Understanding Vibrant: Algaefix, Polixetonium Chloride / Busan 77

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taricha

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The purpose of this post is to look closely at Vibrant to compare its properties to a known algaecide, Algaefix by API. This is intended to shed light on the active ingredients in Vibrant, explain its mechanism of action, and give people insight into its use and what to expect. It is not a recommendation to use or not use either product. (apologies, I'm not a chemist and I hope inaccuracies in my descriptions aren't too distracting.)

Background on Algaefix / polixetonium chloride
Algaefix by API contains 4.5% “dimethyliminoethylene dichloride, ethoxylate” CAS 31512-74-0 per MSDS and EPA documentation. This amount is the same in the various Algaefix Marine, Pond, etc labels. This chemical is a polymeric quaternary ammonium compound. Other quaternary ammonium compounds or “quats” (not polymers), such as benzalkonium chloride are in widespread use as sanitizers, antibacterial soaps etc. The Algaefix chemical is also known by the trade name Busan 77 or the more recently EPA-favored name polixetonium chloride, I will refer to it by these or simply “the polyquat.”
Polixetonium chloride is a well-known algaecide, registered in the U.S. since 1971. The EPA documentation on it is thorough and has a number of useful bits of info. This 65 page EPA draft risk assessment from 2020 covers most relevant info, including the material below. This chart shows the concentrations at which it was found to have an inhibitory effect on various “algae”.

NwSAslRbmC9iOhq-EFOn_x4tGxmEM2vSMN5nP2FdUdprL_9JoKoyPxQHEch-w4b0jg2hLRwJd6yf0PjMNYXha5Hd7SIVifh-EB-cYWKTELn6ymJKRsVcVQ9FKAR-II7s7I-dIHkb

904MYe40dlobzLdhoec8IyOQugFE7IHKBSdk8ZgcNxDXLrkXsTA__nOKU4QVYlgQ9f9rxQLc6Vj7CFlmadRBpWHUm3HrW5imZef6FeJeK2_m_8Ez8ZzOpMu2iEFSI9iAuJxUTI_w


The approximate concentrations for each group are: diatoms and cyano at ~0.1ppm, green algae at ~0.01ppm, and vascular plants at ~1ppm. The label dose of Algaefix results in an addition of ~1ppm per every 3 days. Perhaps unexpectedly, it does not seem to leave the system (though it would be expected to dissipate from the water).
“Polixetonium chloride is miscible in water [study #’s…] and is not expected to degrade by either abiotic processes [...] or biotic processes [...] Sorption to soil, sediment, and sludge is expected to be the primary route of dissipation, based on the fact that polixetonium chloride is a quaternary ammonium compound that has a positive electrical charge.
A ready biodegradability study […] demonstrated stability to microbial degradation in a WWTP [wastewater treatment plant], which is consistent with the results of other non-WWTP microbial degradation. Therefore, for polixetonium chloride, there is no apparent route of chemical or microbial degradation.”

One final note on known properties and uses of polixetonium chloride is that this same chemical(nih.gov) under the name Bubond 60 is also used as a coagulant / flocculant. This helps shed light on reports of increased water clarity with aquarium usage of the product.

A paper shared by @jeffww illustrated the use of a distinctive Bromphenol Blue color response (non-pH) to detect quats dried on surfaces. I noted a quat-like color change with both Algaefix and Vibrant, but in none of almost a dozen bottled bacteria hobby products or saltwater. After that, a more thorough investigation of Vibrant and Algaefix properties was undertaken. Below is the first part of those results.


Part 1: Professional Lab Tests indicate indistinguishable principle ingredients in Vibrant and Algaefix - polixetonium chloride
Samples of Algaefix and Vibrant were poured from the product bottles into labeled 50mL centrifuge tubes and shipped to a lab for NMR testing. 1mL of each was vacuum-dried and the residue taken up in heavy water for analysis by 13C and 1H NMR. A separate partially-used bottle of Vibrant was sent to @jeffww for FTIR comparison to Algaefix in another lab. Those samples were also dried under vacuum for FTIR analysis.

