Based on some of your comments, and other inspirations, I posted a long page of links and excerpt/fragments covering visible bacteria sources and methods, that might be used for art or easier cheaper faster measurement of test results. Ver 2 july 12, 2016
Table of Contents
1. so the stuff glows under the 'black light' less before oxegen in depleted, then it glows more. Other stuff might glow under the black light so they do stuff like narrow laser and LED lighting and turning it off and seeing how many milliseconds the glow stays on, and non oxygen sensitive dyes can be used as a baseline. $120 of materiels to get into the game
2. $ 350, special broth makes color changing from 'indicator' non pathenogenic ecoli, part of the poop detection industry.
2.5 ONPG broth agent 5g $87, additional broth and spliced bugs unknown.
3. Grow some marine bioluminescent bacteria. $372, hope their cousins cholera, flesh eating sepsis bacteria don’t get into the mix and into our bodies.
Blue is alkaline and Red is acidy using this product chemical of a
bacteria, sort of a litmus test, maybe all I care about is if any
color is produced equals growth.
$354 for the germ plus the broth,
5. ammonia eating bacteria for aquariums can cause white materiel, but I have not been able to get it to work sensibly.
6. Consumption of dyes by bacteria, so far have not gotten anything to work, many suggested bacteria are pathogens I don’t want to work with.
The above lays down the problem and the path to a solution which I hope to lazily accomplish with this hardware:
"Cell-in-a-Box® encapsulation differ from those produced by others. Our competitors use substances such as alginate (a seaweed derivative) or chitosan, whereas our capsules are made principally of cellulose, a bio-inert material in the human body.”
some backgrounder and nomenklatura...
n 2016, Glowee, a French company started selling bioluminescent lights, targeting shop fronts and municipal street signs as their main markets. France has a law that forbids retailers and offices from illuminating their windows between 1 and 7 in the morning in order to minimize energy consumption and pollution. Glowee hoped their product would get round this ban. They used bacteria called Aliivibrio fischeri which glow in the dark but the maximum lifetime of their product was three days.
excerpt: "We monitor the metabolism of aerobic bacteria by using an oxygen sensitive fluorophore, ruthenium tris (2,2’-diprydl) dichloride hexahydrate (RTDP) that allows us to monitor the dissolved oxygen concentration in the nanowells. Using E.Coli K12 as a model pathogen, we demonstrate that the detection time of E.coli can be as fast as 35-60 minutes
device with smallest sized wells (1nL volume), had the largest
increase in fluorescent intensity. In comparison, the device with
100nL wells showed a slower rate of increase and those with 0.1mL
wells showed only a miniscule increase over time.
(how about two pieces of glass stuck together? plain? 1 sanded? laser or water jet etched? or a clutter slurry of minscule glass beads or something)
The powder of RTDP (ruthenium tris (2,2’-dipyridyl) dichloride hexahydrate) was obtained from Sigma-Aldrich (#224758). Liquid microbial growth medium, Luria Broth (LB) purchased from Sigma-Aldrich (#L2542)."
$30 for 250mg
"offline image analysis of the fluorescent intensity within the wells was calculated by using ImageJ (NIH, rsb.info.nih.gov/ij).
E.coli strain (OP50 and K12) was used for this study. Cells were grown aerobically at 37 °C in LB medium (10 g tryptone, 10 g NaCl, 5 g Yeast extract per liter). Bacterial cells in stationary phase were harvested after overnight incubation."
https://www.atcc.org/Products/All/10798.aspx (K-12) $60
Ruthenium based complexes have been widely used for detection and quantification of oxygen. Fluorescence is strongly quenched by oxygen. Its benefits are:
Long lifetime and emission around 613 nm minimizing autofluorescence background
Wide range of published applications
Ru(dpp)3(PF6) proved to be very useful for use in polystyrene membranes or beads
lifetimes of several microseconds, in this general review, the
outlook for measurement is looking not very plain
oxygen insensitive reference fluorophore, Nile blue chloride (222550, Sigma Samples were excited with a 488 nm laser for Alexa Fluor 488 and Ru(Ph2phen3)Cl2 detection and with a 635 nm laser for detection of the Nile Blue chloride. OSBs incorporating an oxygen sensitive luminophore (Ru(Ph2- phen3)Cl2) and two reference fluorophores (Nile Blue chloride and Alexa Fluor 488) were produced and characterized (Fig. 1A). The OSBs showed in their fluorescence spectra peak values near 505 nm (Alexa Fluor 488) and 610 nm (ruthenium complex) for excitation with a 488 nm laser, and near 655 nm (Nile Blue) for excitation with a 635 nm laser (Fig. 1B), which indicates the incorporation of all fluorescent molecules by the bead. The selected ruthenium complex has a long unquenched lifetime after excitation . Presence of molecular oxygen quenches the fluorescent signal in a diffusion-controlled manner (i.e. collisional quenching) and is described by the SterneVolmer theory . Fluorescence intensity is therefore inversely proportional to the quencher concentration
intra tissue/cellular 02 measurment
The application of chromogenic media in clinical microbiology. free review, much development is for pathogen detections that we would want to avoid.
a review which names names of manufacturers, tables
appears to be official us source, Tel: +1-973-564-7555,
ecoli for general water quality indicator detection may allow the use of non pathogenic bacteria EC166 or EC168, "quality control strains ATCC 25922 and 51446"
$250 for freeze dried mix to 5liters, includes growth medium function with color
http://www.atcc.org/Products/All/25922.aspx freeze dried biosafety 1, $60
http://www.atcc.org/Products/All/51446.aspx freezd dried biosafety 2, $225
poop pathogen detections
Our leader suggested this path of chromogenic detection to me.
