Preincubation Temperature Research Articles

Differential temperature sensitivity of intracellular metabolic processes and extracellular soil enzyme activities

Abstract. Predictions concerning the feedback of soil heterotrophic respiration to a warming climate often do not differentiate between the extracellular and intracellular steps involved in soil organic matter decomposition. This study examined the temperature sensitivities of intracellular metabolic processes and extracellular soil enzyme activities and how they are influenced by previous temperatures. We pre-incubated soils at 5, 15, or 26 ∘C to acclimatize the microbial communities to different thermal regimes for 60 d before measuring potential activities of β-glucosidase and chitinase (extracellular enzymes), glucose-induced respiration (intracellular metabolic processes), and basal respiration at a range of assay temperatures (5, 15, 26, 37, and 45 ∘C). A higher pre-incubation temperature decreased the soil pH and C/N ratio and decreased β-glucosidase potential activity and respiration but not chitinase potential activity. It is likely that this legacy effect on β-glucosidase and respiration is an indirect effect of substrate depletion rather than physiological acclimatation or genetic adaptation. Pre-incubation temperature effects on temperature sensitivity were subtle and restricted to extracellular activities, perhaps because of the short (60 d) duration of the pre-incubation at temperatures that were below the initial optimum (∼ 30 ∘C) for the mesophilic soil community. However, we found that the intracellular and extracellular steps differ in their temperature sensitivity, and this observation differs depending on the range of temperature used for Q10 estimates of temperature sensitivity. Between 5 and 15 ∘C intracellular and extracellular processes show equal temperature sensitivity, but between 15 and 26 ∘C intracellular metabolic processes were more temperature sensitive than extracellular enzyme activity, and between 26 and 37 ∘C extracellular enzyme activity was more temperature sensitive than intracellular metabolic processes. This result implies that depolymerization of higher molecular weight carbon is more sensitive to temperature changes at higher temperatures (e.g. higher temperatures on extremely warm days), but the respiration of the generated monomers is more sensitive to temperature changes at moderate temperatures (e.g. mean daily maximum soil temperature). However, studies using multiple soil types and a greater range of pre-incubation temperatures are required to generalize our results. Nevertheless, since climate change predictions currently indicate that there will be a greater frequency and severity of hot summers and heatwaves, it is possible that global warming may reduce the importance of extracellular depolymerization relative to intracellular metabolic processes as the rate-limiting step of soil organic matter mineralization. We conclude that extracellular and intracellular steps are not equally sensitive to changes in soil temperature and that the previous temperature a soil is exposed to may influence the potential activity, but not temperature sensitivity, of extracellular and intracellular processes.

Normal Thermostability of p.Ser113Leu and p.Arg631Cys Variants of Mitochondrial Carnitine Palmitoyltransferase II (CPT II) in Human Muscle Homogenate.

Previous fibroblast and recombinant enzyme studies showed a markedly thermolabile p.Ser113Leu variant compared to the wild-type (WT) in muscle carnitine palmitoyltransferase II (CPT II) deficiency. Additionally, it has been shown that cardiolipin (CLP) stimulated or inhibited the p.Ser113Leu recombinant variant depending on the pre-incubation temperatures. In this study, the thermolabilities of mitochondrial enzyme CPT II in muscle homogenates of patients with the p.Ser113Leu (n = 3) and p.Arg631Cys (n = 2) variants were identified to be similar to that of WT. Pre-incubation with CLP on ice stimulated the WT enzyme more than both variants. However, CLP stimulated the variants and WT at 46 °C to about 6-18-fold. The present data indicate that the thermostability of CPT II variant in muscle homogenate is similar to that of WT. This is in contrast to the increased thermolability of enzymes derived from fibroblast and that of recombinant enzymes. Hence, it can be speculated that the disruption of the compartmentation in muscle homogenate mediates a protective effect on the thermolability of the native variant. However, the exact mechanism remains unclear. However, the activating effect of CLP on CPT II in muscle homogenate seems to align with those on recombinant enzymes.