A) 13C NMR
This is a test that generates a magnetic resonance response from Carbon atoms in the sample. The resonance refers to the frequency at which certain carbon atoms will oscillate when pushed around by magnetic fields. Their preferred frequency depends on their local environment - that is, what they are bonded to. Thus a 13C NMR is a probe of the chemical structure of the compounds, with each Carbon atom in the compound being shifted to a different place in the graph by what it is bonded to. More thorough intro for 13C NMR in this text, and a follow-up.
C16dHaUioEVk0GbjL5HbuqAtq1uZg1CAwdd0PDtFVcsgfheBBnMFEIBuDVV6IIYG8wI2C6D9-QK8sFrErsa8Ki8wAbjgjKLyvu_ki-jtHxtjSFgf_XqW4OohDh0qvyGvjIZqpz28

In the above graph you can see that the samples of Algaefix and Vibrant possess only Carbon in the same bond environments as each other, and the Carbon atoms are in those environments in the same ratios.

A wider view shows the blank around 170 ppm chemical shift where aspartic acid (and any other amino acid) would have a peak, if there were any in the samples.
gBzbrg2n4HHqpKJZUBpI_PrW4zz85EAXTBTXYEmzV_-2BJ_Py5y8Tecbxg7BfZTXy_pnp3fI7DUjfH4FRKp95LwGC9uow1kjS-aNZN8jXok96ElC8pUAaaEj2P2_Ff3aA5Q4u9fZ



B) 1H NMR
This is the same process described above but for Hydrogen instead of Carbon. In the below data you can see that the hydrogen atoms in each sample also exist in the same bond environments and are in those particular environments in the same ratios.
fp6HDDZVlnqweOtJUOIzUQpTmjsKfS6stsoTfbZ-_EGtrh9LPU1-invwpYguiHKmaVdLjSUEswfkfhrYdmQ_nfouf3Hs98S1_9tgZg_PT006Yjf7YBS6_h7ZWxxnHG1OJOV6-pfq


Since both the Carbon and Hydrogen atoms are bonded in the same way in these vacuum-dried samples of AlgaeFix and Vibrant, this covers all possible organics in the residue.

C) FTIR
This process generates an absorption spectrum in the infrared region where chemical compounds are distinctive in what wavelengths they absorb.
uHsH1WBMlJgau3lOIZOhzgtk1yaehUtGXAxXjjz23thQ-UgBRuF0GftG8P7R3mSvZdLPlxLILlypWxGA0m9iJov3GTGMLFUVr2izE9eYyDLlbhdggOQg84JEizgRjY7R3oPZ-IKD

Again, the precise alignment of the absorption peaks and the consistency of their peak height ratios between the vacuum-dried samples of Algaefix and Vibrant tells us that the residues are the same compounds with no additions. (The addition of aspartic or other amino acids would be very obvious in this data as well).
In addition to being able to say that Algaefix and Vibrant contain only the same compounds in their dried residues, we can further compare that to a given FTIR spectrum for polixetonium chloride “Busan 77” (posted online).
yz9cA8UWj_-VzdeBJne4R_OWNFmCF9JJaBK2heOYlGg_RS9QYzxDSCdPzSEy7uNLSpHuoOBZB6xDHxNPmp1QblO8l5FlA10a88uRqfv6W06eAb7v7oTUYl50FcoEM3LoHrdh-90O

This level of agreement confirms further that what was measured by the above tests in samples of Vibrant and Algaefix is indeed the polyquat that is the well-documented label ingredient in Algaefix, polixetonium chloride.

next...
Part 2: quantification / comparison of the amount of polyquat in Vibrant and Algaefix

edit: 2/28/22
Part 2: post 16 here
Part 3: post 165 here

UWC response 2/28

edit: 3/10/22 & 3/15/22
Sample NMR test replication was done by @jda (so different hobbyist, different bottle from a different source, sent to a different lab with different technicians.) Results (1H NMR) he received are in post 731 here and (13C NMR) in post 778 here.
Overlays of the data I received and what jda received are below.
1H NMR
Overlay comparison.png

(same peaks shifted by the calibration difference between instruments)


and 13C
13C nmr overlay.png
 
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Dennis Cartier

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Wow, impressive work. I liked your first thread where you started looking into the possibility that Vibrant contains the same active chemical as Algaefix. The new methodology makes it even harder impossible to believe UWC's narrative that they 'force' bacteria into making an algaecide.
 

