Macromonas Bipunctata
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''Macromonas'' ''bipunctata'' 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 ...
, colorless, and
heterotroph A heterotroph (; ) is an organism that cannot produce its own food, instead taking nutrition from other sources of organic carbon, mainly plant or animal matter. In the food chain, heterotrophs are primary, secondary and tertiary consumers, but ...
ic sulfur bacterium of the
genus Genus ( plural genera ) is a taxonomic rank used in the biological classification of extant taxon, living and fossil organisms as well as Virus classification#ICTV classification, viruses. In the hierarchy of biological classification, genus com ...
'' Macromonas''. It is commonly found in sewage aeration tanks and caves where
moonmilk Moonmilk (sometimes called mondmilch, also known as montmilch or as cave milk) is a white, creamy substance found inside limestone, dolomite, and possibly other types of caves. It is a precipitate from limestone comprising aggregates of fine cry ...
has formed. In the 1920s, researcher Gicklhorn first discovered this organism under the name ''
Pseudomonas ''Pseudomonas'' is a genus of Gram-negative, Gammaproteobacteria, belonging to the family Pseudomonadaceae and containing 191 described species. The members of the genus demonstrate a great deal of metabolic diversity and consequently are able ...
'' ''bipunctata''. After further study and culturing by Utermöhl and Koppe, in 1923, it was later renamed '' Macromonas'' ''bipunctata''. This organism is thought to be non-
pathogenic In biology, a pathogen ( el, πάθος, "suffering", "passion" and , "producer of") in the oldest and broadest sense, is any organism or agent that can produce disease. A pathogen may also be referred to as an infectious agent, or simply a germ ...
species. In fact, the moonmilk produced was referenced as a remedy for infections in the Middle Ages.


Background


History

In the Middle Ages, "
moonmilk Moonmilk (sometimes called mondmilch, also known as montmilch or as cave milk) is a white, creamy substance found inside limestone, dolomite, and possibly other types of caves. It is a precipitate from limestone comprising aggregates of fine cry ...
" was used as a medicine. People often used it to cure infections and accelerate the healing process. Moonmilk is more than simply '' Macromonas bipunctata''. It also contains populations of
cyanobacteria Cyanobacteria (), also known as Cyanophyta, are a phylum of gram-negative bacteria that obtain energy via photosynthesis. The name ''cyanobacteria'' refers to their color (), which similarly forms the basis of cyanobacteria's common name, blu ...
,
fungi A fungus ( : fungi or funguses) is any member of the group of eukaryotic organisms that includes microorganisms such as yeasts and molds, as well as the more familiar mushrooms. These organisms are classified as a kingdom, separately from ...
,
green algae The green algae (singular: green alga) are a group consisting of the Prasinodermophyta and its unnamed sister which contains the Chlorophyta and Charophyta/Streptophyta. The land plants (Embryophytes) have emerged deep in the Charophyte alga as ...
, and
actinomycetes The Actinomycetales is an order of Actinomycetota. A member of the order is often called an actinomycete. Actinomycetales are generally gram-positive and anaerobic and have mycelia in a filamentous and branching growth pattern. Some actinomycete ...
, which are the main producers of
antibiotics An antibiotic is a type of antimicrobial substance active against bacteria. It is the most important type of antibacterial agent for fighting bacterial infections, and antibiotic medications are widely used in the treatment and prevention o ...
. This could explain why it was effective as a potential agent for healing.


Etymology

''M. bipunctata'' was first isolated by Gicklhorn in the slime of a large basin Gratz in a botanical garden in 1924. Gicklhorn treated this species as a colorless sulfur bacteria and called it ''
Pseudomonas ''Pseudomonas'' is a genus of Gram-negative, Gammaproteobacteria, belonging to the family Pseudomonadaceae and containing 191 described species. The members of the genus demonstrate a great deal of metabolic diversity and consequently are able ...
'' ''bipunctata''. The Greek root "monad/monas" was commonly used for microbiology to indicate a
unicellular A unicellular organism, also known as a single-celled organism, is an organism that consists of a single cell, unlike a multicellular organism that consists of multiple cells. Organisms fall into two general categories: prokaryotic organisms and ...
or single unit organism(s)/bacterium in the 1920s. Furthermore, ''bipunctata'' can be separated into the Latin roots "bi", meaning two, and "punctata" , meaning spotted, as seen in cultured ''M''. ''bipunctata''. Years later, Dubinina, Grabovich, and La Rivière isolated this species from the precipitates of sewage aeration tanks called the white mat. Upon more research of this organism, it was renamed '' Macromonas'' ''bipunctata''. "Macro" is the Greek term for large, as the cell itself is on average larger than most bacteria. Additionally, this species can also be found in many caves where
moonmilk Moonmilk (sometimes called mondmilch, also known as montmilch or as cave milk) is a white, creamy substance found inside limestone, dolomite, and possibly other types of caves. It is a precipitate from limestone comprising aggregates of fine cry ...
is present.


