Scalindua Wagneri
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Candidatus In prokaryote nomenclature, ''Candidatus'' (Latin for candidate of Roman office) is used to name prokaryotic phyla that are well characterized but yet-uncultured. Contemporary sequencing approaches, such as 16S sequencing or metagenomics, provide m ...
'' Scalindua wagneri is a
Gram-negative Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. They are characterized by their cell envelopes, which are composed of a thin peptidoglycan cell wall ...
coccoid-shaped bacterium that was first isolated from a wastewater treatment plant. This bacterium is an obligate
anaerobic Anaerobic means "living, active, occurring, or existing in the absence of free oxygen", as opposed to aerobic which means "living, active, or occurring only in the presence of oxygen." Anaerobic may also refer to: * Anaerobic adhesive, a bonding a ...
chemolithotroph Lithotrophs are a diverse group of organisms using an inorganic substrate (usually of mineral origin) to obtain reducing equivalents for use in biosynthesis (e.g., carbon dioxide fixation) or energy conservation (i.e., ATP production) via aerobi ...
that undergoes anaerobic ammonium oxidation (
anammox Anammox, an abbreviation for anaerobic ammonium oxidation, is a globally important microbial process of the nitrogen cycle that takes place in many natural environments. The bacteria mediating this process were identified in 1999, and were a grea ...
). It can be used in the wastewater treatment industry in nitrogen reactors to remove nitrogenous wastes from wastewater without contributing to fixed nitrogen loss and
greenhouse gas emission Greenhouse gas emissions from human activities strengthen the greenhouse effect, contributing to climate change. Most is carbon dioxide from burning fossil fuels: coal, oil, and natural gas. The largest emitters include coal in China and ...
.


Characterization

''Candidatus'' Scalindua wagneri is a coccoid-shaped bacterium with a diameter of 1 μm. Like other
Planctomycetota The Planctomycetota are a phylum of widely distributed bacteria, occurring in both aquatic and terrestrial habitats. They play a considerable role in global carbon and nitrogen cycles, with many species of this phylum capable of anaerobic ammoniu ...
, ''S. wagneri'' is
Gram-negative Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. They are characterized by their cell envelopes, which are composed of a thin peptidoglycan cell wall ...
and does not have
peptidoglycan Peptidoglycan or murein is a unique large macromolecule, a polysaccharide, consisting of sugars and amino acids that forms a mesh-like peptidoglycan layer outside the plasma membrane, the rigid cell wall (murein sacculus) characteristic of most ...
in its
cell wall A cell wall is a structural layer surrounding some types of cells, just outside the cell membrane. It can be tough, flexible, and sometimes rigid. It provides the cell with both structural support and protection, and also acts as a filtering mech ...
. In addition, the bacterium contains two inner membranes instead of having one inner membrane and one outer membrane that surrounds the cell wall. Some of the near neighbors are other species within the new ''Scalindua'' genus, such as "''Candidatus'' S. sorokinii" and "''Candidatus'' S. brodae". Other neighbors include "''Candidatus'' Kuenenia stuttgartiensis" and "''Candidatus''
Brocadia anammoxidans "''Candidatus'' Brocadia" is a ''candidatus'' genus of bacteria. Many of the species in this genus are capable of anammox. Phylogeny Phylogeny based on GTDB 07-RS207 by Genome Taxonomy Database The Genome Taxonomy Database (GTDB) is an onl ...
". S. wagneri and its genus share only about 85% similarity with other members in its evolutionary line, which suggests that it is distantly related to other anaerobic ammonium oxidizing (anammox) bacteria.