Joe31415

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You can't find Algaefix anywhere in Europe. I'm guessing it contains a banned substance.
Have you looked online? I found it on amazon dot co dot uk, ubuy dot co dot uk and a number of other *.it, *.pt, *.uk sites. Though I didn't look into them so I have no idea if they're reputable sites or what would happen if I actually tried to make a purchase (ie maybe it would stop me if I enter a European shipping address).
If you have a pond supply store, you might check with them. IIRC, that's what it's actually marketed for.
 

Ardeus

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Have you looked online? I found it on amazon dot co dot uk, ubuy dot co dot uk and a number of other *.it, *.pt, *.uk sites. Though I didn't look into them so I have no idea if they're reputable sites or what would happen if I actually tried to make a purchase (ie maybe it would stop me if I enter a European shipping address).
If you have a pond supply store, you might check with them. IIRC, that's what it's actually marketed for.

Those sites send products from outside Europe even though they have .pt .it etc extensions.
 

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You can't find Algaefix anywhere in Europe. I'm guessing it contains a banned substance.

You can find Vibrant though.
I wonder what that "banned substance" could possibly be... Hmmm. If you like the stuff, maybe you should stock up on it.

Based on what Jonathan just posted about the stability of the quat, it should have a nice long shelf life.
 
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Randy Holmes-Farley

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Thanks for posting this, taricha. I'll have to dig into it a bit later when I have more time to concentrate. :)
 
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taricha

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Part 2: The amount of active ingredient in Vibrant is the same as in Algaefix
Part 1 presented results showing that the active ingredient in Vibrant is indistinguishable from that in Algaefix, polixetonium chloride - even using professional lab tests that can probe the detailed chemical structure. This is a polyquat, a strongly cationic polymer, and multiple tests were done to quantify and compare the amount of this compound between the products. These tests revolve around the charge nature of these strongly cationic compounds, which is the chemical property that gives these substances their algae-killing effect. For these comparisons, I used 5 products: Vibrant, Algaefix (4.5% polixetonium chloride), Algae Control by Tetra (5.4%), Algae Clean Out by Fritz (5.4%), and Vibrant Freshwater. These tests show the sensitivity of the methods to clearly distinguish between the 4.5% concentration product and the 5.4% ones (20% more active ingredient), and allow the amount of active ingredient in Vibrant to be tightly established.

A. LaMotte Polyquat titration kit
This kit uses Toluidine Blue O as an indicator, and contains correction additives to guard against pH and hardness interference. It gives a nice sharp endpoint (example below) that can be easily compared to the target color card.
xxjDARs3KOD4QG7WWh5Ig2TYaSIwKHzw0mbzOZQO0n1E3zPJ2T8ynL-qx_AguJrXv1IaqVT15mck0mDSGQFy-yOkBra82x9Zo2jFwIv64LW2g1vUyRrRvHgX6h8NxNFuXsH3_XqU


Using this test on the 5 products listed generated the following data
zKxqAAC-6NeedVeOQRlLd_A4QDBA5PWVYxGoVSooCo0moY39Y7_ul4zb1QGwS8FeYX5X3h-VXr9VPHH2LW3IZ7k1J5lfg3u1OlMTsMHKXry95LSlJkplWN17tQ4XW5FBbpm62h9B

This data clearly shows that the amount of polyquat in Algaefix (4.5%), Vibrant, and Vibrant freshwater is indistinguishable, while the amount in Algae Control (5.4%) and Algae Clean Out (5.4%) is clearly higher. The slope difference between the two groups (average) is 16% higher which is in good agreement with the Algae Control / Algae Clean Out labels indicating 20% higher concentration.