Microbiology

'' Macromonas'' ''bipunctata'' 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 ...
,
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 ...
, irregular/pear shaped,
heterotroph A heterotroph (; ) is an organism that cannot produce its own food, instead taking nutrition from other sources of organic carbon, mainly plant or animal matter. In the food chain, heterotrophs are primary, secondary and tertiary consumers, but ...
ic sulfur bacterium. M. bipunctata has a very large cell area at 9 µm x 20 µm . Its motility consists of
flagella A flagellum (; ) is a hairlike appendage that protrudes from certain plant and animal sperm cells, and from a wide range of microorganisms to provide motility. Many protists with flagella are termed as flagellates. A microorganism may have f ...
20–40 µm long that moves around using a structural beam of polar flagella located at one end of its body.


Phylogeny and taxonomy

The closest species to ''Macromonas'' ''bipunctata'' within the class
Betaproteobacteria Betaproteobacteria are a class of Gram-negative bacteria, and one of the eight classes of the phylum Pseudomonadota (synonym Proteobacteria). The ''Betaproteobacteria'' are a class comprising over 75 genera and 400 species of bacteria. Togeth ...
are ''Malikia'' ''granosa'' and ''Malikia'' ''spinosa'' based on
16S rRNA 16S rRNA may refer to: * 16S ribosomal RNA 16 S ribosomal RNA (or 16 S rRNA) is the RNA component of the 30S subunit of a prokaryotic ribosome ( SSU rRNA). It binds to the Shine-Dalgarno sequence and provides most of the SSU structure. The g ...
gene as shown in many previous studies. '' Malikia'' nests within the family
Comamonadaceae The Comamonadaceae are a family of the Betaproteobacteria.Willems A., J. De Ley, M. Gillis, and K. Kersters. ''Comamonadaceae, a New Family Encompassing the Acidovorans rRNA Complex, Including Variovorax paradoxus gen. nov.,comb. nov. for Alcali ...
in the phylum
Pseudomonadota Pseudomonadota (synonym Proteobacteria) is a major phylum of Gram-negative bacteria. The renaming of phyla in 2021 remains controversial among microbiologists, many of whom continue to use the earlier names of long standing in the literature. The ...
and is also
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 ...
. ''Malikia granosa'' has a 96.5% similarity to ''M. bipunctata'', whereas ''Hydrogenophaga flava'' has a 95.61% similarity in its 16S rRNA gene.


Culturing

Most of the culturing procedures model Dubinina and Grabovich's 1984 article on ''M. bipunctata'': it includes sodium acetate (1 g/L), calcium chloride (0.1 g/L), casein hydrolysate (0.1g/L), yeast extract (0.1g/L), and agar (1g/L) along with a vitamin supplement, trace elements, and FeS as a sulfide source. ''M. bipunctata'' was cultured on an
agar Agar ( or ), or agar-agar, is a jelly-like substance consisting of polysaccharides obtained from the cell walls of some species of red algae, primarily from ogonori (''Gracilaria'') and "tengusa" (''Gelidiaceae''). As found in nature, agar is ...
plate for 2–3 days at 28 °C (
mesophile A mesophile is an organism that grows best in moderate temperature, neither too hot nor too cold, with an optimum growth range from . The optimum growth temperature for these organisms is 37°C. The term is mainly applied to microorganisms. Organi ...
as optimum for cultivation set at around 28 degrees) before several species of '' Macromonas'' ''bipunctata'' appeared. The optimal pH level for growing is around 7.2–7.4. The colonies that form produce a white film on the surface of the plate along with flat, finegrained colonies of 1–4 mm diameter. ''M. bipunctata'' has a cell area at 9 µm x 20 µm . This species is also pear-shaped, gram-negative and catalase positive.