Priming effect of stable C pool in soil and its temperature sensitivity

It is important to understand the effects of exogenous carbon (C) additions on the mineralization of native soil C in agricultural soils under changing global temperatures. The slowly mineralizable C pools represent the major component of soil carbon. Once the labile fraction of soil C is depleted through mineralization, additions of exogenous labile C may enhance decomposition of slowly mineralizable soil C through the so-called priming effect (PE). The effect of exogenous C on mineralization of slowly mineralizable C in soils that have undergone extended substrate depletion is not well understood. The aim of this study was to assess the effects of exogenous C addition on the PE and its temperature sensitivity (Q10) in two substrate depleted soils. The Mollisols with two levels of C 6.8% and 2.9% were pre-incubated at 15 and 25 ℃ for 864 days, followed by 42-day incubations with and without 13C-enriched corn (Zea mays) leaf addition at 15 and 25 ℃. Soil organic carbon (SOC) decreased by 7%−9% in low and 1%−3% in high C soil after the long-term pre-incubation, respectively. In the 42-day incubation following the long-term pre-incubation, the incubation at the higher temperature (25 ℃) significantly increased cumulative CO2 production compared to that at the lower temperature (15 °C) for both high and low C soils with or without exogenous C addition. Furthermore, corn leaves addition stimulated CO2 production derived from resistant OC in both soils, resulting in a positive PE. Higher incubation temperature resulted in a greater PE in both soils independent of pre-incubation temperature. The temperature sensitivity (Q10) of the PE in the low C soil pre-incubated at 25 °C was greater than that pre-incubated at 15 °C, but in the high C soil, the Q10 of PE was lower after pre-incubation at 25 °C. The results confirmed that exogenous C addition and a higher temperature accelerate native SOC mineralization and suggested that the temperature sensitivity of the PE depended on the soil C content and the long-term temperature regime a soil is subjected to.

Characterization of Neoagarooligosaccharide Hydrolase BpGH117 from a Human Gut Bacterium Bacteroides plebeius.

α-Neoagarobiose (NAB)/neoagarooligosaccharide (NAO) hydrolase plays an important role as an exo-acting 3,6-anhydro-α-(1,3)-L-galactosidase in agarose utilization. Agarose is an abundant polysaccharide found in red seaweeds, comprising 3,6-anhydro-L-galactose (AHG) and D-galactose residues. Unlike agarose degradation, which has been reported in marine microbes, recent metagenomic analysis of Bacteroides plebeius, a human gut bacterium, revealed the presence of genes encoding enzymes involved in agarose degradation, including α-NAB/NAO hydrolase. Among the agarolytic enzymes, BpGH117 has been partially characterized. Here, we characterized the exo-acting α-NAB/NAO hydrolase BpGH117, originating from B. plebeius. The optimal temperature and pH for His-tagged BpGH117 activity were 35 °C and 9.0, respectively, indicative of its unique origin. His-tagged BpGH117 was thermostable up to 35 °C, and the enzyme activity was maintained at 80% of the initial activity at a pre-incubation temperature of 40 °C for 120 min. Km and Vmax values for NAB were 30.22 mM and 54.84 U/mg, respectively, and kcat/Km was 2.65 s−1 mM−1. These results suggest that His-tagged BpGH117 can be used for producing bioactive products such as AHG and agarotriose from agarose efficiently.

Processing of Flavor-Enhanced Oils: Optimization and Validation of Multiple Headspace Solid-Phase Microextraction-Arrow to Quantify Pyrazines in the Oils.

An efficient and effective multiple headspace-solid phase microextraction-arrow-gas chromatography-mass spectrometry (MHS-SPME-arrow-GCMS) analytical protocol is established and used to quantify the flavor compounds in oils. SPME conditions, such as fiber coating, pre-incubation temperature, extraction temperature, and time were studied. The feasibility was compared between SPME-arrow and the traditional fiber by loading different sample amounts. It was found that the SPME-arrow was more suitable for the MHS-SPME. The limit of detection (LODs) and limit of quantitation (LOQs) of pyrazines were in the range of 2–60 ng and 6–180 ng/g oil, respectively. The relative standard deviation (RSD) of both intra- and inter-day were lower than 16%. The mean recoveries for spiked pyrazines in rapeseed oil were in the range of 91.6–109.2%. Furthermore, this newly established method of MHS-SPME-arrow was compared with stable isotopes dilution analysis (SIDA) by using [2H6]-2-methyl-pyrazine. The results are comparable and indicate this method can be used for edible oil flavor analysis.

