fileianmac B&B cave Canada initial samples in polybags

Organism: BB1

Gram stain:  +     morphology:  r     width:  1     length:  1.5     microscopic appearance:  chains    

Organism: BB2

Gram stain:  -     morphology:  c     diameter:  1.0     width:  1.0     length:  1.0    

pdylime was made up from peptone dextrose yeast agar with limestone solution added

Organism: ib pdylime yellow

gross culture appearance:  yellow shiny     Gram stain:  -     morphology:  r     width:  0.25     length:  0.5    

Organism: ib pdylime yellow

gross culture appearance:  yellow shiny     Gram stain:  -     morphology:  r     width:  0.25     length:  0.5    

Organism: ib pdylime pink

gross culture appearance:  pink shiny     Gram stain:  -     morphology:  r    

Organism: ib pdylime fringe

gross culture appearance:  pink     Gram stain:  -     morphology:  b     width:  1     length:  2.5    

Organism: ib outerbag 2

gross culture appearance:  no growth    

Organism: ib 4

gross culture appearance:  nocardia     catalase:  k     microscopic appearance:  filaments branching    

Organism: ib wb

gross culture appearance:  spreading     Gram stain:  -     catalase:  k     morphology:  r     width:  0.75     length:  1.5    

Organism: ib pink

gross culture appearance:  pink     catalase:  +    

Organism: ib white

gross culture appearance:  white     Gram stain:  -     catalase:  +     morphology:  r    

Organism: ib 1 edge

gross culture appearance:  cream dull    

Organism: ian swab2

Gram stain:  -     catalase:  -     morphology:  c     diameter:  1.425     width:  1.425     length:  1.425    

Samples were collected on Saturday August 21 2004 from two main sites. Ian McKenzie Group One is in the passage north of the entrance, and is just within sight of dim, filtered daylight. These formations are slightly moist but firm in texture, the walls and surrounding passage are dry . Although it had rained the day before, Formation One was not dripping as before, so no water samples were collected. Formation One measures 23cm x 16cm and is 22cm high. Formation Two has previously been described as 'deflated' in shape, but upon closer inspection may be a small sphere with a second deposition hanging off one side. It is 19cm.x.-l4cm (tapering to 12cm) and 21cm high. Formation Three, the smallest in ~ x 11 cm x 11 cm, and was observed as having a small, hard broccoli-like spray erupting from the bottom entrance side; it is hard to say whether this is the substrate for the 'ball' formation or was formed subsequently. However other broccoli-like formations were observed elsewhere in the cave (see photo).

Organism: group1 outer bag bacteria 1

gross culture appearance:  yellow lobes    

Organism: group1 outer bag bacteria 2

gross culture appearance:  yellow    

1 came from Formation One, at the middle of the side facing towards the centre of the passage (right side as you look towards entrance).

Organism: ib 1 edge

gross culture appearance:  cream dull    

2 came from Formation Two, middle of the side facing away from the entrance (side facing you as you look towards the entrance).

Organism: ib 2 pink

gross culture appearance:  pink     catalase:  +    

Organism: ib 2 white

gross culture appearance:  white     Gram stain:  -     catalase:  +     morphology:  r     width:  1     length:  3    

closest match:

1 Rhodopseudomonas acidophila

75.00% ( 3 / 4 )

Organism: ib 2 yellow

gross culture appearance:  yellow shiny     Gram stain:  -     catalase:  +     morphology:  b     width:  2     length:  3    

Organism: ian-2

Gram stain:  +     morphology:  r     width:  1.0     length:  5.0     microscopic appearance:  chains    

3 came from the bottom of Formation Three.

Organism: ib 3

gross culture appearance:  no growth pdy    

3.1 came from one of the hard broccoli-like nodules that erupt from the bottom of Formation Three towards the entrance side.

Organism: ib 3.1

gross culture appearance:  contaminant    

X came from a ceiling formation that may be a ball formation that is irregular in shape and attached ~ fully to the ceiling, a short distance closer to the entrance.

