Patent Publication Number: US-11382332-B2

Title: Methods and systems for controlling invasive mussel species

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     The present application is a continuation-in-part application of U.S. patent application Ser. No. 16/271,523, filed Feb. 8, 2019, entitled METHODS AND SYSTEMS FOR CONTROLLING INVASIVE MUSSEL SPECIES, which is itself a Continuation-in-part application of U.S. patent application Ser. No. 16/117,819, filed Aug. 30, 2018, entitled METHODS AND SYSTEMS FOR CONTROLLING INVASIVE MUSSEL SPECIES, each of which is incorporated by reference in their entireties herein. 
    
    
     BACKGROUND OF THE INVENTION 
     Field of the Invention 
     The present invention is generally directed to chlorophyll water treatments effective for killing or preventing the spread of invasive mussel species. 
     Description of the Prior Art 
       Dreissenid  mussels, also known as Zebra Mussels &amp; Quagga Mussels, are one of the most biological invasive species in North American waters. Zebra mussels are notorious for their biofouling capabilities by colonizing water supply pipes of hydroelectric and nuclear power plants, public water supply plants, and industrial facilities. They colonize pipes constricting flow, therefore reducing the intake in heat exchangers, condensers, fire-fighting equipment, and air conditioning and cooling systems. Navigational and recreational boating can be affected by increased drag due to attached mussels. Small mussels can get into engine cooling systems causing overheating and damage. Navigational buoys have been sunk under the weight of attached zebra mussels. Fishing gear can be fouled if left in the water for long periods. Deterioration of dock pilings has increased when they are encrusted with zebra mussels. Continued attachment of zebra mussels can cause corrosion of steel and concrete affecting its structural integrity. 
     Containment of invasive mussels can be difficult and cost prohibitive, particularly when the volume of water to be treated is large. For example, a paper company plant located on Lake Michigan spent $1.4 million for removal of zebra mussels from 400 cubic yards near plant equipment. Some current methods of controlling invasive mussels include chemical, bromine, quaternary and poluquaternary ammonium compounds, aromatic hydrocarbons, copper (heavy metal), endothall, zequanox, coatings, UV light, filtration, mechanical, and operational. 
     What is needed is an improved, low-cost method of treating and controlling the spread of invasive mussel species. 
     SUMMARY OF THE INVENTION 
     In one embodiment of the present invention, there is provided a method of controlling the spread of an invasive mussel species, and particularly a  Dreissenid  mussel species. The method comprises introducing a nonnative source of chlorophyll to a body of water comprising the invasive mussel species. The nonnative source of chlorophyll is introduced at an amount sufficient to provide a concentration of chlorophyll in the body of water of at least about 10 μg/L. The body of water has a pH of about 7 to about 8 after introducing the nonnative source of chlorophyll. 
     In another embodiment, there is provided a method of controlling the spread of an invasive  Dreissenid  mussel species. The method comprises introducing a nonnative source of chlorophyll to a body of water comprising the invasive mussel species. The nonnative source of chlorophyll is introduced at an amount sufficient to provide a concentration of chlorophyll in the body of water of at least about 10 μg/L. The nonnative source of chlorophyll comprises a material selected from the group consisting of cyanobacteria, alfalfa, soy beans, wheat grass, wheat straw, barley grass, mulberry, chlorella, sea weed, fresh water moss, spirulina, and mixtures thereof. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a graph showing measured chlorophyll concentrations at various amounts of alfalfa powder added to distilled water; 
         FIG. 2  is a graph showing measured chlorophyll concentrations at various amounts of spirulina powder added to distilled water; 
         FIG. 3  is a graph showing the amount of alfalfa powder needed to obtain chlorophyll concentrations by extrapolation beyond the measurable limit using the trendline formed using the data points shown in  FIG. 1 ; and 
         FIG. 4  is a graph showing the amount of spirulina powder needed to obtain chlorophyll concentrations by extrapolation beyond the measurable limit using the trendline formed using the data points shown in  FIG. 2 . 
     
