Search the Community

Website with an easy to digest format of the concept: https://reverserespiration.com/ Reverse Respiration; The Quick-Guide Reverse Respiration is a new, aquatic plant cleaning method designed by multiple COOP forum members to remove pests and algae off of aquatic plants without inflicting any plant damage, and the only residue it leaves is water. In fact, Reverse Respiration paradoxically, appears to accelerate growth and plant health. There will be much more on this forthcoming in; “Reverse Respiration Growth Augmentation”. Concisely, how Reverse Respiration works is extremely simple: It places plants in a high pressure, pure-CO2 solution that asphyxiates the snails, worms, eggs, aerobic bacteria and insect larva but actually enriches the plants with an intense CO2 environment. This is done for 12 hours, in the dark. The plants are then placed in plain water (an O2 environment) for at least 30 minutes which suddenly changes the pressure which kills off any remaining eggs or anaerobic bacteria (if any). This last step also helps destroy any remaining algae. Since testing began on Reverse Respiration in January 2022, we have seen no surviving pests, most algae are killed and no plant damage has been observed and although it’s early, we’re now even seeing some growth acceleration. As Reverse Respiration is quite new, more field testing will naturally improve our data quality. We feel more field experience is needed but there’s ever mounting evidence from new users posts to suggest that Reverse Respiration is the least toxic yet most comprehensive plant disinfection technique available that does no damage to the plants and has no residue. Below are abbreviated instructions to perform Reverse Respiration. Please post your results on the main Reverse Respiration thread below as we are accruing data from all users to improve the quality of data of the entire experiment. If you wish to learn how and why Reverse Respiration works in great detail, you may read the results of the full, 6-month experiment and/or post your results on the thread here: Reverse Respiration Full Experiment Instructions for Performing Reverse Respiration What You’ll Need: Common Seltzer Water. Container with loose fitting lid. Dark area for 12 hours. You’ll need as much seltzer as is required to fully submerge the plants you are cleaning. It makes the plants quite buoyant such that you’ll need to weigh them down to keep them submerged. 1-Rinse the plants. Fully submerge the plants in freshly opened seltzer water. 2-Cover the top with a towel or other loose-fitting lid (should not be airtight) and place in the dark for 12 hours. 3-After 12 hours, remove the plants and soak in plain water, preferably aerated for 30 minutes or more. Usually, tank water is sufficiently oxygenated such that you may skip this step. 4-Rinse to remove remnants of pests or dead algae. 5-Place in the tank. You may also watch microscope videos of many of our tests via the links throughout the experiment and you may download all of the reference and citational materials we used in PDF form from a link at the end of the experiment. Please post your results and try to include pictures at the end of the full experiment thread as the more data we acquire, the more comprehensive Reverse Respiration can become. Note: We have recently observed that some sparkling waters may not have enough carbonation to perform Reverse Respiration. Please use only pure seltzer water or unflavored club soda as we have tested these extensively with to date, 100% success. Thank you for trying Reverse Respiration. * Introduction: A Forum Collaboration Sometime in early January 2022, we began an experiment to design and extensively test a new, chemical-free, aquatic plant cleaning method known as Reverse Respiration. We began with three unrelated technologies, two energy-based and one-gaseous-based as we were uncertain which, if any, would succeed or prove efficacious enough to be viable. As each of the nominated technologies was tested and analyzed, we eventually distilled the process down to the gaseous-based technology nominee, Reverse Respiration. We were greatly assisted throughout the experiment by many consultations with @OnlyGenusCaps and @Odd Duck such that it eventually became a collaborative effort involving four COOP forum members: @OnlyGenusCaps and @Odd Duck as consultants as well as @Guppysnail and myself operationally. @OnlyGenusCaps quite literally initiated the research. He encouraged and vigorously supported designing the experiment, literally from its embryonics. As we did not yet know which direction this might take, all three different technologies were fully developed and enjoyed a full, factorial experiment designed for each technique. These experiments for each were performed to verify the viability, or to reject a concept, and @OnlyGenusCaps was kind enough to review and advise on every one of them. Ultimately, two of the methods enjoyed partial success but suffered insufficient proofs and much of the performance was too mired in theory for us to assert claims with any degree of authority. However, for Reverse Respiration, we were able to fully develop proofs for each proposal and concept, all of which are available to anyone for download and review (see links at end). The initial concept was born in a discussion with @OnlyGenusCaps. He recognized the potential as I was unaware of the current plant cleaning methods and their limitations. @OnlyGenusCaps convinced me that the potential was great enough to warrant the research. As an expert in plant physiology, @OnlyGenusCaps has been an invaluable consultant throughout the testing. He analyzed our microscope images and videos and helped us identify and verify what we had recorded. His input, data and advice and continuous encouragement literally made this manifest. This experiment simply would not have been possible without his help. @Odd Duck, a veterinarian and authority in multiple related fields, reviewed and edited our documents and was a consultant for us as well. She graciously offered and examined our many microscope images and videos, and her observations led to redirecting our efforts to a pH theory that eventually led to proofs for the entirety of the process. Reverse Respiration might not have been fully understood without her help. She also was extremely kind to review and edit the entire lengthy document and made corrections and suggestions as well. It truly would have been a deeply inferior product without her expertise and input. Just one month into the experiment, it began to reveal its reach was beyond mere plant disinfection when an unusual post-cleaning growth acceleration was noted. And although the initial intent was only for plant disinfection, I was contacted by @Guppysnail as she was the first to observe that Reverse Respiration behaved as both an algaecide and potentially, a growth stimulant. I’m no expert in this area nor do I have enough experience with growing aquatic plants to speak with authority, so to that end, I found myself repeatedly consulting with this knowledgeable, veteran aquarist who would no doubt recognize and implement growing techniques that would escape my observation. @Guppysnail’s input became ever more invaluable and frequent, identifying growth patterns, their indications, species-specific conditions and behavior, as well as identifying pests. By then, the experiment expanded to 2 locations and 14 tanks such that we elected to make this experiment a collaboration. To that end, @Guppysnail and I partnered on this piece both operationally and as co-authors and her