensing system in natural isolates of Bacillus sp., Food technol. Biotechnol. 41: 23 - 28.

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.

Lately, scientists are looking into the existence of human pheromones and its role in mate selection. There are many conflicting views in the realm of biology, chemistry, genetics, and psychology. Most scientists would assert that these do not exist, or if they do, do not play a role in sexual attraction between a man and a woman. But new researches such as that conducted by Swiss researchers from the University of Bern led by Klaus Wedekind are slowly making these scientists rethink their stand. Their experiment involved women sniffing the cotton shirts of different men during their ovulation period. It was found out that women prefer the smell of men's shirts that were genetically different, but also shared similarities with the women's genes. This, like in the case of insects and other mammals, was to ensure better and healthier characteristics for their future children. But researchers also cautioned that preference for a male odor is affected by:

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.

Burgess, J. G., E. M. Jordan, M. Bregu, and A. Mearns-spragg. 1999. Microbial antagonism: a neglected avenue of natural products research. J. Biotechnology. 70: 27-32.

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

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.

The ingredients used in perfume may improve your concentration and enhance your mood. There is a long list of scents that are said to trigger this stimulating effect. Perfumes that contain ingredients such as ginger, sandalwood and rose are claimed by aromatherapists and aromacologists alike to have a stimulating effect on brain neurons. The increased stimulation aided by the use of perfume assists in many everyday tasks, from more focused studying to extra stamina and energy, balanced weight loss to an improved immune system.

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

The discovery of pheromones in 1959 led to several developments in the areas of relationships and behaviors. Pheromones were originally associated with the chemical substances that animals usually excrete to send a message to their own species. Further research on the topic widened the definition of the word pheromones to include other species such as human beings. In fact, pheromones in humans became one of the favorite subjects of researchers all over the world.

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

Bryers, J. D., and W. G. Characklis. 1982. Processes governing primary biofilm formation. Bioeng. 24: 2451-2476 Burgess, J. G., K. Boyd, E. Armstrong, T. Pisacane, and D. R. Adams. 2003. The development of a marine natural product-based antifouling paint. Biofouling. 19: 197 -205.

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.

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.



This article has practically covered all points on pheromone product. Do you feel the same thing upon reading this article?
Author Dave Thomson
 
 
 


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