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Journal of Bacteriology  1963;85(4):918-926.
Bateman, J. B. (U.S. Army Biological Laboratories, Fort Detrick, Frederick, Md.) and F. Elizabeth White. Effect of relative humidity on the survival of Serratia marcescens in concentrated glycerol and sucrose solutions. J. Bacteriol. 85:918–926. 1963.—The effects of sucrose and glycerol upon the ability of Serratia marcescens to grow when restored to a normal medium after exposure to solutions of these substances were examined, with special attention to the prevailing thermodynamic activity of water in these solutions as a factor of supposed primary importance in influencing survival or death of cells. The data were notable for the absence of any zones of instability such as those found when the water activity is changed by exposure of washed cells to water vapor at controlled relative humidities (RH). The cells survived indefinitely at room temperature in concentrated sucrose solutions; in glycerol solutions of equilibrium RH values from 20 to 90, the first-order decay constants were about 0.03 to 0.1 hr−1. These results, considered together with the contrasting phenomenon of narrow lethal humidity zones found in vapor-phrase equilibration experiments, were explained generally in terms of competitive interactions involving concentrated intrinsic and adventitious solutes, the cell water, and the organized structures of the cell, whose integrity was considered to depend ultimately upon the net effect of these various interactions.
PMCID: PMC278245  PMID: 14044963
2.  Survival of Microorganisms in Laundered Polyester-Cotton Sheeting1 
Applied Microbiology  1973;25(3):431-435.
The effects of wash-water temperature, cold-water or regular detergent, wash-cycle design, drying, and drying temperature on survival of four microorganisms on polyester-cotton sheeting were examined. Escherichia coli T3 bacteriophage survived washing at 24, 35, 46, and 57 C, but not at 68 C. Serratia marcescens survived only the lowest three wash temperatures. Levels of residual Staphylococcus aureus were diminished at the highest two wash temperatures, but survival was substantial even at 68 C. Counts of Bacillus stearothermophilus spores were not altered appreciably by wash temperature. Type of detergent had no practical effect on observed counts. The regular wash cycle was significantly more efficient in removal of microorganisms than the permanent-press cycle. Counts, especially of the bacteriophage and the gramnegative bacterium, were decreased by drying; after drying, the effects of wash-water temperature on S. aureus and B. stearothermophilus were not significantly different. Microorganisms were transferred from inoculated to sterilized sheeting during laundering. The public health significance of these observations is discussed.
PMCID: PMC380823  PMID: 4572894
3.  Relative Humidity and the Killing of Bacteria 
Applied Microbiology  1961;9(6):567-571.
The viability of washed moist cells of Serratia marcescens after storage has been measured in relation to variations in the prior treatment of the cells and in conditions of storage. The factors considered were: (i) water content during storage; (ii) method of arriving at water content (partial drying in vacuum or freeze-drying and addition of water); (iii) presence or absence of air during storage.
Increasingly rapid decay occurs as the water content at which the cells are stored is diminished from above 90% to 20 or 30% (“critical” water content). It occurs in presence or absence of air and it occurs whether the final water content is approached by removal of water from wet cells or by addition of water to freeze-dried cells.
The rate of decay during storage at 20 to 30% water is somewhat diminished by the presence of air (“protective” effect of air).
As the water content is further reduced to less than 10%, the stability of cells stored in a vacuum approaches that of wet cells. In presence of air the reverse is true: the stability decreases until at less than 1% water, the decay rate is about as great as at the “critical” water content (“toxic” effect of air).
Particularly rapid decay of S. marcescens at the “critical” water content has escaped attention in aerosol studies because accurate control of relative humidity (RH) in this region, RH 94 to 99%, is virtually impossible in such studies. On the other hand, values of decay rates referred to measured water contents are quite unreliable in the 20 to 80% RH zone because the corresponding variation of water content is too small to measure reliably. Thus data of the kind reported in this paper cannot be directly compared to the published results of studies of air-borne bacteria, although they are relevant to the practical question of air-borne infection in humid atmospheres.
