Just download the book "Marine Microbiology" from google by DAYA PUBLISHING HOUSE

Hi Brittany, I worked with a E.coli in sediments and used the basic LTB/EC method. However, It was aimed at Total fecal coliforms and E.coli though. Used the same method for water as well since I was comparing water, sediment and shellfish meat bacterial content. You can take a look at it if interested. Sonier et al. 2008. Journal of Applied Ichtyology. 24:678-684.

Best regards,

Remi

I would suggest live/dead staining and cell counts. If you rely on CFU you are limited to only those microbes that are capable of growing on the given medium. In terms of getting cells off of sediment particles, you could try light sonication.

yes I am only interested in enumerating Enterococcus faecalis. And for now have just been spiking in a known concentration of a pure culture to see what I am able to recover.

Hi, if you are only looking at bacteria, you can use differential centrifugation. You first spin down at 2500 rpm to pellet all the larger cells and debris, with the bacteria still in solution. After decanting the solution to a new container, spin down at 10 000 rpm to pellet any bacteria in solution.

Although this is a very basic method, it allows you to study bacteria in any environment, minus some of the background noise. You can also combine diff. centrif. with bead beating - that way you know that most of your extracted DNA will be from bacteria.

I would also suggest using DAPI to stain only viable cells.

Regards,

F