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT 
     The present invention is generally directed to methods and systems for killing, preventing, or inhibiting the growth and spread of invasive  Dreissenid  mussel species, such as Zebra mussels ( Dreissena polymorpha ) and Quagga mussels ( Dreissena rostriformis bugensis ), by treating bodies of water comprising the mussels with a nonnative source of chlorophyll. Chlorophyll is a group of green pigments present in cyanobacteria and in the chloroplasts of plants and algae that absorb light for photosynthesis. There are five types of chlorophyll: chlorophyll a, which is present in all photosynthetic organisms except bacteria; chlorophyll b, in plants and green algae; and chlorophylls c, d and e, in some algae. As used herein, the term “nonnative” refers to a source of chlorophyll that is not indigenous or naturally present in the body of water being treated. This can include aquatic and nonaquatic plants and/or other organisms (indigenous or otherwise) that have been artificially modified or processed (e.g., pulverized, blended, liquified, etc.) so as to be in a non-natural form before being introduced to the body of water. For example, leaves, stems, and/or other plant parts may be harvested from aquatic and/or nonaquatic plants and artificially processed to form the nonnative source of chlorophyll. In certain embodiments, the nonnative source of chlorophyll comprises a material selected from the group consisting of cyanobacteria, alfalfa, soy beans, wheat grass, wheat straw, barley grass, mulberry, chlorella, sea weed, fresh water moss, spirulina, animalia feces, and mixtures thereof. In particularly preferred embodiments, the nonnative source of chlorophyll comprises a material collected from the body of water to be treated. In certain embodiments, the nonnative source of chlorophyll is provided in a form selected from the group consisting of vapor, liquid, paste, powder, pellets, cubes, blocks (both immediate or time release), animalia food, and combinations thereof. In particularly preferred embodiments, the source of chlorophyll is in liquid form. In particularly preferred embodiments, the source of chlorophyll is in powder form (either dry powder or suspended in liquid). 
     In certain preferred embodiments, the nonnative source of chlorophyll comprises an aquatic or nonaquatic plant (including algae species) or plant part that has been artificially processed. In such embodiments, certain considerations should be made during processing of the plant or plant part so as to release the aroma and taste of the plant while retaining its fibers and/or oils. In certain embodiments, the source of chlorophyll comprises processed plant particles (e.g., powder particles) having an average diameter of about 0.05 μm to about 650 μm, preferably about 0.2 μm to about 500 μm, and more preferably about 40 μm to about 400 μm. Without being bound by any theory, it is believed particles sizes within these ranges allow for normal ingestion and filtration by the mussels. The size can be determined at the time of application or after dissolving or breaking up when in contact with a liquid or water environment. 
     Flavoring and other additives may also be mixed with the nonnative source of chlorophyll, for example to encourage drinking by birds and other animalias or to impart preferred consistency and texture. In certain embodiments, the method further comprising introducing a flavoring along with the nonnative source of chlorophyll. The flavoring may be natural or synthetic. The flavoring can be introduced together with or separately from the nonnative source of chlorophyll and may similarly be provided in the form of vapor, liquid, paste, powder, pellets, cubes, blocks, and combinations thereof. In certain preferred embodiments, the flavoring comprises a natural flavoring, such as processed legume or foliage aroma (e.g., from soybean or clover), blended fish, and/or fish meal, that has been mixed with the nonnative source of chlorophyll to be introduced to the body of water. In certain embodiments, soured or spoiled flavoring sources should be avoided. Particularly preferred sources of flavoring include chlorophyll-a, which is believed to be particularly desirable to mussels. When flavoring is used, the invasive mussels can be killed at a faster rate. Without being bound by any theory, it is believed that the flavoring increases activity and intake of nutrients, including chlorophyll, by the mussels, which leads to increased blockage and suffocation by the filter feeding invasive mussels. Additionally, the nonnative source of chlorophyll may be mixed with natural or synthetic binders, such as cellulose, lignin, and/or other polymer binders. 
     In certain embodiments, a fibrous material is mixed with the nonnative source of chlorophyll. In certain preferred embodiments, the fibrous material is a cellulosic fibrous material. The fibrous material may be derived from a processed aquatic or nonaquatic plant, which can be the same plant providing the source of chlorophyll or a different plant. Other natural or synthetic fibrous materials may also be used. In certain embodiments, the fibrous material comprises fibers having an average length of about 0.05 μm to about 650 μm, preferably about 0.2 μm to about 500 μm, and more preferably about 40 μm to about 400 μm. 
     Methods of controlling the invasive mussel species comprise introducing the nonnative source of chlorophyll to a body of water comprising the invasive mussel species. The body of water may be an open or closed water system and may be internal or external environment. The body of water may include, but is not limited to, lakes, reservoirs, ponds, streams, rivers, processing facilities, and aquariums. As used herein, the “body of water” may refer to the entirety of the water contained within the system or only a localized portion of the water within the system, for example a localized portion of the water in the surrounding water proximate to the invasive mussel species. The body of water may also comprise one or more structures submerged in the body of water including, but not limited to, piping, pumps, inlets, outlets, ballast, piers, boats, and other structures both manmade and nature made. The body of water may also comprise natural and non-natural structures including, but not limited to, wood, cement, hard surfaces, mud, and remains of other mussels. Invasive mussel species often aggregate and grow on one or more of the above-noted structures in the body of water, and thus the nonnative source of chlorophyll is preferably introduced to the body of water at a location proximate to such structures. 
     The nonnative source of chlorophyll may be introduced to the body of water by a variety of methods including, for example, pouring, spreading, or spraying the nonnative source of chlorophyll onto the surface of the body of water, or injecting the nonnative source of chlorophyll into the body of water below the surface. In other embodiments, for example when the nonnative source of chlorophyll is provided as an animal food, the nonnative source of chlorophyll may be introduced to the body of water by feeding the nonnative source of chlorophyll to an animalia and allowing the animalia to defecate at a location proximate to the invasive mussel species. 
     The amount of nonnative chlorophyll introduced to the body of water can depend on a number of factors. In certain embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide a chlorophyll concentration in the body of water at a concentration of at least about 10 μg/L. In certain embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide a chlorophyll concentration in the body of water at a concentration of about 10 μg/L to about 900 μg/L, preferably about 15 μg/L to about 50 μg/L. However, in certain embodiments, much higher chlorophyll concentrations may be used. In certain such embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide a chlorophyll concentration in the body of water at a concentration up to about 10,000 μg/L, up to about 16,000 μg/L (particularly when biomass cleanup is not a concern), up to about 50,000 μg/L, up to about 100,000 μg/L, up to about 130,000 μg/L (particularly when faster eradication is desired and/or little to no consideration for aquatic life and/or little to no consideration for residue in equipment), up to about 160,000 μg/L, or up to about 200,000 μg/L. In certain embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide a chlorophyll concentration in the body of water at a concentration of about 10 μg/L to about 200,000 μg/L, preferably about 15 μg/L to about 160,000 μg/L. It should be understood that even higher chlorophyll concentrations may be used, depending on the particular application and solubility of the source of chlorophyll. 
     Additionally, the chlorophyll concentration and duration of the treatments can be selected so as to provide effective short-term or long-term kill. In certain preferred embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide an average chlorophyll concentration in the body of water over the duration of treatment of about 15 μg/L to about 25 μg/L for about 3 to about 4 days. In certain preferred embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide an average chlorophyll concentration in the body of water over the duration of treatment of about 20 μg/L to about 40 μg/L for about 1 to about 2 days. In certain preferred embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide an average chlorophyll concentration in the body of water over the duration of treatment of at least about 40 μg/L for less than about 24 hours. Saturated concentrations of chlorophyll may also be used, depending on factors such as the presence of aquatic life and urgency of remedy. For example, the nonnative source of chlorophyll may be introduced so as to provide upper hypereutrophic levels of chlorophyll when aquatic life is not of concern. Additionally, in highly infested bodies of water (i.e., with a large number of zebra mussels within an area), greater concentrations of chlorophyll may be needed to achieve sufficient kill. Therefore, in certain embodiments, the nonnative chlorophyll is introduced to the body of water so as to provide a chlorophyll concentration in the body of water at a concentration of at least about 100 μg/L, at least about 200 μg/L, at least about 500 μg/L, at least about 1,000 μg/L, at least about 10,000 μg/L, at least about 16,000 μg/L, at least about 50,000 μg/L, at least about 100,000 μg/L, at least about 130,000 μg/L, at least about 160,000 μg/L, or at least about 200,000 μg/L. 
