MXPA05002873A

MXPA05002873A - Ammonia-oxidizing bacteria and methods of using and detecting the same.

Info

Publication number: MXPA05002873A
Application number: MXPA05002873A
Authority: MX
Inventors: Timothy A Hovanec
Original assignee: Aquaria Inc
Priority date: 2002-09-19
Filing date: 2003-09-10
Publication date: 2005-06-22
Also published as: AU2003272297A1; JP4470025B2; BR0314125A; WO2004026772A3; AU2003272297B2; WO2004026772A2; JP2010057494A; JP2006500027A; EP1551773A2; CA2499482A1

Abstract

Described herein are novel ammonia-oxidizing bacteria as well as isolated nucleotide sequences representative of 16S rDNA of these ammonia-oxidizing bacteria. Particular bacteria of the present invention are tolerant of freshwater environments, saltwater environments or both. Furthermore, in various embodiments, various bacteria of the present invention are capable of surviving a freeze-drying process, and may remain viable thereafter. Compositions including various combinations of the bacteria are further described, as are polymerase chain reaction (PCR) primers and oligonucleotide probes that may be used to detect these bacteria based on their 16S rDNA. Methods for preventing or alleviating the accumulation of ammonia in aqueous environments, such as aquaria and wastewater are also provided, using the ammonia-oxidizing bacteria of the present invention. Methods for detecting the bacteria of the present invention are also provided. Such methods may be effected by any conventional methology, such as with a DNA chip.

Description

OXIDIZING BACTERIA OF AMMONIA AND METHODS TO USE AND DETECT THE SAME FIELD OF THE INVENTION The invention relates generally to ammonia oxidants and specifically to bacteria capable of oxidizing ammonia to nitrite.

BACKGROUND OF THE INVENTION Ammonia is the main nitrogenous waste product of teleosts and many invertebrates in seawater and freshwater. Ammonia results from the removal or transamination of the proteins that the organism receives through its diet. However, high concentrations of ammonia can be toxic to many of these same aquatic organisms. In natural systems, such as lakes, rivers and oceans, the concentration of ammonia rarely reaches per udicial levels because the density of fish (and other organisms) per body of water is low. However, in man-made aquatic systems such as aquaculture tanks for breeding, tanks, canals and nurseries plus aquariums, public and private, ammonia can reach toxic concentrations, often very quickly. One reason for this is that in the systems mentioned above the density of the fish REF. : 162508 can be very large in relation to the small amount of water. Another reason is that in many of these systems the water does not change continuously; instead it is recirculated through the system with only periodic partial changes of water. Therefore, most aquaculture and aquarium systems use filtration, in one form or another, to maintain a degree of water quality that is appropriate for the maintenance and growth of aquatic organisms. A major component of such a filtration unit is the biological filter. The biological filter gets its name from the fact that it acts as a substrate or site for the growth of bacteria that have the ability to convert, through oxidation, ammonia to another compound-nitrite. The high concentrations of nitrite can also be toxic, but there are other species of bacteria that grow in the biological filter and oxidize the nitrite to nitrate, such as those described in U.S. Pat. num. 6,268,154, 6,265,206 and 6,207,440, each of which is incorporated by reference in its entirety as if it were fully established. Nitrate is not considered toxic to aquatic organisms except in extreme cases of very high concentrations. There are other situations or applications that use biological filters. These include sewage treatment facilities, wastewater treatment facilities and drinking water filtration plants. While each will have its own particular reason for using a biological filter, the goal is the same: the conversion of toxic inorganic nitrogen compounds to less harmful substances of inorganic nitrogen. It is necessary for many facilities with biological filtration that meet the National Recommended Water Quality Criteria as established by the Environmental Protection Agency (EPA) of the United States of America. Oxidation of ammonia to nitrite is a bacterially mediated process. Specifically, it is a two-step oxidation process that involves the conversion of ammonia to nitrite according to the following equations: NH3 + 02 + H20 + 2e ~? NH2OH + H20 (1) NH2OH + H20 - N02"+ 5H + + 4e ~ (2) The most commonly studied ammonia oxidation bacteria (AOB) is Nitrosomonas europaea. It was originally isolated from soils and was assumed to be the active AOB in the aquaculture facilities (Wheaton, FW 1977. Aquacultural Engineering, John Wiley &Sons, Inc. New York), in the wastewater treatment facilities (Painter, HA 1986. Nitrification in the treatment of sewage and waste-waters, in Nitrification JI Prosser ed. IRL Press, Oxford) and in aquariums (Spotte, S. 1979. Seawater Aquariums - The Captive Environment. Wiley-Interscience. New York). These references and all other references cited herein are incorporated by reference in their entirety as set forth in full. However, recent research carried out with modern molecular methods that use the uniqueness of the DNA sequence of an organism (or group of organisms) has shown that N. europaea and its close relatives were the lower detection limits in the environments of freshwater aquariums (Hovanec, TA and EF DeLong, 1996. Comparative analysis of nitrifying bacteria associated with freshwater and marine aquaria, Appl. Environ Microbiol. 62: 2888-2896). Other research has shown that N. europaea is not the dominant AOB in wastewater treatment facilities (Juretschko, S. et al 1998. Combined molecular and conventional analyzes of nitrifying bacterium diversity in activated sludge: Nitrosococcus mobilis and Ni rospira -like bacteria as dominant populations, Appl. Environ Microbiol. 64: 3042-3051). In addition, an environmental factor of particular importance with aquarium environments and wastewater treatment is sanitation, and, more specifically, the numerous physicochemical differences between freshwater and saltwater environments. The distinction between the different AOBs with respect to their capacity to tolerate such dramatic changes in the local environment is critical in the design of these systems and the implementation of the AOBs in them. Also, a tolerance demonstrated by a particular AOB to a saltwater environment can make the AOB appropriate for use in the aquarium and wastewater environments, and, in addition, a tolerance to withstand the change in the environment of the aquarium. Fresh water and salt water can have even more considerable implications. In addition, the storage and transport of AOB is often limited to liquid media and the like, potentially inconvenient, due to, at least in part, the inability of different strains of AOB to resist a freeze drying process. This process makes it possible to formulate a volume of AOB in a solid powder, freeze-dried or similar composition that can be tolerant to greater fluctuations in, for example, temperature, and can be correspondingly more practical for the purposes of transportation and handling in a marketed product, or similar considerations and to maintain a prolonged shelf life. Thus, there is a need in the art for the identification of AOBs, particularly those that are capable of tolerating a salt water environment and / or salt water and fresh water environments. There is also a need in the art for AOBs that remain viable after being subjected to a freeze drying process.

BRIEF DESCRIPTION OF THE INVENTION In one embodiment of the present invention, isolated bacteria or strains of bacteria capable of oxidizing ammonia to nitrite are provided. In one embodiment, the 16S rDNA of the bacteria or similar strains has the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEC ID NO: 19 or SEQ ID NO: 20. The nucleotide sequences described as SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 are examples of AOB similar to Nitrosomonas aestuarii. In different embodiments, the 16S rDNA of bacteria or bacterial strains has the nucleotide sequence of SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 (ie, AOB similar to Nitrosomonas aestuarii), SEC ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or a variant thereof which is at least 96% similar, at least 97% similar, at least 98% similar or at least 99% similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: twenty.

The present invention also includes nucleic acid sequences and bacteria with sequences having the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 or a variant thereof, which is at least 96% similar, at least 97% similar, at least 98% similar or at least 99% similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20. For the purposes of this application, "96% similar" means that substitutions of the simple base can occur in up to 4% of the bases, "97% similar" means that substitutions of the simple base can occur in up to 3% of the bases, "98% "similar" means that substitutions of the simple base can occur in up to 2% of the bases and "99% similar" means that substitutions of the simple base can occur in up to 1% of the bases. The present invention also includes methods for decreasing ammonia accumulation in a medium. The methods include a step of placing in the medium a sufficient amount of a bacterial strain or a composition comprising a bacterial strain, wherein the 16S rDNA of the bacterial strain has the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO. : 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 or a variant thereof, which is at least 96% similar, by at least 97% similar, at least 98% similar or at least 99% similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18 , SEQ ID NO: 19 or SEQ ID NO: 20. The present invention also includes a method for detecting and determining the amount of bacteria in a medium capable of oxidizing ammonia to nitrite. The method includes providing a detectably labeled probe of the present invention, isolating the total DNA that forms the medium, exposing the isolated DNA to the probe under conditions wherein the probe hybridizes only to the nucleic acid of the bacteria when the rDNA 16 the bacteria has a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 and detect and measure the probe to detect and measure the amount of bacteria.

BRIEF DESCRIPTION OF THE FIGURES Figure 1 illustrates the phylogenetic relationships of three bacterial strains (ie, those represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3) and a sub-layer (i.e. strain represented by SEQ ID NO: 4) inferred from the comparative analysis of the 16S rDNA sequences according to one embodiment of the present invention. The tree is based on the distance analysis of the neighboring union of substances that contain a minimum of 1430 nucleotides. Figure 2 illustrates a gel electrophoresis of the denaturing gradient (DGGE) of selected crop biomass and ammonia oxidizing bacteria represented by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, according to one embodiment of the present invention. Figure 3 illustrates a DGGE demonstrating the uniqueness of the bacterial strains represented by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, according to one embodiment of the present invention. invention. There are two replicates of each type of bacteria mentioned above along with the extracts of three pure cultures of bacteria that oxidize ammonia. Figures 4A-4D 'illustrate the average ammonia and nitrite trends for the Bacterial Additives test VI for the bacterial strains represented by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, according to one embodiment of the present invention. Figures 5A-5D 'illustrate the mean ammonia and nitrite trends for the Bacterial Additives test VII for the bacterial strains represented by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, according to one embodiment of the present invention.

Figure 6 illustrates the phylogenetic relationships of two bacterial strains (ie, those represented by SEQ ID NO: 18 and SEQ ID NO: 19) and one sub-canopy (ie, that represented by SEQ ID NO: 20) inferred of the comparative analyzes of the 16S rDNA sequences according to one embodiment of the present invention. The tree further represents the relationship between the two bacterial strains represented by SEQ ID NO: 18 and SEQ ID NO: 19 and the bacterial strains represented by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. The tree is based on the distance analysis of the neighboring union of substances containing a minimum of 1430 nucleotides. Figure 7 illustrates a gel electrophoresis of the denaturing gradient (DGGE) of the biomass of freshwater cultures selected from ammonia oxidizing bacteria represented by SEQ ID NO: 1, SEQ ID NO: 3 and SEQ ID NO. : 4 together with the seawater cultures of the bacteria that oxidize the ammonia represented by SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 and the pure cultures of the bacteria that oxidize ammonia Nitrosomonas europaea, Nitrosomonas multiformis and Nitrosomonas cryotolerans. Figure 8 illustrates the trends of the average concentration of ammonia for the Bacterial Additive VIII test for freshwater bacterial strains represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 and the bacterial strains of seawater represented by SEQ ID NO: 18, SEQ ID NO: 19 and two strains similar to N. halophila according to one embodiment of the present invention, together with two commercially available mixtures of nitrifying bacteria. Figure 9 illustrates the trends of the average concentration of ammonia for aquariums in the Bacterial Additives IX test that were dosed with the bacterial strains of seawater represented by SEQ ID NO: 18, SEQ ID NO: 19 and two similar strains N. halophila according to one embodiment of the present invention. Figure 10 illustrates the trends of the average concentration of ammonia for the Bacterial Additives X test. Two bacterial mixtures of bacterial strains of seawater represented by SEQ ID NO: 18, SEQ ID NO: 19 and two N-like strains. halophila were tested against aquariums not inoculated according to one embodiment of the present invention.

DETAILED DESCRIPTION OF THE INVENTION The present invention is based on the discovery of new bacterial strains that are capable of oxidation of ammonia in freshwater and / or saltwater environments, and that can also survive and remain viable after a drying process. by frozen. The embodiments of the present invention describe methods for using bacterial strains. The present invention provides an isolated bacterial strain or a biologically pure culture of a bacterial strain capable of oxidizing ammonia to nitrite, wherein the 16S rDNA of the bacterial strain includes the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 22 as shown in Tables 1 to 7.

Table 1: Sequence for the bacterium that oxidizes Ammonia Type A of AOB. Represented by R7clonl40. SEC ID NO: 1.

ATTGAACGCTGGCGGCATGCTTTACACATGCAAGTCGAACGGCAGCACGGAT GCTTGCATCTGGTGGCGAGTGGCGGACGGGTGAGTAATGCATCGGAACGTAT CCAGAAGAGGGGGGTAACGCATCGAAAGATGTGCTAATACCGCATATACTC TAAGGAGGAAAGCAGGGGATCGAAAGACCTTGCGCTTTTGGAGCGGCCGATG TCTGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCAGTAGT TGGTCTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCC TACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGCGCAAGCCTGATCCAG CAATGCCGCGTGAGTGAAGAAGGCCTTCGGGTTGTAAAGCTCTTTCAGTCGA GAAGAAAAGGTTACGGTAAATAATCGTGACTCATGACGGTATCGACAGAAG AAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGC GTTAATCGGAATTACTGGGCGTAAAGGGTGCGCAGGCGGCTTTGTAAGTCAG ATGTGAAATCCCCGGGCTTAACCTGGGAATTGCGTTTGAAACTACAAGGCTA GAGTGTGGCAGAGGGAGGTGGAATTCCATGTGTAGCAGTGAAATGCGTAGAG ATATGGAAGAACATCGATGGCGAAGGCAGCCTCCTGGGTTAACACTGACGCT CATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACG CCCTAAACGATGTCAACTAGTTGTTGGGCCTTATTAGGCTTGGTAACGAAGC TAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAA AGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCA CACGTAATACAATGGCGCGTACAGAGGGTTGCCAACCCGCGAGGGGGAGCTA ATCTCAGAAAGCGCGTCGTAGTCCGGATCGGAGTCTGCAACTCGACTCCGTG AAGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCC CGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTCACCAGAAG CAGGTAGTCTAACCGTAAGGAGGGCGCTTGCCACGGTGAGATTCATGACTGG GGTG Table 2: Sequence for the bacteria that oxidizes Type A ammonia of AOB. Represented by R7clonl87. SEC ID NO: 2 ATTGAACGCTGGCGGCATGCTTTACACATGCAAGTCGAACGGCAGCACGGAT GCTTGCATCTGGTGGCGAGTGGCGGACGGGTGAGTAATGCATCGGAACGTAT CCAGAAGAGGGGGGTAACGCATCGAAAGATGTGCTAATACCGCATATACTC TAAGGAGGAAAGCAGGGGATCGAAAGACCTTGCGCTTTTGGAGCGGCCGATG TCTGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCAGTAGT TGGTCTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCC TACGGGAGGCAGCAGTGGGGAATTITGGACAATGGGCGCAAGCCTGATCCAG CAATGCCGCGTGAGTGAAGAAGGCCTTCGGGTTGTAAAGCTCTTTCAGTCGA GAAGAAAAGGTTACGGTAAATAATCGTGACCCATGACGGTATCGACAGAAG AAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGC GTTAATCGGAATTACTGGGCGTAAAGGGTGCGCAGGCGGCCTTGTAAGTCAG ATGTGAAATCCCCGGGCTTAACCTGGGAATTGCGTTTGAAACTACAAAGCTA GAGTGTGGCAGAGGGAGGTGGAATTCCATGTGTAGCAGTGAAATGCGTAGAG ATATGGAAGAACATCGATGGCGAAGGCAGCCTCCTGGGTTAACACTGACGCT CATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACG CCCTAAACGATGTCAACTAGTTGTTGGGCCTTATTAGGCTTGGTAACGAAGC TAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAA AGGAAITGACGGGGACCCGCACAAGCGGTGGATTATGTGGATTAATTCGATG CAACGCGAAAAACCTIACCTACCCTTGACATGTAGCGAATTTTCTAGA GAT AGATTAGTGCTTCGGGAACGCTAACACAGGTGCTGCATGGCTGTCGTCAGCT CGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATT AATTGCCATCATTTGGTTGGGCACTTTAATGAGACTGCCGGTGACAAACCGG AGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGOOTAGOGCTTCA CACGTAATACAATGGCGCGTACAGAGGGTTOCCAACCCGCOAOGGGGAGCTA ATCTCAGAAAGCOCGTCGTAGTCCGGATCGGAGTCTGCAACTCGACTCCGTG AAGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTOAATACGTTCC CGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTCACCAGAAG CAGGTAGTCTAACCGTAAGGAGGGCGCTTGCCACGGTGAGATTCATGACTGG GGTG Table 3: Sequence for the bacterium that oxidizes the Type B ammonia of AOB. Represented by R3clon5. SEC ID NO: 3.

