Suboptimal pH Research Articles

Digestive coordination of the gastric function in Atlantic salmon Salmo salar juveniles

Artificial diets have been reported to produce suboptimal pH values in the stomach of salmonid species. Therefore, it is interesting to investigate the gastric function of different species fed commercial diets. In the present work, two groups of Atlantic salmon juveniles were either fasted or fed with a commercial diet. The gastric and duodenal pH values were measured over a 25 h post-prandial period and, dry matter, moisture and protease activity in the gastric chyme of the fed group were also determined. In the fed group, the gastric pH dropped to 3.5, and duodenal pH increased from 7.0 to 7.5 at the 4th h post-feeding, whereas fasted fish showed no significant changes in the gastric pH or duodenal pH. Fed animals showed: i) two discrete events of evacuation, before the 2nd hand after the 8th h of digestion, ii) a change in the moisture of the gastric chyme from less than 10% in the diet to 60% at the 2nd h post-feeding, and iii) a maximal gastric proteases activity of 600 U mL -1 at the 2nd h followed by a nearly constant of value of ~300 U mL -1 . It can be concluded that the juveniles of the Atlantic salmon can suitably acidify a commercial diet to a value generally accepted as optimal for gastric proteases of fish, and to retain about 2/3 of the ingested diet under such conditions and at a nearly constant protease activity until the 8th h of digestion. Normal 0 21 false false false ES-CL X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:Tabla normal; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:Calibri,sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-fareast-language:EN-US;}

Improved freeze-dried kit for the preparation of 188ReN-DEDC/lipiodol for the therapy of unresectable hepatocellular carcinoma

Rhenium-188-N-(DEDC)2/lipiodol (abbreviated as 188ReN-DEDC, where DEDC = monoanionic diethyldithiocarbamate) is a clinically proven radiopharmaceutical for the therapy of unresectable hepatocellular carcinoma (HCC) through trans arterial radioembolization (TARE). A two-vial freeze-dried kit for the preparation of [188ReN(DEDC)2] complex using sodium perrhenate (Na188ReO4) obtained from a commercial Tungsten-188/Rhenium-188 generator had been reported earlier. This method required addition of stipulated volume of glacial acetic acid into vial 1 by the user for efficient preparation of [188ReN]2+ intermediate. An error in this step can result in low radiochemical yield of [188ReN]2+ intermediate as well as sub-optimal pH of the reaction mixture for the second step, leading to poor radiochemical purity of 188ReN-DEDC complex. In the present work, a solution to this problem was found by including an oxalate buffer of pH = 3 in vial 1, eliminating the need for the addition of glacial acetic acid by the user. This modification not only made the kits more user-friendly, it resulted in significant improvement in the kinetics of formation of [188ReN]2+ intermediate, wherein > 95% radiochemical purity could be achieved within 5 min incubation at ambient temperature. Moreover, the novel route for the preparation of [188ReN]2+ intermediate may be applied to any radiopharmaceutical based on 188ReN-core.

PH Change in Culture Medium Induces Reorganization of Intranuclear Actin in Two-Cell Mouse Embryos

Actin is a permanent component of the cell nucleus involved in many nuclear processes. However, some nuclear functions of actin remain insufficiently explored. The role played by various extracellular stimuli in regulation of nuclear actin still remains enigmatic. Deviation of basic parameters of culture medium from optimal values is a member of the group of extracellular stimuli that are very important for mammalian embryos cultured in vitro. Change in culture medium pH from the level optimal for embryo homeostasis is one such signals. The purpose of this study was to investigate the intranuclear actin distribution in nuclei of two-cell mouse embryos under stress conditions induced by changes in extracellular pH. The pattern of actin localization has been tracked after short-term culturing of the embryos at optimal (pH 7.2), increased (pH 7.8), or decreased (pH 6.5) pH conditions. Analysis was carried out with confocal microscopy using methods of direct fluorescent and indirect immunofluorescent identification of actin. It has been shown that the change of culture medium pH from the optimum value is the signal that alters intranuclear actin distribution in nuclei of the embryonic cells. Culture of two-cell mouse embryos in suboptimal pH conditions (pH 6.5 and pH 7.8) induced alterations in the intranuclear actin localization, which, in particular, were expressed in accumulation of monomeric actin and the appearance of phalloidin-stainable actin in the nuclei. These changes, in our opinion, show some signs of similarity with stress-induced changes in nuclear-actin distribution, which, as has been reported earlier by a number of researchers, have been observed in the nuclei of somatic cells.