PMCID: PMC1057789  PMID: 13865722
4.  Effects of Oxygen on Aerosolized Serratia marcescens 
Applied Microbiology  1965;13(5):781-787.
Suspensions of Serratia marcescens (ATCC strain 14041) in water were aerosolized in a rotating drum in the presence of various concentrations of oxygen. The colony-forming ability of aerosolized organisms was rapidly destroyed by contact with 0.25% or more oxygen at 40% relative humidity (RH) and 25 C, but was almost unimpaired for at least 5 hr in nitrogen containing not more than 10 ppm of oxygen. Completely hydrated organisms were insensitive to oxygen at pressures up to 100 psi for 4 hr. No loss in viability occurred in aerosols of washed cells in air at 97% RH. It is proposed that dehydration of the aerosolized cell results in sensitization to lethal effects of oxygen, but is not the primary cause of death. Mn++, Co++, glycerol, and thiourea enhanced the biological stability of aerosols in air. Numerous similarities between the effects of oxygen in this system and in systems using freeze-dried or irradiated organisms or cell-free enzymes support the hypothesis that closely related mechanisms are involved.
PMCID: PMC1058343  PMID: 5325941
5.  Growth and survival of Serratia marcescens under aerobic and anaerobic conditions in the presence of materials from blood bags. 
Journal of Clinical Microbiology  1993;31(7):1826-1830.
Several patients receiving blood transfusions during the summer of 1991 developed bacteremia after the transfusion. In all cases, the infection was caused by Serratia marcescens. The same strain of Serratia marcescens was isolated from the patients and from the outer surface of unfilled blood bags. The transport containers for the blood bags were made anoxic by using a catalyst in order to prevent microbial growth. The survival and growth of S. marcescens K202, which was isolated from the blood bags, was studied at different oxygen concentrations in deionized water containing materials derived from the blood bags. The rate of survival and growth of S. marcescens was highest under anaerobic conditions, in which growth occurred with all materials and even in deionized water alone. In contrast, S. marcescens did not survive in control cultures under semi-anaerobic and aerobic conditions. Growth was observed, however, under both aerobic and semi-anaerobic conditions in the presence of each of the tested blood bag materials. These findings indicate that the conditions in the transport containers for the blood bags were favorable for the survival and growth of S. marcescens.
PMCID: PMC265640  PMID: 8349760
6.  Effects of sorption on biological degradation rates of (2,4-dichlorophenoxy) acetic acid in soils. 
Three mathematical models were proposed to describe the effects of sorption of both bacteria and the herbicide (2,4-dichlorophenoxy)acetic acid (2,4-D) on the biological degradation rates of 2,4-D in soils. Model 1 assumed that sorbed 2,4-D is not degraded, that only bacteria in solution are capable of degrading 2,4-D in solution, and that sorbed bacteria are not capable of degrading either sorbed or solution 2,4-D. Model 2 stated that only bacteria in the solution phase degrade 2,4-D in solution and that only sorbed bacteria degrade sorbed 2,4-D. Model 3 proposed that sorbed 2,4-D is completely protected from degradation and that both sorbed and solution bacteria are capable of degrading 2,4-D in solution. These models were tested by a series of controlled laboratory experiments. Models 1 and 2 did not describe the data satisfactorily and were rejected. Model 3 described the experimental results quite well, indicating that sorbed 2,4-D was completely protected from biological degradation and that sorbed- and solution-phase bacteria degraded solution-phase 2,4-D with almost equal efficiencies.
PMCID: PMC373553  PMID: 3994366
7.  Preservation of Serratia marcescens by High-Vacuum Lyophilization 
Applied Microbiology  1966;14(4):561-567.