ATTGAACGCTGGCGGCATGCTTTACACATGCAAGTCGAACGGCAGCACGGGG GCAACCCTGGTGGCGAGTGGCGAACGGGTGAGTAATACATCGGAACGTATCT TCGAGGGGGGGATAACGCACCGAAAGGTGTGCTAATACCGCATAATCTCCAC GGAGAAAAGCAGGGGATCGCAAGACCTTGCGCTCTTGGAGCGGCCGATGTCT GATIAGCTAGTIGGTGAGGTAATGGCTTACCAAGGCGACGATCAGTAGCTGG TCTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTAC GGGAGGCAGCAGTGGGGAATTTTGGACAATGGGGGAAACCCTGATCCAGCCA TGCCGCGTGAGTGAAGAAGGCCTTCGGGTTGTAAAGCTCTTTCAGCCGGAAC GAAACGGTCACGGCTAATACCCGTGACTACTGACGGTACCGGAAGAAGAAG CACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTT AATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTTTGTAAGTCAGATG TGAAAGCCCCGGGCTTAACCTGGGAACTGCGTTTGAAACTACAAGGCTAGAG TGTGGCAGAGGGGGGTGGAATTCCACGTGTAGCAGTGAAATGCGTAGAGATG TGGAGGAACACCGATGGCGAAGGCAGCCCCCTGGGTTAACACCGACGCTCAG GCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCC TAAACGATGTCAACTAGTTGTCGGGTCTTAACGGACTTGGTAACGCAGCTAA CGCGTGAAGTTGGCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGG AATTGACGGGGACCCGCACAAGCGGTGGATTATGTGGATTAATTCGATGCAA CGCG-CCTTACCTACCCTTGACATGTACCGAAGCCCGCCGAGAGGTG G GTGTGCCCGAAAGGGAGCGGTAACACAGGTGCTGCATGGCTGTCGTCAGCTC GTGTCGTGAGATGTTGGOTTAAGTCCCGCAACGAGCGCAACCCTTGTCATTA ATTGCCATCATTCAGTTGGGCACTTTAATGAAACTGCCGGTGACAAACCGGA GGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCAC ACGTAATACAATGGCGCGTACAGAGGGTTGCCAACCCGCGAGGGGGAGCTAA TCTCAGAAAGCGCGTCGTAGTCCGGATCGGAGTCTGCAACTCGACTCCGTGA AGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCCC GGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTCACCAGAAGC AGGTAGTCTAACCGCAAGGAGGGCGCTTGCCACGGTGAGATTCATGACTGGG GTG Table 4: Sequence for the bacterium that oxidizes the Type A ammonia of AOB. Represented by R5clon47. SEC ID NO: 4 ATTGAACGCTGGCGGCATGCTTTACACATGCAAGTCGAACGGCAGCGGGGGC TTCGGCCTGCCGGCGAGTGGCGAACGGGTGAGTAATACATCGGAACGTGTCC TTAAGTGGGGAATAACGCATCGAAAGATGTGCTAATACCGCATATCTCTGA GGAGAAAAGCAGGGGATCGCAAGACCTTGCGCTAAAGGAGCGGCCGATGTCT GATTAGCTAGTTGGTGGGGTAAAGGCTTACCAAGGCAACGATCAGTAGTTGG TCTGAGAGGACGACCAACCACACTGGGACTGAGACACGGCCCAGACTCCTAC GGGAGGCAGCAGTGGGGAATTTTGGACAATGGGCGAAAGCCTGATCCAGCCA TGCCGCGTGAGTGAAGAAGGCCTTCGGGTTGTAGAGCTCTTITAGTCAGAAA GAAAGAATCATGATGAATAATTATGATITATGACGGTACTGACAGAAAAAG CACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTT AATCGGAATIACTGGGCGTAAAGGGTGCGCAGGCGGTTTTGTAAGTCAGATG TGAAAGCCCCGGGCTTAACCTGGGAATTGCGTTTGAAACTACAAGGCTAGAG TGCAGCAGAGGGGAGTGGAATTCCATGTGTAGCAGTGAAATGCGTAGAGATG TGGAAGAACACCGATGGCGAAGGCAGCTCCCTGGGTTGACACTGACGCTCAT GCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCC TAAACGATGTCAACTGGTTGTCGGATCTAATTAAGGATTTGGTAACGTAGCT AACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAA AGGAATTGACGGGGACCCGCACAAGCGGTGGATIATGTGGATTAATTCGATG CAACGCGAAAAACCTTACCTACCCTTGACATGCTTGGAATCTAGTGGAGAC ATAAGAGTGCCCGAAAGGGAGCCAAGACACAGGTGCTGCATGGCTGTCGTCA GCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTC ACTAATTGCTATCATTCTAAATGAGCACTTTAGTGAGACTGCCGGTGACAA ACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGG CTTCACACGTAATACAATGGCGTGTACAGAGGGTTGCCAACCCGCGAGGGGG AGCCAATCTCAGAAAGCACGTCGTAGTCCGGATCGGAGTCTGCAACTCGACT CCGTGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATAC GTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGTTTTCACC AGAAGCAGGTAGTTTAACCGTAAGGAGGACGCTTGCCACGGTGGGGGTCATG ACTGGGGTG Table 5: Sequence for AOB similar to Nitrosomes aestuarll represented by P4clon42. SEC ID NO: 18.

