It is important to know the contributions of bacteria and fungi to decomposition in connection with both the structure of the food web and the functioning of the ecosystem. However, the extent of the competition between these groups of organisms is largely unknown. The bacterial influence on fungal growth in a soil system was studied by applying three different bacterial inhibitors - bronopol, tylosin and oxytetracycline - in a series of increasing concentrations, and comparing the resulting bacterial and fungal growth rates measured using leucine and acetate-in-ergosterol incorporation, respectively. Direct measurements of growth showed that fungi increased after adding inhibitors; the level of increase in fungal growth corresponded to that of the decrease in bacterial growth, irrespective of the bacterial inhibitor used. Similar antagonistic effects of the bacteria on fungal growth were also found after adding the bacterial inhibitors together with additional substrate (alfalfa or straw plant material). The resulting responses in bacterial and fungal growth indirectly indicated that the negative interaction between fungi and bacteria was mostly attributable to exploitation competition. The results of this study also emphasize the increased sensitivity of using growth-related, instead of biomass-based, measurements when studying bacterial and fungal interactions in soil.
The selective inhibition (SI) technique has been widely used to resolve fungal and bacterial biomass. By studying bacterial growth (leucine/thymidine incorporation) and respiration simultaneously, this study demonstrates that the inhibitors the SI technique is based on do not efficiently or specifically resolve fungal and bacterial contributions to respiration. At concentrations that completely inhibited bacterial growth, the bactericide streptomycin had no influence on the SI technique's respiration measurement, and complete inhibition of bacterial growth using oxytetracycline resulted in marginal respiration reductions. The fungicides captan and benomyl severely inhibited non-target bacterial growth. Cycloheximide did not reduce bacterial growth at moderate concentrations, but the cycloheximide respiration reduction was no higher in a soil with more fungal biomass, casting doubt on its ability to discriminate fungal respiration contribution. Conclusions regarding bacteria and fungi based on the SI technique using these inhibitors are thus compromised. The inhibition of glucose-activated respiration by the bactericide bronopol appeared to correlate with bacterial growth inhibition, however. Bronopol, combined with growth-based techniques, could aid development of a new framework to resolve decomposer ecology in soil.
Pollution-induced community tolerance (PICT) of soil bacteria to the antibiotic tylosin was studied over 95 days. Tylosin was added at increasing concentrations, together with different amounts of alfalfa to study the effects of substrate addition on PICT and bacterial growth in soil. The leucine incorporation technique was used to estimate bacterial growth and as a detection method in the PICT concept. Direct inhibition of the bacterial growth rates, resulting in a dose-response curve, was found above 50 mg of tylosin kg(-1) of soil two days after tylosin addition (IC50 value of 960 mg tylosin kg(-1)). After 10 days of exposure to at least 50 mg of tylosin kg(-1), the PICT was observed and correlated to inhibition of bacterial growth by tylosin. A return of the PICT to control levels was found over time, and after 95 days at 1500 mg of tylosin kg(-1), essentially no PICT was found, as compared to the unpolluted control soil. The return of PICT to pre-exposure levels was not totally reflected in the recovery of bacterial growth. Alfalfa addition did not affect the inhibitory effect of tylosin on bacterial growth rates; neither did it alter the PICT. Since tylosin is relatively rapidly degraded in soil, our results indicate that the PICT will return to prepollution levels when the selective pressure of the toxicant is removed and will thus be a useful technique for monitoring remediation measures.
Pollution-induced community tolerance (PICT) was used to study effects of phenol on soil bacteria. Phenol was added to an agricultural soil in a microcosm experiment. The effects were studied for up to four months. Bacterial growth rates were estimated with the leucine incorporation technique. This technique was also used as detection method for PICT. Changes in community structure were studied using the phospholipid fatty acid (PLFA) pattern. Increased phenol PICT of the bacterial community was found at phenol concentrations above 1 micromol/g wet weight soil. Direct inhibiting effect on bacterial growth rates 1 d after adding phenol was correlated to PICT. Phenol toxicity was reflected by changes in the structure of the bacterial community, although PICT appeared more sensitive than the PLFA method. In soil amended with 1 to 10 micromol phenol/g soil, bacterial growth recovered within one week. In the soil amended with the highest phenol concentration (30 micromol/g soil), bacterial growth rate recovered from total inhibition after 27 d, eventually reaching values six times higher than in the control. However, PICT did not change during the four months the experiment was performed. The specificity of PICT was also studied by examining cotolerance to 2-chlorophenol, 2,4-dichlorophenol, 2,3,6-trichlorophenol, Cu, and Zn. Adding phenol induced cotolerance of the bacterial community to the other phenols, although always at a lover level than to phenol. No cotolerance was found to metals in phenol-polluted soil. We conclude that the PICT concept is a valuable tool in determining phenol toxicity to bacterial communities, especially in situations where bacterial growth has recovered. Cotolerance between different phenols can, however, make interpretations of PICT more complicated.
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