Patent Publication Number: US-2020299164-A1

Title: Method for water remediation and desalination via selectively breeding and cultivating generations of microbes

Description:
This application is related to Applications ______ entitled “MICROBIAL SOIL ENHANCEMENTS” and ______ entitled “ANIMAL FEED STOCK USING MICROBIAL ENHANCEMENTS”, all of which are filed concurrently herewith, and the contents of which are incorporated-by-reference. 
     BACKGROUND 
     The present invention relates to water remediation. 
     For aquatic applications, algae blooms are one of the most common problems encountered by lake and pond owners. More than just aesthetically unappealing, certain forms of algae may harbor potentially toxic bacteria. Conventional ways of solving algae blooms are expensive and/or environmentally harmful as follows: 
     Mechanical intervention—Mechanical methods of algae control include removing blooms with a rake, seine, screen or other tool. While useful for quickly remediating harmful or invasive growths, mechanical controls do not address the underlying conditions that caused the problem. To address the underlying conditions, they must be used in conjunction with other methods. 
     Physical intervention—Some lakes and ponds are more prone to algae growth than others—factors such as aeration, light and nutrient content all affect whether or not algae will bloom. Adding an aerator or making other changes that promote these positive physical characteristics is one method of preventing algae outbreaks. 
     Biological intervention—Ponds and lakes are complex ecosystems. Under ideal conditions, they will attain a balance in which invasive algae growth is kept to a minimum. By introducing certain fish or insects into this ecosystem, we can help it attain a state of equilibrium that minimizes algae blooms naturally. 
     Material intervention—One of the most common methods utilized for controlling noxious algal blooms is with the use of algaecides. When utilized by a professional, algaecides can provide safe and effective management of harmful algae blooms. Out of balance populations of algae can be quickly brought under control when applications are conducted in a timely manner and under appropriate environmental conditions. Fish populations can benefit as well from improved spawning habitat and increased access to forage. 
     SUMMARY 
     In one aspect, a method for water remediation includes preparing a microbial solution with microbes, a growth medium, and water; iteratively and selectively breeding generations of microbes to arrive at a predetermined microbial solution in a highly concentrated form of at least 1×10 9  cfu/ml (colony-forming units per milliliter); and dispensing the microbial solution into the water, wherein the microbes metabolizes excess nutrients and uses microbial desalination to reduce water salinity. 
     Implementations may include one or more of the following. A member of  Bacillus  can be the microbe. The growth medium can be a carbon source. The growth medium can be sugar, molasses, or maltodextrin. The solution can have 1 part microbes, 10 part carbon source, and 1000 parts water. The solution can use as a carrier one of: amino acid, maltodextrin, wheat, or corn. The microbe can be one of:  Bacillus  ( B. )  acidiceler, B. acidicola, B. acidiproducens, B. acidocaldarius, B. acidoterrestrisr, B. aeolius, B. aerius, B. aerophilus, B. agaradhaerens, B. agri, B. aidingensis, B. akibai, B. alcalophilus, B. algicola, B. alginolyticus, B. alkalidiazotrophicus, B. alkalinitrilicus, B. alkalisediminis, B. alkalitelluris, B. altitudinis, B. alveayuensis, B. alvei, B. amyloliquefaciens, B. a.  subsp.  amyloliquefaciens, B. a.  subsp.  plantarum, B. amylolyticus, B. andreesenii, B. aneurinilyticus, B. anthracia, B. aquimaris, B. arenosi, B. arseniciselenatis, B. arsenicus, B. aurantiacus, B. arvi, B. aryabhattai, B. asahii, B. atrophaeus, B. axarquiensis, B. azotofixans, B. azotoformans, B. badius, B. barbaricus, B. bataviensis, B. beijingensis, B. benzoevorans, B. beringensis, B. berkeleyi, B. beveridgei, B. bogoriensis, B. boroniphilus, B. borstelensis, B. brevis migula, B. butanolivorans, B. canaveralius, B. carboniphilus, B. cecembensis, B. cellulosilyticus, B. centrosporus, B. cereus, B. chagannorensis, B. chitinolyticus, B. chondroitinus, B. choshinensis, B. chungangensis, B. cibi, B. circulans, B. clarkii, B. clausii, B. coagulans, B. coahuilensis, B. cohnii, B. composti, B. curdlanolyticus, B. cycloheptanicus, B. cytotoxicus, B. daliensis, B. decisifrondis, B. decolorationis, B. deserti, B. dipsosauri, B. drentensis, B. edaphicus, B. ehimensis, B. eiseniae, B. enclensis, B. endophyticus, B. endoradicis, B. farraginis, B. fastidiosus, B. fengqiuensis, B. firmus, B. flexus, B. foraminis, B. fordii, B. formosus, B. fortis, B. fumarioli, B. funiculus, B. fusiformis, B. galactophilus, B. galactosidilyticus, B. galliciensis, B. gelatini, B. gibsonii, B. ginsengi, B. ginsengihumi, B. ginsengisoli, B. globisporus, B. g.  subsp.  