Comparative analysis of unwashed and single washed mince gel from Indian major carps.

The gelling properties and quality characteristics of unwashed and single washed mince of catla, rohu and mrigal have been investigated to find out suitability of Indian major carps for the preparation of mince gel. The higher moisture content and lower protein content was reported in the single washed mince. The single washing of mince did not improve the gel strength. The gel strength showed significant difference (p < 0.05) and decreased in single washed mince than its unwashed counterparts in catla and mrigal except rohu. It has been observed that gel did not set at pre-incubation temperature of 40°C for 30min treatment. SDS-PAGE patterns of proteins did not show any loss of myosin heavy chain (MHC) in single washed mince of Indian major carps. Texture profile analysis showed higher hardness in washed mince gel of Indian major carps while, non-significant difference (p > 0.05) was observed in cohesiveness, adhesiveness and elasticity properties. The whiteness index of washed mince showed improvement. The overall study indicated that mince gels can be made from unwashed mince of Indian major carps, alleviating the problems of waste water disposal leading to production of more value added products with better nutritional value.

Protein Stability and Functional Characterization of Intra-Melanosomal Domain of Human Recombinant Tyrosinase-Related Protein 1.

Pigmentation is the result of a complex process by which the biopolymer melanin is synthesized and packed into melanosomes of melanocytes. Various types of oculocutaneous albinism (OCA), a series of autosomal recessive disorders, are associated with reduced pigmentation in the skin, eyes, and hair due to genetic mutations of proteins involved in melanogenesis. Human tyrosinase (Tyr) and tyrosinase-related protein 1 (Tyrp1) drives the enzymatic process of pigment bio-polymerization. However, within the melanogenic pathway, Tyrp1 has catalytic functions not clearly defined and distinct from Tyr. Here, we characterize the biochemical and biophysical properties of recombinant human Tyrp1. For this purpose, we purified and analyzed the intra-melanosomal domain (Tyrp1tr) for protein stability and enzymatic function in conditions mimicking the environment within melanosomes and the endoplasmic reticulum. The study suggests that Tyrp1tr is a monomeric molecule at ambient temperatures and below (<25 °C). At higher temperatures, >31 °C, higher protein aggregates form with a concurrent decrease of monomers in solution. Also, Tyrp1tr diphenol oxidase activity at pH 5.5 rises as both the pre-incubation temperature and the higher molecular weight protein aggregates formation increases. The enhanced protein activity is consistent with the volume exclusion change caused by protein aggregates.

Effects of preincubation on the gelatinization of cassava and corn starch suspensions containing sodium hydroxide as a main component of corrugating adhesives

Effects of the preincubation temperature and the caustic-ratio, the molar ratio of sodium hydroxide to starch glucose residue, on the gelatinization of cassava starch and corn starch suspensions were studied using differential scanning calorimetry in view of utilization for corrugating adhesives. The gelatinization temperature and enthalpy change of cassava starch suspensions after the preincubation at 30°C decreased as the caustic-ratio increased, similar to those of corn starch ones: The gelatinization starting temperature (Ts) decreased considerably more than the peak temperature and the conclusion temperature (Tc). Although Ts lowered and the width of gelatinization temperature expanded, compared with those of corn starch suspensions, the two starch suspensions with the same half gelatinization transition temperature showed similar gelatinization characteristics of almost the same Ts and Tc. During 1 h-preincubations at 30°C–50°C, the starch granules with Ts that were lowered considerably below each preincubation temperature by sodium hydroxide showed limited gelatinization. The gelatinization transition did not rapidly spread over the whole suspension, but progressed stepwise in response to the increase of the causticratio and the rise of the preincubation temperature. In a prolonged preincubation at a constant temperature, Ts gradually rose at higher caustic-ratios in which stepwise gelatinization commenced. Although the starch gelatinization was irreversible and not in a stable equilibrium state for a long time, we concluded that such stepwise gelatinization progress controlled the practical preparation and use of corrugating adhesives.