B. LaMotte Quat titration kit (indicator)
Before using the polyquat kit, I worked with the LaMotte QAC kit for general quaternary ammonium compounds (QACs), which was not ideal. The color change was small and the titration step was interfered with by the coagulation properties of Vibrant and Algaefix, causing the color indicator to clump and fall out of solution during titration. I worked around the shortcomings to find a concentration range where the color change from the indicator gave a linear response, so concentrations between products could be compared. The color change indicator is a mix of bromophenol blue and methyl orange in their high pH form - and they each move in a way opposite their pH response when the cationic compound is added.
XKZqRPps6PcNwVD2Y2Sg2Ccvz5OJ8MFcb41YlqyltUykPvHQIMsq7xats7qXMZO3_6tTQCWtcl5laHxfixHmOcFDShHzCalLMHXGAMu2M3wg3A_iEaNsmP091XSJZO7u8_4zSG_a

Left is the pH shift behavior of the combined indicator and right is the response to the polyquat in Vibrant & Algaefix. This response was quantified by plotting the absorbance decrease at the peak of each indicator - bromophenol blue at 591nm, and methyl orange at 464nm - in response to increasing product concentration.
Below is the data generated by this analysis.
RyCA4gMZATms1xhZd5s8K1RHXJp4eBFsSxNXCqvpsIYIYmeAHPgd_v0UUGSFZuPSKMJSzB45ufUR9lbGpLVVjqPuoby-0yJnfybZlWlZnmlvt4GvvXfUkm7nzDvifefgReByxCsZ

Blue is Algaefix and red is Vibrant, the stars track the absorbance at 464nm (methyl orange) and the X’s track the absorbance at 591nm (bromophenol blue). Both indicators react identically in a concentration-dependent way to the addition of Algaefix and Vibrant.
Below is the same, but with 4 products.
0-I7dgoVhsbpXhYPSGBndMNKWzmWtB2phhqqubrkfKE4KwLzyNlR62Ogw_lCSgWXHukNNvH_N-CACLy59qU3Csbnhbbv3zfqbLZc1-8layZhxgiV71_UNFCk1JEIZw4udp4qqJ_H

Algefix is blue, Vibrant freshwater is red, and the higher concentration products Algae Control and Algae Clean Out are in yellow and green. This method can also clearly distinguish the 4.5% active ingredient product from the 5.4% ones (less product needed to generate the same indicator response). But again Algaefix, Vibrant, and Vibrant freshwater are indistinguishable in concentration.

C. Product reaction with sodium dodecyl sulfate (SDS)
When figuring out why the QAC titration kit coagulated and failed, I stumbled upon the observation that when the (cationic) polyquat is added to excess SDS (an anionic surfactant), a cloudy solution is formed by the reaction and the cloudiness is proportional to the amount of polyquat added.
t06fr10IeXYtdHC4di8mPTO0owzhidNdYIj8zl5pXDYh5BKfvC-77VUdriVbYOydLydQRri3Veehe-8qzKIBv9FkUEfLH64fXXsLdC4aaHmx4WYiG8KgJQFUc14VulpMhFxzsEPn


I used this fact to compare the concentrations of the polyquat in each product by the cloudiness / optical density generated upon addition of the products to 5mL of water with 0.250mL of 10% SDS. Below is the resulting data.
NL4iTuMrTO3XxlYvI0ltpGG7Tifvm1MhBlvvFBxlzTpG7X8E-pFQircHlcp7Dga-4lG1prclE42mZ5LDPYm_nryyb8WBWmS-KGk7J1RLKnCet1ktBtRlFK7IoNkWLM5KcQ8bsdLS

It is the same story as the other two methods of comparison. The 5.4% polixetonium chloride products - Algae Control and Algae Cleanout are easily distinguished as higher concentration than the 4.5% concentration product, Algaefix. Vibrant and Vibrant freshwater are on the other hand indistinguishable in concentration from Algaefix. The slope of the 5.4% products (average) comes out to be 19.8% higher than the other 3 products' average, as predicted by their labels.

So this leaves us with Part 1 demonstrating that the chemical in Vibrant is indistinguishable from the polixetonium chloride in Algaefix, and Part 2 demonstrating that the amount of this chemical is also indistinguishable between Algaefix and Vibrant, while products with even a 20% difference in concentration are easily distinguished.