globisporus, B. g.  subsp.  marinus, B. glucanolyticus, B. gordonae, B. gottheilii, B. graminis, B. halmapalus, B. haloalkaliphilus, B. halochares, B. halodenitrificans, B. halodurans, B. halophilus, B. halosaccharovorans, B. hemicellulosilyticus, B. hemicentroti, B. herbersteinensis, B. horikoshii, B. horneckiae, B. horti, B. huizhouensis, B. humi, B. hwajinpoensis, B. idriensis, B. indicus, B. infantis, B. infernus, B. insolitus, B. invictae, B. iranensis, B. isabeliae, B. isronensis, B. jeotgali, B. kaustophilus, B. kobensis, B. kochii, B. kokeshiiformis, B. koreensis, B. korlensis, B. kribbensis, B. krulwichiae, B. laevolacticus, B. larvae, B. laterosporus, B. lautus, B. lehensis, B. lentimorbus, B. lentus, B. licheniformis, B. ligniniphilus, B. litoralis, B. locisalis, B. luciferensis, B. luteolus, B. luteus, B. macauensis, B. macerans, B. macquariensis, B. macyae, B. malacitensis, B. mannanilyticus, B. marisflavi, B. marismortui, B. marmarensis, B. massiliensis, B. mesonae, B. methanolicus, B. methylotrophicus, B. migulanus, B. mojavensis, B. mucilaginosus, B. muralis, B. murimartini, B. mycoides, B. naganoensis, B. nanhaiensis, B. nanhaiisediminis, B. nealsonii, B. neidei, B. neizhouensis, B. niabensis, B. niacini, B. novalis, B. oceanisediminis, B. odysseyi, B. okhensis, B. okuhidensis, B. oleronius, B. oryzaecorticis, B. oshimensis, B. pabuli, B. pakistanensis, B. pallidus, B. pallidus, B. panacisoli, B. panaciterrae, B. pantothenticus, B. parabrevis, B. paraflexus, B. pasteurii, B. patagoniensis, B. peoriae, B. persepolensis, B. persicus, B. pervagus, B. plakortidis, B. pocheonensis, B. polygoni, B. polymyxa, B. popilliae, B. pseudalcalophilus, B. pseudofirmus, B. pseudomycoides, B. psychrodurans, B. psychrophilus, B. psychrosaccharolyticus, B. psychrotolerans, B. pulvifaciens, B. pumilus, B. purgationiresistens, B. pycnus, B. qingdaonensis, B. qingshengii, B. reuszeri, B. rhizosphaerae, B. rigui, B. ruris, B. safensis, B. salarius, B. salexigens, B. saliphilus, B. schlegelii, B. sediminis, B. selenatarsenatis, B. selenitireducens, B. seohaeanensis, B. shacheensis, B. shackletonii, B. siamensis, B. silvestris, B. simplex, B. siralis, B. smithii, B. soli, B. solimangrovi, B. solisalsi, B. songklensis, B. sonorensis, B. sphaericus, B. sporothermodurans, B. stearothermophilus, B. stratosphericus, B. subterraneus, B. subtilis, B. s.  subsp.  inaquosorum, B. s.  subsp.  spizizenii, B. s.  subsp.  subtilis, B. taeanensis, B. tequilensis, B. thermantarcticus, B. thermoaerophilus, B. thermoamylovorans, B. thermocatenulatus, B. thermocloacae, B. thermocopriae, B. thermodenitrificans, B. thermoglucosidasius, B. thermolactis, B. thermoleovorans, B. thermophilus, B. thermoruber, B. thermosphaericus, B. thiaminolyticus, B. thioparans, B. thuringiensis, B. tianshenii, B. trypoxylicola, B. tusciae, B. validus, B. vallismortis, B. vedderi, B. velezensis, B. vietnamensis, B. vireti, B. vulcani, B. wakoensis, B. weihenstephanensis, B. xiamenensis, B. xiaoxiensis,  and  B. zhanjiangensis.    
     Advantages of the above systems may include one or more of the following. The selectively bred bacteria form a bio-solution to help maintain ponds and lakes clean and clear the natural way. The microbes to break out the excess nutrients, pull them away from the water column and restate the water to its naturally beautiful and healthy condition without the undesirable side effects of chemicals. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The novel features that are considered characteristic of the present invention are set forth with particularity in the appended claims. The invention itself, however, both as to its organization and its method of operation, together with additional objects and advantages thereof, will best be understood from the following description of specific embodiments when read in connection with the accompanying drawings, in which: 
         FIG. 1  shows an exemplary process to selectively breed the microbes for aquaculture or water use. 
         FIG. 2  shows an exemplary process to produce the microbial products. 
         FIGS. 3A-3B  show exemplary microbial activity expressed by different  Bacillus  spp. strains. 
     
