I think I might be growing pseudomonas? JEN?

[Romans8:28]

As I have mentioned my children and I are experimenting with culturing.
So far most of what we've seen has just been staph. epidermis.
We have been doing some neat experiments on different antibiotics, and antiseptics to see what works to kill bacteria best. I'll post more later, on the final results.
The first test we did were on gram sensitive medium. Phenylethonal agar, and Levine Eosin Methylene Blue agar. (Jen I would have pm this to you but couldn't find you on here sorry! I don't want to bore everyone!) Like I said these grew some staph, (we did the peroxide test) and we did see some yellow cocci but I think that was a contaminate, from checking them etc! The Levine EMB agar only grew one very small round colony on mine and two on my one duaghter. They were round, beta-hemolytic, that's all I could tell.

We also did a broth culture that I let grow a week or so. All of this was in a foam cooler with a heating pad keeping it about 38C. Afterward I tried to culture my broth culture. Both the nutrient agar and the Phenylethonal only grew the staph epidermis., which from what I understand is normal bacteria from our skin! Then today, after 2 days incubation, the Levine EMB agar, which only grows gram neg. bacteria, is growing definately small cultures with a metallic sheen. They are too small yet to determine the hemolysis etc also with the blue in the agar it I can not detect the pyocyanin or blue dye, nor can I detect the grape like or tortia like oder since the mediums kinda stink too!????

We meet with the hospital microbiologist on Tue. so with digital pictures only :) , hope I can ask him also, but..,
This is important because we are pretty sure that I am suffering with a pseudomonas biofilm infection in my kidneys. Isolating this could be very helpful to my doctor, and yes he knows what we're doing, they even gave me the sterile collection cups!
Jen I know you have some experience in this area, I would really like your imput! If you don't want it posted publically please pm me!
(any other doctors of microbiology people out there???)
Thanks in advance,
Shelly

[Katrina]

Repost http://ic-network.com/forum/member.php?u=6178 link to Jen's profile. I will send her a link to this post.

[Romans8:28]

Thanks Katrina:)!

[Sarojini]

Oh, yes, those media stink to high heaven -- generally the grapelike odor from pseudomonas is only detectable when it's actually infecting tissue because of that media stench! :) Sometimes, you can grow special cells and then infect them with pseudomonas and then the stench becomes obvious, since mammalian cell-culture media is "scent-free", if you know what I mean, but this is hard at home, since the cell culture media is really expensive (over $100 per bottle) and cells themselves can cost hundreds if not thousands.

These things can be really tough to culture from yourself at home -- they're tough in the lab too. You may want to let your broth go longer and see. Also, if you have any mechanism of stirring the culture, you may want to try -- some of the bacteria grow better if the culture is aerated a little by mixing or shaking. I'm not sure if pseudomonas is one of them though, since it's been a while since I've done heavy-duty microbiology (I'm more of a mammalian cell biologist now rather than a bacteriologist).

The only other thing I can think of is that if you are experiencing a biofilm problem, the bacteria may be sticking to themselves and you may not be urinating out a lot of them. Certain bacteria can "wall" themselves off, creating nodules or just a bunch of junk -- for example, tuberculosis and certain types of pneumonia can do this in the lung, and it can also happen in the kidney. You may need different media to see them, or it's possible they're not really there.

Other than this; I don't have much to add... like I said, now you're getting into heavy duty micro and it's been a while for me -- still remember basics, but these details are tough. I'm so glad you are exploring this with your kids though -- that is AMAZING and I encourage you to keep them learning with you. It sounds like your home science class is hundreds of times better than what they'd be learning in public school these days (I know I went to public school and I didn't learn half this stuff until college!!) :grouphug:

I may be able to do a little research for you in the next week or so.

[Romans8:28]

Thanks Jen! Since I have another bottle 0f Levine EMB agar yet to put into culture dishes, and about 7 more broth cultures, I may try it again and see. The more I am looking at my microbiology book, and it grows the more it really seems to look like the pseudomonas a. picture and discription! Did you every see pseudomonas a. cutured on this, though I know it may have been a while? If so do you remember what it looked like etc? Right now it is definately metallic colonies all along the culturing loop lines!
The cost of the mammalian cells cultures is out of my reach! Because of the difficulty in actually culturing biofilm infections, I really didn't expect to find anything, especially on cipro!
According to my microbiology book pseudomonas Aeruginosa is an oxidizer and a non-fermenter. It is easy to to aerate the broth culture since it is liquid but can't picture how to aerate or stir the solid agar? :hmm: It may actually have grown because now that I think about it, we areated it without knowing. We tried our hand at gram staining, using a sterile inoculating loop, so had stirred it a few days before actually cuturing. Then stirred it again before using another sterile loop to culture with.
(We actually did pretty well gram staining except when I ordered the gram stain, I was told our microscope was stong enough but not told we needed and oil lense, so it is stained but who can tell the red from the purple:)!)


Thanks for all your input, and for the encouragement! Since we do a lot of unit studies from real books and have been homeschooling many years, we've accumulated a lot of good science, equipment and books. we've had a lot of fun too! The one thing I have loved about homeschooling is the freedom and drive to study all the things I wanted to learn but didn't have time in school! We've had lots of wonderful memories, that right now not only include culturing but also raising an orphaned litter of little baby squirrels. ;)

Thanks again Jen, know we are kinda moving into the deeper side of microbiology, but I am very fasinated with it! (Other then that awful culture media odor :mad: )
Thanks so much for your help :grouphug:

[Katrina]

Shelly, Thank you so much for sharing this...it is so interesting. I find it amazing that you are doing this at home and I thank you for teaching and learning with your kids about this....I truly believe your helping us all with your studies....if for no other way than by spreading around some knowledge and info and interest in science. You are an amazing parent! God Bless!

[Romans8:28]

As always Katrina, thanks for the encouragement, don't feel like an amazing parent, but I appreicaite it! Katrina as I read all the things you suffer with, yet you are always so encouraging and helpful to others here and even with your articles on the remedy find, that is amazing!!
Thanks again for giving me a place to talk out and attempt to figure out some of the mysteries of science. The more we study the more amazed and humbled I am the complexity of creation, and the awesome Creator!!