Use of 16S-rRNA to investigate microbial population dynamics during biodegradation of toluene and phenol by a binary culture

Abstract

Interspecies interactions and changes in the rate and extent of biodegradation in mixed culture–mixed substrate studies were investigated. A binary mixed culture of Pseudomonas putida F1 and Burkholderia sp. JS150 degraded toluene, phenol, and their mixture. Both toluene and phenol can serve as sole sources of carbon and energy for both P. putida F1 and strain JS150. To investigate the population dynamics of this system, a fluorescent in‐situ hybridization method was chosen because of its ability to produce quantitative data, its low standard error, and the ease of use of this method. When the binary mixed culture was grown on toluene or phenol alone, significant interactions between the species were observed. These interactions could not be explained by a pure‐and‐simple competition model and were substrate dependent. Strain JS150 growth was slightly inhibited when grown with P. putida F1 on phenol, and P. putida F1 grew more rapidly than expected. Conversely, when the two species were grown together on toluene alone, P. putida F1 was inhibited while strain JS150 was unaffected. During growth of the mixed culture on a combination of toluene and phenol, the interactions were similar to that observed during growth on phenol alone; P. putida F1 growth was enhanced while strain JS150 was unaffected. Because of the observed interspecies interactions, monoculture kinetic parameters were not sufficient to describe the mixed culture kinetics in any experiment. This is one of the first reports of microbial population dynamics in which molecular microbial ecology and mathematical modeling have been combined. The use of the 16S‐rRNA‐based method allowed for observation and understanding of interspecies interactions that were not observable with standard culture‐based methods. These results suggest the need for more investigations that account for both substrate and microbial interactions when predicting the fate of organic pollutants in real systems. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 70: 436–445, 2000.

Characterization of a three bacteria mixed culture in a chemostat: Evaluation and application of a quantitative terminal‐restriction fragment length polymorphism (T‐RFLP) analysis for absolute and species specific cell enumeration

“…The investigation and characterization of mixed cultures is of increasing importance for basic research as well as for ecological, medical and biotechnological applications. These include, for example investigations of interactions of microbial populations (Fredrickson, 1982), differences between specialists and generalists in microbial interactions (Kuenen, 1982), metabolic interactions in mixed culture biofilms (Møller et al, 1998), susceptibility of mixed biofilms to disinfection (Whiteley et al, 2001), quorum sensing in mixed biofilms related to cystic fibrosis (Riedel et al, 2001), fingerprinting of estrogen degrading consortia (Yu et al, 2005), population dynamics in rice field soils (Lueders and Friedrich, 2000), Nisin production by a mixed culture (Shimizu et al, 1999), biodegradation by a binary culture (Rogers et al, 2000), degradation of phenols, benzoates, and phtalates by anaerobic mixed cultures (Levén and Schnürer, 2005). In addition, there are numerous theoretical considerations available on the dynamics of mixed cultures based on mathematical models for up to an infinite number of species available (e.g., Armstrong and McGehee, 1978;Hansen and Hubbell, 1980;Huisman and Weissing, 2001;Lobry and Harmand, 2005;Pilyugin et al, 2004;Smith and Waltman, 1994).…”

“…For understanding underlying mechanisms of community dynamics in ecological, medical as well as in biotechnological applications the quantitative analysis of systems with reduced species number and its specific population dynamics is of great interest. Especially as kinetics derived from pure culture investigations in many cases are not sufficient for mathematical modeling of mixed culture kinetics and dynamics (Rogers et al, 2000).…”

“…Additionally, if dealing with three or more species, selective plating is extremely laborious. For two species mixed culture quantification the use of fluorescent oligonucleotide probes has been reported (Rogers et al, 2000). However, if more than two species in a mixed culture have been investigated this method was only applied for the determination of relative population composition to our knowledge (Chen and Weimer, 2001).…”

Molecular Methods in Biological Systems

“…used to investigate the interactions of a co-culture of Pseudomonas putida F1 and Burkholderia sp. JS150 degrading toluene, phenol, and their mixture (Rogers et al, 2000). Based on their experience with the methods, the authors suggested that the results from this study encourage further investigations wherein substrates and microbial population are monitored simultaneously.…”

Toluene inhibition on an anaerobic reactor sludge in terms of potential activity and composition of acetoclastic methanogens

“…[5][6][7][8][9] Downloaded by [University of Hawaii at Manoa] at 15: 24 27 December 2014 In this study, it is therefore aimed to determine the effect of toluene on an anaerobic sludge taken from a fullscale upflow anaerobic sludge blanket (UASB) reactor in terms of potential activity and composition of acetoclastic methanogens. The SMA tests had been carried out to determine the effect of different toluene concentrations on the potential acetoclastic methanogenic activity of the anaerobic sludge.…”