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Deep subsurface mine stalactites trap endemic fissure fluid Archaea, Bacteria, and Nematoda possibly originating from ancient seas.

Borgonie G, Linage-Alvarez B, Ojo A, Shivambu S, Kuloyo O, Cason ED, Maphanga S, Vermeulen JG, Litthauer D, Ralston CD, Onstott TC, Sherwood-Lollar B, Van Heerden E - Front Microbiol (2015)

Bottom Line: Nematoda were clearly identified between these layers confirming that bacteria and nematodes live inside the stalactites and not only in the central straw.Surprisingly, several Bacteria showing highest sequence identities to marine species were identified.Our results indicate stalactites are suitable for biodiversity recovery and act as natural traps for microorganisms in the fissure water long after the water that formed the stalactite stopped flowing.

View Article: PubMed Central - PubMed

Affiliation: Extreme Life Isyensya Gentbrugge, Belgium ; Department of Biotechnology, University of the Free State Bloemfontein, South Africa.

ABSTRACT
Stalactites (CaCO3 and salt) from water seeps are frequently encountered in ceilings of mine tunnels whenever they intersect water-bearing faults or fractures. To determine whether stalactites could be mineralized traps for indigenous fracture water microorganisms, we analyzed stalactites collected from three different mines ranging in depth from 1.3 to 3.1 km. During sampling in Beatrix gold mine (1.4 km beneath the surface), central South Africa, CaCO3 stalactites growing on the mine tunnel ceiling were collected and observed, in two cases, to contain a living obligate brackish water/marine nematode species, Monhystrella parvella. After sterilization of the outer surface, mineral layers were physically removed from the outside to the interior, and DNA extracted. Based upon 16S and 18S rRNA gene sequencing, Archaea, Bacteria, and Eukarya in different combinations were detected for each layer. Using CT scan and electron microscopy the inner structure of CaCO3 and salt stalactites were analyzed. CaCO3 stalactites show a complex pattern of lamellae carrying bacterially precipitated mineral structures. Nematoda were clearly identified between these layers confirming that bacteria and nematodes live inside the stalactites and not only in the central straw. Salt stalactites exhibit a more uniform internal structure. Surprisingly, several Bacteria showing highest sequence identities to marine species were identified. This, together with the observation that the nematode M. parvella recovered from Beatrix gold mine stalactite can only survive in a salty environment makes the origin of the deep subsurface colonization enigmatic. The possibility of a Permian origin of fracture fluids is discussed. Our results indicate stalactites are suitable for biodiversity recovery and act as natural traps for microorganisms in the fissure water long after the water that formed the stalactite stopped flowing.

No MeSH data available.


Related in: MedlinePlus

SEM of Monhystrella parvella inside a CaCO3 stalactite after longitudinal sections with a scalpel. (A) Composite image made of four images taken at different focal points and assembled using Adobe Photoshop. Nematodes (blue) reside in the amorph mass within the confines of the CaCO3 outer stalactite wall (pink). (B) Detail of M. parvella, arrow highlights the fovea. Scale bars: (A,B): 200 μm.
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Figure 5: SEM of Monhystrella parvella inside a CaCO3 stalactite after longitudinal sections with a scalpel. (A) Composite image made of four images taken at different focal points and assembled using Adobe Photoshop. Nematodes (blue) reside in the amorph mass within the confines of the CaCO3 outer stalactite wall (pink). (B) Detail of M. parvella, arrow highlights the fovea. Scale bars: (A,B): 200 μm.

Mentions: Micro CT scanning and SEM of the CaCO3 stalactites that yielded nematodes failed to reveal any apertures in the outermost CaCO3 layer. Micro CT scanning revealed a circular layered internal stalactite structure, laminae, with cavities between the layers themselves populated with several type of different shaped outgrowths mainly, although not always, pointed toward the center (Figures 3A–C, Video 1). All CaCO3 stalactites scanned showed turning laminae more numerous closer to the ceiling than at the bottom. Using CT scan the highest number of laminae counted was 16 layers of partial or complete laminae in the stalactite part near the ceiling, diminishing to a minimum of 8 near the exit of the straw. Four morphologically different outgrowths were identified; the “coralloid,” the “pine,” the “amphora” and the “Chinese fan” (Figures 3D, 4A–F). Nematodes were identified using SEM between these calcified layers (Figures 5A,B) thereby confirming the nematodes also live inside the stalactite and not only in the central tube of the stalactite.