Water-washed Serratia marcescens (ATCC strain 14041) cells were lyophilized in an all-glass system capable of evacuation to pressures of less than 5 × 10-6 torr. Lyophilization at the lowest pressures resulted in 50 to 65% survival for unstabilized washed organisms compared with 10 to 20% when the cells were lyophilized at pressures of about 2.5 × 10-2 torr. At the latter pressures, 45 to 65% survival was obtained when NaCl or Naylor-Smith stabilizer was added to the cell suspensions before lyophilization. However, the stabilizers failed to increase significantly the levels of survival compared with water suspension when cells were lyophilized at pressures less than 10-5 torr. The high survival rates obtained by the high-vacuum technique may be attributed to the reduction of traces of molecular oxygen which has been reported to be destructive of the dried bacteria. Survival of unstabilized dried S. marcescens after 1-day storage increased markedly with decreasing sealing pressure. Under the highest vacuum attained, survival of the dried bacteria was not impaired by storage for up to 1 month at Dry Ice temperatures; at higher temperatures, viability losses occurred. Exposure of the dried unstabilized bacteria to dry air resulted in rapid viability loss. The inactivation could be stopped almost immediately by evacuation to pressures of less than 10-5 torr, but the evacuation failed to reverse the viability losses that occurred during exposure.
PMCID: PMC546782  PMID: 5332950
8.  Sorption of Cadmium by Microorganisms in Competition with Other Soil Constituents † 
The fate of cadmium in soil is influenced to a great extent by microbial activity. Microorganisms were compared with abiotic soil components for their ability to sorb Cd from a liquid medium. When the same amount (on a dry weight basis) of bacterial cells (Serratia marcescens and Paracoccus sp.), clay (montmorillonite), or sand was separately incubated in 0.05 M phosphate buffer, pH 7.2, containing 10 ppm of Cd (10 μg/ml), bacterial cells removed the largest quantity of Cd. Dead cells sorbed much more Cd from the medium than live cells. A comparative study of Cd removal from the medium by seven soil bacteria and four fungi did not indicate appreciable differences. With increasing microbial biomass, the relative efficiency of 0.1 M NaOH as an extractant of sorbed Cd increased, whereas the extraction efficiency of 0.005 M DTPA (diethylenetriaminepentaacetic acid) decreased. It appeared that NaOH and DTPA extracted different chemical forms of Cd. This assumption was supported by vastly different correlation coefficients in the relative amount of Cd extracted by the two solvents.
PMCID: PMC244178  PMID: 16346002
9.  Relationship Between Atmospheric Temperature and Survival of Airborne Bacteria 
Applied Microbiology  1970;19(2):245-249.
Effects of temperatures ranging from −40 to 49 C on the behavior of airborne Serratia marcescens, Escherichia coli, and Bacillus subtilis var. niger were investigated. Aerosol decay rates of B. subtilis spores were not significantly affected by the temperature and remained approximately constant within the temperature range studied. The survival of airborne S. marcescens and E. coli was closely related to the temperature. An increase in temperature from −18 to 49 C resulted in a progressive increase of the biological death rate, and the relationship between the biological death rate and the temperature appeared to be linear. An increase in temperature from 24 to 49 C resulted in significantly reduced aerosol recoveries of the two vegetative organisms. At −40 C, the aerosol recovery of all three agents was consistently lower than at −18 to 24 C.
PMCID: PMC376659  PMID: 4985428
10.  Size and UV Germicidal Irradiation Susceptibility of Serratia marcescens when Aerosolized from Different Suspending Media 
Experimental systems have been built in laboratories worldwide to investigate the influence of various environmental parameters on the efficacy of UV germicidal irradiation (UVGI) for deactivating airborne microorganisms. It is generally recognized that data from different laboratories might vary significantly due to differences in systems and experimental conditions. In this study we looked at the effect of the composition of the suspending medium on the size and UVGI susceptibility of Serratia marcescens in an experimental system built in our laboratory. S. marcescens was suspended in (i) distilled water, (ii) phosphate buffer, (iii) 10% fetal calf serum, (iv) phosphate-buffered saline (saline, 0.8% sodium chloride), and (v) synthetic saliva (phosphate-buffered saline with 10% fetal calf serum). At low humidity (36%), S. marcescens suspended in water-only medium was the most susceptible to UVGI, followed by those in serum-only medium. The count median diameters (CMDs) for culturable particles from water-only and serum-only media were 0.88 and 0.95 μm, respectively, with the measurements based on their aerodynamic behavior. The bacteria suspended in phosphate buffer, synthetic saliva, and phosphate-buffered saline had similar UVGI susceptibility and CMD at 1.0, 1.4, and 1.5 μm, respectively. At high humidity (68%) the CMD of the particles increased by 6 to 16%, and at the same time UVGI susceptibility decreased, with the magnitude of decrease related to the type of suspending medium. In conclusion, the choice of suspending medium influenced both size and UVGI susceptibility of S. marcescens. These data are valuable for making comparisons and deciding on the use of an appropriate medium for various applications.