TTGATCATGGCTCAGATTGAACGCTGGCGGCATGCTTTACACATGCAAGTCGAACGG CAGCACGGGTGCTTGCACCTGGTGGCGAGTGGCGGACGGGTGAGTAATGCATCGGAA CGTGTCCAGAAGTGGGGGATAACGCATCGAAAGATGTGCTAATACCGCATATTCTCT ACGGAGGAAAGCAGGGGATCGAAAGACCTTGTGCTTTTGGAGCGGCCGATGCCTGAT TAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCAACGATCAGTAGTTGGTCTGAGAG GACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAG TGGGGAATTTTGGACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGAGTGAAGA AGGCTTCGGGTTGTAAAGCTCTTTCAGTCGAGAAGAAAAGGTTGTGACTAATAATCA CAACTTATGATGGTACCGACAGAAGAAGCACCGGCTAACTACGTGCCAGCAGCCGCG GTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGGGTGCGCAGGC GGCTTTGTAAGTCAGATGTGAAATCCCCGGGCTTAACCTGGGAATTGCGTTTGAAACT ACAAAGCTAGAGTGTAGCAGAGGGGGGTGGAATTCCATGTGTAGCAGTGAAATGCG TAGAGATATGGAAGAACATCGATGGCGAAGGCAGCCCCCTGGGTTAACACTGACGCT CATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTA AACGATGTCAACTAGTTGTTGGGCCTTACTAGGCTTGGTAACGTAGCTAACGCGTGA AGTTGACCGCCTGGGGAGTACGGTCGCAGGATTAAAACTCAAAGGAATTGACGGGG ACCCGCACAAGCGGTGGATIATGTGGATTAATTCGATGCAACGCGAAAAACCTTACC TACCCTTGACATGTAG CGAATATTTTAGAGATAAAATAGTGCCTTCGGGAACGCTAA CACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGC AACGAGCGCAACCCTIGTCATTAATTGCCATCATTTAGTTGGGCACTITAATGAGACT GCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGG GTAGGGCTTCACACGTAATACAATGGCGCGTACAGAGGGTTGCCAACCCGCGAGGGG GAGCTAATCTCAGAAAGCGCGTCGTAGTCCGGATCGGAGTCTGCAACTCGACTCCGT GAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCCCGGG TCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTCACCAGAAGCAGATAGTC TAACCGTAAGAGGGCGTTTGCCACGGCGAGATTCATGACTGG Table 6: Sequence for AOB s milar to Nitrosomes estuaril represented by P4clon31. SEQ ID NO: 19 AGTTTGATCATGGCTCAGATTGAACGCTGGCGGCATGCTTTACACATGCAAGTCGAA CGGCAGCACGGGTGCTTGCACCTGGTGGCGAGTGGCGGACGGGTGAGTAATGCATCG GAACGTGTCCGGAAGTGGGGGATAACGCATCGAAAGATGTGCTAATACCGCATATTC TCTACGGAGGAAAGCAGGGGATCGAAAGACCTTGTGCTTTTGGAGCGGCCGATGCCT GATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCAACGATCAGTAGTTGGTCTGA GAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAG CAGTGGGGAATTTTGGACAACGGGCGAAAGCCTGATCCAGCAATGCCGCGTGAGTGA AGAAGGCCTTCGGGTTGTAAAGCTCTTTCAGTCGAGAAGAAAAGGTTGTGACTAATA ATCACAACTTATGACGGTACCGACAGAAGAAGCACCGGCTAACTACGTGCCAGCAGC CGCGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGGGTGCGC AGGCGGCTTTGTAAGTCAGATGTGAAATCCCCGGGCTTAACCTGGGAATTGCGTTTG AAACTACAAAGCTAGAGTGTAGCAGAGGGGGGTGGAATTCCATGTGTAGCAGTGAA ATGCGTAGAGATATGGAAGAACATCGATGGCGAAGGCAGCCCCCTGGGTTAACACTG ACGCTCATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACG CCCTAAACGATGTCAACTAGTTGTTGGGCCTTACTAGGCTTGGTAACGTAGCTAACGC GTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACG GGGACCCGCACAAGCGGTGGATTATGTGGATTAATTCGATGCAACGCGAAAAACCTT ACCTACCCTTGACAT GTAGCGAATATTTTAGAGATAAAATAGTGCCTTCGGGAACGC TAACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCC CGCAACGAGCGCAACCCTTGTCATTAATTGCCATCATTTAGTTGGGCACTTTAATGAG ACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTA TGGGTAGGGCTTCACACGTAATACAATGGCGCGTACAGAGGGTTGCCAACCCGCGAG GGGGAGCTAATCTCAGAAAGCGCGTCGTAGTCCGGATCGGAGTTAGCAACTCGACTC CGTGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCCC GGGCCTTGTACACACCGCCCGTCACACCATGGAAGTTGGCTGCACCAGAAGTAGGTT GTCTAACCCTCGGGAGGACGCTTACCACGGTGTGGTCAATGACTTGGGGTGAAGTCG TAACAAGGTAA Table 7: Sequence for AOB simi dollars to Nitrosornas aescuarii. represented by BF16clon57. SEC ID NO: 20 GTTTGATCATGGCTCAGATTGAACGCTGGCGGCATGCTTTACACATGCAAGTCGAAC GGCAGCACGGGTGCTTGCACCTGGTGGCGAGTGGCGGACGGGTGAGTAATGCATCGG AACGTGTCCAGAAGTGGGGGATAACGCATCGAAAGATGTGCTAATACCGCATATTCT CTACGGAGGAAAGCAGGGGATCGAAAGACCTTGTGCTTTTGGAGCGGCCGATGCCTG ATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCAACGATCAGTAGTTGGTCTGAG AGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGC AGTGGGGAATTTTGGACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGAGTGAA GAAGGCCTTCGGGTTGTAAAGCTCTTTCAGTCGAGAAGAAAAGGTTGTGACTAATAA TCACAACTTATGACGGTACCGACAGAAGAAGCACCGGCTAACTACGTGCCAGCAGCC GCGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGGGTGCGCA GGCGGCTTTGTAAGTCAGATGTGAAATCCCCGGGCTTAACCTGGGAATTGCGTTTGA AACTACAAAGCTAGAGTGTAGCAGAGGGGGGTGGAATTCCATGTGTAGCAGTGAAA TGCGTAGAGATATGGAAGAACATCGATGGCGAAGGCAGCCCCCTGGGTTAACACTGA CGCTCATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGC CCTAAACGATGTCAACTAGTTGTTGGGCCTTACTAGGCTTGGTAACGTAGCTAACGCG TGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGG GGACCCGCACAAGCGGTGGATTATGTGGATTAATTCGATGCAACGCGAAAAACCTTA CCTACCCTTGACATGTAGCGAATATTTTAGAGATAAAATAGTGCCTTCGGGAACGCT AACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCC GCAACGAGCGCAACCCTTGTCATTAATTGCCATCATTTAGTTGGGCACTTTAATGAGA CTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTAT GGGTAGGGCTTCACACGTAATACAATGGCGCGTACAGAGGGTTGCCAACCCGCGAGG GGGAGCTAATCTCAGAAAGCGCGTCGTAGTCCGGATCGGAGTCTGCAACTCGACTCC GTGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCCCG GGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTCACCAGAAGCAGATAG TCTAACCGTAAGGAGGGCGTTTGCCACGGTGAGATTCATGACTGGGGTGAAGTCGTA ACAATTTA For the purposes of the present invention, an isolated bacterial strain is one that has undergone some degree of purification from its natural environment. A culture of a bacterium is considered to be biologically pure if at least 20% of the bacteria are from a bacterial strain. However, it is preferable if the culture is at least 33% pure, more preferable if the culture is at least 45% pure and more preferable if the culture is at least 90% pure. The bacterial strains of the present invention can also be combined with each other, other species of bacteria, nutrients and / or other components to provide a composition for the purpose of maintaining or purifying the water-containing media. It may be desirable, for example, to combine the bacteria of the present invention with bacteria capable of removing other contaminants or undesirable compounds from the media containing water. Examples of these bacteria include bacteria that oxidize nitrite (chemolithoautotrophic bacteria that oxidize nitrite to nitrate), heterotrophic bacteria (which mineralize organic material into ammonia and other substances) and other bacteria that are well known to those skilled in the art. The bacteria that oxidize nitrite are known from the phylum of Nitrospira bacteria, and the alpha, gamma and delta subdivisions of the Proteobacteria. Examples include the species of the genera Nitrospira, Nitrospina and Nitrobacter. The bacteria that reduce nitrate are well known from the genera Azoarcus, Pseudomonas and Alcaligenes. The heterotrophic bacteria are known from the genera Bacillus, Planctomyces, Pseudomonas and Alcaligenes. These are available from known sources (eg, American Type Culture Collection, 10801 University Blvd., Manassas VA 20100, USA) or can be isolated directly from aquarium biofilters. For example, the bacterial strains of the present invention can be combined with the bacteria that oxidize the nitrite, so that the ammonia present in the aqueous system would be oxidized to nitrite and the nitrite would be oxidized to nitrate. Another example would be to combine the bacterial strain of the present invention with aerobic or anaerobic denitrification bacteria. In this case, the nitrate that is produced by the interaction of the bacterial strains of the present invention with the bacteria that oxidize the nitrite would be reduced to dinitrogen or other nitrogen-based products. A third example would be to combine the bacterial strain of the present invention with heterotrophic bacteria that mineralize the organic matter into simpler inorganic substances which, subsequently, can be used as substrates by the bacterial strains of the present invention. The present invention also provides a mixture comprising a concentrated bacterial strain capable of oxidizing ammonia to nitrite, wherein the 16S rDNA of the bacteria has a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO. : 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 or a variant thereof, which is at least 96% similar, at least 97% similar, at least 98% similar or at least 99% similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20. In accordance with this embodiment of the invention, the bacterial strain is considered to be concentrated if the bacterial strain is present in a concentration that is greater than its concentration presented in nature. In general, the concentration of the bacterial strain will be at least 20% of the total cells in the sample, determined by standard techniques, such as molecular probing using fluorescent in situ hybridization (FISH) techniques. , which will be known to those skilled in the art, using the controls and the appropriate enumeration methods. More specifically, the concentration of the bacterial strain would be 33% or more of the total cells, still more preferably 45% and more preferably 90% or more of the total cells. However, it may be preferable that more than one of the bacteria have a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEC ID NO: 19 or SEQ ID NO: 20 in the mixture. In this case, the percentages established above refer to the percentage of the total AOBs in the mixture, understanding that the balance of the cell population may be comprised of bacteria that oxidize nitrite or other types of bacteria. In particular, while not wishing to be related to the theory, of the different bacterial strains described according to the present invention, the strains represented by SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 are believed to be tolerant especially to saltwater environments; although these strains can also be used in freshwater environments, and are believed to work effectively in it. Bacterial strains and mixtures that incorporate the strains in addition to the strains represented by SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, can also tolerate salt water environments to an appreciable extent, yet in a preferred embodiment of the present invention, are strains represented by SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 which are included in a salt water environment to oxidize ammonia to nitrite. In addition, while any of the bacterial strains of the present invention can be frozen-dried, the strains represented by SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 are believed to be particularly process tolerant. of drying by freezing, as evidenced by its ability to remain viable after such process and to oxidize the ammonia to nitrite after this process. Thus, in a preferred embodiment of the present invention, the bacterial strains represented by SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 can be dried by freezing and subsequently used to oxidize the ammonia to nitrite either in freshwater or saltwater environments. It is understood that the bacterial strains and mixtures of the present invention may be in the form of powder, liquid, a frozen form, a freeze dried form or any other suitable form, which can be easily recognized by one skilled in the art. These are commonly referred to as "commercial additives", and may include, but are not limited in any way to: (1) a liquid form, wherein one or more strains are in a liquid solution containing inorganic salts or organic compounds, so that the viability of the cells is not destroyed during the course of storage; (2) a frozen form, wherein one or more of the strains are in a liquid mixture as above, optionally including cryoprotectant compounds to prevent cell lysis, which is frozen and stored at a temperature at or below 0 ° C ( 32 ° F); and (3) a powder form, which has been produced by freeze drying or other means, wherein the dehydrated form of one or more of the strains or mixture can be stored at normal room temperature without loss of viability. The obtaining of an appropriate form of the bacterial strain and the mixture of the present invention are within those experienced in the art from the point of view of the above description. It is also understood that the bacterial strains and the mixture of the present invention can be used alone or in combination with other components. Examples of the components include, but are not limited to, bacteria that oxidize nitrite, heterotrophic bacteria that oxidize nitrite, heterotrophic bacteria that oxidize ammonia and the like. All forms of the biologically pure bacterial strain can also contain nutrients, amino acids, vitamins and other components that serve to preserve and promote the growth of the bacterial strain. The bacterial strains and mixtures and compositions of the present invention can be used in freshwater aquariums, seawater aquariums and wastewater to reduce ammonia accumulation. They can be used in a bio-remediation process to reduce the level of contamination caused by ammonia. A bio-remediation process, also called bio-augmentation, includes, but is not limited to, the supplemental addition of microorganisms to a system (for example, a site where biological or chemical contamination has occurred) for the purposes of promoting or stabilize the biological and / or chemical processes that result in the change of one or more forms of chemical compounds present in the original system. Therefore, one aspect of the present invention provides a method for reducing ammonia accumulation in a medium. The method includes a step of placing in the medium a sufficient amount of a bacterial strain capable of oxidizing ammonia to nitrite to reduce ammonia accumulation in the medium, wherein the 16S rDNA of the bacterial strain has a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 or a variant thereof, which is at least 96% similar, at least 97% similar, at least 98% similar or at least 99% similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20. The amount of the bacterial strain (s) is sufficient if the added bacteria can reduce or prevent the accumulation of ammonia in the medium. In general, the addition of one or more of the bacterial strains of the invention to a freshwater or saltwater aquarium is expected to reduce the maximum concentration of ammonia by at least 50% of the level that would be expected in the absence of the (s) bacterial strain (s). In another embodiment of the invention, a method for reducing ammonia accumulation in a medium includes placing in the medium a sufficient amount of a composition, as described herein, to maintain or purify the water-containing media. The composition may comprise one or more bacterial strains capable of oxidizing ammonia to nitrite, wherein the 16S rDNA of the bacterial strain or strains has a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 or a variant thereof, which is at least 96% similar, at least 97% similar, by at least 98% similar or at least 99% similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEC ID NO: 20. It will be appreciated that the actual levels achieved in a given facility will be a function of the size and contents of the systems (ie the number of fish, plants, etc.). In a newly established 37-liter aquarium with ten fish, the ammonia concentration can reach 7 mg / L or more without the addition of the bacterial strain, while the maximum level can be reduced to approximately 2 mg / L by adding the bacterial strain. In general, the maximum concentration of ammonia would not be expected to exceed 3 mg / L if the bacterial strain of the invention were added to such a system. When the system reaches a steady state, ammonia levels fall below 0.5 mg / L, a process that occurs more rapidly when the bacterial strain of the invention is present. In one embodiment of the present invention, the bacterial strains of the present invention are placed directly in a medium, such as, but not limited to, freshwater aquariums, seawater aquariums and wastewater. In another embodiment of the present invention, the bacteria can grow in a rotary biological contactor and then be placed in the medium. In a different embodiment, the bacteria of the present invention can be placed in a biofilter unit contained in the medium. In another embodiment, the bacteria of the present invention can be immobilized in an immobilized polymer, such as, but not limited to, acrylamide, alginate or carrageenan. This bacterial polymeric material then attached can be placed in a filter or it can be placed in the filter stream of a suitable installation. As used herein, the term "aquarium" refers to a container that can be made of, in combination or in its entirety, but not limited to, glass, plastic or wood that houses water, and in which they are placed living aquatic organisms (such as fish, plants, bacteria and invertebrates) and the contents thereof. An aquarium can be for the purposes of exhibiting aquatic organisms, for short or long term accommodation, for scientific study, for transportation and other purposes. In general, a freshwater aquarium is an aquarium in which the liquid medium has a salinity of less than 15 parts per thousand. A saltwater aquarium in general is an aquarium in which the liquid medium has a salinity greater than 15 parts per thousand. The term "aquarium water" is used to refer to the medium that is contained within the aquarium and is associated with filtering systems, in which aquatic organisms reside. The aquarium water may contain a wide range of inorganic or organic chemicals and, therefore, may have a wide range of parameters, such as salt concentration, pH, total dissolved solids and temperature, to mention a few. As used herein, "wastewater" generally refers to a liquid medium that is the product of an industrial or human process. It may require treatment by one or more filtration methods to make it less harmful to the environment, so that it conforms to the discharge standards determined by the government agency. Wastewater can also be recycled so that it does not discharge into the environment. As used herein, a "biological filter" also called a "biofilter", usually refers to a type of filter whose purpose is to promote the growth of microorganisms or provide a substrate for the binding and growth of microorganisms. A biofilter can be part of an aquarium filtration system or a wastewater filtration system. As used herein, the term "rotary biological contactor" usually refers to a type of biofilter that rotates in water or medium. It can be completely or partially immersed in the water or medium. Those skilled in the art will recognize that rotary biological contactors as are modeled in U.S. Pat. 2,085,217; 2,172,067; 5,423,978; 5,419,831; 5,679,253; 5,779,885 and all continuations, improvements and foreign counterparts; each of which is incorporated herein by reference as if they were fully established. As used herein, "filtering silk" refers to a natural or synthetic multiple-strand material of irregular shape that can serve as a biofilter, a mechanical filter or a combination thereof. As used herein, "aquarium gravel" refers to a substrate commonly placed within, at the bottom of an aquarium. It may be composed of pieces of rock of irregular or regular shape, coral, plastic or other material. It can serve as a biofilter, a mechanical filter, for decorative purposes or a combination of these. As used herein, "filter sponge" refers to a natural or synthetic material that when used in an aquarium or as part of an aquarium filtration system, can serve as a mechanical filter, biofilter or both. As used herein, "plastic filter media" refers to a man-made material that serves as a biofilter, a mechanical filter, or both. It can be molded plastic or injection molded. In another embodiment, the nucleic acid sequences and the bacteria are also provided with the sequences having the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEC ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 or a variant thereof that is at least 96% similar, at least 97% similar, at least 98% similar or at least 99 % similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20. In another embodiment , nucleotide probes are provided to detect and measure the amount of bacteria of the present invention, which are present in a medium. The probes have the nucleotide sequences set forth in SEQ ID NO: 5, SEQ ID NO: 8 and SEQ ID NO: 21. The nucleotide probes of the present invention can be synthesized by methods that are known in the art. The nucleotide probes of the present invention can be labeled by any of the labels that can be detected. Examples of the appropriate labels include, but are not limited in any way to, radioactive labels, fluorescent labels and the like. Suitable labeling materials are commercially available and would be those known to those skilled in the art. Methods of labeling an oligonucleotide or a polynucleotide are also known to those skilled in the art (See, for example, Sambrook, J., EF Fritsch and T. Maniatis, Molecular Cloning-A Laboratory Manual, 2. sup.nd edition , 1989, Cold Spring Harbor Press). The nucleotide probes of the present invention are capable of hybridizing to 16S rDNA of the bacterial strain of the present invention. Therefore, the nucleotide probes of the present invention are very suitable for use in a method for detecting and determining the amount of bacteria of the present invention. In one aspect of the present invention, there is provided a method for detecting and determining the amount of bacteria capable of oxidizing ammonia to nitrite in a medium, wherein the 16S rDNA of the bacteria has a nucleotide sequence of SEQ ID NO: 1 , SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20. The method may include: (a) providing a labeled probe in a detectable manner of the present invention; (b) isolating total DNA from a medium; (c) exposing the isolated total DNA to the detectably labeled probe under conditions in which the probe hybridizes only to the nucleic acid of the bacteria, wherein the 16S rDNA of the bacteria has a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20; and (d) detecting and measuring the hybridized probe to detect and measure the amount of the bacteria. The probes of the present invention are represented by SEQ ID NO: 5, SEQ ID NO: 8 and SEQ ID NO: 21. A sequence that is at least 96% similar over the total length of any of the probes mentioned above. it can also be used to detect the bacteria of the present invention. These probes are further described in the following examples. The medium can be aquarium water, where the DNA is isolated from it. The medium may also contain a material such as aquarium gravel, sponge filter material, filter cloth or plastic filter media, but it is not considered to be limiting thereof. Therefore, DNA can be isolated from previous sources and others where these bacteria can be expected to be found. The method of the present invention can be performed in conjunction with a portion of DNA or similar tools known to those skilled in the art. A portion of DNA can include a solid support and a group of nucleotide derivatives or analogs attached to the solid support via a covalent bond. Detection of a nucleic acid fragment with a portion of DNA is generally performed using a probe oligonucleotide that is complementary to the nucleic acid fragment to be detected, by means of hybridization. The probe oligonucleotide is generally fixed on the solid support (e.g., solid substrate). In the detection processes, a nucleic acid fragment in a sample liquid can be provided with a fluorescent label or a radioisotope label and then the Sample liquid can be contacted with the probe oligonucleotide of the DNA portion. If the labeled nucleic acid fragment in the sample liquid is complementary to the probe oligonucleotide, the labeled nucleic acid fragment is combined with the probe oligonucleotide by hybridization. The labeled nucleic acid fragment attached to the DNA portion by hybridization with the probe oligonucleotide can then be detected by an appropriate detection method, such as by way of example, fluorometry or autoradiography, although other methods of detection can be used. The method may alternatively be performed in conjunction with a wide variety of automated processes, which will be readily recognized by those experienced in the art and implemented by routine experimentation. By way of example, the method of the present invention can be performed with DNA microarrays or proteins, biosensors, biosondes, capillary electrophoresis and real-time PCR to name a few common methodologies; although it will be easily appreciated by one experienced in the art that this list may include others. The detection method of the present invention provides an effective tool for monitoring and detecting the occurrence of bacteria capable of oxidizing ammonia to nitrite in a medium. The method also provides a tool for verifying commercial additives to determine the effectiveness of the additives, by measuring the occurrence or amount of the bacteria of the present invention. In another embodiment, the PCR primers that can be used to detect the bacteria and nucleotide sequences of the present invention. The pairs of PCR primers are represented by SEQ ID NO: 6 and SEQ ID NO: 7, SEQ ID NO: 9 and SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 and SEQ ID NO: 22 and SEQ ID NO: 23. A sequence that is at least 96% similar over the total length of any of the aforementioned PCR primers can also be used to detect the bacteria of the present invention. These PCR primers are further described in the following examples. It would be readily apparent to one of ordinary skill in the art that the variants of the oligonucleotide probes mentioned above and the PCR primers that can still be used to detect the bacteria and nucleic acid sequences of the present invention, are within the scope of the present invention. For example, a variant of any of the oligonucleotide or primer probes that differs from SEQ ID NO: 5, SEQ ID NO: 8, SEQ ID NO: 21, SEQ ID NO: 6, SEQ ID is embraced by the present invention. NO: 7, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 22 or SEQ ID NO: 23 due to one or more additions, deletions or substitutions nucleotides of the present invention. The present invention includes isolated bacteria isolated bacterial strains, bacterial cultures and nucleotide sequences comprising the sequences identified herein as SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEC ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, or variants of these sequences. Particularly, the preferred variants are those in which there is a high degree of similarity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20. The present invention includes variants that are at least 96% similar, at least 97% similar, at least 98% similar or at least 99% similar to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20. It is appreciated in the art that the descriptions they show for skilled in the art how to make and use a reference sequence (such as SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20) will also be sufficient to show an individual to make and use the described variants. Three commonly assigned patents describing nitrite-oxidizing bacteria, methods of using the bacteria and methods to detect bacteria, filed in the United States (see U.S. Patent Nos. 6,207,440, 6,265,206 and 6,268,154). These three patents describe a nucleotide sequence and any of the variants that have more than 96.1% homology to such a sequence. The publication of these patents demonstrates that the specifications that establish the particular sequences and that describe the particular variants allow an expert in the art to make and use the sequence and its variants described. Furthermore, it is common in the art that patents that describe nucleotide sequences also describe and claim variants of these sequences (see, for example, U.S. Patent Nos. 6,465,621, 6, 509, 170 and 6, 573, 066). The variants of the particular nucleotide sequences may be naturally occurring polymorphisms or alterations of synthetic sequences (see, for example, U.S. Patent No. 6,485,938). A great variety of modifications to the nucleotide sequences, natural or synthetic, are common and well known in the art, together with the methods for making the synthetic variants (see, for example, US Patent Nos. 6, 448, 044 and 6). , 509,170). Methods for comparing the similarity of two or more nucleotide sequences are well known in the art. Often two similar sequences are identified using computer programs such as BESTFIT and BLAST (see, for example, U.S. Patent No. 6,461,836). In addition, hybridization can be used to detect similarity between variant sequences and a reference sequence (see, for example, U.S. 6, 573, 066). Thus, one skilled in the art would be able to easily synthesize and identify the nucleotide sequences that are variants of a reference sequence using known techniques. Therefore, a specification describing a nucleotide sequence and its variants allows one skilled in the art to make and use this sequence and its variants.

EXAMPLES A series of tests and experiments were carried out to isolate, identify and show the efficacy of the bacterial strains reported herein. These involved a variety of bacterial culture techniques, biological molecular analysis of DNA extracted from culture samples, molecular biological analysis of bacterial strains and the application of concentrated cultures of bacterial strains to aquaria to measure their ability to control concentrations of bacteria. ammonia.

EXAMPLE 1 Culture of Bacteria Bacterial culture vessels (called reactors) were built and planted with accumulated bacterial biomass from the aquariums in operation. Each reactor received 4.95 L of a mineral salt solution (made in distilled water) containing 50 g of KH2P04, 50 g of K2HP04, 18.75 g of MgSO4-7H20, 1.25 g of CaCl2-2H20 and 1 g of FeS04-7H20. Air was provided so that the dissolved oxygen was equal to or greater than 7.5 mg / L, stirring was provided and the reactors were kept in a dark cabin at about 28 ° C. For the isolation and culture of the AOB strains of the present invention in salt water environments, synthetic sea salts (INSTANT OCEAN, Aquarium Systems Inc., Mentor, OH) were added until reaching a salt concentration of between 30 and 33 ppt . The concentrations of ammonia and nitrite were measured per day using flow injection analysis (FIA system, Tecator FIAStar 5010) while the pH was determined with an electrode (pH meter / ISE model 225 from Denver Instruments and was associated with an electrode pH / ATC). Nitrate and conductivity were measured periodically and the data were used to determine when water changes were required. The bacterial biomass was retained in the reactors during the water changes because the biomass is very flocular in nature. In this way, to decant 50% of the reactor volume by means of the appropriate sampling port, the biomass was sedimented by changing the air and the stirring mechanism for one hour. Additionally, the reactors were periodically purified to remove the biomass from the surfaces and in this way returning the biomass to the suspension. Microbiological samples were routinely taken for DNA extraction (by PCR) and cell fixation (by FISH) for further analysis.