Industrial robustness linked to the gluconolactonase from Zymomonas mobilis.

The physiological characteristics and the potential gluconolactone production of the gluconolactonase-deficient strain, Zymomonas mobilis ZM4 gnlΔ, were investigated via growth inhibitory assay and biotransformation of glucose and fructose into gluconolactone and sorbitol, respectively. The results of ethanol fermentation studies performed in the presence of high concentration of glucose (>200gl-1) under fermentative or aerobic conditions indicated that a significant reduction of volumetric ethanol productivity from the strain of ZM4 gnlΔ was noticeable due to the reduced rates of specific growth, sugar uptake, and biomass yield as compared with those of the parental strain ZM4. The biotransformation prepared at pH 6.0 using the permeabilized cell indicated that gluconic acid from ZM4 gnlΔ was still produced as a major product (67gl-1) together with sorbitol (65gl-1) rather than gluconolactone after 24h. Only small amount of gluconolactone was transiently overproduced up to 9g l-1, but at the end of biotransformation, all gluconolactone were oxidized into gluconic acid. This indicated that autolysis of gluconolactone at the pH led to such results despite under gluconolactonase inactivation conditions. The physiological characteristics of ZM4 gnlΔ was further investigated under various stress conditions, including suboptimal pH (3.5~6.0), temperature (25~40°C), and presence of growth inhibitory molecules including hydrogen peroxide, ethanol, acetic acid, furfural, and so forth. The results indicated that ZM4 gnlΔ was more susceptible at high glucose concentration, low pH of 3.5, and high temperature of 40°C and in the presence of 4mM H2O2 comparing with ZM4. Therefore, the results were evident that gluconolactonase in Z. mobilis contributed to industrial robustness and anti-stress regulation.

Sub-optimal pH Preadaptation Improves the Survival of Lactobacillus plantarum Strains and the Malic Acid Consumption in Wine-Like Medium.

Forty-two oenological strains of Lb. plantarum were assessed for their response to ethanol and pH values generally encountered in wines. Strains showed a higher variability in the survival when exposed to low pH (3.5 or 3.0) than when exposed to ethanol (10 or 14%). The study allowed to individuate the highest ethanol concentration (8%) and the lowest pH value (4.0) for the growth of strains, even if the maximum specific growth rate (μmax) resulted significantly reduced by these conditions. Two strains (GT1 and LT11) preadapted to 2% ethanol and cultured up to 14% of ethanol showed a higher growth than those non-preadapted when they were cultivated at 8% of ethanol. The evaluation of the same strains preadapted to low pH values (5.0 and 4.0) and then grown at pH 3.5 or 3.0 showed only for GT1 a sensitive μmax increment when it was cultivated in MRS at pH 3 after a preadaptation to pH 5.0. The survival of GT1 and LT11 was evaluated in Ringer's solution at 14% ethanol after a long-term adaptation in MRS with 2% ethanol or in MRS with 2% ethanol acidified at pH 5.0 (both conditions, BC). Analogously, the survival was evaluated at pH 3.5 after a long-term adaptation in MRS at pH 5.0 or in MRS BC. The impact of the physiologic state (exponential phase vs stationary phase) on the survival was also evaluated. Preadapted cells showed the same behavior of non-preadapted cells only when cultures were recovered in the stationary phase. Mathematical functions were individuated for the description of the survival of GT1 and LT11 in MRS at 14% ethanol or at pH 3.5. Finally, a synthetic wine (SW) was used to assess the behavior of Lb. plantarum GT1 and LT11 preadapted in MRS at 2% ethanol or at pH 5.0 or in BC. Only GT1 preadapted to pH 5.0 and collected in the stationary phase showed constant values of microbial counts after incubation for 15 days at 20°C. In addition, after 15 days the L-malic acid resulted completely degraded and the pH value increased of about 0.3 units.

Quantitative Detection of Oxidative Burst upon Activation of Plant Receptor Kinases.