PMCID: PMC383042  PMID: 15066792
Journal of Bacteriology  1962;84(6):1297-1302.
Zimmerman, Leonard (U.S. Army Biological Laboratories, Fort Detrick, Frederick, Md.). Survival of Serratia marcescens after freeze-drying or aerosolization at unfavorable humidity. I. Effects of sugars. J. Bacteriol. 84:1297–1302. 1962.—Suspensions of Serratia marcescens were subjected to freeze-drying or to aerosolization at unfavorable humidity levels. The survival of the cells during one or the other of these treatments was markedly improved in the presence of common sugars, but no one sugar stabilized the cells against both stresses. The protective effects of the sugars were correlated with their penetrability into cells; minimally penetrable sugars stabilized cells against aerosolization, and freely penetrable sugars stabilized cells during freeze-drying. These results were attributed to the modifications of intracellular water content induced by the presence of the sugars in the cell suspensions.
PMCID: PMC278062  PMID: 14003706
12.  New method for determination of efficacy of health care personnel hand wash products. 
Journal of Clinical Microbiology  1989;27(10):2295-2299.
A method of studying the effects of health care personnel hand wash products is described. The fingernail regions of the hands of volunteers are inoculated with a mixture of Escherichia coli and Serratia marcescens, and the areas are dried for a standard time. After routine hand washing, each fingernail region is individually scrubbed with an electric toothbrush which moves longitudinally to the handle into collection fluid contained in a petri dish. The test bacteria in the fluid are then enumerated. (Bacillus subtilis spores may be included as tracers to show degree of physical removal of the procedure.) This method has several advantages over the frequently used glove juice technique. Experimental designs with large numbers of volunteers, multiple sampling sites, and many hand wash products may be performed. Ten sampling sites (fingers) are available, versus the two gloved hands for testing products. (Efficiency is almost 100% in the recovery of spore tracers placed on the fingernails.) Many commercial health care personnel hand wash products containing antimicrobial agents substantive to the skin do not rapidly reduce numbers of inoculated bacteria in the fingernail regions to any greater extent than nonantimicrobial hand washes. Products containing isopropanol or ethanol are very effective in decreasing bacteria in areas around and under the fingernails.
PMCID: PMC267012  PMID: 2685028
13.  Microbiological Laboratory Hazard of Bearded Men 
Applied Microbiology  1967;15(4):899-906.
An investigation was conducted to evaluate the hypothesis that a bearded man subjects his family and friends to risk of infection if his beard is contaminated by infectious microorganisms while he is working in a microbiological laboratory. Bearded and unbearded men were tested with Serratia marcescens and Bacillus subtilis var. niger. Contact aerosol transmission from a contaminated beard on a mannequin to a suitable host was evaluated with both Newcastle disease virus and Clostridium botulinum toxin, type A. The experiments showed that beards retained microorganisms and toxin despite washing with soap and water. Although washing reduced the amount of virus or toxin, a sufficient amount remained to produce disease upon contact with a suitable host.
PMCID: PMC547091  PMID: 4963447
14.  Kinetic Analysis of Growth Rate, ATP, and Pigmentation Suggests an Energy-Spilling Function for the Pigment Prodigiosin of Serratia marcescens▿  
Journal of Bacteriology  2008;190(22):7453-7463.