     The efficacy of the treatment methods can be dependent on several variances including, but not limited to, water temperature, salinity, toxins, oxygen level, age of mussels, current flow, turbidity, and pH. For example, older mussels can be killed at lower chlorophyll doses. The turbidity and pH of the water can impact both the efficacy of controlling invasive mussels as well as the survival of desirable aquatic life. In certain embodiments, the turbidity of the body of water is maintained at about 5 to about 500 ppm. In certain embodiments, and pH of the water is maintained at about 7 to about 8. 
     As noted above, the nonnative source of chlorophyll can comprise a material collected from the body of water to be treated. Therefore, in certain embodiments, the methods further comprise collecting an aquatic plant and/or algae material from the body of water and artificially processing the material before introducing the material into the body of water as the nonnative source of chlorophyll. For example, in certain such embodiments, the collected aquatic plant and/or algae is ground or blended into particulates. This artificial processing advantageously allows for increased exposure of the chlorophyll content within the plant or algae when re-introduced into the body of water. The artificial processing may further comprising mixing the material with flavorings or other additives, as discussed above, before re-introducing the material to the body of water. 
     In certain embodiments, the method further comprises monitoring the chlorophyll concentration of the body of water. The monitoring can be performed with a permanent monitor (e.g., installed at a permanent location in the body of water) or by intermittent manual readings. Monitoring the chlorophyll concentration at the body of water can be used to determine the amount and intervals of treatments with the nonnative source of chlorophyll to maintain a predetermined chlorophyll concentration to effect kill or prevention of the invasive mussel species. 
     Embodiments of the present invention are also directed to systems for controlling the growth and spread of invasive mussel species. The systems generally comprise a dosing station configured to introduce the nonnative source of chlorophyll (and any additional components described above) to the body of water. In certain embodiments, the system further comprises a chlorophyll concentration monitor residing in the body of water and configured to measure the concentration of chlorophyll at a location in the body of water. The dosing station can comprise a reservoir for storing the nonnative source of chlorophyll and an outlet for introducing the nonnative source of chlorophyll to the body of water. The outlet can be configured to introduce the nonnative source of chlorophyll, for example, by pouring, spreading, or spraying the nonnative source of chlorophyll onto the surface of the body of water, or injecting the nonnative source of chlorophyll into the body of water below the surface. The outlet may also be configured to provide an animalia feed comprising the nonnative source of chlorophyll to or around the body of water for consumption by animalias such as fish and waterfowl. The monitor may be any of a variety of chlorophyll concentration monitors known in the art. The system may further comprise a controller in communication with both the monitor and dosing station. In use, the controller instructs the dosing station to introduce an amount of the nonnative source of chlorophyll to the body of water so as to maintain the chlorophyll concentration in the body of water at a predetermined level as measured by the monitor. The dosing stations can be located anywhere that mussel kill or prevention is desired. In particular embodiments, the dosing station is located at one or more fresh water inlets. This allows chlorophyll concentrations to maintain necessary high levels where lower levels of chlorophyll are generally present. 
     The particular form of the nonnative source of chlorophyll can be selected based on certain advantages for certain applications. For example, artificially modified or processed aquatic plants (e.g., grass or leaf plants) and/or algae may be advantageous so as to not inadvertently introduce other potentially invasive organisms to the body of water. Additionally, a plant leaf source compressed into a formation releases at a slower rate than powder or liquid forms, and thus such a source may be advantageous for slow-release applications. Spirulina and chlorella have a stronger affinity for osmosis into water than grasses or leaf substances with, and leaf substances seem to be eaten by fish more readily. Thus, in particular embodiments, the nonnative source of chlorophyll comprises spirulina and/or chlorella. In certain embodiments, the nonnative source of chlorophyll excludes grass and/or leaf plants. Chlorella has a higher concentration of chlorophyll than spirulina, which can be advantageous for closed areas, such as piping and equipment. Additionally, spirulina is larger in size than chlorella. Therefore, in certain embodiments, the nonnative source of chlorophyll comprises a mixture of chlorella (for smaller mussels) and spirulina (for larger particles and larger mussels). This mixture may also comprise a leaf source (such as alfalfa) that aquatic life readily consumes. Such a mixture can provide a multi-faceted approach to controlling the spread of invasive mussels. Mixtures of spirulina and alfalfa pellets were particularly advantageous for treatments in areas with aquatic life, as fish, snails, and insects can survive in high chlorophyll levels (30 μg/L-35 μg/L) when such mixtures are used. 
     In certain embodiments, the nonnative source of chlorophyll does not comprise heavy metals, such as copper, commonly used in prior art treatment methods. Heavy metals can be precursors to several neurological conditions and concentrations can increase over time in bodies of water treated with prior art methods. Therefore, in certain embodiments, the methods and systems advantageously do not comprise introducing a heavy metal to the body of water. Additionally, the use of pesticides has known and possibly unknown health risks to humans. Therefore, in certain embodiments, the methods and systems advantageously do not comprise introducing a pesticide to the body of water. 
     Additional advantages of the various embodiments of the invention will be apparent to those skilled in the art upon review of the disclosure herein and the working examples below. It will be appreciated that the various embodiments described herein are not necessarily mutually exclusive unless otherwise indicated herein. For example, a feature described or depicted in one embodiment may also be included in other embodiments, but is not necessarily included. Thus, the present invention encompasses a variety of combinations and/or integrations of the specific embodiments described herein. 
     As used herein, the phrase “and/or,” when used in a list of two or more items, means that any one of the listed items can be employed by itself or any combination of two or more of the listed items can be employed. For example, if a composition is described as containing or excluding components A, B, and/or C, the composition can contain or exclude A alone; B alone; C alone; A and B in combination; A and C in combination; B and C in combination; or A, B, and C in combination. 
     The present description also uses numerical ranges to quantify certain parameters relating to various embodiments of the invention. It should be understood that when numerical ranges are provided, such ranges are to be construed as providing literal support for claim limitations that only recite the lower value of the range as well as claim limitations that only recite the upper value of the range. For example, a disclosed numerical range of about 10 to about 100 provides literal support for a claim reciting “greater than or equal to about 10” (with no upper bounds) and a claim reciting “less than or equal to about 100” (with no lower bounds). 
     EXAMPLES 
     The following examples set forth various specific treatment methods for killing  Dreissenid  mussel colonies collected from an infested lake. It is to be understood, however, that these examples are provided by way of illustration and nothing therein should be taken as a limitation upon the overall scope of the invention. 
     Items and brands used for testing that were bought were: Chlorella, NOW Foods, Alfalfa Powder, NOW Foods, Certified Organic Wheat Grass, NOW Foods, Organic Spirulina Powder, Sunny Green Products, Barley Grass Bulk Powder, Nature&#39;s Way Brands, LLC, Wheat Grass Tablets, Sunny Green Products, Fresh Alfalfa Hay, Fresh Soy Bean Plants and Wheat Straw. 
     During the tests below, readings of chlorophyll concentration were taken 30 to 60 minutes after adding substances to either lake water or distilled water, unless indicated otherwise. 
     Example I 
     Locating a small body of water with an infestation of zebra Mussels, a rock was collected having a colony thereon and placed in an aquarium with fish. After a couple days, the water had become extremely clear, which was different than when 10 gallons of water was originally retrieved from the body of water where the zebra mussels were retrieved. The mussels were fed a chlorophyll-based food, spirulina, by adding a heaping table spoon into the 10-gallon fish tank. The amount of spirulina was enough to turn the water dark/bright green. A few days later, the mussels were open (indicating death). At 7 days, the tank was a smell of worsening debris. The fish had died, likely due to the sudden change in the living environment. The water was drained, and the sample drenched in high concentration of bleach until the shells were bleached out. 
     Example II 
     Additional colonies of zebra mussels were collected and fed several other plant-based products comprising chlorophyll. Chlorophyll sources tested included: wheat grass, alfalfa powder, barley grass, chlorella, dried soy bean leaves, dried alfalfa leaves, and Spirulina. All zebra mussels died similar to Example I above. It was determined that the chlorophyll of the plant-based products was killing the zebra mussels. 
     Example III 
     Chlorophyll levels were measured at locations in bodies of water that were known to be completely infested, partially infested, and not infested (thought to be non-infested because no boating activity allowed). Fluorescence readings taken to estimate chlorophyll levels were as follows:
         i) complete infestation at 3 μg/L;   ii) broadly infested but incomplete at 7 μg/L;   iii) one lake infestation near inlet but not the shoreline of the bowl of the lake at 15 μg/L to 25 μg/L (with smell); and   iv) no apparent infestation at 17 μg/L.
 