Genomics

Many of the studies using ''M. bipunctata'' still rely heavily on its morphological characteristics. However, it has been used as a phylogenetic comparison frequently so its 16s rRNA is catalogued: it is 1461 bp. The same study shows that the genome contains 67.6% GC content.


Metabolism

'' Macromonas'' ''bipunctata'' has been cultured in many studies that show H2O2 is formed in different biochemical reactions: not only in the process of
respiration Respiration may refer to: Biology * Cellular respiration, the process in which nutrients are converted into useful energy in a cell ** Anaerobic respiration, cellular respiration without oxygen ** Maintenance respiration, the amount of cellul ...
with the participation of
enzyme Enzymes () are proteins that act as biological catalysts by accelerating chemical reactions. The molecules upon which enzymes may act are called substrates, and the enzyme converts the substrates into different molecules known as products. A ...
s of the
electron transport chain An electron transport chain (ETC) is a series of protein complexes and other molecules that transfer electrons from electron donors to electron acceptors via redox reactions (both reduction and oxidation occurring simultaneously) and couples th ...
, but also in the course of utilization of intracellular
oxalate Oxalate (IUPAC: ethanedioate) is an anion with the formula C2O42−. This dianion is colorless. It occurs naturally, including in some foods. It forms a variety of salts, for example sodium oxalate (Na2C2O4), and several esters such as dimethyl o ...
inclusions in the
cytoplasm In cell biology, the cytoplasm is all of the material within a eukaryotic cell, enclosed by the cell membrane, except for the cell nucleus. The material inside the nucleus and contained within the nuclear membrane is termed the nucleoplasm. The ...
.
Oxidation Redox (reduction–oxidation, , ) is a type of chemical reaction in which the oxidation states of substrate change. Oxidation is the loss of electrons or an increase in the oxidation state, while reduction is the gain of electrons or a d ...
of
oxalate Oxalate (IUPAC: ethanedioate) is an anion with the formula C2O42−. This dianion is colorless. It occurs naturally, including in some foods. It forms a variety of salts, for example sodium oxalate (Na2C2O4), and several esters such as dimethyl o ...
inclusions by
oxalate Oxalate (IUPAC: ethanedioate) is an anion with the formula C2O42−. This dianion is colorless. It occurs naturally, including in some foods. It forms a variety of salts, for example sodium oxalate (Na2C2O4), and several esters such as dimethyl o ...
oxidase In biochemistry, an oxidase is an enzyme that catalyzes oxidation-reduction reactions, especially one involving dioxygen (O2) as the electron acceptor. In reactions involving donation of a hydrogen atom, oxygen is reduced to water (H2O) or hydro ...
leads to H2O2 accumulation. Furthermore, in the end process of becoming a toxic
metabolite In biochemistry, a metabolite is an intermediate or end product of metabolism. The term is usually used for small molecules. Metabolites have various functions, including fuel, structure, signaling, stimulatory and inhibitory effects on enzymes, c ...
, it would decompose upon chemical interaction with the reduced sulfur compounds, whose presence is characteristic for the habitat of these bacteria. When grown on the media containing
organic acid An organic acid is an organic compound with acidic properties. The most common organic acids are the carboxylic acids, whose acidity is associated with their carboxyl group –COOH. Sulfonic acids, containing the group –SO2OH, are rel ...
s of the
TCA cycle The citric acid cycle (CAC)—also known as the Krebs cycle or the TCA cycle (tricarboxylic acid cycle)—is a series of chemical reactions to release stored energy through the oxidation of acetyl-CoA derived from carbohydrates, fats, and proteins ...
, the unicellular sulfur bacterium ''M.'' ''bipunctata'' is able to synthesize and store
calcium oxalate Calcium oxalate (in archaic terminology, oxalate of lime) is a calcium salt of oxalic acid with the chemical formula . It forms hydrates , where ''n'' varies from 1 to 3. Anhydrous and all hydrated forms are colorless or white. The monohydrate ...
s inside the cell. This process is possible due to the presence of the high oxaloacetate hydrolase activity in ''M.'' ''bipunctata''. The
oxalate Oxalate (IUPAC: ethanedioate) is an anion with the formula C2O42−. This dianion is colorless. It occurs naturally, including in some foods. It forms a variety of salts, for example sodium oxalate (Na2C2O4), and several esters such as dimethyl o ...
metabolism throughout different cultures was seen through three different enzymes. One of them leads to the formation of
glyoxylate Glyoxylic acid or oxoacetic acid is an organic compound. Together with acetic acid, glycolic acid, and oxalic acid, glyoxylic acid is one of the C2 carboxylic acids. It is a colourless solid that occurs naturally and is useful industrially. Str ...
, which may then enter bio-synthetic reactions. The second way implies oxidation of
oxalate Oxalate (IUPAC: ethanedioate) is an anion with the formula C2O42−. This dianion is colorless. It occurs naturally, including in some foods. It forms a variety of salts, for example sodium oxalate (Na2C2O4), and several esters such as dimethyl o ...
to CO2 via
formate Formate (IUPAC name: methanoate) is the conjugate base of formic acid. Formate is an anion () or its derivatives such as ester of formic acid. The salts and esters are generally colorless.Werner Reutemann and Heinz Kieczka "Formic Acid" in ''Ull ...
, which may be significant in energy metabolism. The third way is oxidation of
oxalate Oxalate (IUPAC: ethanedioate) is an anion with the formula C2O42−. This dianion is colorless. It occurs naturally, including in some foods. It forms a variety of salts, for example sodium oxalate (Na2C2O4), and several esters such as dimethyl o ...
by
oxalate oxidase In enzymology, an oxalate oxidase () is an oxalate degrading enzyme that catalyzes the chemical reaction: :oxalate + O2 + 2 H+ \rightleftharpoons 2 CO2 + H2O2 The 3 substrates of this enzyme are oxalate, O2, and H+, whereas its two products a ...
. Furthermore, ''M.'' ''bipunctata'' was found that reduced sulfur compounds such as H2S were not used by the strains as
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 ...
s, rather, their
oxidation Redox (reduction–oxidation, , ) is a type of chemical reaction in which the oxidation states of substrate change. Oxidation is the loss of electrons or an increase in the oxidation state, while reduction is the gain of electrons or a d ...
was due to interaction with H2O2. This was a main product of O2 reduction in respiration. It is assumed that ''Macromonas bipunctata'', at least in part, is responsible for the metabolism of
organic acid An organic acid is an organic compound with acidic properties. The most common organic acids are the carboxylic acids, whose acidity is associated with their carboxyl group –COOH. Sulfonic acids, containing the group –SO2OH, are rel ...
s and calcium deposition in the form of a calcite crystals. This bacterium recently classified as colorless sulfuric bacterium which has the ability to partially oxidize inorganic sulfur compounds.