Serratia marcescens is a gram-negative environmental bacterium and opportunistic pathogen. S. marcescens expresses prodigiosin, a bright red and cell-associated pigment which has no known biological function for producing cells. We present here a kinetic model relating cell, ATP, and prodigiosin concentration changes for S. marcescens during cultivation in batch culture. Cells were grown in a variety of complex broth media at temperatures which either promoted or essentially prevented pigmentation. High growth rates were accompanied by large decreases in cellular prodigiosin concentration; low growth rates were associated with rapid pigmentation. Prodigiosin was induced most strongly during limited growth as the population transitioned to stationary phase, suggesting a negative effect of this pigment on biomass production. Mathematically, the combined rate of formation of biomass and bioenergy (as ATP) was shown to be equivalent to the rate of prodigiosin production. Studies with cyanide inhibition of both oxidative phosphorylation and pigment production indicated that rates of biomass and net ATP synthesis were actually higher in the presence of cyanide, further suggesting a negative regulatory role for prodigiosin in cell and energy production under aerobic growth conditions. Considered in the context of the literature, these results suggest that prodigiosin reduces ATP production by a process termed energy spilling. This process may protect the cell by limiting production of reactive oxygen compounds. Other possible functions for prodigiosin as a mediator of cell death at population stationary phase are discussed.
PMCID: PMC2576671  PMID: 18805986
Journal of Bacteriology  1963;85(5):1136-1140.
Blizzard, John L. (University of Houston, Houston, Texas) and G. E. Peterson. Selective inhibition of proline-induced pigmentation in washed cells of Serratia marcescens. J. Bacteriol. 85:1136–1140. 1963.—Streptomycin, chloramphenicol, and tetracyclines inhibited the synthesis of prodigiosin by Serratia marcescens strain D1. This occurred at concentrations of the antibiotic too low to inhibit the growth of the organism in either agar media or broth cultures. Nonpigmented cells were produced in broth by either adding streptomycin or incubating at 37 C. After being washed and resuspended in aqueous saline containing either casein hydrolysate, l-proline, or a glycine-succinate mixture and incubated at 27 C for 24 hr, these cells formed pigment. The appearance of pigment was preceded by a lag period of 10 hr. Prodigiosin production by these washed suspensions of cells was completely inhibited by either streptomycin or glucose, or by incubation at 37 C instead of 27 C. Even though pigmentation by washed-cell suspensions was induced by proline, the utilization of proline was not affected by streptomycin or glucose, or by incubation at 37 C. To block pigmentation completely, streptomycin had to be added to proline-supplemented cells before they were 10 hr old. Addition of the antibiotic after the end of the induction period caused either partial or no inhibition of pigment production. Streptomycin caused an increase in the endogenous respiration of S. marcescens but failed to affect the constitutive enzymes that oxidize glucose. The possible relationships of these phenomena are discussed.
Weil (1952) reported that low concentrations of chloramphenicol and certain tetracyclines inhibit the synthesis of prodigiosin while permitting growth by Serratia marcescens. He noted the potential value to “mode-of-action” studies of an organism having certain functions selectively inhibited by antibiotics. We confirmed Weil's (1952) observations and found that streptomycin at low concentration would also inhibit the synthesis of prodigiosin without impeding growth.
Further studies of the selective inhibition of prodigiosin synthesis by streptomycin were performed using nonproliferating suspensions of washed cells (Gott and Williams, 1961). Either a glycine-succinate mixture or l-proline could cause nonproliferating cells to form pigment. A period of induction preceded the formation of pigment. Either streptomycin or glucose, or an incubation temperature of 37 C, inhibited the proline-induced pigmentation by washed cells. Further investigations provided insights to these findings.
PMCID: PMC278295  PMID: 14044006
16.  Preservation of Bacteria by Circulating-Gas Freeze Drying 
Applied Microbiology  1963;11(3):244-248.
Water-washed Serratia marcescens and Escherichia coli were freeze dried in a circulating-gas system at atmospheric pressure. This convective procedure resulted in a substantially higher survival of organisms than could be obtained by the vacuum method of freeze drying. There was little or no decrease in cell viability during convective drying when the residual moisture content was 15% or higher. Below this level, survival declined with decreasing moisture content. A detailed comparison of the convective and vacuum methods indicated that the advantage gained by freeze drying bacteria in air accrues in the early period of sublimation, at which time cells were found to be sensitive to vacuum drying but insensitive to air drying. An explanation for this difference is proposed, based upon the kinetics of water removal in the two processes. In brief, it is suggested that the convective method permits samples to be dried more uniformly; and regional over-drying, which may be deleterious even if transient, is thus avoided in achieving the optimal level of moisture.
