Lysis of Vibrio costicolus by Osmotic Shock

Christian, J. H. B.; Ingram, M.

doi:10.1099/00221287-20-1-32

Cited by 22 publications

(12 citation statements)

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“…V. costicola can use the energy of respiration for proton efflux at alkaline pHo. When the bacteria were incubated anaerobically in a solution containing 1.0 M KCI to prevent lysis (9) and to maintain equal osmolarity to the growth medium, an oxygen pulse was followed by a rapid acidification of the medium (Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

“…A role for antiport activity in pH regulation of the cytosol can also explain the broad pH range for optimal growth, extending to the alkaline extreme of pH 9.0. Vibrio costicola is a moderately halophilic bacterium which requires 1 M NaCl for optimal growth (13) and lyses in media of low osmotic strength (9). Lysis can be prevented by salts other than NaCl, but there is a specific Na+ requirement for carrier-mediated transport (24).…”

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Proton motive force and Na+/H+ antiport in a moderate halophile

Hamaide¹,

Kushner²,

Sprott³

1983

J Bacteriol

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The influence of pH on the proton motive force of Vibrio costicola was determined by measuring the distributions of triphenylmethylphosphonium cation (membrane potential, A&) and either dimethyloxazolidinedione or methylamine (osmotic component, ApH). As the pH of the medium was adjusted from 5.7 to 9.0, the proton motive force steadily decreased from about 170 to 100 mV. This decline occurred, despite a large increase in the membrane potential to its maximum value at pH 9.0, because of the loss of the pH gradient (inside alkaline). The cytoplasm and medium were of equal pH at 7.5; membrane permeability properties were lost at the pH extremes of 5.0 and 9.5. Protonophores and monensin prevented the net efflux of protons normally found when an oxygen pulse was given to an anaerobic cell suspension. A Na+/H+ antiport activity was measured for both Na+ influx and efflux and was shown to be dissipated by protonophores and monensin. These results strongly favor the concept that respiratory energy is used for proton efflux and that the resulting proton motive force may be converted to a sodium motive force through Na+/H+ antiport (driven by A,). A role for antiport activity in pH regulation of the cytosol can also explain the broad pH range for optimal growth, extending to the alkaline extreme of pH 9.0. Vibrio costicola is a moderately halophilic bacterium which requires 1 M NaCl for optimal growth (13) and lyses in media of low osmotic strength (9). Lysis can be prevented by salts other than NaCl, but there is a specific Na+ requirement for carrier-mediated transport (24).Understanding the specific requirement for Na+ in transport requires knowledge of the electrochemical ion gradients in V. costicola.A transmembrane proton motive force (APIH+) may be established in various microorganisms by proton efflux by using respiratory energy, light energy in photosynthetic organisms or extreme halophiles, or the energy from ATP hydrolysis (15,36), resulting in a membrane potential (A+i, interior negative) and an osmotic component (ApH, interior alkaline) such that in millivolts, AP.H+ = A+ -60 ApH, at 30°C (30).Energy may be released from the osmotic component by proton-symport mechanisms in medium more acidic than the internal pH (pHi); at a higher pH of the medium (pHo), the cytosol is more acidic than the medium, so A4 must compensate for the inverse pH gradient.In certain bacteria, a sodium motive force may be formed by several mechanisms, resulting from the outward-directed movement of Na+. t National Research Council of Canada paper no. 22690.First, the energy of the proton motive force may be converted to a sodium motive force through Na+/H+ antiport activity as found in Alteromonas haloplanktis (33), Halobacterium halobium (26), alkalophilic bacilli (29), Mycoplasma mycoides (6), and others, including Escherichia coli (4, 44) (for a review, see reference 25). In addition to a role in energy coupling, antiport activity may be involved in regulation of cytosolic pH (34). Second, in Klebsiella aerogenes, efflux of Na+ th...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Proton motive force and Na+/H+ antiport in a moderate halophile

Hamaide¹,

Kushner²,

Sprott³

1983

J Bacteriol

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“…It has also been shown (Christian & Ingram, 1959) that the osmotic sensitivity of both halophilic and non-halophilic…”

Section: J H B Christian and M Ingrammentioning

confidence: 99%

The Freezing Points of Bacterial Cells in Relation to Halophilism

Christian

Ingram

1959

Journal of General Microbiology

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SUMMARY:The hypothesis that halophilic bacteria achieve a high degree of salt tolerance by exclusion of much of the external solutes of the growth medium has been tested. Organisms were grown in media containing from 1 to 4~ salts. For both halophilic and non-halophilic bacteria the freezing points of the cells were close to those of the media in which they were grown. It is concluded that halophilic bacteria do not maintain the intracellular aqueous phase at a water activity greater than in the surrounding growth medium.One possible explanation for the phenomenon of bacterial halophilism is that the solutes in the aqueous phase within halophilic cells are much less concentrated than in the surrounding medium (Robinson, Gibbons & Thatcher, 1952). Such a system might be maintained, apart from mechanisms of accumulation and excretion, by a membrane impermeable to external solutes and a rigid cell boundary which resists plasmolysis. To examine this hypothesis, the freezing-point depressions of bacterial suspensions and of their growth media have been determined and compared. The bacteria included both halophilic and non-halophilic strains grown in media containing various concentrations of salts. No appreciable differences in the osmotic status of these two types of bacteria were observed. lMETHODSThe four halophilic strains tested were Micrococcus halodenitri$cans, Vibrio costicolus, Halobacterium halobium and Sarcina littoralis from the collection of Dr N. E. Gibbons, National Research Council, Ottowa. Other cultures used were Micrococcus lysodeikticus (NCTC 2665) each of proteose peptone (Difco) and tryptone (Difco) with additions of NaCI, or of NaCl and KC1 as required. The initial pH value was 7.0 in all media. These media were solidified by 2 % (w/v) Bacto agar.To decrease the amount of contaminating medium in suspensions of organisms, the bacteria were grown on solid media in Petri dishes. An inoculum

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“…One of the characteristics of this organism is its osmotic fragility, which causes the cells to lyse and become rapidly inactivated in distilled water (Lee, 1972;Yanagizawa, 1964) . Several investigations have been reported with regard to the lytic phenomenon of halophilic bacteria (Buckmire and MacLeod, 1965;Christian and Ingram, 1959;MacLeod and Matula, 1962;Unemoto, 1979) . Lysed cells generally leak intracellular materials, while maintaining its morphology.…”

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Bleb Formation on the Cell Envelope of Osmotic Shocked Cells of Vibrio parahaemolyticus.

et al. 1999

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Vibrio parahaemolyticuscells are known to lyse in distilled water on account of their osmotic fragility. When V. parahaemolyticus cells were subjected to osmotic shock, blebs were formed on cell envelopes, mainly at the septa of dividing cells and polar regions. Some of the blebs appeared to be detached from the cell surface to form vesicles. Our results indicate that surface blebs and vesicles formed by osmotic shock mainly involve the outer membrane . Although the cytoplasmic membrane itself is not released, its permeability barrier is disrupted.

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Lysis of Vibrio costicolus by Osmotic Shock

Cited by 22 publications

References 10 publications

Proton motive force and Na+/H+ antiport in a moderate halophile

Proton motive force and Na+/H+ antiport in a moderate halophile

The Freezing Points of Bacterial Cells in Relation to Halophilism

Bleb Formation on the Cell Envelope of Osmotic Shocked Cells of Vibrio parahaemolyticus.

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