commentary is indicated throughout the article. Growth stimulation and observation, of course, take far more time for results than mere disinfection. As such, while @Guppysnail is a co-author of this article, you’ll see much more of her content in Part-2: Growth Augmentation in the coming months. Again, I truly do apologize for the length. I failed to find a direction forward that was more concise without neglecting proof of concept, and we didn’t want to present any concepts without a minimum of one proof; many have several proofs and even some precedents. At the end of the article, you may find all of the imagery, videos, university and laboratory studies in PDF form and the citations used during the entirety of the experiment. This content is free to download or redistribute and all the links are at the end of the article. I only hope the length does not dissuade anyone from reading it! But thank you once again for giving me a space to present results of my experiments and for allowing me to take so much of your time. What follows is the complete experiment: Reverse Respiration A Chemical-Free Technique for the Removal of Worms, Eggs, Diatoms and Algae from Aquatic Plants in Less than 24 Hours Without ANY Residue Reverse Respiration disinfects aquatic plants without chemicals or special equipment by simply alternating the CO2 levels in the water in cycles that favors plants but not pests. Plants require CO2 and O2 during their day and night cycles. Reverse Respiration works by denying oxygen to pests during the CO2 cycle of the plants. So, with the plants immersed in a pure CO2 solution, they respire normally but all of the pests are asphyxiated. Then the next cycle is oxygenated, denying CO2 to any anaerobes. In both cases, when the plants desire CO2, we exclude O2, and the aerobic pests perish. When the plants desire O2, we exclude CO2, and the anaerobe pests perish. Upon completion some 12 hours later, there are no pests, no eggs, no algae, and the only residue is water. The original experiment design had a time frame of two months. This included a month of promoting pest breeding in the infested plants under test. Additional time was required post-cleaning to observe for long term plant damage or growth, and to observe if any eggs remained fertile. To that end, the experiment was expanded to six months, fourteen tanks in two locations, in order to facilitate testing all of the nominated technologies. However, initial observations of post-disinfection growth stimulation first noted by @Guppysnail caused us to extend the process an additional six months and is presently ongoing. A followup article by @Guppysnail, “Part 2: Growth Augmentation,” will explore several technologies that one can easily apply to stimulate growth, improve survival rates, and accelerate ‘melt’ and regrowth. Categorically Speaking The method and time for expiration of all the snails and worms we saw were similar, but vastly different than the timing for algae and live eggs. To that end, we’ve grouped the parasites into two categories, based upon the time required for expiration: Dynamic Entities (snails, worms, all pests with motility) and Static Entities (non-motile, i.e., algae, diatoms, eggs). Dynamic Entities identified (there were innumerable unidentified!) in the tests: Ostracodes (crustaceans) Various Copepods (crustaceans) Rhabdocoela (flatworms) Various Nematodes (roundworms) Planaria (flatworms) Detritus worms Collembola, common name springtails (hexapods) Pond snails Bladder snails Ramshorn snails Multiple (black and clear) smaller snails Limpets (snails) Hydra (simple invertebrates) Aphids Insect larvae Static Entities included the plants used in the test: Cryptocoryne retrospiralis Cryptocoryne wendtii ‘Green’ Vallisneria Contortionist Anacharis (Egeria densa) Susswassertang Java Moss Hornwort Java Fern Marimo Ball Rotala rotundifolia Anubias Amazon Sword Hair BBA GSA Diatoms Snail Eggs Static Entities included algae and pest eggs under test Please note: Throughout this article, we’ll be taking some semantic license in categorizing for brevity and consistency. All plant cell content that is not chlorophyll-a will be referred to as non-chlorophyllic cell contents (all non-chlorophyllic enzymes, lipids, pigments and proteins, etc.). All references to gas spaces in plants, regardless of location, will be referred to as chymatous zones. Plant Damage The first casualty in plant cleaning is inevitably some plant damage and usually some mass loss. Comprehensive testing requires control batches (samples without changes imposed) that very closely mirror the batches under test. The “controls” are parameters where we have natural or original vs a sample under test. Alum Hydrogen Peroxide Vinegar Chlorine Bleach Reverse Respiration Untreated Our first control was a microscopic examination of the cells and content from an anacharis leaf treated with one of four popular aquatic plant cleaning methods: alum, hydrogen peroxide (H2O2), vinegar, and chlorine bleach, as well as Reverse Respiration. Below we have specified timing and quantities for each procedure for a more linear comparison. Alum: 1 tablespoon per gallon, 2-day treatment. Relatively gentle, alum did minor damage to chloroplasts, but alum’s extreme acidity caused substantial dissolution of non-chlorophyllic enzymes and proteins (usually brown or yellow in color). Vascular activity continued, that and being multicellular are why the plant largely recovered from the damage. Anacharis Cleaned with Alum Hydrogen Peroxide (H2O2): 1 part 3% H2O2 + 1 part water, 10-minute treatment. A mild antioxidant, H2O2 partially dissolved some non-chlorophyllic enzymes and proteins and partially oxidized (yellowing) the chloroplasts. Vascular activity continued, that and being multicellular are why the plant largely recovered from the damage. Anacharis Cleaned with Hydrogen Peroxide Vinegar: 1 part vinegar to 15 parts water, 5 minute treatment. Vinegar treatment resulted in minor non-chlorophyllic enzymes and proteins dissolution from extreme acidity but less damage to chloroplasts. Vascular activity continued, that and being multicellular are why the plant largely recovered from the damage. Anacharis Cleaned with Vinegar Chlorine Bleach: 1 part bleach to 19 parts water, 2 minute treatment (neutralized with dechlorinator). Virtually all non-chlorophyllic enzymes and proteins were removed and approximately half of the chloroplasts were killed, making this the most caustic of all treatments tested. Vascular activity continued, that and being multicellular are why the plant largely recovered from the damage. Anacharis Cleaned with Bleach Reverse Respiration: 12 hours in CO2 solution in the dark followed by 30 minutes plain, aerated water in the light. With Reverse Respiration, we experienced almost no enzyme loss. Chloroplasts were a brilliant green from the intense CO2 infusion and appeared nearly identical to the untreated plant. Non-chlorophyllic cell contents (usually brown to yellow in hue) were clearly visible and virtually unchanged from untreated plants. Vascular activity is dense and has rapid chloroplast activity, almost identical to an untreated plant. Anacharis Cleaned with Reverse Respiration and Anacharis Untreated There was little question that Reverse Respiration by far is the gentlest plant cleaning method we tested. Of all the techniques tested, within the limits of visual, microscopic analysis, it inflicted the least damage to the cell contents, proteins, enzymes and chloroplasts, and the only cleaning process that did not slow or reduce vascular activity. Tales from the CRYPTS by @Guppysnail