Organism: ian-x.1

gross culture appearance:  cream shiny     arginine dihydrolase:  -     TSI gas:  -     pH in V-P broth<6:  -     utilization of citrate:  -     tsi:  -     TSI butt:  -     TSI H2S:  -     TSI slant:  -     urease:  -     sellers:  -     nitrate reduced to nitrite:  -     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     glucose oxidase:  -    

closest match:

1 Bacillus insolitus

100.00% ( 4 / 4 )

2 Bacillus badius

100.00% ( 4 / 4 )

3 Vibrio cholerae

75.00% ( 3 / 4 )

Organism: ib X

gross culture appearance:  several bacteria some lobed     

W A came from the wall (right as you face the entrance) closest to Formation Two, where the wall covering is a dry and flaky coating.

Organism: ian-wa

microscopic appearance:  amoeba    

WB came from a shelf below W A sample site, where the wall covering is dry hard nodules.

Organism: ian-wb.1

gross culture appearance:  cream smooth     arginine dihydrolase:  -     pH in V-P broth<6:  -     utilization of citrate:  -     Gram stain:  -     catalase:  -     formation of indole:  -     morphology:  r     urease:  -     voges-proskauer at 25C:  -     sellers:  -     nitrate reduced to nitrite:  -     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     glucose oxidase:  -     width:  1.0     length:  3.0     microscopic appearance:  chains silica chips    

closest match:

1 Enterobacter aerogenes

85.71% ( 6 / 7 )

2 Shigella flexneri

71.43% ( 5 / 7 )

3 Shigella dysenteriae

71.43% ( 5 / 7 )

Organism: ian-wb.2

gross culture appearance:  dull rough     pH in V-P broth<6:  -     utilization of citrate:  -     catalase:  -     oxidase:  +     urease:  -     voges-proskauer at 25C:  -     nitrate reduced to nitrite:  +     microscopic appearance:  difficult    

closest match:

1 Bacillus larvae

80.00% ( 4 / 5 )

2 Moraxella kingii

75.00% ( 3 / 4 )

Organism: ian-wb.3

gross culture appearance:  purple smooth     arginine dihydrolase:  -     TSI gas:  -     pH in V-P broth<6:  -     utilization of citrate:  -     catalase:  +     oxidase:  +     tsi:  -     TSI butt:  -     TSI H2S:  -     TSI slant:  -     urease:  +     voges-proskauer at 25C:  -     sellers:  +     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     glucose oxidase:  -     microscopic appearance:  see bergy    

closest match:

1 Acinetobacter lwoffii

72.73% ( 8 / 11 )

Organism: ian-wb.4

Gram stain:  -     fla:  +     morphology:  r     motility at 25C:  +     width:  0.5     length:  1.5     microscopic appearance:  pointed    

closest match:

1 Acinetobacter lwoffii

72.73% ( 8 / 11 )

2 Rhodocyclus purpureus

71.43% ( 5 / 7 )

Organism: ian-wb.4a

gross culture appearance:  spore     arginine dihydrolase:  -     TSI gas:  -     pH in V-P broth<6:  -     utilization of citrate:  -     Gram stain:  -     catalase:  +     formation of indole:  -     morphology:  r     oxidase:  +     tsi:  -     TSI butt:  -     TSI H2S:  -     TSI slant:  -     urease:  -     voges-proskauer at 25C:  -     sellers:  +     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     glucose oxidase:  -     width:  1.0     length:  3    

closest match:

1 Acinetobacter lwoffii

72.73% ( 8 / 11 )

2 Rhodocyclus purpureus

71.43% ( 5 / 7 )

Organism: ian-wb.4b

gross culture appearance:  large     arginine dihydrolase:  -     TSI gas:  -     pH in V-P broth<6:  -     utilization of citrate:  -     Gram stain:  -     catalase:  +     formation of indole:  -     morphology:  c     oxidase:  +     tsi:  -     TSI butt:  -     TSI H2S:  -     TSI slant:  -     urease:  -     voges-proskauer at 25C:  -     sellers:  +     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     diameter:  3.5     glucose oxidase:  -     width:  3.5     length:  3.5    

closest match:

1 Acinetobacter lwoffii

72.73% ( 8 / 11 )

Organism: ian-wb.5a

gross culture appearance:  spore     arginine dihydrolase:  -     pH in V-P broth<6:  -     utilization of citrate:  -     Gram stain:  +     catalase:  +     formation of indole:  -     morphology:  r     oxidase:  +     urease:  -     voges-proskauer at 25C:  -     sellers:  +     nitrate reduced to nitrite:  -     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     glucose oxidase:  -     width:  1.0     length:  3    

closest match:

