CharonZ - We used agar plates and broth tubes. Growth on the plates was not noticable until yesterday (yay!) by red colonies. Until then we used broth containing 28% NaCl, peptone, casamino acids, FeCl3, Yeast Extract and some other metals and such(I dont have the actual recipe on me, but thats the main components as I recall).
The original cultures were taken from some salt evaporation ponds in Southern California by a CSU Long Beach microbiologist.
We are assuming that the mixed isolates contain the archaean halophiles pretty much becasue of the NaCl concentrations in which they grew. at near saturated levels, I don't know what other organisms could survive. They stained gram negative, and when the plated culture was seen under a dark field microscope, they were spirillium in shape. We ran a gram stain on the new plate growth, but I made the fatal mistake(for the bugs) to put the cells in a drop of regular water, which I am assuming caused lycis, so the gram stain was a bust, and stained gram-postive, but true cells could not be seen, just clumps of matter and such. I am assuming this has something to do with the fact that they don't have PG in their cell walls, but I am not certain.
Half the plates were incubated in 37C dark room, and half the tubes incubated in 37 dark room on a shaker. the others were left in the light at room temp.
The plated growth originated from the 37C dark room.
The new cultures did not contain multiple types of cells, instead it was only the spirillium, and some rod type. The NaCl saturated broth tubes produced the most variety, but the postage stamps were only found in the original culture we made at 28% NaCl. The next saturated cultures only produced long rods, and shorter square rods.
Also, I just streaked some isolation plates from the agar culture that finally grew. The semester is over next week, but I innoculated a large saturated flask in hopes of creating a large culture for further study next semester.
Sorry for the scatter-brained-ness of this post, I just wrote down small things that I'm remembering off the top of my head, and some things come inbetween them.
If you have information, I would love to hear it. I'm presenting the project tomarrow, but I'm still interested in the halophiles. Though the project did not produce any significant results, due to time constraints, I am hoping to further my study of them in the comming months, hopefully with an isolation