Based on this, it was concluded that chlorophyll levels can be used to determine likelihood of mussels spreading or presence. Notably, gold fish living with dead zebra mussels more than doubled in size within 6 weeks. Therefore, it is believed that aquatic life will flourish where mussel control is initiated.
       

     Example IV 
     Further lab testing was performed to measure the effect of chlorophyll concentration on killing zebra mussel colonies. A Fluorosense Meter from Turner Designs was used to determine the actual readings that had been used previously by measuring both grain weight and μg/L presence of chlorophyll. Material used to calibrate was also obtained, Fluorosense Chlorophyll Standard Solution, provided by Turner Designs, P/N 2860-220. Equipment included: 1-50-gallon aquarium, 1-10-gallon aquarium, 40×-600× microscope, 3.5×-90× microscope, hydrometer, test tubes, numerous air and water pumps, precision (grain) and bulk pounds&#39; scales, food blenders, ECO Testr Turbidity Meter, HM Digital pH Meter, Turner Designs Flurosense—Chlorophyll Meter, Microscope Camera, 22 1-gallon fish bowls, 15 5-gallon buckets, 2 65-gallon barrels and numerous miscellaneous equipment. 
     Several tests were performed by measuring weights of products to determine death prior to receiving equipment to evaluate the estimated chlorophyll levels in μg/L, giving the bases to follow up on using the meter readings. The chlorophyll concentrations and turbidity resulting from 12 grain (weight) of various products added to 2 liters of distilled water are provided in Table 1 below. 
     
       
         
           
               
               
               
               
             
               
                   
                 TABLE 1 
               
             
            
               
                   
                   
               
               
                   
                   
                   
                 Morning 
               
               
                   
                   
                   
                 (Approximately 12 
               
               
                   
                 Start 
                 1 Hour 
                 hours later) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Chlorophyll 
                 Turbidity 
                 Chlorophyll 
                 Turbidity 
                 Chlorophyll 
                 Turbidity 
               
               
                 Product 
                 μg/L 
                 PPM 
                 μg/L 
                 PPM 
                 μg/L 
                 PPM 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Spirulina 
                 113 
                 10 
                 102 
                 10 
                 84 
                 10 
               
               
                 Alfalfa 
                 66 
                 20 
                 22 
                 20 
                 13 
                 20 
               
               
                 Powder 
               
               
                 Wheat 
                 45 
                 20 
                 25 
                 10 
                 15 
                 10 
               
               
                 Grass 
               
               
                 Powder 
               
               
                 Barley 
                 35 
                 20 
                 25 
                 20 
                 15 
                 20 
               
               
                 Grass 
               
               
                 Powder 
               
               
                 Chlorella 
                 25 
                 10 
                 26 
                 10 
                 22 
                 10 
               
               
                   
               
            
           
         
       
     
     Various sources of chlorophyll were tested at various concentration ranges by placing live colonies of zebra mussels in 1-gallon containers filled with either lake water (having average chlorophyll concentration of 1 to 3 μg/L) or distilled water. The source of chlorophyll was then added to the container and dissolved. The time required to kill the zebra mussel colonies was determined by tapping the open mussels and observing the response. The results are provided in Tables 2-9, below. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Infested Lake Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                 Alfalfa Powder 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                 Alfalfa Fresh 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                 Blended 
               
               
                 Soy Bean Fresh 
                 12 μg/L 
                 22 μg/L 
                 &lt;96 
               
               
                 Blended 
               
               
                 Barley Grass 
                 12 μg/L 
                 22 μg/L 
                 &lt;96 
               
               
                 Powder 
               
               
                 Wheat Grass 
                 12 μg/L 
                 22 μg/L 
                 &lt;96 
               
               
                 Powder 
               
               
                 Chlorella 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Distilled Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                 Alfalfa Powder 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                 Alfalfa Fresh 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                 Blended 
               
               
                 Soy Bean Fresh 
                 12 μg/L 
                 22 μg/L 
                 &lt;96 
               
               
                 Blended 
               
               
                 Barley Grass 
                 12 μg/L 
                 22 μg/L 
                 &lt;96 
               
               
                 Powder 
               
               
                 Wheat Grass 
                 12 μg/L 
                 22 μg/L 
                 &lt;96 
               
               
                 Powder 
               
               
                 Chlorella 
                 12 μg/L 
                 22 μg/L 
                 &lt;72 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Infested Lake Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 22 μg/L 
                 35 μg/L 
                 &lt;48 
               
               
                 Alfalfa Powder 
                 22 μg/L 
                 35 μg/L 
                 &lt;48 
               
               
                 Alfalfa Fresh 
                 22 μg/L 
                 35 μg/L 
                 &lt;48 
               
               
                 Blended 
               
               
                 Soy Bean Fresh 
                 22 μg/L 
                 35 μg/L 
                 &lt;72 
               
               
                 Blended 
               
               
                 Barley Grass 
                 22 μg/L 
                 35 μg/L 
                 &lt;72 
               
               
                 Powder 
               
               
                 Wheat Grass 
                 22 μg/L 
                 35 μg/L 
                 &lt;72 
               
               
                 Powder 
               
               
                 Chlorella 
                 22 μg/L 
                 35 μg/L 
                 &lt;48 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Distilled Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 22 μg/L 
                 35 μg/L 
                 &lt;48 
               
               
                 Alfalfa Powder 
                 22 μg/L 
                 35 μg/L 
                 &lt;48 
               
               
                 Alfalfa Fresh 
                 22 μg/L 
                 35 μg/L 
                 &lt;48 
               
               
                 Blended 
               
               
                 Soy Bean Fresh 
                 22 μg /L 
                 35 μg /L 
                 &lt;72 
               
               
                 Blended 
               
               
                 Barley Grass 
                 22 μg /L 
                 35 μg /L 
                 &lt;72 
               
               
                 Powder 
               
               
                 Wheat Grass 
                 22 μg /L 
                 35 μg /L 
                 &lt;72 
               