Most aquatic plants are grown with their leaves ‘emerged’ in air vs submerged. This is much less expensive and enjoys a higher rate of success than when grown fully submerged. However, the leaves evolve differently in air, thicker and denser, as CO2 is much easier to access and more plentiful in air. When eventually submerged, they cannot perform metabolically and begin to rot…the infamous “crypt melt.” This is normal, and once the old growth rots and falls away, new growth will replace it over time. As they are thinner and have less mass than most plants, even water changes or other significant events can send a crypt or a val spiraling into a melt/regrowth cycle. The reuptake of the ‘melting’ plant’s rotted material as nutrients, known as remobilization, occurs, but this takes more energy and time than if the leaves are pruned. Pruning helps initiate the start of new growth, as there are no leaves for nutrients or respiration triggering new growth. The new leaves, of course, grow under water and are now adapted for an aquatic environment. They tend to be thinner and more exposed to the environment for improved access to nutrients and CO2 directly from the water but primarily due to the CO2 being not as plentiful under water. Reverse Respiration accelerates the melting cycle by chemically simulating the pruning process on dead and dying tissue. Once again, we are taking some semantic license, so I’ll be referring to this phenomenon as “pH Pruning”. pH Pruning is manifested as a form of accelerated dissolution of exposed, decaying (melting) cellular matter, accelerating the ‘melt,’ by exposure to the low pH of the CO2 solution. The CO2 solution dissolves the decaying matter at a much greater rate than normal decay. Reverse Respiration seems to accelerate both the decay of the dying organics and the replacement with new, healthy growth, although the latter may be due entirely to the former. There’s some suggestion that the dissolving of decaying material in concert with the intense CO2 charging of the chloroplasts may be a factor in the growth stimulation we initially observed (see “Val” images below). This would NOT occur if the tissue was healthy and intact, for there would be a multicellular and vascular buffer affording time before any form of dissolution occurs. The rapid dissolution of the decaying matter takes place because of the phenomena cited in the section on pH shifting (see “pH Shifting” below). What we found was a stark contrast between the way healthy plants reacted vs damaged ones. No healthy plants, leaves, stem or roots showed any discernable damage. Those that were previously ailing, however, melted or distinctly showed more damage over time once returned to the aquarium. Note these plants and plant parts were the ones that were already dying. Once leaves and stems are damaged or dying, they will not recover in any situation treated or not treated. Healthy growth is not affected, and new growth continues unaffected. Anacharis elodea densa In this picture the are several strands of Anacharis elodea densa. All were treated using Reverse Respiration. The leaves most heavily infested with BBA are the ones that were already dead. Most algae will take advantage of dying and dead leaves and the excess nutrients they release in the process of dying. Those not infested but unhealthy leaves are turning clear and will melt entirely. Left untreated those leaves would melt away on their own but over a longer period of time. Freshly planted Val leaves treated with Reverse Respiration are here beginning their classic’ melt’ cycle. New growth has already begun unusually early, well before the melt cycle is complete. Many of those unhealthy leaves still appeared healthy to the naked eye but were not. One common observation when purchasing aquatic plants grown emersed is that they appear greener and firmer than they would under water. This is because they are, but it is due to being grown in air (see above) and these will die off for new growth once submerged. Under a microscope we are able to see the vascular tissue of the plants in operation. The Xylem and Phloem of the vascular system carry nutrients and water to and from the roots and leaves. The vascular activity delaying cell saturation of the solution protects healthy plants during Reverse Respiration. · The visibly dead and dying have no chloroplast movement under a microscope. Untreated Melting Val · The ‘healthy to the naked eye but in the process of dying’ components show vastly reduced chloroplast movement or almost none at all under a microscope. Treated Melting Val · Healthy vascular activity is easily seen under the microscope in the undamaged leaves with chloroplast movement being rapid and dense. Untreated Val Many trials were done on healthy and unhealthy plants producing these same results. The dead, red BBA is eaten by aquarium inhabitants. Tanks that did not have inhabitants to eat it saw the killed BBA turn red, and eventually it turned white and disintegrated. Healthy Val Leaf Untreated Val Leaf beginning Melt Treated Healthy Val Leaf Treated Notice in the untreated, healthy leaf, the chloroplasts are widely distributed and easily discernable. In the untreated leaf with ‘melt’ beginning, you can see chaotic chloroplast distribution and clusters indicating low or no metabolic activity. The Reverse Respiration treated leaf looks nearly identical to the untreated, healthy leaf with highly discernable and plentiful chloroplast distribution, the only difference being an intense green from the saturation of the chloroplasts from the CO2 solution. To that end, our next control for this test was plant mass, that is: Does it lose any mass during the cleaning? We weighed the plants dry (when possible) to retain accuracy, or by using a fluid displacement method. Fluid displacement is more accurate than just weighing, since it ignores water weight on the plant; or you can weigh portions of the plant without cutting to determine if there was any measurable mass loss after cleaning. Unexpected Weight Gain With Reverse Respiration, we not only observed no plant damage, after treatment, we actually measured a GAIN in mass. What we found was the cleaned plant’s mass increased temporarily. This was not literal plant mass but actually the gas spaces within the plant, here, collectively referred to as the chymatous zones, filling with the CO2 solution. PEARLING (internal gasses escaping the chymatous zones) We found that an aquatic plant immersed in CO2 solution temporarily increases in mass, in concert with immediate “pearling.” This was further evidence of the chymatous zones filling with CO2 solution. Microscopic Image of Parenchyma The Chymatous Zones A variety of vacancies for gas exchange, respiration and buoyancy in plants are collectively known as the chymatous zones. Our intent was to demonstrate if these zones are indeed filling with our CO2 solution as an explanation for the mass gain. We needed to verify that the chymatous zones did indeed fill with the CO2 solution. Microscopic verification of this event was beyond our ken, so we elected to derive this mathematically. We used data from a now historic 1994 study done on chymatic pressure and area in leaves, done at the Department of Plant Biology, Louisiana State University. For this test specifically, we soaked a 671mG pothos leaf in CO2 solution for 12 hours. We calculated the chymatous zone area of this leaf sample and determined it could have enough vacancy to contain approximately 65uL of fluid if 100% of the internal gasses were displaced by water. In 12 hours, a CO2-soaked leaf gained about 