Ecology

''M. bipunctata'' lives in several different environments. Other than its communal living in moonmilk formations in certain caves, it was first isolated from a white mat formed in a waste-water. This microorganism is also found as a free-living microbe adapted to high-calcium and high alkaline, freshwater environments.


Biogeochemical significance

''Macromonas bipunctata'' has an indirect connection to the discovery of several antibiotics within the moonmilk formations, but its greatest importance is in its chemical cycling of minerals such as sulfur and calcium in mesophilic environments. This microbe plays a major, holistic role in cycling sulfur through the environment. This bacteria has the ability to precipitate fine crystals of
calcite Calcite is a Carbonate minerals, carbonate mineral and the most stable Polymorphism (materials science), polymorph of calcium carbonate (CaCO3). It is a very common mineral, particularly as a component of limestone. Calcite defines hardness 3 on ...
as a
byproduct A by-product or byproduct is a secondary product derived from a production process, manufacturing process or chemical reaction; it is not the primary product or service being produced. A by-product can be useful and marketable or it can be consid ...
of its activity through calcite inclusions within the cell of the microorganism. It also helps make magnesia crystals and the combination of the two provide the majority of the moonmilk formation that provides a mesophilic environment for several Archaea ad Bacterial phyla that live within the formations.


References


External links


Type strain of ''Macromonas bipunctata'' at Bac''Dive'' - the Bacterial Diversity Metadatabase
{{Taxonbar, from=Q24976895 Comamonadaceae Bacteria described in 1920