1 Acinetobacter lwoffii

75.00% ( 9 / 12 )

2 Shigella sonnei

71.43% ( 5 / 7 )

3 Rhodocyclus purpureus

71.43% ( 5 / 7 )

Organism: ian-wb.5b

gross culture appearance:  large     arginine dihydrolase:  -     pH in V-P broth<6:  -     utilization of citrate:  -     Gram stain:  +     catalase:  +     formation of indole:  -     morphology:  c     oxidase:  +     urease:  -     voges-proskauer at 25C:  -     sellers:  +     nitrate reduced to nitrite:  -     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     diameter:  3.5     glucose oxidase:  -     width:  3.5     length:  3.5    

closest match:

1 Acinetobacter lwoffii

75.00% ( 9 / 12 )

Organism: ian-wb.6

gross culture appearance:  mixed cream dull     arginine dihydrolase:  -     TSI gas:  -     pH in V-P broth<6:  -     utilization of citrate:  -     Gram stain:  +     catalase:  k     formation of indole:  -     morphology:  r     oxidase:  +     tsi:  -     TSI butt:  -     TSI H2S:  -     TSI slant:  -     urease:  -     voges-proskauer at 25C:  -     sellers:  -     nitrate reduced to nitrite:  -     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     glucose oxidase:  -     width:  2.0     length:  3.5    

closest match:

1 Bacillus badius

85.71% ( 6 / 7 )

2 Bacillus Sphaericus

71.43% ( 5 / 7 )

3 Bacillus Schlegelii

71.43% ( 5 / 7 )

Organism: ib wb 1

gross culture appearance:  spreading     Gram stain:  -     catalase:  k     morphology:  r     width:  0.75     length:  1.5    

Group Two is in a passage south of the entrance and beyond any daylight penetration; the passage here is generally wet and in fact the floor had a small amount of running water, although none of the formations have ever been observed dripping. There is extensive moist moonmilk in this area. Formations in Group Two tend to be quite soft and moist in texture. Group Two formations seem to be more obviously coated with a second (subsequent to sphere) moonmilk deposit. Formation Four is 7cm x 6cm and 7cm high. Formation Five is 9cm x 9cm and 13cm high including a moonmilk 'beard'. Formation Six is irregular in shape and attached to the ceiling with a maximum dimension (length) of 19cm

Organism: group2 outer bag fungus

gross culture appearance:  fungus spores black    

Organism: group2 outer bag bacteria 1

gross culture appearance:  yellow    

Organism: group2 outer bag bacteria 2

gross culture appearance:  yellow lobes    

4 came from Formation Four, at the middle of the side.

Organism: ib 4 mixed organisms

gross culture appearance:  fungi and bacteria    

Organism: ib 4

gross culture appearance:  nocardia     catalase:  k     microscopic appearance:  filaments branching    

5 came from Formation Five, from the bottom.

Organism: ib 5

gross culture appearance:  contaminant    

6 came from Formation Six, similar in shape to formation X in Group One.

Organism: ib 6

gross culture appearance:  no growth pdy    

Organism: ian-6a

Gram stain:  -     morphology:  r     width:  1.0     length:  5.0    

Organism: ian-6b

Gram stain:  -     morphology:  c    

Organism: ian-6c

width:  3.0     length:  10.0     microscopic appearance:  arthrospores long chains rectangular     

W2 came from the right hand wall in the photo, where there is extensive moonmilk.

Organism: ib w2

gross culture appearance:  Geotrichum     catalase:  +     microscopic appearance:  filaments    

Organism: ian-w2.1

gross culture appearance:  tiny cream     arginine dihydrolase:  -     TSI gas:  -     pH in V-P broth<6:  -     utilization of citrate:  -     tsi:  -     TSI butt:  -     TSI H2S:  -     TSI slant:  -     urease:  -     sellers:  -     nitrate reduced to nitrite:  -     sellers slant:  -     sellers butt:  -     sellers band:  -     pyorubrin:  -     sellers nitrogen gas:  -     glucose oxidase:  -    

closest match:

1 Bacillus insolitus

100.00% ( 4 / 4 )

2 Bacillus badius

100.00% ( 4 / 4 )

3 Vibrio cholerae

75.00% ( 3 / 4 )

end