Discovery

Markus Schmid from the Strous lab first discovered S. wagneri in a landfill
leachate A leachate is any liquid that, in the course of passing through matter, extracts soluble or suspended solids, or any other component of the material through which it has passed. Leachate is a widely used term in the environmental sciences wher ...
treatment plant located in
Pitsea Pitsea is a small town and former civil parish, now in the unparished area of Basildon, in south Essex, England. It comprises five sub-districts: Eversley, Northlands Park Neighbourhood (previously known as Felmores), Chalvedon, Pitsea Mount and B ...
, UK on August 1, 2001. These bacteria doubled in number about every three weeks in laboratory conditions, which made them very difficult to isolate. Therefore, the researchers used 16S
rRNA Ribosomal ribonucleic acid (rRNA) is a type of non-coding RNA which is the primary component of ribosomes, essential to all cells. rRNA is a ribozyme which carries out protein synthesis in ribosomes. Ribosomal RNA is transcribed from ribosoma ...
(
ribosomal RNA Ribosomal ribonucleic acid (rRNA) is a type of non-coding RNA which is the primary component of ribosomes, essential to all cells. rRNA is a ribozyme which carries out protein synthesis in ribosomes. Ribosomal RNA is transcribed from ribosomal ...
) gene analysis on the
biofilm A biofilm comprises any syntrophic consortium of microorganisms in which cells stick to each other and often also to a surface. These adherent cells become embedded within a slimy extracellular matrix that is composed of extracellular ...
of wastewater samples to detect the presence of these bacteria. They amplified and isolated the 16S rRNA gene from the biofilm using PCR and
gel electrophoresis Gel electrophoresis is a method for separation and analysis of biomacromolecules ( DNA, RNA, proteins, etc.) and their fragments, based on their size and charge. It is used in clinical chemistry to separate proteins by charge or size (IEF ...
. Then, they
cloned Cloning is the process of producing individual organisms with identical or virtually identical DNA, either by natural or artificial means. In nature, some organisms produce clones through asexual reproduction. In the field of biotechnology, c ...
the DNA into TOPO vectors. Once the researchers sequenced the DNA, they aligned the 16S rRNA gene sequences to a
genome In the fields of molecular biology and genetics, a genome is all the genetic information of an organism. It consists of nucleotide sequences of DNA (or RNA in RNA viruses). The nuclear genome includes protein-coding genes and non-coding ge ...
database and found that the sequences are related to the anammox bacteria. One of the sequences showed a 93% similarity to ''Candidatus'' Scalindua sorokinii, which suggests that this sequence belonged to a new species within the genus Scalindua and the researchers named it ''Candidatus'' Scalindua wagneri after Michael Wagner, a microbial
ecologist Ecology () is the study of the relationships between living organisms, including humans, and their physical environment. Ecology considers organisms at the individual, population, community, ecosystem, and biosphere level. Ecology overlaps wi ...
.


Metabolism

S. wagneri is an obligate
anaerobic Anaerobic means "living, active, occurring, or existing in the absence of free oxygen", as opposed to aerobic which means "living, active, or occurring only in the presence of oxygen." Anaerobic may also refer to: * Anaerobic adhesive, a bonding a ...
chemolithoautotroph A lithoautotroph is an organism which derives energy from reactions of reduced compounds of mineral (inorganic) origin. Two types of lithoautotrophs are distinguished by their energy source; photolithoautotrophs derive their energy from light while ...
and undergoes anaerobic ammonium oxidation (anammox) in the intracytoplasmic compartment called an
anammox Anammox, an abbreviation for anaerobic ammonium oxidation, is a globally important microbial process of the nitrogen cycle that takes place in many natural environments. The bacteria mediating this process were identified in 1999, and were a grea ...
osome. During the anammox process,
ammonium The ammonium cation is a positively-charged polyatomic ion with the chemical formula or . It is formed by the protonation of ammonia (). Ammonium is also a general name for positively charged or protonated substituted amines and quaternary a ...
is oxidized using
nitrite The nitrite polyatomic ion, ion has the chemical formula . Nitrite (mostly sodium nitrite) is widely used throughout chemical and pharmaceutical industries. The nitrite anion is a pervasive intermediate in the nitrogen cycle in nature. The name ...
as an
electron acceptor An electron acceptor is a chemical entity that accepts electrons transferred to it from another compound. It is an oxidizing agent that, by virtue of its accepting electrons, is itself reduced in the process. Electron acceptors are sometimes mista ...
and forms dinitrogen gas as a product. It is proposed that this mechanism occurs through the production of a
hydrazine Hydrazine is an inorganic compound with the chemical formula . It is a simple pnictogen hydride, and is a colourless flammable liquid with an ammonia-like odour. Hydrazine is highly toxic unless handled in solution as, for example, hydrazine ...
intermediate using
hydroxylamine Hydroxylamine is an inorganic compound with the formula . The material is a white crystalline, hygroscopic compound.Greenwood and Earnshaw. ''Chemistry of the Elements.'' 2nd Edition. Reed Educational and Professional Publishing Ltd. pp. 431–43 ...
, which is derived from nitrite. In addition, S. wagneri uses nitrite as an
electron donor In chemistry, an electron donor is a chemical entity that donates electrons to another compound. It is a reducing agent that, by virtue of its donating electrons, is itself oxidized in the process. Typical reducing agents undergo permanent chem ...
to fix
carbon dioxide Carbon dioxide (chemical formula ) is a chemical compound made up of molecules that each have one carbon atom covalently double bonded to two oxygen atoms. It is found in the gas state at room temperature. In the air, carbon dioxide is transpar ...
and forms
nitrate Nitrate is a polyatomic ion A polyatomic ion, also known as a molecular ion, is a covalent bonded set of two or more atoms, or of a metal complex, that can be considered to behave as a single unit and that has a net charge that is not zer ...
as a byproduct. To the test the
metabolic Metabolism (, from el, μεταβολή ''metabolē'', "change") is the set of life-sustaining chemical reactions in organisms. The three main functions of metabolism are: the conversion of the energy in food to energy available to run cell ...
properties of S. wagneri, Nakajima ''et al''. performed anammox activity tests using nitrogen compounds labeled with the 15 N isotopes and measured 28N2, 29N2, and 30N2 concentrations after 15 days. The researchers found that the concentrations of the 28N2 and 29N2 gases increased significantly. These results suggest that ammonia and nitrite is used in equal amounts to make 29N2, and
denitrification Denitrification is a microbially facilitated process where nitrate (NO3−) is reduced and ultimately produces molecular nitrogen (N2) through a series of intermediate gaseous nitrogen oxide products. Facultative anaerobic bacteria perform denitr ...
concurrently occurs with anammox metabolism.