The oxidative burst or the production of reactive oxygen species (ROS) is a typical cellular response of both plants and animals to diverse abiotic and biotic stresses. Mainly, the (re-)active oxygen species include the superoxide anion (O2-), hydrogen peroxide (H2O2), and the hydroxyl radical (OH•). Here, we outline the detection of extracellularly produced ROS in plant leaf pieces using a chemiluminescence-based bioassay with the luminol L-012 as a substrate being oxidized in the presence of ROS. Since this type of assay is in use in many laboratories, e.g., as a readout for activation of plant receptor kinases, we include a discussion on the interpretation of results and points addressing problems with the buffers at suboptimal pH values that negatively influence the chemiluminescence production.

Influence of food intrinsic factors on the inactivation efficacy of cold atmospheric plasma: Impact of osmotic stress, suboptimal pH and food structure

Abstract Cold atmospheric plasma (CAP) has proved to have great potential as a mild food decontamination technology. Different process parameters, including food intrinsic factors, are known to influence the resistance of the cells towards the treatment. The importance of osmotic stress (0, 2, 6% (w/v) NaCl) and suboptimal pH (5.5, 6.5, 7.4) on the CAP efficacy to inactivate Salmonella Typhimurium and Listeria monocytogenes is studied for various food structures. The helium-oxygen plasma was generated by a dielectric barrier discharge reactor, treating samples up to 10 min. If grown under osmotic stress or at suboptimal pH, microbial cells adapt and become more resistant during CAP treatment (stress hardening). Additionally, the microorganisms and the food structures also influence the inactivation results. This study illustrates the importance of increasing knowledge on food intrinsic factors, to be able to predict the final CAP inactivation result.

Influence of processing parameters and stress adaptation on the inactivation of Listeria monocytogenes by Non-Thermal Atmospheric Plasma (NTAP)

This study evaluated the effectiveness of Non-Thermal Atmospheric Plasma (NTAP) treatments against Listeria. Firstly, the impact of gas composition and flow rate on L. monocytogenes and L. innocua (used as a surrogate) inactivation by NTAP was monitored. Secondly, the influence of stress adaptation (growth under suboptimal conditions, using a wide range of temperatures and media acidified up to pH5.5 with citric, lactic, malic or hydrochloric acid, or short-term exposure to acid, cold or thermal shocks) on L. monocytogenes NTAP resistance was assessed. Survival curves obtained were concave upward. A mathematical model based on the Weibull distribution accurately described the inactivation kinetics. Both L. monocytogenes and L. innocua showed a higher sensitivity to plasma when the treatment was performed using air than when nitrogen was used. In fact, the use of nitrogen as working gas made the plasma treatment almost ineffective. The effect of gas flow rate on the effectiveness of the NTAP treatment depended on the type of gas used to generate plasma. Increases in flow rate from 5 to 10L/min caused an acceleration of bacterial inactivation when air was used, while an additional increase of gas flow from 10 to 15L/min had a minor impact on microbial inactivation. On the other hand, gas flow rate hardly affected NTAP treatment efficiency when nitrogen was used to generate plasma. L. monocytogenes growth under sub-optimal temperature or pH conditions or short-term exposure to acid, heat or cold stress conditions did not significantly modify its NTAP resistance. This suggests that temperature and pH stress adaptation does not induce a cross-protection response against NTAP treatments in L. monocytogenes, what makes NTAP an attractive technology for food decontamination within minimal processing strategies targeting this pathogenic microorganism.

PH and CO2 effects on Coelastrella (Scotiellopsis) rubescens growth and metabolism