EXAMPLE 2 Sampling and Extraction of Nucleic Acid For extraction of DNA, samples of appropriate biological filtration media were taken and resuspended in a cell lysis buffer (40 mM EDTA, 50 mM Tris-HCl, pH 8.3). Samples were stored at -20 ° C or -74 ° C until extraction. For processing, lysozyme was added to the samples to a final concentration of 10 mg / ml. After incubation at 37 ° C for 90 minutes, sodium dodecyl sulfate (SDS) was added at 20% to a final concentration of 1%. The samples were then subjected to four freeze / thaw cycles by the addition of proteinase K (standard concentration, 10 mg / ml) to a final concentration of 2 mg / ml and incubated at 70 ° C for 35 minutes. In some cases, additional proteinase K and SDS were added and the sample was incubated at 55 ° C for another 30 minutes. After cell lysis, the DNA was extracted using the Easy DNA extraction kit (Qiagen Inc., Santa Clarita, CA, hereinafter "Qiagen"). The DNA was eluted to a volume of 50 μ? and Hoechst type 33258 binding dye and fluorometry were quantified (DynaQuant 200, Hoefer Pharmacia Biotech Inc., San Francisco, CA).

EXAMPLE 3 Clones libraries of rRNA genes amplified by PCR Clones libraries of DNA extracts were derived from biomass samples taken from reactors and aquariums. The bacterial ribosomal RNA gene fragments of the bacteria represented by the sequences SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18 and SEQ ID NO: 20 , were amplified with the primers SD-Bact-0011-aS-17 (8f, GTT TGA TCC TGG CTC AG) (SEQ ID NO: 13) and 1 92r (eubacterial; GTT TAC CTT GTT ACG ACT T) (SEQ ID NO. : 14). The PCR conditions, the cycle parameters and the reaction components were as previously described (DeLong, E.F. 1992. Archaea in coastal marine environments, Proc.Nat.Acid Sci USA 89: 5685-5689). The PCR products were evaluated by agarose gel electrophoresis. The PCR fragments were cloned with a TA cloning kit (Invitrogen, Carlsbad, CA), as described in the manufacturer's instructions, after rinsing with the TE buffer and concentrating at 30 μ? with a CENTRICON concentrator (Amicon, Inc. Beverly, MA).

EXAMPLE 4 Sequencing and phylogenetic analysis. The 16S rDNA insert of each clone comprising the clone library was selected by restriction enzyme analysis (REA) using the restriction enzyme Hae III to ensure that the 16S rDNA insert could be amplified and determining if the 16S rDNA had a single REA pattern, then the plasmid of the clone containing the 16S rDNA insert of interest was partially sequenced. The amplified PCR 16S rDNA template of each clone selected for sequencing was cleaned using PCR Purification Kit catalog number 28142 (Qiagen). Sequencing was performed "using an automated LiCor 4000L DNA sequencer on the template that was sequenced by cycles with fluorescently labeled primers and the SEQUITHERM EXCEL II DNA sequencing kits (Epicenter Technologies, Madison, WI). three clones of the same REA pattern were partially sequenced to ensure they were identical, many clones were completely sequenced and analyzed phylogenetically by PAUP (phylogenetic analysis using Parsimony version 4.0b2a, DL Swoffrod) (analysis of input command values and distance analysis), ARB (an environment of programming elements (software) for the duration of the sequence, W. Ludwig and 0. Strunk) (phylogenetic tree) and Phylip (Phylogeny Inference Package J. Felsentein) (similarity matrix) The primers and probes for the clone of interest of the clone libraries were developed using the ARB probe design and the Equalization of probes as well as the subsequent manual alignment. The primers and probes were double-checked with BLAST (S.F. Altschul et al., 1990. Basic local alignment tool, J. Mol. Biol. 215: 403-410). The specificity of the primers was determined by using them on the DNA extracted from the clones and the pure cultures of the known bacteria. The specificity of the probes was tested using the pure cultures of the known bacteria and the samples of the reactors.

EXAMPLE 5 DGGE Analysis and Profile Formation For general eubacterial DGGE analysis, rDNA fragments were amplified using the forward 358f (eubacterial; CCT ACG GGA GGC AGC AG) (SEQ ID NO: 15) with a 40 bp GC clip over the 5 'end as described by Murray et al. (A. Murray et al 1996. Phylogenetic compositions of bacterioplankton from two California estuaries compared by denaturing gradient gel electrop oresis of 16S rDNA fragments, Appl.Environment Microbiol. 62: 2676-2680) and the reverse primer S - * - Univ. -0519-aA-18 (519r: GA TTA CCG CGG CKG CTG) (SEQ ID NO: 16). The specific DGO of AOB, the forward primer of 358f (SEQ ID NO: 15) with a 40 bp GC clip over the 5 'end was used with the reverse primer S - * - Ntros-0639-aA-20 (Nitroso4e: CAC TCT AGC YTT GTA GTT TC) (SEQ ID NO: 17). The PCR conditions were the same and were performed in a ROBOCYCLER GRADIENT 96 (Stratagene, La Jolla, CA) using the TAQ PCR center kit (Qiagen). The PCR conditions included a warm start procedure (80 ° C) and a test procedure. Initial denaturation at 94 ° C for 3 minutes was followed by a denaturation at 94 ° C for 1 minute, a proof anneal of 64 ° C at 55 ° C for 1 minute, 29 seconds (the annealing time during the test increased for 1.4 seconds per cycle) and the extension of the primer at 72 ° C for 56 seconds (the extension time was increased by 1.4 seconds per cycle). The initial temperature series of the above thermal cycle was repeated for 20 total cycles, followed by a final extension at 72 ° C for 5 minutes. Amplicons were examined by agarose gel electrophoresis. The DGGE was performed with a Bio-Rad D-GENE system (Bio-Rad Laboratories, Hercules, CA; subsequently "Bio-Rad"). The gels were 8.5% acrylamide / Bis using Bio-Rad reagents (D GENE electrophoresis reagent kit, Bio-Rad). The gel gradients were poured using Bio-Rad reagents (D GENE electrophoresis reagent kit, Bio-Rad) with a denaturation gradient of 20% to 60% (where 100% denaturation is a 40% formamide mixture deionized and urea 7M) and the Bio-Rad gradient release system (Model 475, Bio-Rad). All gels were run at 200 volts for 6 hours. The gels were visualized in one of two ways. By visualization and recovery of the discrete DNA bands, the gels were first stained for 10 minutes in 250 ml of TAE IX buffer in which 100 μ? of ethidium bromide (1 mg / ml), then washed for 10 minutes in TAE IX buffer. For documentation purposes, some gels were stained in Vistra Green (diluted 1: 10,000) (Molecular Dynamics, Sunnyvale, CA, hereinafter "Molecular Dynamics" (for 20 minutes, followed by a 20 minute wash in the TAE buffer IX and then were explored using a SI FLUORIMAGER (Molecular Dynamics) .The individual bands were removed from the DGGE gels using scalpels sterilized with alcohol.The extraction of the DNA from the gel followed by the methods of Ferris et al. (MJ Ferris et al. 1996. Denaturing gradient gel electrophoresis profiles of 16S rRNA-defined population inhabiting a hot spring microbial mat community, Appl. Environ Microbiol. 62: 340-346.) The suppressed band was placed in a sterile 2 ml screw-capped tube. with 500 μ? of sterile deionized water The tubes were filled in half with glass beads (cat No. 11079-101, Biospec Products Inc., Bartlesville, OK, hereinafter "Biospec") and placed in a agitator of mechanical beads (INI-BEADBEATER-8, Biospec) for 3 minutes at the highest calibration. The processed DNA was kept in the tubes at 4 ° C overnight. After storage overnight, the tubes were centrifuged at 3,200 X g for 8 minutes at 4 ° C to concentrate the gel fragments. The supernatant was transferred to a clean eppendorf tube. To verify the extraction efficiency, the supernatant was again amplified with the DGGE primers and retested by DGGE. An acceptable extraction was considered if it produced a single band in the DGGE analysis that co-migrated with the original DGGE band in the sample of the mixed population. The nucleotide sequence of the suppressed band was sequenced. by the methods described above using the fluorescently labeled primers.

EXAMPLE 6 Development of the oligonucleotide probe Oligonucleotide probes were designed that hybridize specifically with the 16S rRNA gene sequence isolated from closely related bacteria from the reactors in this study. A probe (SG-Nsspa-0149-aA-18) (SEQ ID NO: 5) is the target of two Nitrosospira-like bacteria derived from the reactor, which are represented by the sequences of SEQ ID NO: 1 and SEQ ID NO: 2 for the exclusion of other beta subdivision ammonia proteobacteria oxidants, including the sequences represented by SEQ ID NO: 3 and SEQ ID NO: 4, and also to the exclusion of Nitrosomonas aestuarii-like bacteria represented by SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20. A second probe (SG-Nsspa-0149-aA-19) (SEQ ID NO: 8) is the target of a bacteria similar to Nitrosospira derived from the reactor, which is represented by the sequence of SEQ ID NO: 3, to the exclusion of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4 and the other proteobacterial ammonia oxidants of the subdivision beta, and also to the exclusion of Nitrosomonas aestuarii-like bacteria represented by SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO; 20. Additional oligonucleotide probes were designed that hybridize specifically with the 16S rRNA gene sequence isolated from other reactor bacteria in this study. One probe, SG-Ntsms-0149-aA-18 (SEQ ID NO: 21), is the target of two bacteria similar to Nitrosomes aestuarii, which are represented by the sequences of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 for the exclusion of other AOB sequences represented by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, as well as sequences of bacteria similar to halophila. The pairings of the probes were initially selected using BLAST (SF Altschul et al., 1990. Basic local alignment tool, J. Mol. Biol. 215: 403-410) and CHECK_PROBE (BL Maidak, et al., 1994. The ribosomal datbase project. Nucleic Acids Res. 22: 3485-3487). The probes were synthesized by Operon Tech, Inc. (Alameda, CA). The nucleotide sequence and the position of the probes are shown in Table 10.

Table 10: Nucleotide sequences and positions of oligonucleotide probes and PCR primer groups for bacteria that oxidize ammonia.

Probe / primer Sequence base% Temp. of white Group (5'a 3 ') annealed formamide (° C) S-G-Nsspa-0149- CCC CCC TCT TCT 30 - SEQ ID NO: l & a-A-18 SEC ID NO: 2 (SEQ ID NO: 5) GGA TAC SG-Nsspa-0149- TCC CCC ACT CGA 20 - SEQ ID NO: 3 aA-19 (SEQ ID NO: 8) AGA TAC G SG-Ntsms-0149- CC CCC CT TCT 20 - SEQ ID NO: 18, aA- 18 GGA CAC SEC ID NO: 19 & (SEQ ID NO: 21) SEQ ID NO: 20 Probe / primer Sequence base% Temp. of White Group (5'a 3 'i annealed formamide (° C) Primer CGG AAC GTA TCC-5A SEQ ID NO 1 &front (SEQ ID NO: 6) AGA AGA SEQ ID NO 2 Reverse primer ATC TCT AGA AAA - (SEQ ID NO: 7) TTC GCT ATC primer GGA ACG TAT-56 SEC ID NO: 3 front (SEQ ID NO: 91 CTT CG Reverse primer CCA CCT CTC RGC - (SEQ ID NO: 10) GGG C TCA GAA AGA AAG-56 Primer SEQ ID NO: 4 Front (SEQ ID NO: 11) AAT CAT G Reverse Primer GTC TCC AYT AGA - (SEQ ID NO: 12) TTC CAA G Primer GTG ACT AAT AAT - 56 SEQ ID NO: 18, front (SEQ ID NO: 22) CAC AAC TTA SEQ ID NO: 19 &SEQ ID NO: 20 Reverse primer TTA TCT CTA AAA - (SEQ ID NO: 23) TAT TCG CT The severity for the probes (SEQ ID NO: 5, SEQ ID NO: 8 and SEQ ID NO: 21) was determined through a series of FISH experiments at different concentrations of formamide using the reactor biomass as a positive control for the bacterial sequences herein (SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20 ). The specificity of the probes was examined by testing against negative control cells from pure cultures of other beta subdivision ammonia oxidizing bacteria (Nitrosomas europaea, Nitrospira multiformis, Nitrosomas cryotolerans). In situ hybridization of the immobilized cells, fixed, it was carried out in a hybridization solution consisting of 0.9M NaCl, Tris / HCl 20 mM (pH 7.4), sodium dodecyl sulfate (SDS) 0.01%, 25 ng of oligonucleotide probe and different amounts of formamide. The slides were incubated in a humidity chamber equilibrated at 46 ° C for 90 to 120 min. The hybridization solution was rinsed with a preheated wash solution (48 ° C). The slides were then incubated in the wash solution for 15 minutes at 48 ° C. To achieve the same severity during the washing step, as in the hybridization step, the wash solution contained 20 mM Tris / HCl (pH 7.4), 0.01% SDS, 5 mM EDTA and NaCl. The concentration of NaCl was varied according to the percent of formamide used in the solution. For the 20% formamide, the NaCl concentration was 215 mM, for 30% it was 120 mM and for 40% the NaCl concentration was 46 mM. The cells were detected using an ANXIOSKOP 2 epifluorescence microscope (Cari Zeiss, Jena, Germany) equipped with filter groups for FITC / FLU03 and HQ CY3. The optimal severity was determined to be 30% formamide for the S-G-Nsspa-01 9-a-A18 probe. For the probe S-G-Nsspa-0149-a-A-19 the optimal severity was determined to be 20% formamide. Optimum severity was determined to be 20% formamide for the probe represented by SEQ ID NO: 21, and 20% formamide for the probe represented by SEQ ID NO: 24.

EXAMPLE 7 Development of the primer by PCR Two groups of the PCR primers were developed which specifically detect the Nítrosospira-like bacteria of the sequences of the present invention. A third group of PCR primers that specifically detects Nitrosomonas-like bacteria of the sequences of the present invention was developed. One group (SEQ ID NO: 6 and SEQ ID NO: 7) specifically detects Ni throsospira-like bacteria with the sequence SEQ ID NO: 1 and the sequence SEQ ID NO: 2 for the exclusion of other bacteria that oxidize ammonia (Table 11). The second group (SEQ ID NO: 9 and SEQ ID NO: 10) specifically detects Nitrosospira-like bacteria with the sequence SEQ ID NO: 3 for the exclusion of the other bacteria that oxidize the ammonia (Table 11). The third group (SEQ ID NO: 11 and SEQ ID NO: 12) specifically detects Nitrosomonas-like bacteria with the sequence SEQ ID NO: 4 for the exclusion of other bacteria that oxidize the ammonia (Table 11). A fourth group (SEQ ID NO: 22 and SEQ ID NO 23) specifically detects Nitrosomonas aestuarii-like bacteria with the sequences SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 for the exclusion of other bacteria that oxidize ammonia. The PCR conditions were described as above except that the annealing temperature was modified.

Table 11A: Results of the primer development specificity test by PCR and annealing temperature experiments.

Clone number or PCR of AOB Type A AOB PCR Type B AOB PCR Type C SEC bacterial species SEQ ID NO: 6 and SEQ ID NO: 9 and SEQ ID NO: 11 and SEQ ID NO: SEQ ID NO: 7 ID NO: 10 12 Temp. annealing 48 50 4 5 56 58 60 48 50 52 54 56 (° C) R7cl40 (Type A) + + + + - - - - - - R7cl87 (Type A) + + + + - - - - - R3c5 (Type B) - - - • - - - - Clone number or PCR of AOB Type A PCR of AOB Type B PCR of AOB Type C SEC bacterial species SEQ ID NO: 6 and SEQ ID NO: 9 and SEQ ID NO: 11 and SEQ ID NO: SEQ ID NO: 7 ID NO: 10 12 R5c20 (Type B) - - - - + + - - - - R3cl2 (Type C) - - - - - - - - + + + + R5c47 (Type C) - - - - - + + + + + N. europaea N. multiformis - - - - - - + + + - N. cryotolerants - - - - - ± + - P4c42 P4c31 BF16c57 Negative control Weak (±), Strong (+) or No signal (-) Table 11B: Results of the primer development specificity test by PCR and annealing temperature experiments.