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     The following detailed description is of the best presently contemplated modes of carrying out the invention. This description is not to be taken in a limiting sense, but is made merely for the purpose of illustrating the general principles of the invention, since the scope of the invention best is defined by the appended claims. 
     Structural and operational characteristics attributed to forms of the invention at first described also shall be attributed to forms later described, unless such characteristics obviously are inapplicable, or unless specific exception is made. 
     A selectively bred microbial solution is disclosed with multiple single microbial series separately cultivated and followed with cross cultivation among those microbial series in a specific sequence and contains each of those microbial series, and by-products produced by those crossly cultivated microbial series are used for applications in modifying water quality. After the selective breeding through the fermentation, the selectively bred naturally-occurring microorganism is provided as a highly concentrated solution with the ability to digest excess nutrients to clear the water. 
     In a water enhancement embodiment, the selectively bred bacteria forms a bio-solution to help maintain ponds and lakes clean and clear the natural way. The microbes to break out the excess nutrients, pull them away from the water column and restate the water to its naturally beautiful and healthy condition without the undesirable side effects of chemicals. 
       FIG. 1  shows an exemplary process to selectively breed the microbes for water use. First, fermentation media are prepared with a nutrient supply (1). The nutrients can include a carbon source Dextrose or Glucose. Additional carbon sources can be used with the dextrose or glucose singly or in combination. For example, another carbon source can be sucrose, for example. Next, a nitrogen source is provide such as soy protein that has not been genetically modified (2). Next, in (3), micronutrients—Calcium, Magnesium and Zinc are provided. A person of ordinary skilled in the art appreciates that various compositions of the fermentation media can be prepared so long as the nutrients, one or more of the carbon sources, and the micronutrients are included. 
     In (4), the fermentation media is prepared using water supply and sterilized using stream sterilizer at 120 degrees Celsius for 45 minutes, but the temperature and time can be varied in accordance with tank volume. In (5), the process produces the microbial products, as is detailed in  FIG. 2 . At each stage, quality control methods are applied using standard plate count method for  Shigella, E. Coli, Salmonella Yersinia  and  Psuedomonas beroginosa  for their absence. All products are manufactured according to USEPA (United States Environmental Protection Agency) standards. 
     The Microbial Strain selection and profile of microbial genes are carefully selected to form the formulation of products. Through strain selections, screening and improvement, the system generates various microbes for clearing water. For example,  Bacillus subtilus  has 4,100 genes. These genes each contain approximately 2000 traits. Each one of these traits and its mutation has over 1000 profile and sub-profile. 
     The biodiversity of  Bacillus  group and beneficial traits of  Bacillus  species are useful in removing excess nutrients from the water column.  Bacillus  genus is widely spread in nature.  Bacillus  species such as  B. subtilus, B. amyloliquefaciens, B. lichniformis, Bacillus amyloliquefaciens, Bacillus lichniformis, Bacillus pumilus, Bacillus subtilis,  or  Pseudonomas putida  are carefully selected, for their specific profile which contains beneficial traits. 
     Each single microbial series is separately cultivated in its designated cultivation medium, and the optimal pH in the growing and reproduction of different microbial series also varies. Therefore, proper control and regulation of pH of the cultivation medium are provided in the course of bacterial cultivation and fermentation. The microbial series acquires energy through aerobic respiration. However, the aerobic respiration generally has to rely upon only the oxygen dissolved in the cultivation medium, i.e., the dissolved oxygen, and the containment of the dissolved oxygen in the cultivation medium is not always provided in sufficient amount and will be soonest consumed by bacteria since oxygen is difficult to get dissolved in water. Therefore, constant air supply to the microbial series is provided without interruption in the course of the cultivation and fermentation of the microbial series. Compositions of cultivation medium selected and the optimal growing environment conditions for each microbial series are detailed as follows: 