               
                 Powder 
               
               
                 Chlorella 
                 22 μg /L 
                 35 μg /L 
                 &lt;48 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Infested Lake Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 35 μg/L 
                 60 μg/L 
                 &lt;24 
               
               
                 Alfalfa Powder 
                 35 μg/L 
                 60 μg/L 
                 &lt;24 
               
               
                 Chlorella 
                 35 μg/L 
                 60 μg/L 
                 &lt;24 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Distilled Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 35 μg/L 
                 60 μg/L 
                 &lt;24 
               
               
                 Alfalfa Powder 
                 35 μg/L 
                 60 μg/L 
                 &lt;24 
               
               
                 Chlorella 
                 35 μg/L 
                 60 μg/L 
                 &lt;24 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Infested Lake Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 60 μg/L 
                 199+ μg/L 
                 ≈8 
               
               
                 Alfalfa Powder 
                 60 μg/L 
                 199+ μg/L 
                 ≈8 
               
               
                 Chlorella 
                 60 μg/L 
                 199+ μg/L 
                 ≈8 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Distilled Water. 
               
            
           
           
               
               
               
            
               
                 Source of 
                   
                 Time to Death 
               
               
                 Chlorophyll 
                 Chlorophyll Concentration Range 
                 (hours) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Spirulina 
                 60 μg/L 
                 199+ μg/L 
                 ≈8 
               
               
                 Alfalfa Powder 
                 60 μg/L 
                 199+ μg/L 
                 ≈8 
               
               
                 Chlorella 
                 60 μg/L 
                 199+ μg/L 
                 ≈8 
               
               
                   
               
            
           
         
       
     
     It should be considered that fish are able to tolerate higher levels of chlorophyll if introduced through a time period, instead of a sudden influx to reach the chlorophyll level for zebra mussel death. Additionally, it was observed in the study above that leaf plants seem to have a greater impact on controlling mussels than grasses. Leaf plants are considered a benefit to aquatic life because leave plants are most often considered better food sources for fish. 
     Example V 
     Some prior art has suggested that zebra mussels may be killed by ducks and geese, for example by ingesting the mussels. Visual observation was performed along lake shores where water fowl moved their habitat around the lake because of grass and clover. The zebra mussels were dead in the areas of travel, but not in the immediate vicinity of the waterfowl. Notably, the shells of the zebra mussels were still intact and attached. This would suggest that the mussels were not eaten by the waterfowl. Additionally, the zebra mussel deaths did not appear due to foot traffic of the waterfowl because the depth of water exceeded the capability of foot contact. Further observation noticed a green tint to the defecation left on the shore. Based on these observations, waterfowl defecation was tested for chlorophyll levels and use for killing zebra mussel colonies. 
     Fresh waterfowl defecation was collected and placed in distilled water to discover readings of chlorophyll above 199 μg/L. Placing zebra mussels in this water resulted in death within less than 24 hours. See Table 10 below. 
     
       
         
           
               
               
               
               
               
               
             
               
                 TABLE 10 
               
               
                   
               
               
                   
                   
                   
                   
                   
                 Time To 
               
               
                   
                   
                   
                   
                   
                 Death As 
               
               
                   
                   
                 Concen- 
                   
                   
                 Determined 
               
               
                   
                   
                 tration 
                   
                   
                 by Tapping 
               
               
                   
                   
                 in Distilled 
                 Tur- 
                   
                 Open 
               
               
                 Source 
                 Notes 
                 Water 
                 bidity 
                 pH 
                 Mussels 
               
               
                   
               
             
            
               
                 Duck 
                 Approximately 
                 &gt;199 μg/L 
                 350 ppm 
                 7.5 
                 &lt;24 Hours 
               
               
                 Defecation 
                 ⅛ cup in 
               
               
                 Dry 
                 gallon of water 
               
               
                   
               
            
           
         
       
     
     Following the results above, additional waterfowl defecation was collected. The dried defecation was dissolved until a 40 μg/L estimated chlorophyll concentration was obtained. At that time, a rock with a colony of zebra mussels was placed in the body of water, along with a bubble stone for an oxygen source. Within 20 hours, the zebra mussel population was dead. See Table 11. 
     
       
         
           
               
               
               
               
               
               
             
               
                 TABLE 11 
               
               
                   
               
               
                   
                   
                   
                   
                   
                 Time To 
               
               
                   
                   
                   
                   
                   
                 Death As 
               
               
                   
                   
                 Concen- 
                   
                   
                 Determined 
               
               
                   
                   
                 tration 
                   
                   
                 by Tapping 
               
               
                   
                   
                 in Distilled 
                 Tur- 
                   
                 Open 
               
               
                 Source 
                 Notes 
                 Water 
                 bidity 
                 pH 
                 Mussels 
               
               
                   
               
             
            
               
                 Duck 
                 One dropping 
                 40 μg/L 
                 270 ppm 
                 7.8 
                 &lt;24 Hours 
               
               
                 Defecation 
                 approximately 
               
               
                 Dry 
                 2″ long, in 
               
               
                   
                 gallon of 
               
               
                   
                 water 
               
               
                   
               
            
           
         
       
     
     Notably, spiral snails collected while collecting zebra mussels had no response to the higher chlorophyll levels. Passing CO 2  gases through an air stone into the chlorophyll concentrations did not have any effect on the chlorophyll readings. 
     Example VI 
     A quart container of a mixture of alfalfa pellets, alfalfa powder, spirulina powder and sugar was placed in a lake near shore line. Weather conditions were windy with water craft waves hitting the shore line. Later in the day (approximately 2 hours), rain fell. The test was still considered successful, as the zebra mussels in the immediate area were found to be unresponsive 7 hours later and still unresponsive 30 hours later. This suggests that high saturation for short periods of time can be effective for killing zebra mussels. 
     Example VII 
     Additional lab testing was performed with water extracted from a lake. Water extracted from the lake with chlorophyll concentrations averages of 20 μg/L to 25 μg/L resulted in death in less than 48 hours, although the chlorophyll concentration level had to be rejuvenated as the chlorophyll levels dropped during the period. Readings taken over the 48-hour period were: 
     25 μg/L (lake water); 
     9 μg/L (24 hours); 
     20 μg/L (reading after low levels of chlorophyll was replaced); and 
     7 μg/L (at the end of 48 hours). 
     This resulted in death in less than 48 hours after introduction. Based on this, it is believed the chlorophyll concentration can determine an estimated time for death, for example: 15-25 μg/L for 3 to 4 days of exposure; 20-40 μg/L 1 to 2 days of exposure; and above 40 μg/L for less than 24 hours. 
     Example VIII 
     Initial observations in Example VII were run using lake water, but there was some concern with variations in results. Tests were run with both lake water and distilled water to confirm consistency and also to confirm that the combination of material in the lake water was not creating an unknown chemical reaction that could be killing the ZMs. Therefore, a similar study was performed in distilled water and a comparison found that distilled water provided the same results. 
     Example IX 
     Blended fresh soy bean leaves were added to distilled water at an estimated concentration of 150 μg/L &amp; 10 ppm turbidity. A colony of zebra mussels were added attached to a rock. After 24 hours, there was response from partially opened zebra mussels. Time to kill was approximately 36 hours. See Table 12. 
     