Determination of optimal conditions for activity of arginase in kidney of rabbit

Optimum conditions for arginase in kidney tissue may differ from those for arginase of liver. Because optimum conditions for measuring arginase activity in rabbit kidney tissue were not determined, it was aimed to optimize arginase of rabbit kidney tissue and compare it with arginase of liver. In the present study, the kidneys of six New Zealand rabbits aged 1 year old were used. It was found to be preincubation temperature 53 °C, preincubation period is 15 minutes, incubation period is 10 minutes and optimum pH is 10.1 in NaHCO3-NaOH buffer for arginase in rabbit kidney tissue. Enzyme achieved its highest activity at 1 mM MnCl2concentration. Km of arginase in rabbit kidney tissue for L-arginine was measured as 12.5 mM. It was determined that Mn+2 ions and preincubation at 53 °C were essential for the activation of the enzyme. As a result, optimal conditions for the kidney arginase were found to be different from those for the liver arginase. Our study indicates that arginases encoded by separate gene loci may have different optimal conditions

Altered embryonic development in northern bobwhite quail (Colinus virginianus) induced by pre-incubation oscillatory thermal stresses mimicking global warming predictions.

Global warming is likely to alter reproductive success of ground-nesting birds that lay eggs normally left unattended for days or even weeks before actual parental incubation, especially in already warm climates. The native North American bobwhite quail (Colinus virginianus) is such a species, and pre-incubation quail eggs may experience temperatures ≥45°C. Yet, almost nothing is known about embryonic survival after such high pre-incubation temperatures. Freshly laid bobwhite quail eggs were exposed during a 12 day pre-incubation period to one of five thermal regimes: low oscillating temperatures (25–40°C, mean = 28.9°C), high oscillating temperatures (30–45°C, mean = 33.9°C), low constant temperatures (28.85°C), high constant temperatures (mean = 33.9°C), or commercially employed pre-incubation temperatures (20°C). After treatment, eggs were then incubated at a standard 37.5°C to determine subsequent effects on embryonic development rate, survival, water loss, hatching, and embryonic oxygen consumption. Both quantity of heating degree hours during pre-incubation and specific thermal regime (oscillating vs. non-oscillating) profoundly affected important aspects of embryo survival and indices of development and growth Pre-incubation quail eggs showed a remarkable tolerance to constant high temperatures (up to 45°C), surviving for 4.5±0.3 days of subsequent incubation, but high oscillating pre-incubation temperature increased embryo survival (mean survival 12.2±1.8 days) and led to more rapid development than high constant temperature (maximum 38.5°C), even though both groups experienced the same total heating degree-hours. Oxygen consumption was ~200–300 μl O2.egg.min-1 at hatching in all groups, and was not affected by pre-incubation conditions. Oscillating temperatures, which are the norm for pre-incubation quail eggs in their natural habitat, thus enhanced survival at higher temperatures. However, a 5°C increase in pre-incubation temperature, which equates to the predicted long-term increases of 5°C or more, nonetheless reduced hatching rate by approximately 50%. Thus, while pre-incubation bobwhite eggs may be resiliant to moderate oscillating temperature increases, global warming will likely severely impact wild bobwhite quail populations, especially in their strongholds in southern latitudes.

Thermal treatments prior to and during the beginning of incubation affects development of the broiler embryo and yolk sac membranes, and live performance and carcass characteristics

This study evaluated temperature during preincubation and embryonic day 0 (E0) E0 to E5 of incubation on broiler embryo development and subsequent live performance. Freshly laid eggs from a single 41-wk-old Ross 708 broiler breeder flock produced on a single day were weighed individually for weight matching purposes, stored overnight, and assigned to 4 treatment combinations of 2 preincubation temperatures (23.9 or 29.4°C)× 2 E0 to E5 temperatures (38.1 or 37.5°C). The 29.4°C preincubation temperature decreased (P ≤ 0.05) yolk sac membrane (YSM) vasculature at E6 and E7, and increased (P ≤ 0.05) embryo weight and length but decreased (P ≤ 0.05) yolk sac weight (YSW) at E15. No subsequent main effects were observed. The 38.1°C incubation temperature increased YSM vasculature at E7, chorioallantoic membrane (CAM) vasculature at E8 and E10, and egg weight loss, embryo weight, and embryo length at E15 and chick length at E21 in the presence of reduced BW and YSW (P ≤ 0.05). This was followed by greater male BW at 35 d, as well as improved FCR in females 0 to 14 d and in males 15 to 35 d (P ≤ 0.05). Pectoralis major and minor yields were increased (P ≤ 0.05) at 50 d of age in males and females, respectively. There were no interactions observed with regards to broiler live performance and carcass yield, which probably negated the importance of the interactions observed for preincubation temperature by E0 to E5 incubation temperature that affected YSM vasculature, CAM vasculature area, egg weight loss, embryo weight, yolk sac weight, and chick length.