Number of clone or species SEQ ID NO: 22 and of bacteria SEQ ID NO: 23 Temp. annealed 48 50 52 54 R7cl40 (Type A) R7cl87 (Type A) R3c5 (Type B) R5c20 (Type B) Number of clone or species SEQ ID NO: 22 and of bacteria SEQ ID NO: 23 R3cl2 (Type C) - - R5c47 (Type C) - - N. europaea N. multiformis N. cryotolerants P4c42 P4c31 BF16c57 Negative control Weak (+), Strong (+) or No signal (-). The specificity of each primer group was optimized by running a PCR experiment with each primer group using the temperature gradient mode of ROBOCYCLER from Stratagene. In this mode you can run a single experiment of all the reactions in up to 12 different annealing temperatures. Typically, the experiments were carried out from 4 to 6 different temperatures with an increase interval of 2 ° C. Each group of PCR primers was tested against the clone product with a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEC ID NO: 19 and SEQ ID NO: 20. The rDNA extracted from the pure cultures of Nitrosomonas europaea, Nitrosolobus multiformis and Nitrosomonas cryotolerans was also tested. Table 10 presents the PCR primer groups and the results of the optimum annealing temperature are shown in Table 11.

EXAMPLE 8 Similarity Analysis Thirteen libraries of clones were constructed from a number of freshwater and saltwater nitrification biomasses to determine the identity of the ammonia oxidant (s) sensitive to the oxidation of ammonia to nitrite. The details about the biomasses are presented in Table 12.

Table 12: Details regarding the receivers and aquariums from which the biomass was extracted and the clone libraries were built.

Library Details of nitrification biomass of clones Biofarm 16 This biomass was acquired from the BF16 carcass. Biofarm was dosed routinely at 300 mg / L / h of ammonia (NH3-N) for 6 hours per day. BC5 This biomass was kept in an aquarium (planted with a freshwater biofarm) and 5 mg / L of less ammonia was dosed every two or three days. The aquarium was not aerated. BC5 (2) Same as BC5 (previous). R3 This was seeded from an oxidation culture of enriched ammonia (approximately 1000 mg / L of NH3-N) that had been stored for 11 months. It grew at 5 mg / L of NH3-N and was aerated. R7 This was planted from Rl that had been planted with BC5. R1 and R7 were kept below 5 mg / L ammonia (NH3-N) and aerated.

Library Details of the nitrification biomass of clones R7BA6 This biomass was recovered from a test of Bacterial additive inoculated with biomass R7 R5 This biomass was derived from the biofarm feeding microfilter. It was exposed to extremely high concentrations of ammonia (> 500 mg / L of NH3-N). The reactor was operated at 30 mg / L ammonia (NH3-N) and aerated. R17 This biomass was derived from R7 but was fed at 30 mg / L for a period of three weeks before returning to 5 mg / L ammonia and aerated. R13 This biomass was derived from biomass BC5 but did not appear to have Nitroso-bacteria using the general AOB primers. It grew at 5 mg / L (NH3-N) and was aerated. R29 This biomass was acquired from the biofarm pit 5, which was a salt water biomass whose salinity remained between 30 and 35 ppt. This reactor was fed at 30 mg / L ammonia-nitrogen.

Library Details of nitrification biomass of P4 clones This reactor was seeded with 20 liters of biofarm pit material 15 which was a salt water biomass whose salinity was maintained between 30 and 35 ppt. This reactor was fed at 5 mg / L ammonia-nitrogen. SB7 This reactor was seeded with material from the biofarm stations 5 and 15 that were saltwater biomass whose salinity remained between 30 and 35 ppt. This reactor was fed at 5 mg / L ammonia-nitrogen. B7 This reactor was seeded with the material from biofarm stations 5 and 15 that were salt water biomass whose salinity was maintained between 30 and 35 ppt. This reactor was fed at 5 mg / L ammonia-nitrogen.

Data from the clone library show that there are three groups of bacteria that oxidize ammonia that exist in low ammonia feed reactors (eg, R3, R7). Not all three types of AOB were found to exist in each reactor considered. The three bacteria are represented by three groups of AOB clones - Type A of AOB (SEQ ID NO: 1) (and an Al subtype (SEQ ID NO: 2)) and type B of AOB (SEQ ID NO: 3). A fourth type was found in feed reactors elevated in ammonia-Type C of AOB (SEQ ID NO: 4). A similarity classification was performed for the four clonal sequences using RDP (aidak, BL, JR Cole, CT Parker, Jr, GM Garrity, N. Larsen, B. Li, TG Lilburn, MJ McCaughey, GJ Olsen, R. Overbeek, S. Pramanik, TM Schmidt, JM Tiedje and CR Woese, A new version of the RDP (Ribosomal Datbase Project), Nucleic Acids Res. 27: 171-173 (1999)) (Table 8). The similarity analysis showed that Type A of AOB (SEQ ID NO: 1) and Type Al (SEQ ID NO: 2) are 99.6% similar. This is in agreement with the 16S rDNA data which showed that there were 5 alignments in the 16Sr DNA between the type sequence for Type A (SEQ ID NO: 1) and the type sequence for Type Al (SEQ ID NO: 2). These similarity analyzes showed that the Type A and Al sequences are significantly different from the known AOBs from either Nítrosospira or Nitrosomonas casings (Table 13). This result is further confirmed by the analysis of the input command which shows that Type A of AOB (SEQ ID NO: 1) and Type Al (SEQ ID NO: 2) are grouped together in a group that is different from the Nítrosospira or Nitrosomonas casings (Fig. 1). In this way, the bacteria represented by Type A of AOB (SEQ ID NO: 1) and Type Al (SEQ ID NO: 2) are for the new species.

Table 13: Similarity fication for clones that oxidize ammonia isolated from reactors and aquariums % similarity to the rDNA of: AEN filter Similar Similar Similar Nitro- NitroNitro- NitroN ro- Nitro- a a a a sores sovibrio solofcus suspira sarcnas soaoocus Nitro-nitro-Nitro-Nitro-marira have nibberiensxs eurapaea rnabilis spira spira spira sarcrias formis Type? Type ftl Type B Type C Similar to Nitrosospira type A Similar to 0.996 Nitrosospira type ñl Sim lar to 0.944 0.942 Nitrosospira type B Similar to 0.934 0.932 0.925 Nitrosomonas type C Ni trosomonas 0.954 0.955 0.928 0.932 marine Ni rosovibrio 0.948 0.946 0.988 0. S26 0.932 tenuis Nitrosolobus 0.948 0.946 0.984 0.927 0.937 0.989 multiformis Nitrosospira 0.9 1 0.940 0.9"71 0.919 0.936 0.979 0.930 b iensis% similarity to the rDNA of: Source Similar to Nsm. sp. Nsm. sp. Nsm. Nsm. Nsm. marine Bact. Nsm. sp. of rDNA Nsm. Nsm. BF16c57 R7cl40 halrphi the aestuarü M96400 Marina NMS1 aest. aest. AF386746 AF386753 AF272413 AF272420 N03H AF272424 P4c31 P4c42 AF33820 Similar to | Nsm. aestuarü P4c31 Similar to 0.966 Nsm. aestuarü P4c42 Nsrr .. sp. 0.972 0.976 BF16c57 (AF386746) Nsm. sp. 0.981 0.981 0.983 P.7cl40 (AF386753) Nsm. 0.914 0.914 0.916 0.947 halophila (AF272413) Nsm. 0.983 0.977 1,000 0.983 0.916 (AE272420) Source Similar to Nsm. sp. Nsm. sp. Nsm. Man. Nsm. marine Bact. Nsm. sp. of rDNA Nsm. Nsm. ??? & 57 R7cl40 halcphüa aestuarü H96400 Marina NM51 aest. aest. AF386746 ÍF386753 AF272413 AF272420 N03W AF272424 P4c31 P4c42 SF33820 Nsm. 0.971 0.971 0.973 0.971 0.937 0.973 Marine (M96400) Bact. 0.994 0.994 0.996 0.980 0.944 0.996 0.974 tern N03W (AF33820) Ngn. sp. 0.951 0.959 0.982 0.979 0.915 0.982 0.989 0.981 MCI (AF272424) The similarity analysis for Type B of AOB (SEQ ID NO: 3) shows that this bacterium falls into the envelope of the Nitrosospira of AOB (Table 13). The analysis of the input command confirms this result (Fig. 1). However, the organism is sufficiently different from the nearest Nitrosospira AOB [Nitrosovibrio tenuis] that it can be considered as a new species. The similarity analysis for Type A of AOB (SEQ ID NO: 4) shows that this bacterium falls into the Nitrosomonas envelope of AOB (Table 13). The analysis of the input command confirms this result (Fig. 1). However, the organism is sufficiently different from the nearest Nitrosomonas AOB (Nitrosomonas europaea) that it can be considered as a new species.

Similarity analyzes were isolated for salt water tolerant AOBs, which are manifestations similar to Nitrosomonas aestuarii of clone P4c31 (SEQ ID NO: 19) which was 98.3% similar to Nitrosomonas aestuarii and clone P4C42 (SEQ ID NO: 18) ) is 97% similar to Nitrosomonas aestuarii. The phylogenetic analysis demonstrates the uniqueness of the sequences represented by clones P4C42 (SEQ ID NO: 18) and P4c31 (SEQ ID NO: 19) (Figure 6). The similarity classifications given in Table 13 are a guide for determining the uniqueness of one bacterial strain to another. There are no strict and fast rules regarding what percentage constitutes a new species. However, the Nitrosolobus multiformis and Nitrosovibrio tenuis that have a similarity classification of 0.989 are recognized by all the microbiological authorities as distinct species, such as Nitrosolobus multiformis and Nitrosospira briensis (similarity classification of 0.980). Since the similarity values of the bacterial strains reported in the present are not greater than for the pairs of species mentioned above, this is further evidence that the strains of the present are new and unique. Therefore, all of the clone data, PCR results, phylogenetic analyzes, DGGE data and similarity classification show that the bacterial strains reported here are unique and distinct from the known bacteria that oxidize the ammonia. In addition, it is expected that additional work in micro (or specialized) environments, such as those presented herein will result in the discovery of AOB related to the strains reported herein.

EXAMPLE 9 Analysis of Bacteria and Experimental Results Clone elements of AOB type A (SEQ ID NO: 1 and SEC) ID NO: 2) were found in the BF16 biomass (9% of the clone library) and the BC5 biomass (1-2% of the clone library) (Fig. 2). Biomass BC5 was used to seed the low concentration ammonia reactor (Rl), which was used to seed R7. The library of clones R7 generated from the biomass R7 containing only the AOB clones of type A (SEQ ID NO: 1 and SEQ ID NO: 2) (7% of the clone library) (Fig. 2). Here, AOB type A bacteria have been successfully subcultured from freshwater Biofarm to tank BC5 and then into reactor R7 by means of reactor Rl. This demonstrates the ability to successfully culture the bacteria and maintain a viable culture of AOB with the sequences herein. In addition, it demonstrates the ability to selectively enrich type A AOBs, since the percentage of this bacterium increased from 1-2% in the BC5 clone library to 7% in the R7 library. Apparently, the operation of the three systems (Biofarm 16, tank BC5 and reactor R7) would seem to be very different (see Table 12). However, there is a common group of physicochemical conditions that can explain the presence of type A AOBs in these systems. Although Biofarm receives high concentrations of ammonia initially, a period of time is allowed for the ammonia concentrations to fall to low levels (below 5 mg / L of NH3-N), thus allowing type A bacteria are retained in the system, which explains that it is able to grow a particular physiological niche at very low ammonia concentrations (< 5 mg / L NH3-N). Similarly, tank BC5 and reactor R7 were both fed and maintained at ammonia levels at or below 5 mg / L NH3-N. Type A AOB bacteria may be capable of existing at ammonia concentrations greater than 5 mg / L NH3-N, but it is evident that at higher ammonia concentrations are eliminated by other types of AOBs (ie Type B (SEC ID NO: 3) and / or Type C (SEQ ID NO: 4)) as evidenced by these types of AOB that are present and absent AOB type A, in reactors maintained at high ammonia concentrations (Table 14 ) (Fig. 2). The biomass of R7 did not flow particularly in the VI test of the microbial additives (BA6) and VII (BA7) (discussed below) and a biomass did not grow in the same way (R19) and with the same seeding (Rl) in the VIII test of bacterial additives (BA8) (R19). The AOBs of type A have been found in different reactors and in different test biomasses Post BA, both by PCR and FISH tests of AOB type A specific (Table 14) (Fig. 2). Therefore, these two AOB of Type A of newly discovered bacteria (SEQ ID No. 1) and Type Al (SEQ ID NO: 2) predominate in environments of low ammonia concentration, such as aquariums; and, when added to such an environment in a more purified state than in that occurring naturally, they can accelerate the establishment of ammonia oxidation in this environment (described below). Clonal elements of Type B were found in freshwater BioFarm biomasses (eg, BF 16-34% of the clone library) used to seed tanks BC (BC5). The AOB bacteria of type B were absent in the libraries of clones BC5 and R7, indicating that these AOBs may be more appropriate for the high ammonia conditions and the diet of a BioFarm (Fig. 2). Type B AOBs were also found in the R3 clone library (19% of the clone library) (Fig. 2). The history of the R3 reactor is that its biomass was initially enriched at high concentrations of ammonia (3000 mg / L of NH3-N), stored for 11 months and then matured in the reactor at low concentrations of ammonia (5 mg / L of NH3-N) for a prolonged period of time. During the initial growing period, probably ammonia concentrations decreased with time - thus motivating the growth of type C and / or B AOBs over type A AOBs. During the eleven months of storage, the ammonia would probably be removed, possibly motivating the maintenance of AOB type B bacteria in the system and survival of residual A-type AOBs that would have survived during the culture phase. Finally, during the maturation period in the reactor, the AOB type B bacteria would be able to be maintained, the AOBs of type A would be enriched and any residual type C that would have been originally selected for the original culture phase would be removed and would disappear. . In comparison, Biofarm biomass receives a relatively high concentration of ammonia for a set period and then gradually depletes it over time, creating a gradient of high to low ammonia concentrations (motivating the growth of type AOBs). B), often reaching zero, thus allowing a window for the growth of AOBs of type A - low concentrations of ammonia. This is a faster cycle (per day) than the cultivation phase of the R3 biomass, but by no means the least consistent with a change in conditions of high to low ammonia concentrations within the biomass. In this way, ammonia concentrations in Biofarm biomass motivate the enrichment of a range of AOB types as confirmed by the data from the clone library and the results of the DGGE tests. Type B AOBs have been found in a number of reactors and a number of Post BA test biomasses, both by specific B-type AOB PCR, DGGE and FISH. However, it has been found in as many post-bacterial additive tests or clone libraries as the AOB type A (Table 14). It appears that if the AOB was inoculated into a test, it was often recovered, whereas the AOBs of type B were recovered only in systems where AOBs of type A were not originally in the inoculum. Therefore, AOBs of type A were preferentially grown in the systems when they are present, but the AOBs of type B will satisfy when the AOBs of type A are present. While the AOBs of type A are the most important element of a community of successful AOB nitrification for low ammonia environments, such as the aquarium, are not the only AOB present. Another AOB, such as type B (SEQ ID NO: 3), may be necessary for the system to efficiently copy with ammonia fluctuation even for short periods of time (days). Type C AOBs are not desirable as an AOB in a bacterial additive for the low ammonia concentrations usually found in an aquarium. AOB type C bacteria were not found in the libraries of clones BF16, BC5 or R7 that are environments with a low concentration of ammonia, which indicates that they probably grew under conditions other than those found in these three environments (Fig. 2) . Type C bacteria were found in the libraries of clones R5, R3 and R17 (Fig. 2). The biomass of R5 grew consistently at high concentrations (30 mg / L of NH3-N) and its seed was of a very high concentration of ammonia (>500 mg / L of NH3-N), the biomass of R3 had originally grown to a high ammonia concentration before moving to a lower ammonia concentration (5 mg / L of NH3-N) and the biomass of R | 7 it moved from a low ammonia concentration (5 mg / L of NH3-N) to a high concentration (30 mg / L of NH3-N) and then returned again. The biomass of R5 had been enriched at high ammonia concentrations for a prolonged period even before being transferred to the R5 reactor, effectively excluding the growth of any of the AOB-type bacteria since the ammonia concentration never fell to low levels. the feeding microfilter. When the biomass was transferred to R5, the concentration of ammonia was reduced to lower levels, AOB type B was enriched and became the dominant AOB in this culture. Type C bacteria would represent the bacteria initially enriched in the microfilter and then remain in the R5 biomass when the feed was maintained at relatively high ammonia concentrations (30 mg / L NH3-N). The biomass of R3 had been allowed to grow initially at high ammonia concentrations, but over time the ammonia would become depleted. This regimen initially motivates the growth of type C AOBs (at higher ammonia concentrations) and type B AOBs (as ammonia was used). In addition, these pressures would not allow enrichment of type A AOBs that are dependent on consistently low ammonia levels. During the operation of the R3 reactor at lower ammonia concentrations, the AOB type C bacteria would be enriched and the type B bacteria would still survive, but since the AOB type A bacteria were originally minimized at the initial enrichment, there would be very few to take advantage of the new conditions inside the reactor. Therefore, AOB type B would be expected to be the dominant AOB in this environment. The biomass of R17 usually shows that it is not necessary to do to cultivate the AOBs of type A and / or B. The biomass of R17 was derived from the biomass of R7, but was cultivated for 3 weeks at high concentrations of ammonia (30 mg / L of NH3-N) to observe if a change in the microbial community would occur. There was no change and the type C AOBs became dominant, as demonstrated by the results of the FISH, PCR and DGGE experiments. In addition, the change was irreversible. After moving the biomass back to an environment of low ammonia concentration (5 mg / L of NH3-N), the AOB of type C remained as the dominant AOB, while the AOBs of type A and type B could not be detected either by FISH or DGGE. This suggests that during the three-week period the AOBs of type A and B were excluded from the biomass of R17. The biomass of R17 was poorly in the subsequent BA VIII test, suggesting that type C AOBs are not the correct type of AOB required for an effective bacterial additive to be used in the relatively low ammonia environment of an aquarium. This conclusion is also based on the results of bacterial additive tests that showed that commercial bacterial mixtures containing Nitrosomonas cover AOBs are not effective in accelerating the establishment of nitrification in aquariums (described below). Type C bacteria are closely related in a phylogenetic manner to bacteria found in wastewater treatment plants that also receive ammonia concentrations of approximately 30 mg / L NH3-N (similar to R5). The sets of PCR primers described herein were used to detect the presence or absence of the AOB strains reported herein, in a variety of environments. Environments include mixtures of the pre-bacterial additive test, aquarium filters of the bacterial additive test, and commercial mixtures of nitration bacteria manufactured and sold by other companies. further, the DNA extracted from the pure culture of another AOB was tested. Similar results from these experiments are summarized in Table 14. The data shows that the PCR primer sets are specific for the bacterial strain reported herein, and allow each strain to be detected exclusively from the other strains. In addition, pure cultures of known AOB are not amplified with any of the PCR primer groups reported herein. This shows that the bacteria reported herein can be distinguished from the known AOB. The data also shows that commercial additives, currently on the market, would be expected to fail to accelerate the establishment of nitrification in newly established aquariums, because these additives do not contain the correct bacteria species (detailed further in the next section).