Deep subsurface mine stalactites trap endemic fissure fluid Archaea, Bacteria, and Nematoda possibly originating from ancient seas.

Borgonie G, Linage-Alvarez B, Ojo A, Shivambu S, Kuloyo O, Cason ED, Maphanga S, Vermeulen JG, Litthauer D, Ralston CD, Onstott TC, Sherwood-Lollar B, Van Heerden E - Front Microbiol (2015)

SEM of Monhystrella parvella inside a CaCO3 stalactite after longitudinal sections with a scalpel. (A) Composite image made of four images taken at different focal points and assembled using Adobe Photoshop. Nematodes (blue) reside in the amorph mass within the confines of the CaCO3 outer stalactite wall (pink). (B) Detail of M. parvella, arrow highlights the fovea. Scale bars: (A,B): 200 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4585192&req=5

Figure 5: SEM of Monhystrella parvella inside a CaCO3 stalactite after longitudinal sections with a scalpel. (A) Composite image made of four images taken at different focal points and assembled using Adobe Photoshop. Nematodes (blue) reside in the amorph mass within the confines of the CaCO3 outer stalactite wall (pink). (B) Detail of M. parvella, arrow highlights the fovea. Scale bars: (A,B): 200 μm.
Mentions: Micro CT scanning and SEM of the CaCO3 stalactites that yielded nematodes failed to reveal any apertures in the outermost CaCO3 layer. Micro CT scanning revealed a circular layered internal stalactite structure, laminae, with cavities between the layers themselves populated with several type of different shaped outgrowths mainly, although not always, pointed toward the center (Figures 3A–C, Video 1). All CaCO3 stalactites scanned showed turning laminae more numerous closer to the ceiling than at the bottom. Using CT scan the highest number of laminae counted was 16 layers of partial or complete laminae in the stalactite part near the ceiling, diminishing to a minimum of 8 near the exit of the straw. Four morphologically different outgrowths were identified; the “coralloid,” the “pine,” the “amphora” and the “Chinese fan” (Figures 3D, 4A–F). Nematodes were identified using SEM between these calcified layers (Figures 5A,B) thereby confirming the nematodes also live inside the stalactite and not only in the central tube of the stalactite.

Bottom Line: Nematoda were clearly identified between these layers confirming that bacteria and nematodes live inside the stalactites and not only in the central straw.Surprisingly, several Bacteria showing highest sequence identities to marine species were identified.Our results indicate stalactites are suitable for biodiversity recovery and act as natural traps for microorganisms in the fissure water long after the water that formed the stalactite stopped flowing.

View Article: PubMed Central - PubMed

Affiliation: Extreme Life Isyensya Gentbrugge, Belgium ; Department of Biotechnology, University of the Free State Bloemfontein, South Africa.

ABSTRACT
Stalactites (CaCO3 and salt) from water seeps are frequently encountered in ceilings of mine tunnels whenever they intersect water-bearing faults or fractures. To determine whether stalactites could be mineralized traps for indigenous fracture water microorganisms, we analyzed stalactites collected from three different mines ranging in depth from 1.3 to 3.1 km. During sampling in Beatrix gold mine (1.4 km beneath the surface), central South Africa, CaCO3 stalactites growing on the mine tunnel ceiling were collected and observed, in two cases, to contain a living obligate brackish water/marine nematode species, Monhystrella parvella. After sterilization of the outer surface, mineral layers were physically removed from the outside to the interior, and DNA extracted. Based upon 16S and 18S rRNA gene sequencing, Archaea, Bacteria, and Eukarya in different combinations were detected for each layer. Using CT scan and electron microscopy the inner structure of CaCO3 and salt stalactites were analyzed. CaCO3 stalactites show a complex pattern of lamellae carrying bacterially precipitated mineral structures. Nematoda were clearly identified between these layers confirming that bacteria and nematodes live inside the stalactites and not only in the central straw. Salt stalactites exhibit a more uniform internal structure. Surprisingly, several Bacteria showing highest sequence identities to marine species were identified. This, together with the observation that the nematode M. parvella recovered from Beatrix gold mine stalactite can only survive in a salty environment makes the origin of the deep subsurface colonization enigmatic. The possibility of a Permian origin of fracture fluids is discussed. Our results indicate stalactites are suitable for biodiversity recovery and act as natural traps for microorganisms in the fissure water long after the water that formed the stalactite stopped flowing.

No MeSH data available.


Related in: MedlinePlus