Thermal acclimation of organic matter decomposition in an artificial forest soil is related to shifts in microbial community structure

Thermal acclimation of soil organic matter (SOM) decomposition is frequently observed and has often been attributed to substrate depletion under warming, but other mechanisms, such as changes in microbial community structure and functioning, have received less attention. In order to determine whether shifts in microbial community structure and functioning are involved in thermal acclimation of SOM decomposition, a laboratory incubation experiment was conducted using an artificial forest soil. Samples were first subjected to different temperatures of 5, 15, and 25 °C during a 72-day pre-incubation period and then half of the microcosms from each pre-incubation temperature were incubated at 5 or 25 °C for a period of 11 days. Substantial thermal acclimation of SOM decomposition was observed, with the SOM decomposition in soils pre-incubated at higher temperatures being less sensitive to temperature. Along with the reduced temperature sensitivity in response to warming, significant changes in microbial community PLFAs, microbial biomass carbon (MBC), and the potential activities of 11 enzymes were also observed. Nevertheless, shifts in microbial community PLFAs and particular enzyme activities provided the most explanatory power for the decreased temperature sensitivity with warming, as revealed by a multivariate regression analysis. The microbial community structure shifts were mainly manifested as an increase in the relative abundance of Gram-positive bacteria and decreases in the relative abundances of Gram-negative bacteria and fungi. Microbial communities pre-incubated under lower temperatures experienced greater shifts in their structure. Substrate depletion did not occur in this short-term incubation experiment, since neither total organic carbon (TOC) nor dissolved organic carbon (DOC) decreased with increasing temperature. Our results suggest that shifts in microbial community structure and functioning may underlie the thermal acclimation of SOM decomposition and should be taken into account when predicting the response of soil CO2 efflux to global warming.

Growth Dynamics of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa as a Function of Time to Detection in BacT/Alert 3D Blood Culture Bottles with Various Preincubation Conditions

BackgroundDelayed entry of blood culture bottles is inevitable when microbiological laboratories do not operate for 24 hr. There are few studies reported for prestorage of these bottles. The growth dynamics of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were investigated with respect to various preincubation conditions.MethodsFifteen or 150 colony-forming units (CFU) of bacteria were inoculated into standard aerobic or anaerobic blood culture bottles. Bottles were preincubated at 25℃ or 37℃ for 0, 2, 4, 8, 12, 24, or 48 hr. The time to detection (TTD) then was monitored using the BacT/Alert 3D system (bioMerieux Inc., USA).ResultsSignificant difference in TTD was observed following preincubation for 8 hr at 25℃ vs. 4 hr at 37℃ for S. aureus, 4 hr at 25℃ vs. 4 hr at 37℃ for E. coli, 12 hr at 25℃ vs. 4 hr at 37℃ for P. aeruginosa, compared to no preincubation (P<0.005). TTD values did not vary significantly with bacterial CFU or with aerobic or anaerobic bottle type. The BacT/Alert 3D system returned false negatives following preincubation of P. aeruginosa for 48 hr at 25℃ or 24 hr at 37℃.ConclusionsTTD was mainly affected by preincubation temperature and duration rather than by input CFU quantity or bottle type for the 3 experimental bacteria.

Measuring basal soil respiration across Europe: Do incubation temperature and incubation period matter?

