Mycoplasma Alligatoris
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''Mycoplasma alligatoris'' is a species of bacteria in the genus ''
Mycoplasma ''Mycoplasma'' is a genus of bacteria that, like the other members of the class ''Mollicutes'', lack a cell wall, and its peptidoglycan, around their cell membrane. The absence of peptidoglycan makes them naturally resistant to antibiotics ...
''. It is classified in the family Mycoplasmataceae, order Mycoplasma, class Mollicutes, phylum Firmicutes and domain Bacteria. Many organisms of the genus ''Mycoplasma'' are known pathogens in humans and animal species. ''Mycoplasma alligatoris'' is known to elicit a fatal disease with inflammatory characteristics that can cause rapid death of alligators and
caiman A caiman ( (also spelled cayman) from Taíno language, Taíno ''kaiman'') is an alligatorid belonging to the subfamily Caimaninae, one of two primary lineages within the Alligatoridae family (biology), family, the other being alligators. ...
s.


Discovery

''Mycoplasma alligatoris'' was first isolated in a study led by D. R. Brown of six captive alligators showing signs of pneumonia,
polyserositis Familial Mediterranean fever (FMF) is a hereditary inflammatory disorder. FMF is an autoinflammatory disease caused by mutations in the Mediterranean fever (MEFV) gene, which encodes a 781–amino acid protein called pyrin. While all ethnic ...
(inflammation of serous membranes), and multifocal
arthritis Arthritis is a general medical term used to describe a disorder that affects joints. Symptoms generally include joint pain and stiffness. Other symptoms may include redness, warmth, Joint effusion, swelling, and decreased range of motion of ...
. The isolates were obtained from various organs, blood,
synovial fluid Synovial fluid, also called synovia, elp 1/sup> is a viscous, non-Newtonian fluid found in the cavities of synovial joints. With its egg white–like consistency, the principal role of synovial fluid is to reduce friction between the articul ...
, and
cerebrospinal fluid Cerebrospinal fluid (CSF) is a clear, colorless Extracellular fluid#Transcellular fluid, transcellular body fluid found within the meninges, meningeal tissue that surrounds the vertebrate brain and spinal cord, and in the ventricular system, ven ...
and subject to various experimental tests for identification purposes. Primary isolates were cultured in ATCC medium 98 agar containing 105 U penicillin G 1−1, 10 5 U polymyin B 1 −1, 65 mg
cefoperazone Cefoperazone is a third-generation cephalosporin antibiotic, marketed by Pfizer under the name Cefobid. It is one of few cephalosporin antibiotics effective in treating ''Pseudomonas'' bacterial infections which are otherwise resistant to these a ...
1 −1 and 20% (v/v)
fetal bovine serum Fetal bovine serum (FBS) is the most widely used serum-supplement for the ''in vitro'' cell culture of eukaryotic cells. It is commonly utilized in biomedical research, pharmaceutical development, and biomanufacturing due to its ability to suppo ...
in 5% CO2 atmosphere or in ambient air. Isolates were then grown at seven different temperatures in an anaerobic environment, reaching optimum growth at 30 to 34 °C. Cultures were diluted in broth medium then passed through membrane filters of various pore diameters, yielding similar results for all nine isolates. Diluted isolates were then tested for reversion, a method of genetic stabilization unique to some bacteria, in both broth and agar medium. No reversion was observed. Isolates were grown in broth medium with large (20%) and minute (0.2%) amounts of fetal bovine serum as a sterol source. Growth was inhibited in the latter, indicating dependence on an outside sterol source for growth. The
16S rRNA 16S ribosomal RNA (or 16Svedberg, 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 genes coding for it are referred to as ...
gene was sequenced, yielding a unique sequence, suggesting the discovery of a previously unidentified organism. The Brown study in 2001 further investigated the pathogenicity of the newly discovered ''M. alligatoris'' by inoculation of four healthy alligators with the bacteria and one control alligator inoculated with sterile broth. Three of the experimental alligators died within three weeks of inoculation. The surviving alligator tested free of ''M. alligatoris'' after 14 weeks, further supporting the researchers suspicions of the new isolate as the cause of the investigated symptoms.