We studied effects of рН and СО2 enrichment on the physiological condition and biochemical composition of a carotenogenic microalga Coelastrella (Scotiellopsis Vinatzer) rubescens Kaufnerova et Elias (Scenedesmaceae, Sphaeropleales, Chlorophyceae), a promising source of natural astaxanthin. The microalga was grown at a constant pH (5, 6, 7 or 8) maintained by direct СО2 injection. The air-sparged culture served as the control. Cell division rate and size, dry biomass productivity, the rates of nitrogen and phosphorus uptake as well as photosynthetic pigment and total lipid content and fatty acid composition were followed. С. rubescens possessed a narrow-range рН tolerance (the optimum рН 6–7). Under these conditions, the highest values of the maximum (1.0–1.1 1/day) and average (0.3–0.35 1/day) specific growth rate, chlorophyll а (4.8–4.9%) and total carotenoid dry weight percentages (1.7–1.8%) were recorded. Cell lipid fatty acid unsaturation index (1.851) and polyunsaturated fatty acid percentage (36–39%) and С18:3 ω3/С18:1 ω9 ratio (3.8–4.5) were also the highest under these conditions. A decline of рН to 5 brought about severe stress manifesting itself as a cell division cessation, photosynthetic apparatus reduction, two-fold increase in cell volume, accumulation of dry weight and lipids and a considerable decline in fatty acid unsaturation. Cultivation of С. rubescens without СО2 enrichment resulted in a rapid alkalization of the medium to рН 9.5–10.5 impairing the physiological condition of the cells. Reasons of the deteriorative effects of suboptimal pH values on the physiological condition of C. rubescens are discussed.

Influence of prebiotics on Lactobacillus reuteri death kinetics under sub-optimal temperatures and pH

Eaten foodstuffs are usually fortified with prebiotic ingredients, such as inulin and oligofructose (FOS). The main goal of this study was to evaluate the combined effects of inulin and FOS with either suboptimal pH or storage temperature on the viability of Lactobacillus reuteri DSM 20016. Data were modeled through Weibull equation for the evaluation of the microbiological shelf life and the survival time. Prebiotics enhanced the microbiological shelf life and enhanced the survival time of the target bacterium. The use of the factorial ANOVA highlighted that inulin and FOS exerted a different effect as a function of pH and temperature. Inulin prolonged survival time under acidic conditions, while the effect of glucose + FOS was significant at pH 8. Finally, temperature could act by increasing or decreasing the effect of prebiotics, as they could exert a protective effect at 30 °C but not at 44 °C. As the main output of this research, we could suggest that the effect of prebiotics on L. reuteri could be significantly affected by pH and temperature, thus pinpointing that the design of a symbiotic food should also rely on these factors.

Nutrient-Dependent Efficacy of the Antifungal Protein YvgO Correlates to Cellular Proliferation Rate in Candida albicans 3153A and Byssochlamys fulva H25.

YvgO is a recently characterized antifungal protein isolated from Bacillus thuringiensis SF361 that exhibits a broad spectrum of activity and pH stability. Customized colorimetric metabolic assays based on standard broth microdilution techniques were used to determine the variable tolerance of Byssochlamys fulva H25 and Candida albicans 3153A to YvgO exposure under select matrix conditions impacting cellular proliferation. Normalization of the solution pH after antifungal challenge expanded the available pH range under consideration allowing for a comprehensive in vitro assessment of YvgO efficacy. Indicator susceptibility was examined across an array of elementary growth-modifying conditions, including media pH, incubation temperature, ionic strength, and carbohydrate supplementation. Under suboptimal temperature and pH conditions, the indicator growth rate reduced, and YvgO-mediated susceptibility was attenuated. While YvgO association but not efficacy was somewhat influenced by solution ionic strength, carbohydrate supplementation was shown to be the most influential susceptibility factor, particularly for C. albicans. Although the specific choice of carbohydrate/nutrient supplement dictated the extent of enhanced YvgO efficacy, D-glucose additionally improved the association between antifungal and target. Indeed, when exposed to YvgO under conditions that lead to increased cellular proliferation, both indicators displayed a stronger association and susceptibility to YvgO when compared to carbohydrate-deprived media or suboptimal incubation environments. With further study, YvgO may have the capacity to function as a prophylaxis for food safety and preservation, as well as a pharmaceutical agent against opportunistic fungal pathogens either independently or in combination with other established treatments applied to both livestock and human health concerns.

Biofuel production: an odyssey from metabolic engineering to fermentation scale-up.

