Long-term oxidant deficiency in Pseudomonas aeruginosa PAO303 results in cells which are non-culturable under aerobic conditions

Binnerup, Svend Jørgen

doi:10.1016/0168-6496(93)90043-7

Cited by 13 publications

(20 citation statements)

References 22 publications

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“…Whereas the low culturability of starved laboratory cultures of Chlorobium BS-1 thus matches the very low cultivation success determined in chemocline samples (Overmann et al, 1992;Manske et al, 2005), nonculturable cells were still capable of maintaining cell integrity over 16 weeks which is significantly longer than for many other bacteria (e.g. Binnerup and Sørensen, 1993;Cho and Kim, 1999).…”

Section: Survival Of Chlorobium Sp Bs-1supporting

confidence: 56%

Large‐scale distribution and activity patterns of an extremely low‐light‐adapted population of green sulfur bacteria in the Black Sea

Marschall

Jogler

Henssge

et al. 2010

Environmental Microbiology

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The Black Sea chemocline represents the largest extant habitat of anoxygenic phototrophic bacteria and harbours a monospecific population of Chlorobium phylotype BS-1. High-sensitivity measurements of underwater irradiance and sulfide revealed that the optical properties of the overlying water column were similar across the Black Sea basin, whereas the vertical profiles of sulfide varied strongly between sampling sites and caused a dome-shaped three-dimensional distribution of the green sulfur bacteria. In the centres of the western and eastern basins the population of BS-1 reached upward to depths of 80 and 95 m, respectively, but were detected only at 145 m depth close to the shelf. Using highly concentrated chemocline samples from the centres of the western and eastern basins, the cells were found to be capable of anoxygenic photosynthesis under in situ light conditions and exhibited a photosynthesis-irradiance curve similar to low-light-adapted laboratory cultures of Chlorobium BS-1. Application of a highly specific RT-qPCR method which targets the internal transcribed spacer (ITS) region of the rrn operon of BS-1 demonstrated that only cells at the central station are physiologically active in contrast to those at the Black Sea periphery. Based on the detection of ITS-DNA sequences in the flocculent surface layer of deep-sea sediments across the Black Sea, the population of BS-1 has occupied the major part of the basin for the last decade. The continued presence of intact but non-growing BS-1 cells at the periphery of the Black Sea indicates that the cells can survive long-distant transport and exhibit unusually low maintenance energy requirements. According to laboratory measurements, Chlorobium BS-1 has a maintenance energy requirement of approximately 1.6-4.9.10(-15) kJ cell(-1) day(-1) which is the lowest value determined for any bacterial culture so far. Chlorobium BS-1 thus is particularly well adapted to survival under the extreme low-light conditions of the Black Sea, and can be used as a laboratory model to elucidate general cellular mechanisms of long-term starvation survival. Because of its adaptation to extreme low-light marine environments, Chlorobium BS-1 also represents a suitable indicator for palaeoceanography studies of deep photic zone anoxia in ancient oceans.

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Section: Survival Of Chlorobium Sp Bs-1supporting

confidence: 56%

Large‐scale distribution and activity patterns of an extremely low‐light‐adapted population of green sulfur bacteria in the Black Sea

Marschall

Jogler

Henssge

et al. 2010

Environmental Microbiology

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“…Also the problems regarding overgrowth can be solved by using dilute media [12]. Finally, the atmosphere of the incubation chambers can be manipulated to provide optimal growth conditions even for anaerobes as demonstrated by Binnerup and Sorensen [13] (Fig. 1).…”

Section: Microcolony Assaysmentioning

confidence: 99%

“…Microcolony counts were obtained on citrate minimal medium (CMM) or CMM supplied with NO; or N,O as electron acceptors, or arginine as a substrate for fermentation. From Binnerup and Sorensen[13].…”

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confidence: 99%

Assessment of metabolic activity of single bacterial cells â new developments in microcolony and dehydrogenase assays