       FIG. 3A-3B  shows exemplary microbial activity expressed by different  Bacillus  spp. Strains.  FIG. 3A  shows exemplary  Bacillus  spp. antagonistic activity against  Fusarium solani ; while  FIG. 3B  shows exemplary fungal cell wall degradation, cell lysis and cytoplasm bleeding due to  Bacillus  spp. extracellular enzymes. 
     In a water cleaning embodiment called BIOPOND, the solution provides an all-natural, extremely concentrated beneficial microbes. When applied to water, the solution reduces ammonia, nitrate, nitrite and phosphorus. The solution binds phosphate, reduces water salinity, COD, and BOD. The solution removes toxic blue-green algae, hydrogen sulfide, and odors, reduces organic waste, bottom sludge and suspended solids, is safe for fish, pets, birds, livestock, plants and wildlife. The solution helps maintain optimum pH and create a balanced ecosystem and is highly effective biological filtration process of mineralization, nitrification and denitrification. Water is cleared naturally by removing excess nutrients from the pond. The BioPond application is simple and costs efficient solution available either by the biodegradable packets ready to toss or easy to incubate for industrial cost effective applications. The dosage is calculated based on the volume of water in acre-foot. 
     BIOPOND is a blend of naturally occurring beneficial aerobic and facultative anaerobic microbes that is effective in reducing water column nutrients in lakes, ponds, and organic wastewater. BioPond improves water quality without the undesirable side effects of chemical treatments. This is done as BioPond rapidly metabolizes excess nutrients and pulling them from the water column and uses microbial desalination cells to reduce water salinity. 
     The application of beneficial microbes is the key to a healthy water environment, effortless, all-natural, eco-friendly simple solution to manage ponds, lakes and wastewater treatment in a balanced ecosystem. Beneficial microbes are “good” bacteria, naturally occurring microorganisms that perform their work by producing enzymes and provide intelligent to detect the organics present to continue produce specifically those enzymes needed to break down the organics present. These microbes are capable of improving water quality and clarity by targeting organic particulates and excess nutrients, such as Nitrogen and Phosphorus, brought in by various sources. Since they are naturally occurring and present in low concentrations, microbes in the ponds are not able to affecting the water quality and clarity on their own. The BioPond application of concentrated beneficial microbes can exponentially boost their populations giving them the ability to affect water quality and clarity. 
     Any microbial spores and/or colonies can be preserved using methods and solutions of some embodiments. Spores and/or colonies of beneficial water and water pathogen biological control microorganisms are preferred. Microorganisms that grow rapidly and colonize substrata in water after treatment with compositions of the invention are particularly preferred. These include, but are not limited to bacteria, e.g.,  Bacillus  species such as  Bacillus subtilis, Bacillus cereus, Bacillus penetrans, Bacillus licheniformis;  fungi, e.g.,  Trichoderma  species such as  Trichoderma hamatum, Trichoderma harzianum, Trichoderma polysporum, Trichoderma konigii,  and  Trichoderma viride;  and yeast species such as  Saccharomyces cerevisiae . As illustrated below, mixtures of microorganisms can also be preserved, and are preferred in many embodiments. Examples are given hereafter. 
     Water miscible dry powders and/or granules such as lyophilized preparations of spores and/or colonies are preferred in many embodiments. The amount of spores or microorganisms added to solutions of the invention is not fixed per se, and necessarily is dependent upon the degree of water remediation required, the number and identity of microorganism species needed in the formulation, and the concentration of other ingredients in the formulation. Typical embodiments contain sufficient spores and/or colonies to deliver from about 1000 to about 1,000,000 colony forming units (CFU) per square foot when the preparation is delivered. 
     The carrier for the microbes solution can be one of: maltodextrin, wheat, or corn. The resulting solution has a high concentration of microbes as detailed in the following examples. The colony-forming unit (CFU or cfu) is a measure of viable bacterial or fungal cells. CFU measures only viable cells. For convenience the results are given as CFU/mL (colony-forming units per milliliter) for liquids, and CFU/g (colony-forming units per gram) for solids. 
     EXAMPLE 1 (BIOPOND) 
     Microbes: 
     
