Ways To Add More Romance To Your Love Life 

Everyone likes romance, don't they? When we think of a romantic date we conjure up images of moonlight, fine wine, flowers, chocolates, candlelight dinners and dancing close under the stars. Are there ways to help keep that romance and magic alive as a relationship progresses? Certainly there are. Let us take a look at ways to do just that.

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

Writing something about ingredients in pheromone perfume seemed to be something illogical in the beginning. However, with the progress of matter, it seemed logical. Matter just started pouring in, to give you this finished product.

Starch hydrolysis test, Protein hydrolysis test, Lipid hydrolysis test, Oxidative / Fermentative test, Salt concentration (0%, 3%, 5%, 7%, 10%), TCBS, Growth in Temperature, 42??C and 47??C

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

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.

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 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.).

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.

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.

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

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.

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].

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

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.

Scent is very romantic so wear some for her and she in turn should wear some for you. Everyone likes to smell great and smell works on the pheromones of our system to make us even more attractive to our significant other. Ladies spritz a touch of your favorite fragrance on your pillow so he smells your scent as he sleeps. Or write him a romantic note and spray a touch of your perfume on it and slip it into the pocket of his jacket of his briefcase. Talk about a touch of romance that will stay with him all day long until he sees you again.

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.

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.

Customarily, Tongkat Ali has been a botanical found throughout Southeast Asia where In Vietnam it is regarded a cure for everything. In the sex-crazy country of Thailand, it has been used by men as a virility tonic for centuries. And in Malaysia, it has been harvested heavily primarily to be sold in the United States (apart from being used at home).

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

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.

Writing something about ingredients in pheromone perfume seemed to be something illogical in the beginning. However, with the progress of matter, it seemed logical. Matter just started pouring in, to give you this finished product.

Icebreaker Pheromone Cologne

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 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.

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

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

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-)

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

Many different pheromones products are now available on the market. Pheromone products come in all shapes and sizes. For example, you have an option to buy pheromone colognes, pheromone oils, pheromone perfumes, or pheromone concentrates. Also, pheromone products are available in either unscented or scented versions. Of course, men and women use different types of pheromone products - but there are some unisex pheromone colognes, too. Androstenone and androsterone-based products attract women, while copulins attract men.

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.

Korber, D. R., J. R.Lawrence, H. M. Lappin-scott, and J. W. Costerton. 1995. Growth of microorganisms on surfaces. In. Microbial. Biofilm, Lapp in - Scott. Cambridge, UK: Cambridge university press.

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

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.

But Tongkat Ali, just like Yohimbe is a slow-growing shrub, and besides that, the active ingredient is found for the most part in the root. The root, which grows deep into poor hard soil, is very difficult to extract. Large holes must be dug to extract them and therefore in Malaysia, Thailand, and Vietnam, they have now started using the stems and branches of the shrub, instead of relying on the root. Just as there is nearly no yohimbine in the barks of pausinystalia species other than pausinystalia yohimbe, there are almost no active ingredients in tongkat ali stem and branches (the active ingredients are part of the root).

So, where to start? Use one product and pay attention to the reactions of men or women around you. Then try another pheromone product. The key with testing a product is to be persistent and to test in different situations. You cannot just apply pheromones then sit back and wait. Don't be discouraged if you don't see results immediately. Remember that everybody has unique body chemistry. Try different dosage or a different mix of two or more pheromone colognes. Testing is the best way to find out what works for you. And it's not expensive - there are a lot of free pheromone samples available - and pheromone kits for beginners as well.

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.

CONCLUSION: The present investigation was aimed to study the quorum sensing principle among seaweed epibiotic bacterial organisms. In the past few decades there was no findings of new novel antibacterial class compounds were identified. But, the pathogenic microorganisms show much higher rate of resistant development even to the potential antibiotics. So, there is an urgent need to discover new novel antibiotic compounds. The marine inhabitants such as microorganisms, seaweeds, invertebrates, etc., act as an undepleted source of wide range of natural products among which the seaweeds act as a potential source of antibiotic compounds. Currently the cross species induction / quorum sensing attracts the total attention of researchers in finding new novel drugs against multidrug resistant pathogenic microorganisms. So, the present study aim to find out the capability of seaweed epibiotic bacterial organisms to produce novel drugs against the animal and plant pathogens

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

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.

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-

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.

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.

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.

QUORUM SENSING EXPERIMENT NUMBER 1 In this present study, the producer and inducer strains were cross reacted to find out the production of antibiotic compound through quorum sensing. Totally three set of cultures were maintained as follows (along with one as control).

Do not just eat dinner together, instead turn it into a romantic dining experience. It has been said that it is "mood, not food that sets a romantic scene." You do not need to spend a lot of money for this one. Whether you are cooking a meal or ordering take out, put some fresh cut flowers in a vase, even if they come from your own yard, and then dim the lights, light some candles, pour some wine and enjoy a romantic meal together. Or maybe even cuddle up on the couch together with a pizza and watch one of your favorite romantic movies. Sharing a tub of ice cream is always a tasty way to be romantic together.

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.

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

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.

But on the northern tip of Sumatra (Aceh province of Indonesia) where the Tsunami occurred there still exist forests with a large areas of Tongkat Ali coverage. Tongkat ali from the Indonesia Aceh province can be purchased in wholesale and retail quantities from Sumatra Pasak Bumi.

This article was written with the intention of providing as much information on ingredients in pheromone perfume to its reader. Hope this objective has been fulfilled.
Author Dave Thomson

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