Genome

Currently, genomic information about S. wagneri is very limited. Current genome sequences were collected from DNA isolated from the bacteria growing in a marine anammox bacteria (MAB) reactor. Then, the 16S rRNA genes on the DNA were amplified using a specific
oligonucleotide Oligonucleotides are short DNA or RNA molecules, oligomers, that have a wide range of applications in genetic testing, research, and forensics. Commonly made in the laboratory by solid-phase chemical synthesis, these small bits of nucleic acids c ...
primer Primer may refer to: Arts, entertainment, and media Films * ''Primer'' (film), a 2004 feature film written and directed by Shane Carruth * ''Primer'' (video), a documentary about the funk band Living Colour Literature * Primer (textbook), a t ...
for ''Planctomycetales'', separated using gel electrophoresis, and sequenced using a CEQ 2000 DNA Sequencer. Analysis of the 16S rRNA gene sequences was performed using the GENETYX program, and the alignments and phylogenetic trees were made using
BLAST Blast or The Blast may refer to: * Explosion, a rapid increase in volume and release of energy in an extreme manner *Detonation, an exothermic front accelerating through a medium that eventually drives a shock front Film * ''Blast'' (1997 film) ...
,
CLUSTALW Clustal is a series of widely used computer programs used in bioinformatics for multiple sequence alignment. There have been many versions of Clustal over the development of the algorithm that are listed below. The analysis of each tool and its ...
and
neighbor joining In bioinformatics, neighbor joining is a bottom-up (agglomerative) clustering method for the creation of phylogenetic trees, created by Naruya Saitou and Masatoshi Nei in 1987. Usually based on DNA or protein sequence data, the algorithm require ...
, respectively. To have a better understanding of the
genome In the fields of molecular biology and genetics, a genome is all the genetic information of an organism. It consists of nucleotide sequences of DNA (or RNA in RNA viruses). The nuclear genome includes protein-coding genes and non-coding ge ...
, S. wagneri can be compared to one of its better-known relatives. For example, ''Candidatus'' Scalindua profunda has a genome length of 5.14 million base pairs with a
GC content In molecular biology and genetics, GC-content (or guanine-cytosine content) is the percentage of nitrogenous bases in a DNA or RNA molecule that are either guanine (G) or cytosine (C). This measure indicates the proportion of G and C bases out o ...
of 39.1%. There is no genomic information about the length or % GC content for S. wagneri. However, there are hundreds of 476 base pair partial sequences for its 16S rRNA gene. Using
fluorescent in situ hybridization Fluorescence ''in situ'' hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only particular parts of a nucleic acid sequence with a high degree of sequence complementarity. It was developed ...
(FISH) analysis, a technique used to detect specific DNA sequences on
chromosomes A chromosome is a long DNA molecule with part or all of the genetic material of an organism. In most chromosomes the very long thin DNA fibers are coated with packaging proteins; in eukaryotic cells the most important of these proteins are ...
, researchers were not able to detect hybridization between the chromosome of S. wagneri and the putative anammox DNA probe. This suggests that S. wagneri is not very similar to the known anammox bacteria, so the researchers categorized the bacterium into its own genus.


