Loss Of Thermotolerance Research Articles

Loss of thermotolerance in antibiotic-resistant Acinetobacter baumannii

ABSTRACT Bacterium Acinetobacter baumannii is a leading cause of nosocomial infections. The occurrence of antibiotic-resistant A. baumannii isolates outside hospitals suggests that monitoring of this pathogen in environmental samples is needed. Survival of pandrug-resistant A. baumannii was followed on selective plates with and without carbapenems in water and soil. After a few days of starvation, A. baumannii lost the ability to be cultivated at 44°C on plates supplemented with carbapenems. Once cultivated on plates without carbapenems and/or at 36°C, A. baumannii could grow again at 44°C in the presence of carbapenems. Comparative proteomic analysis revealed that impaired membrane integrity and reduced function of efflux pumps due to elevated temperature combined with antibiotic exposure were the main reasons for this phenomenon. Loss of thermotolerance in the presence of antibiotics points to the need for temperature adjustment in long-term monitoring of A. baumannii in environmental samples, to avoid the underestimation of viable bacteria.

Reduction of Optimal Thermal Range in Aging Western Cherry Fruit Flies (Diptera: Tephritidae).

The western cherry fruit fly is an economically important pest of sweet cherries in the western United States. The potential of this pest to establish and spread in areas in which it is not currently present has been the focus of recent research. Most published information on the thermal tolerance and optimal thermal range of this pest has focused primarily on the diapausing pupae and predictive phenology models. Microrespirometry and differential calorimetry can be useful tools in describing the thermotolerance and optimal thermal range of insects. This methodology was employed to investigate the effects of western cherry fruit fly adult age on the optimal thermal range. Newly emerged flies exhibited the widest optimal thermal range spanning from 6.6 to 42.2°C for a total range of 35.8°C during heating scans of 0.4°C/min from 2 to 50°C. This range diminished as the flies aged, with the shortest span observed with 28-d-old flies ranging from 10.5 to 37.8°C, a span of 27.2°C. Measurements of heat rate and oxygen consumption at isothermal, or static, temperatures indicated that all flies could survive exposure to 40°C for at least 20 min, and that metabolism was greatly reduced, with a concomitant reduction in oxygen consumption rate at 40 to 42°C. All flies exhibited a heat rate and oxygen consumption rate of zero when exposed to 45 and 50°C. The loss of thermotolerance in adult flies can influence its ability to establish and spread in climates where daily temperatures exceed the optimal thermal range of this species.

Heat Inactivation of Escherichia coli O157:H7 in Apple Juice Exposed to Chlorine

Exposure of Escherichia coli O157:H7 to chlorine before heat treatment results in increased production of heat shock proteins. Current heating regimens for pasteurizing apple cider do not account for chlorine exposure in the wash water. This research determined the effect of sublethal chlorine treatment on thermal inactivation of E. coli O157:H7. D58-values were calculated for stationary-phase cells exposed to 0.6 mg/liter of total available chlorine and unchlorinated cells in commercial shelf-stable apple juice (pH 3.6). D58-values for unchlorinated and chlorine-exposed cells in buffer were 5.45 and 1.65 min, respectively (P < 0.01). Death curves of chlorine-exposed and unchlorinated cells in apple juice were not completely linear. Unchlorinated cells heated in apple juice exhibit a 3-min delay before onset of linear inactivation. Chlorine treatment eliminated this shoulder, indicating an overall loss of thermotolerance. The linear portion of each curve represented a small fraction of the total population. D58-values calculated from these populations are 0.77 min for unexposed cells and 1.19 min for chlorine-exposed cells (P = 0.05). This indicates that a subpopulation of chorine-treated cells is possibly more resistant to heat because of chlorine treatment. The effect of chlorine treatment, however, is insignificant when compared with the effect of losing the shoulder. This is illustrated by the time required to kill the initial 90% of the cell population. This is observed to be 3.14 min for unchlorinated versus 0.3 min for chlorine-exposed cells (P < 0.001). These observations indicate that current heat treatments need not be adjusted for the effect of chlorine treatment.

Disruption of the HSF3 gene results in the severe reduction of heat shock gene expression and loss of thermotolerance.

The vertebrate genome encodes a family of heat shock factors (HSFs 1-4) of which the DNA-binding and transcriptional activities of HSF1 and HSF3 are activated upon heat shock. HSF1 has the properties of a classical HSF and exhibits rapid activation of DNA-binding and transcriptional activity upon exposure to conditions of heat shock and other stresses, whereas HSF3 typically is activated at higher temperatures and with distinct delayed kinetics. To address the role of HSF3 in the heat shock response, null cells lacking the HSF3 gene were constructed by disruption of the resident gene by somatic recombination in an avian lymphoid cell line. Null cells lacking HSF3, yet expressing normal levels of HSF1, exhibited a severe reduction in the heat shock response, as measured by inducible expression of heat shock genes, and did not exhibit thermotolerance. At intermediate heat shock temperatures, where HSF1 oligomerizes to an active trimer in wild-type cells, HSF1 remained as an inert monomer in the HSF3 null cell line. HSF3 null cells were restored to a nearly normal heat shock-responsive state by reintroduction of an exogenous HSF3 gene. These results reveal that HSF3 has a dominant role in the regulation of the heat shock response and directly influences HSF1 activity.