PMCID: PMC1057981  PMID: 13998202
17.  Structure and function of a bacterial mRNA stabilizer: analysis of the 5' untranslated region of ompA mRNA. 
Journal of Bacteriology  1991;173(15):4578-4586.
The 5' untranslated region (UTR) of the Escherichia coli ompA transcript functions in vivo as a growth rate-regulated mRNA stabilizer. The secondary structure of this mRNA segment has been determined by a combination of three methods: phylogenetic analysis, in vitro probing with a structure-specific RNase, and methylation by dimethylsulfate in vivo and in vitro. These studies reveal that despite extensive sequence differences, the 5' UTRs of the ompA transcripts of E. coli, Serratia marcescens, and Enterobacter aerogenes can fold in a remarkably similar fashion. Furthermore, the Serratia and Enterobacter ompA 5' UTRs function as effective mRNA stabilizers in E. coli. Stabilization of mRNA by the Serratia ompA 5' UTR is growth rate dependent. These findings indicate that the features of the ompA 5' UTR responsible for its ability to stabilize mRNA in a growth rate-regulated manner are to be found among the structural similarities shared by these diverse evolutionary variants.
PMCID: PMC208132  PMID: 1713205
18.  Survival of Airborne Bacteria in a High Urban Concentration of Carbon Monoxide1 
Applied Microbiology  1973;25(1):86-91.
Vegetative cells of Serratia marcescens 8UK, Sarcina lutea, and spores of Bacillus subtilus var. niger were held in aerosols, with and without an urban concentration of CO (85 μliters per liter or ppm), for up to 6 hr at 15 C and a relative humidity (RH) of approximately 0, 25, 50, 75, and 95%. It was found that CO enhanced the death rate of S. marcescens 8UK at least four- to sevenfold at low RH (ca. 1 to 25%), but protected the cells at high RH (ca. 90%). Death rates of S. lutea, with or without added CO, were comparatively low over the entire RH range. However, in the first hour, airborne S. lutea held in CO-containing air were more stable than those in air without added CO (i.e., CO protection). A marked increase in the death rate (up to 70-fold) occurred in the subsequent 5 hr within the RH range of approximately 0 to 75%. Statistical analysis indicated that aerosol decay rates of B. subtilus var. niger spores decreased significantly, when held in a CO-containing as compared to a non-CO-containing atmosphere, in the 0 to 85% RH range. Thus, the data presented indicate that CO in the urban environment may have a protective or lethal effect on airborne bacteria, dependent upon at least the microbial species, aerosol age, and relative humidity. A mechanism for CO death enhancement and protection of airborne S. marcescens 8UK is suggested to involve CO uncoupling of an energy-requiring death mechanism and an energy-requiring maintenance mechanism at high and low RH, respectively.
PMCID: PMC380740  PMID: 4631439
19.  Sorption of Heterotrophic and Enteric Bacteria to Glass Surfaces in the Continuous Culture of River Water 
Applied Microbiology  1974;28(4):572-578.
A natural population of heterotrophic bacteria, including enterics, was observed to sorb to glass surfaces and multiply during the continuous culture of river water. An initial rate of attachment equivalent to a doubling time of about 2 h was observed with a corresponding increase in the suspended population. After 24 h both the sorbed and suspended populations stabilized with a mass doubling time approximating 100 h at a dilution rate of 0.012/h. On the basis of respiration and degradative enzymatic data, the sorbed microorganisms appeared to be somewhat more metabolically active than the organisms in suspension.