Metabolic engineering has developed microbial cell factories that can convert renewable carbon sources into biofuels. Current molecular biology tools can efficiently alter enzyme levels to redirect carbon fluxes toward biofuel production, but low product yield and titer in large bioreactors prevent the fulfillment of cheap biofuels. There are three major roadblocks preventing economical biofuel production. First, carbon fluxes from the substrate dissipate into a complex metabolic network. Besides the desired product, microbial hosts direct carbon flux to synthesize biomass, overflow metabolites, and heterologous enzymes. Second, microbial hosts need to oxidize a large portion of the substrate to generate both ATP and NAD(P)H to power biofuel synthesis. High cell maintenance, triggered by the metabolic burdens from genetic modifications, can significantly affect the ATP supply. Thereby, fermentation of advanced biofuels (such as biodiesel and hydrocarbons) often requires aerobic respiration to resolve the ATP shortage. Third, mass transfer limitations in large bioreactors create heterogeneous growth conditions and micro-environmental fluctuations (such as suboptimal O2 level and pH) that induce metabolic stresses and genetic instability. To overcome these limitations, fermentation engineering should merge with systems metabolic engineering. Modern fermentation engineers need to adopt new metabolic flux analysis tools that integrate kinetics, hydrodynamics, and 13C-proteomics, to reveal the dynamic physiologies of the microbial host under large bioreactor conditions. Based on metabolic analyses, fermentation engineers may employ rational pathway modifications, synthetic biology circuits, and bioreactor control algorithms to optimize large-scale biofuel production.

Sustainable nutrient management at field, farm and regional level: Soil testing, nutrient budgets and the trade-off between lime application and greenhouse gas emissions

Pollution from agriculture has environmental consequences at local and global scales. Managing this pollution is challenging because of diffuse sources and complex relationships between aquatic and atmospheric emissions. We illustrate this for a UK county that has suffered outbreaks of microbial pollution and eutrophication. We surveyed 49 livestock farms covering 12% of total agricultural grazed land. Soil nutrient status and whole-farm nutrient balances were determined, and the environmental impact of alleviating sub-optimal soil pH by liming was estimated at the county level. Only 37% of fields contained more P than was required for satisfactory grass growth, and soil acidity and available K were often limiting production. The mean farm N, P and K balances were similar to a modelled farm in England & Wales and EU indicators for the majority of North West Europe. This suggests that local eutrophication events linked to agriculture are more likely to relate to improper timing of nutrient application rather than over-application. None of the surveyed farmers used nutrient decision support tools, largely due to a lack of awareness and competing sources of information. Liming soils to pH 6.0 was estimated to both reduce N-leaching and N2O emissions; however, the net climate-change impact would be negative as the direct CO2 emissions would exceed CO2 equivalent emissions of not liming by 394% (95% CI 201–21,232). Although liming currently presents a net cost to farmers, a sensitivity analysis suggests that reduced lime cost could lead to economic benefit to farmers but still increased greenhouse gas emissions. The results are applicable to all pasture-based agricultural systems where there is a drive to maintain or increase production through optimal soil and nutrient management. The findings demonstrate an important trade-off between reducing aquatic and atmospheric pollution and agricultural productivity, and the need to improve communication of this trade-off to farmers.

Short Communication: Ruminal pH profile and feeding behaviour of feedlot cattle transitioning from a high-forage to a high-concentrate diet

Holtshausen, L., Schwartzkopf-Genswein, K. S. and Beauchemin, K. A. 2013. Short Communication: Ruminal pH profile and feeding behaviour of feedlot cattle transitioning from a high-forage to a high-concentrate diet. Can. J. Anim. Sci. 93: 529–533. Sixteen ruminally cannulated cattle (384±33.9 kg) were transitioned from a high-forage to a finishing diet over 21 d using three steps (+15% concentrate DM wk−1). Following the initiation of each transition step, mean daily ruminal pH declined and DM intake, meal size and meal duration were reduced. Recovery time to baseline feeding behaviour increased with severity of acidosis. Changes in feeding behaviour can be indicative of sub-optimal ruminal pH conditions during dietary transition. Transition protocols such as more steps, smaller grain increments or shorter initial and longer final steps may aid in reducing the incidence of acidosis.