Induction of Hsp72 and transient nuclear localization of Hsp73 and Hsp72 correlate with the acquisition and loss of thermotolerance in postimplantation rat embryos

A number of cell culture studies have indicated that there is a positive correlation between the induction and decay of thermotolerance and the kinetics of Hsp72 expression. In this study, we have demonstrated that, in gestational day 10 embryos, induction and decay of thermotolerance occur over an 8 hr period. To test the hypothesis that there is a correlation between loss of thermotolerance and the decline of Hsp72 or Hsp73 gene products over time, expression levels of both Hsp72 and constitutively expressed Hsp73 mRNAs and proteins were examined at several time points following exposure to a thermotolerance-inducing exposure of 42°C. Our results indicated that Hsp72 mRNA was strongly induced 1 hr after exposure but no longer detectable by 8 hr. Although our Western blot results indicated that Hsp72 protein was present beyond 8 hr after exposure, Northern blot analysis showed that Hsp72 mRNA was no longer present 5 hr after exposure to 42°C. The latter finding indicates that no new Hsp72 can be synthesized at this time point and beyond. Although there was very little or no induction of Hsp73, immunohistochemical analysis revealed a dramatic, transient shift in intracellular localization of Hsp73 protein, as well as Hsp72. Under non-stress conditions, Hsp73 was cytoplasmically localized but localization was largely nuclear 1 hr after exposure, when thermotolerance was demonstrable. Hsp73 and Hsp72 proteins were no longer localized in the nucleus by 8 hr, when thermotolerance was no longer detectable. Thus, the induction of Hsp72 and the transient nuclear localization of both Hsp72 and Hsp73 correlate with the kinetics of thermotolerance in the postimplantation rat embryo. Dev. Dyn. 208:227–243, 1997. © 1997 Wiley-Liss, Inc.

Induction of Hsp72 and transient nuclear localization of Hsp73 and Hsp72 correlate with the acquisition and loss of thermotolerance in postimplantation rat embryos.

A number of cell culture studies have indicated that there is a positive correlation between the induction and decay of thermotolerance and the kinetics of Hsp72 expression. In this study, we have demonstrated that, in gestational day 10 embryos, induction and decay of thermotolerance occur over an 8 hr period. To test the hypothesis that there is a correlation between loss of thermotolerance and the decline of Hsp72 or Hsp73 gene products over time, expression levels of both Hsp72 and constitutively expressed Hsp73 mRNAs and proteins were examined at several time points following exposure to a thermotolerance-inducing exposure of 42 degrees C. Our results indicated that Hsp72 mRNA was strongly induced 1 hr after exposure but no longer detectable by 8 hr. Although our Western blot results indicated that Hsp72 protein was present beyond 8 hr after exposure, Northern blot analysis showed that Hsp72 mRNA was no longer present 5 hr after exposure to 42 degrees C. The latter finding indicates that no new Hsp72 can be synthesized at this time point and beyond. Although there was very little or no induction of Hsp73, immunohistochemical analysis revealed a dramatic, transient shift in intracellular localization of Hsp73 protein, as well as Hsp72. Under non-stress conditions, Hsp73 was cytoplasmically localized but localization was largely nuclear 1 hr after exposure, when thermotolerance was demonstrable. Hsp73 and Hsp72 proteins were no longer localized in the nucleus by 8 hr, when thermotolerance was no longer detectable. Thus, the induction of Hsp72 and the transient nuclear localization of both Hsp72 and Hsp73 correlate with the kinetics of thermotolerance in the postimplantation rat embryo.

Effect of pre- and post-heat shock temperature on the persistence of thermotolerance and heat shock-induced proteins in Listeria monocytogenes.

The effect of incubation temperature, before and after a heat shock, on thermotolerance of Listeria monocytogenes at 58 degrees C was investigated. Exposing cells grown at 10 degrees C and 30 degrees C to a heat shock resulted in similar rises in thermotolerance while the increase was significantly higher when cells were grown at 4 degrees C prior to the heat shock. Cells held at 4 degrees C and 10 degrees C after heat shock maintained heat shock-induced thermotolerance for longer than cells held at 30 degrees C. The growth temperature prior to inactivation had negligible effect on the persistence of heat shock-induced thermotolerance. Concurrent with measurements of thermotolerance were measurements of the levels of heat shock-induced proteins. Major proteins showing increased synthesis upon the heat shock had approximate molecular weights of 84, 74, 63, 25 and 19 kDa. There was little correlation between the loss of thermotolerance after the heat shock and the levels of these proteins. Thermotolerance of heat shocked and non-heat shocked cells was described by traditional log-linear kinetics and a model describing a sigmoidal death curve (logistic model). Employing log-linear kinetics resulted in a poor fit to a major part of the data whereas a good fit was achieved by the use of a logistic model.

Selection of mutant Streptomyces strains with enhanced ability for acid precipitable polymeric lignin production

AbstractStreptomyces viridosporus T7A spores were subjected to ultraviolet (UV) light mutagensis. Two mutant strains S. viridosporus T7A‐40 and T7A‐198 were isolated. These strains were grown on 5 g of sugar cane baggase lignocellulose and produced 36.0 mg and 41.4 mg of acid precipitable polymeric lignin (APPL) respectively as compared to 25.1 mg produced by the wild type after 8 weeks incubation at 37 °C. The quantities of APPL produced by the mutants represent 43.7% and 65.3% more APPLs than those produced by the wild type. Apart from the increase in APPL production, the observable effects of the ultraviolet light included; variation in spore pigmentation, induction of a brown diffusible pigment, production of hydrogen sulfide, loss of thermotolerance, lower cellulase activity and loss of inositol utilizing ability.