67uG of mass, nearly equaling the weight of 65uL of water! The gas spaces were capable of containing 65uL of fluid which weighed 67uG. Yet within a 24-hour period, the plant had yielded back 98% of the weight gain, nearly returning to the original weight! This suggests liberal fluid passage into and exiting the plant, via the chymatous zones. Pressure Tactics Extensive testing of internal fluid and gas pressures in plants was performed at the Department of Plant Biology, Louisiana State University. In that 1994 study, they found that an aquatic plant’s internal CO2 pressure is about 3.5X greater than the CO2 pressure in the surrounding water. The CO2 solution, however, is much higher pressure than the normal atmospheric CO2 and that of the plant. The unopened container of the CO2 solution has some 17X more pressure than the natural atmospheric CO2 pressure but almost immediately stabilizes at about 3X the normal atmospheric CO2 pressure upon opening, where it remains on average for approximately 72 hours if undisturbed. Undisturbed CO2 solution retains enough CO2 to prevent any O2 saturation for approximately three days. Blue is plain, CO2 solution. The same test was made on Diet Pepsi (red) showing the additives in Pepsi inhibiting the rate at which it goes flat. A plant immersed in the CO2 solution begins to immediately fill the chymatous zones with roughly the same pressure CO2 as is throughout the solution, and within minutes, replaces all gasses from inside the plant with the CO2 solution. The CO2 solution is now both internal and external to the plant, effectively cancelling pressure impacting the plant. Pothos leaves deliberately hyper-stimulated with vibrational energy to simulate caustic conditions, performed in both plain water and in CO2 saturated water. The frontmost leaf was cleaned in plain water, and the leaf in the rear was cleaned in the CO2 solution. If the pressure is the same inside and outside of the plant, the plant experiences no pressure. We tested the pressure equalization theory by allowing the filling of the gas spaces in the leaves with the CO2 solution and then subjecting them to vibrational energy of normally fatal intensity. Notice the leaf cleaned in plain water (front) suffered significant damage, whereas the leaf cleaned in CO2 solution (rear) was nearly unscathed---further supporting the theory that having the same fluid inside and outside of the plant, the contiguity of masses---the energy is then the same throughout and around the plant, so relatively no energy impacts the plant. Snails, Worms, Algae and Diatoms Entities with more complexity than plants are less fortunate. Just 30 minutes in the CO2 solution eliminated 100% of the snails, worms and pests of all kinds, via asphyxiation. Worms of all types seem to expire almost immediately. Snails expired by minute 20. Unhatched eggs, algae and diatom expiration required 12 hours. Below: Images of parasites after being immersed in the CO2 solution for 30 minutes: Nematodes before CO2 soak After 30 minutes in a CO2 solution. Nematodes lost most of their mass and most of their elasticity. A Tough Egg to Crack Ostrich eggs Eggs proved to be another matter altogether. While the snails lost most of their color over a 30-minute treatment, (Snail Eggs-30 Minutes of Reverse Respiration) the embryos were still alive and in motion within the egg although their motion slowed considerably. Snail eggs after 30 minutes (left) and 12 hours (right) in CO2 water The snail egg on the left survived 30 minutes in CO2 solution but lost most of its coloration, and according to the study cited below, is likely sterile. (A study done in 2000 by the University of Miyazaki in Japan had demonstrated that nematode eggs subjected to 20% lower of oxygen levels than normal atmospheric levels for just 4 hours became unviable despite the still living embryo.) However, the snail egg on the right was identical but was killed by 12 hours of asphyxiation in CO2 water. Our intention was then to incubate the discolored eggs and watch for hatching. However, we found the 12-hour cycle of CO2 solution followed by just 30 minutes in aerated water (the “O2 cycle”) resulted in 100% expiration of all eggs and circumvented the need to do so. 12 hours of CO2 solution effected 100% egg elimination: Microscope images of treated snail eggs. Note the internal fluid exits the dissolving egg on the left, leaving visible ripple patterns as the fluid leaked over the slide. Similarly on the right, we saw the embryo begin to dissolve and leak well into the egg. Although algae were not a component in this test originally, when we observed Reverse Respiration’s tangential effects as an algaecide, we extended the tests to explore any parallelism amongst the egg and algae expiration, as they reacted quite similarly. Most forms of algae we care about are simple, single-celled organisms that respire similarly as most plants. Yet despite algae using CO2 for respiration, the CO2 solution killed all forms of algae we tested within hours: Black Beard, Green Spot, Green Dust, Staghorn, Hair and Diatoms. Green Spot Algae Hair Algae Green Dust Algae Black Beard Algae Staghorn Algae Originally, we suspected that the algae death observed was caused by a lack of respiration. That is, the CO2 solution denying a cycle for O2 respiration, followed by an “O2 solution” (aerated water) denying CO2 respiration, killing the algae. But algae death occurs far too quickly (<9 hours) for it to be respiratory… pH Shifting PH Shifting is a widely used process that uses pH extremes to extract the proteins and specific nutrients from plants and algae. Common sports drinks with algae proteins often use precipitated plant proteins made through this method. How Reverse Respiration kills algae and eggs is not unlike the same action that is commonly used to create high protein, algae-based nutrient drinks: pH Shifting. Normally the pH is raised greatly as chlorophylls, plant proteins and enzymes are more soluble in a higher pH. Once the plant proteins have dissolved into the solution, organic waste and other unwanted elements are filtered out. The pH is then quickly and drastically lowered to precipitate proteins and enzymes that are insoluble under acidic conditions (or pH is restored to neutral, and precipitates are removed electronically, known as ‘isoelectric precipitation’). This a generic explanation of pH Shifting as there are variations, but all operate in a similar fashion. Algae protein solubility per pH For our purposes, we employed a variation on this theme as precipitating the algae proteins is not for harvesting them but rather to act as an algaecide. Notice how the plant proteins begin to become insoluble as the pH falls, particularly so below a pH of 4.5. The CO2 solution’s low pH of 3 precipitates most of the proteins and enzymes in the algae cells, destroying (denaturing) them in the process. Most of the precipitated algae cell contents, however, dissolve in the CO2 solution's acidic conditions, forming extremely minute quantities of carbonate salts that simply rinse off. Plants have many proteins and enzymes, but chemically, all but one, chlorophyll-a, become unstable at a pH below 4. Although most plant enzymes are insoluble below a pH of about 4, chlorophyll-a remains viably active as low as pH 2! Chlorophyll-A Stability / pH Chlorophyll-A Light Absorbance Efficacy / pH BBA, GDA, GFA, GHA, GSA...R.I.P. by @Guppysnail While working on another project with plants soaking in the CO2 solution for an approximately a 12-hour period, something very unexpected was noted. It involved various plants covered in various