Geochemistry & Ecology of Red Mat Systems (GERMS)

Undergraduate Summer Research Program

 

Week One – Practice

Representative Water Analysis

 

Introduction

Week 1 June 22-24, 2004

 

 

Site Documentation:

 

Luckamute River at Helmick 6/22/04

pH 5.5

temp 19.9 °C

Methods for collection:  we filled two plastic 2 L bottles of moving river water.  

LUCKIAMUTE RIVER COLLECTION SITE

 

 

Field Chemical Test Methods and Results (AccuVac)

 

Methods: For all samples fill a sample cell with 10 ml of sample to be used as a blank for each test.  Fill a beaker with sample, add appropriate reagents, and break off tip of AccuVac bottle in the bottom of the beaker.  Allow to stand for the specified amount of time, and read. 

 

 

 

Results:

mg/L

Total Chlorine  3 min

0.010

Chromium 5 min

0.010

Molybdenum Add 4 drops of CDTA to 40 ml to beaker 5 min

0.200

Nitrate  shake 1 min, 5 min

0.900

Nitrite 15 min

0.003

Sulfate 5 min

0.000

Sulfide (non-Accuvac)  Fill sample cell w/ 25 ml of sample, and another w/ 25 ml of dH2O.  Add 1 ml of Sulfide Reagent 1 to each celll, swirl, add sulfide 5 min

0.000

 

 

 

Filtrate Analysis                                  

Methods:  Set up plastic side-arm flask and ring stand mount, securing it with a clamp.  Wet ring stand mount, and center filter on mount with sterile forceps.  Center 1 L chimney on mount, secure with the blue clamp and fill with 1 L of sample water.  Turn on vacuum, and let run until 1L of water has passed through the filter. 

Removing the filter: After removing chimney, pinch filter with sterile forceps at the apex, and start rolling.

Place roll in sterile 50 mL conical.  Deep freeze conical.

Repeat until all the sample water has been filtered. 

   

FILTERING APPARATUS

                              

 

Representative Pigment and Microscopic Analysis Methods and Results:

 

Methods: In vivo extracts both the pigment and its member bound protein.  Methanol extraction removes only the naked pigment.  The combination of peaks from both absorbency graphs allows us to determine the type of bacteriochlorophyll present and the typical phylogenetic lineage. 

IN VIVO- POD RED LAYER

Results: Peaks

Methanol:

664.0 nm (0.093 ABS)

766.0 nm (0.042 ABS)

 

In vivo:

667.0 nm (0.138 ABS)

733.0 nm (0.105 ABS)

797.0 nm (0.075 ABS)

880.0 nm (0.059 ABS)

 

Bacteriochlorophyll possible:

Bchl a Purple/proteobacteria

Bchl g Gram + Heliobacteria

Bchl d Green Sulfur Bacteria

Bchl c-s Green non-Sulfur Bacteria

METHANOL- POD RED LAYER

 

 

Results:

 

Image – Visible Light

10X

40X

 

 

 

Image – F Fluorescence

10X

40X

 

 

 

Image – R Fluorescence

10X

40X

 

 

 

Discussion:   The microscopy pictures from week 1 were lost and never to be found.