There could be a part 3 with all the other dozen or so observations and measurements that show ways that bacterial products would be distinguishable from Algaefix, but do not distinguish Vibrant.
But it's of much less relevance. If it has the same algaecide in the same concentration as Algaefix, then the algae-killing behavior is entirely explained.

edit: here's Part 3, post 165
 
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Rick Mathew

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The purpose of this post is to look closely at Vibrant to compare its properties to a known algaecide, Algaefix by API. This is intended to shed light on the active ingredients in Vibrant, explain its mechanism of action, and give people insight into its use and what to expect. It is not a recommendation to use or not use either product. (apologies, I'm not a chemist and I hope inaccuracies in my descriptions aren't too distracting.)

Background on Algaefix / polixetonium chloride
Algaefix by API contains 4.5% “dimethyliminoethylene dichloride, ethoxylate” CAS 31512-74-0 per MSDS and EPA documentation. This amount is the same in the various Algaefix Marine, Pond, etc labels. This chemical is a polymeric quaternary ammonium compound. Other quaternary ammonium compounds or “quats” (not polymers), such as benzalkonium chloride are in widespread use as sanitizers, antibacterial soaps etc. The Algaefix chemical is also known by the trade name Busan 77 or the more recently EPA-favored name polixetonium chloride, I will refer to it by these or simply “the polyquat.”
Polixetonium chloride is a well-known algaecide, registered in the U.S. since 1971. The EPA documentation on it is thorough and has a number of useful bits of info. This 65 page EPA draft risk assessment from 2020 covers most relevant info, including the material below. This chart shows the concentrations at which it was found to have an inhibitory effect on various “algae”.

NwSAslRbmC9iOhq-EFOn_x4tGxmEM2vSMN5nP2FdUdprL_9JoKoyPxQHEch-w4b0jg2hLRwJd6yf0PjMNYXha5Hd7SIVifh-EB-cYWKTELn6ymJKRsVcVQ9FKAR-II7s7I-dIHkb

904MYe40dlobzLdhoec8IyOQugFE7IHKBSdk8ZgcNxDXLrkXsTA__nOKU4QVYlgQ9f9rxQLc6Vj7CFlmadRBpWHUm3HrW5imZef6FeJeK2_m_8Ez8ZzOpMu2iEFSI9iAuJxUTI_w


The approximate concentrations for each group are: diatoms and cyano at ~0.1ppm, green algae at ~0.01ppm, and vascular plants at ~1ppm. The label dose of Algaefix results in an addition of ~1ppm per every 3 days. Perhaps unexpectedly, it does not seem to leave the system (though it would be expected to dissipate from the water).
“Polixetonium chloride is miscible in water [study #’s…] and is not expected to degrade by either abiotic processes [...] or biotic processes [...] Sorption to soil, sediment, and sludge is expected to be the primary route of dissipation, based on the fact that polixetonium chloride is a quaternary ammonium compound that has a positive electrical charge.
A ready biodegradability study […] demonstrated stability to microbial degradation in a WWTP [wastewater treatment plant], which is consistent with the results of other non-WWTP microbial degradation. Therefore, for polixetonium chloride, there is no apparent route of chemical or microbial degradation.”

One final note on known properties and uses of polixetonium chloride is that this same chemical(nih.gov) under the name Bubond 60 is also used as a coagulant / flocculant. This helps shed light on reports of increased water clarity with aquarium usage of the product.

A paper shared by @jeffww illustrated the use of a distinctive Bromphenol Blue color response (non-pH) to detect quats dried on surfaces. I noted a quat-like color change with both Algaefix and Vibrant, but in none of almost a dozen bottled bacteria hobby products or saltwater. After that, a more thorough investigation of Vibrant and Algaefix properties was undertaken. Below is the first part of those results.


Part 1: Professional Lab Tests indicate indistinguishable principle ingredients in Vibrant and Algaefix - polixetonium chloride
Samples of Algaefix and Vibrant were poured from the product bottles into labeled 50mL centrifuge tubes and shipped to a lab for NMR testing. 1mL of each was vacuum-dried and the residue taken up in heavy water for analysis by 13C and 1H NMR. A separate partially-used bottle of Vibrant was sent to @jeffww for FTIR comparison to Algaefix in another lab. Those samples were also dried under vacuum for FTIR analysis.