algae. Green Hair algae (GHA), Green Fuzz algae (GFA), Green Spot algae (GSA), Black Beard algae (BBA), Staghorn algae, as well as some plants completely coated and suffocating from brown diatoms. At the end of the day, the unused plants were placed back in the aquarium. But WAIT...where are the plants covered in brown diatoms? There they were appearing completely diatom free and vibrant once again. A mystery is afoot! The other plants and algae were examined. The green fuzz/hair algae appeared lighter, some near white and opaque. Other algae appeared unaltered. · The plants were returned to the aquarium for the evening. · Almost every shrimp in the tank converged on all types of dead algae. · In the morning the brown diatom plants still had no trace of brown to the naked eye. · The green hair/fuzz was entirely white and opaque and had been mostly consumed by the shrimp or dissolved. · Plant leaves almost entirely covered with green spot algae were now partially cleaned of the GSA. · The real shock was the plant nearly covered in Black Beard algae (BBA) and staghorn. Every drop of the BBA and staghorn turned blazing red and was being consumed or dissolving. A second test of this resulted in the same alteration to the algae but was returned to a tank with no inhabitants that would consume the altered algae. Over the course of 72 hours, all types except GSA dissolved on their own. The GSA saw about half fall off. However, 100% of the GSA died and no more grew. In yet another example, here we have BBA treated with Reverse Respiration, but I had no animals in the tank that would consume it, so its action as an algaecide is on display: BBA Before and After Reverse Respiration Treatment Without any predators to consume the killed algae, the BBA turns white and slowly falls off. At this point the prior project was paused, and we investigated this avenue. We simply had to know why it kills all algae, but not plants. The best potential for explaining our observations proved to be with diatoms. Diatoms have a different ratio of content such that there’s more non-chlorophyllic (brown enzymatic) content than green chlorophyll as opposed to common algae, yet it contains both. This seemed ideal to test this theory as if it dissolved most of the brown content (non-chlorophyllic enzymes and proteins), but the green, chlorophyllic content remained, it strongly points towards pH shifting. We found that virtually all of the algae proteins and enzymes are removed by Reverse Respiration, leaving expired or hollowed algae cells or cells simply incapable of a metabolic process. Only chlorophyll-a remains, such that the algae and even diatoms still appear bright green or form green patches due to the intense CO2 saturation of the chlorophyll-a (see below). However, in our testing, 100% of the algae samples turned red or white and died, as they no longer had any metabolic activity. Diatoms after 12 hours in CO2 solution---all enzymes are gone, leaving hollow cell walls and chlorophyll-a (green) remnants. BBA before and after CO2 treatment Hair Algae before and after CO2 treatment Diatoms before and after CO2 treatment GSA before and after CO2 treatment In all instances, we found most of the cell’s content is largely gone (denatured), leaving large gaps in and between the cells: Healthy BBA - Denatured BBA Once again, the only enzymatic remnant we found was chlorophyll-a, hence the dark green (darker here as they are stained for the microscope) in all of the images. Chlorophyll-a remained because it is the only one stable in a pH of only 3, but chlorophyll-a alone is not enough for metabolic processes to continue, so the algae perish. Timing is Everything Our final tests were to verify theories as to why algae perish within hours, whereas live plants seem immune and may actually enjoy growth augmentation from Reverse Respiration. It’s all about time: Left: Normal chloroplast and vascular activity Right: 4 days Reverse Respiration no activity Most algae are open to water flows, immediately and directly accessing water-borne gasses and nutrients, with no vascular systems to delay the process. This and other factors result in algae metabolizing at 10X to 50X the rate of terrestrial plants. Plants have a vascular system, a pseudo-circulatory system that requires time to bring gas and nutrients into the plant and distribute them. So, although it may appear stagnant, algae grow and perform nearly all metabolic activity much faster than plants. Too Much of a Good Thing? “Why then, can one desire too much of a good thing?”- As You Like It, circa 1600 While the vascular system (and being multicellular) protects plants from damage, we needed to determine if the CO2 solution ever damages the plants, for if not, it conflicts with our theories on why it kills algae so effectively. To that end we subjected the same anacharis plant to repeated (and unnecessary) treatments until we observed damage and found that the same action that kills algae will eventually kill live plants if the treatment is extended some 8X longer than the required treatment time. (It required a 12-hour treatment be extended to 96 hours to finally incur plant damage.) It is this inherent delay in the distribution of gasses and nutrients that allows the plants enough time to see the pests and algae expire, yet it avoids any damage to the plants as evidenced in these microscope images below of an anacharis plant being treated with Reverse Respiration (extended from 12 to 96 hours to test how long a vascular plant will tolerate this low pH): Day 1 - Normal Vascular Activity-Day 2 - Normal Vascular Activity Day 3 - Vascular Activity; some enzymatic dissolution-Day 4 - No Vascular activity It took approximately 4 days before we witnessed any irreversible plant damage. The pests and algae are eliminated within 12 hours, but live plants endure up to 96 hours without damage. The Difference is Night & Day Our initial intention for performing these treatments in the dark was that in darkness, the plants under treatment will not respire any O2, further depriving the pests of oxygen, therefore accelerating the expiration of the aerobic entities like pests and eggs. We did somewhat unexpectedly observe that the rate of expiration of the algae accelerated in darkness substantially as well. The algae expired at more than TWICE the rate when treated in the dark. In fact, some 12-hour expiratory events shortened to as little as 30 minutes simply from being performed in the dark. “Nothing and Something Create Each Other” (From Chapter Two, Tao te Ching, circa 500BC) Although it takes countless forms, mass, light, heat, x-rays, all are ultimately just energy. Be it a rock or a bolt of lightning, both are the same stuff, a form of energy. What a mass or a form of energy is defined as is relative and depends on the observer. It is the scale that creates the reality. If we were at the level of an atom inside of the rock, we’d see countless energetic particles moving near the speed of light. If we pull back, it’s just a motionless, dead rock. What makes it a mass is not inherent but rather where we observe it from. But in truth, liquid, solid or gas, light or electricity, it’s just the same stuff as the rest of the universe: energy. Or said in a more relativistic fashion, light (photons) is the same thing as electricity (electrons). It’s only “light” because we can see it. That is, we the observers, “create” light. An original handwritten letter from Albert Einstein with the E=mc2 equation sold for $1.2 million dollars at RR Auction in 2021 The most famous equation in history centers around this core principle. The