"One option may be to use ONPG
Its a sugar that turns blue when broken down by a certain enzyme,
B-galactosidase. I can easily make an Ecoli strain carrying the
B-galactosidase gene for you. Whether or not this would be a
enough test for cell viability, I don't know.
based on recent post by djwrister, a look at implementing bioluminesence
Sodium chloride per liter
$60 26°C 78F
$354 26°C 78F
similar conditons, stronger light?
?only anerobic light emitter? same sigma aldritch broth
a different suggested medium? other than sigma aldritch type
Max Temperature: 18.0°C
damselae: Humans can be infected with the pathogen by eating infected
fish or swimming in brackish water in which the photobacterium can
travel up the urinary tract of a person. Humans can experience organ
failure, necrotizing fasciitis, and even death. Humans can survive
between 24-72 hours with the pathogen. Antibiotics, chemotherapy, and
radiation have been used to attempt to treat the infection, but have
failed. The best recommendation is to amputate the infected area of
the body before the pathogen spreads.
For safety, teachers might use only bioluminescent species which require 30 grams of NaCl per liter of medium. Such bacteria should not be able to grow in any animal. Another safety precaution is to use bacteria which can't grow at human body temperature. Many marine and soil bacteria can't grow at such temperatures.
"The bacteria I was growing is called Streptomyces Coelicolor M145 , A(3)2. it expresses an antibiotic called actinorhodin that is blue in a basic environment and red in an acidic environment."
Based on Lola's information I have collected some references:
ATCC medium: 1877 ISP Medium 1
Tryptone (BD 211705).........5.0 g
Yeast extract................3.0 g
Agar (if necessary).........15.0 g
Distilled water..............1.0 L
medium for final pH 7.0 - 7.2. Autoclave at 121C for 15
ATCC medium: 196 Yeast malt extract agar
Prepare Yeast Malt Extract Aqar (ISP Medium 2) (BD 277010) per
? no pigment production above 30 to 32C?
which type makes best colors???
Bacterial bloom triggered by high levels of ammonia. Nitrifying bacteria that consume ammonia reproduce rapidly to the point that they are visible, creating what looks like clouds of swirling, white smoke. Adding too many fish at one time, overfeeding, over cleaning with chlorinated water, and the use of antibiotic medications are common triggers. If bacterial blooms occur without these triggers, it may indicate inadequate nitrifying bacteria or biological filtration.
"The ammonia uptake in NOII is ≥15 mg N-NH3/hr/kg at 25°C."
For SALT & FRESH WATER
Nitrosomonas, Nitrospira and Nitrobactor
So far in my tests I got rapid creation of white particles when the "Nite out" was mixed full strength with full strength Smart n Final ammonia, I could not get residue at the very low concentrations suggested by the aquarium shop. Maybe the bacteria clumped and turned white while they were trying not to die from the ammonia concentration.
how about beet juice? and single strain probiotics?
so far the multistrain probiotics did not cause decolorization in my tests, but the unadulterated controls did decolonize... hmm,
cationic triphenylmethane dye, crystal violet,
"He found that most Gram-positive bacteria were sensitive to the dye, while most Gram-negative bacteria were not, and observed that the dye tended to act as a bacteriostatic agent rather than a bactericide.it can cause tattooing.
Streptomyces fulvissimus CKS 7
streptomycetes are Gram-positive, contradicts that this species eats the staining dye?
seems to produce an antibiotic that is cousin of the toxin of b cereus.
not usually pathogenic
90% within 10 h under aerobic culture condition
Crystal Violet could be used as sole carbon source and energy source for cell growth
Direct Blue 151 (DB 151) and Direct Red 31 (DR 31)
cereus showed maximum decolorization of 93.37% for mixed dyes under
optimum conditions. This was followed by B. cytotoxicus, Bacillus sp.
L10, and B. flexus showing 92.77%, 86.86%, and 85% decolorization,
respectively. A study conducted by Ponraj et al.37 showed that
Bacillus sp. has high decolorizing capacity for Orange 3R. They
isolated Bacillus sp., Klebsiella sp., Salmonella sp., and
Pseudomonas sp. from textile effluent and analyzed the decolorization
of Orange 3R under optimum conditions and reported that Bacillus sp.
and Pseudomonas sp.
B. cereus, which causes a foodborne illness similar to that of Staphylococcus. Although most species of Bacillus are harmless saprophytes, two species are considered medically significant: B.anthracis and B. cereus;
B. cereus, cheapest 60 dollars
Pseudomonas aeruginosa strain ZM130
reactive red-120, reactive black-5, reactive yellow-2, and reactive orange-16) in the presence of a mixture of four different heavy metals (Cr, Zn, Pb, Cd)
available dried, 290$ several versions level 1
virulence and pathogenticity databases