EXAMPLE 10 Inspection by gel electrophoresis of the denatured gradient of the clones and reactors. The novelty of the different bacterial strains reported herein is further demonstrated by the results of the denatured gradient gel electrophoresis (DGGE) test. Figure 3 shows that the DGGE results for two representative clones for each of AOB type A (SEQ ID NO: 1), AOB of type Al (SEQ ID NO: 2), AOB of type B (SEQ ID NO: 3) ) and type AOB (SEQ ID NO: 4) in a general eubacterial DGGE. The bacterial sequence of each type of AOB described herein, is denatured at a different position in the gel. This is indicative of uniqueness and provides other means by which each type of AOB is distinguished from each other and also from the known AOB. None of the bacterial sequences observed previously co-migrated with Nitrosomonas europaea, Nitrospira multiformis or Nitrosomonas cryotolerans (Fig. 3). In addition, the DGGE analysis of the biomass extracted from the different reactors confirmed the results of the PCR and FISH test. Figure 7 presents the DGGE results that further distinguish the different AOB strains of the present invention. The bands representing the P4c42 and P4c31 clones similar to Nitrosomonas aestuarii (SEQ ID NO: 18 and SEQ ID NO: 19) do not co-migrate with the bands representing the AOB strains of AOB type A (SEQ ID NO: 1) ), AOB of type B (SEQ ID NO: 3), AOB of type C (SEQ ID NO: 4) and clone BF16c57 similar to Nitrosomonas aestuari (SEQ ID NO: 20). None of these strains co-migrate with Nitrosomonas europaea, Nitrospira multiformis or Nitrosomonas cryotolerans (Fig. 7).

EXAMPLE 11 Bacterial additive test A series of experiments were carried out to determine the efficacy of different bacterial mixtures containing the bacterial strains of the present invention compared to: (1) control aquariums that did not receive a mixture, (2) aquariums which were inoculated with bacterial mixtures for use in aquariums of tropical fish and (3) mixtures of bacteria preserved or stored from the bacterial strains of the present invention. The effectiveness of a mixture is demonstrated by showing that the bacterial strains that oxidize the ammonia of the present invention oxidize the ammonia in aquariums and, furthermore, that when combined with other bacterial strains (for example, bacteria that oxidize nitrite), the bacteria accelerate the establishment of nitrification in aquariums. The establishment of nitrification can be measured in at least three different ways. The first is counting the number of days it takes to establish a new aquarium so that the concentrations of ammonia and nitrite in the aquarium water reach concentrations close to 0 mg / L. In a new establishment of a freshwater aquarium, it usually takes approximately 14 days for the ammonia concentration to reach 0 mg / L and approximately 30 to 35 days for the nitrite to reach 0 mg / L. A second way to measure the beneficial action of the addition of bacterial nitrification strains to aquariums is to compare the maximum concentration of ammonia or nitrite reached before the concentration falls to 0 mg / L. If the maximum concentration of ammonia or nitrite reached in the aquariums in which the nitrification bacteria were added is significantly lower than the maximum concentration reached in the control aquariums, then a degree of effectiveness is demonstrated. A third way to evaluate the effectiveness of the nitrification bacterial strains and the mixtures that incorporate them is to combine the first two methods to form a toxicity exposure curve. This type of curve counts the duration (time in days) and the degree / intensity of the exposure. In accordance with the embodiments of the present invention, this curve is generated by plotting the concentration of the toxin over time. The area of the curve can then be determined for each treatment and the toxin by standard computer methods (for example, integrating mathematically the curve). The treatments of each test are then compared with each other and with the control of the same test. The area of the control curve can be given an arbitrary value of 1 and then the other areas can be calculated as a relation to the control area. Thus, if the value of a treatment is greater than 1 it is considered more effective than control, while a value less than 1 suggests that it is less effective than control and may have inhibited the establishment of nitrification.

EXAMPLE 12 Bacterial Additive Test VI The objective of this test was to evaluate the ability of four bacterial mixtures, including the bacterial strains of the present invention, to accelerate the establishment of nitrification in freshwater aquariums. The test was also performed to compare an ability to establish nitrification between the bacterial strains of the present invention and the control aquaria that did not receive a bacterial inoculation of any kind. Twenty-seven 37.854-liter (10-gallon) aquariums and twenty-seven tank-style energy filters on the back of Penguin 170B (Marineland Aquarium Products) were sterilized, rinsed thoroughly and allowed to air dry. Each aquarium was then filled with 4,536 kilograms (10 lbs) of rinsed aquarium gravel (RMC Lonestar # 3) and the filter was installed. The aquariums then received 35 1 of tap water from the city that had been filtered through activated carbon. After changing the filters, the water level in each aquarium was marked, so that it could be completely filled with deionized water (DI) to compensate for any water loss due to evaporation and sampling. The filters worked all night before the addition of the bacterial additives and the fish. On day 0 of the test, the aquariums were completely filled with DI water and a sample of baseline water was taken for analysis. Coal cartouches (Marineland Aquarium Products, part No. PA 0133) were rinsed with tap water and placed in each filter. New BIOWHEELS (Marineland Aquarium Products, part number PR 1935B) were placed in each filter. After thirty minutes, each tank was inoculated with its designated bacterial additive, or, if the tank was a control aquarium, it was not dosed with a bacterial mixture. Thirty minutes later, the experimental bacterial additives were added, a second group of water samples was taken for the analysis. Five pink barbels (Puntius conchonius) and one giant danio [Danio aequpinnatus] were added to each tank. The fish were then fed with approximately 0.4 grams of tropical fish flakes divided into two feeds per day (at approximately 9:00 a.m. and: 30 p.m.). The water samples were collected and analyzed for the test per day with respect to pH, ammonia, nitrite and conductivity. On Monday, Wednesday and Friday the water was tested for nitrate and turbidity. Anions and cations were periodically measured. The pH measurements were made with a model 225 pH / ion meter from Denver Instruments equipped with a pH combination electrode from Denver Instruments. A FIAstar 5010 Tester Analyzer was used to measure ammonia, nitrite and nitrate (ie, as nitrogen) using the methods described in the Tecator application recommendations. The cations (sodium, ammonia-nitrogen, potassium, magnesium and calcium) were analyzed using a Dionex DX500 System with a 4 mm CS15 analytical column. The specific conductance was measured directly in each tank at approximately 12:30 p.m. daily using a system of salinity, conductivity and temperature fastened with the hand model 30 of YSI. The turbidity data was determined with a turbidity meter DRT-100 (HF Scientific). Four bacterial mixtures were used in this test, and two dosage levels were implemented within each treatment: either 30 ml or 100 ml of a mixture per aquarium. Three replicates of each mixture / dose were tested, along with three control aquariums that did not receive a bacterial mixture; totaling 27 test aquariums (ie, (4x2x3) +3 = 27). The conditions for the different bacterial mixtures were as follows: 1) BC5 - a bacterial mixture that had been in culture for 553 days before the test. A positive result with this mixture would demonstrate the long-term viability of the bacteria under culture conditions and the convenience of culture techniques; 2) Rtr3 - a bacterial mixture that had been bottled and stored in the dark for 118 days before the test. A positive result with this mixture would show that the storage method is valid and the mixture would maintain its viability for at least 119 days of storage; 3) Rtr4 - a bacterial mixture that had been bottled and stored in the dark for 118 days before the test. A positive result with this mixture would show that the storage method is valid and the mixture would maintain its viability for at least 119 days of storage; 4) Rtr7 - a bacterial mixture that had grown from an inoculum of BC5. A positive result with this mixture would show that the bacterial consortium can be grown in the mixture for successive generations and maintain its viability. The test continued for 23 days, at the end of which the concentrations of ammonia and nitrite in the aquariums were virtually 0 mg / L. There were no significant differences between the highest ammonia and nitrite concentrations for the different aquariums, as well as the length of time necessary for the aquariums to reach a concentration of 0 mg / L. Other differences between the bacterial mixtures are shown in Table 11.

Table 14: Detection of bacteria that oxidize ammonia AOB PCR AOB PCR AOB PCR Type A Type B Type C R7cl40 (Type A) +++ - - R7cl87 (Type A) +++ - - R3c5 (Type B) - +++ - R5c20 (Type B) - +++ - R3cl2 (Type C) - - +++ R5c47 (Type C) - +++ PCR AOB PCR AOB PCR model of type A A type B AOB N. europaea - - - N. multiformis - - - N- cryotolerans - - - BC5 Pre BA 6 - - - BC5 Post BA 6 + - - R3 Pre BA6 + + + - R3 Post BA6 + +/- - R4 Pre BA6 + + - R4 Post BA 6 ++ - - R5 Pre BA 7 - ++ ++ R5 Post BA7 - - - R7 Pre BA6 ++ + - R7 Post BA6 ++ - - R7 Pre BA7 + +/- - R7 Post BA7 ++ - - Cycle - - - Fritzyme - - - Stresszyme - - - Cryst Cir Nitrifier - - - Cryst Cir Bio Ciar L - - - Cryst Cir Bio Ciar S - - - PCR AOB PCR AOB PCR AOB type A type B type C Acqmar Phospaht - - - Trop Sci Sludge - - - Trop Sci Rapid Act - - - +++ very strong presence, clearly indicates high amount of target organism ++ strong presence, indicates significant signal detection + clear presence, detected signal +/- possible presence, weak signal but previous background - no presence / signal detected Table 15: Results of the VI Test of bacterial additives.

Mix Time to < 0.50 Maximum bacterial concentration mg / L (days) average (mg / L-N) Ammonia Nitrite Ammonia Nitrite Rtr7 - 100 mi 6 7 1.1 1.4 Rtr3 - 100 mi 7 11 1.9 3.4 Rtr7 - 30 mi 7 10 1.9 3.7 BC5 - 100 mi 8 18 2.4 4.5 Mix Time a < 0.50 Bacterial concentration mg / L (days) maximum average (mg / LN) Ammonia Nitrite Ammonia Nitrite Rtr4 - 100 ml 8 8 2.7 0.6 Rtr3 - 30 mi 9 15 3.1 5.9 Rtr4 - 30 mi 9 10 2.9 2.1 BC5 - 30 mi 10 21 4.1 8.9 Control 12 23 4.9 13.4 Figure 4 shows the average concentrations of ammonia and nitrite during the test period for the four mixtures along with the controls. For the BC5 mixture, the ammonia reached 0 mg / L on day 8 for the aquariums dosed with 100 ml of the BC5 mixture on day 10 for aquaria dosed with 30 ml of the BC5 mixture. The concentration of ammonia in the control aquariums did not reach 0 mg / L until day 12. The mean concentration of ammonia highest reached for the control aquariums was 4.9 mg / L. However, for aquariums dosed with 30 ml of the BC5 bacterial mixture, the highest average concentration of ammonia was 4.1 mg / L, whereas in aquariums dosed with 100 ml of the BC5 bacterial mixture, the average concentration of ammonia highest was 2.4 mg / L.

In this way, the addition of the BC5 bacterial mixture to the newly established aquariums resulted in less exposure of ammonia to the fish. The values of the area of the ammonia exposure curve for the aquariums dosed with 30 ml or 100 ml of the mixture BC5 were 67% and 37% of the value of the area of the control aquarium curve, respectively (Table 16); 1.5 and 2.7 times less exposure to ammonia, respectively, for fish in treatment aquaria.

Table 16: Toxicity exposure data for the VI test of bacterial additives.

The nitrite concentrations reached 0 mg / L on day 18 in aquariums dosed with 100 ml of BC5 mixture, on day 21 in aquariums dosed with 30 ml of mixture BC5 and on day 23 in control aquaria. The control aquariums reached an average maximum nitrite control of 13.4 mg / L, while aquariums dosed with 30 ml of the BC5 mixture had an average maximum nitrite concentration of 8.9 mg / L and those dosed with 100 ml of the mixture BC5 had a maximum nitrite concentration of only 4.5 mg / L (Table 15, Fig. 4). The values of the area of the nitrite exposure curve for the aquariums dosed with 30 ml or 100 ml of the BC5 mixture were 75% and 33% of the area value of the control aquarium curve, respectively (Table 16); 1.3 and 3.1 times less exposure to nitrite, respectively, for fish in treatment aquaria. Aquariums that include the bacterial mixture Rtr3, which had been stored for 118 days, established nitrification faster than control aquariums did. The mean maximum ammonia concentration for aquariums dosed with 30 ml or 100 ml of the Rtr3 mixture was 3.1 and 1.9 mg / L, respectively (Table 15). In contrast, the control tanks showed an average maximum ammonia concentration of 4.9 mg / L. Control aquaria reached an ammonia concentration of 0 mg / L after 12 days, while aquariums dosed with 30 ml or 100 ml of the Rtr3 bacterial mixture took only 9 and 7 days to reach 0 mg / L, respectively ( Table 15). The mean maximum nitrite concentration was 13.4 mg / L in the control aquariums, while the mean maximum nitrite concentration in aquariums dosed with 30 ml or 100 ml of the Rtr3 bacterial mixture was only 5.9 mg / L and 3.4 mg / L, respectively. Control aquariums reached a nitrite concentration of 0 mg / L in 23 days, while aquariums dosed with 30 ml or 100 ml of the Rtr3 bacterial mixture reached 0 mg / L after only 15 and 11 days, respectively (Table fifteen) . The values of the area of the ammonia exposure curve for aquariums dosed with 30 ml or 100 ml of the Rtr3 mixture were 45% and 26% of the area value of the control aquarium curve, respectively (Table 15); 2.2 and 3.9 times less exposure to ammonia, respectively, for fish in treatment aquaria. The values of the area of the nitrite exposure curve for aquariums dosed with 30 ml or 100 ml of the Rtr3 mixture were 28% and 11% of the area value of the control aquarium curve, respectively (Table 15); 3.6 and 8.8 times less exposure to nitrite, respectively, for fish in treatment aquaria.