Nybroe¹

1995

FEMS Microbiology Ecology

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Some bacteria lose culturability in natural environments but retain measurable metabolic activity and are thus considered viable. Several techniques have been proposed to determine the activity of nonculturable cells. Due to the considerable physiological heterogeneity of bacterial populations in the environment, it is imperative to apply methods which measure cellular activity at the single cell level. This review focuses on two promising methods: the microcolony assay and the respiration assay based on reduction of 5‐cyano‐2,3‐ditolyl tetrazolium chloride (CTC). In the microcolony assay, viable cells are identified by their ability to perform a limited number of cell divisions and this approach is thus related to conventional culture techniques. Some recent methodological developments of the technique aiming at improving the incubation conditions and the detection of microcolonies are presented. Results obtained by the microcolony technique are used to introduce its advantages and limitations. The CTC‐reduction assay determines a central cellular metabolic activity, but does not measure cell growth. Results of studies using this assay are presented, and it is emphasized that great care should be taken to optimize assay conditions for the studied organisms. Finally, the results obtained by different viability assays are compared. For a specific bacterium, several assays, addressing different aspects of cell metabolism, can provide comparable results suggesting that they provide meaningful viability estimates. On the other hand, the use of viability assays on complex indigenous populations may be ambiguous.

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“…The microcolony-forming units (MCFU) technique allows growth activity in single cells to be followed in mixed populations [24,25] and has recently been combined with FISH in order to quantify the ribosomal RNA content in soil bacteria [16]. In previous studies, we have proposed the MCFU technique to be a useful viability index for AOB in pure cultures [26].…”

Section: Introductionmentioning

confidence: 99%

Quantification of ammonia oxidizing bacteria in soil using microcolony technique combined with fluorescence in situ hybridization (MCFU–FISH)

Hesselsøe¹

2001

FEMS Microbiology Ecology

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A new technique was developed for quantification of viable ammonia oxidizing bacteria (AOB) extracted from soil. Extracted bacteria were grown to microcolony-forming units (MCFU) on membrane filters and selectively stained with 16S rDNA probes using fluorescence in situ hybridization (FISH). The MCFU^FISH technique was used to enumerate a Nitrosomonas europaea population after inoculation to soil and the indigenous AOB populations in native and N-enriched soils. A low extraction efficiency (0.3^3%) of AOB in the native soil indicated predominance of a sessile AOB cell type. In contrast, N enrichments increased extractability of AOB from soil particles. Comparing all soils, a close correlation (R 2 = 0.98) existed between extractable AOB numbers (MCFU^FISH counts) and potential ammonia oxidation activities covering more than three orders of magnitude, which indicated that MCFU^FISH counts reflected the active AOB extracted from the soil. By comparison, the correlation between AOB number estimated by the most-probable number (MPN) method and potential ammonia oxidation was considerably lower (R 2 = 0.56), suggesting that MCFU^FISH counts better reflected the number of active AOB extracted from soil. Hence, the MCFU^FISH technique seems suitable for enumeration of viable and active AOB populations in soils, as long as the extractable population of microcolony-forming AOB is above a current minimum threshold for detection of 10 3^1 0 4 MCFU g 31 dry wt. soil. ß (M. HesselsÖe).Abbreviations : MCFU, microcolony-forming unit; FISH, £uorescence in situ hybridization; AOB, ammonia oxidizing bacteria; MPN, mostprobable number FEMS Microbiology Ecology 38 (2001) 87^95 www.fems-microbiology.org

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Long-term oxidant deficiency in Pseudomonas aeruginosa PAO303 results in cells which are non-culturable under aerobic conditions

Cited by 13 publications

References 22 publications

Large‐scale distribution and activity patterns of an extremely low‐light‐adapted population of green sulfur bacteria in the Black Sea

Large‐scale distribution and activity patterns of an extremely low‐light‐adapted population of green sulfur bacteria in the Black Sea

Assessment of metabolic activity of single bacterial cells â new developments in microcolony and dehydrogenase assays

Quantification of ammonia oxidizing bacteria in soil using microcolony technique combined with fluorescence in situ hybridization (MCFU–FISH)

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Long-term oxidant deficiency in Pseudomonas aeruginosa PAO303 results in cells which are non-culturable under aerobic conditions

Cited by 13 publications

References 22 publications

Large‐scale distribution and activity patterns of an extremely low‐light‐adapted population of green sulfur bacteria in the Black Sea

Large‐scale distribution and activity patterns of an extremely low‐light‐adapted population of green sulfur bacteria in the Black Sea

Assessment of metabolic activity of single bacterial cells â new developments in microcolony and dehydrogenase assays

Quantification of ammonia oxidizing bacteria in soil using microcolony technique combined with fluorescence in situ hybridization (MCFU–FISH)

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Assessment of metabolic activity of single bacterial cells â new developments in microcolony and dehydrogenase assays