Photoelectric Effect was first discovered by Albert Einstein in 1905. Einstein won a Nobel Prize in Physics for The Photoelectric Effect yet ironically enough, his most famous and important work, The General Theory of Relativity was passed over by the Nobel committee. However, The Photoelectric Effect is an integral component of Relativity. The Photoelectric Effect Before Relativity, Einstein proposed that all energy regardless of type is the same. To illustrate this, he correctly predicted that when light strikes a metal, that metal will release electricity because energy is interchangeable. Light (photons) can infuse metal atoms with more energy than they can contain, forcing them to release electricity (electrons). He called this The Photoelectric Effect. This phenomenon ultimately became the technology in digital cameras, solar cells and most likely, in the screen that you’re reading this on. When we first observed that Reverse Respiration killed algae more quickly in the dark, we finally understood how Reverse Respiration acts as an algicide. We used Einstein’s Photoelectric Effect to demonstrate how Reverse Respiration kills algae. Reverse Respiration kills algae by removing electrons from metals in the algae. As you'll see below, the metal most affected by Reverse Respiration is magnesium however, it also destroys the potassium and calcium amongst other metals inherent to common alga's chemistry. The “Lattice Energies” indicate the ability of each metal listed to enter into a chemical reaction. The greater this energy, the more likely a chemical reaction with that metal will occur. Of some metals commonly found in algae, magnesium followed by calcium are the most likely to form new compounds. This reaction can be catalyzed by a chemical reaction (such as the CO2 solution), electricity or light. When we first observed that algae perished more quickly in darkness, we realized that light (photons) was interfering with this algicidal reaction. The reason this occurs has nothing to do with photosynthesis, however. This reaction is about 10,000X stronger than photosynthesis. Light degrades its algicidal power due to The Photoelectric Effect. In essence, if photons (light) are present, the energy required (electrons) to effect a chemical reaction may not all be taken from the algae but rather, from the light itself, sparing the algae to some degree. If Reverse Respiration is performed in total darkness, the only energy available for this reaction must come from the algae, killing it. A well-known study done (see the chart below) on the reaction of chlorophyll in various, acidic conditions (as well as many other plants cellular contents-testing performance in stagnant waters and other potentially acidic environments) was performed in 1977 at the Department of Vegetable Crops, at the University of California in Davis. It became well known because perhaps not inadvertently, they made the one of the first Quantum Biology demonstrations ever, four years before it became a viable theory, although that was not their goal. What they found was most cell content that was denatured or in the process of being denatured, did so at 2X-4X the rate in darkness than in light. Chlorophyll Destruction by the Bisulfite-Oxygen System - PMC (nih.gov) As stated above, our original intent to keep the process in darkness was to prevent the plants from photosynthesizing and therefore creating oxygen when we're trying to asphyxiate the pests. This observation added a secondary import to performing Reverse Respiration in darkness: That Reverse Respiration kills algae up to 40% more quickly in the dark. The CO2 solution removes and converts the algae magnesium to a magnesium bicarbonate salt by stripping it from the algae and killing it. Eventually once dried, the newly formed magnesium bicarbonate is unstable, it loses any remaining H2O and CO2 content and ultimately manifests as a white powder residue in the evaporate, magnesium oxide - a common antacid. The first image above are the distillates, distilled water at a neutral pH of 7 and the same water used to dissolve the evaporate from a Reverse Respiration session. The insoluble solids are filtered out to compare the chemistry of the solutions. The brown coloration is the cell contents stripped from the algae (as well as general dirt that the effervescence ‘scrubs’ off of the plants), now dissolved in the water. The white powder, the magnesium carbonate precipitant (it becomes magnesium bicarbonate in water), dissolves in the fresh water, raising the pH and alkalinity. On the immediate above left the rise in nitrate, nitrite and cyanuric acid are indicative of the brown, decaying organic matter from the algae and is relatively little. But the (magnesium) carbonate content is now quite literally, off the chart, as predicted by The Photoelectric Effect. On the above right is a standard flame test of the precipitant solution with a white-colored flame indicating the element is largely magnesium. Quantum Biology For a deeper peer into the subatomic governing this amazing reaction the entire study cited above is available for download in the PDF section at the end. If you have less time, although the famous quantum theorist Schrödinger initiated the idea of Quantum Biology with his findings, that relatively new science (<40 years) was largely developed by the British physicist J. McFadden. He was kind enough to make popular videos explaining this phenomenon. They are low budget productions, but the content is priceless: Professor Johnjoe McFadden's "Does Biology Need Quantum Mechanics?" at Imperial College London's 2014 The Solution is the Solution Admittedly, the path to this solution was much more convoluted and complex than we had imagined, in part because our initial intention was solely to design a ‘chemical-free pesticide’ to clean aquatic plants. But observations soon resulted in expanding the testing and analysis to explore these parallel events, as it soon evolved into a multi-faceted, pesticide – algaecide – and growth stimulant. Cleaning Aquatic Plants with Reverse Respiration Providing scientific proofs involved months of testing, hundreds of references, several consultants (several COOP forum members!) and reviewing dozens of laboratory studies. Yet as convoluted as designing the procedure and providing proofs was, the resultant process was distilled down to an extremely simple solution, so long as one observes proper timing and lighting. We found a literal and figurative solution by immersing the plants in carbonated soda or seltzer water for 12 hours in the dark followed by 30 minutes in aerated water, in the light. Plain, unflavored seltzer has the correct pH, pCO2 (pressure), anoxic state, and timing of decay to effect everything cited above in our testing. Aerated water is sufficient to serve as the O2-only portion of the cycle. Reverse Respiration Instructions: 1. Immerse the plants entirely below the surface in freshly opened seltzer water. It should be an open lid container such that the pressure does not build. You may need to add a weight to keep the plants submerged as the effervescence and high pressure makes them highly buoyant. 2. Place in the dark. Stir gently to help solution penetrate difficult areas but do not disturb once placed in the dark. 3. Remove after 9 hours-12 hours. Rinse. 