       
         
           
               
               
               
               
               
               
             
               
                 TABLE 12 
               
               
                   
               
               
                   
                   
                   
                   
                   
                 Time To 
               
               
                   
                   
                   
                   
                   
                 Death As 
               
               
                   
                   
                 Concen- 
                   
                   
                 Determined 
               
               
                   
                   
                 tration 
                   
                   
                 by Tapping 
               
               
                   
                   
                 in Distilled 
                 Tur- 
                   
                 Open 
               
               
                 Source 
                 Notes 
                 Water 
                 bidity 
                 pH 
                 Mussels 
               
               
                   
               
             
            
               
                 Fresh 
                 Blended in 
                 150 μg/L 
                 10 ppm 
                 7.4 
                 ≈36 hours 
               
               
                 Ground 
                 blender 
               
               
                 Soy Bean 
                 reducing 
               
               
                 Leaves 
                 with water 
               
               
                   
               
            
           
         
       
     
     Example X 
     Evaluated mixtures of alfalfa powder of 20 grain per gallon and 20 grain and 4 grain sugar. Later in the day, the alfalfa/powder mixture resulted in zebra mussels open slightly but responsive to touch, while alfalfa powder only resulted in zebra mussels not open upon observation. See Table 13. 
     
       
         
           
               
               
               
             
               
                 TABLE 13 
               
               
                   
               
               
                   
                   
                 Time to 
               
               
                 Treatment 
                 Comments 
                 Death 
               
               
                   
               
             
            
               
                 20 grain Alfalfa Powder 
                 Put in mixture for 3 hours 
                 &lt;36 hours 
               
               
                 with 4 grain sugar 
                 then put in clear water 
               
               
                 (in 1-gallon water) 
               
               
                 20 grain Alfalfa Powder with 20 
                 Also killed algae 
                 &lt;24 hours 
               
               
                 grain sugar (in 1-gallon water) 
               
               
                   
               
            
           
         
       
     
     Example XI 
     Additional testing was performed, as described along with the results in Table 14, below. 
     
       
         
           
               
               
               
             
               
                 TABLE 14 
               
               
                   
               
               
                   
                   
                 Time to 
               
               
                 Treatment 
                 Comments 
                 Death 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                 ⅔ Pellet Alfalfa, 
                 Drop mixture of 32 ounce cup into lake water, 
                 &lt;7 
                 hours 
               
               
                 ⅙ Powder Alfalfa 
                 conditions windy, boating waves and shower 
               
               
                 and ⅙ Spirulina 
                 afterwards. Immediate area of Zebra Mussels 
               
               
                   
                 were non responsive after 7 hours 
               
               
                 Alfalfa Pellets 
                 Slow dissipation allowing fish to live, 
                 &lt;48 
                 hours 
               
               
                 (15 grain) 
                 because of slow change in Chlorophyll 
               
               
                   
                 Levels 
               
               
                 Alfalfa Powder or 
                 Place fish in when levels at 25 μg/L or above 
                 &lt;12 
                 hours 
               
               
                 Spirulina to 
                 resulted in quick death of gold fish. 
               
               
                 chlorophyll levels of 
               
               
                 35 μg/L to 45 μg/L 
               
               
                 Alfalfa Pellets to 
                 Gold fish and Zebra mussels in place from start 
                 &lt;36 
                 hours 
               
               
                 allow melting 
                 while melting pellets, chlorophyll levels above 
               
               
                   
                 30 μg/L reached. Fished lived several weeks 
               
               
                   
                 before removing still live. Fish were quicker to 
               
               
                   
                 respond after test was completed. Noticed that 
               
               
                   
                 fish ate the alfalfa product during test. 
               
               
                 Alfalfa Pellets 
                 Placed both fish and zebra mussels into mixture 
                 &lt;36 
                 hours 
               
               
                 dusted with 
                 at the beginning. Again, levels above 30 μg/L 
               
               
                 Spirulina 
                 reached, and fish lived in the environment for 
               
               
                   
                 several weeks until removed. 
               
               
                 Lake water having 
                 Two 5-gallon buckets were used, placing Zebra 
                 &lt;48 
                 hours 
               
               
                 high chlorophyll 
                 Mussels attached to rocks in each bucket. Initial 
               
               
                 concentration 
                 lake water added measured 25 μg/L chlorophyll, 
               
               
                   
                 12 hours later chlorophyll levels measured 9 
               
               
                   
                 μg/L average. Replaced water with lake water 
               
               
                   
                 measuring 20 μg/L, after 48 hours the 
               
               
                   
                 chlorophyll levels measured 7 μg/L average. 
               
               
                   
               
            
           
         
       
     
     Example XII 
     Controlled specimens of zebra mussels were placed in both lake water and distilled water with recirculation pumps. The group with lake water was replenished with lake water as evaporation occurred. Distilled water was replenished with distilled water as needed. Zebra mussels lasted approximately 4 weeks in the distilled water, while zebra mussels lived for 6 to 8 weeks in the lake water environment. It is believed that starvation occurred with the distilled water, while the replenishing of lake water provided some nutrients. 
     Example XIII 
     Additional testing was performed to study the effects on water and mussel kill of various treatments. 
     Kill testing was performed using 20 grain of various treatments in 2000 ml of distilled water. Table 15 shows the treatment type, the high/low chlorophyll concentration over 48 hours of testing (variation due to due to break down of material and cell rupture when hydrated), and the zebra mussel time to death (in hours). 
     
       
         
           
               
               
               
               
             
               
                 TABLE 15 
               
               
                   
               
               
                   
                   
                   
                 Time to 
               
               
                   
                 Low 
                 High 
                 Death 
               
               
                 Treatment 
                 Chlorophyll 
                 Chlorophyll 
                 (h) 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Ground Soy Bean Leaves 
                 44 
                 μg/L 
                 199 
                 μg/L 
                 24 
               
               
                 Ground Duck Defecation 
                 43 
                 μg/L 
                 70 
                 μg/L 
                 24 
               
               
                 Ground Chicken Defecation 
                 15 
                 μg/L 
                 36 
                 μg/L 
                 10 
               
               
                 Distilled Water Only 
                 1 
                 μg/L 
                 1 
                 μg/L 
                 No Death 
               
               
                   
                   
                   
                   
                   
                 3 Days Stopped 
               
               
                   
               
            
           
         
       
     
     Testing was performed to determine water conditions using 20 grain of various treatments in 2000 ml of distilled water. Table 16 shows the treatment type, the high/low chlorophyll concentration over 48 hours of testing (variation due to due to break down of material and cell rupture when hydrated), turbidity, and pH. 
     
