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standardization of environmental data and information - International ...

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Area Parameters Instruments Levels <strong>of</strong> observation<br />

Temperature, salinity<br />

<strong>and</strong> light transmission<br />

pr<strong>of</strong>iles <strong>and</strong> water<br />

sampling<br />

CTD meter fitted with<br />

light<br />

transmissometer<br />

<strong>and</strong> altimeter<br />

Entire water column close to<br />

bottom; water sampling at above<br />

depths<br />

1 Sample<br />

collection<br />

2.2. Microbiological <strong>and</strong> biochemical analyses<br />

By P.A. Loka Bharathi, Shanta Nair, Ch<strong>and</strong>ralata Raghukumar, C. Moh<strong>and</strong>ass, NIO<br />

Parameter Variables Methodology for analyses<br />

Sampler<br />

Core length<br />

Sections<br />

2a Microbiology Direct bacterial<br />

counts<br />

2b<br />

Plate counts <strong>of</strong><br />

CFUs<br />

Multiple corer, box corer<br />

30 centimetres<br />

0-2, 2-4, 4-6, 6-8, 8-10, 10-15, 15-20, 20-25, 25-30 cm<br />

Samples directly extruded into sterile plastic bags for<br />

microbiological <strong>and</strong> biochemical analyses<br />

Bacteria enumerated following Hobbie et al. 3 : aliquot <strong>of</strong><br />

sediment sample diluted <strong>and</strong> fixed with filter-sterilized<br />

buffered formalin to give final concentration <strong>of</strong> 2 percent;<br />

fixed sample ultrasonicated at 20 hertz for 30 seconds,<br />

filtered through 0.22-? black nuclepore filter <strong>and</strong> stained<br />

with 0.01% acridine orange for 3 minutes. Bacterial cells<br />

counted using epifluorescence microscopy; average <strong>of</strong> 200<br />

cells/sample counted, with counts expressed as<br />

numbers/gram dry weight. Sediment dried at 60? C for 24<br />

hours.<br />

Bacteria: ¼-strength Zobell Marine Agar<br />

Fungi: 1/5-strength Malt Extract Agar fortified with<br />

streptomycin <strong>and</strong> penicillin to prevent bacterial growth<br />

Incubating temperature: 5-10? C for 10-12 days<br />

Serially diluted sample used as inoculum; numbers expressed<br />

as CFU/g dry wt -1<br />

536<br />

INTERNATIONAL SEABED AUTHORITY

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