PMCID: PMC186774  PMID: 4424694
20.  R Factor-Mediated Antibiotic Resistance in Serratia marcescens 
Nineteen of 39 multiresistant strains of Serratia marcescens isolated from clinical sources transferred antibiotic resistance to Escherichia coli or Klebsiella pneumoniae recipients. Marcesins and/or phage prevented effective resistance transfer to E. coli and attempts to select marcescin-resistant mutants of the E. coli recipient strain were unsuccessful. Transfer of resistance was demonstrated for all drugs tested except nalidixic acid. Approximately 90% of donors resistant to tobramycin, ampicillin, or carbenicillin transferred resistance to these drugs. High levels of transferred resistance (minimal inhibitory concentration, >2,500 μg/ml) were demonstrated particularly for ampicillin, carbenicillin, and kanamycin. Transmissibility of Serratia R factors was greatest between isogeneic strains of E. coli K-12. Comparative rates of spontaneous loss of R factor-mediated resistance indicated that Serratia R factors are less stable in E. coli and K. pneumoniae transcipients than in the indigenous hosts.
PMCID: PMC429700  PMID: 791085
21.  Prolonged survival of Serratia marcescens in chlorhexidine. 
During an outbreak of Serratia marcescens infections at our hospital, we discovered widespread contamination of the 2% chlorhexidine hand-washing solution by S. marcescens. Examination by electron microscopy of the sides of bottles in which this solution was stored revealed that microorganisms were embedded in a fibrous matrix. Bacteria, free in the liquid, were morphologically abnormal, showing cell wall disruption or cytoplasmic changes. Furthermore, bacteria adherent to the walls of the storage jugs and embedded in this fibrous matrix also had morphologically abnormal cytoplasm. Despite these changes, viable S. marcescens organisms were recovered from the fluid during a storage period of 27 months. The concentration of chlorhexidine required to inhibit these strains of Serratia was 1,024 microgram/ml; however, the organism could survive in concentrations of up to 20,000 micrograms/ml. Additional studies are needed to define the mechanism(s) that allows such bacteria to contaminate and survive in disinfectants.
PMCID: PMC244159  PMID: 7032422
22.  Adaptation and growth of Serratia marcescens in contact lens disinfectant solutions containing chlorhexidine gluconate. 
Serratia marcescens (11 of 12 strains) demonstrated an ability to grow in certain chlorhexidine-based disinfecting solutions recommended for rigid gas-permeable contact lenses. For a representative strain, cells that were grown in nutrient-rich medium, washed, and inoculated into disinfecting solution went into a nonrecoverable phase within 24 h. However, after 4 days, cells that had the ability to grow in the disinfectant (doubling time, g = 5.7 h) emerged. Solutions supporting growth of S. marcescens were filter sterilized. These solutions, even after removal of the cells, showed bactericidal activity against Pseudomonas aeruginosa and a biphasic survival curve when rechallenged with S. marcescens. Adaptation to chlorhexidine by S. marcescens was not observed in solutions formulated with borate ions. For chlorhexidine-adapted cells, the MIC of chlorhexidine in saline was eightfold higher than that for unadapted cells. Cells adapted to chlorhexidine showed alterations in the proteins of the outer membrane and increased adherence to polyethylene. Cells adapted to chlorhexidine persisted or grew in several other contact lens solutions with different antimicrobial agents, including benzalkonium chloride.
PMCID: PMC202075  PMID: 8439148
23.  Prodigiosin-Producing Bacteria from Marine Sources1 
Applied Microbiology  1964;12(1):13-17.
Two aerobic, gramnegative, red-pigmented, rod-shaped bacteria were compared morphologically and physiologically with Serratia species, which they resembled superficially. The pigment produced by the marine isolates was shown to be similar to prodigiosin, the red pigment of S. marcescens. The isolates had a single polar flagellum, were oxidative, and did not produce acetoin from glucose or reduce nitrates, which made them distinct from both S. marcescens and S. marinorubra. The latter conformed well to the descriptions of S. marcescens in Bergey's Manual. The marine isolates displayed an absolute growth requirement for sea water or its equivalent. The growth requirement for sea water was replaced by sea-water levels of sodium, potassium, and magnesium chloride. Pigment was produced only when this salt mixture was further supplemented with calcium chloride. Neither sea water nor a high salt level was required for growth or prodigiosin synthesis by the Serratia species examined.