4. Place in fresh water and aerate for a minimum of 30 minutes in the light. Rinse. For all of the reasons cited above, in less than 24 hours, there will be no worms, no larva, no eggs, no snails, no algae, and all of the carbonation and excess oxygen will have dissipated, …the only residue being WATER. IN MEMORIAM: Live snail embryo before and after 12 hours of Reverse Respiration Hair Algae before and after 12 hours of Reverse Respiration BBA algae before and after 12 hours of Reverse Respiration GSA algae before and after 12-hours of Reverse Respiration Diatoms before CO2 Diatoms after CO2, saturated green Close-up--CO2 saturated Plant damage (anacharis) after repeated, redundant Reverse Respiration treatments to test plant stress over a number of days: Day 1 (healthy) Day 2 (healthy) Day 3 (minor damage) Day 4 (dead). In summary, after six months of testing and treating dozens of plants, in fourteen test tanks, at two facilities, we had zero pest survivors, zero surviving eggs, and…zero plants perished. In fact, the opposite occurred… G R O W H What we observed in the subsequent days was beyond paradoxical. Our intent was to quantify how much plant damage occurs and if it is reversible. What occurred truly surprised us and extended the experiment to an estimated six additional months longer than previously intended. The first obvious change in the plants upon immersion in seltzer is they become rigid and deeply green. Even crypts and vals stood straighter and more erect. The relatively sudden environment of pure CO2 saturates the chlorophyll in the plants, making them a deep, intense green within hours. In addition, the seltzer replacing the gases in the chymatous zones add mass and rigidity to the plants. But it did still more. We measured the mass of a hornwort sample in the control tank vs. the same after the first cleaning session. Just days after the cleanings, we noticed that the hornwort that was cleaned appeared brighter, greener and slightly larger. In the days to come, the hornwort subjected to treatment became much larger than the original control batch. Inside of a month, the cleaned hornwort was some 80% larger than the control (untreated) hornwort! External energetic stimulation of plant tissue to accelerate growth is in common use. While it’s not fully understood, applying external energy to plants, seeds, and roots seems to have a similar effect on plants as light energy. The growth stimulation in both cases was short lived though. It seems to accelerate for several weeks, producing longer, taller, and thinner plants initially, which eventually linearizes to normal growth and appearance. We noticed the growth of the control hornwort ‘caught up’ with the treated hornwort after an additional 4-5 weeks. And while many studies have been done, one famous study done at the Tokyo Institute of Technology in 2014 cited some theoretical possibilities. This study, which was done on the effects of externally applied energy to radishes, had shown accelerated growth up to a staggering 87%-150%! The study noted that auxins, plant hormones that regulate growth, saw their production greatly stimulated after the treatments. In fact, they grew so rapidly that they distorted the shape and size of the plant, albeit temporarily. In a matter of weeks, growth normalizes. While no further testing was performed by us, this observation served as an important indicator that Reverse Respiration, unlike all other plant disinfection methods, inflicts little to no plant damage. ******* ADDENDA In this section we intend to post new findings from ourselves, other users or any meaningful contributions to the experiment. Posts in this section may be unproven or theoretical and are noted as such if so. Addendum-1: SODA POPS A study performed at the University of Austin in 2018 was done on champagne to determine oscillation rates of the bubbles. Some have rightly raised the difficulty of cleaning plants that are deeply netted or suffer difficult to penetrate areas; that is, the formation of ‘micro pockets’ of trapped air or even water, shielding the pests we’re trying to eliminate. This of course is an issue with all forms of treatment in a liquid that is static. One of the interesting byproducts of using a carbonated liquid as the cleaning agent is that the liquid is dynamic or in continuous motion. While trapped, protected areas must exist to some extent, the following could suggest that the survival of ‘micro pocket shelters’ in carbonated water is far less likely than static cleaning solutions. The tests performed for the study above at the University of Austin in 2018 were done on champagne but we performed our own and found very similar results with seltzer water. Seltzer effervesces at an amazingly predictable rate. In fact, it bubbles at many rates but at two rates primarily. These rates don’t vary much over time, but they do lose intensity over time. These rates are known as oscillations as seen in the oscilloscope image below: Seltzer water has two dominant oscillations, one being infrasonic at about 1-3 times a second (1Hz-3Hz) and another much higher at about 7000 times per second (7kHz). The low frequency, infrasonic vibration of the first oscillation literally shakes the plant during the entirety of the bath. Simultaneously, the approximately 7000Hz oscillation behaves not unlike a pseudo ultrasonic cleaner as it forms immeasurable amounts of microscopic bubbles which pop, sending vibrational energy throughout the solution. Our hypothesis is that this energy is strong enough to penetrate microscopic nooks and crannies or to burst standing air or water and dislodge most anything from the plants' surface with surprising power, known as ‘cavitation.' This is known as cavitation and is how commercial ultrasonic cleaner’s work. Although we did not test this, we saw no pests or eggs survive even after 6 months. Of course, the degree of any additional deep cleaning and penetration this affords is speculation at this point. However, it could suggest that a combination of the very high pressure of seltzer with the mechanical cleaning action of the seltzer bubbles cavitating should at least improve the likelihood of micro pocket penetration. As an example of the incredible pressure, here the 300:1 pressure of seltzer water applied to algae cells forces the contents almost completely out of Marimo algae cells at around second 45 in the video: Marimo Algae Drained by CO2 Pressure Here is a sample of the recording of the oscillations in seltzer. A hydrophone was immersed in seltzer to measure the frequencies of the oscillatory energy. If you listen closely, you can hear the very high frequency ‘whistle’ of the fast, microscopic cavitation bubbles popping en masse along with the low rumble (and occasional large bubbles crashing into the microphone!) of the much larger, effervescence bubbles: Recording of Seltzer Oscillations This addendum is speculative and has not been tested beyond what is presented here. ******* The Cinema Below are links (Ctrl + Click) to microscope videos we shot during the test period (10X-2500X, 40x magnification unless otherwise noted). They show various entities in the tests and provide a live, microscopic look into the eggs, how they protected the snail embryos, and how they eventually expired. Seventy-two microscope videos were shot for these tests, all of which are available for your viewing or to download here: Aquatic Animals and Eggs used in Reverse Respiration Tests under Microscope Aquatic Plants used in Reverse Respiration Tests under Microscope References PDFs of the Studies Below - Download Link Urban Forest Research Center, National Institute of Forest Science, Seoul, Agricultural and Life Sciences, The University of Tokyo, Yayoi 1‑1‑1, Bunkyo‑ku, Tokyo Judit Dobránszki (2021) Application of naturally occurring mechanical forces in in-vitro plant tissue culture and biotechnology, Plant