A) 13C NMR
This is a test that generates a magnetic resonance response from Carbon atoms in the sample. The resonance refers to the frequency at which certain carbon atoms will oscillate when pushed around by magnetic fields. Their preferred frequency depends on their local environment - that is, what they are bonded to. Thus a 13C NMR is a probe of the chemical structure of the compounds, with each Carbon atom in the compound being shifted to a different place in the graph by what it is bonded to. More thorough intro for 13C NMR in this text, and a follow-up.
C16dHaUioEVk0GbjL5HbuqAtq1uZg1CAwdd0PDtFVcsgfheBBnMFEIBuDVV6IIYG8wI2C6D9-QK8sFrErsa8Ki8wAbjgjKLyvu_ki-jtHxtjSFgf_XqW4OohDh0qvyGvjIZqpz28

In the above graph you can see that the samples of Algaefix and Vibrant possess only Carbon in the same bond environments as each other, and the Carbon atoms are in those environments in the same ratios.

A wider view shows the blank around 170 ppm chemical shift where aspartic acid (and any other amino acid) would have a peak, if there were any in the samples.
gBzbrg2n4HHqpKJZUBpI_PrW4zz85EAXTBTXYEmzV_-2BJ_Py5y8Tecbxg7BfZTXy_pnp3fI7DUjfH4FRKp95LwGC9uow1kjS-aNZN8jXok96ElC8pUAaaEj2P2_Ff3aA5Q4u9fZ



B) 1H NMR
This is the same process described above but for Hydrogen instead of Carbon. In the below data you can see that the hydrogen atoms in each sample also exist in the same bond environments and are in those particular environments in the same ratios.
fp6HDDZVlnqweOtJUOIzUQpTmjsKfS6stsoTfbZ-_EGtrh9LPU1-invwpYguiHKmaVdLjSUEswfkfhrYdmQ_nfouf3Hs98S1_9tgZg_PT006Yjf7YBS6_h7ZWxxnHG1OJOV6-pfq


Since both the Carbon and Hydrogen atoms are bonded in the same way in these vacuum-dried samples of AlgaeFix and Vibrant, this covers all possible organics in the residue.

C) FTIR
This process generates an absorption spectrum in the infrared region where chemical compounds are distinctive in what wavelengths they absorb.
uHsH1WBMlJgau3lOIZOhzgtk1yaehUtGXAxXjjz23thQ-UgBRuF0GftG8P7R3mSvZdLPlxLILlypWxGA0m9iJov3GTGMLFUVr2izE9eYyDLlbhdggOQg84JEizgRjY7R3oPZ-IKD

Again, the precise alignment of the absorption peaks and the consistency of their peak height ratios between the vacuum-dried samples of Algaefix and Vibrant tells us that the residues are the same compounds with no additions. (The addition of aspartic or other amino acids would be very obvious in this data as well).
In addition to being able to say that Algaefix and Vibrant contain only the same compounds in their dried residues, we can further compare that to a given FTIR spectrum for polixetonium chloride “Busan 77” (posted online).
yz9cA8UWj_-VzdeBJne4R_OWNFmCF9JJaBK2heOYlGg_RS9QYzxDSCdPzSEy7uNLSpHuoOBZB6xDHxNPmp1QblO8l5FlA10a88uRqfv6W06eAb7v7oTUYl50FcoEM3LoHrdh-90O

This level of agreement confirms further that what was measured by the above tests in samples of Vibrant and Algaefix is indeed the polyquat that is the well-documented label ingredient in Algaefix, polixetonium chloride.

next...
Part 2: quantification / comparison of the amount of polyquat in Vibrant and Algaefix
This is some Epic work!...Well done! This is the kind of work that helps bring a much clearer understanding of what we are or are not putting into our tanks....really looking forward to part 2!
 
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ScottB

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I promise I won't bring out the duck analogy again. The one that goes like, if it walks like a duck, quacks like a duck, swims like a duck, looks like a duck, flies like a duck, tastes like a duck... it might be a pheasant on Spring Break. Won't do it.

Thanks again JB aka @taricha.
 
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