Preliminary characterization

''Mycoplasma alligatoris'' is a non-motile chemoorganoheterotroph found in American alligators. It has an optimum growth temperature range of 30-34 °C. The DNA sequence for ''M. alligatoris'' type strain A21JP2T was obtained through random
shotgun sequencing In genetics, shotgun sequencing is a method used for sequencing random DNA strands. It is named by analogy with the rapidly expanding, quasi-random shot grouping of a shotgun. The Sanger sequencing#Method, chain-termination method of DNA sequencin ...
, revealing a genome size of 1040–1060 kb, with 89% being coding sequences. The ''M. alligatoris'' genome has a 26% G+C content. DNA analysis identified genes of the
N-acetylneuraminate ''N''-Acetylneuraminic acid (Neu5Ac or NANA) is the predominant sialic acid found in human cells, and many mammalian cells. Other forms, such as N-Glycolylneuraminic acid, may also occur in cells. This residue is negatively charged at physiologi ...
scavenging and catabolism pathway. Biochemical tests including metabolism of glucose,
mannose Mannose is a sugar with the formula , which sometimes is abbreviated Man. It is one of the monomers of the aldohexose series of carbohydrates. It is a C-2 epimer of glucose. Mannose is important in human metabolism, especially in the glycosylatio ...
, lactose, sucrose,
arginine Arginine is the amino acid with the formula (H2N)(HN)CN(H)(CH2)3CH(NH2)CO2H. The molecule features a guanidinium, guanidino group appended to a standard amino acid framework. At physiological pH, the carboxylic acid is deprotonated (−CO2−) a ...
,
aesculin Aesculin, also called æsculin or esculin, is a coumarin glucoside that naturally occurs in the trees horse chestnut (''Aesculus hippocastanum''), California buckeye (''Aesculus californica''), and prickly box (''Bursaria spinosa''). It is al ...
, urea and
phosphatase In biochemistry, a phosphatase is an enzyme that uses water to cleave a phosphoric acid Ester, monoester into a phosphate ion and an Alcohol (chemistry), alcohol. Because a phosphatase enzyme catalysis, catalyzes the hydrolysis of its Substrate ...
activity performed on ''M. alligatoris'' isolates showed that the organism ferments glucose, mannose, lactose, and sucrose and displays phosphatase activity. DNA sequencing analysis revealed genes encoding a complement of
glycosidases In biochemistry, glycoside hydrolases (also called glycosidases or glycosyl hydrolases) are a class of enzymes which catalyze the hydrolysis of glycosidic bonds in complex sugars. They are extremely common enzymes, with roles in nature inclu ...
, which include
hyaluronidase Hyaluronidases are a family of enzymes that catalyse the degradation of hyaluronic acid. Karl Meyer classified these enzymes in 1971, into three distinct groups, a scheme based on the enzyme reaction products. The three main types of hyaluroni ...
s, two
sialidase Exo-α-sialidase (, sialidase, neuraminidase; systematic name acetylneuraminyl hydrolase) is a glycoside hydrolase that cleaves the glycosidic linkages of neuraminic acids: : Hydrolysis of α-(2→3)-, α-(2→6)-, α-(2→8)- glycosidic link ...
s, three β-galactosidases, α-amylase (glycogenase), and two
glycosyltransferase Glycosyltransferases (GTFs, Gtfs) are enzymes ( EC 2.4) that establish natural glycosidic linkages. They catalyze the transfer of saccharide moieties from an activated nucleotide sugar (also known as the "glycosyl donor") to a nucleophilic gl ...
s. ''M. alligatoris'' differs from other species of the Mycoplasma genus by the aforementioned complement of sialidases, which are enzymes of the
hydrolase In biochemistry, hydrolases constitute a class of enzymes that commonly function as biochemical catalysts that use water to break a chemical bond: :\ce \quad \xrightarrowtext\quad \ce This typically results in dividing a larger molecule into s ...
class.