       
         
           
               
               
               
               
               
             
               
                 TABLE 16 
               
               
                   
               
               
                   
                 Low 
                 High 
                   
                   
               
               
                 Treatment 
                 Chlorophyll 
                 Chlorophyll 
                 Turbidity 
                 pH 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Ground Soy Bean Leaves 
                 50 
                 μg/L 
                 111 
                 μg/L 
                 50 
                 7.9 
               
               
                 Bought Ground Alfalfa 
                 40 
                 μg/L 
                 92 
                 μg/L 
                 40 
                 7.9 
               
               
                 Ground Duck Defecation 
                 32 
                 μg/L 
                 49 
                 μg/L 
                 40 
                 7.8 
               
               
                 Ground Chicken Defecation 
                 10 
                 μg/L 
                 11 
                 μg/L 
                 50 
                 7.4 
               
               
                 Ground White Paper 
                 2 
                 μg/L 
                 4 
                 μg/L 
                 — 
                 — 
               
               
                   
               
            
           
         
       
     
     Kill testing was performed using differing grain weights of various treatments in 2000 ml of distilled water. Table 17 shows the treatment type, the high/low chlorophyll concentration over 48 hours of testing (variation depended on stirred or settled), chlorophyll flotation, pH, and the zebra mussel time to death (in hours). See Table 17. 
     
       
         
           
               
               
               
               
               
               
             
               
                 TABLE 17 
               
               
                   
               
               
                   
                 Chlorophyll 
                 Chlorophyll 
                   
                 Time to 
                   
               
               
                   
                 Not Stirred, 
                 Stirred, 
                 Chlorophyll 
                 Death 
               
               
                 Treatment 
                 μg/L 
                 μg/L 
                 Flotation 
                 (hrs) 
                 pH 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 5 grain Soy Bean Leaf 
                 10 
                 58 
                 17% 
                 Approx 72 
                 7.7 
               
               
                 Powder 
               
               
                 5 grain Soy Bean Leaf 
                 18 
                 64 
                 28% 
                 Approx 72 
                 7.6 
               
               
                 Powder w/5 Grain 
               
               
                 Dried Anchovy 
               
               
                 10 Grain Soy Bean 
                 24 
                 135 
                 18% 
                 &lt;48 
                 7.7 
               
               
                 Leaf Powder 
               
               
                 10 Grain Soy Bean 
                 28 
                 134 
                 21% 
                 &lt;39 
                 7.7 
               
               
                 Leaf Powder w/5 
               
               
                 Grain Dried Anchovy 
               
               
                 5 Grain Spirulina 
                 123 
                 132 
                 93% 
                 &lt;24 
                 7.7 
               
               
                 5 Grain Spirulina w/5 
                 76 
                 86 
                 88% 
                  &lt;6 
                 7.7 
               
               
                 Grain Dried Anchovy 
               
               
                 Distilled Water Only 
                 0 
                 1 
                   
                 Stopped 
                 7.7 
               
               
                   
                   
                   
                   
                 (Alive for 
               
               
                   
                   
                   
                   
                 Days) 
               
               
                   
               
            
           
         
       
     
     Testing was performed to determine the radiation benefit of chlorophyll being present in a body of water. As shown in Table 18, below, chlorophyll treatments increased the temperature of the water when exposed to sunlight. Higher levels of chlorophyll would not only absorb more energy in the winter, but also it provides more oxygen to the environment. 
     
       
         
           
               
               
               
             
               
                 TABLE 18 
               
               
                   
               
               
                   
                 Chlorophyll 
                 Temp. (° F.) 
               
               
                 Treatment 
                 μg/L 
                 Time 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Soy Bean Leaf 
                 15 
                 74 @ 
                 79 @ 
                 95 @ 
                 98 @ 
                 104 @ 
               
               
                 Powder 
                   
                 10:45am 
                 11am 
                 12pm 
                 1pm 
                 4pm 
               
               
                 Distilled Water 
                 0 
                 74 @ 
                 78 @ 
                 94 @ 
                 97 @ 
                 103 @ 
               
               
                   
                   
                 10:45am 
                 11am 
                 12pm 
                 1pm 
                 4pm 
               
               
                 Soy Bean Leaf 
                 &gt;199 
                 76 @ 
                 89 @ 
                 100 @ 
                 104 @ 
                 105 @ 
               
               
                 Powder 
                   
                 11am 
                 11am 
                 12pm 
                 1pm 
                 4pm 
               
               
                 Distilled Water 
                 0 
                 76 @ 
                 87 @ 
                 96 @ 
                 100 @ 
                 101 @ 
               
               
                   
                   
                 11am 
                 11am 
                 12pm 
                 1pm 
                 4pm 
               
               
                 Spirulina Powder 
                 103 
                 76 @ 
                 87 @ 
                 95 @ 
                 100 @ 
                 102 @ 
               
               
                   
                   
                 11am 
                 11am 
                 12pm 
                 1pm 
                 4pm 
               
               
                 Distilled Water 
                 0 
                 76 @ 
                 86 @ 
                 93 @ 
                 97 @ 
                 98 @ 
               
               
                   
                   
                 11am 
                 11am 
                 12pm 
                 1pm 
                 4pm 
               
               
                   
               
            
           
         
       
     
     Example XIV 
     During the above testing described in Examples I-XIII, additional observations were noted and are summarized below:
         Although chlorella is a more expensive treatment, it generally exhibited better acceptance in water testing.   Wheat grass and barley grass settle very quickly, so difficult to keep suspended.   Soy bean leaf powder tended to be more effective than alfalfa, which tended to be more effective than grasses.   Spirulina tended to be more effective than all of the above, particularly for closed systems, but may be too expensive for large bodies of water.   All treatment intake can be enhance by flavoring with tasteful food sources.   Liquid treatments were preferable over powder, which were preferable over pellets, for administering.   Gold fish, insects, and spiral snails lived as zebra mussels dies in same treated water (when the chlorophyll levels were gradually raised so as not to shock fish).       