Ecology

Although researchers are unable to isolate pure
cultures Culture () is an umbrella term which encompasses the social behavior, institutions, and norms found in human societies, as well as the knowledge, beliefs, arts, laws, customs, capabilities, and habits of the individuals in these groups.Tyl ...
of S. wagneri, it is believed to encompass a broad
niche Niche may refer to: Science *Developmental niche, a concept for understanding the cultural context of child development *Ecological niche, a term describing the relational position of an organism's species *Niche differentiation, in ecology, the ...
. Using 16S rRNA gene analysis, Schmid first found evidence of the bacteria in wastewater treatment plants. Other researchers also found 16S rRNA gene evidence in a petroleum reservoir held at a temperature range between 55 °C and 75 °C in addition to
freshwater Fresh water or freshwater is any naturally occurring liquid or frozen water containing low concentrations of dissolved salts and other total dissolved solids. Although the term specifically excludes seawater and brackish water, it does include ...
and
marine Marine is an adjective meaning of or pertaining to the sea or ocean. Marine or marines may refer to: Ocean * Maritime (disambiguation) * Marine art * Marine biology * Marine debris * Marine habitats * Marine life * Marine pollution Military * ...
ecosystems An ecosystem (or ecological system) consists of all the organisms and the physical environment with which they interact. These biotic and abiotic components are linked together through nutrient cycles and energy flows. Energy enters the syste ...
, such as
estuaries An estuary is a partially enclosed coastal body of brackish water with one or more rivers or streams flowing into it, and with a free connection to the open sea. Estuaries form a transition zone between river environments and maritime environment ...
.


Importance and useful applications

S. wagneri allows wastewater treatment plants to reduce operation costs while reducing the adverse effects of
nitrification ''Nitrification'' is the biological oxidation of ammonia to nitrite followed by the oxidation of the nitrite to nitrate occurring through separate organisms or direct ammonia oxidation to nitrate in comammox bacteria. The transformation of amm ...
and
denitrification Denitrification is a microbially facilitated process where nitrate (NO3−) is reduced and ultimately produces molecular nitrogen (N2) through a series of intermediate gaseous nitrogen oxide products. Facultative anaerobic bacteria perform denitr ...
on the environment. These bacteria contribute to the development of new technologies for wastewater management by aiding in the efficient removal of nitrogenous compounds in wastewater. Usually, nitrogen reactors use both nitrification and denitrification to remove nitrogenous wastes. These processes have high operation costs due to the continuous maintenance of
aerobic Aerobic means "requiring air," in which "air" usually means oxygen. Aerobic may also refer to * Aerobic exercise, prolonged exercise of moderate intensity * Aerobics, a form of aerobic exercise * Aerobic respiration, the aerobic process of cellu ...
conditions in the reactor. Denitrification also produces
nitrous oxide Nitrous oxide (dinitrogen oxide or dinitrogen monoxide), commonly known as laughing gas, nitrous, or nos, is a chemical compound, an oxide of nitrogen with the formula . At room temperature, it is a colourless non-flammable gas, and has a ...
(N2O), which is a greenhouse gas that is detrimental to the environment. Production of N2O contributes to the loss of fixed nitrogen, which regulates the biological
productivity Productivity is the efficiency of production of goods or services expressed by some measure. Measurements of productivity are often expressed as a ratio of an aggregate output to a single input or an aggregate input used in a production proces ...
of
ecosystems An ecosystem (or ecological system) consists of all the organisms and the physical environment with which they interact. These biotic and abiotic components are linked together through nutrient cycles and energy flows. Energy enters the syste ...
. By inoculating wastewater reactors with the
anaerobic Anaerobic means "living, active, occurring, or existing in the absence of free oxygen", as opposed to aerobic which means "living, active, or occurring only in the presence of oxygen." Anaerobic may also refer to: * Anaerobic adhesive, a bonding a ...
S. wagneri, operation costs can be reduced by about ninety percent without the production of greenhouse gases. This allows for better wastewater management in a more cost-efficient manner without contributing to
climate change In common usage, climate change describes global warming—the ongoing increase in global average temperature—and its effects on Earth's climate system. Climate change in a broader sense also includes previous long-term changes to E ...
.


References

{{Taxonbar, from=Q7429778 Environmental microbiology Planctomycetota Bacteria described in 2003 Candidatus taxa