For aquariums dosed with the Rtr4 mixture, the mean maximum ammonia concentration was 2.9 mg / L and 2.7 mg / L, respectively, for a dosing volume of 30 ml and 100 ml (Table 15), while the aquaria of control reached an average maximum ammonia concentration of 4.9 mg / L. The values of the area of the ammonia exposure curve for aquariums dosed with 30 ml or 100 ml of the Rtr4 mixture were 45% and 38% of the area value of the control aquarium curve, respectively. These values show that the addition of the mixture resulted in 2.2 and 2.7 times less exposure to ammonia, respectively, for the fish in the treatment aquaria when compared to the control aquariums (Table 15). The aquariums dosed with 30 ml of the Rtr4 mixture completed the nitrification cycle in 10 days, while the nitrification was established in 8 days for the tanks dosed with 100 ml of the Rtr4 mixture (Table 15). Nitrification was established in 23 days in the control aquariums. The average maximum nitrite concentration for aquariums dosed with 30 ml or 100 ml of the Rtr4 bacterial mixture was 2.1 mg / L and 0.6 mg / L, respectively. The control aquariums had an average maximum nitrite concentration of 13.4 mg / L. The values of the area of the nitrite exposure curve for aquariums dosed with 30 ml or 100 ml of the Rtr4 mixture were 5% and 2% of the area value of the control aquarium curve, respectively; 20.2 and 60.9 times less exposure to nitrite, respectively, for fish in treatment aquaria (Fig. 4, Table 15). The Rtr7 bacterial mixture, which was a subculture of the BC5 mixture, demonstrated a significantly faster establishment of nitrification when compared to the control aquariums. The control aquariums took 12 days to reach an ammonia concentration of 0 mg / L, while aquariums dosed with 30 ml or 100 ml of the bacterial mixture of Rtr7 took only 7 and 6 days, respectively (Table 15). The mean maximum ammonia concentration for aquariums dosed with 30 ml or 100 ml of the R7 mixture was 1.9 mg / L and 1.1 mg / L, respectively. This is contrary to the control aquariums that had an average maximum ammonia concentration of 4.9 mg / L (Table 15). The nitrite concentration reached an average maximum concentration of 13.4 mg / L in the control aquariums, while in aquariums dosed with 30 ml or 100 ml of the Rtr7 bacterial mixture, the mean maximum nitrite concentration was only 3.7 mg / L and 1.4 mg / L, respectively (Table 15). Control aquaria reached a nitrite concentration of 0 mg / L in 23 days, while aquaria dosed with 30 ml or 100 ml took only 10 and 7 days, respectively, to reach 0 mg / L (Table 15, Fig. 4) . The values of the area of the ammonia exposure curve for aquaria dosed with 30 ml or 100 ml of the Rtr7 mixture were 28% and 17% of the area value of the control aquarium curve, respectively (Table 15); 3.6 and 5.7 times less exposure to ammonia, respectively, for fish in treatment aquaria. The values of the area of the nitrite exposure curve for aquariums dosed with 30 ml or 100 ml of the Rtr7 mixture were 10% and 4% of the area value of the control aquarium curve, respectively; 5.7 and 25.5 times less exposure to nitrite, respectively, for fish in treatment aquaria (Table 15). In summary, the test data show that the different bacterial mixtures of the present invention accelerate the establishment of nitrification in aquariums. The use of these mixtures in aquariums significantly reduced the degree of ammonia and nitrite exposure to fish. The results further demonstrate that a mixture can be kept viable for a prolonged period of time (eg BC5), that the mixture can be stored for several months (eg Rtr 3 and Rtr 4) and that successive generations of the mixture retain their nitrification capacity (for example Rtr 7).

EXAMPLE 13 Test VII of Bacterial Additive The objective of this test was to evaluate two mixtures of bacterial strains of the present invention as they were implemented in a "real world" installation, while comparing their performance with that of commercial bacterial mixtures. In general, a new owner of an aquarium first buys the necessary equipment to install an aquarium capable of maintaining aquatic life. The equipment can include the aquarium itself, decorations, a heater and a filter and a water conditioner. The aquarium is then assembled and filled with water, the filters are turned on, the heater is adjusted to the appropriate water temperature and the water conditioner is added to remove the chlorine. At this point, fish are usually added, but there may be insufficient populations of bacteria that oxidize ammonia and nitrite present to maintain the ammonia and nitrite concentrations in the aquarium at biologically safe (ie, non-toxic) concentrations (e.g. , below 0.5 mg / LN). Therefore, the newly installed aquarium will exhibit what is commonly referred to as "new tank syndrome" (ie, high concentrations of ammonia and nitrite in the first weeks after installing a new aquarium when an insufficient population of bacteria is present). nitrification to maintain safe concentrations of ammonia and nitrite).

To resolve at least partially the syndrome of the new tank, a bottled mixture of microorganisms or a mixture of enzymes (ie the bacterial mixture) can be purchased and introduced into the new aquarium to accelerate, or in some cases, eliminate the syndrome of the new tank. . In theory, the introduction of the bottled mixture should result in comparatively lower ammonia and nitrite concentrations in an aquarium during its initial weeks than in the absence of this mixture. Also, it should take less time for the ammonia and nitrite concentrations to reach 0 mg / L. Thirty-three 37.854-liter (10-gallon) aquariums and thirty-three style energy filters hung on the back of Penguin 170B (Marineland Aquarium Products) were sterilized, rinsed thoroughly and allowed to air dry. Each aquarium was then filled with 4,536 kilograms (10 lbs) of rinsed aquarium gravel (RMC Lonestar # 3) and the filter was installed in the back. The aquariums were then filled with 35 L of water from the city that had been pre-filtered through activated charcoal and the water level was marked in each aquarium. This mark was used as a guide to indicate when the aquarium water needed to be completely filled to compensate for the water lost due to evaporation or sampling. Deionized water was used to completely fill the aquariums. The filters were allowed to run overnight before the addition of bacterial additives and fish. On the first day of the test, the aquariums were filled completely with deionized water to consider the water and a water sample from the baseline. Carbon cartridges (Marineland Aquarium Products, part No. PA 0133) were rinsed with tap water and placed in each filter. New BIOWHEELS (Marineland Aquarium Products, part number PR 1935B) were placed in each filter. After thirty minutes, each tank was inoculated with its designated bacterial additive. The dosages were as described in Table 17. Thirty minutes later, the bacterial additives were added, a second group of water samples were extracted for analysis. Six different barbels were added to each tank [. { Puntius conchonius) Pink barbels; (Puntius tetrazona), albino tiger barbels (Albino Tiger) and tiger chins (Tiger Barbs)]. The fish in each aquarium were fed twice a day (at approximately 9:00 a.m. and again at 4:30 p.m.) with a total of 0.4 grams of tropical fish flakes per day. Water samples were collected and analyzed for the test per day with respect to pH, ammonia, nitrite and conductivity. On Monday, Wednesday and Friday the water was tested for nitrate and turbidity. Anions and cations were periodically measured. The pH measurements were made with a model 225 pH / ion meter from Denver Instruments equipped with a pH combination electrode from Denver Instruments. A FIAstar 5010 Tester Analyzer was used to measure ammonia, nitrite and nitrate (as nitrogen) using the methods described in the Tecator application recommendations. The cations (sodium, ammonia-nitrogen, potassium, magnesium and calcium) were analyzed using a Dionex DX500 System with a 4 mm CS15 analytical column. The specific conductance in each tank was measured directly at approximately 12:30 p.m. daily using a system of salinity, conductivity and temperature fastened with the hand model 30 of YSI. The turbidity data was determined with a turbidity meter DRT-100 (HF Scientific). Two formulations containing bacterial strains of the present invention were tested together with four commercially available bacterial mixtures. On the first day of testing, 100 ml of the test formulation (Rtr5) was added to each of the four aquariums and 100 ml of the second formulation (Rtr7) was added to four other aquariums. The commercially available bacterial mixtures were dosed according to the manufacturer's instructions, for the treatments of BIOZYME, CYCLE, FRITZ-ZYME NO. 7 and STRESS ZYME. In addition, each of these commercially available bacterial mixtures was also tested at three times the recommended dosage level (Table 17). There were four replica aquariums per treatment / dosage combination for a total of 33 aquariums (ie, (((4x3) x2) + (2x3) +3) = 33). The trends of ammonia and nitrite for the treatments and control for test VII of the bacterial additives are shown in Figure 5. For clarity of presentation, each of the commercially available bacterial mixtures tested is presented with the control and the two mixtures of test containing the bacterial strain of the present invention. The scale of each graph is the same, so that comparisons can easily be made between all treatments. The data show that the Rtr5 and Rtr7 mixtures, which contain the bacterial strains of the present invention, significantly decreased the time needed to establish nitrification in newly installed aquariums compared to aquariums that were not dosed (ie, controls) or who received a commercially available bacterial mixture. In addition, the concentrations of ammonia and nitrite reached in the aquariums that were dosed with the bacterial mixture Rtr5 and Rtr7 were significantly lower than in all other treatments (Fig. 5).; Table Table 18: Results of test VII of bacterial additives The Rtr5 and Rtr7 mixtures established nitrification in newly installed aquariums significantly faster than commercial mixtures and untreated aquariums. The complete nitrification was established in 8 days with the mixture of Rtr7 and in 10 days with the mixture of Rtr5 (Table 18). The treatments closest to these were FRITZ-ZYME at its normal dosage level, CYCLE at three times its normal dosage level and STRESS ZYME at its normal dosage level; each of which took 22 days (Table 18). Therefore, the mixtures of Rtr5 and Rtr7 were 2.2 to 2.8 times faster for the establishment of nitrification than these other mixtures. The difference in the maximum concentration of ammonia or nitrite achieved for the different mixtures and the control were also significantly different (Table 18). The mean maximum concentration (N = 3) of 1.5 mg / L reached during the test for the Rtr5 mixture was 4.8 times lower than the FRITZ-ZYME (mean 7.2 mg / L, N = 3), dosed at its normal level, which was the closest commercially available mixture (Table 18). The mean maximum nitrite concentration for the Rtr5 mixture was 0.9 mg / L. Again, FRITZ-ZYME dosed at its normal level was the closest commercially available mixture with an average maximum nitrite concentration of 3.1 mg / L. Therefore, the Rtr5 mixture was 3.4 times more effective in establishing nitrification than the currently available commercial blends tested. The Rtr7 mixture exhibited the same tendency as the Rtr5 mixture in that aquaria dosed with this mixture exhibited significantly lower concentrations of ammonia-nitrogen and nitrite-nitrogen than aquariums dosed with commercially available bacterial mixtures (Table 18). The Rtr7 mixture had average maximum ammonia and nitrite concentrations of 2.8 mg / L and 1.3 mg / L, respectively, These were 2.6 and 2.4 times lower, respectively, than the closest commercially available bacterial mixture (FRITZ-ZYME, dosed at its normal level) (Fig. 5, Table 18). In terms of the exposure curves, the bacterial mixtures of Rtr5 and Rtr7, which incorporate the bacterial strains of the present invention, developed significantly better than the commercially available mixtures (Table 19). In particular, Rtr7 developed better than any mixture with the fish exposed to only 13% of the ammonia and 5% of the control nitrite. Rtr5 was almost as effective, with ammonia exposure at 14% of control levels and nitrite exposure at 9% of control (Table 19). These results mean that fish in aquariums receiving Rtr7 or Rtr5 are exposed from 7.3 to 7.6 times less ammonia and 11.6 to 19.6 times less nitrite than fish in the control aquariums. After Rtr7 and Rtr5, the following best mixtures reduced the exposure of ammonia and nitrite to only 50% when compared with the controls (Table 19).

Table 19: Toxicity exposure data for test VII of bacterial additives.

EXAMPLE 14 Test VIII of bacterial additive The objective of this test was to evaluate different mixtures of AOB strains of the present invention since they can be implemented in a "real world" installation. The performance of these mixtures of the present invention was compared with other AOB strains of the present invention, as well as with commercially available bacterial mixtures which claim that they are suitable for use in freshwater and saltwater aquariums. For this test, fifteen 37,854-liter (10-gallon) aquariums and fifteen energy-style filters hung on the back of Penguin 170B (Marineland Aquarium Products) were sterilized, rinsed thoroughly and allowed to air dry. The next day, each tank was filled with 4,536 kilograms (10 lbs) of coral Crushed Tideline # 5 and was equipped with an esterified energy filter (PF 0170B) and rinsed with carbon cartridge. Each tank was filled with 35 L of artificial seawater. The seawater was a combination of a mixture of Tropic Marine salt and post GAC water at a salinity of 30 ppt. The filters were allowed to run overnight before the addition of the bacterial additives and the fish. The next morning the tanks were filled completely with ultrapure water to compensate for the evaporation and the water mixtures taken. Each tank was dosed with a bacterial treatment, however, no bacterial mixture was added to the control group. There were four treatments for this test: Reactor 3 included the AOB strains of the present invention represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 / reactor 29 included strains of AOB of the present invention represented by SEQ ID NO: 18, SEQ ID NO: 19 and the sequences of the two strains similar to halophila; CYCLE (a bacterial mixture commercially available for use in fresh or salt water); and STRESS ZYME (another bacterial mixture commercially available for use in fresh or salt water). Each treatment had three replications. The aquariums receiving the treatments of Reactor 3 and Reactor 29 were dosed with 100 ml of any mixture once on the first day of the test. Aquariums receiving CYCLE or STRESS ZYME treatments were dosed with 10 ml of any treatment on the first day of the test, an additional 10 ml on day 7 of the test and an additional 5 ml every 7 days after the duration Of the test. Four damsels were added to each tank. { Pomacentrus spp. ) varied on the first day of the test and were fed twice a day. Water samples were collected and analyzed per day for pH, ammonia, nitrite and conductivity. On Monday, Wednesday and Friday, water was tested for nitrite and turbidity. The pH measurements were made with a model 225 pH / ion meter from Denver Instruments equipped with a pH combination electrode from Denver Instruments. A Tecator FIAstar 5010 analyzer was used to measure ammonia, nitrite and nitrate (as nitrogen) using the methods described in the Tecator application recommendations. Salinity was measured directly in each tank per day using a salinity, conductivity and temperature system fastened with the YSI model 30 hand. The turbidity data were determined with a turbidity meter DRT-100 (HF Scientific). The average ammonia concentrations for the four treatments and control are depicted in Figure 8. The treatment reactor 29, which consisted of strains of AOB represented by SEQ ID NO: 18, SEQ ID NO: 19 and two N-like strains. halophila, oxidized the ammonia considerably faster than the other treatments. The mean maximum ammonia concentration of Treatment Reactor 29 was also significantly lower than the other three treatments. In fact, the ammonia trend for the other three treatments during the first 14 days of the newly established aquariums was not significantly different than the control treatment (not inoculated). There was no evidence that the addition of more commercial AOB blends to aquariums would reduce the amount of time needed to establish ammonia oxidation. These results demonstrate different points of view: (1) the AOB strains of the present invention represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 are different from the bacterial strains of the present invention represented by SEQ ID NO: 18 and SEQ ID NO: 19; (2) AOB contained in commercially available mixtures (reported to be Nitrosomonas europaea) are not effective in controlling ammonia during the startup of new seawater aquariums; and (3) that the AOB represented by SEQ ID NO: 18 and SEQ ID NO: 19 are effective in controlling and maintaining ammonia concentrations in newly established seawater aquariums.