P53 is involved in shrimp survival via its regulation roles on MnSOD and GPx in response to acute environmental stresses

The tumor suppressor gene p53 plays a critical role in safeguarding the integrity of genome in mammalian cells. It acts as a sequence-specific transcription factor. Once activated by a variety of cellular stresses, p53 transactivates downstream target genes, through which it regulates cell cycle and apoptosis. However, little is known about p53 as well as its downstream target genes in invertebrates. A full length cDNA that encodes a 453-amino-acid p53 protein (Lvp53) was characterized in the Pacific white shrimp (Litopenaeus vannamei) to explore the potential relationships between p53 and two antioxidant enzyme genes: Mn-superoxide dismutase (MnSOD) and glutathione peroxidase (GPx) in eliminating cell stresses in L. vannamei. Sequence analysis revealed a close phylogenetic relationship between Lvp53 and that of Marsupenaeus japonicus, and a high degree of conservation in critical amino acids residues is involved in DNA and zinc binding among species. Quantitative real-time PCR showed that Lvp53 was expressed with varied levels in all the 11 tissues under investigation. In response to acute pH challenge, the relative expression of Lvp53 was induced in a pH- and time-dependent manner, with the peak observed at pH6.1 and after 24h of treatment, in which condition, both the relative mRNA expressions and the enzymatic activities of LvMnSOD and LvGPx were increased correspondingly. In response to acute cadmium (Cd) exposure, the relative expression of Lvp53 was upregulated in a time- and concentration-dependent manner, with the maximum detected at Cd 6.6μM and after 48h of exposure, in which case, both the transcripts and the enzymatic activities of LvMnSOD and LvGPx were also induced. After Lvp53 transcripts were declined by double-strand RNA injection, the relative mRNA expressions of LvMnSOD and LvGPx were decreased correspondingly. Meanwhile, pH6.1 or 6.6μM Cd could not induce the transcripts or the enzymatic activities of LvMnSOD or LvGPx any more in Lvp53-silenced shrimp, but increased shrimp mortalities. These results indicated the involvement of Lvp53, LvMnSOD and LvGPx in mediating cell stress caused by suboptimal pH and elevated levels of Cd in L. vannamei, and that the expressions of LvMnSOD and LvGPx were positively regulated by Lvp53, which is a potential mechanism for shrimp to survive the oxidative stress that occurs during short-term exposure to Cd or challenge with acidic pH. This finding will contribute to better understanding of p53 signaling pathways and redox regulation in invertebrate organisms.

Electrophysiological Investigation of the Na+/H+ Antiporter NHAA from E. Coli.

NhaA from E. coli serves as a prototype for Na+/H+ antiporters. It was proposed that the pH dependent change of the transport activity is due to the transition of the transporter from an inactive to an active conformation, which is allosterically controlled by a pH sensor.NhaA is electrogenic. Therefore it is possible to perform electrophysiological measurements with NhaA using the solid supported membrane based electrophysiology (Mager et al., 2011). The SSM based electrophysiology permits measurements with a high time resolution and defined conditions on both sides of the membrane.The kinetic analysis of the SSM measurements shows that the decrease of the transport activity at suboptimal pH values is due to the competitive binding of the substrates to a common binding site (Mager, et al., 2011). Based on this finding we employed a minimal kinetic model. It explains the substrate dependence of the transport activity over a large concentration range.The changed pH dependencies of the mutants V254C NhaA and H225R NhaA were explained with the impairment of the allosteric mechanism of inactivation (Padan, 2008). However the phenomenological alterations of the transport properties can be explained by our minimal kinetic model, which does not include an allosteric mechanism of pH regulation. The altered transport properties are the result of a change of the substrate binding affinities or the rates of the conformational transitions.1. Padan, E., Trends Biochem Sci 33, 435-443 (2008).2. Mager, T. et al., J Biol Chem 286, 23570-23581 (2011).

Analysis of acidity production during enhanced reductive dechlorination using a simplified reactive transport model

Build-up of fermentation products and hydrochloric acid at a contaminated site undergoing enhanced reductive dechlorination can result in groundwater acidification. Sub-optimal pH conditions can inhibit microbial activity and lead to reduced dechlorination rates. The extent of acidification likely to occur is site-specific and depends primarily on the extent of fermentation and dechlorination, the geochemical composition of soil and groundwater, and the pH-sensitivity of the active microbial populations. Here, the key chemical and physical mechanisms that control the extent of groundwater acidification in a contaminated site were examined, and the extent to which the remediation efficiency was affected by variations in groundwater pH was evaluated using a simplified process-based reactive-transport model. This model was applied successfully to a well-documented field site and was then employed in a sensitivity analysis to identify the processes likely to significantly influence acidity production and subsequent microbial inhibition. The accumulation of organic acids produced from the fermentation of the injected substrate was the main cause of the pH change. The concentration of dissolved sulphates controlled substrate utilisation efficiency because sulphate-reducing biomass competed with halo-respiring biomass for the fermentation products. It was shown further that increased groundwater velocity increases dilution and reduces the accumulation of acidic products. As a consequence, the flow rate corresponding to the highest remediation efficiency depends on the fermentation and dechlorination rates. The model enables investigation and forecasting of the extent and areal distribution of pH change, providing a means to optimise the application of reductive dechlorination for site remediation.