         
         
           
               Bacillus amyloliquefaciens  at 0.91×10{circumflex over ( )}9 cfu/g 
               Bacillus lichniformis  at 0.30×10{circumflex over ( )}9 cfu/g 
               Bacillus pumilus  at 0.71×10{circumflex over ( )}9 cfu/g 
               Bacillus subtilis  at 1.10×10{circumflex over ( )}9 cfu/g 
           
         
       
    
     Carrier: Wheat 
     EXAMPLE 2 (BIOPOND) 
     Microbes: 
     
         
         
           
               Bacillus amyloliquefaciens  at 4.45×10{circumflex over ( )}9 cfu/g 
               Bacillus lichniformis  at 1.48×10{circumflex over ( )}9 cfu/g 
               Bacillus pumilus  at 3.35×10{circumflex over ( )}9 cfu/g 
               Bacillus subtilis  at 4.75×10{circumflex over ( )}9 cfu/g 
               Pseudonomas putida  at 2.45×10{circumflex over ( )}8 cfu/g
 
Carrier: Corn Distiller and Dry Grain with Soluble
 
           
         
       
    
     EXAMPLE 3 (BIOPOND) 
     Microbes: 
     
         
         
           
               Bacillus amyloliquefaciens  at 4.45×10{circumflex over ( )}9 cfu/g 
               Bacillus lichniformis  at 1.48×10{circumflex over ( )}9 cfu/g 
               Bacillus pumilus  at 3.35×10{circumflex over ( )}9 cfu/g 
               Bacillus subtilis  at 4.75×10{circumflex over ( )}9 cfu/g 
               Pseudonomas putida  at 2.45×10{circumflex over ( )}8 cfu/g 
           
         
       
    
     Carrier: Maltodextrin and Amino Acid Complex 
     EXAMPLE 4 (BIOPOND) 
     Microbes: 
     
         
         
           
               Bacillus amyloliquefaciens  at 0.91×10{circumflex over ( )}9 cfu/g 
               Bacillus lichniformis  at 0.30×10{circumflex over ( )}9 cfu/g 
               Bacillus pumilus  at 0.71×10{circumflex over ( )}9 cfu/g 
               Bacillus subtilis  at 1.10×10{circumflex over ( )}9 cfu/g 
           
         
       
    
     Carrier: Wheat 
     EXAMPLE 5 (BIOPOND) 
     Microbes: 
     
         
         
           
               Bacillus amyloliquefaciens  at 1.30×10{circumflex over ( )}9 cfu/g 
               Bacillus lichniformis  at 0.30×10{circumflex over ( )}9 cfu/g 
               Bacillus pumilus  at 1.80×10{circumflex over ( )}9 cfu/g 
               Bacillus subtilis  at 1.80×10{circumflex over ( )}9 cfu/g 
           
         
       
    
     Carrier: Wheat 
     EXAMPLE 6 (BIOPOND) 
     Microbes: 
     
         
         
           
               Bacillus amyloliquefaciens  at 0.36×10{circumflex over ( )}9 cfu/g 
               Bacillus lichniformis  at 0.71×10{circumflex over ( )}9 cfu/g 
               Bacillus pumilus  at 0.50×10{circumflex over ( )}9 cfu/g 
               Bacillus subtilis  at 0.50×10{circumflex over ( )}9 cfu/g 
           
         
       
    
     Carrier: Distillers Dry Grain 
     EXAMPLE 7 (BIOPOND) 
     Microbes: 
     
         
         
           
               Bacillus amyloliquefaciens  at 4.45×10{circumflex over ( )}9 cfu/g 
               Bacillus lichniformis  at 1.48×10{circumflex over ( )}9 cfu/g 
               Bacillus pumilus  at 3.35×10{circumflex over ( )}9 cfu/g 
               Bacillus subtilis  at 4.75×10{circumflex over ( )}9 cfu/g 
               Pseudonomas putida  at 2.45×10{circumflex over ( )}8 cfu/g 
           
         
       
