Different methods for extracting bacteria from freshwater sediment and a simple method to measure bacterial production in sediment samples

被引:49
作者
Furtado, ALD
Casper, P
机构
[1] Inst Freshwater Ecol & Inland Fisheries, Dept Limnol Stratifeld Lakes, D-16775 Neuglobsow, Germany
[2] Univ Fed Rio de Janeiro, Lab Limnol, Inst Biol, Dept Ecol, BR-21941590 Rio De Janeiro, Brazil
关键词
bacteria extraction; centrifugation; freshwater sediments; leucine uptake; sonication;
D O I
10.1016/S0167-7012(00)00163-9
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The efficiency of different treatments was tested to extract bacterial cells from freshwater sediment samples. The influence of sonication, density gradient centrifugation, fixation by formalin and centrifugation speed on bacterial recovery was investigated. The method developed by Smith and Azam [Mar. Microb. Food Webs 6 (1992) 107] to measure microbial activity on bacterioplankton (H-3-leucine incorporation), was also evaluated in sediment samples. After 1 min of sonication bacterial abundance was reduced by about 47% in diluted sediments with tetrasodium pyrophosphate. With the addition of Percoll after sonication, bacterial counts were not significantly different (P < 0.05). Fixation by formalin increased bacterial counts using sonication. However, higher bacterial abundance was estimated in non-sonicated samples. Bacterial abundance in samples centrifuged at 7000 X g with and without Percoll was not significantly different (P < 0.05). Highest bacterial abundance was obtained after centrifugation at low speed (750 X g). Bacterial abundance decreased with higher centrifugation speed (750, 1500 and 3000 X g), the difference, however, was not significant. Bacterial production ranged from 0.10 mu g C cm(-3) d(-1) in autoclaved sediment to 0.27 mu g C cm(-3) d(-1) in untreated sediment. The radioactivity measured in controls of both untreated and autoclaved sediment was high (70 and 91%, respectively), indicating a high level of leucine adsorption in sediment particles. In contrast, radioactivity in control samples previously centrifuged was markedly lower (6%). Despite the high values of radioactivity in the controls, bacterial production in untreated sediment was significantly higher than in centrifuged sediment (P < 0.05). (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:249 / 257
页数:9
相关论文
共 29 条
[11]   ETHANOL-EXTRACTABLE SUBSTRATE POOLS AND THE INCORPORATION OF THYMIDINE, L-LEUCINE, AND OTHER SUBSTRATES BY BACTERIOPLANKTON [J].
HOLLIBAUGH, JT ;
WONG, PS .
CANADIAN JOURNAL OF MICROBIOLOGY, 1992, 38 (07) :605-613
[12]   ASSESSMENT OF [H-3] THYMIDINE INCORPORATION INTO DNA AS A METHOD TO DETERMINE BACTERIAL PRODUCTIVITY IN STREAM BED SEDIMENTS [J].
KAPLAN, LA ;
BOTT, TL ;
BIELICKI, JK .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1992, 58 (11) :3614-3621
[13]   USE OF FLUOROCHROMES FOR DIRECT ENUMERATION OF TOTAL BACTERIA IN ENVIRONMENTAL-SAMPLES - PAST AND PRESENT [J].
KEPNER, RL ;
PRATT, JR .
MICROBIOLOGICAL REVIEWS, 1994, 58 (04) :603-615
[14]   EVALUATION OF METHODS FOR EXTRACTION OF BACTERIA FROM SOIL [J].
LINDAHL, V ;
BAKKEN, LR .
FEMS MICROBIOLOGY ECOLOGY, 1995, 16 (02) :135-142
[15]   Benthic mineralization rates at two locations in the southern North Sea [J].
Osinga, R ;
Kop, AJ ;
Duineveld, GCA ;
Prins, RA ;
VanDuyl, FC .
JOURNAL OF SEA RESEARCH, 1996, 36 (3-4) :181-191
[16]  
Prieme A, 1996, FEMS MICROBIOL ECOL, V21, P59
[18]   USE OF A FLUORESCENT REDOX PROBE FOR DIRECT VISUALIZATION OF ACTIVELY RESPIRING BACTERIA [J].
RODRIGUEZ, GG ;
PHIPPS, D ;
ISHIGURO, K ;
RIDGWAY, HF .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1992, 58 (06) :1801-1808
[19]   THE ECOLOGY OF SEDIMENT BACTERIA IN LAKES AND COMPARISONS WITH OTHER AQUATIC ECOSYSTEMS [J].
SCHALLENBERG, M ;
KALFF, J .
ECOLOGY, 1993, 74 (03) :919-934
[20]   SOLUTIONS TO PROBLEMS IN ENUMERATING SEDIMENT BACTERIA BY DIRECT COUNTS [J].
SCHALLENBERG, M ;
KALFF, J ;
RASMUSSEN, JB .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1989, 55 (05) :1214-1219