Signaling & Behavior, 16:6, 1902656, PQM-1 controls hypoxic survival via regulation of lipid metabolism Thomas Heimbucher, Julian Hog, Piyush Gupta & Coleen T. Murphy Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA, USA Banerjee N, Hallem EA (2020). The role of carbon dioxide in nematode behavior and physiology. Parasitology 147, 841–854. Aerenchyma formation in the rice stem and its promotion by H2O2 Bianka Steffens, Thomas Geske and Margret Sauter Physiologie und Entwicklungsbiologie der Pflanzen, Botanisches Institut, Universita¨t Kiel, Germany Photosynthetic Response to Elevated Carbon Dioxide Concentrations in the aerenchyma of Typha Latifolia L. Leaves. John Van horne Constable 1993 Louisiana State University and Agricultural & Mechanical College Precision and Intelligence Laboratory, Tokyo Institute of Technology,Nagatsuta-cho Batangas State University, College of Engineering, Architecture and Fine Arts, Pablo Borbon Main II, Alangilan, Batangas City, Philippines Batangas State University, College of Engineering, Architecture and Fine Arts, Pablo Borbon Main II, Alangilan, Batangas City, Philippines Emerson Process Management Beverage (Carbonated Drink) - De-aeration J. Aquat. Plant Manage. 45: 76-83 In-vitro Investigations on Ultrasonic Control of Water Chestnut MEI-YIN WU1 AND J. WU Corresponding author: Center for Earth and Environmental Science, State University of New York, College at Plattsburgh Department of Physics, University of Vermont Aerenchyma Carbon Dioxide Can Be Assimilated in Typha latifolia L. Leaves John V. H. Constable and David J. Longstreth Plant Physiology Volume 52, Issues 1–2, September 1995, Pages 93-106 Aquatic Botany CO2 and O2 transport in the aerenchyma of Cyperus papyrus L. Michael B.Jones Broad oxygen tolerance in the nematode Caenorhabditis elegans W.A. Van Voorhies, S. Ward Author and article information J Exp Biol (2000) 203 (16): 2467–2478. https://doi.org/10.1242/jeb.203.16.2467-15 AUGUST 2000 On cryptobiosis and anoxibiosis: N. Dasgupta, A. M. Patel, B. A. Scott, C. M. Crowder, Hypoxic preconditioning requires the apoptosis protein CED-4 in C. elegans. Curr. Biol. 17, 1954-1959 (2007). [PubMed] Oxygen-Starved Worms JOHN F. FOLEY SCIENCE'S STKE • 27 Nov 2007 • Vol 2007, Issue 414 • p. tw432 • DOI: 10.1126/stke.4142007tw432 Van Voorhies WA, Ward S. Broad oxygen tolerance in the nematode Caenorhabditis elegans. J Exp Biol. 2000;203(Pt 16):2467–78. Epub 2000/07/21. Division of Parasitology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, Miyazaki, Japan The Aquatic Gardener 10 (1997): 126-131 Water Plants 101: Dave Huebert A. M. Humphrey, “Chlorophyll as a color and functional ingredient,” Journal of Food Science, vol. 69, no. 5, pp. 422–425, 2004. Chlorophyll Extraction from Microalgae: A Review on the Process Engineering Aspects Aris Hosikian, Su Lim, Ronald Halim, and Michael K. Danquah Bio Engineering Laboratory (BEL), Department of Chemical Engineering, Monash University, Victoria (2010) “Fluorescence measurements of free Mg2” Elisabeth M Froschauer, Martin Kolisek, Frank Dieterich, Monika Schweigel, Rudolf J Schweyen, Max F Perutz Laboratories Department of Microbiology and Genetics University of Vienna Campus Vienna Biocenter, Dr Bohrgasse 9 A1030 Vienna Austria “Water content quantitatively affects metabolic rates over the course of plant ontogeny” State Key Laboratory of Grassland AgroEcosystem School of Life Sciences Lanzhou University Lanzhou 730000 China, College of Forestry Southwest Forestry University Bailongsi China, Department of Biological Sciences Missouri University of Science and Technology, School of Integrative Plant Science Plant Biology Section Cornell University “Determination of elements in algae by different atomic spectroscopic methods” (MG2++ and Carbonic Acid analysis) Attila József University Department of Inorganic and Analytical Chemistry Szeged Hungary, Albert Szent Györgyi Medical University Department of Pharmacognosy Szeged Hungary, School of Pharmacy Portsmouth Polytechnic Portsmouth UK Studies on Sonication of Plants for Accelerated Growth: Sonication and ultrasound: Impact on plant growth and development Beneficial effects of ultrasound on plants—a review - ScienceDirect TOVATECH Tovatech - The Source for Ultrasonic Cleaners & Lab Equipment for More than 10 Years. Greenhouse Product News Effects of Blue Light on Plants Erik Runkle FEBRUARY 2017 Ultrasound has been investigated as a control for zoo[1] plankton in ballast water. Holm et al. (2008) tested ultrasound effects on a cladoceran (Ceriodaphnia dubia), rotifers (Brachionus plicatilis, B. calyciflorus, and Philodina sp.), and brine shrimp (Artemia sp.). The Potential Use of Ultrasound to Control the Trematode Bolbophorus confusus by Eliminating the Ram's Horn Snail Planorbella trivolvis in Commercial Aquaculture Settings Bradley T. Goodwiller & James P. Chambers, Pages 485-488 Sep 2012 Ultrasound has been investigated as a method for zebra mussel (Dreissena polymorpha) control. (Kowalewski et al. 1993, Donskoy and Ludiyanskiy 1995). Donskoy and Ludiyanskiy (1995) cite research in which ultrasound ranging from 20kHz to 380kHz was used to induce cavitation and mortality in veliger, juvenile, and adult zebra mussels. The Potential Use of Ultrasound to Control the Trematode Bolbophorus confusus by Eliminating the Ram's Horn Snail Planorbella trivolvis in Commercial Aquaculture Settings Bradley T. Goodwiller & James P. Chambers Pages 485-488 | Received 27 Jan 2012, Accepted 09 Apr 2012, Published online: 10 Sep 2012 Ultrasonics Sonochemistry Volume 13, Issue 5, July 2006, Pages 446-450 Ultrasonics Sonochemistry Ultrasonic frequency effects on the removal of Microcystis aeruginosa 2005.09.012 Effects of ultrasound irradiation on the growth of Japanese radish sprouts. Marie Tabaru[1], Ryusuke Fujino and Kentaro Nakamuray Electrical Stimulation and Effects on Plant Growth in Hydroponics Bonghwan Kim and Kyunghan Chun University of Daegu Solar-Powered Electroculture Technique For Backyard Farming Elenor M. Reyes, GlennJordan M. Achico Accelerated Plant Growth--https://www.motherearthnews.com/organic-gardening/plant-growth-zmaz84mjzloeck/ Molecular strategies for improving waterlogging tolerance in plants E.S. Dennis, R. Dolferus, M. Ellis, M. Rahman, Y. Wu, F.U. Hoeren, A. Grover, K.P. Ismond, A.G. Good, W.J. Peacock Journal of Experimental Botany, Volume 51, Issue 342, January 2000, Pages 89–97 Determination of elements in algae by different atomic spectroscopic methods. Chlorophyll Destruction by the Bisulfite-Oxygen System' 1977-PEISER/YANG Department of Vegetable Crops, University of California, Davis, California Professor Johnjoe McFadden's Presentation "Does Biology Need Quantum Mechanics?" at Imperial College London's Festival of Science 2014

Hi all - 2 weeks into my plant-only cycle, and am getting a nice primordial ooze going with brown algae/diatoms and driftwood biofilm. I'm not sure if that's some sort of hair algae or just a mix if diatoms and biofilm. Tanks are 10 and 20 gallons. No ammonia, around 20 nitrates and around 1ppm nitrite, 7.5-7.7ph. Fairly hard water, medium buffer. I'm wondering if it's time to chuck some snails in. Would love an Amono but am guessing they wouldn't do so well yet. Any tips/advise? Not sure if I should leave it be or not.

I am seeing slight improvement. I did 50% water change, rinsed filter media, and lowered lights. 1st pic is right after, others are 4 days later. My tank is a 60 tall so I am concerned about not enough light for the bottom, I was thinking of higher light and siesta time. I do have a bristlenose in quarantine for this tank.