    
     Carrier: Maltodextrin and Amino Acid Complex 
       Bacillus  is a genus of gram-positive, rod-shaped bacteria and a member of the phylum Firmicutes.  Bacillus  species can be obligate aerobes (oxygen reliant), or facultative anaerobes (having the ability to be aerobic or anaerobic). They will test positive for the enzyme catalase when there has been oxygen used or present. Ubiquitous in nature,  Bacillus  includes both free-living (nonparasitic) and parasitic pathogenic species. Under stressful environmental conditions, the bacteria can produce oval endospores that are not true ‘spores’, but to which the bacteria can reduce themselves and remain in a dormant state for very long periods. These characteristics originally defined the genus, but not all such species are closely related, and many have been moved to other genera of the Firmicutes. Many species of  Bacillus  can produce copious amounts of enzymes which are made use of in different industries. Some species can form intracellular inclusions of polyhydroxyalkanoates under certain adverse environmental conditions, as in a lack of elements such as phosphorus, nitrogen, or oxygen combined with an excessive supply of carbon sources. Many  Bacillus  species are able to secrete large quantities of enzymes.  Bacillus amyloliquefaciens  is the source of a natural antibiotic protein barnase (a ribonuclease), alpha amylase used in starch hydrolysis, the protease subtilisin used with detergents, and the BamH1 restriction enzyme used in DNA research. 
       Bacillus  ( B. )  subtilis  is a Gram-positive bacterium, rod-shaped and catalase-positive.  B. subtilis  cells are typically rod-shaped, and are about 4-10 micrometers (μm) long and 0.25-1.0 μm in diameter, with a cell volume of about 4.6 fL at stationary phase. As with other members of the genus  Bacillus,  it can form an endospore, to survive extreme environmental conditions of temperature and desiccation.  B. subtilis  is heavily flagellated, which gives it the ability to move quickly in liquids.  B. subtilis  has proven highly amenable to genetic manipulation, and has become widely adopted as a model organism for laboratory studies, especially of sporulation, which is a simplified example of cellular differentiation. In terms of popularity as a laboratory model organism,  B. subtilis  is often considered as the Gram-positive equivalent of  Escherichia coli,  an extensively studied Gram-negative bacterium. Other members of the  Bacillus  species can be used, including:
           Bacillus  ( B. )  acidiceler        B. acidicola        B. acidiproducens        B. acidocaldarius        B. acidoterrestris        B. aeolius        B. aerius        B. aerophilus        B. agaradhaerens        B. agri        B. aidingensis        B. akibai        B. alcalophilus        B. algicola        B. alginolyticus        B. alkalidiazotrophicus        B. alkalinitrilicus        B. alkalisediminis        B. alkalitelluris        B. altitudinis        B. alveayuensis        B. alvei        B. amyloliquefaciens        B. a.  subsp.  amyloliquefaciens        B. a.  subsp.  plantarum        B. amylolyticus        B. andreesenii        B. aneurinilyticus        B. anthracia        B. aquimaris        B. arenosi        B. arseniciselenatis        B. arsenicus        B. aurantiacus        B. arvi        B. aryabhattai        B. asahii        B. atrophaeus        B. axarquiensis        B. azotofixans        B. azotoformans        B. badius        B. barbaricus        B. bataviensis        B. beijingensis        B. benzoevorans        B. beringensis        B. berkeleyi        B. beveridgei        B. bogoriensis        B. boroniphilus        B. borstelensis        B. brevis migula        B. butanolivorans        B. canaveralius        B. carboniphilus        B. cecembensis        B. cellulosilyticus        B. centrosporus        B. cereus        B. chagannorensis        B. chitinolyticus        B. chondroitinus        B. choshinensis        B. chungangensis        B. cibi        B. circulans        B. clarkii        B. clausii        B. coagulans        B. coahuilensis        B. cohnii        B. composti        B. curdlanolyticus        B. cycloheptanicus        B. cytotoxicus        B. daliensis        B. decisifrondis        B. decolorationis        B. deserti        B. dipsosauri        B. drentensis        B. edaphicus        B. ehimensis        B. eiseniae        B. enclensis        B. endophyticus        B. endoradicis        B. farraginis        B. fastidiosus        B. fengqiuensis        B. firmus        B. flexus        B. foraminis        B. fordii        B. formosus        B. fortis        B. fumarioli        B. funiculus        B. fusiformis        B. galactophilus        B. galactosidilyticus        B. galliciensis        B. gelatini        B. gibsonii        B. ginsengi        B. ginsengihumi        B. ginsengisoli        B. globisporus        B. g.  subsp.  globisporus        B. g.  subsp.  