Pathogenicity

Comparisons of the ''M. alligatoris'' genome and that of its nearest known relative, ''Mycoplasma crocodyli'', were analyzed in hopes of understanding the unusually aggressive nature of the ''M.'' ''alligatoris'' species. ''Mycoplasma crocodyli'' is much less virulent than ''M. alligatoris'', lacking the genes for
adhesins Bacterial adhesins are cell-surface components or appendages of bacteria that facilitate adhesion or adherence to other cells or to surfaces, usually in the host they are infecting or living in. Adhesins are a type of virulence factor. Adherence is ...
, variable surface antigens, and
sialidase Exo-α-sialidase (, sialidase, neuraminidase; systematic name acetylneuraminyl hydrolase) is a glycoside hydrolase that cleaves the glycosidic linkages of neuraminic acids: : Hydrolysis of α-(2→3)-, α-(2→6)-, α-(2→8)- glycosidic link ...
s found in ''M. alligatoris.'' Sialidase, in combination with
hyaluronidase Hyaluronidases are a family of enzymes that catalyse the degradation of hyaluronic acid. Karl Meyer classified these enzymes in 1971, into three distinct groups, a scheme based on the enzyme reaction products. The three main types of hyaluroni ...
, is thought to act as a
virulence factor Virulence factors (preferably known as pathogenicity factors or effectors in botany) are cellular structures, molecules and regulatory systems that enable microbial pathogens (bacteria, viruses, fungi, and protozoa) to achieve the following: * c ...
responsible for the pathogenicity of the organism, working together as potential promoters of the expression of
CD95 The Fas receptor, also known as Fas, FasR, apoptosis antigen 1 (APO-1 or APT), cluster of differentiation 95 (CD95) or tumor necrosis factor receptor superfamily member 6 (TNFRSF6), is a protein that in humans is encoded by the ''FAS'' gene. Fas ...
(FasR). CD95 is a protein involved in mediated cell death of eukaryotic cells. In a study done in 2005, researchers used
immunofluorescence Immunofluorescence (IF) is a light microscopy-based technique that allows detection and localization of a wide variety of target biomolecules within a cell or tissue at a quantitative level. The technique utilizes the binding specificity of anti ...
imaging to observe the outcome of ''M. alligatoris'' in vitro on apoptosis by cardiac fibroblasts of alligators. Results of the study showed that M. alligatoris produces an infection that changes the morphology in cardiac fibroblasts, increasing CD95 expression leading to cell death.


Ecological impact

Experimental inoculation of American alligators ('' Alligator mississippienis''), Siamese crocodiles ('' Crocdylus siamensis''), and broad-nosed caimans ('' Caiman latirostris'') with ''M. alligatoris'' yielded growth of the bacteria in all species tested. However, the infected Siamese crocodiles showed no symptoms of disease as seen in alligators and caimans. The commensal relationship observed between ''M. alligatoris'' and ''C. siamensis'' suggests the crocodilian as a potential natural reservoir for ''M. alligatoris.'' The transmission of ''M. alligatoris'' is unknown, but due to its presence in captive alligators, researchers warn against returning captive alligators to their natural habitats, fearing these animals may function as a vector for infecting wild populations. As major predators of their ecosystem, alligators help to control the populations they prey on. The various ecological roles of the alligator also include providing shelter for various animals with their abandoned gator holes. The submerged cavities remain full of water throughout the dry season and provide critical nourishment for an abundance of wildlife such as fish, turtles, birds and insects.


References


External links


Type strain of ''Mycoplasma alligatoris'' at Bac''Dive'' - the Bacterial Diversity Metadatabase
{{Taxonbar, from=Q3868955 Bacteria described in 2001 alligatoris