PMCID: PMC1058056  PMID: 14106932
24.  Induction of Pigmentation in Nonproliferating Cells of Serratia marcescens by Addition of Single Amino Acids 
Journal of Bacteriology  1971;106(2):444-448.
Addition of casein hydrolysate to suspensions of washed, nonpigmented, nonproliferating Serratia marcescens incubating at 27 C induced biosynthesis of prodigiosin. Four amino acids of casein hydrolysate, dl-aspartic acid, l-glutamic acid, l-proline, and l-alanine caused formation of pigment when added individually. dl-Ornithine also was effective. Optimal concentrations for maximal pigmentation were 5 to 10 mg/ml; at these high concentrations, d-serine also induced biosynthesis of some prodigiosin. dl-Alanine and -ornithine were as effective as the l-iosomers, but l-glutamic acid and l-proline gave better responses than their racemic mixtures. Kinetics of prodigiosin biosynthesis after addition of dl-alanine (20 mg/ml) were similar to those of cells suspended in 0.2% casein hydrolysate. The other amino acids were less effective. Addition of 5 mg of dl-alanine or casein hydrolysate per ml to minimal medium increased by 30% the amount of prodigiosin formed by growing cells after incubation for 7 days at 27 C. Cultures grown for 7 days at 27 C in 0.2% casein hydrolsate formed more prodigiosin than did suspensions of nonproliferating cells containing individual amino acids or casein hydrolysate. However, more pigment was produced by cells suspended in l-alanine (5 mg/ml) or l-proline (10 mg/ml) than when suspended in 0.4% natural or synthetic casein hydrolysate. Filtrates from suspensions of nonproliferating cells forming pigment in l-proline induced more rapid formation of prodigiosin, but filtrates from suspensions in dl-alanine did not. The data supported the hypothesis that pyrrole groups of prodigiosin may be synthesized from 5-carbon amino acids such as proline, ornithine, aspartic, and glutamic acids, but the role of alanine is unknown.
PMCID: PMC285115  PMID: 4929860
25.  A novel medium for the enhanced cell growth and production of prodigiosin from Serratia marcescens isolated from soil 
BMC Microbiology  2004;4:11.
Prodigiosin produced by Serratia marcescens is a promising drug owing to its reported characteristics of having antifungal, immunosuppressive and antiproliferative activity. From an industrial point of view the necessity to obtain a suitable medium to simultaneously enhance the growth of Serratia marcescens and the pigment production was the aim of this work. The usage of individual fatty acid as substrate in industries would be cost-effective in the long run and this paved the way for us to try the effect of different fatty acid-containing seeds and oils of peanut, sesame and coconut as source of substrate.
The addition of sugars only showed slight enhancement of prodigiosin production in nutrient broth but not in fatty acid containing seed medium. The powdered peanut broth had supported better growth of Serratia marcescens and higher yield of prodigiosin when compared with the existing nutrient broth and peptone glycerol broth. A block in prodigiosin production was seen above 30°C in nutrient broth, but the fatty acid seed medium used by us supported prodigiosin production upto 42°C though the yields were lower than what was obtained at 28°C. From the results, the fatty acid form of carbon source has a role to play in enhanced cell growth and prodigiosin production.
We conclude by reporting that the powdered and sieved peanut seed of different quality grades were consistent in yielding a fourty fold increase in prodigiosin production over the existing media. A literature survey on the composition of the different media components in nutrient broth, peptone glycerol broth and the fatty acid containing seeds and oils enabled us to propose that the saturated form of fatty acid has a role to play in enhanced cell growth and prodigiosin production. This work has also enabled us to report that the temperature related block of prodigiosin biosynthesis varies with different media and the powdered peanut broth supports prodigiosin production at higher temperatures. The medium suggested in this work is best suitable from an industrial point of view in being economically feasible, in terms of the higher prodigiosin yield and the extraction of prodigiosin described in this paper is simple with minimal wastage.
PMCID: PMC404375  PMID: 15113456

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