marinus        B. glucanolyticus        B. gordonae        B. gottheilii        B. graminis        B. halmapalus        B. haloalkaliphilus        B. halochares        B. halodenitrificans        B. halodurans        B. halophilus        B. halosaccharovorans        B. hemicellulosilyticus        B. hemicentroti        B. herbersteinensis        B. horikoshii        B. horneckiae        B. horti        B. huizhouensis        B. humi        B. hwajinpoensis        B. idriensis        B. indicus        B. infantis        B. infernus        B. insolitus        B. invictae        B. iranensis        B. isabeliae        B. isronensis        B. jeotgali        B. kaustophilus        B. kobensis        B. kochii        B. kokeshiiformis        B. koreensis        B. korlensis        B. kribbensis        B. krulwichiae        B. laevolacticus        B. larvae        B. laterosporus        B. lautus        B. lehensis        B. lentimorbus        B. lentus        B. licheniformis        B. ligniniphilus        B. litoralis        B. locisalis        B. luciferensis        B. luteolus        B. luteus        B. macauensis        B. macerans        B. macquariensis        B. macyae        B. malacitensis        B. mannanilyticus        B. marisflavi        B. marismortui        B. marmarensis        B. massiliensis        B. mesonae        B. methanolicus        B. methylotrophicus        B. migulanus        B. mojavensis        B. mucilaginosus        B. muralis        B. murimartini        B. mycoides        B. naganoensis        B. nanhaiensis        B. nanhaiisediminis        B. nealsonii        B. neidei        B. neizhouensis        B. niabensis        B. niacini        B. novalis        B. oceanisediminis        B. odysseyi        B. okhensis        B. okuhidensis        B. oleronius        B. oryzaecorticis        B. oshimensis        B. pabuli        B. pakistanensis        B. pallidus        B. pallidus        B. panacisoli        B. panaciterrae        B. pantothenticus        B. parabrevis        B. paraflexus        B. pasteurii        B. patagoniensis        B. peoriae        B. persepolensis        B. persicus        B. pervagus        B. plakortidis        B. pocheonensis        B. polygoni        B. polymyxa        B. popilliae        B. pseudalcalophilus        B. pseudofirmus        B. pseudomycoides        B. psychrodurans        B. psychrophilus        B. psychrosaccharolyticus        B. psychrotolerans        B. pulvifaciens        B. pumilus        B. purgationiresistens        B. pycnus        B. qingdaonensis        B. qingshengii        B. reuszeri        B. rhizosphaerae        B. rigui        B. ruris        B. safensis        B. salarius        B. salexigens        B. saliphilus        B. schlegelii        B. sediminis        B. selenatarsenatis        B. selenitireducens        B. seohaeanensis        B. shacheensis        B. shackletonii        B. siamensis        B. silvestris        B. simplex        B. siralis        B. smithii        B. soli        B. solimangrovi        B. solisalsi        B. songklensis        B. sonorensis        B. sphaericus        B. sporothermodurans        B. stearothermophilus        B. stratosphericus        B. subterraneus        B. subtilis        B. s.  subsp.  inaquosorum        B. s.  subsp.  spizizenii        B. s.  subsp.  subtilis        B. taeanensis        B. tequilensis        B. thermantarcticus        B. thermoaerophilus        B. thermoamylovorans        B. thermocatenulatus        B. thermocloacae        B. thermocopriae        B. thermodenitrificans        B. thermoglucosidasius        B. thermolactis        B. thermoleovorans        B. thermophilus        B. thermoruber        B. thermosphaericus        B. thiaminolyticus        B. thioparans        B. thuringiensis        B. tianshenii        B. trypoxylicola        B. tusciae        B. validus        B. vallismortis        B. vedderi        B. velezensis        B. vietnamensis        B. vireti        B. vulcani        B. wakoensis        B. weihenstephanensis        B. xiamenensis        B. xiaoxiensis        B. zhanjiangensis          

     The above description is for the purpose of illustrating and not limiting the present invention, and teaching the person of ordinary skill in the art how to practice the invention. It is not intended to detail all those obvious modifications and variations of it which will become apparent to the skilled worker upon reading the description. It is intended, however, that all such obvious modifications and variations be included within the scope of the present invention as defined in the appended claims. The claims are meant to cover the claimed components and steps in any sequence which is effective to meet the objectives there intended, unless the context specifically indicates the contrary. 
     The patents, papers, and book excerpts cited above are hereby incorporated herein by reference in in their entireties.