The European Commission recognises the essential role of soil biology in soil functioning and delivery of ecosystem services, but information is currently lacking evaluate of how these vary across soil and land-use types at a European scale. This study evaluated the measurement of the initial rate of soil basal respiration (BR) as a potential biological indicator of ecosystem service provision. The purpose of this study was to test ISO 16072:2002 (Soil Quality: Laboratory methods for the determining of microbial soil respiration). In the literature a range of pre-incubation temperatures (pre-inc) and experimental incubation temperatures (exp-inc) have been applied when using the ISO method for the establishment of basal respiration. This study evaluated whether the range of temperatures applied during pre- and exp- incubation had a significant effect on the rate of respiration determined when following the protocol established in ISO 16072:2002. The evaluation was carried out on a pedo-climatic gradient spanning ten countries across Europe and covering four biogeographical regions. Three sites were sampled in each country providing a range of soil and land-use parameters. Our results suggest that experimental incubation temperatures of 20°C or above should be used in the application of the methodology ISO 16072:2002 (incubation at 15°C resulted in erratic variation between replicates). However, pre-incubation temperature did not affect the soil basal respiration rate, when following the standard recommendations. The time interval with the best prediction of the initial rate of basal respiration was 6h.

Effects of a highly resistant rice starch and pre-incubation temperatures on the physicochemical properties of surimi gel from grass carp (Ctenopharyn Odon Idellus)

The effects of a specific rice starch (SRS), containing highly resistant starch (RSIII), on gel properties of grass carp (Ctenopharyngodon idella) and the influence of five levels of SRS (0%, 2%, 4%, 6%, and 8%w/w) on gel physicochemical properties at three different pre-incubation temperatures (4°C, 25°C, and 40°C) were investigated. Gels with increasing amounts of SRS addition showed lower expressible water contents and cooking loss values than did control gels. SDS gel electrophoresis revealed no changes in protein patterns, regardless of different SRS-added levels at the same pre-incubation temperature, but an evident decrease in the MHC when the pre-incubation temperature increased. The texture properties, colour attributes, SEM were optimal in the treatments containing 4%w/w SRS at the pre-incubation temperature 25°C. Thus, the optimum SRS addition level and pre-incubation temperature are proposed to be 4%w/w and 25°C, respectively.

Development and validation of automatic HS-SPME with a gas chromatography-ion trap/mass spectrometry method for analysis of volatiles in wines

An automated headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-ion trap/mass spectrometry (GC-IT/MS) was developed in order to quantify a large number of volatile compounds in wines such as alcohols, ester, norisoprenoids and terpenes. The procedures were optimized for SPME fiber selection, pre-incubation temperature and time, extraction temperature and time, and salt addition. A central composite experimental design was used in the optimization of the extraction conditions. The volatile compounds showed optimal extraction using a DVB/CAR/PDMS fiber, incubation of 5ml of wine with 2g NaCl at 45°C during 5min, and subsequent extraction of 30min at the same temperature. The method allowed the identification of 64 volatile compounds. Afterwards, the method was validated successfully for the most significant compounds and was applied to study the volatile composition of different white wines.

Curcumin's pre-incubation temperature affects its inhibitory potency toward amyloid fibrillation and fibril-induced cytotoxicity of lysozyme

BackgroundMore than twenty-seven human proteins can fold abnormally to form amyloid deposits associated with a number of degenerative diseases. The research reported here is aimed at exploring the connection between curcumin's thermostability and its inhibitory activity toward the amyloid fibrillation of hen egg-white lysozyme (HEWL). MethodsThT fluorescence spectroscopy, equilibrium thermal denaturation analysis, and transmission electron microscopy were employed for structural characterization. MTT reduction and flow cytometric analyses were used to examine cell viability. Results and conclusionThe addition of thermally pre-treated curcumin was found to attenuate the formation of HEWL fibrils and the observed fibrillation inhibition was dependent upon the pre-incubation temperature of curcumin. Our results also demonstrated that the cytotoxic effects of fibrillar HEWL species on PC 12 and SH-SY5Y cells were decreased and negatively correlated with curcumin's thermostability. Next, an enhanced stability of HEWL was perceived upon the addition of curcumin pre-incubated at lower temperature. Furthermore, we found that the alteration of curcumin's thermostability was associated with its inhibitory potency against HEWL fibrillation. General significanceWe believe that the results from this research may contribute to the development of effective therapeutics for amyloidoses.