EXAMPLE 15 Bacterial Additive Test IX The objective of this test was to compare the biomass material of the SB7 reactor (which contained the AOB strains of the present invention represented by SEQ ID NO: 18, SEQ ID NO: 19 and two similar strains to halophila) with aquariums that did not receive a bacterial inoculation. For this test, eight 37.854 liter (10 gallon) aquariums and eight penguin 170B (Marineland Aquarium Products) style energy filters hung on the back were sterilized, rinsed thoroughly and allowed to air dry. The next day, each tank was filled with 4,536 kilograms (10 lbs) of rinsed Tideline # 5 chopped coral and equipped with an esterified energy filter (PF 0170B) and rinsed with a carbon cartridge. Each tank was filled with 35 L of artificial seawater. The artificial seawater was added by adding SeaSalts INSTANT OCEAN (Aquarium Systems, Mentor, Ohio) to city water filtered with coal until the salinity was 30 ppt. The aquariums were filled with seawater and the filters were allowed to run overnight before the addition of the bacterial additives and the fish. The next morning the tanks were completely filled with ultrapure water to compensate for the evaporation and water mixtures taken. Then the four tanks were dosed with 150 ml of the bacterial mixture of the SB7 reactor. The other four tanks were not dosed with a bacterial mixture. The AOB SB7 reactor mixture consisted of AOB strains of the present invention represented by SEQ ID NO: 18, SEQ ID NO: 19 and two strains similar to N. halophila. Six clownfish (Amphiprion ocellaris) were added to each tank on the first day of the test and fed twice a day. The fish feed was a mixture of frozen brine shrimp and Spirulina fish flakes. On day 3 of the test, four additional clownfish were added to each aquarium. Water samples were collected and analyzed per day to measure pH, ammonia, nitrite and conductivity. On Monday, Wednesday and Friday, water was tested to measure nitrite and turbidity. The pH measurements were made with a model 225 pH / ion meter from Denver Instruments equipped with a pH combination electrode from Denver Instruments. A Tecator FIAstar 5010 analyzer was used to measure ammonia, nitrite and nitrate (as nitrogen) using the methods described in the Tecator application recommendations. Salinity was measured directly in each tank per day using a salinity, conductivity and temperature system fastened with the YSI model 30 hand. The turbidity data were determined with a turbidity meter DRT-100 (HF Scientific). The average ammonia concentrations for the SB7 treatment and control are presented in Figure 9. The SB7 treatment, which includes the AOB strains of the present invention represented by SEQ ID NO: 18, SEQ ID NO: 19 and two similar strains to N. halophila, oxidized ammonia considerably faster than the control. The average concentration of ammonia reached 0 mg / L on day 9 in the tanks that received the SB7 treatment, while 17 days passed in the control aquariums before the ammonia values reached the same level of 0 mg / L. In addition, the mean maximum ammonia concentration of the SB7 treatment (approximately 0.4 mg / L-N) was significantly lower than the control treatment (1.72 mg / L-N). The results demonstrate that the AOB strains of the present invention represented by SEQ ID NO: 18 and SEQ ID NO: 19 are effective in controlling ammonia concentrations in newly established aquariums.

EXAMPLE 16 X-Test of Bacterial Additive The objective of this test was to compare the biomass material of the SB7 reactor (which contained the AOB strains of the present invention represented by SEQ ID NO: 18, SEQ ID NO: 19 and two similar strains to halophila) and reactor B7 (which contained two strains of AOB similar to N. halophila) with aquariums that did not receive bacterial inoculation. For this test, ten 37.854 liter (10 gallon) aquariums and ten penguin 170B marine style energy filters (Marineland Aquarium Products) were sterilized, rinsed thoroughly and allowed to air dry. The next day, each tank was equipped with a sterilized energy filter (PF 0170B) and rinsed with a carbon cartridge. Each tank was filled with 19 L of artificial seawater. The artificial seawater was added by adding SeaSalts INSTANT OCEAN (Aquarium Systems, Mentor, Ohio) to city water filtered with coal until the salinity was 29 ppt. The aquariums were filled with seawater and the filters were allowed to run overnight before the addition of bacterial additives and ammonia.

The next morning the tanks were completely filled with ultrapure water to compensate for the evaporation and water mixtures taken. Each of the treatment and control tanks had four replicas. Four aquariums were dosed with 150 ml of the bacterial mixture of the SB7 reactor, four aquaria were dosed with 150 ml of the bacterial mixture of the B7 reactor and four aquariums were not dosed with any bacterial mixture. The AOB SB7 reactor mixture consisted of AOB strains of the present invention represented by SEQ ID NO: 18, SEQ ID NO: 19 and two strains similar to N. halophila. The AOB B7 reactor mixture consisted of two strains of AOB similar to N. halophila. Each day, 11.5 mg of ammonia-nitrogen was added to each aquarium. Water samples were collected and analyzed per day to measure pH, ammonia, nitrite and conductivity. On Monday, Wednesday and Friday, water was tested to measure nitrite and turbidity. The pH measurements were made with a model 225 pH / ion meter from Denver Instruments equipped with a pH combination electrode from Denver Instruments. A Tecator FIAstar 5010 analyzer was used to measure ammonia, nitrite and nitrate (as nitrogen) using the methods described in the Tecator application recommendations. Salinity was measured directly in each tank per day using a salinity, conductivity and temperature system fastened with the YSI model 30 hand. The turbidity data were determined with a turbidity meter DRT-100 (HF Scientific). The average ammonia concentrations for the two treatments and control are depicted in Figure 10. The ammonia values for the aquaria that received either the B7 reactor or the SB7 reactor oxidized the ammonia at almost the same rate; considerably faster than the control. The average concentration of ammonia reached 0 mg / L on day 3 for the tanks that received either the B7 or SB7 treatments, and the mean maximum ammonia concentration of the B7 and SB7 treatments (approximately 0.2 mg / LN) was significantly lower than the control treatment (2.0 mg / LN) (Fig. 9). The results demonstrate that the AOB strains of the present invention, represented by SEQ ID NO: 18 and SEQ ID NO: 19, are effective in controlling the concentration of ammonia in newly established aquariums. While the foregoing description refers to the particular embodiments of the present invention, it should be readily apparent to those skilled in the art that different modifications can be made without departing from the spirit thereof. The appended claims are intended to cover these modifications since they would be within the real spirit and scope of the invention. Thus, the modalities described at present will be considered in all aspects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims instead of the above description. All the changes that fall within the meaning of and a range of equivalency of the claims are intended to be encompassed therein. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims

REIVI DICATIONS Having described the invention as above, the content of the following claims is claimed as property: 1. A composition comprising an isolated bacterial strain that oxidizes ammonia to nitrite, characterized in that the bacterial strain comprises a nucleotide sequence established in a sequence selected from the group consisting of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20.
2. The composition in accordance with the claim 1, characterized in that the composition is in a form selected from the group consisting of liquid, frozen, dried by freezing and pulverized. 3. The composition according to claim 1, characterized in that the composition is included in a polymer. 4. The composition according to claim 3, characterized in that the polymer is selected from the group consisting of acrylamide, alginate, carrageenan and combinations thereof. 5. A composition comprising a concentrated bacterial strain that oxidizes ammonia to nitrite, characterized in that the 16S rDNA of the bacterial strain has a nucleotide sequence selected from the group consisting of: a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 18, a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 19 and a nucleotide sequence having at least 96 % identity over the total length thereof to SEQ ID NO. 20. The composition according to claim 5, characterized in that the bacterial strain has a 16S rDNA sequence which is identical to a sequence selected from the group consisting of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 18. NO: 20. The composition according to claim 5, characterized in that it also comprises a microorganism selected from the group consisting of microorganisms that oxidize ammonia, microorganisms that oxidize nitrite, microorganisms that reduce nitrate, heterotrophic microorganisms and combinations of the same. 8. An isolated nucleic acid, characterized in that it is selected from the group consisting of: a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 18, a nucleotide sequence which has at least 96% identity over the total length thereof at SEQ ID NO: 19 and a nucleotide sequence having at least 96% identity over the total length thereof at SEQ ID NO. twenty. 9. The isolated nucleic acid according to claim 8, characterized in that it is identical to a sequence selected from the group consisting of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20. 10. An oligonucleotide probe, characterized in that it comprises the nucleotide sequence as set forth in SEQ ID NO: 21. 11. An oligonucleotide probe having at least 96% identity over the total length thereof to the nucleotide sequence set forth in SEQ ID NO: 21, characterized in that hybrid to the nucleic acid of the bacteria having 16S rDNA having a nucleotide sequence having at least 96% identity over the total length thereof, to a sequence selected from the group consisting of SEQ ID NO . 18, SEQ ID NO: 19 and SEQ ID NO: 20. 12. A polymerase chain reaction (PCR) primer, characterized in that it is selected from the group consisting of SEQ ID NO: 22 and SEQ ID NO: 23 13. A polymerase chain reaction (PCR) primer having at least 96% identity over the total length thereof to a nucleotide sequence selected from the group consisting of SEQ ID NO: 22 and SEQ ID NO. NO: 23, characterized in that the PCR primer hybridized to the nucleic acid of the bacteria having 16S rDNA having a nucleotide sequence having at least 96% identity over the total length thereof, to a sequence selected from the group consisting of SEQ ID NO. 18, SEQ ID NO: 19 and SEQ ID NO: 20. 14. A composition comprising at least two bacterial strains that oxidize ammonia to nitrite, characterized in that each of at least two bacterial strains has the 16S rDNA that includes a nucleotide sequence independently selected from the group consisting of: a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 18, a nucleotide sequence having at least 96% identity about the total length thereof to SEQ ID NO: 19 and a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO. 20. The composition according to claim 14, characterized in that it comprises a bacterial strain with a 16S rDNA that includes a sequence selected as set forth in SEQ ID NO: 18, a bacterial strain with a 16S rDNA that includes a sequence nucleotide as set forth in SEQ ID NO: 19 and a bacterial strain with a 16S rDNA that includes a nucleotide sequence as set forth in SEQ ID NO: 20. 16. A method for reducing or preventing the accumulation of ammonia in a medium , characterized in that it comprises: providing a bacterial strain that oxidizes ammonia to nitrite, wherein the bacterial strain comprises a nucleotide sequence selected from the group consisting of: a nucleotide sequence having more than 96% identity over the total length thereof to SEQ ID NO: 18, a nucleotide sequence having more than 96% identity over the total length thereof to SEQ ID NO: 19 and a nucleotide sequence which It has more than 96% identity over the total length of it to SEC ID NO. twenty; and introducing in the medium an amount of the bacterial strain sufficient to reduce or prevent the accumulation of ammonia in the medium. The method according to claim 16, characterized in that the ammonia is reduced to at least 30% compared to a level of ammonia that would exist in the absence of the bacterial strain. 18. The method according to claim 16, characterized in that the medium is an aquarium. 19. The method according to claim 18, characterized in that the aquarium is a freshwater aquarium. 20. The method according to claim 18, characterized in that the aquarium is an aquarium of seawater. 21. The method according to claim 16, characterized in that the medium comprises waste water. 22. The method according to claim 16, characterized in that an amount of the bacterial strain which also comprises placing the bacterial strain in the medium in a rotating biological contactor is introduced into the medium. '2.
3. The method according to claim 16, characterized in that an amount of the bacterial strain which also comprises placing the bacterial strain in the medium in a biofilter is introduced into the medium. 24. A method for reducing or preventing the accumulation of ammonia in a medium, characterized in that it comprises: providing a bacterial strain that oxidizes ammonia to nitrite, wherein the bacterial strain comprises a 16S rDNA nucleotide sequence established in a sequence selected from the group which consists of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20; and introducing in the medium an amount of the bacterial strain sufficient to reduce or prevent the accumulation of ammonia in the medium. 25. A bioremediation process that reduces or prevents the accumulation of ammonia in a medium, characterized in that it comprises: providing a bacterial strain that oxidizes ammonia to nitrite, wherein the bacterial strain comprises a nucleotide sequence selected from the group consisting of: nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 18, a nucleotide sequence having at least 96% identity "over the total length thereof to SEQ ID NO. NO: 19 and a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 20, and introducing into the medium an amount of the bacterial strain sufficient to remedy the medium. A method for reducing or preventing the accumulation of ammonia in a medium, characterized in that it comprises: providing a composition comprising at least two bacterial strains that oxidize ammonia to nitrite, where e each of the two bacterial strains has a 16S rDNA that includes a nucleotide sequence independently selected from the group consisting of: a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 18, a nucleotide sequence having at least 96% identity over the total length thereof to SEQ ID NO: 19 and a nucleotide sequence having at least 96% identity over the total length thereof at SEC ID NO. twenty; and introducing in the medium a quantity of the composition, sufficient to reduce or prevent the accumulation of ammonia in the medium. 27. The method according to claim 26, characterized in that the composition comprises a bacterial strain with a 16S rDNA that includes a nucleotide sequence as set forth in SEQ ID NO: 18, a bacterial strain with a 16S rDNA that includes a sequence nucleotide as set forth in SEQ ID NO: 19 and a bacterial strain with a 16S rDNA that includes a nucleotide sequence as set forth in SEQ ID NO: 20. 28. A method for detecting and determining the amount of bacteria that oxidize the ammonia to nitrite in a medium, characterized in that the 16S rDNA of the bacteria includes a nucleotide sequence selected from the group consisting of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 comprising: providing a probe detectably labeled which includes a nucleotide sequence as set forth in SEQ ID NO: 21; Isolate the total DNA from the medium; exposing the isolated total DNA to the probe detectably labeled under conditions under which the probe hybridizes only the nucleic acid of the bacteria having the 16S rDNA including the nucleotide sequence; and detect and measure the amount of hybridized probe, wherein the presence of the hybridized probe is indicative of the presence of bacteria that oxidize ammonia to nitrite and the amount of the hybridized probe is indicative of the amount of bacteria that oxidize ammonia to nitrite in the medium. 29. The method according to claim 28, characterized in that the medium is selected from the group consisting of aquarium water, fresh water, salt water and waste water. 30. The method according to claim 28, characterized in that the medium includes a material selected from the group consisting of aquarium gravel, filter sponges, filter cloth and plastic filter media. 31. The method according to claim 30, characterized in that the total DNA is isolated from the material. 32. The method according to claim 28, characterized in that it provides a detectably labeled probe which further comprises including the probe detectably labeled in a DNA portion. 33. The method according to claim 28, characterized in that the method for detecting and determining the amount of bacteria that oxidize ammonia to nitrite in a medium is an automated process. 34. The method according to claim 33, characterized in that the automated process is selected from the group consisting of DNA microarray, protein microarray, biosensors, biosonde, capillary electrophoresis and real-time PCR. 35. A method for detecting and determining the amount of bacteria that oxidize ammonia to nitrite in a medium, characterized in that the 16S rDNA of the bacteria includes a nucleotide sequence selected from the group consisting of: a nucleotide sequence having at least 96 % identity over the total length thereof to SEQ ID NO: 18, at least 96% identity over the total length thereof to SEQ ID NO: 19 and at least 96% identity over the length total thereof to SEQ ID NO: 20 comprising: providing a detectably labeled probe that includes at least 96% identity over the total length thereof to a nucleotide sequence as set forth in SEQ ID NO: 21; Isolate the total DNA from the medium; exposing the isolated total DNA to the probe detectably labeled under conditions under which the probe hybridizes only the nucleic acid of the bacteria having the 16S rDNA including the nucleotide sequence; and detecting and measuring the amount of hybridized probe, wherein the presence of the hybridized probe is indicative of the presence of bacteria that oxidize ammonia to nitrite and the amount of the hybridized probe is indicative of the amount of bacteria that oxidize ammonia to nitrite in the middle. 36. The method according to claim 35, characterized in that the medium is selected from the group consisting of aquarium water, fresh water, salt water and waste water. 37. The method according to claim 35, characterized in that the medium includes a material selected from the group consisting of aquarium gravel, filter sponges, filter silk and plastic filter media. 38. The method according to claim 37, characterized in that the total DNA is isolated from the material. 39. The method according to claim 35, characterized in that it provides a detectably labeled probe comprising comprising the probe detectably labeled in a portion of DNA. 40. The method according to claim 35, characterized in that the method for detecting and determining the amount of bacteria that oxidize ammonia to nitrite in a medium is an automated process. 41. The method according to claim 40, characterized in that the automated process is selected from the group consisting of DNA microarray, protein microarray, biosensors, biosonde, capillary electrophoresis and real-time PCR. 42. A portion of DNA, characterized in that it comprises: a solid substrate; and a probe that includes a nucleotide sequence as set forth in SEQ ID NO: 21 configured on the solid substrate. 43. The DNA portion according to claim 42, characterized in that the probe is bound to the solid substrate by a covalent bond.