     Example XV 
     Fresh water moss was collected from a lake infested with zebra mussels, and the moss was surrounding, floating above, and touching the zebra mussel colonies. The fresh water moss was collected and processed in a blender, similar to what was done with leaves in the examples above. Three treatment samples were prepared, with one flavored with anchovy. The three samples were added to separate bodies of water at approximately 7:00 pm. This resulted in chlorophyll readings greater than an estimated 199 μg/L at 8:00 pm. By 5:00 am the following morning, there were no responses by the open zebra mussels. Additionally, the chlorophyll level readings were 199 μg/L, 144 μg/L and 113 μg/L; it seemed that the number of Zebra Mussels within each container determine the final chlorophyll levels. Notably, spiral snails collected survived testing. 
     Example XVI 
     In view of the studies above, further investigation and observations were performed to better understand the mechanisms of mussel kill and control. 
     Observations 
     It was observed that  Dreissenid  mussel infestations were found in chlorophyll-a containing Oligotrophic and Mesotrophic areas of lakes, but not observed in Eutrophic through Hypereutrophic levels. It is believed that the  Dreissenid  mussels are unable to control their eating habits, causing suffocation/blockage in waters that have not been systematically filtered. This action is observed in partially infested lakes, where in areas with an inlet of clean water run-off zebra mussels were found to be plentiful, but in other areas no zebra mussels were found on docks or equipment where chlorophyll-a measured above 30 mg/L. 
     Discussion 
     Based on the observations above, it is believed that by increasing the aroma and taste of oligotrophic and mesotrophic waters where infestations of  Dreissenid  mussels can be found, the  Dreissenid  mussels are unable to control their eating habits, similar to many other animals that are provided food having strong, favorable taste or aroma. For example, this is seen with cattle. When allowed to over-consume feed, cattle will become bloated, a condition which is usually precipitated by the rapid consumption of lush legume pasture species and can lead to death. It is believed that  Dreissenid  mussels react to the same lush legume aroma and taste, as seen with soybean and clover foliage, which can be measured by the concentration of chlorophyll-a within the water. Notably, however, no other aquatic animals were observed to over-consume in the high chlorophyll-a containing eutrophic-hypertrophic conditions where the  Dreissenid  mussels were observed to react. 
     Testing 
     The observations were tested by first adding about a teaspoon of ground-up dried anchovies to a quart of water and allowing to sour, becoming very distasteful to the smell. Chlorophyll-a was added to the soured anchovy mixture in an amount sufficient to raise the concentration of chlorophyll in the subsequently treated water (see below) to about 30 μg/L. The mixture was added to a 10-gallon aquarium container holding water and a sample of approximately 40 live zebra mussels. Addition of the mixture resulted in closure of the zebra mussels. After four days, the soured anchovy mixture was removed and replaced with distilled water mixed with a processed plant product. The processed plant product raised the chlorophyll-a level in the distilled water, which acquired a more favorable aroma, resulting in opening of the zebra mussels and filtration feeding to begin. This led the mussels irresistibly over eating. This test confirms that the chlorophyll-a aroma and taste is more favorable than other products to the mussels, which leads to overeating and eventual death. 
     Plant Processing 
     In view of the testing and observations above, certain considerations should be made in the processing of plants to release the aroma and taste while retaining its fibers and/or oils. For example, processed plant particles (e.g., powder particles) should preferably be in a measurable range of about 0.05 to about 650 micrometers in all directions, more preferably ranging from about 0.2 to about 400 micrometers maximum in any direction to allow normal ingestion by the mussels. The size can be determined at the time of application or after dissolving or breaking up when in contact with a liquid or water environment that is introduced into water bodies where  Dreissenid  mussels inhabit. 
     Example XVII 
     Efficacy testing was conducted at chlorophyll concentrations above 200 μg/L. However, because the Chlorophyll Meter has an upper measurable limit of about 200 μg/L, higher concentrations were estimated by extrapolation. 
     As shown in Table 19 below, grains of alfalfa powder (ground powder) and spirulina powder (finely ground powder) were added to distilled water two at a time and mixed, and chlorophyll concentration was measured. When the results were plotted, a general trendline was observed having an R 2  value of greater than 0.9 (See  FIG. 1  and  FIG. 2 ). Thus, the trendline can be extrapolated beyond the measurable limit of 200 μg/L to accurately estimate higher concentrations based on the number of powder grains added to the water (See  FIG. 3  and  FIG. 4 ). 
     
       
         
           
               
               
               
             
               
                 TABLE 19 
               
             
            
               
                   
               
               
                 Example: ALFALFA POWDER 
                   
                 Example: SPIRULINA POWDER 
               
            
           
           
               
               
               
               
               
            
               
                   
                 CHLOROPHYLL 
                 DISTILLED WATER 
                   
                 CHLOROPHYLL 
               
               
                 GRAINS 
                 ug/L 
                 GRAINS 
                 GRAINS 
                 ug/L 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 2 
                 15 
                 56,000 
                 2 
                 36 
               
               
                 4 
                 25 
                 56,000 
                 6 
                 77 
               
               
                 6 
                 37 
                 56,000 
                 8 
                 91 
               
               
                 8 
                 55 
                 56,000 
                 12 
                 129 
               
               
                 10 
                 56 
                 56,000 
                 16 
                 147 
               
               
                 12 
                 70 
                 56,000 
                 18 
                 158 
               
               
                 14 
                 79 
                 56,000 
                 20 
                 167 
               
               
                 16 
                 91 
                 56,000 
                 24 
                 177 
               
               
                 18 
                 100 
                 56,000 
                 28 
                 177 
               
               
                 20 
                 107 
                 56,000 
                 30 
                 198 
               
               
                 22 
                 123 
                 56,000 
               
               
                 24 
                 127 
                 56,000 
               
               
                 26 
                 137 
                 56,000 
               
               
                 28 
                 149 
                 56,000 
               
               
                 30 
                 152 
                 56,000 
               
               
                 32 
                 150 
                 56,000 
               
               
                 34 
                 163 
                 56,000 
               
               
                 36 
                 162 
                 56,000 
               
               
                 38 
                 170 
                 56,000 
               
               
                 40 
                 188 
                 56,000 
               
               
                 42 
                 191 
                 56,000 
               
               
                 44 
                 199 
                 56,000 
               
               
                   
               
            
           
         
       
     
     To determine that consistent mussel eradication could be achieved at higher chlorophyll levels (above 200 μg/L) and different powder consistencies, two tests were performed at concentrations of 52,000 μg/L (prepared using spirulina powder) and 160,000 μg/L (prepared using alfalfa powder). The chlorophyll concentrations were prepared in buckets by adding the appropriate amount of spirulina powder or alfalfa powder to buckets of distilled water based on the extrapolated trendlines described above (See  FIG. 3  and  FIG. 4 ). 
     Live zebra mussels attached to a rock were introduced into the respective buckets containing about 52,000 μg/L (spirulina) and 160,000 μg/L (alfalfa), prepared as described above. In both instances, the rocks holding the zebra mussels were removed after 4 hours, and the zebra mussels were not responsive. Additionally, the zebra mussels that were killed using spirulina at 52,000 μg/L showed a green tint upon visual examination, evidencing spirulina ingestion.