Genetic studies of the role of fatty acid and coenzyme A in photocatalytic inactivation of Escherichia coli

The roles of bacterial cellular components, namely, fatty acid profile and coenzyme A, in photocatalytic inactivation of bacteria were investigated. Escherichia coli BW25113, as a “parental strain”, and its isogenic single-gene deletion mutants E. coli JW1081 (fabF− mutant) and E. coli JW3942 (coaA− mutant) showed different susceptibilities towards photocatalytic inactivation by titanium dioxide (TiO2, irradiated by UVA lamps (λ = 365 nm)). Regulating the fatty acid composition through pre-incubation temperature adjustment demonstrated the crucial role of cell membrane fatty acid profile in bacterial susceptibility towards photocatalytic inactivation, while the lower coenzyme A level in coaA− mutant correlated well with its lower susceptibility towards photocatalytic inactivation. Furthermore, transmission electron microscopic study demonstrated the photocatalytic destruction process of bacterial cells. This is the first study using single-gene deletion mutants to explore better understanding of the photocatalytic inactivation mechanism of E. coli.

Short-term temperature impact on soil heterotrophic respiration in limed agricultural soil samples

This study sought to investigate the hourly and daily timescale responses of soil CO2 fluxes to temperature in a limed agricultural soil. Observations from different incubation experiments were compared with the results of a model combining biotic (heterotrophic respiration) and abiotic (carbonate weathering) components. Several samples were pre-incubated for 8–9 days at three temperatures (5, 15 and 25 °C) and then submitted to short-term temperature (STT) cycles (where the temperature was increased from 5 to 35 °C in 10 °C stages, with each stage being 3 h long). During the temperature cycles (hourly timescale), the soil CO2 fluxes increased significantly with temperature under all pre-incubation temperature (PIT) treatments. A hysteresis effect and negative fluxes during cooling phases were also systematically observed. At a given hourly timescale temperature, there was a negative relationship of the CO2 fluxes with the PIT. Using the combined model allowed the experimental results to be clearly described, including the negative fluxes and the hysteresis effect, showing the potentially large contribution of abiotic fluxes to total fluxes in limed soils, after STT changes. The fairly good agreement between the measured and simulated flux results also suggested that the biotic flux temperature sensitivity was probably unaffected by timescale (hourly or daily) or PIT. The negative relationship of the CO2 fluxes with the PIT probably derived from very labile soil carbon depletion, as shown in the simulations. This was not, however, confirmed by soil carbon measurements, which leaves open the possibility of adaptation within the microbial community.

Upper lethal temperatures of Northern Bobwhite embryos and the thermal properties of their eggs

Northern Bobwhite eggs in the southern United States are often exposed to ambient temperatures in excess of their normal incubation temperature when unattended during their typical extended preincubation period. In drought years, typified by high ambient temperatures, Bobwhite eggs are often exposed to temperatures > 45°C, well-above the upper lethal temperature of most other birds. Because the upper lethal temperature of Bobwhite embryos is not currently known, simulated clutches of eggs were exposed to preincubation temperatures ranging from 39 to 52°C for exposure times of 1, 3, or 6 h. The upper lethal temperatures and the temperatures resulting in ≥ 50% death of Northern Bobwhite embryos were recorded in addition to the time to thermal equilibrium of Bobwhite eggs. The upper lethal temperature for 1, 3, and 6 h of preincubation exposure was 51, 49, and 46°C, respectively. The temperatures resulting in ≥ 50% death were 46, 44, and 40°C for eggs exposed to elevated temperatures for 1, 3, and 6 h, respectively. The mean time for the inner-egg temperature to reach the ambient temperature was 38 ± 1 min (± SE). The thermal tolerances of Northern Bobwhite embryos were much higher than expected, and among the highest reported for birds, indicating an adaptation to the naturally occurring temperature extremes that often occur in the Bobwhite's semi-arid southern range. However, as the temperature increased above the incubation temperature, hatching success declined, showing that increased thermal tolerance has a cost. Although Bobwhite producers, managers, and researchers will find this information useful, it seems most interesting that high temperatures could plausibly have contributed to the population decline observed in the Bobwhite's semi-arid range.