Differential expression of particulate methane monooxygenase genes in the verrucomicrobial methanotroph ‘Methylacidiphilum kamchatkense’ Kam1

Methane monooxygenases (MMOs) are oxygen-dependent enzymes that catalyze the oxidation of methane to methanol in the methanotrophic bacteria. The thermoacidophilic verrucomicrobial methanotroph 'Methylacidiphilum kamchatkense' Kam1 contains three complete and phylogenetically distinct copies of the pmoCAB gene cluster apparently organized as operons, each encoding all three subunits of particulate MMO (pMMO), and a truncated pmoCA cluster encoding only two of the subunits. Two of the clusters are present as a tandem array, but the other clusters occur in isolation. Here, the expression of these clusters has been assessed using the four pmoA genes as targets in reverse transcriptase quantitative PCR analysis. One of the pmoA genes, designated pmoA2, is at least 35-fold more strongly transcribed than the other pmoA copies. Growth at suboptimal temperature and pH conditions did not significantly change the transcription pattern, indicating that the pmoCAB2 cluster encodes the functional pMMO under methane-fuelled growth conditions. During growth on methanol, expression of pmoA2 was reduced approximately tenfold as compared to growth on methane, suggesting a role for the alternative carbon substrates in gene regulation.

Correlation between bio-hydrogen production and polyhydroxybutyrate (PHB) synthesis by Rhodopseudomonas palustris WP3-5

The aim of this study was to determine the competition between H2 production and polyhydroxybutyrate (PHB) accumulation of Rhodopseudomonas palustris WP3-5 when grown on six different substrates. From the results, strain WP3-5 can utilize acetate, propionate, malate, and lactate to produce H2 but can only synthesize PHB on acetate and propionate. The substrate conversion efficiency (SCE) on acetate and propionate increased significantly after the maximum PHB content was achieved, illustrating a competition for reducing power when PHB synthesis occurred. However, when strain WP3-5 was cultivated at suboptimal pH values on acetate, the synthesized PHB prevented strain WP3-5 from the stress of the inappropriate pH and retained H2 producing efficiency as at optimal pH value. Consequently, although PHB synthesis does compete with H2 production in R. palustris WP3-5, it is still conducive to H2 production when strain WP3-5 is in a stressful condition.

Single-Cell Growth Probability of Listeria monocytogenes at Suboptimal Temperature, pH, and Water Activity

The single-cell growth probability of Listeria monocytogenes was characterized in tryptone soy broth supplemented with 0.6% yeast extract at temperature ranging from 5 to 25°C, pH ranging from 4.4 to 6.5, and water activity ranging from 0.919 to 0.989. Growth was monitored visually in 96-microwell plates and the growth probability was deduced from concentrations estimated with the MPN calculation. Models were proposed to describe the increase of the probability from 0 to 1 with increasing values of environmental factors. An exponential model was used for the temperature and the individual minimal temperature for growth ranged from −3.6°C [−4.5°C, −2.7°C] to 17.3°C [16.0°C, 18.6°C]. An inverse exponential model was convenient for the pH and the minimal pH for growth of individual cells ranged from 4.34 [3.93, 4.75] to 5.93 [4.85, 7.01]. A linear model was used for the water activity and the minimal value for growth of individual cells ranged from 0.917 [0.909, 0.925] to 0.988 [0.966, 1]. In spite of the great between-experiment variability, the minimal values estimated for the growth limits of individual cells were in accordance with values usually proposed for L. monocytogenes populations. This study provides models allowing the prediction of the growth probability of a few cells contaminating chilled foods with suboptimal pH and water activity improving thus the assessment of the behavior of L. monocytogenes cells naturally contaminating foods.